RE-REVIEW Supplement Book 1 Alkyl Esters CIR EXPERT PANEL

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RE-REVIEW Supplement Book 1

CIR EXPERT PANEL MEETING MARCH 5-6, 2012 ALKYL ESTERS RE-REVIEW – SUPPLEMENTAL BOOK 1 Cetyl Esters and Other Previously Reviewed Alkyl Esters

Cetyl Esters ...... 1 Isopropyl Linoleate ...... 9 Cetearyl Octanoate (now Cetearyl Ethylhexanoate) ...... 15 Octyl, Cetyl, and Isopropyl Palmitate ...... 25 Decyl and Isodecyl Oleate ...... 48 Butyl, Cetyl Isobutyl, Isocetyl, Isopropyl, Myristyl, and Octyl Stearate ...... 59 Isostearyl Neopentanoate ...... 99 Arachidyl Propionate ...... 121 Isopropyl Isostearate ...... 131 Cetyl, Isopropyl, and Octyldodecyl Ricinoleate ...... 138 Myristates ...... 186 Isononanoates and Nonanoates ...... 211 Stearyl Heptanoate, Caprylate, Palmitate, Stearate, Behenate, and Olivate ...... 288 Decyl, Ethylhexyl, Isodecyl, Lauryl, Octyldodecyl, and Tridecyl Cocoate ...... 299

Alkyl Esters Supplement Book 1

FINAL REPORT ON THE SAFETY ASSESSMENT OF CETYL ESTERS

Cetyl Esters is a synthetic wax composed of a mixture of esters of saturated fatty acids and fatty alcohols with carbon chain lengths between 14 and 18. Cetyl Esters is currently used as a skin conditioning agent-emollient in over 200 cosmetic formulations at concentrations of up to 7%. The esters that are found in Cetyl Esters include Cetyl Palmitate, Cetyl Stearate, Myristyl Myristate, Myristyl Stearate, Cetyl Myristate, and Stearyl Stearate. Safety data on four of these (Cetyl Palmitate, Cetyl Stearate, Myristyl Myristate, and Myristyl Stearate) have previously been reviewed and are summarized in this report; overall, the data show no systemic toxicity, no sensitization or photosensitization, and little irritation. Because of the structural similarity of Cetyl Myristate and Stearyl Stearate to these four ingredients, it is expected that they would behave in a similarly nontoxic manner. Therefore, it was concluded that Cetyl Esters (as a mixture of all six ingredients) is safe as used in cosmetics.

The following is a compilation of data concerning Cetyl Esters. Cetyl Esters is a synthetic mixture consisting of esters of saturated fatty acids and fatty alcohols; the fatty acids and alcohols have carbon chain lengths between 14 and 18. A synonym for Cetyl Esters is synthetic spermaceti wax (National Formulary, 1990; Nikitakis and McEwen, 1990; Wenninger and McEwen, 1995a). Within the National Formulary definition, there are nine esters that fit the specifications. One industry source reports that of these nine, three (Myristyl Palmitate, Stearyl Palmitate, and Stearyl Myristate) are not found in any commercial Cetyl Esters preparation. The source reports Cetyl Esters to be comprised of a mixture of one or more of the following six esters: Cetyl Palmitate, Myristyl Myristate, Cetyl Stearate, Myristyl Stearate, Cetyl Myristate, and Stearyl Stearate (Cosmetic, Toiletry and Fragrance Association [CTFAI, 1995). Of these six, the first four have been reviewed by the Cosmetic Ingredient

‘Reviewed by the Cosmetic Ingredient Review Expert Panel. Bindu Nair, Scientific Analyst and Writer, prepared this report. Address correspondence to Dr. F. Alan Andersen, Cosmetic Ingredient Review, 1101 17th Street NW, Suite 310, Washington, DC 20036, USA.

International Journal of ‘Ibxicology, 16 (Suppl. 1):122-HO,1997 123 Copyright 0 1997 Cosmetic Ingredient Review 1091-Fi818/97$12.99 + .ocl

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Review (CIR) Expert Panel and found to be safe as used (Elder 1982a, 1982b, 1985). The expected concentration of use in cosmetics of these four are Cetyl Palmitate (lo%), Myristyl Myristate (25%), Cetyl Stearate (lo%), and Myristyl Stearate (5%). Following a Notice to Proceed (without preparing a Scientific Literature Review), the CIR Expert Panel decided to continue its review of Cetyl Esters, with use of the findings of the earlier reports. Adaptions from these earlier reviews appear in this report in an itali- cized font. In addition, because these reviews were completed more than 10 years ago, an updated computer literature search was performed. No studies relevant to the safety assessment have been published since the CIR review.

DEFINITION AND STRUCTURE Cetyl Esters is a synthetic wax with a composition and properties identical to natural spermaceti wax (Wenninger and McEwen, 1995a). The structure of the six esters that can be present in a Cetyl Esters preparations are shown in Figure 1.

PHYSICAL AND CHEMICAL PROPERTIES Cetyl Esters is described as a somewhat translucent white to off-white solid with a crystalline structure. It has a faint odor and a bland mild

P H CH@l.$,,C -OCH,GH,)14CHj CHS(CHZ),SC-OCHZlCH~)14CH3

CETYL PALMITATE CETYL STEARATE

P P CH3(CH.&C -0CH,fCH2,1#H3 CH3(CH$&OCH2(CH2)12CH3

MYRISTYL MYRISTATE MYRISTYL STEARATE

& P CH3PZH2),4C -OCH2(CH2),4CH3 CH3(CHZ),gC-OCHZlCH~)16CH3

CENL MYRISTATE’ STEARYL STEARATE’

Figure 1. Structure of the six esters found in Cetyl Esters. *Not previously reviewed by CIR.

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CETYL ESTERS 125 taste. It is soluble in boiling alcohol, ether, chloroform, and fixed oils and is insoluble in water and cold alcohol. It has a specific gravity range of 0.820 to 0.840 at 5O”C, a melting range between 43” and 47°C and a saponification value of 109 to 120 (Nikitakis and McEwen, 1990).

USE Cosmetic Cetyl Esters is used in cosmetic formulations as a skin-conditioning agent-emollient (Wenninger and McEwen, 199513). As of January 1995, there were 210 reported uses of Cetyl Esters in various cosmetics (Food and Drug Administration [FDA], 1995; Table 1). Concentrations of use are no longer reported to the FDA (FDA, January 1992). Data submitted directly to CIR from the cosmetics industry indicate that Cetyl Esters is used at concentrations of up to 7% in both leave-on and rinse-off cosmetic formulations (CTFA, 1995).

GENERAL BIOLOGY The Myristates, Palmitates, and Stearates can undergo chemical or enzymatic hydrolysis to myristic acid, palmitic acid, and stearic acid (and the corresponding alcohols), respectively. Tkansesterification and other typical ester reactions may also occur. All of these esters are saturated compounds and would not be expected to autoxidize readily. Myristic acid is a digestible constituent of most vegetables and animals and is nontoxic when ingested. When myristic acid was fed to dogs (as the ethyl ester), less than 2% of the amount fed was recovered as unabsorbed material in the feces; there was no increase in ether-soluble acids in the urine. The palmitates would be expected to be nontoxic in view of their hydrolysis to palmitic acid (Elder, 1982a, 19823, 1985).

ANIMAL TOXICOLOGY Acute Toxicity Oral The oral LD,, in CFW mice of the Carworth strain of a trade product composed of 60% to 65% of Cetyl Esters in corn oil was more than 20 g/kg (Leberco Laboratories, 1971a). Another source reported the oral LD,, in hairless mice for Cetyl Palmitate was more than 5000 mg/kg (Henkel Department of Toxicology, 1974). Myristyl Myristate, undiluted and diluted with corn oil or propylene glycol, was nontoxic when fed to rats. The acute oral LD,, for Myristyl Myristate was more than 14.4 g/kg in rats; for Cetyl Palmitate, more

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Table 1. Frequency of use of Cetyl Esters

No. of formulations No. containing Product category in category ingredient

Eyebrow pencil 100 1 Eye shadow 597 3 Mascara 211 1 Other eye makeup preparation 130 4 Hair conditioner 693 8 Hair straighteners 59 1 Other hair preparations 382 2 Hair dyes and colors (all types) 1497 15 Blushers (all types) 283 2 Foundations 333 25 Lipstick 997 26 Makeup bases 159 7 Makeup fixatives 11 4 Other makeup preparations 155 11 Nail creams and lotions 18 1 Bath soaps and detergents 339 2 Deodorants (underarm) 293 5 Other personal cleanliness 317 2 Other shaving preparations 60 1 Cleansing 771 10 Face and neck 261 6 Bodyandhand 987 21 Moisturizing 873 31 Night 220 5 Paste masks (mud packs) 276 4 Other skin care preparations 782 6 Suntan gels, creams, liquids 196 4 Other suntan preparations 62 2 1995 Totals 210

Source. FDA, 1995.

than 14.4 g/kg in rats; and for Myristyl Stearate, more than 10 glkg in mice, when diluted in corn oil (Elder, 1982a, 19823, 1985).

Dermal Myristyl Myristate, in studies using rabbits, had no dermal toxicity (dose 2.0 gRzg; Elder, 1982a). A 50% slurry of Cetyl Palmitate in distilled water was not dermally toxic to rabbits when applied in a single 24-h occlusive patch at a dose of 2.OgLkg. Slight irritation was noted (Elder, 1982b).

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CETYL ESTERS 127

Irritation Dermal A trade product containing 60% to 65% Cetyl Esters was applied in a 0.5-g dose to intact and abraded skin of three albino rabbits. The sites were covered for 24 h. Scoring was done at the time of patch removal and 48 h later. The primary irritation index (PII) was 0 (Leberco Laboratories, 1971b). I A 50% dilution in oil (type unspecified) of Cetyl Palmitate applied twice daily for 5 days to five hairless mice was nonirritating (Henkel Department of Toxicology, 1974). Similarly, a 50% dilution applied in a 24-h patch exposure was nonirritating to rabbits (number not reported; Henkel Department of Toxicology, 1974). Ocular The right conjunctival sac of each of three albino rabbits was instilled with 0.1 g of a trade product containing 60% to 65% Cetyl Esters. The left eye(s) served as control. Examinations were performed every 24 h for 4 days, and then at day 7. No irritation was observed (Leberco Laboratories, 1971c). In seven studies, Myristyl Myristate caused minimal to mild skin irritation and minimal eye irritation (Elder, 1982a). Four separate studies tested the irritancy of Cetyl Palmitate using the Draize primary skin irritation technique. Occlusive patches were applied for 24 h. Reactions were evaluated at 24 h and 72 h readings and the PII scores were calculated as an average of the two readings. The maximum possible PII score was 8. The PIIs were as follows: 0.5 mL of a 570 w/w dispersion, score 0.38 (0.0 at 72 h); 0.5-g effective dosage, score 0; 0.5g, score 0.17; and 0.5 mL, score 0.4 (0.0 at 72 h). ltvo additional irritation studies tested formulations containing 2.5% and 2.7% Cetyl Palmitate and had PIIs of 1.0 and 0.9, respectively. In four studies using the Draize eye irritation technique, Cetyl Palmitate was minimally irritating (Elder, 1982b). Undiluted Myristyl Stearate had PII scores of 0.0 in rabbits after a 24-h occlusive patch exposure. Undiluted Myristyl Stearate caused slight, transient ocular irritation in rabbits (24-h Draize scores of 2, 4, and 6 out of a maximum of 110; no irritation was observed at later evaluations; Elder, 1985).

Sensitization In guinea pig sensitization studies, Myristyl Myristate produced no evidence of skin sensitization (Elder, 1982a). A 1% suspension of Cetyl Palmitate was minimally irritating but did not appear to be a sensitizing agent in guinea pigs (Elder, 1982b).

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CLINICAL ASSESSMENT OF SAFETY Five volunteers were patch-tested with a 50% dilution of Cetyl Palmitate in oil (type unspecified). Cetyl Palmitate was considered a nonirritant (Henkel Department of Toxicology, 1974). Details were not reported. A formulation containing 8% Myristyl Myristate was minimally irritating in a human primary skin irritation study. The same product produced no evidence of sensitization in a human skin sensitization study (Elder, 1982a). Various clinical studies were performed using trade products containing 2.5% or 2.7% Cetyl Palmitate. There was no evidence of primary irritation. Cetyl Palmitate was considered to be a <‘weak potential sensitizer.” There were no cases of phototoxicity when the 2.7% formulation was tested using 10 normal subjects, and it was considered to have ‘low potential for photo-contact allergenicity” when tested using 25 normal subjects. A safety-in-use study tested the 2.5% formulation using 30 women; no irritation was noted. In a larger-scale study using 150 women and the 2.7% formulation, it was concluded that the formulation had a low potential for irritation (Elder, 1982b). Results of clinical studies indicate that the Stearates and cosmetic products containing Stearates are essentially nonsensitizing; nonphototoxic; nonphotosensitizing; and, at most, minimal or mild skin irritants. The highest concentrations of Cetyl Stearate and Myristyl Stearate tested in the various studies were 50% and 9.8%, respectively (Elder, 1985). (Note: Photosensitization and phototoxicity studies were not conducted on these two stearates.)

SUMMARY The mixture Cetyl Esters is a synthetic wax composed of a combination of the following six esters: Cetyl Palmitate, Cetyl Stearate, Myristyl Myristate, Myristyl Stearate, Cetyl Myristate, and Stearyl Stearate. The CIR Expert Panel had previously conducted safety assessments on the first four ingredients. Cetyl Esters is used in cosmetic formulations as a skin-conditioning agent-emollient. The oral LD,, (mice) of a product containing 60% to 65% Cetyl Esters was more than 20 g/kg; for Myristyl Myristate, more than 14.4 g/kg (rats); for Cetyl Palmitate, more than 5 g/kg (rats); and for Myristyl Stearate, more than 10 g/kg (mice). In acute dermal studies, 2.0 g/kg Myristyl Myristate was nontoxic to rabbits. Slight irritation was noted when 2.0 g/kg Cetyl Palmitate was applied in a 24-h patch.

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CETYL ESTERS 129

No irritation was noted with 0.5 g of a formulation containing 60% to 65% Cetyl Esters was applied to intact and abraded rabbit skin. The same formulation was not an ocular irritant for rabbit eyes, Myristyl Myristate caused mild skin irritation and minimal eye irritation. Cetyl Palmitate was not a dermal irritant and was minimally irritating for rabbit eyes. Myristyl Stearate was nonirritating to rabbit skin after a 24-h occlusive patch exposure, and was a slight, transient ocular irritant. No evidence of sensitization was found when Myristyl Myristate was tested; Cetyl Palmitate caused minimal irritation but no sensitization. In clinical dermal studies, 50% Cetyl Palmitate was found to be nonirritating. Other studies found a formulation containing 2.7% Cetyl Palmitate to be a weak potential sensitizer and a low potential irritant. There were no instances of phototoxicity. A formulation containing 8% Myristyl Myristate was minimally irritating but not sensitizing. Stearates of varying concentrations were essentially nonsensitizing, nonphototoxic, and nonphotosensitizing, and were minimal to mild skin irritants.

DISCUSSION The CIR Expert Panel used data from four previously reviewed esters of fatty acids and fatty alcohols, and their structural similarity to the Cetyl Esters is evidence to suggest that the Cetyl Esters mixture is safe as used in cosmetics.

CONCLUSION Based on the available data, the CIR Expert Panel concludes that Cetyl Esters is safe as used in cosmetics.

REFERENCES Cosmetic, Toiletry and Fragrance Association (CTFA). 1995. Use Levels for Various Ingredients. Unpublished data submitted by CTFA.* Elder R., ed. 1982a. Final report on the safety assessment of myristyl myristate and Isopropyl myristate. J. Am. Coil. Toxicol. 1(4):55-80. Elder R., ed. 1982b. Final report on the safety assessment of octyl palmitate, cetyl palmitate and isopropyl palmitate. J. Am. CM. Toxicol. 1(2):X3-35. Elder R., ed. 1985. Final report on the safety assessment of butyl stearate, cetyl stearate, isobutyl stearate, isocetyl stearate, isopropyl stearate, myristyl stearate, and octyl stearate. J. Am. CoZZ. Toxicol. 4(5):107-146.

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Food and Drug Administration (FDA). 1992. Modification in Voluntary Filing of Cosmetic Product Ingredient and Cosmetic Raw Composition Statements. Federal Register 57:3123-3130. FDA. 1995. Frequency of use of cosmetic ingredients. FDA database. Washington, DC: FDA. Henkel Department Of Toxicology. 1974. Cutina CP/Cetyl Palmitate/ Toxicological Data. Report No. 740038. Unpublished data submitted by Henkel KgaA.* Leberco Laboratories. 1971a. LD,, of Cetina No. 718 in mice. Assay #: 16865. Unpublished data submitted by CTFA.* Leberco Laboratories. 1971b. Primary irritation of Cetina No. 718 in mice. Assay #: 16866. Unpublished data submitted by CTFA.* Leberco Laboratories. 1971c. Ocular irritation of Cetina No. 718 in mice. Assay #: 16867. Unpublished data submitted by CTFA.* National Formulary. 1990. The United States Pharmacopeia. Rockville, MD: US Pharmacopeial Convention. Nikitakis J. M., and McEwen G. N., Jr. eds. 1990. CTFA Compendium of Cosmetic Ingredient Composition-Descriptions I. Washington, DC: CTFA. Wenninger J. A, and McEwen G. N., Jr. eds. 1995a. International Cosmetic Ingredient Dictionary, 6th ed. Washington, DC: CTFA. Wenninger J. A, and McEwen G. N., Jr. eds. 1995b. International Cosmetic Ingredient Handbook, 3rd ed. Washington, DC: CTFA.

*Available for review: Director, Cosmetic Ingrediant Review, 1101 17th Street, NW, Washington, DC 20036.

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Final Report on the Safety Assessment of Isopropyl Linoleate

Isopropyl Linoleate is the ester of isopropyl alcohol and linoleic acid. In cosmetics, it is used as a skin conditioning agent and emollient at concentrations ranging from 0.1% to 10.0%. In an acute oral toxicity study, none of the albino rabbits that received doses of 10.0% Isopropyl Linoleate in corn oil died. Isopropyl Linoleate (undiluted and 10.0% suspension) were classified as slight ocular irritants. Undiluted Isopropyl Linoleate was classified as a slight skin irritant. The report concludes that the safety of use of Isopropyl Linoleate has not been documented and substantiated, and that it is not possible to conclude that the ingredient is safe for use in cosmetic products. The report details the type of safety test data that is needed to substantiate the safety of use of Isopropyl Linoleate in cosmetic products.

INTRODUCTION

ata available to the Cosmetic Ingredient Review (CIR) concerning the chemistry, D use in cosmetic products, acute oral toxicity, and the skin and ocular irritation potential of Isopropyl Linoleate are included in this report. The types of data that are necessary for completion of the safety assessment of Isopropyl Linoleate are listed in the Discussion section.

CHEMISTRY

Chemical and Physical Properties

The commercial product generally known as Isopropyl Linoleate is a mixture consisting of: isopropyl linoleate (64.0~82.0%), isopropyl myristate (0.6% maximum), isopropyl palmitate (2.0-9.0%), isopropyl stearate (3.0% maximum), isopropyl oleate (9.0 to 25.0%), isopropyl linoleate (7.0% maximum), and free linoleic acid (1.75% maximum).“,*’ 51

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The main component isopropyl linoleate is the ester of isopropyl alcohol and linoleic acid that conforms generally to the formula:‘3’ CH3(CHd41H CH I w I

i” 51 CH(CH,),C-OCH(CHJ,

The ester is also known as: Linoleic Acid, Isopropyl Ester; 1-Methylethyl-9,12 Octadecadienoate; and 9,12-Otadecadienoic Acid, 1 -Methyl Ethyl Ester. It is a pale yellow, oily liquid consisting of fatty acids derived from a vegetable oil. The results of an ultraviolet (UV) spectral analysis of Isopropyl Linoleate (50 ppm in isopropyl alcohol) did not indicate absorbance in the UVB band (h max = 206 nm).‘4’ Properties of Isopropyl Linoleate are summarized in Table 1.

Analytical Methods

Isopropyl Linoleate has been analyzed via infrared spectroscopy and gas chromatography.‘*,”

COSMETIC USE

Isopropyl Linoleate is used as a skin conditioning agent-emollient in cosmetics.‘6) The FDA cosmetic product formulation computer printout”’ is compiled through voluntary filing of such data in accordance with Title 21 part 720.4 of the Code of Federal Regulations. (K Ingredients are listed in preset concentration ranges under specific product type categories. Since certain cosmetic ingredients are supplied by the manufacturer at less than 100% concentration, the value reported by the cosmetic formulator may not necessarily reflect the actual concentration found in the finished product; the actual concentration would be a fraction of that reported to the FDA. Data submitted within the framework of preset concentration ranges provide the opportunity for overestimation of the actual concentration of an ingredient in a particular product. An entry at the lowest end of a concentration range is the same as one entered at the highest end ofthat range, thus introducing the possibility of a two- to ten-fold error in the

TABLE 1. PROPERTIES OF ISOPROPYLLINOLEATE”,”

Form Pale yellow, oily liquid Solubility Soluble in mineral oil and vegetable oil, partly soluble in alcohol, insoluble in water Specific gravity at 25°C 0.8600 to 0.8700 Refractive index at 25°C 1.453 to 1.454 Acid value 3.5 maximum Saponification value 163 to 180 Iodine value 120 minimum

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ASSESSMENT: ISOPROPYL LINOLEATE 53 assumed ingredient concentration. Isopropyl Linoleate is used in 21 cosmetic products at concentrations ranging from > 0.1% to 10.0% (Table 21.“’ Isopropyl Linoleate has been approved for use as a cosmetic ingredient in Japan.‘g’ This ingredient is not included in the list of substances that may not be used in cosmetic products marketed in countries of the European Economic Community.“”

TOXICOLOGY

Acute Oral Toxicity

The acute oral toxicity of 10.0% Isopropyl Linoleate in corn oil was evaluated using 30 (six groups of 5) male and female fasted, albino rats (weights 200-300 g). The six groups received doses of 2.0, 4.0, 8.0, 16.0, 32.0, and 64.0 cc/kg, respectively, via esophageal catheterization. The following observations were made over a period of two weeks: unkempt coats (8.0 cc/kg); slight diarrhea and unkempt coats (16.0 cc/kg); and wet, oily coats, nasal hemorrhage, and severe diarrhea (32.0 and 64.0 cc/kg). Toxic effects were not observed in animals that received 2.0 and 4.0 cc/kg doses. There were no deaths in any of the groups tested.‘“’

Ocular Irritation

The ocular irritation potential of Isopropyl Linoleate (undiluted and 10.0% aqueous suspension) was evaluated in two studies using male New Zealand albino rabbits (six per concentration tested). The test substance was instilled (0.1 ml) into the conjunctival sac of one eye of each animal; the eyes were not rinsed. Untreated eyes served as controls. Ocular irritation reactions were scored on days 1, 2, 3, 4, and 7 post- instillation according to the scale by Draize. A’~) 0 to 110. In both studies, Isopr‘opyl Linoleate was classified as a slight ocular irritant.(13’ In another study, theocular irritation potential of 10.0% Isopropyl Linoleate in corn oil was evaluated using six albino rabbits. The test substance (0.1 ml) was instilled into the conjunctival sac; the eyes were not rinsed. Reactions were scored over a period of 7 days (at 1 h, 24 h, etc.) according to the Draize scale: 0 to 110. Ocular irritation was not observed in any of the animals tested.“”

Skin Irritation

The skin irritation potential of Isopropyl Linoleate (undiluted and 10.0% aqueous suspension) wasevaluated using six male New Zealand albino rabbits (2.5-3,5 kg). The

TABLE 2. PRODUCTFORMULATION DATA”’

No. of product formulations Total no. of Total no. within each concentration range i%) formulations containing Product category in category ingredient >5-10 >I-5 >o. I-I

Eye and facial makeup products 1358 9 2 7 Face and skin care products, 1293 7 1 3 3 including suntan preparations Miscellaneous hair care products 1027 5 3 2

1988 TOTALS 21 1 8 12

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test substance (0.5 ml) was applied via gauze pads to the left flank (intact) and right flank (abraded) of each animal. Each pad was covered with an occlusive patch and secured with hypoallergenic adhesive plaster. Sites were scored 24 and 72 h after patch application according to the scale: 0 (no edema, erythema) to 4 (severe edema, erythema). In the first experiment, 10.0% isopropyl Linoleate was a nonirritant (Pri- mary Irritation Index = 0.21), and undiluted Isopropyl Linoleate, a slight irritant (Primary irritation Index = 1.13). In the second experiment, 10.0% Isopropyl Linoleate (Primary Irritation Index = 0.96) and undiluted Isopropyl Linoleate (Primary Irritation Index = 1 .OO) were classified as slight irritants. The samples tested in the second experiment were purer than those tested in the first experiment.(13’ In another study, the skin irritation potential of 10.0% Isopropyl Linoleate in corn oil was evaluated using six albino rabbits (3 males, 3 females). The test substance (0.5 ml) was applied to abraded and intact dorsal skin that had been clipped free of hair. Each test site was covered with a patch (two layers of light gauze) that was secured with adhesive tape, and the entire trunk was wrapped with a plastic trunk band. The animals were immobilized during the 24 h exposure period. Reactions were scored at the time of patch removal and 72 h later according to the following scales: 1 (very slight erythema) to 4 (severe erythema to slight eschar formation); 1 (very slight edema) to 4 (severe edema, raised more than 1 mm and extending beyond area of exposure). The final score represented an average of the scores recorded at 24 and 72 h. Skin irritation reactions were not observed in any of the animals tested.‘“’ Isopropyl Linoleate, undiluted and as 10.0% aqueous suspension, was applied daily to the right and left flanks (shaved skin) of each of three male albino rabbits (2.5-3.5 kg) 5 days per week for a period of 8 weeks. Excess test substance was wiped from the skin with gauze 30 set after each application. Application sites were scored daily according to the scale: 0 (no erythema) to 4 (serious erythema with slight scar formation) and 0 (no edema) to 4 (serious edema, covering all of the treated area, or more). Scores were averaged on a weekly basis, and the maximum irritation index (IIMM) was determined at the end of the treatment period. Microscopic examinations were performed on skin samples (2 per animal) that differed macroscopically. The criteria for classifying a substance as poorly tolerated after repeated applications were as follows: (1) macroscopic examination must show pathologic reactions in all three animals treated, (2) pathologic reactions must appear over the entire treated surface of the epidermis, and not only at localized points, and (3) microscopic examination oftwo fragments of treated skin from each rabbit must confirm the macroscopic observations. In the first experiment, 10.0% Isopropyl Linoleate was relatively well tolerated (IIMM = 1.66) and undiluted Isopropyl Linoleate was very poorly tolerated (IIMM = 3.00; treatment was discontinued after 5 weeks). In the second experiment, 10.0% Isopropyl Linoleate was relatively well tolerated (IIMM = 1 .OO) and there was a slight intolerance to undiluted Isopropyl Linoleate (IIMM = 2.66). The samples tested in the second experiment were purer than those tested in the first experiment.‘5’

SUMMARY

Isopropyl Linoleate is the ester of isopropyl alcohol and linoleic acid. In cosmetics, it is used as a skin conditioning agent-emollient at concentrations ranging from 0.1 to 10.0%. In an acute oral toxicity study, none of the albino rabbits that received doses of 10.0% Isopropyl Linoleate in corn oil, 2.0 to 64.0 cc/kg, died.

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ASSESSMENT: ISOPROPYL LINOLEATE 55

Isopropyl Linoleate (undiluted and 10.0% suspension) was classified as a slight ocular irritant when instilled into the eyes of albino rabbits. Following the instillation of 10.0% Isopropyl Linoleate in corn oil, no ocular irritation reactions were observed in albino rabbits. In a primary skin irritation study involving New Zealand albino rabbits, undiluted Isopropyl Linoleate and 10.0% Isopropyl Linoleate were classified as a nonirritant and slight irritant, respectively. When the experiment was repeated using purer samples, undiluted Isopropyl Linoleate and 10.0% Isopropyl Linoleate were both classified as slight irritants. In another primary skin irritation study, 10.0% Isopropyl Linoleate in corn oil did not induce reactions in any of the albino rabbits tested. In albino rabbits, repeated applications of undiluted Isopropyl Linoleate and 10.0% Isopropyl Linoleate were very poorly tolerated and relatively well tolerated, respectively. When the experiment was repeated using purer samples, 10.0% Isopropyl Linoleate was relatively well tolerated and there was a slight intolerance to undiluted Isopropyl Linoleate.

DISCUSSION

Section 1, paragraph (p) of the CIR Procedures states that “A lack of information about an ingredient shall not be sufficient to justify a determination of safety.” In accordance with Section 30(j)(2)(A) of the CIR Procedures, the Panel informed the public of its decision that the data on Isopropyl Linoleate are insufficient to determine whether this ingredient, under each relevant condition of use, is either safe or unsafe. The Panel released a Notice of Insufficient Data Announcement on April 23, 1990 outlining the data needed to assess the safety of Isopropyl Linoleate. The types of data required include: (1) Human skin irritation and sensitization data and (2) genotoxicity data. No offer to supply the skin irritation, skin sensitization, or genotoxicity safety test data was received. However, a UV absorption curve indicating no absorbance in the UVB band was received. In accordance with Section 45 of the CIR Procedures, the Expert Panel will issue a Final Safety Evaluation Report-Insufficient Data. When the requested new data are available, the Panel will reconsider the Final Report in accordance with Section 46 of the CIR Procedures, Amendment of a Final Report.

CONCLUSION

The CIR Expert Panel concludes that the available data are insufficient to support the safety of Isopropyl Linoleate as used in cosmetic products.

ACKNOWLEDGEMENT

This report was prepared by Wilbur Johnson, Jr., Senior Scientific Analyst and Writer.

REFERENCES

1. ESTRIN, N.F., HAYNES, C.R., and WHELAN, J.M. (1982). CosmeticIngredfent Descriptions. CTFA CompendiumofCosmetic Jngredrent Composition. Washington, DC: Cosmetic, Toiletry and Fragrance Association, Inc.

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2. ESTRIN, N.F., HAYNES, C.R., and WHELAN, J.M. (1982). Cosmetic lngredlent Specifications. CTFA Compendium oi Cosmetic ingredient Cornpositron. Washington, DC: The Cosmetic, Toiletry and Fragrance Association, Inc. 3. ESTRIN, N.F., CROSLEY, P.A., and HAYNES, C.R. (1982). CosmeticIngredient Dictionary. Washington, DC:TheCosmetic, Toiletry, and Fragrance Association, Inc., p. 138. 4. VAN DYK CO., INC. (1990). Submission of unpublished data by CTFA. UV spectral analysis on isopropyl linoleate.’ 5. CUILLOT, J.P., CIAUFFRET, J.Y., MARTINI, MC., CONNET, J.F., and SOULE, C. (1981). Raw materials used in cosmetology. Informations Chimie 213, 239-49. 6. NIKITAKIS, I.M. (Editor]. (1988). CFTA Cosmeticlngredrent Handbook. First Edition. Washington, DC: TheCosmetic, Toiletry and Fragrance Association, Inc., p. 237. 7. FOOD AND DRUG ADMINISTRATION (FDA). (1988). Cosmetic product formulation data on isopropyl linoleate. FDA computer printout. 8. CODE OF FEDERAL REGULATIONS (CFR). (Revised as of April 1, 1984). Title 21 Fart 720.4. Voluntary filing of cosmetic product ingredient and cosmetic raw material composition statement. Information requested about cosmetic products. Washington, DC: Government Printing Office, 9. NIKKOL CHEMICALS CO., LTD. (1989). Cross Reference Index: An Jndex Cross-referencing the English Names of Cosmetic Ingredients in /apan with CJFA Adopted Names. Tokyo, Japan: Nikko1 Chemicals Co., Ltd. 10. DUPUIS, 1. (1989). The European Economic Community (EECI Cosmetics Directive. Updated Version-incorporating all amendments up to March 15, 1989. 11. BIO-TOXICOLOGY LABORATORIES, INC. (1975). Submissionof unpublished data byCTFA. Eye irritation, primary irritation, and acute oral LD,, toxicity studies on isopropyl linoleate.’ 12. DRAIZE, J.H. I1 959). Scaleforscoringocular lesions. In: AppraJsalofthe SafetyofChemicals m Foods, Drugs, andCosmetics. Austin, TX: The Association of Fond and Drug Officials of the United States, p. 51. 13. GUILLOT, I.P., GIAUFFRET, J.Y., MARTINI, M.C., GONNET, I.F., and SOULE, C. (1980). Safetyevaluationofcosmetic raw materials: results obtained with 160 samples from various ortgin. pp. 105-108, 109, 110, and 123.

‘Available for review: Director, Cosmetic Ingredient Review, 1101 17th St., N.W., Suite 310, Washington, DC 20036.

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JOURNAL OF THE AMERICAN COLLEGE OF TOXICOLOGY Volume 1, Number 4, 1982 Mary Ann Liebert, Inc., Publishers

Final Report on the Safety Assessment of Cetearyl Octanoate

Cetearyl Octanoate is the esterification product of 2-ethylhexanoic acid and cetearyl alcohol. The acute oral LD50 for Cetearyl Octanoate is estimated from studies with rats to be greater than 8.0 ml/kg. The ingredient produced no significant acute, subchronic or dermal skin or eye irritation when tested in rabbits. The ingredient produced no evidence of skin sensitization in the guinea pig. Similar studies with product formulations containing Cetearyl Octanoate confirmed these results, as well as indicated the ingredient was not phototoxic. In clinical studies, four of 100 subjects showed slight to moderate irritation with undiluted Cetearyl Octanoate. Product formulations containing between 0.2% and 30% Cetearyl Octanoate were tested on a total of 644 subjects with no signs of skin sensitization, photocontact allergenicity, or phototoxicity. From the available information, it is concluded that Cetearyl Octanoate is safe as a cosmetic ingredient in the present practices of use.

CHEMISTRY

Structure and Physical Properties

ETEARYLOctanoate is the ester of cetearyl alcohol, a mixture of fatty alcohols which consists C predominantly of cetyl and stearyl alcohols, and 2-ethylhexanoic acid. The ingredient is a mixture of branched chain fatty esters which conform to the general formula:

0 II CH3(CH2)3CH-C-O-(CH&CH3 I CH2CH3

where n is predominantly 15 and 17.“-3’ Other names include CetyVStearyl 2-Ethylhexanoate, PCL Liquid, and PurCellin Oil. Cetearyl Octanoate is commercially produced by catalytic esterification with removal by azeotropic distillation. (I) It is also prepared by blending cetyl octanoate and stearyl octanoate in a weight ratio of 7:2, which gives some indication of its ester composition.‘4’ The ingredient is a clear, oily liquid with properties similar to those of the oil secreted by the preen glands of waterfowl. Is) It is soluble in absolute alcohol and most nonpolar organic solvents and insoluble in water.‘*’ The available chemical and physical properties of Cetearyl Octanoate are listed in Table 1.

Reactivity

No specific information was available on the reactivity of this ingredient, but it can be expected to undergo chemical or enzymatic hydrolysis to 2-ethyl-hexanoic acid and the corresponding alcohols.

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COSMETIC INGREDIENT REVIEW

TABLE 1. CHEMICALAND PHYSICALPARAMETERSOFCETEARYLOCTANOATE.~

Solid Ester Acid Iodine Saponification point Sp. gr. Refr. ind. value value value value

-4.O”C 0.852 1.444 135 1.0 0.0 135 to to to maximum l.O”C &57 1.446 160 :pa It0 20°C 20°C

aFrom Refs. 1, 2.

Transesterification and other typical ester reactions may also occur. All of the esters present are saturated compounds and would not be expected to autooxidize readily.

Analytical Methods

Positive identification can be made through close matching to standard IR spectra with no indication of foreign materials.“) Hashimoto et a1.‘6’ describe a method for thin-layer chromatography applicable to the subject ingredient .

Impurities

The industry’s purchasing specifications for Cetearyl Octanoate include the following maximum allowable concentrations of chemical impurities:“’ HeavyMetal(asPb)...... 10ppmmaximum Arsenic (as As) ...... , ...... 0.3 ppm maximum

USE Purpose in Cosmetics

Cetearyl Octanoate is a synthetic mixture of fatty acid esters that resembles the preen gland secretion of aquatic birds; thus, it imparts water repelling characteristics to cosmetic formulations. It is also used as an agent that improves “spreadability” and as a “refatting material for dry skin conditions.“(5)

TABLE 2. PRODUCTFORMULATIONDATA.~

Cosmetic product Concentration b No. of product type (Percent) formulations Cetearyl Octanoate Bath oils, tablets and salts > 1.0-5 1 Other bath preparations > 1.0-5 1 Eyeliner >O.l-1 3 Eye shadow >O.l-1 1 so.1 7 Eye makeup remover > 5.0-10 1 Mascara >O.l-1 15 so.1 3 Other eye makeup preparations > 1.o-5 1 >O.l-1 1

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TABLE 2. (Continued). Cosmetic product Concentration b No. of product type (Percent) formulations Hair conditioners > 1.o-5 4 >O.l-1 1 Hair sprays (aerosol fixatives) > 1.o-5 1 Rinses (noncoloring) > 1.o-5 3 >O.l-1 1 Other hair preparations SO.1 1 Blushers (all types) > 5.0-10 > 1.o-5 ; Face powders >O.l-1 12 Foundations > 1.0-5 2 Lipstick > 1.o-5 2 Makeup bases > 1.0-5 11 Other makeup preparations > 1.o-5 4 >O.l-1 1 Douches > 5.0-10 1 Feminine hygiene deodorants >O.l-1 1 Preshave lotions (all types) > 1.o-5 2 SO.1 1 Cleansing (cold creams, cleansing > lo-25 1 lotions, liquids, and pads) > 1.0-5 >O.l-1 : Face, body, and hand (excluding > 1.o-5 8 shaving) preparations >O.l-5 1 Moisturizing > 5.0-10 3 > 1.o-5 11 >O.l-1 2 Night >5.0-10 1 > 1.o-5 4 Paste masks (mud packs) >O.l-1 1 Other skin care preparations > lo-25 1 > 1.o-5 3 Suntan gels, creams, and liquids >l.o-5 2 so.1 1

aFrom Ref. 7. bPreset concentration ranges in accordance with federal filing regulations [21 CFR 720.4(d)(i)].

Table 2 lists product types and the number of product formulations that contained Cetearyl Oc- tanoate as of 1976.“’ Voluntary filing of such information by cosmetic manufacturers conforms to the prescribed format of preset concentration ranges and product types as described in 21 CFR part 720.“) In 1976, Cetearyl Octanoate was reported as an ingredient in 135 cosmetic formulations; two products contained between 10% and 25% Cetearyl Octanoate, while all others used less than 10%.

Surfaces to which Commonly Applied

Products containing this ingredient are applied to all areas of the skin, hair, nails, and mucous membranes. They are most often applied to the face and around the eyes. These formulations may be applied several times a day and may remain in contact with the skin for variable periods of time after each application. Daily or occasional use may extend over many years (Table 2).

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BIOLOGICAL PROPERTIES

Absorption, Metabolism, Storage, and Excretion

Although no specific information on Cetearyl Octanoate was available, comparison to similar long chain fatty acid esters suggests that it would be hydrolyzed in the gastrointestinal tract to 2-ethylhexanoic acid and the corresponding alcohols. These products, in turn, would enter their respective metabolic pathways.

Animal Toxicology

Acute Studies Oral toxicity In each of two studies,‘9.10’ young adult albino rats were fasted for 24 h and given a single administration of undiluted Cetearyl Octanoate by gavage. They were then allowed free access to food and water for 14 days. In the first study, t9) Cetearyl Octanoate was administered to four groups of 10 rats each in doses of 3.98, 5.01, 6.31, or 8.00 ml/kg. There were no deaths and no reported signs of toxicity. In the other study,(lo) 10 rats received 5 ml/kg each. None died, but all showed signs of decreased activity, ataxia, and diarrhea. From these data, the acute oral LD50 for Cetearyl Oc- tanoate is estimated to be greater than 8.0 ml/kg. Acute oral toxicity tests were conducted on product formulations containing 2.5%~“) and 5.0070”~’ Cetearyl Octanoate. The product containing 5.0% was diluted by 50% with volatile silicone, giving an effective concentration of 2.5% Cetearyl Octanoate. In each test, 15 ml/kg doses were administered to a group of 10 adult mice. None of the animals died. Dermal toxicity The acute dermal toxicity of Cetearyl Octanoate was studied on six rabbits.19) The undiluted test material was applied to the intact and abraded clipped skin of the trunk and held in contact for 24 h. The animals were divided into four groups: Group I-2 control rabbits received no test material Group II-2 rabbits received 3.9 ml/kg Group III-2 rabbits received 6.0 ml/kg Group IV-2 rabbits received 9.4 ml/kg After 24 h, the exposed areas were scored on a Draize scale of O-4 for erythema and edema. The animals were further observed for two weeks. Initial gradings ranged from 0 to 1.5, and it was concluded that the material produced only a mild irritation which disappeared in all dose levels by Day 10. There were no deaths and no changes in urine, blood morphology, or gross appearance. An aerosol spray product formulation containing 25-30% Cetearyl Octanoate in its concentrate was tested for acute dermal toxicity on a group of five male and five female albino rabbits.r’3) Five animals received epidermal abrasions, and five retained intact skin. The product was sprayed on the test site for 10 seconds (1.5-2.6 g/kg), and the site was covered with an occlusive bandage for 24 h. There were no detectable skin reactions, toxic and/or pharmacologic effects, or mortality in any of the animals. Primary skin irritation The primary skin irritancy potential of Cetearyl Octanoate was tested in two studies by a Draize single insult patch test technique. In each study, 1.5 ml samples of undiluted Cetearyl Octanoate were applied to the clipped intact and abraded skin of six albino rabbits and occluded for 24 h. The patch sites were evaluated according to the Draize scale following patch removal and again at 72 h. The Primary Irritation Indices in the two tests were 0.019) and 0.17,‘L4) indicating Cetearyl Octanoate to be a nonirritant. Draize primary skin irritation tests were performed on product formulations containing 2.5%~“~) and 5.0%“6) Cetearyl Octanoate. The product containing 5.0% was diluted by 50% with volatile

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ASSESSMENT: CETEARYL OCTANOATE silicone, giving an effective concentration of 2.5% Cetearyl Octanoate. Each study used 0.5 ml doses applied to the abraded and intact skin of three albino rabbits. The formulation containing 2.5% Cetearyl Octanoate produced slight erythema with a Primary Irritation Index of 0.5, whereas the product containing 5.0%, diluted with silicone, produced no signs of irritation. Eye irritation Modifications of the Draize rabbit eye irritation procedure were used in two separate studies. In each, undiluted Cetearyl Octanoate was instilled in one eye of each rabbit; the untreated eye served as control. Grading according to the Draize scale for ocular lesions was conducted at 24,48, and 72 h and again at seven days. In the first study, w three groups of three animals each received 0.1 ml samples of the undiluted test material: Group I-3 animals’ treated eyes remained unwashed Group II - 3 animals’ treated eyes were washed with 20 ml lukewarm water 2 set after instillation Group III - 3 animals’ treated eyes were washed with 20 ml lukewarm water 4 set after instillation There was no eye irritation in Group I, but the treated eyes in Groups II and III showed mild irritation. The total Ocular Irritation Index (011) for all nine animals was 0.78 at 24 h and 0.0 thereafter. In the other study,“‘) the test material was administered to six animals as a two-second spray held six inches from the rabbit’s eye. The 011 was 0.0; no ocular reactions were observed. Since the maximum total score on the Draize scale is 110, these studies indicate Cetearyl Octanoate to be nonirritating to the rabbit eye. The Draize eye irritation test or a modification of it was performed on three product formulations containing 2.5% (I*) 5.0%,‘*9) and 25-30%(20.2’) Cetearyl Octanoate. The product containing 5.0% was diluted by 50% with volatile silicone, giving an effective concentration of 2.5% Cetearyl Oc- tanoate. No significant ocular irritation was produced by any of the formulations. Mucous membrane irritation The potential for mucous membrane irritation by a feminine deodorant aerosol product, the concentrate of which contained 25-30% Cetearyl Octanoate, was tested in two studies with hamsters and dogs. In the first study,“‘) a two-second spray was administered from a distance of six inches directly on the everted left cheek pouch of each of four Golden Syrian hamsters. The untreated right cheek pouch served as a control. The formulation was administered once daily for three consecutive days under Diabutal anesthesia. No irritation was seen in any of the pouches. In the other study,(23) the aerosol formulation was administered as a two-second spray into the vaginal vaults of two dogs for four consecutive days. The vaginal mucosa was biopsied and examined for gross signs of irritation four hours after the fourth treatment. No signs of irritation were observed, and the vaginal tissue appeared grossly normal. Phototoxicity - A guinea pig phototoxicity test was performed on a moisturizer lotion containing 3.2% Cetearyl Octanoate. W) Each of three animals received 0.1 ml of the moisturizer and 0.05 ml of Oxsoralen, a positive control, on different test sites. The animals were exposed to 60 minutes of UV range A (320-400 nm) light 15-20 min after application, and irradiated and nonirradiated test sites were evaluated for erythema. The product formulation produced no signs of phototoxicity. Inhalation Ten adult rats were placed in a 100 1 inhalation chamber and exposed for I h to a concentration of 30 mg/l of an aerosol formulation containing 1.9-2.2% Cetearyl Octanoate.(2s) The rats were removed from the chamber and examined for signs of toxicity immediately and daily thereafter for 14 days. No toxic signs were seen at any time during the test. The incidence of gross lesions seen at autopsy was comparable in treated and control animals. Subchronic Studies Skin sensitization The sensitization potential of Cetearyl Octanoate was tested with the Landsteiner and Jacobs guinea pig sensitization technique. w The backs and flanks of nine white male guinea pigs were clipped free of hair, and a 0.1% solution of Cetearyl Octanoate in Olive Oil USP was injected in-

Supplement Panel Book 1 Page 19 Alkyl Esters Supplement Book 1 COSMETIC INGREDIENT REVIEW tracutaneously three times weekly for a total of 10 injections. The first injection consisted of 0.05 ml, whereas the remaining nine were 0.1 ml each. A challenge injection of 0.05 ml of freshly prepared solution was administered just below the sensitization area two weeks after the tenth sensitization injection. The challenge site was evaluated 24 h later and compared with similar readings taken after the earlier injections. There were no signs of sensitization. A moisturizer lotion containing 3.2% Cetearyl Octanoate was tested according to the Landsteiner and Jacobs guinea pig sensitization technique. lZ6)Groups of eight animals received topical applications of the product formulation, a 0.9 percent saline solution (negative control), or 0.1% dinitrochlorobenzene in acetone (positive control). The moisturizer lotion produced no evidence of skin sensitization. Dermal toxicity Cetearyl Octanoate was applied daily in doses of 1 .O, 2.0, or 4.0 ml/kg by gentle inunction to the clipped intact and abraded skin of albino rabbits. 19)Two animals were used at each dose level and two served as controls. Applications were made five days a week for a total of 20 applications in four weeks. Observations were made daily on body weight, food consumption, and behavior. Urine and blood samples were analyzed at the beginning and end of the study. Cetearyl Octanoate was only very mildly irritating to the skin. The urine and blood data showed no significant deviations from normal, and no other toxic effects were observed. A dermal toxicity study was conducted in which 0.25, 0.5, 1.O, or 2.0 ml/kg of undiluted Cetearyl Octanoate was applied by gentle inunction to the skin of rabbits daily for 90 days. 19)Three animals were used at each dose level; one received epidermal abrasions. Autopsies of all animals were performed at the end of the test, and the skin, lungs, spleen, liver, and kidneys were examined for histopathologic changes. Urine and growth response were normal. Hematological findings were normal except in one animal which, upon autopsy, showed a severe heart condition unrelated to the experimental manipulations. There was no relation between the dose level and observed histopathological abnormalities of internal organs. The skin showed changes in the epidermis indicative of mild irritation, but there were no changes in the dermis.

Clinical Assessment of Safety

Skin Irritation A single insult patch test was performed on 100 individuals, 69 women and 31 men, ranging in age from 14 to 72.19) All but two of the subjects were hospital patients suffering from some kind of skin disease, and 92 of the tested individuals had a history of allergy. Approximately 0.5 ml of 100% Cetearyl Octanoate was applied under a patch and left in place for 24 h. Two subjects showed a positive reaction at 24 h; one reaction persisted and one disappeared by 48 h. Two other subjects developed a reaction by 48 h, one moderate and one very mild. Thus, four of the 100 test subjects showed a slight to moderate skin reaction to Cetearyl Octanoate in a 24-hour patch test. A modified Lanman 21-day cumulative skin irritancy patch test procedure was used to evaluate the irritancy potential of a hair preparation containing 0.40% Cetearyl Octanoate.“‘) The test material was applied under occlusive patches on the back for 21 consecutive days, as were both positive and negative control substances. The 13 panelists produced a cumulative score which elicited the classification of “essentially nonirritating.” Skin Sensitization The Draize-Shelanski repeated insult patch test procedure or a modification of it was performed on six cosmetic product formulations containing 0.2-30% Cetearyl Octanoate. The test material was applied three times a week for a total of ten induction applications. After a two-week rest period, a challenge patch was applied and left on for 24, 48, or 72 h. The results and other details of the studies are summarized in Table 3; no reactions indicative of skin irritation or sensitization were observed in a total of 644 subjects. A maximization test was also used to test the sensitization potential of a product formulation containing Cetearyl Octanoate.‘28) One forearm of each of 25 adult subjects was pretreated with 1%

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TABLE 3. HUMAN REPEAT INSULTPATCH TESTS FOR SKIN IRRITATIONAND SENSITIZATION.

Cont. of Induction Cetearyl exposure No. of subjects Octanoate No. of period/Patch reacting during Challenge exposure No. of reactions Product (percent) subjects a (h) induction phase period (h) at Chalienge Ref. Halston Eye Cream 0.2 56 24 0 48 0 30 z Halston Cleaner 1 50 24 0 48 0 Suntan Lotion 2.5 19 24 0 24 0 2 Moisturizing Lotion 3.2 204 48 0 72 0 32 Bath Gel 5 111 48 4 with 48 0 33 doubtful reactions Feminine Deodorant Aerosol 25-30 204 48 0 48 0 34

Qbjects completing study.

Supplement Panel Book 1 Page 21 Alkyl Esters Supplement Book 1 COSMETIC INGREDIENT REVIEW sodium lauryl sulfate. The test material, which contained 0.40% Cetearyl Octanoate, was then applied under an occlusive dressing for five 48-hour exposures. Following a IO-day rest, the opposite upper arm was pretreated with 10% sodium lauryl sulfate, and a challenge patch of the test material was applied for another 48-hour exposure under occlusion. No instances of contact-sensitization from this material were detected in the maximization test. Photocontact Allergenicity A photocontact allergenicity test was conducted on a suntan lotion containing 2.5% Cetearyl Oc- tanoate. 09) Approximately 0.2 ml of the product was applied under an occlusive patch on each forearm of 27 adults. After 24 h, one forearm of each subject was exposed for 15 min to nonerythrogenic ultraviolet radiation at a distance of IO-12 cm from the light source for a total light dosage of 4,400 pW/cm*. The light source produced UV-A radiation with a peak at 360 nm. The other arm served as a nonirradiated control. After skin reactions were scored, the test sites were covered with untreated patches to prevent inadvertent exposure to sunlight. This procedure was repeated three times a week until a total of 10 induction treatments had been made. After a 10-14 day rest period, the subjects were challenged with the test material under patches which were applied to the original contact site and a fresh, adjacent site. The patches were removed after 24 h, and the test sites were irradiated and scored as before. Additional readings were taken at 48 and 72 h after application. The suntan lotion produced slight irritation in four subjects. One of these subjects exhibited a slight delayed reaction after the sixth exposure of the induction phase at both control and irradiated sites; however, no further reactions were observed at either site. Two subjects showed slight reactions, one after the ninth and one after the 10th induction exposure, but neither subject reacted to the challenge. The fourth subject reacted with erythema after the eighth exposure, but the reaction dissipated after the site was irradiated. This subject also showed a slight erythema after challenge. The investigator concluded that none of the reactions was significant enough to show evidence of photoallergy. Phototoxicity A suntan lotion containing 2.5% Cetearyl Octanoate was tested for phototoxicity on 10 subjects who ranged in age from 22 to 49 years. ‘29) The inner aspects of the forearms were scrubbed with alcohol and tape-stripped 6-10 times to remove several layers of cornified epithelium. About 0.2 g of the product was applied under an occlusive patch to each test site and left in place for 24 h. One forearm of each subject was then exposed to UV light for 15 min at a distance of lo-12 cm from the light source, for a total UV light dosage of 4,400 @W/cm *. No reactions occurred, and it was concluded that the product was not phototoxic. The phototoxicity study on a product containing 2.5% Cetearyl Octanoate is not sufficient to assess the ingredient’s potential for phototoxicity when it is present at the greater concentrations (up to 25%) found in some cosmetic products. Nevertheless, clinical experience indicates that it is unlikely Cetearyl Octanoate would produce any such phototoxic reactions.

SUMMARY

Cetearyl Octanoate is the esterification product of 2-ethylhexanoic acid and cetearyl alcohol. It is used in a wide variety of cosmetic products which may be applied to all areas of the body. The acute oral LD50 for Cetearyl Octanoate is estimated from studies with rats to be greater than 8.0 ml/kg. The ingredient produced no significant skin or eye irritation in Draize rabbit irritation tests, and it was not toxic in acute and subchronic dermal toxicity tests with rabbits. The ingredient produced no evidence of skin sensitization in a guinea pig sensitization test. Product formulations containing between 1.9% and 30% Cetearyl Octanoate were also tested in a variety of animal studies. They produced no signs of acute oral toxicity (Cetearyl Octanoate at 2.5%) acute dermal toxicity (25-30%), skin irritation (2.5%), eye irritation (2.5-30%), mucous membrane irritation (25-30%), skin sensitization (3.2%), inhalation toxicity (1.9-2.2%), or phototoxicity (3.2%). In clinical studies, four of 100 subjects showed slight to moderate irritation after a 24-hour skin

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ASSESSMENT: CETEARYL OCTANOATE patch with undiluted Cetearyl Octanoate, while a product containing 0.4% Cetearyl Octanoate was classified as “essentially nonirritating” in a 21-day cumulative skin irritation test. Product formulations containing between 0.2% and 30% Cetearyl Octanoate were tested on a total of 644 subjects with no signs of skin sensitization. Photocontact allergenicity and phototoxicity studies showed no evidence of photoreactivity for a product containing 2.5% Cetearyl Octanoate.

CONCLUSION

From the available information, the Panel concludes that Cetearyl Octanoate is safe as a cosmetic ingredient in the present practices of use.

ACKNOWLEDGMENT

Mr. Jeffrey Moore, Scientific Analyst and writer, prepared the literature review and technical analysis used by the Expert Panel in developing this report.

REFERENCES

1. COSMETIC, TOILETRY and FRAGRANCE ASSOCIATION (CTFA). (April 26, 1979). CTFA Cosmetic Ingredient Chemical Description: Cetearyl Octanoate. 2. ESTRIN, N.F. (ed.). (1974). CTFA Cosmetic Ingredient Descriptions: Purcellin oil, synthetic. Washington, DC: Cosmetic, Toiletry, and Fragrance Association. 3. ESTRIN, N.F. (ed). (1977). C7’FA Cosmetic Ingredienf Dictionary, 2nd ed. Washington, DC: Cosmetic, Toiletry and Fragrance Association. 4. CTFA. (June 10, 1980). Response to CIR notice of insufficient data report on Cetearyl Octanoate. 5. CTFA. (June 26, 1979). Submission of data. Cetearyl and Stearyl Octanoate: Summary of unpublished safety data. * 6. HASHIMOTO, A., HIROTANI, A., and MUKAI, K. (1967). Thin-layer chromatography of a true wax. Nippon Nogei Kagaku Kaishi. 41(4), 139-44. 7. FOOD and DRUG ADMINISTRATION (FDA). (Aug. 31, 1976). Cosmetic product formulation data. FDA Computer Printout. 8. CODE OF FEDERAL REGULATIONS. (1979). 21 CFR 720. 9. KOLMAR RESEARCH CENTER. (1963). Submission by CTFA. The toxicological examination of Pur- Cellin Oil.* 10. FOOD and DRUG RESEARCH LABS (FDRL). (April 7, 1977). Submission by CTFA. Pur-Cellin Oil: acute oral toxicity in rats.* 11. LEBERCO LABS. (Sept. 12, 1978). Submission by CTFA. Acute oral toxicity in mice.* 12. LEBERCO LABS. (Oct. 23, 1979). Submission by CTFA. Acute oral toxicity in mice.* 13. CTFA. (March 6, 1978). Submission of data. Confidential: acute dermal toxicity in the rabbit.* 14. FDRL. (March 31, 1977). Submission by CTFA. Pur-Cellin Oil: primary skin irritation study with rabbits.* 15. LEBERCO LABS. (Sept. 8, 1978). Submission by CTFA. Primary skin irritation in rabbits.* 16. LEBERCO LABS. (Oct. 12, 1979). Submission by CTFA. Primary skin irritation in rabbits.* 17. FDRL. (March 31, 1977). Submission by CTFA. Neo Pur-Cellin Oil: eye irritation test in rabbits.* 18. LEBERCO LABS. (Sept. 12, 1978). Submission by CTFA. Eye irritation in rabbits.* 19. LEBERCO LABS. (Oct. 15, 1979). Submission by CTFA. Eye irritation in rabbits.* 20. CTFA. (May 18, 1973). Submission of data. Confidential: rabbit eye irritation study.* 21. CTFA. (March 6, 1978). Submission of data. Confidential: rabbit eye irritation study.* 22. CTFA. (May 15, 1973). Submission of data. Mucous membrane irritation in the hamster cheek pouch.* 23. CTFA. (May 17, 1973). Submission of data. Vaginal irritation study in dogs.* 24. CTFA. (June 10, 1980). Submission of data. Guinea pig phototoxicity.’ 25. CTFA. (Feb. 27, 1978). Submission of data. Acute inhalation toxicity in the rat.*

*Available upon request: Administrator, Cosmetic Ingredient Review, Suite 810, 1110 Vermont Ave., N.W., Washington, DC 20005.

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COSMETIC INGREDIENT REVIEW

26. CTFA. (June 10, 1980). Submission of data. Guinea pig sensitization.* 21. CTFA. (April 24, 1980). Submission of data. 21-day cumulative patch test for irritancy potential-human.* 28. CTFA. (April 24, 1980). Submission of data. Maximization test for sensitization potential-human.* 29. FDRL. (Nov. 14, 1978). Submission by CTFA. Final Report: clinical safety evaluation of two products.* 30. UCLA. (Jan. 12, 1979). Submission by CTFA. UCLA File No. 1492: Modified Draize-Shelanski-Jordan patch test.* 31. UCLA. (Jan. 15, 1978). Submission by CTFA. UCLA File No. 1138: Modified Draize-Shelanski-Jordan patch test.* 32. CTFA. (June 10, 1980). Submission of data. Summary: human contact sensitization.* 33. TESTKIT LABS. (Dec. 6, 1979). Submission by CTFA. Repeated insult patch test.* 34. CTFA. (Oct. 9, 1972). Submission of data. Confidential: repeated insult patch test.*

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Final Report on the Safety Assessment of Octyl Palmitate, Cetyl Palmitate and Isopropyl Palmitate

The Palmitates used in cosmetic products are esters of palmitic acid and octyl, cetyl, or isopropyl alcohol. The acute oral LD50 is estimated from studies with rats to be greater than 14.4 g/kg for Cetyl Palmitate and greater than 64.0 g/kg for Octyl and Isopropyl Palmitates. Acute studies with rabbits showed no evidence of dermal toxicity for any of the Palmitates. Isopropyl Palmitate was “well tolerated” and Octyl Palmitate was nontoxic in separate subchronic dermal studies. Rabbit skin tests with the Palmitates showed that they are nonirritating and nonsensitizing. Also, Draize rabbit eye irritation tests on the Palmitates produced either no or only very slight ocular irritation. One of three formulations containing Octyl Palmitate at concentrations between 40% and 50% produced mild irritation. Formulations containing Cetyl Palmitate at concentration of 2.7% were minimally irritating and produced no signs of sensitization, phototoxicity, or photo contact allergenicity. A formulation containing 45.6% Isopropyl Palmitate produced no signs of irritation, sensitization, phototoxicity, or photo contact allergenicity. From the available information, it is concluded that Octyl Palmitate, Cetyl Palmitate, and Isopropyl Palmitate are safe as cosmetic ingredients in the present practices of use and concentration.

CHEMICAL AND PHYSICAL PROPERTIES

he Palmitates are esters of palmitic acid which conform to the general T formula: P CH3(CH2)14C-O-R where R represents the alkyl moiety of Octyl, Cetyl, or Isopropyl alcohol. Because of the technical quality of the palmitic acid and of the alcohols used as industrial starting materials (see Impurities section), the commercially available Palmitates are mixtures of esters conforming to the formula:

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14 COSMETIC INGREDIENT REVIEW

0 R'-;-O-R where R represents the alkyl moiety of one of the aforementioned alcohols, and R’ represents the acyl moiety of palmitic acid. Commercial production methods for most of the Palmitates are considered proprietary information. However, a description of the method of manufacturing a palmitate which is not included in this report”) may also apply to those which are so included. According to this description, Palmitates may be prepared by esterifying the appropriate alcohol with palmitic acid in the presence of approximately 0.1% sulfuric acid. The reaction is performed under reduced pressure, so that the formed water is removed. When this process is completed, the catalyst is neutralized with sodium hydroxide and removed by several water washes. Another ester-yielding reaction which is described in the literature and used in industry is transesterification.‘2’

Octyl Palmitate: Octyl Palmitate is the ester of 2-ethylhexyl alcohol and palmitic acid. It conforms to the formula:(3)

0 II CH$CH ) C-0-CH2-:H(CH2) $ZH3 2 14 CH2CH 3

This ingredient is a clear, colorless, practically odorless liquid. It is soluble in acetone, castor oil, corn oil, chloroform, ethanol, and mineral oil and insoluble in water, glycerin, and propylene glycol. (4) The available chemical and physical parameters of Octyl Palmitate are listed in Table 1.

TABLE 1. Chemical and Physical Parameters of the Palmitates.a

Iodine Acid value value Palmitate Mol. wt. M. P. Sp. Cr. Refr. Ind. Ester value (max.) (max.)

Octyl 388b -c 0.850 1.445 146 to 156 3.0 1 .o to to 0.865 1.4465 @25”C @25T

Cetyl 481b 46T 0.832b 1 .4398b 106 to 120 4.0 1.0 to @25T n,70 54T

Isopropyl 318b 11 T 0.850 1.4355 182.0 1.0 1 .o to to to 0.855 1.4375 191.0 @25OC @25T

aData from Refs. 4, 5, and 21-27. bPure compound. ‘No data.

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ASSESSMENT: OCTYL PALMITATE, CETYL PALMITATE, AND ISOPROPYL PALMITATE 15

Cetyl Palmitate: Cetyl Palmitate is the ester of cetyl alcohol and palmitic acid. It conforms to the formula:(3)

0II CH3 (CH2) 14 -C-O-CH2(CH2)14CH3

Cetyl Palmitate is the chief constituent of commercial purified spermaceti.‘5) It occurs naturally at a concentration of 0.25-0.5 percent in staghorn coral(6) and is biosynthesized by a particular paraffin-oxidizing strain of mycobacterium.“) It is a white, crystalline, wax-like substance which is soluble in alcohol and ether and insoluble in water. (5) Many reports are available concerning the physical chemistry of Cetyl Palmitate in crystalline and liquid states.(*-201 The available chemical and physical parameters of this ingredient are listed in Table 1.

Isopropyl Palmitate: Isopropyl Palmitate is the ester of isopropyl alcohol and palmitic acid. It conforms to the formula:(3)

0 CH3 I CH3 6X2) 14 k-O-0 I CH3

As used in cosmetics, the product is a colorless, almost odorless, mobile Ii- quid mixture of isopropyl esters consisting of a minimum of 60% Isopropyl . Palmitate and lesser amounts of isopropyl laurate, myristate, pentadecanoate, heptadecanoate, and stearate. The ingredient is soluble in acetone, castor oil, chloroform, cottonseed oil, ethyl acetate, ethanol, and mineral oil; it is insoluble in water, glycerin, and propylene glycol. (21a22)One study has explored the crystalline structure of purified Isopropyl Palmitate.(20) The available chemical and physical parameters of Isopropyl Palmitate may be found in Table 1.

Reactivity No specific information on the reactivity of these co,mpounds was found. However, they can be expected to undergo chemical or enzymatic hydrolysis to palmitic acid and the corresponding alcohol. Transesterification and other typical ester reactions may also occur, All of these esters are saturated compounds and would not be expected to autoxidize readily. Analytical Methods

Octyl Palmitate: No specific methods for analysis of Octyl Palmitate were found. As is the case with other Palmitates, however, thin-layer and gas-liquid chromatographies and infrared spectrophotometry techniques would be applicable.

Cetyl Palmitate: Cetyl Palmitate was isolated as a component of spermaceti by high-temperature gas chromatography on a 10% silicone elastomer E301 column. The qualitative composition of spermaceti was determined by thin-layer chromatography. The alkyl ester content was determined by preparative-layer chromatography, and an isolated fraction was identified by infrared spectroscopy.r2*)

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16 COSMETIC INGREDIENT REVIEW

Cetyl Palmitate was also qualitatively determined by thin-layer chromatography using silica plates which contained polyvinyl alcohol. Petroleum ether and 8:2:1 cyclohexane-Et,O-AcOH were used successively as solvents. An intense violet color was developed by spraying the chromatogram successively with a saturated solution of iodine in petroleum ether and water.(zg) Another chromatographic technique involves the use of a silicic acid-impregnated glass-fiber filter paper. Such a method was used to separate Cetyl Palmitate from marine-animal oils.‘30)

Isopropyl Palmitate: Isopropyl Palmitate has been determined by gas-liquid chromatography. Triton X-450 on Fluoroport T and Carbowax 20M on Fluoroport T columns were used for separating the various ingredients of a lipstick that contained Isopropyl Palmitate. Interfering hydrocarbons were removed by column chromatography prior to the GLC analysis.(31)

Impurities The Palmitates used as cosmetic ingredients are mixtures of fatty esters, since the palmitic acid which serves as a starting material is itself a mixture of acids and the alcohols used are mixtures of alcohols. The CTFA Cosmetic Ingredient Chemical Description for palmitic acid lists the following as component acids:(32) Palmitic Acid 80% min. Stearic Acid 11% max. Myristic Acid 7% max. Heptadecanoic Acid 4.5% max. Pentadecanoic Acid 1% max. The CTFA Specification for palmitic acid also includes:(33) Laurie Acid 1.3% max. Oleic Acid 0.4% max. Palmitoleic Acid 0.4% max. Eicosanoic Acid Trace (< O.OSO/,) Myristoleic Acid Trace (< O.OSO/,) Palmitic acid may contain unsaponifiable material, mostly hydrocarbons, at a maximum concentration of 0.3%, and some grades may contain glyceryl monopalmitate at a maximum concentration of 0.07%. Butylated hydroxytoluene (BHT) may be present as an added antioxidant.‘32) Thus, the Palmitates used as cosmetic ingredients may be mixtures of compounds. For example, the CTFA Specification for Isopropyl Palmitate indicates that it consists of a minimum of 60% Isopropyl Palmitate and lesser amounts of isopropyl laurate, myristate, pentadecanoate, heptadecanoate, and stearate.t21)

Octyl Palmitate: Free fatty acids, mainly palmitic acid, are present at a maximum concentration of 1.4%. No known diluents, solvents, or additives are present.‘26)

Cetyl Palmitate: Free fatty acids, mainly palmitic acid, are present at a maximum concentration of 1.8%. No known diluents, solvents, or additives are present. (23) One manufacturer stated that the Cetyl Palmitate used in a particular skin

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ASSESSMENT: OCTYL PALMITATE, CETYL PALMITATE, AND ISOPROPYL PALMITATE 17

moisturizer formulation contains at least 90% Cetyl Palmitate, with the remaining material consisting predominantly of cetyl stearate.(23) The CTFA Specification for cetyl alco.hol includes the following impurities:(34) Hydrocarbons 1.5% max. Ash 0.05% max. ‘Lead (as Pb) 20 ppm max. Arsenic (as As) 3 ppm max.

Isopropyl Palmitate: No known nonester impurities, diluents, solvents, or additives are present.‘25’ The ester composition is varied according to the requirements of an ingredient’s specific intended usage. However, such specifications must conform to the following limits.‘21) Isopropyl Palmitate:

Not less than 65.0% (limits: f 5.0%)

Isopropyl Stearate:

Not more than 35.0% (limits: f 5.0%) Isopropyl Laurate, Myristate, Pentadecanoate, and Heptadecanoate:

None more than 35.0% (limits: * 2.0% each)

USE

Purpose in Cosmetics The Palmitates have a wide spectrum of uses in cosmetic products. All are liquids except the cetyl derivative; they serve as emollients with such specific secondary functions as those of a solvent, plasticizing agent, and/or glossing agent.c2) The Palmitates are efficient opacifiers in cream and lotion shampoos.(35’

Octyl Palmitate: The only specific mention in the literature of the purpose of Octyl Palmitate in cosmetics is that it serves as an emollient.(36)

Cetyl Palmitate: Cetyl Palmitate not only functions as an emollient, but it also contributes a specific body and texture to the majority of cream and lotion products. (2) It functions as a base for ointments, cerates, and emulsions. It is also used in the manufacture of candles, soaps, etc.f5)

Isopropyl Palmitate: Isopropyl Palmitate functions as a physical stabilizer in antiperspirant sticks. (37) It also functions as an emollient, emulsifier, film former, spreader, and solvent in creams, lotions, and eye makeup.(5*36~3B,3g) It is suggested for use in alcohol hair lotions and aftershave preparations.(22) Three publications specifically discuss the use of Isopropyl Palmitate in cosmetics.‘40-42)

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18 COSMETIC INGREDIENT REVIEW

Scope and Extent of Use in Cosmetics

Table 2 lists product types and the number of product formulations reported for each preset concentration range. (43) The cosmetic product formulation computer printout which is made available by the Food and Drug Administration (FDA) is compiled through voluntary filing of such data in accordance with Title 21 part 720.4 of the Code of Federal Regulations (1979). Ingredients are listed in prescribed concentration ranges under specific product type categories. Since certain cosmetic ingredients are supplied by the manufacturer at less than 100% concentration, the value reported by the cosmetic formulator may not necessarily reflect the true, effective concentration found in the finished product; the effective concentration in such a case would be a fraction of that reported to the FDA. The fact that data are only submitted within the framework of preset concentration ranges also provides the opportunity for overestimation of the actual concentration of an ingredient in a particular product. An entry at the lowest end of a concentration range is considered the same as one entered at the highest end of that range, thus introducing the possibility of a two- to ten-fold error in the assumed ingredient concentration. The concentrations that are used range to greater than 50% for Octyl and Isopropyl Palmitate, but to only 10% for Cetyl Palmitate. While these ingredients are most commonly used in the > 1 to 5% concentration range, Isopropyl Palmitate is also frequently employed in the >25 to 50% range. The Palmitates are employed in formulations applied to all those areas of the human integument that are in contact with or in close proximity to all body orifices. They are widely used in lipstick formulations and in products applied close to the eye. Formulations containing the Palmitates may be applied several times a day and may remain in contact with the skin for variable periods of time following each application. Daily or occasional use may extend over many years.

TABLE 2. Product Formulation Data.a

Ingredient/ Concentration No. of product Cosmetic product type (W formulations

Octyl Palmitate Bath oils, tablets, and salts >5-10 1 >l-5 6 Eyeshadow >25-50 2 >5-10 3 >l-5 19 Eye makeup remover ’ >50 1 Other eye makeup preparations >50 1 > 25-50 1 Perfumes >50 1 >l-5 2 Powders (dusting and talcum) >O.l-1 3 (excluding aftershave talc) Other fragrance preparations >l-5 1 Blushers (all types) >50 2 > 25-50 1 > lo-25 2 >5-10 6 >l-5 3

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TABLE 2. (Continued.)

Ingredient/ Concentration No. of product Cosmetic product type m formulations

Face powders >5-10 5 >l-5 9 Foundations >l-5 8 SO.1 Lipstick >5-10 11 11-5 40 Makeup bases 21-5 8 Other makeup preparations >l-5 1 Other personal cleanliness ,1-s 2 products >O.l-1 1 Aftershave lotions >l-5 1 Preshave lotions (all types) >l-5 2 Cleansing (cold creams, >l-5 cleansing lotions, liquids, and pads) Face, body, and hand >lO-25 1 (excluding shaving >5-10 5 preparations) >l-5 10 SO.1 5 Moisturizing >lO-25 3 >5-10 >l-5 13 >O.l-1 3 SO.1 1 Night >lO-25 2 >5-10 1 >l-5 16 >O.l-1 1 Skin fresheners >l-5 4 Other skin care preparations >lO-25 1 >l-5 6 Suntan gels, creams, and >5-10 5 liquids >l-5 3

Cetyl Palmitate Eyeshadow >l-5 7 Sachets >l-5 6 Tonics, dressings, and other >l-5 1 hair grooming aids Blushers (all types) >l-5 23 >O.l-1 1 Foundations >l-5 12 Lipstick >5-10 1 >l-5 289 Makeup bases >l-5 15 Other makeup preparations >l-5 4 Deodorants (underarm) >l-5 1 Other personal cleanliness >l-5 2 products Cleansing (cold creams, >l-5 3 cleansing lotions, liquids, and pads) Face, body, and hand >l-5 1 (excluding shaving preparations)

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20 COSMETIC INGREDIENT REVIEW

TABLE 2. (Continued.)

lngredien t/ Concentration No. of product Cosmetic product type (W formulations

Moisturizing >l-5 >O.l-1 Night >l-5 >O.l-1 Other skin care preparations >l-5

Isopropyl Palmitate Lotions, oils, powders, and >l-5 1 creams >O.l-1 2 Bath oils, tablets, and salts >50 8 >25-50 13 >lO-25 14 >5-10 1 >l-5 1 >O.l-1 5 Bath capsules >25-50 3 Other bath preparations >50 2 >lO-25 1 >5-10 2 >l-5 2 so.1 1 Eyeshadow >5-10 4 >l-5 12 Eye makeup remover >25-50 1 >5-10 2 10.1 1 Other eye makeup preparations >lO-25 5 >5-10 1 Colognes and toilet waters 10.1 2 Perfumes >50 2 >25-50 46 >lO-25 8 >5-10 1 >l-5 1 Powders (dusting and talcum) >5-10 1 (excluding aftershave talc) >O.l-1 9 Sachets >5-10 1 >l-5 5 >O.l-1 1 Other fragrance preparations >25-50 37 >lO-25 8 >5-10 2 >l-5 1 >O.l-1 2 SO.1 1 Hair conditioners >l-5 Hair sprays (aerosol fixative) >l-5 >O.l-1 Permanent waves >O.l-1 Rinses (noncoloring) >lO-25 2 Shampoos (noncoloring) >O.l-1 1 Tonics, dressings, and other >l-5 1 hair grooming aids Wave sets >l-5 2 Other hair preparations >l-5 Hair bleaches >O.l-1

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TABLE 2. (Continued.)

Ingredient/ Concentration No. of product Cosmetic product type fW formulations

Blushers (all types) > 25-50 5 > lo-25 2 >5-10 1 >l-5 9 Face powders >l-5 30 >O.l-1 1 Foundations > 25-50 2 >5-10 9 >l-5 15 >O.l-1 2 Lipstick > lo-25 16 >5-10 67 >l-5 135 Makeup bases > lo-25 1 >5-10 2 >l-5 1 Rouges >25-50 1 Other makeup preparations > 1 O-25 1 >5-10 1 >l-5 3 >O.l-1 1 Cuticle softeners >I-5 1 Nail polish and enamel >l-5 2 removers >O.l-1 1 Deodorants (underarm) >5-10 1 >l-5 7 >O.l-1 1 Feminine hygiene deodorants >l-5 9 >O.l-1 3 Other personal cleanliness >5-10 1 products >l-5 11 >O.l-1 3 Aftershave lotions >l-5 1 >O.l-1 1 Preshave lotions (all types) > lo-25 1 Shaving cream (aerosol, >O.l-1 1 brushless, and lather) Cleansing (cold creams, >50 cleansing lotions, liquids, >25-50 1 and pads) > 1 O-25 2 >5-10 10 >l-5 6 Face, body, and hand >50 1 (excluding shaving > 1 o-25 4 preparations) 25-10 6 >l-5 34 >O.l-1 16 Foot powders and sprays >O.l-1 3 Hormone > lo-25 1 >5-10 2 >l-5 1 Moisturizing > 25-50 1 > 1 O-25 3 >5-10 16 >l-5 23 >O.l-1 11 SO.1 1

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22 COSMETIC INGREDIENT REVIEW

TABLE 2. (Continued.)

Ingredient/ Concentration No. of product Cosmetic product type (96) formulations

Night > lo-25 2 >5-10 9 >l-5 5 >O.l-1 1 Skin fresheners >5-10 1 Wrinkle smoothing (removers) >l- 5 2 Other skin care preparations >50 2 Suntan gels, creams, and >25-50 1 lotions > lo-25 4 >5-10 2 >O.l-1 1

aData from Ref. 43.

BIOLOGICAL PROPERTIES

With respect to pharmacokinetics, pharmacodynamics, teratology, mutagen- esis, and carcinogenesis, there was no reported information concerning any of the Palmitates.

Secondary Effects

When petroleum jelly was applied to human skin at 20 mg/cm2, there was a nearly 100% decrease in skin respiration as measured by both oxygen uptake from and the CO1 discharge to the surrounding air. In contrast, Octyl Palmitate added to petroleum jelly at 10% concentration allowed normal gas-exchange when similarly applied to the skin.(44) Polar alcohols with low molecular weights penetrated cadaveric human abdominal skin (in a diffusion chamber) more rapidly from such nonpolar vehicles as Isopropyl Palmitate than they did from saline solutions.‘45)

Absorption, Metabolism, Storage, and Excretion

Palmitic acid is a component of most animal fats, comprising up to 50% of them. The acid is normally metabolized by P-oxidation or stored in fat depots, as are most of the alcohol moieties after they are oxidized to fatty acids. The terminal groups of the iso-alcohols may yield acetone for excretion or for further metabolism.(46) When fed at a dietary level of 20% to mature male rats, Cetyl Palmitate was quantitatively excreted in the feces.(47)

Animal Toxicology Acute Studies

Oral toxicity Octyl PaImitate: The acute oral toxicity of 100% Octyl’ Palmitate was evaluated in three separate studies. (48-50) In each study, young adult albino rats of both sexes were fasted for 24 hours and given a single administration of Octyl Palmitate by gavage. They were then allowed free access to food and water for

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ASSESSMENT: OCTYL PALMITATE, CETYL PALMITATE, AND ISOPROPYL PALMITATE 23

two weeks. In one study involving 10 rats, the maximum single dose administered was 8 ml/kg. (481 There were no deaths and no apparent signs of systemic toxicity. In another study, Octyl Palmitate was administered to groups of five rats each in doses of 2.5, 5.0, 10.0, 20.0, or 40.0 ml/kg.(4g) No mortalities resulted, and no information regarding other signs of toxicity was reported. The third study used dosage levels of 2.0, 4.0, 8.0, 16.0, 32.0, and 64.0 ml/kg, with five rats at each dose.(50) The animals dosed at 16.0 ml/kg and below showed no apparent toxic symptoms. At 32.0 ml/kg, wet, unkempt coats and mild diarrhea were noted on the first day only. At 64.0 ml/kg, slight ocular hemorrhage and moderate diarrhea were observed. The coats on these animals appeared wet and unkempt for five days following the administration of Octyl Palmitate. No deaths were recorded during the 14-day observation period. No post-mortem or histopathology examinations were reported for any of the three studies.(4g,50) Acute oral toxicity studies similar to those described above were conducted on Octyl Palmitate and four formulations containing Octyl Palmitate, but no details of the procedure or results were reported.(51-55) These studies allow one to estimate that for rats the acute oral LD50 of Octyl Palmitate is greater than 64.0 ml/kg. Cetyl PaImitate: The acute oral toxicity of Cetyl Palmitate was evaluated in four separate studies. (56-5g) In each, adult albino rats were given a single administration of diluted or undiluted Cetyl Palmitate via gavage and then allowed free access to food and water. All animals were fasted overnight prior to administration and observed for two weeks post-dosage. Gross necropsies were performed following the 1Cday observation period, but no results were submitted. In one study, a 5% w/w dispersion in mineral oil was administered to 10 rats (4M 6F) (=) The dosage level was 5 g of the dispersion per kg. One male died betkeen’ Days 7 and 14. Another study(57’ used undiluted Cetyl Palmitate which had been melted prior to being used. Ten rats (5M, 5F) received single doses of 5 ml/kg; one death occurred within 24 hours. In the third study, groups of five male rats each were given 0.464, 1 .OO, 2.15, 4.64, or 10.00 g of Cetyl Palmitate per kg, delivered as a 50% w/v suspension in corn oil. (W There was no mortality. In the fourth study, doses of 5.00, 7.12, 10.14, and 14.43 g of Cetyl Palmitate per kg were administered as a 50% slurry in corn oil to 10 rats at each dosage level.(5g) Diarrhea in one rat at each level was observed three hours following administration. No deaths were reported. Two moisturizer formulations containing Octyl Palmitate were tested for acute oral toxicity. (l) In each test, each of 10 rats (5M, 5F) received a single oral dose of the preparation via gavage. The product containing 2.5% Cetyl Palmitate was given at 30 g/kg, while that containing the ingredient at 2.7% was given at 25 g/kg. No deaths or toxic signs were reported in either study. From these data, it can be estimated that the acute oral LD50 of Cetyl Palmitate in rats is greater than 14.4 g/kg. isopropyl Palmhate: The acute oral toxicity of undiluted Isopropyl Palmitate was evaluated in five separate studies. (5s.60-63)In each, young adult albino rats of both sexes were given single doses of Isopropyl Palmitate via gavage and then allowed free access to food and water. All animals fasted overnight prior to administration of the test material and were observed for two weeks post-dosage. Some studies included gross necropsy following the ICday observation period. Doses of 5.0 ml/kg were administered to 10 rats in each of two studies.(58~“0’ Other than diarrhea and unkempt fur for 36-48 hours following intubation, no

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24 COSMETIC INGREDIENT REVIEW symptoms of toxicity were evident. There were no deaths, and the rats appeared grossly normal for the remainder of the study. In another study, there was no mortality in a group of 10 rats given 8.0 ml/kg doses of Isopropyl Palmitate.(61) A different study also produced no mortality when 20.0 ml/kg doses were administered to 10 rats.(62) In this instance, two males and three females had diarrhea on Day 1, but on subsequent days all animals appeared normal. Gross necropsy revealed no remarkable results. A fifth study used 30 rats in groups of five at each of six dose levels from 2.0 to 64.0 ml/kg.‘63) Except for slightly im- paired locomotion during the first 24 hours, the animals exhibited no signs of toxicity. All animals were completely normal by the second day. An acute oral toxicity study in rats was conducted on a bath oil formulation containing 45.6 percent Isopropyl Palmitate. (64) Ten rats (5M, 5F) were given single oral doses of 25 ml/kg via gavage. There was some evidence of decreased activity up to four hours after treatment, but all animals appeared normal by 24 hours. From these data, it can be estimated that in rats the acute oral LD50 of Isopropyl Palmitate is greater than 64.0 ml/kg.

Dermal toxicity Octyl Palmitate: An acute dermal toxicity study on Octyl Palmitate was conducted with eight rabbits. (48) The test material was applied to the intact and abraded clipped skin of the trunk and held in place with a plastic sleeve for 24 hours. The animals were divided into four groups: Group I: 2 control rabbits received no test material Group II: 2 rabbits received 3.9 ml/kg Group III: 2 rabbits received 6.0 ml/kg Group IV: 2 rabbits received 9.4 ml/kg After the 24hour exposure, the sleeves were removed and the skin reactions were scored for erythema and edema on a Draize scale of O-4. The animals were further observed for two weeks. Initial.gradings ranged up to a score of two, and it was concluded that the material produced only a mild irritation which disappeared in all dose levels by Day 10. There were no deaths, and Octyl Palmitate produced no toxic effects that might be evidenced by changes in urine, blood morphology, or gross appearance. There was no apparent dose response. Cetyl Palmitate: In an acute dermal toxicity study on Cetyl Palmitate, a 50% slurry of the ingredient in distilled water was used. (5g) After each of 10 rabbits was clipped free of abdominal hair and after five were given epidermal abrasions, a single dose of 2.0 g/kg of the slurry was applied to the exposed areas. These areas were then covered with gauze, and for 24 hours, the trunks were wrapped with an impervious material. At the end of the exposure period, dermal reactions were evaluated according to the Draize technique. The slurry caused only a slight dermal irritation with scores of one and two at 24 hours. No deaths occurred during a 14-day period, and it was concluded that the material is not dermally toxic under the conditions of the test. isopropyl Palmitate: An acute dermal toxicity study on Isopropyl Palmitate was conducted with six albino rabbits (3M, 3F) .(60) The trunk skin was clipped free of hair, and in three animals the epidermis was abraded. The trunks were then enclosed in clear polyethylene sleeves, under which 2.0 ml/kg of Isopropyl Palmitate was deposited onto the skin. Following dosing, the rabbits were immobilized for 24 hours, after which time the sleeves and excess material were

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ASSESSMENT: OCTYL PALMITATE, CETYL PALMITATE, AND ISOPROPYL PALMITATE 25 removed. The skin was cleaned and examined for gross changes. No erythema, edema, or other outward signs of toxicity were noted for two weeks.

Primary skin irritation Octyl PaImitate: The primary skin irritancy potential of Octyl Palmitate was tested in three studies by a Draize single insult patch test technique on the clipped intact and abraded skin of albino rabbits. One-half ml samples of Octyl Palmitate were applied and occluded for 24 hours, after which time the patch sites were evaluated for erythema and edema by the Draize technique. Grading was again performed at 72 hours. In one study with three rabbits, all scores were 0.0.(50’ In the second study, six rabbits produced a Primary Irritation Index (PII) of 0.8.‘65) In the third study, the resultant PII for six rabbits was 1 .6.(48) It was concluded from the results of this last test that Octyl Palmitate is a mild irritant. Cetyl Palmitate: Four separate studies used the Draize primary skin irritation technique to evaluate the irritancy potential of Cetyl Palmitate. In each, samples of diluted or undiluted Cetyl Palmitate were applied under occlusive patches on clipped areas of the intact and abraded skin of six albino rabbits and occluded for 24 hours. Dermal reactions were evaluated at 24 and 72 hours, and Plls were calculated by averaging the means of the 24- and 72-hour readings. In one study, 0.5 ml of a 5% w/w dispersion of Cetyl Palmitate in mineral oil produced a PII of 0.38.(56) The mean score was 0.75 at 24 hours; all scores were 0.0 at 72 hours. In another study, 1 .O ml of a 50% dispersion in distilled water was used, giving an effective dosage of 0.5 g of Cetyl Palmitate. (5g) All reaction scores were 0.0. In the third study, 0.5 g of Cetyl Palmitate moistened with saline produced a PII of 0.17.(58) In the fourth study, Cetyl Palmitate was melted and used as a liquid at 100% concentration.(57) A dose of 0.5 ml produced a PII of 0.4. The mean score at 24 hours was 0.8; all scores were 0.0 at 72 hours. The Draize method for primary skin irritation was used to test two moisturizerformulationscontaining2.5% and 2.7% Cetyl Palmitate, respectively.“’ On each of six rabbits, mild irritation was produced by a single 24-hour exposure under occlusion of 0.5 g of the test material at each patch site. The Plls were 1 .O for the 2.5 percent formulation and 0.9 for the 2.7% formulation. isopropyl Palmitate: The Draize primary skin irritation technique was used to evaluate Isopropyl Palmitate in five separate studies.(58*60*65-67) In each, single doses of 0.5 g or 0.5 ml of the undiluted ingredient were applied under occlusive patches on clipped areas of intact and abraded skin of six albino rabbits. Cutaneous reactions were graded at 24 and 72 hours, and the Plls were calculated by averaging the means of the 24- and 72-hour readings. The Plls in the first four studies were determined to be 0.0,(60) 0.38,(66) 0.6,t5*) and 0.92.‘67) In the fifth study,(65) each of four samples of Isopropyl Palmitate was assayed twice, and the eight resultant Plls ranged from 0.25 (nonirritating) to 1.25 (slightly irritating).

Eye irritation Octyl Palmitate: The Draize rabbit eye irritation procedure was used to evaluate Octyl Palmitate in three separate studies.(48.s0~65) In each, 0.1 ml of undiluted Octyl Palmitate was instilled into one eye of each rabbit, and there was no subsequent washing. The other eye remained untreated and served as a control. In one study, the six animals were evaluated for ocular reactions at 24, 48, and 72 hours after administration; the scoring scale differed from that prescribed

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26 COSMETIC INGREDIENT REVIEW by Draize.(4*) Slight conjunctival redness was observed in two animals at 24 hours, but no overall value was computed. If the data obtained at 24 hours are used to calculate according to the Draize technique, the resultant Ocular Irrita- tion Index (011) is 0.33 out of a possible total of 110. In the second study, three rabbits were used and the computed 011 was 2.0; f50) the only reported reaction was conjunctival redness. In the third study, six animals were used and an 011 of 4.17 was determined.(65) These values indicate that Octyl Palmitate is minimally irritating and does not cause any significant injury to the rabbit eye mucous membrane. Cetyl Palmitate: In four separate studies, the Draize eye irritation technique was used to evaluate Cetyl Palmitate. (+s-~~) In each, 0.1 ml or 0.1 g of the test material was instilled into the conjunctival sac of one eye in each of six rabbits. All eyes remained unwashed, and the contralateral eyes served as controls. The treated eyes were examined and graded on the Draize eye irritation scale at 24, 48, and 72 hours. In one study, the Draize 011 for a 5% w/w dispersion of Cetyl Palmitate in mineral oil was 0.0 for all days. (M) The Draize scores for 100% Cetyl Palmitate in another study were 0.3 on the first day and 0.0 thereafter.t5’) In the third study, Cetyl Palmitate delivered as a white powder received scores of 2.3 on the first day, 0.7 on the second day, and 0.3 on the third day.(5g) The 011s for undiluted Cetyl Palmitate in the fourth study were 6.7 on the first day, 2.2 on the second day, and 0.0 on the third day. t5*) It can be concluded from these data, that Cetyl Palmitate is minimally irritating to the rabbit eye. isopropyl Palmitate: The Draize rabbit eye irritation procedure was used to evaluate Isopropyl Palmitate in five separate studies.(58~60,66~68)In each, 0.1 ml of undiluted Isopropyl Palmitate was instilled into the conjunctival sacs of adult albino rabbits. One eye was treated in each animal, while the other remained untreated and served as a control. Irritative effects were scored at 24, 48, and 72 hours according to the method of Draize. In one study, nine rabbits were divided into three groups of three animals each.‘68) While the treated eyes of group one remained unwashed, those of the other two groups were washed with 20 ml of lukewarm water, one at two seconds and the other at four seconds after the test material had been instilled. There were no signs of irritation, and all scores were 0.0. In another study, the eyes of six rabbits remained unwashed after treatment, and the Draize 011 was 0.0.(60) The six rabbits tested in the third study gave an 011 of 0.3 out of a possible total of 110 at 24 hours.(S*) At the 48- and 72-hour gradings, all scores were 0.0. In the fourth study, six rabbits were used, and 011s of 2.33, 0.67, and 0.33 were obtained at 24, 48, and 72 hours, respectively.(66) The fifth study was conducted on four samples of Isopropyl Palmitate.(6s’ The resultant 011s ranged from 3.33 to 6.50 at 48 hours. (An 011 of less than 10 on the Draize scale indicates that the compound does not cause any significant injury to the rabbit eye mucous membrane.)

Inhalation Isopropyl Palmitate: For one hour, ten adult male and female albino rats were exposed to an aerosol of Isopropyl Palmitate in an inhalation chamber.(60) The spray was directed away from the nasal and ocular areas and circulated throughout the chamber for the duration of the exposure. The maximum aerosolization of 200 mgll achieved prior to the test was maintained throughout

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ASSESSMENT: OCTYL PALMITATE, CETYL PALMITATE, AND ISOPROPYL PALMITATE 27 the exposure period. The test animals did not exhibit any toxic symptoms during the 14-day post-exposure observation period; nor did gross autopsies produce any significant findings. The acute toxic dose by inhalation was shown to be greater than 200 mgll of air.

lntraperitoneal injection Isopropyl Palmitate: The lowest published lethal dose for intraperitoneal injection of Isopropyl Palmitate in the mouse is 100 mg/kg.c6”l

Subchronic Studies

Oral toxicity Cetyl PaImitate: Mature rats were fed diets containing 20% Cetyl Palmitate for nine days. The ingredient was quantitatively excreted in the feces, and no abnormalities were noted for the duration of the study.‘46’

Skin sensitization Octyl Palmitate: The Landsteiner and Jacobs guinea pig sensitization technique was used in two separate studies to determine the sensitization potential of Octyl Palmitate. (48.so)The only significant difference between the studies involved the vehicles that were used; in one, Octyl Palmitate was dissolved in Olive Oil USP(48) while in the other, Octyl Palmitate was suspended in sterile, pryrogen- free, physiological saline. t50) In each, 10 white male guinea pigs had their backs and flanks clipped free of hair. A 0.1% solution of Octyl Palmitate was injected intracutaneously three times weekly until a total of 10 injections had been made. While the first injection consisted of 0.05 ml, the remaining nine were 0.1 ml each. Two weeks after the tenth sensitization injection, a challenge injection of 0.05 ml of freshly prepared solution was made slightly below the sensitization area. The challenge site was evaluated 24 hours later, and readings from it were compared with ones taken after the earlier injections. In both studies, investigators concluded that Octyl Palmitate is not a sensitizer. Cetyl Palmitate: In a guinea pig sensitization study, a 1% Cetyl Palmitate suspension in Mazola corn oil was applied topically to ten albino guinea pigs three times per week until a total of 10 sensitizing treatments were made.(sg) While the first treatment consisted of topical application of 0.05 ml of the solution, the remaining nine were made with 0.1 ml volumes. A challenge dose of 0.05 ml was administered on the opposite side of the body two weeks after the tenth treatment. Skin reactions were evaluated 24 hours later and compared with earlier readings. The test solution of Cetyl Palmitate was minimally irritating to the skin when applied topically, but it did not appear to be a sensitizing agent in guinea pigs.

Dermal toxicity Octyl Palmitate: Octyl Palmitate was applied daily in doses of 1 .O ml/kg by gentle inunction to the shaved skin of 10 male and 10 female albino rats.fsl’ Ap- plications were made five days a week for a total of 27 applications in six weeks. Daily observations were made regarding general appearance, behavior, and toxicologic signs. Complete gross necropsy, histopathology, and blood test data were obtained at the termination of the study. The mean hematocrit and red blood cell values of male rats treated with Octyl Palmitate were significantly

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28 COSMETIC INGREDIENT REVIEW lower than those of controls. However, “in the absence of any changes in normal relative ratios of formed elements in the bone marrow, the hematologic changes are of doubtful toxicologic significance.“(51) All other data failed to show any adverse changes attributable to the test material, and it was concluded that Octyl Palmitate produced no systemic toxic effects. A 60-day repeated insult dermal toxicity study was conducted on undiluted Octyl Palmitate. (65) The ingredient was applied daily to an 80 cm2/kg area on the shaved back and flanks of each of three albino rabbits. A 5 cm* area remained untreated and served as control. The ingredient was “poorly tolerated” in two of the three rabbits, and histological examination showed congestive dermatitis in three of six biopsies (two from each animal). isopropyl Palmitate: A 60-day repeated insult dermal toxicity study was conducted on four samples of undiluted Isopropyl Palmitate.(65) The ingredient was applied daily to an 80 cm2/kg area on the shaved back and flanks of each of three albino rabbits. A 5 cm2 macroscopic evaluation showed that the ingredient was either “well tolerated” or “relatively well tolerated” with thickening of the skin and/or presence of vesicles in some rabbits. In Some cases, histological evaluation revealed orthokeratosis and slight acanthosis.

Clinical Assessment of Safety

The available clinical data consist of various experiments involving cutaneous application of these ingredients or of product formulations containing them. Ex- cept for an undocumented history of safe cosmetic use, no other clinical safety information was available.

Octyl Palmitate

Primary irritation A 48-hour occlusive human patch test was conducted on four cosmetic preparations containing Octyl Palmitate at various concentrations.(s2-ss) The preparations included an eye makeup product with 40-50% Octyl Palmitate, an eyeshadow with l-5% Octyl Palmitate, and two moisturizing skin care preparations with l-5% Octyl Palmitate. Each preparation was tested on 100 human subjects, and all results were reported as “negative.” An l&day repeated insult patch test was conducted on three antiperspirant sticks, each containing either 45.72% or 46.52% Octyl Palmitate.r7o) These products were tested along with others, in a multiple patch array on the backs of five male and 15 female predominantly white subjects. The test substance was applied to a l-inch square of nonwoven cloth which was held to the skin of the back under an occlusive impermeable plastic tape. The patch remained in place for 24 hours and was reapplied each day at the same site. A 5-point rating scale (O-4) was used in grading the degree of irritation at each reapplication; there were no reactions, and all such graded scores were 0.0. A 21-day repeated insult patch test similar to the test just described was conducted on the backs of 24 subjects. (‘Ill The product tested was an antiperspirant stick containing 42.25% Octyl Palmitate; the formulation’s other ingredients were not reported. The average cumulative score for the 21 days was 2.58 out of a maximum possible 84. A total of seven subjects exhibited signs of irritation after day eight and were given irritation ratings of at least one; 17 subjects did not react.

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Cetyl Palmitate

A number of cutaneous studies have been conducted on a moisturizer preparation containing either 2.5 or 2.7 percent Cetyl Palmitate in alternate formulations.“)

Primary irritation The procedure described by Kligman and Wooding(72) was used to conduct a lo-day primary skin irritation study in which the moisturizer containing 2.7 percent Cetyl Palmitate was tested on 10 normal adult subjects. For 10 days, approximately 0.3 ml of the undiluted material was applied once daily to the same site under an occlusive patch. There was no evidence of primary irritation.“)

Sensitization The Kligman maximization procedure(73’ was used on a total of 50 subjects to test a nonperfumed version of the moisturizer containing 2.5% Cetyl Palmitate. At challenge, most subjects had a slight erythema, which the investigator attributed to the application of sodium lauryl sulfate. There were six cases of definite erythema (score = + 1) at the 48-hour reading, but only one reaction was still evident at 72 hours. According to the investigator, these reactions did not result from contact sensitization. On the basis of the maximization grading scale, he concluded instead that the material was a “weak potential sensitizer” that was “unlikely to present a risk of contact sensitization under conditions of normal intended use.““) The moisturizer containing 2.7% Cetyl Palmitate was used undiluted in 25 subjects according to Kligman’s modified maximization procedure.(73,74) All patch sites were negative, and it was concluded that the material should be considered a “weak potential sensitizer of the lowest grade.““)

Phototoxicity A phototoxicity study with the moisturizer containing 2.7% Cetyl Palmitate was conducted on 10 normal adult subjects. The material was applied undiluted at 5 PI/cm2 under occlusive patches. After six and 24 hours of contact, sites were irradiated with a 150-watt Xenon Solar Simulator that had been fitted with a Schott WG345 filter to eliminate burning rays (total UV-A irradiance = 25-30 mW/cm2). No instances of phototoxicity were reported, and it was concluded that this formulation is “unlikely to present a risk of phototoxicity under conditions of normal intended use.““)

Photo-contact allergenicity A photo-contact allergenicity study with the moisturizer containing 2.7% Cetyl Palmitate was conducted on 25 normal adult subjects. Applications of 5 PI/cm2 under an occlusive patch for 24 hours were followed by irradiation with a Xenon Solar Simulator (25-30 mW/cm’); this procedure was repeated twice a week for a total of six exposures. A challenge was performed 10 days after the last induction exposure. No instances of photo-contact allergenicity were reported, and it was concluded that “the formulation appears to have a low potential for photo-contact allergenicity under conditions of normal intended use.““)

Safety in-use studies A safety-in-use study of the moisturizer containing 2.5% Cetyl Palmitate was conducted on 30 normal women of unreported age and race. Applications of the

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30 COSMETIC INGREDIENT REVIEW undiluted product to the face and periorbital areas were made thrice daily for 28 days. Examinations for facial, periorbital, conjunctival, and mucous membrane irritations were performed at 14 and 28 days. Occlusive patches were applied pre- and post-treatment. There were no instances of irritation, and it was concluded that “the formulation appears to have a low potential for irritation or sensitization under conditions of normal intended use.““) A similar but longer study was conducted on the moisturizer containing 2.7% Cetyl Palmitate and on a nonperfumed version of this formula; 100 subjects and 50 subjects were used in the respective tests. Examinations for irritation were performed at 28 and 56 days. No instances of mucous membrane, periorbital, or conjunctival inflammation were reported. Although in the tests on the perfumed formulation, one subject did demonstrate a minimal facial erythema on Day 28, there were otherwise no instances of facial erythema. Reduction of the frequency of use to twice daily in this individual resulted in no signs of erythema at the 56-day examination. Of the 150 subjects, only two who used the nonperfumed formulation reacted to the post-treatment occlusive patches, one with minimal (+ 1) erythema at 28 and 48 hours and one with doubtful (+) erythema at 24 hours. It was concluded that “the formulation appears to have a low potential for irritation or acquired contact sensitization under conditions of normal intended use.““)

Isopropyl Palmitate

Primary irritation Three separate studies have been conducted to test the irritancy potential of Isopropyl Palmitate by a 24-hour occlusive patch test technique.(75-77) In each, an unspecified amount of the undiluted ingredient was used, and the skin reactions were scored on a scale of O-4. One study employed 20 subjects for each of two different batches of Isopropyl Palmitate. (75) There were no signs of irritation, and all scores were 0.0. In another such study, two different batches of Isopropyl Palmitate produced one irritation score of 0.5 in a total of 40 subjects.(76) In the third primary irritation study, each of four batches of Isopropyl Palmitate was tested on 20 subjects. (“1 Four of the 80 subjects involved in these three studies received irritation scores of 0.5; all other scores were 0.0. In the three studies overall, undiluted Isopropyl Palmitate was applied to 160 subjects, and it produced five irritation scores of 0.5. To test a bath oil formulation containing 45.6 percent Isopropyl Palmitate,‘64) a lo-day primary irritation study was conducted according to the technique described by Kligman and Wooding. (72) For a total of ten days, ten normal adult subjects received 0.3 ml of the undiluted material once daily at the same site under an occlusive patch. No instances of irritation were reported.

Sensitization One hundred two men and women of unspecified race participated in a modified Draize-Shelanski repeated insult patch test which was used to ascertain the irritation and sensitization potential of Isopropyl PaImitate. Approximately 0.1 ml of undiluted Isopropyl Palmitate was dispersed onto an absorbent nonwoven cotton swatch, 20 x 20 mm, which was then applied to the upper back under an adhesive bandage. The patches were applied three times a week for three weeks and were left in place for 24 hours at each application. Reactions

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ASSESSMENT: OCTYL PALMITATE, CETYL PALMITATE, AND ISOPROPYL PALMITATE 31

were scored on a O-4 irritancy scale 24 hours after patch removal. Seventeen days after the last induction patch was removed, a 24hour challenge application of the test material was made; 24 and 48 hours following the removal of these applications, the challenge reactions were graded. Barely perceptible erythema was noted in three subjects after the second induction insult. All other scores in the induction and challenge periods were 0.0, and it was concluded that Isopropyl Palmitate did not demonstrate a potential for inducing allergic sensitization. Kligman’s modified maximization procedure(73,74) was used on 25 subjects to test an undiluted sample of a bath oil formulation containing 45.6% isopropyl Palmitate.r64) All patch tests were negative, and it was concluded that, “based on the maximization grading scale, the material would be considered the lowest grade, weak potential sensitizer and would be unlikely to present a risk of contact sensitization under conditions of normal intended use.”

Phototoxicity A phototoxicity study of a bath oil formulation containing 45.6% Isopropyl Palmitate was conducted on 10 normal adult subjects of unreported age, sex, and color.‘64) Applications of 5 ~l/cm2 of the undiluted test material were made under occlusive patches, and the application sites were irradiated with a Xenon Solar Simulator (25-30 mW/cm2) after six and 24 hours of contact. No instances of phototoxicity were reported, and it was concluded that this formulation is “unlikely to present a risk of phototoxicity under conditions of normal intended use.”

Photo-contact aiiergenicity Twenty-five adult subjects of unspecified age, sex, and race participated in a photo-contact allergenicity study of the same bath oil containing 45.6 percent Isopropyl Palmitate. (64) Applications of 5 PI/cm2 of the undiluted test material were repeated twice’s week for a total of six applications. The test site remained occluded for 24 hours after each application, and the area was irradiated with a Xenon Solar Simulator (25-30 mW/cm2) following the removal of each patch. A challenge application with irradiation was made 10 days after the last induction exposure. No instances of photocontact allergenicity were reported, and it was concluded that this formulation is “unlikely to present a risk of photo-contact allergenicity under conditions of normal use.”

SUMMARY

The Palmitates are esters of palmitic acid and octyl, cetyl, or isopropyl alcohol. They are used in a wide variety of cosmetic products and may be applied to all areas of the skin. The Palmitates would be expected to be nontoxic in view of their hydrolysis to palmitic acid and to the corresponding alcohols. The acute oral LD50 is estimated from studies with rats to be greater than 14.4 g/kg for Cetyl Palmitate and greater than 64.0 g/kg for Octyl and Isopropyl Palmitate. Acute studies with rabbits showed no evidence of dermal toxicity for any of the Palmitates. In a subchronic dermal toxicity study with rabbits, Isopropyl Palmitate was

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32 COSMETiC INGREDIENT REVIEW

“well tolerated.” Another subchronic dermal toxicity study concluded that Octyl Palmitate is nontoxic. There were no deaths in either of the studies. Other rabbit skin tests with the Palmitates showed that they are nonirritating and nonsensitizing. Draize rabbit eye irritation tests on the Palmitates produced either very slight ocular irritation or none at all. The acute toxic dose in rats by inhalation of Isopropyl Palmitate was shown to be greater than 200 mg/l. The lowest published lethal dose for mouse intraperitoneal injection is 100 mg/kg. A number of human skin tests with the Palmitates and with product formulations containing the Palmitates have been conducted. One of three formulations containing Octyl Palmitate at concentrations between 40% and 50% produced mild irritation. Moisturizer formulations containing Cetyl Palmitate at concentrations of 2.5% and 2.7% were minimally irritating and produced no signs of sensitization, phototoxicity, or photo-contact allergenicity. Undiluted Isopropyl Palmitate was minimally irritating and was reported to be a “weak potential sensitizer” of the “lowest grade.” A bath oil formulation containing 45.6% Isopropyl Palmitate produced no signs of irritation, sensitization, phototoxicity, or photocontact allergenicity. No human sensitization or photo-study data were available on Octyl Palmitate, nor was this ingredient tested at greater than 50% concentration for skin irritation. Clinical data on Cetyl Palmitate are limited to testing at a concentration of 2.7%.

CONCLUSION

On the basis of the available information, the Panel concludes that Octyl Palmitate, Cetyl Palmitate, and Isopropyl Palmitate are safe as cosmetic ingredients in the present practices of use and concentration.

ACKNOWLEDGMENT

Mr. Jeffrey Moore, Scientific Analyst and writer, prepared the literature review and technical analysis used by the Expert Panel in developing this chapter.

REFERENCES

1. COSMETIC, TOILETRY and FRAGRANCE ASSOC. (CTFA). (1977,1978). Submission of data by CTFA. Unpublished data on cetyl palmitate. Safety data on moisturizing formulation containing 2.5-2.7 percent cetyl palmitate.* 2. CTFA. (1978). Submission of data by CTFA. CTFA Cosmetic ingredient chemical description for isobutyl palmitate.* 3. ESTRIN, N.F. fed.). (1977). CTFA Cosmetic Ingredient Dictionary, 2nd ed. Washington, D.C.: Cosmetic, Toiletry and Fragrance Association, Inc.

*Available upon request: Administrator, Cosmetic Ingredient Review, Suite 810, 1110 Vermont Ave., NW, Washington, DC 20005.

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4. ESTRIN, N.F. (ed.). (1974). CTFA Standards: Cosmetic Ingredient Specifications, 2-ethyhexyl PaImitate. Washington, DC.: Cosmetic, Toiletry and Fragrance Association. 5. HAWLEY, G.C. (ed.). (1971). The Condensed Chemical Dictionary, 8th ed. New York: Van Nostrand Reinhold Co. 6. LESTER, 0. and BERGMANN, W. (1941). Marine products. VI. The occurrence of cetyl palmitate in corals. J. Org. Chem. 6, 120-2. 7. KRASIL’NIKOV, N.A., KORONELLI, T.V. and BIS’KO, N.A. (1973). Biosynthesis of cetyJ paImitate by a mycobacterium ceroformans culture. Mikrobiologiya 42(6), 1028-31. 8. ALEBY, S., FISCHMEISTER, I. and IYENGAR, B.T.R. (1971). Infrared spectra and polymorphism of long chain esters. IV. Some esters from tetradecanol, hexadecanol, octadecanol, eicosanol, docosanol, and dodecanoic, tetradecanoic, hexadecanoic, octadecanoic, and eicosanic acid. Lipids 6(6), 421-S. 9. ALEXANDER, A.E. and SCHULMAN, J.H. (1937). Orientation in films of long-chain esters. Proc. Roy. Sot. A161, 115-27. lo. ALEXANDER, A.E. and RIDEAL, E.K. (1937). Reaction kinetics in films. The hydrolysis of long-chain esters. Proc. Roy. Sot. A163, 70-89. 11. BRUMMACE, K.G. (1947). An electron-diffraction study of the heating of straight-chain organic films and its application to lubrication. Proc. Roy. SOC. 191A, 243-52. 12. DORSET, D.L. (1976). Dendritic forms of crystalline long-chain lipids. Naturwissenschaften 63(9), 437-8. 13. DORSET, D.L. (1976). Electra-diffraction crystal structure analysis of cetyJ paImitate. Bioorg. Khim. 2(6), 781-8. 14. DRYDEN, J.S. and WELSH, H.K. (1969). Dielectric absorption and molecular motion in aliphatic compounds. Discuss. Faraday Sot. 48, 174-80. 15. HUNTER, J. and EDDY, CR. (1967). Dielectric properties of some long-chain esters in the solid state. J, Am. Oil Chem. Sot. 44(6), 341-3. 16. KOHLHASS, R. (1938). X-ray investigation of definite single crystals of the cetyJ ester of palmitic acid. Z. Krist. 98, 418-38. 17. MACDOUGALL, G. and OCKRENT, C. (1942). Surface-energy relations in liquid/solid systems. I. The adhesion of liquids to solids and a new method of determining the surface tension of liquids. Proc. Roy. Sot. A180,151-73. 18. TAMAMUSHI, B. (19341. The orientation of molecules on surfaces of water and aqeuous solutions. Bull. Chem. Sot. Japan 9, 161-5. 19. THIESSEN, P.A. and SCHOON, Th. (1937). Electron diffraction on natural surfaces of organic single crystals. Z. Physik. Chem. 836, 216-31. 20. VORLANDER, D. and SELKE, W. (1927). The uniaxial structure of soft solid crystalline masses and of crystalline liquids. Z. Physik. Chem. 129, 435-74. 21. ESTRIN, N.F. (ed.). (1974). CTFA Standards: Cosmetic Ingredient Specifications, isopropyl Palmitate. Washington, D.C.: Cosmetic, Toiletry and Fragrance Association. 22. GREENBERG, L.A. and LESTER D. (1954). Handbook of Cosmetic Materials. New York: Interscience Publishers. 23. CTFA. Submission of data by CTFA. (1977,1978). Unpublished data on moisturizing formulation containing 2.5-2.7 percent cetyl paImitate.* 24. CTFA. (1978). Submission of data by CTFA. CTFA Cosmetic ingredient chemical description for cetearyl palmitate.’ 25. CTFA. (1978). Submission of data by CTFA. CTFA Cosmetic ingredient chemical description for cetyl palmitate.* 26. CTFA. (1978). Submission of data by CTFA. CTFA Cosmetic ingredient chemical description for isopropyl palmitate.* 27. WINDHOLZ, M. (ed.). (1976). The Merck Index, 9th ed. Rahway, NJ.: Merck and Co. 28. HONOWAY, P.J. (1968). Chromatographic analysis of spermaceti. J. Pharm. Pharmacol. 20(10), 775-9. 29. MARTINI, M.C. and CHAMBON, M. (1969). Color reactions characteristic of certain poly (vinyl alcohols). Their use in the detection of lipids in chromatography. Ann. Pharm. Fr. 27(2), 143-6. 30. REISER, R., SORRELS, M.F., and BENDER, M. (1957). Chromatographic analysis of some constituents of marine-animal oils. Comm. Fisheries Rev. 19(4a), 9-10. 31. GROSS, F.C. (1966). Gas-liquid chromatography of some fatty acid esters and alcohols in lipsticks. J. Assoc. Off. Anal. Chem. 49(6), 1196-200. 32. CTFA. (1979). Submission of data by CTFA. CTFA Cosmetic ingredient chemical description for palmitic acid.* 33. ESTRIN, N.F. (Editor). (1974). CTFA Standards: Cosmetic Ingredient Specifications, palmitic acid 95 percent. Washington, D.C.: Cosmetic, Toiletry and Fragrance Association. 34. ESTRIN, N.F. (Editor). (1974). CTFA Standards: Cosmetic Ingredient Specifications, cetyl alcohol. Washington, D.C.: Cosmetic, Toiletry and Fragrance Association.

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34 COSMETIC INGREDIENT REVIEW

35. POWERS, D.H. (1972). Shampoos, in: Cosmetics: Science and Technology, 2nd ed. New York: Wiley- Interscience. v. 2. pp. 73-l 16. 36. RlNZLER, C.A. (1977). Cosmetics: What the Ads Don’t Jell You. New York: Thomas J. Crowell CO. pp. 142, 159. 37. PLECHNER, S. (1972). Antiperspirants and deodorants, in: Cosmetics: Science and Technology, 2nd ed. New York: Wiley-Interscience. v. 2. pp. 373-416. 38. BARNElT, C. (1972). Emollient Creams and Lotions, in: Cosmetics: Science and Technology, 2nd ed. New York: Wiley-Interscience. v. 1. pp. 27-104. 39. WETTERHAHN, J. (1972). Eye Makeup, in: Cosmetics: Science and Technology, 2nd ed. New York: Wiley- Interscience. v. 1. pp. 393-422. 40. BERGWEIN, K. (1954). Use of fatty acid esters of low molecular weight alcohols in cosmetics. Fette Seifen Anstrichm. 56, 422-4. 41, HOLZNER, G. (1963). The use of isopropyl fatty esters in cosmetic formulations. Am. Perfum. Cosmet. 78(10), 89-93. 42. TUAN, T.A. (1970). Use of isopropylic esters of some saturated fatty acids in cosmetology. Parfum. Cosmet. Savons 13(6), 454-7. 43. FDA. (Aug. 31, 1976). Cosmetic product formulation data. Washington, D.C.: Food and Drug Administra- tion. 44. JACOBI, 0. (1970). Skin respiration and cosmetics. Am. Perfum. Cosmet. 85(7), 25-30. 45. BLANK, I.H. (1964). Penetration of low-molecular-weight alcohols into skin. I. Effect of concentration of alcohol and type of vehicle. J. Invest. Dermatol. 43, 415-20. 46. HILDITCH, T.P. (1940). The Chemical Constitution of Natural Fats. New York: John Wiley & Sons. 47. CALLOWAY, D.H., KURTZ, C.W., and POTTS, R.B. (1959). Some physiological characteristics of esters of cetyl alcohol. Can. J. Biochem. Physiol. 37(l), 17-23. 48. KOLMAR RESEARCH CENTER. (Aug. 1967). Submission of data by CTFA. Unpublished data on octyl palmitate. The toxicological examination of alpha-ethyl-hexyl-palmitate (Wickenol 155).* 49. FOOD AND DRUG RESEARCH LABORATORIES. (April 6, 1976). Submission of data by CTFA. Unpublished data on octyl paimitate. Acute oral toxicity in rats.’ 50. BIO-TOXICOLOGY LABORATORIES. (July 13, 1972). Submission of data by CTFA. Unpublished data on octyl palmitate. Toxicity studies, 2-ethylhexyl palmitate.* 51. CTFA. (May 21, 1975). Submission of data by CTFA. Unpublished data on octyl palmitate. Safety evaluation of selected raw ingredients.* 52. CTFA. (1975). Submission of data by CTFA. Unpublished data on octyl palmitate. CIR safety data submission: Eye makeup preparation, 40-50 percent octyl paImitate.* 53. CTFA. (1977). Submission of data by CTFA. Unpublished data on octyl palmitate. CIR safety data submission: Eyeshadow (waterproof type), l-5 percent octyl paImitate.* 54. CTFA. (1977). Submission of data by CTFA. Unpublished data on octyl palmitate. CIR safety data submission: Moisturizing skin care lotion, l-5 percent octyl paImitate.* 55. CTFA. (1978). Submission of data by CTFA. Unpublished data on octyl palmitate. CIR safety data submission: Moisturizing skin care cream, l-5 percent octyl palmitate.’ 56. BIOMETRIC. (1977). Submission of data by CTFA. Unpublished data on cetyl palmitate. Standamul 1616.* 57. CONSUMER PRODUCT TESTING. (1977). Submission of data by CTFA. Unpublished data on cetyl palmitate. Standamul 1616.’ 58. ARMAK. (Oct. 25, 1972). Submission of data by CTFA. Unpublished data. Acute toxicity studies of Kessco esters. Bull. t’jo. 73-l.’ 59. MB RESEARCH LABORATORIES. (Oct. 25, 1974). Submission of data by CTFA. Unpublished data on isopropyl palmitate. Report on primary dermal irritation in rabbits.* 60. BIO-TOXICOLOGY LABORATORIES. (1975). Submission of data by CTFA. Unpublished data on isopropyl palmitate. Toxicity studies on isopropyl palmitate.’ 61. KOLMAR RESEARCH CENTER. (June 20, 1972). Submission of data by CTFA. Unpublished data on isopropyl palmitate. The toxicological examination of Wickhen isopropyl palmitate (Wickenol 11 l).* 62. SPRINGBORN INSTITUTE FOR BIORESEARCH. (April 27, 1978). Submission of data by CTFA. Unpublished data on isopropyl palmitate. A study of the acute oral (LD50) toxicity: Starfol IPP.’ 63. BIO-TOXICOLOGY LABORATORIES. (June 23, 1970). Submission of data by CTFA. Unpublished data on isopropyl palmitate. Toxicity study for Emery Industries.’ 64. CTFA. (1978). Submission of data by CTFA. Unpublished data on isopropyl palmitate. Safety data on a bath oil formulation containing 45.6 percent isopropyl palmitate.* 65. GUILLOT, J.P., MARTINI, M.C., and GIAUFFRET, J.Y. (1977). Safety evaluation of cosmetic raw materials. J. Sot. Cosmet. Chem. 28(7), 377-93.

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ASSESSMENT: OCTYL PALMITATE, CETYL PALMITATE, AND ISOPROPYL PALMITATE 35

66. HILL TOP RESEARCH. (July 3, 1968). Submission of data by CTFA. Unpublished data on isopropyl palmitate. Primary skin and acute eye irritation studies on a series of test materials.* 67. MB RESEARCH LABORATORIES. (1977). Submission of data by CTFA. Unpublished data. Waxenol 815.’ 68. LEBERCO LABORATORIES. (Dec. 1, 1975). Submission of data by CTFA. Unpublished data on isopropyl palmitate. Eye irritation study: Emerest 2316.* 69. FAIRCHILD, E.J. (Ed.). (1977) Registry of Toxic Effects of Chemical Substances, vol. II. Cincinnati, Ohio: U.S. Dept. of H.E.W., Public Health Service, Center for Disease Control. 70. CTFA. (Sept. 16, 1977). Submission of data by CTFA. Unpublished data on octyl palmitate. Irritation study: Is-day repeat insult patch test.* 71, CTFA. (May 23, 1977). Submission of data by CTFA. Unpublished data on octyl palmitate. Irritation study: ST-day repeat insult patch test.* 72. KLIGMAN, A.M. and WOODING, W.M. (1967). A method for the measurement and evaluation of irritants on human skin. J. Invest. Dermatol. 49(l), 78-94. 73. KLIGMAN, A.M. (1966). The identification of common contact allergens by human assay. Ill. The maximization test: a procedure for screening and grading contact sensitizers. J. Invest. Dermatol. 47(S), 393-409. 74. KLIGMAN, A.M. and EPSTEIN, W. (1975). Updating the maximization test for identifying contact allergens. Contact Dermatitis 1, 231-9. 75. CTFA. (April 5, 1972). Submission of data by CTFA. Unpublished data on isopropyl palmitate. Clinical evaluation report, profile No. 5087.* 76. CTFA. (Sept. 12, 1973). Submission of data by CTFA. Unpublished data on isopropyl palmitate. Clinical evaluation report, profile No. 42050.* 77. CTFA. (March 9, 1973). Submission of data by CTFA. Unpublished data on isopropyl palmitate. Clinical evaluation report, profile No. 40065.* 78. CTFA. (Sept. 20, 1976). Submission of data by CTFA. Unpublished data on isopropyl palmitate. Allergic contact sensitization test, No. 008-76.’

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Final Report on the Safety Assessment of Decyl and lsodecyl Oleates

Decyl Oleate and lsodecyl Oleate are esters of oleic acid. Decyl Oleate is used in cosmetic products at concentrations ranging from I 0.1 to > 50%. lsodecyl Oleate is used at concentrations of > O.l-25%. Animal studies have shown both Decyl Oleate and lsodecyl Oleate to possess low acute oral toxicities in rats with LD50s of > 40 ml/kg. Single application dermal and eye studies with rabbits have shown these materials at 100% concentrations produce little or no irritation. Daily applications of 15% or 100% concentrations for 60 days to the skin of rabbits produced a moderate degree of irritation with both Decyl and lsodecyl Oleate. Neither of the ingredients was found to be a sensitizer when tested in guinea pigs at concentrations of 15%. Repeated insult patch tests containing l-5% Decyl Oleate showed no signs of sensitization. Testing with formulations containing 5.5% Decyl Oleate produced a low number of reactions in 402 human subjects in the Schwartz- Peck Prophetic Patch Test and 204 subjects with undiluted lsodecyl Oleate on nine subjects showed a total irritation score of 1.0 out of a maximum of 756. It is concluded that, because of both the chemical similarity of these compounds and the similarity of the available animal and human data, Decyl and lsodecyl Oleates warrant a conclusion of safe in the concentrations of present practices and use in cosmetics.

CHEMICAL AND PHYSICAL PROPERTIES

ecyl Oleate and Isodecyl Oleate are esters of oleic acid, Formed by ester- D ification of oleic acid with decyl or isodecyl alcohol, they have the following structural formulas:

Decyl Oleate- CH,(CH2)7CH=CH(CH2),COOCH2(CH2)8CH3

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86 COSMETIC INGREDIENT REVIEW

Methyl Oleate is a compound chemically related to Decyl Oleate and lsodecyl Oleate. Its structural formula is as follows:

CH3(CH ) CH=CH(CH2)7COOCH3 2 7

The safety of methyl oleate is not under review in this report. Information and data pertaining to this compound are included to permit a more complete appraisal of the safety of Decyl Oleate and Isodecyl Oleate. Some of the chemical and physical properties of these esters are given in Table 1 .(lm3)

Reactivity

Unsaturated fatty acids and their esters readily undergo autoxidation.(4’ Methyl oleate can serve as a model for autoxidation reactions which all the oleic acid esters exhibited. This compound undergoes autoxidation to give primarily trans-hydroperoxides,(5) which are highly unstable and readily decompose to keto and hydroxy keto acids. f6) Some hydroperoxides have been found to possess carcinogenic potential. (‘) Methyl oleate undergoes photochemical decomposition in direct sunlight and in the presence of oxygen to form the ozonide of methyl oleic acid. (W The most important secondary products of autoxidation include alpha, beta-unsaturated carbonyl compounds.(5) Hematin compounds,‘9) metals,(lO) and chlorinated hydrocarbon insecticides’“) ac- celerate the autoxidation reaction by shortening the induction period.

Analytical Methods

Methyl oleate can be generated in purities of 98% or better by repeated distillation with urea at a low temperature. (l*) Analysis of this and related compounds is done by gas-liquid or thin-layer chromatography. The position of the double bond can be determined by von Rudolph’s oxidation procedure. Infrared spectroscopy can be used to delineate cis-trans isomers.(12-14) Although gas-liquid chromatography remains the preferred routine analytical method for fatty acid ester mixtures, utilization of high performance liquid chromatography (HPLC) is increasing; for the latter has the advantage of identifying polymerized and oxidized esters which the former does not detect.(15,16)

TABLE 1. Properties.a

Approx. Saponification lodine Acid Value Specific Ingredient M.W. Appearance Solubility value value (Max.) gravity

Decyl Oleate 422 Light yellow Soluble in alcohol 130-142 55-65 5.0 0.855Xl.865 liquid

Insoluble in Hz0

lsodecyl 422 NAb NA 130-l 45 SO-65 5.0 NA Oleate

aData from Refs. l-3. bNA = Not found in available literature.

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ASSESSMENT: DECYL AND ISODECYL OLEATES 87

Impurities

The following impurities have been reported in the oleate esters under review.‘2)

lngredien t Impurity Percent Present Decyl Oleate Oleic Acid 2.5 max. Isodecyl Oleate Oleic Acid 2.5 max.

PURPOSE AND FREQUENCY OF USE IN COSMETICS

Decyl and lsodecyl Oleates have been widely used in cosmetic products. When applied to the skin alone, they deposit a thin oily film that is neither greasy nor tacky. They have good lubrication properties and possess low viscosity.(“) Both materials are used as dispersants and lubricants in cosmetic formulations, and these are particularly important in makeup and makeup removers, in which they are used as wetting agents for iron oxide pigments; particles of such pigment are dispersed and easily suspended. The use of these ingredients facilities the application and removal of a suspension. By virtue of its branched chain structure, lsodecyl Oleate possesses several distinct properties. It has the ability to lower the freezing point of the emulsion phase of products, as well as to control product viscosity. In dispersible bath oils, it forms a white emulsion, giving the tub water a rich and milky appearance. It al!jo has the ability to suspend aluminum chlorohydrate, which makes it valuable for dry antiperspirant formulations. Lipstick formulations have employed Isodecyl Oleate because its coupling properties increase the hardness and strength of the product without reducing its flow characteristics. Table 2 indicates categories of product use and concentrations of use for Decyl and lsodecyl Oleate. (“‘The cosmetic product formulation computer printout which is made available by the Food and Drug Administration (FDA) is com-

TABLE 2. Product Formulation Data.’

Ingredient Concentration No. of product Cosmetic product type I%) formulation5

Decyl Oleate Lotions, oils, powders, and >l-5 1 creams Bath oils, tablets, and salts > lo-25 >5-10 Eyeshadow > lo-25 >l-5 Eye makeup remover >l-5 Mascara >O.l-1 Other eye makeup preparations >50 Hair conditioners >O.l-1 Hair sprays (aerosol >O.l-1 fixatives) Rinses (noncoloring) >O.l-1 Shampoos (noncoloring) >O.l-1 Blushers (all types) >5-10 >l-5

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88 COSMETIC INGREDIENT REVIEW

TABLE 2. (Continued.)

Ingredient Concentration No. of product Cosmetic product type (W formulations

Foundations >5-10 2 >l-5 6 >O.l-1 1 Lipstick >l-5 12 Makeup bases >5-10 1 >l-5 5 Other makeup preparations >l-5 1 Other manicuring preparations >5-10 1 Cleansing (cold creams, > lo-25 2 cleansing lotions, liquids, >5-10 5 and pads) >l-5 1 10.1 2 Face, body, and hand > lo-25 2 (excluding shaving >5-10 3 preparations) >l-5 7 Moisturizing >lO-25 2 >5-10 7 >l-5 12 >O.l-1 2 Night >l-5 1 Other skin care preparations >50 3 >l-5 2 Suntan gels, creams, and >25-50 2 liquids >l-5 2

lsodecyl Oleate Bath oils, tablets, and salts >5-10 1 Other bath preparations >O.l-1 1 Eyeshadow >l-5 8 Blushers (all types) >l-5 1 Foundations >l-5 2 Other makeup preparations >l-5 2 Deodorants (underarm) >l-5 1 Other personal cleanliness >l-5 1 products Cleansing (cold creams, > lo-25 1 cleansing lotion, liquids, and pads) Face, body, and hand >lO-25 1 (excluding shaving >5-10 1 preparations) Moisturizing >5-10 2 >l-5 2

aData from Ref. 18.

piled through voluntary filing of such data in accordance with Title 21 Part 720.4 of the Code of Federal Regulations (1979). Ingredients are listed in prescribed concentration ranges under specific product type categories. Since certain cosmetic ingredients are supplied by the manufacturer at less that 100% concentration, the value reported by the cosmetic formulator may not necessarily reflect the true, effective concentration found in the finished product; the effective concentration in such a case would be a fraction of that reported to the FDA. The fact that data are only submitted within the framework of preset concentration ranges also provides the opportunity for overestimation of the actual concentration of

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ASSESSMENT: DECYL AND ISODECYL OLEATES 89

an ingredient in a particular product. An entry at the lowest end of a concentration range is considered the same as one entered at the highest end of that range, thus introducing the possibility of a two- to ten-fold error in the assumed ingredient concentration. The compounds are employed in a variety of cosmetics, including makeup preparations, skin care preparations, and eye-shadow. Concen- trations of use range from 10.1 to > 50% for Decyl Oleate and > O.l-25% for lsodecyl Oleate. Products containing these two materials are applied with varying frequency to all areas of the skin. In such formulations as blushers and moisturizing creams, exposure may occur several times a day, while in other cases there may be daily (deodorants) or less frequent (rinses, hair conditioners) applications. This occasional or daily use may extend over a period .of years.

BIOLOGICAL PROPERTIES

Absorption, Metabolism, and Excretion

Unsaturated fatty acids such as oleic acid penetrate the skin via the hair follicles and sebaceous glands. (IT) Esters of oleic acid presumably follow a similar route of absorption. A major portion of ingested fatty acids is absorbed across the intestinal mucosa, and the ester of these are completely hydrolyzed by ex- tracellular enzymes in the lumen of the small intestines. After the resulting free fatty acids are taken up by the villa1 cells, they may or may not be esterified to glycerol; they are then packaged into chylomicra. These lipoproteins leave the intestinal mucosal cells, enter the lymphatic system, and ultimately end up in the blood. Capillary endothelial cell-bound lipoprotein lipases degrade the chylomicra, freeing up their fatty acid content for cellular uptake. Depending upon nutritional and hormonal factors, intracellular enzymes then catalyze either the production of triglycerides for lipid storage or the catabolism of the fatty acids via /3-oxidation.(‘9) The absorptive and excretive properties of methyl oleate may be typical of other esters of oleic acid. Thirty to 120 minutes after being fed mixtures of methyl oleate (57-75 mg) that consisted of 10% methyl 9,ll -octadecadienoate (Md-OD), mic:e were sacrificed and the stomach and intestinal contents extracted and mixed with octane. These lipid-bearing extracts were then assayed for Md-OD via ultraviolet absorption spectroscopy at 230.7 nm. The results indicate that the mouse is capable of absorbing 60 mg of the mixture administered in a single dose in a maximum of two hours. The rate of absorption was nearly constant.r20) When rats were fed a diet containing 5% methyl oleate for 12 weeks, there was an increase in the fecal excretion of free fatty acids, hexane-soluble acid materials, and unsaponifiable matter as compared to controls fed fat-free diets.(2’)

Animal Toxicology

Acute Studies

Oral toxicity Decyl Oleate: This ingredient was administered to Wistar rats by intragastric intubation at dose levels of 2.5, 5.0, 10.0, 20.0, and 40.0 ml/kg (three male and

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90 COSMETIC INGREDIENT REVIEW two female rats per dose level). (22) The animals were fasted for 24 hours prior to dosing. All animals were observed daily for 14 days following administration and no deaths were recorded. The acute LD50 of undiluted Decyl Oleate was greater than 40.0 ml/kg of body weight. Wistar-derived rats (groups of five male, five female) were dosed by gavage with either 5.0 g/kg of undiluted Decyl Oleate or 5.0 g/kg of 20 percent Decyl Oleate 80% mineral oil.(23) The rats were fasted for 18 hours prior to dosing. The animals were observed for signs of pharmacologic activity and drug toxicity at 1, 3, 6, and 24 hours post-dosing, after which daily observations were made for a total of 14 days. One death was recorded for male animals in the diluted sample group, and one female died following treatment with the undiluted sample. No treatment-related effects were noted in any of the surviving animals. Examination of tissues of nonsurvivors and survivors at gross autopsy revealed no abnormalities. lsodecyl Oleate: This ingredient was administered to Wistar rats by intragastric intubation at dose levels of 2.5, 5.0, 10.0, 20.0, and 40.0 ml/kg (two female and three male rats per dose level). (24) The animals were fasted for 24 hours prior to dosing. One death was recorded at the highest dose level. The acute LD50 of undiluted lsodecyl Oleate was reported to be greater than 40.0 ml/kg of body weight.

Dermal irritation Decyl Oleate: Draize (25) and Federal Hazardous Substances Labeling Act’12) (FHSA) methods were used to conduct primary skin irritation studies. Test samples of Decyl Oleate (undiluted, 10 percent in corn oil and 20% in mineral oil) were applied (0.5 ml) to clipped areas of intact and abraded albino rabbits skin (six animals in each group). The abrasions were longitudinal, epidermal incisions sufficiently deep to penetrate the stratum corneum, but not so deep as to disturb the dermis. Following application of the test material, the exposed area was covered with a patch and the entire experimental area was sealed with impervious sheeting. The animals were immobilized for a 24-hour period. The mean scores for 24- and 72-hour gradings were averaged to determine final irritation values. The primary irritation index (PII) for undiluted Decyl Oleate was calculated to be 0.28.(2s) It was also determined that Decyl Oleate had primary irritation indices of 0.08 as a 10 percent solution in corn oil(22) and 0.05 as a 20% solution in mineral oil.(25) A modified Draize method was used to conduct primary dermal irritation studies with undiluted and 15% Decyl Oleate diluted in polyoxyethylene sorbitan stearate (3%), a perservative (2%), and water; the material was found to be nonirritating (Table 3).(2”) lsodecyl Oleate: This ingredient (0.5 ml) was applied undiluted to the clipped intact and abraded skin of three albino rabbits. The patch area was covered with Webril patches, and the entire experimental area was sealed with Blenderm Surgical Tape. After the animals had remained in restraining stocks for 24 hours, the patches were removed and the treated skin evaluated according to the method of Draize. The skin was evaluated again 72 hours later and the primary irritation index was calculated. The lsodecyl Oleate was calculated to have a PII of 1 .OO in this experiment. (24) In a summary report it was determined that undiluted lsodecyl Oleate applied to the skin of rabbits had a primary irritation index of 0.28.‘27’

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ASSESSMENT: DECYL AND ISODECYL OLEATES 91

TABLE 3. Ocular and Cutaneous Effects in Rabbits.”

Primary irritation of the irritation Concentration ocular mucous membrane of the skin Compound (W Ollb P/F

Decyl 100 1 h: 2.67 0.13 Oleate 24 h: 3.00 48 h: 0.33 15 0.00 lsodecyl 100 1 h: 4.00 assay 1: Oleate 24 h: 0.00 0.13 48 h: 0.0 assay 2: 0.00 15 assay 1: 0.00 assay 2: 0.00

aData from Ref. 24. bOll = Ocular irritation index; Scoring: O-100. cPll = Primary irritation index; Scoring: < 0.5 = nonirritant, 0.5-2 = slight irritant.

Primary dermal irritation studies were conducted with undiluted and 15 percent lsodecyl Oleate diluted with polyoxyethylene sorbitan stearate (3%), a Ipreservative (2%), and water. A modified Draize method was used, and the material was found to be nonirritating (Table 3).c2@

Eye irritation Decyl Oleate: Decyl Oleate (0.1 ml) was instilled into the right eyes of rabbits at concentrations of 15 percent in corn oil,(2B) 20% in mineral oil and 100°/o.~25~ Under the Draize eye irritation procedure, the animals’ left eyes served as the experimental controls. Primary eye irritation scores were 0.0 for the undiluted material, 0.0 for the 20% in mineral solution, and 1 .O for the 15% in corn oil solution. Employing a modified Draize method, Guillot et al.(26) tested Decyl Oleate at a 100% concentration for ocular irritancy in rabbits. Irritation readings were made at 1 hour and at 1, 2, 3, 4, and 7 days. Scores (presented in Table 3) indicate that the material is at most a very slight irritant. lsodecyl Oleate: Draize eye irritation studies were undertaken with undiluted Isodecyl Oleate and 15 percent lsodecyl Oleate in corn oil. The 15% solution had an eye irritation score of 0.3,(24) while the undiluted solution had a score of 0.0.(27) Using a modified Draize method, Guillot et al.(26) tested lsodecyl Oleate at a 100% concentration for ocular irritancy in rabbits. Irritation readings were made at 1 hour and at 1, 2, 3, 4, and 7 days. Scores (presented in Table 3) indicated that the material is at most a very slight irritant.

Su bchronic

Oral toxicity When fed to five male and five female rats for 12 weeks as 5% of their diet, pure methyl oleate and two other methyl esters of fatty acids caused the females to lose considerably more body weight than the test animals that were fed corn

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92 COSMETIC INGREDIENT REVIEW oil (Table 4). Food intake and food efficiency were not altered, but symptoms of essential fatty acid deficiency (as seen by “scaliness of the tail”) were evident as early as the 7th week of the experiment.(“)

Dermal irritation Decyl Oleate: Decyl Oleate was applied daily for eight weeks to the skin of rabbits; the FHSA method was used with slight modifications. Undiluted or 15% Decyl Oleate diluted in polyoxyethylene sorbitan stearate (3%), a preservative (2%), &d water were used. Reactions were evaluated daily, and at the end of the eight-week period, two skin samples were examined histologically. The compound produced irritation at the 15% and 100% levels (Table 5). Following a seven-day rest period, a challenge patch was applied to determine whether observed reactions were allergic or irritant in origin; the results indicated that they were the latter.(26) The Landsteiner and Jacobs guinea pig sensitization procedure was utilized to evaluate Decyl Oleate at a 15% solution in corn oil.(28) Hair on the backs of 10 white male guinea pigs weighing 300-500 g each was clipped prior to the study’s initiation and whenever it was subsequently necessary. A 0.1% solution of the Decyl Oleate was intracutaneously injected three times a week into the shaved back areas of the animals. These injections, which continued until a total of 10 had been made, consisted of 0.05 ml of test solution for the first injection and 0.1 ml for the remaining nine. Two weeks after the tenth injection, a challenge injection of 0.05 ml of freshly prepared solution was made just below the region of the sensitizing injections. The Control, corn oil (0.1%) in physiological saline, was given in the same amount as the test material. Twenty-four hours following injection, scorings were recorded for the diameter, height, and redness of any reactions, and the average scores for the 10 sensitizing injections were compared with the scores for challenge injections. If the challenge score is substantially higher than the average value, sensitization would be reported. Because Decyl Oleate failed, upon challenge, to exhibit higher values, it is considered to be a nonsensitizer (its score was 0.0).

TABLE 4. Body Weight, Food Consumption and Food Efficiency of Rats Fed Methyl Oleate for 12 Weeks.a

Body weight Food eaten Food efficiency (9, (t?J (gJ

Fat in diet Male Female Ma/e Female Male Female

None 194 146 779 803 0.202 0.140 Methyl Oleate 192 134 688 601 0.234 0.172 (5% of diet) Methyl-l l- 191 139 701 607 0.227 0.178 Eicosenate 6% of diet) Methyl Erucate 170b 138 595 590 0.245 0.186 (5% of diet) Corn Oil 195 157 693 632 0.238 0.192 (5% of diet)

aData from Ref. 21. bAlthough it appears that methyl erucate retarded the growth of the males, statistical analysis showed that this difference was not significant at P = 0.05.

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ASSESSMENT: DECYL AND ISODECYL OLEATES 93

TABLE 5. 60-Day Cumulative Cutaneous Effects in Rabbits.”

60-Day Cumulative irritation Scores

Concentration Macroscopic Histological Interpretation Compound w evaluation evaluation of tolerance

Decyl 100 Poorly tolerated; Pathological reaction: Poor Oleate skin thickening congestive dermis. in the three rabbits, vesicles in one animal. 15 Relatively well No significant patho- Good tolerated; some logical reaction. papulae or vesicles. lsodecyl 100 Poorly tolerated; Congestive dermis with Poor Oleate vesicles in one pericapillary imflam- animal. matory infiltrates; pathological reaction. 15 Relatively well No significant patho- Poor tolerated; logical reaction. episodical macules, papulae and vesicles.

aData from Ref. 26.

lsodecyl Oleate: When lsodecyl Oleate at a 15% concentration was tested according to the Landsteiner and Jacobs guinea pig sensitization procedure, it was determined to be a nonsensitizer (its score was 0.0).(24) lsodecyl Oleate was applied daily for eight weeks to the skin of rabbits; the FHSA method was used with slight modifications. Undiluted or 15% lsodecyl Oleate diluted in polyoxyethylene sorbitan stearate (3%), a preservative (2%), and water were used. Reactions were evaluated daily, and at the end of the eight- week period two skin samples were examined histologically. The compound produced a moderate degree of irritation at the 100% level (Table 5). Following a seven-day rest period, a challenge patch was applied to determine whether observed reactions were allergic or irritant in origin. Results indicate that they were the latter.(26)

Clinical Assessment of Safety

Decyl O/eater A human repeated insult patch test was conducted on 103 subjects with a skin conditioner containing l-5% Decyl Oleate. Patches containing approximately 0.2 ml of undiluted sample were applied on Monday, Wednes- day, and Friday for three consecutive weeks. Fourteen days after the final insult patch, challenge patches containing the undiluted skin conditioner were applied, and results were graded 48 and 96 hours later. No evidence of sensitization was found; no information on irritation potential was reported.(29) Four formulations of a foundation containing Decyl Oleate (5.5%) were tested in the Schwartz-Peck Prophetic Patch Test and the Draize-Shelanski Re- peated Insult Patch Test. “Virtually zero reactions occurred in 402 subjects in the Schwartz-Peck Test and 204 subjects in the Draize-Shelanski Test.“(23) lsodecyl Oleate: A single insult (24-hour) occlusive patch test was conducted on 19 human subjects with undiluted lsodecyl Oleate. The test material did not elicit any erythematous reactions. A summary report of the study concluded that

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94 COSMETIC INGREDIENT REVIEW

Isodecyl Oleate exhibits an acceptably low incidence of primary skin irritation under occlusive patch test conditions.(27) According to an industry raw material evaluation, a procedure was undertaken with Isodecyl Oleate under the conditions of a Maibach-type Cumulative lrritancy Assay. When Isodecyl Oleate was applied undiluted under patch conditions to the skin of nine subjects for 21 consecutive days, it was found to have a total irritation score of 1.0 out of a maximum possible 756.(30)

SUMMARY

Decyl Oleate and lsodecyl Oleate are esters of oleic acid. Decyl Oleate is used in cosmetic products at concentrations ranging from ~0.1 to >50%. lsodecyl Oleate is used at concentrations of > O.l-25%. Animal studies have shown Decyl Oleate and lsodecyl Oleate to possess low acute oral toxicities in rats; both have LD50s of > 40 ml/kg. Single application dermal and eye studies with rabbits have shown that these materials in concentrations up to 100% produce little or no irritation. When 15% or 100% concentrations were applied to the skin of rabbits daily for 60 days, both Decyl Oleate and lsodecyl Oleate produced moderate degrees of irritation. Neither ingredient was found to be a sensitizer when it was tested in guinea pigs at concentrations of 15%. Repeated human insult patch tests on 103 subjects with a skin conditioner containing l-5% Decyl Oleate showed no signs of sensitization. Industrial testing with formulations containing 5.5% Decyl Oleate produced a low number of reactions in 402 human subjects in the Schwartz-Peck Prophetic Patch Test and in 204 subjects in the Draize-Shelanski Patch Test. Repeated insult patch tests with undiluted lsodecyl Oleate on an unspecified number of human subjects showed a total irritation score of 1 .O out of a possible maximum of 756. A single insult occlusive patch test on 19 human subjects with undiluted lsodecyl Oleate produced a low level of primary skin irritation. No chronic, oral subchronic, carcinogenicity, mutagenicity, or teratogenicity animal testing data were available to the Panel. Nor were there any phototoxicity or photosensitization studies in humans.

CONCLUSION

The Panel concludes that because of both the chemical similarity of the compounds and the similarities among the available animal and human data, Decyl and lsodecyl Oleates warrant a conclusion of safe in the concentrations of present practices and use in cosmetics.

REFERENCES

1. ESTRIN, N.F. (Editor). (1974). CTFA Standards: Cosmetic ingredient Descriptions. Washington, DC: Cos- metic, Toiletry and Fragrance Association, Inc. 2. CTFA. (1979). Submission of data by CTFA. Cosmetic ingredient descriptions for Decyl Oleate, lsodecyl Oleate, Oleyl Oleate, Butyl Oleate, Isopropyl Oleate and Methyl Oleate.* 3. WINDHOLZ, M. (ed.). (1976). The Merck Index, 9th ed. Rahway, NJ: Merck.

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ASSESSMENT: DECYL AND ISODECYL OLEATES 95

4. GORTNER, R.A. (1949). Outlines of Biochemistry, 3rd ed. New York: John Wiley and Sons. 5. SWERN, D. and COLEMAN, J.E. (1955). Reactions of fatty materials with oxygen. XX. Recent developments in the autoxidation of methyl oleate and other monounsaturated fatty materials. J. Am. Oil Chem. SOC. 32, 700-3. 6. WHITE, A., HANDLER, P., and SMITH, E. (1973). Principles of Biochemistry, 5th ed. New York: McGraw- Hill Book Co. p. 65. 7. HIATT, H.H., WATSON, J.D., and WINSTEN, J.A. (1977). Origins of Human Cancer Book A: Incidence of Cancer in Humans. Cold Spring Harbor: Cold Spring Laboratory. p. 231. 8. KITAHARA, K. (1961). Unsaturated fatty acids. III. Investigation by gas chromatography of the photochemical conversion of unsaturated fatty acids. Yakugaku Zasshi 81, 126-9. 9. TAPPEL, A.L. (1953). The mechanism of the oxidation of unsaturated fatty acids catalyzed by hematin compounds. Arch. Biochem. Biophys. 44(2), 378-95. 10. YAMADA, T. and MOCIDA, Y. (1969). Effect of metals on the autoxidation of methyl oleate. Kanagawa Daigaku Kogakubu Kenkyu Hokoku 1969(7), 136-45. 11, BENCZE, K. and WILDBRETT, C. (1971). Effect of chlorohydrocarbon insecticides on radical reactions of methyl oleate. Fette Seifen Antrichm 73(3), 157-60. 12. GUNSTONE, F.D., MCLAUCHLAN, J., SCRIMCEOUR, C.M., and WATSON, A.P. (1976). Improved procedures for the isolation of pure oleic, linoleic, and linolenic acids or their methyl esters from natural sources. J. Sci. Food Agric. 27(7), 675-80. 13. ALLEN, R.R. (1969). Determination of trans isomers in GLC (gas-liquid chromatographic) fractions of unsaturated ester. Lipids 4(6), 627-28. 14 LIE KEN JIE, M.S.F. and LAM, C.H. (1976). Fatty acids. IX. the thin-layer chromatographic behavior of all the cis, cis- and tram-, trans-dimethylene-interrupted methyl octadecadienoates and methyl octadecadiynoates. 1. Chromatogr. 124(l), 147-51. 15. SCHOLFIELD, C.R. (1975). High performance liquid chromatography of fatty methyl esters: preparative separations. Anal. Chem. 47(8), 1417-20. 16. SCHOLFIELD, C.R. (1975). High performance liquid chromatography of fatty methyl esters: Analytical separations. J. Am. Oil Chem. Sot. 52, 36-7. 17. STRIANSE, S.J. (1972). Hand creams and lotions, vol. 1, in: Cosmetics: Science and Technology, 2nd ed., 3 vols. MS. Balsam and E. Sagarin (eds.). New York: Wiley-Interscience. pp. 193-95. 18. FDA. (Aug. 31, 1976). Cosmetic product formulation data. Computer printout. Washington, DC: Food and Drug Administration. 19. LEHNINGER, A.L. (1976). Biochemistry, 2nd ed. New York: Worth, 1975. LIE KEN JIE, M.S.F. and LAM, C.H. Fatty acids. IX. The thin-layer chromatographic behavior of all the cis, cis- and trans, trans-dimethylene-interrupted methyl octadecadienoates and methyl octadecadiynoates. J. Chromatogr. 124(l), 147-51, 20. MEAD, J.F., DECKER, A.B., and BENNETT, L.R. (1951). The effect of x-irradiation upon fat absorption in the mouse. J. Nutr. 43, 485-99. 21. MURRAY, T.K., CAMPBELL, J.A., HOPKINS, C.Y., and CHISHOLM. M.J. (1958). The effect of mono-enoic fatty acid esters on the growth and fecal lipides of rats. J. Am. Oil Chem. Sot. 35, 156-58. 22. CTFA. (1976). Submission of data by CTFA. Decyl Oleate, 19.4b 2a.* 23. CTFA. (1975). Submission of data by CTFA. Decyl Oleate, 19.4b 6b.* 24. CTFA. (1977). Submission of data by CTFA. lsodecyl Oleate, 19.4b 2c.* 25. CTFA. (1975). Submission of data by CTFA. Decyl Oleate, 19.4b 3.* 26. GUILLOT, J.P., MARTINI, M.C. and GIAUFFRET, J.Y. (1977). Safety evaluation of cosmetic raw materials. J, Sot. Cosmet. Chem. 28(7), 377-93. 27. CTFA. (1977). Submission of data by CTFA. lsodecyl Oleate, 19.4b 5a.* 28. CTFA. (1976). Submission of data by CTFA. Decyl Oleate, 19.4b 2b.* 29. CTFA. (1976). Submission of data by CTFA. Oleyl Oleate, 19.4b 6a.* 30. CTFA. (1976). Submission of data by CTFA. lsodecyl Oleate, 19.4b 5b.*

- *Available upon request: Administrator, Cosmetic Ingredient Review, Suite 810, 11 10 Vermont Ave., NW, Washington, DC 20005.

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JOURNAL OF THE AMERICAN COLLEGE OF TOXICOLOGY Volume 4, Number 5, 1985 Mary Ann Liebert, Inc., Publishers

Final Report on the Safety Assessment of Butyl Stearate, Cetyl Stearate, Isobutyl Stearate, lsocetyl Stearate, Isopropyl Stearate, Myristyl Stearate, and Octyl Stearate

The 7 Stearates described in this report are either oily liquids or waxy solids that are primarily used in cosmetics as skin emollients at concentrations up to 25 percent. The toxicology of the Stearates has been assessed in a number of animal studies. They have low acute oral toxicity and are essentially nonirritating to the rabbit eye when tested at and above use concentration. At cosmetic use concentrations the Stearates are, at most, minimally irritating to rabbit skin. In clinical studies the Stearates and cosmetic products containing them were at most minimally to mildly irritating to the human skin, essentially nonsensitizing, nonphototoxic and nonphotosensitizing. Comedogenicity is a potential health effect that should be considered when the Stearate ingredients are used in cosmetic formulations. On the basis of the information in this report, it is concluded that Butyl, Cetyl, Isobutyl, Isocetyl, Isopropyl, Myrjstyl, and Octyl Stearate are safe as cosmetic ingredients in the present practices of use.

CHEMISTRY

Definition and Structure

he seven cosmetic ingredients reviewed in this report are listed below in al- T phabetical order: 1. Butyl Stearate 2. Cetyl Stearate

107

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108 COSMETIC INGREDIENT REVIEW

3. lsobutyl Stearate 4. lsocetyl Stearate 5. Isopropyl Stearate 6. Myristyl Stearate 7. Octyl Stearate

The aforementioned Stearates are esters of stearic acid.“) They conform to the general formula:

0 II CH,KH,),aC-OR

where R represents the alkyl moiety of butyl, cetyl, isobutyl, isocetyl, isopropyl, myristyl or octyl alcohol. (‘-‘) The safety of stearic acid as a fragrance raw material and as a food ingredient has been reviewed by the Research Institute for Fra- grance Materials (RIFM)‘4’ and the Federation of American Societies for Experi- mental Biology (FASEB),(5) respectively. A review of the scientific literature from 1920 to 1973 on stearic acid has been published.(6) Because of the technical grade of the stearic acid and the alcohols used as starting materials, the commercially available Stearates are mixtures of esters conforming to the formula:

0 II where R-C represents the acyl moiety present in commercial stearic acid and where R represents the alkyl moiety of 1 of the aforementioned alcohols.(1,2) The chemical names and structural formulas of the 7 Stearates under review are presented in Table 1.

Methods of Manufacture

The Stearates are prepared by the esterification of stearic acid with the appropriate alcohol in the presence of an acid catalyst:

CH,(CHJ,aCOOH + ROH H;t CHKH3,aCOOR + Hz0

Stearic acid Alcohol Stearate ester where R represents the alkyl moiety of butyl, cetyl, isobutyl, isocetyl, isopropyl, myristyl, or octyl alcohol. (1a2*9)The reaction products are refined either by catalyst neutralization, vacuum distillation, or various decolorization-deodorization techniques to remove residual traces of alcohol.(9.10) The higher stearate esters (Isocetyl, Myristyl) can be made by heating with or without acidic catalysts. If an acidic catalyst is used, it is neutralized, and the product is filtered to remove the salts.(9)

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FINAL REPORT:SAFETY ASSESSMENT OF STEARATES 109

Butyl Stearate can be prepared by the reaction of silver stearate with n-butyl iodide at 100°C’“~12) or by the transesterification of glyceryl tristearate (tristearin) with n-butyl alcohol. (12) This ester may also be prepared by the alcoholysis of glycerol stearate @tearin).

Composition and Impurities

Butyl Stearate, as used in cosmetics, is a liquid mixture of butyl esters consisting of a minimum of 90 percent butyl palmitate and butyl stearate, with lesser amounts of butyl laurate, myristate, pentadecanoate, heptadecanoate, and oleate.(13,14) According to cosmetic ingredient specifications published by the Cos- metic, Toiletry and Fragrance Association, (13*14)the ester composition is:

As specified by the buyer; provided that the sum of the specified mean levels equals 100.0 percent and that the individual mean levels and specification limits conform to the following:

Butyl Stearate: Mean Level: Not less than 40.0 $ercent] (Limits: *5.0 [percent])

Butyl Stearate plus Butyl Palmitate: Mean Level: Not less than 90.0 [percent] (Limits: l 5.0 [percent])

Butyl Laurate, Myristate, Pentadecanoate, Heptadecanoate, and Oleate: Mean Level: Not more than 10.0 [percent] each (Limits: h2.0 [percent])

lsocetyl Stearate, as used in cosmetics, is reported to contain a maximum of I percent free fatty acid. (‘) There are no known diluents, solvents, or additives present in Butyl, Isobutyl, Isocetyl, Isopropyl, or Octyl Stearate.(‘s2) The composition and impurities of the various starting materials (stearic acid and butyl, cetyl, isocetyl, isopropyl, and myristyl alcohol) have been reported elsewhere.(13-17)

Properties

Butyl Stearate is a stable, colorless, oily liquid. It is resistant to oxidation and undergoes no appreciable degradation when exposed to a temperature of 205OC for 3 hours. At about 19OC, the ester solidifies. Butyl Stearate is slightly soluble to insoluble in water (0.2 percent) and is soluble in acetone, chloroform, ether, alcohol, ketones, ethyl acetate, aromatic and aliphatic hydrocarbons, fats, waxes, mineral oils, and many plasticizers. (10-14~18-21)Th e chemical and physical properties of this compound have been reviewed by Lower.(21) The infrared spectrum of Butyl Stearate is available in cosmetic specifications published by the Cosmetic, Toiletry and Fragrance Association.(14*22) lsobutyl Stearate is a paraffinlike crystal substance at “low temperature;““” at room temperature, it is a liquid. (9) lsocetyl Stearate occurs as an oily, colorless or yellow liquid with practically no odor. It is soluble in ethanol, isopropanol, mineral oil, castor oil, acetone, and ethyl acetate and is insoluble in water, glycerin,

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TABLE 1. Chemical Names and Structural Formulas’3~‘~8’ ingredient Chemical Names CAS No. Structural Formula

0 II Butyl Stearate Butyl octadecanoate 123-95-5 CHKHJ,aC-O&H9 n-Butyl octadecanoate Butyl octadecylate n-Butyl stearate Octadecanoic acid, butyl ester Stearic acid, butyl ester 0 II Cetyl Stearate Hexadecyl octadecanoate 1190-63-2 CH,(CH,),~C-OCH*(CH~)I~CH, n-Hexadecyl stearate Octadecanoic acid, hexadecyl ester Stearic acid, hexadecyl ester 0 CH, II I lsobutyl Stearate Octadecanoic acid, 2-methylpropyl ester 646-l 3-9 c~a(CHz),sC-OCHLH Stearic acid, 2-methylpropyl ester I Stearic acid, isobutyl ester CH, 2-Methylpropyl octadecanoate

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FINAL REPORT:SAFETY ASSESSMENTOF STEARATE~ 111

b 5 w ,. $0 rd -A a 2 $j -2 p ‘QJm &-$; $X Ffj 7l ,g N 0 r g g 5 Emu$ s g m-c z-o.v’5 L m L1: E;t;$L &OS&

a, 5 w 5 5 z b 2 5 m .-z r ‘z r s

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112 COSMETIC INGREDIENT REVIEW and propylene glycol. (10*17~23)Isopropyl Stearate is a liquid at room temperature.(g) Myristyl Stearate is a waxy solid at room temperature.(g) Chemical and physical data for the Stearate ingredients are presented in Table 2. No data were available for Cetyl Stearate.

TABLE 2. Chemical and Physical Data

Ingredient Reference

Butyl Stearate

Molecular formula LkLOz 8, 11 Molecular weight 340.57 11, 19, 21, 25 Melting point 16OC 11 18.5-20.5”C 21 19.0-2OT 12 19.5”C 18, 25 19.5-2ooc 10 27.0°C 11 27.5’C 8, 19 Boiling point 212-216°C (25 mm) 21 220-225T (25 mm) 18, 19 343T 11 35OT 10 351-360T (760 mm) 21 Viscosity 8 centistokes (25T) 21 Specific gravity 0.851-0.861 (20”/2O”C) 20 0.855-0.860 (25°/200C) 10,12 0.855-0.875 (25”/25”C) 11,19 0.853-0.860 (25°C) 21 Vapor density 11.4 25 Refractive index 1.441 (25°C) 21 1.4430 (20°C) 10,12 1.4328 (50°C) 8 Saponification value 146-177 20 167-173 21 Acid value 1 .O maximum 2, 13, 14, 20 1.2 21 Ester value 168.0-l 75.0 2, 13, 14 (sap. val.) (acid val.) Iodine value 1 .O maximum 2, 13, 14, 20 2.4 (Wijs) 21 Moisture 0.8% 21 Ester content 99% 21 Unsaponifiable matter 0.5% maximum 21 Flash point: Closed cup 163.9T 21 160°C 10, 11, 25 Open cup 196°C 11 188.9”C 21

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FINAL REPORT:SAFETY ASSESSMENT OF STEARATES 113

TABLE 2. (Continued).

ingredient Reference

Autoignition 355T 25 temperature Residue on ignition 0.1% maximum 20 Weight/gallon 7.14 lb (2OT) 10 Dipole moment 1.88 D in benzene (24°C) 21 Cubical expansion 0.00083/T over 20-5OT 21 coefficient Vapor pressure 11 mm Hg (15OT) 21 Surface tension 31 dyne/cm (25°C) 21 Electrical conductivity 21 x 10-S (3OT) 21 Dielectric constant 3.111 (30°C) 21 Heat of fusion 39/40 Cal/g 21 Hydrolysis 0.16% (1 hr at 100°C) 21

lsobutyl Stearate

Molecular formula CA-LOI 8, 11 Molecular weight 340.57 11 Melting point 2OT 11 Saponification value 170-180 2 Acid value 1 .O maximum 2 Iodine value 1 .O maximum 2

lsocetyl Stearate

Molecular formula C,.HsaOz 8 Molecular weight 508 - Specific gravity 0.852-0.858 (25O/25”C) 17, 23 Freezing point O.lOOC 17, 23 Viscosity 32.0 cp (25T) 10 Refractive index 1.4510-l .4530 (25“C) 17, 23 Saponification value 110-118 1 106-l 18 17 Acid value 2.0 maximum 1 1 .O maximum 17 Iodine value 2.0 maximum 1 1 .O maximum 17

Isopropyl Stearate

Molecular formula Cd-LO, 8 Molecular weight 326 - Ester value 176-185 1 Acid value 1 .O maximum 1 Iodine value 1 .O maximum 1

Octyl Stearate

Molecular formula C&LO~ 8 Molecular weight 396 - Ester value 144-154 1 Acid value 1 .O maximum 1 Iodine value 1 .O maximum 1

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114 COSMETIC INGREDIENT REVIEW

Reactivity

The Stearates can be expected to undergo chemical reactions typical of esters.(241 Such reactions may include:

1. Conversion into stearic acid and the corresponding alcohol by chemical or enzymatic hydrolysis 2. Conversion into amides by ammonolysis 3. Conversion into different esters by alcoholysis or transesterification.

The purer grades of these saturated stearate compounds would not be expected to autoxidize readily.

Compatibility with Other Materials

Butyl Stearate is reported to be “compatible” with a number of materials, including butadiene/styrene rubber, butadienelacrylonitrile rubber, benzyl cellulose, copal esters, cellulose nitrate, cellulose acetate butyrate, cellulose acetate propionate, chlorinated rubber, ester gums, ethyl cellulose, nitrocellulose, polystyrene, and phenolic resins.(2”

Analytical Methods

Among the analytical methods that have been used for the various Stearates are the following: gas chromatography, (26-34) thin-layer chromatography,‘35-37’ silver resin chromatography,‘38~3g’ silver nitrate high performance liquid chromatography, (40) and photodensitometry.‘42’

USES

Noncosmetic Uses

Butyl Stearate is used as a solvent for dyes in wax polishes, as a spreading and softening agent in plastics, textiles and rubber, as a plasticizer for inks, cellulose acetate butyrate, cellulose nitrate, ethyl cellulose, polystyrene, laminated fiber products, rubber hydrochloride, chlorinated rubber and cable lacquers, as a lubricant in textiles, molding, metals, and in the extrusion and molding of polyvinyl- chloride, as a waterproofing agent for concrete, as an emollient in pharmaceuticals, and as an ingredient of carbon paper, inks, and special lubricants and coatings. (‘J’.‘~) The numerous and varied uses of Butyl Stearate have been exten- sively reviewed by Lower.(21) A number of regulations pertaining.to the use of Butyl Stearate as an indirect food additive, direct food additive, and synthetic flavoring agent have been issued by the Food and Drug Administration. As an indirect food additive, Butyl Stearate is permitted for use as a component of adhesives used in packaging or transporting food,““) as a plasticizer of resinous and polymeric coatings,(44’ as a plasticizer in resinous and polymeric coatings for polyolefin films,(45) as a component of defoaming agents used in coatings,(46) as a plasticizer in rubber articles

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FINAL REPORT:SAFETY ASSESSMENT OF STEARATES 115

(not to exceed 30 percent by weight of rubber product),(47) as a surface lubricant used in the manufacture of metallic articles,(48) and as a plasticizer in the manufacture of various other food packaging materials. (4g)As a direct additive, it is permitted as a defoaming agent component (no limitations) in the processing of beet sugar and yeast. (‘O)As a synthetic flavor, it may be safely incorporated into foods when used in accordance with good manufacturing practices and when used in the minimum quantity required to produce its intended effect.(51) Butyl Stearate is reported to be used in various beverages and foods in the following quantities: (12)

Nonalcoholic beverages 1 .O ppm Alcohol beverages 5.0 ppm Ice creams, ices, and so on 2.0 ppm Candy 190 ppm Chewing gum 330 ppm Baked goods 340 ppm

lsobutyl Stearate is used as a component in waterproof coatings, polishes, ointments, dye solutions, inks, lubricants, and rubbers.“‘) lsocetyl Stearate is used as a plasticizer, mold release agent, and textile softener. In pharmaceuticals, the isocetyl ester is used as a lubricant, fixative, and solvent.(“)

Cosmetic Use

The Stearate ingredients are used in cosmetics primarily as emollients for the skin.(g) In lipstick formulations, Butyl Stearate reduces the viscosity of the oil phase, thereby lessening the drag of the stick on the lips. Butyl Stearate may also function in lipsticks as a color-suspending agent. The low viscosity and oily nature of the compound allow it to wet and dissolve pigments more efficiently than oils of greater viscosity.‘8~‘0~“~‘8~2’~52’I n nail varnishes, Butyl Stearate may be used as a water-repelling plasticizer. In hand creams and lotions, Butyl and Iso- propyl Stearate function as spreading agents. When these 2 compounds are applied to the skin, a thin, oily film is deposited. The film is nongreasy and nontacky and is both continuous and hydrophobic in nature. Isopropyl Stearate also serves to increase the gelatin characteristics of hand product formulations.‘18,21) lsocetyl Stearate is used in cosmetics as a lubricant, fixative, and solvent.(10) Data submitted to the Food and Drug Administration (FDA) in 1981 by cosmetic firms participating in the voluntary cosmetic registration program indicated that Butyl, Cetyl, Isocetyl, Isopropyl, Myristyl, and Octyl Stearate were used in a total of 116, 4, 58, 16, 1, and 10 cosmetic formulations, respectively (Table 3). Product types in which the Stearate esters were most frequently used included eye makeup preparations, skin makeup preparations, lipstick, and skin care preparations. Concentrations of the Stearate esters in cosmetic products generally ranged from 1 to 25 percent, although there were a few reported instances of higher and lower concentrations. (53,54)No product formulation data were available for lsobutyl Stearate. Voluntary filing of product formulation data with the FDA by cosmetic manufacturers and formulators conforms to the prescribed format of preset concentra-

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116 COSMETIC INGREDIENT REVIEW

TABLE 3. Product Formulation Data’S3.s”

No. of Product formulations Within Total No. of Total No. Each Concentration Range (%J Formulations Containing Product Category in Category fngredient > JO-25 >5-10 >J-5 >O.l-I

Butyl Stearate

Eyeliner 396 4 4 Eye shadow 2582 3 - - 2 Other eye makeup preparations 230 4 1 3 - Hair conditioners 478 3 - 1 2 Makeup foundations 740 1 1 - - Lipstick 3319 78 69 1 8 Nail polish and enamel remover 41 2 - 1 Deodorants (underarm) 239 1 - 1 Feminine hygiene deodorants 21 1 - - Preshave lotions (all types) 29 3 2 1 Skin cleansing preparations 680 1 - 1 (cold creams, lotions, liquids, and pads) Face, body, and hand skin care 832 8 - 6 preparations (excluding shaving preparations) Moisturizing skin care prep 747 2 - 2 arations Wrinkle smoothers (removers) 38 1 - - Other skin care preparations 349 2 1 1 Suntan gels, creams, and liquids 164 2 2 -

198 1 TOTALS 116 74 29

Cetyl Stearate

Eye shadow 2582 3 - 3 - Face powders 555 - - 1

1981 TOTALS 4 3 1

No. of Product Formulations Within Each Total No. of Total No. Concentration Range (96) Formulations Containing Product Category in Category ingredient > TO-25 >5-10 >l-5 >o. I-1 so. 1

lsocetyl Stearate

Eye shadow 2582 1 - 1 - - - Eye makeup remover 81 1 - - - - Blushers (all types) 819 - 1 1 - - Face powders 555 1 - 1 - - Makeup foundations 740 6 1 - - - Lipstick 3319 - - 1 - - Makeup bases 831 7 - - 1 - Rouges 211 1 - - - - Nail creams and lotions 25 - - 1 - - Bath soaps and detergents 148 - - - - 1 Other shaving preparation 29 - - 1 - - products

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TABLE 3. (Continued)

NO. of Product Formulations Within Each Total No. of Total No. Concentration Range (%) Formulations Containing Product Category in Category fngredient > IO-25 >5-10 >I-5 >O.l-1 SO.1

Skin cleansing prepara- 680 5 - - 2 - 3 tions (cold creams, lotions, liquids, and pads) Face, body, and hand 832 11 - - 10 1 - skin care preparations (excluding shaving preparations) Moisturizing skin care 747 10 - - 9 - 1 preparations Night skin care prepara- 219 2 - 1 - 1 - tions Other skin care prepara- 349 2 - 1 - 1 - tions Suntan gels, creams, and 164 1 1 - - - - liquids Other suntan preparations 28 1 - - - 1 -

1981 TOTALS 58 17 5 26 5 5

No. of Product Formulations Within Each Total No. of Total No. Concentration Range (%) Formulations Containing Product Category in Category Ingredient >25-50 > IO-25 >5-10 >l-5 >O.l-I

isopropyl Stearate

Bath oils, tablets, and salts 237 1 - - 1 - - Eye makeup preparations 230 1 - 1 - - - Fragrance preparations 191 1 1 - - - - Personal cleanliness 227 3 - - - 3 - products Skin cleansing prepara- 680 4 - 1 3 - - tions (cold creams, lotions, liquids, and pads) Moisturizing skin care 747 4 - 2 2 preparations Night skin care prepara- 219 1 - 1 - - - tions Skin lighteners 44 1 - 1 -

1981 TOTALS 16 1 6 6 3 -

Makeup foundations 740 - - - 1 -

198 1 TOTALS - - - 1 -

Octyl Stearate

Bath oils, tablets, and salts 237 3 - - - 2 1

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TABLE 3. (Continued)

No. of Product Formulations Within Each Total No. of Total No. Concentration Range (%) Formulations Containing Product Category in Category Ingredient >25-50 > IO-25 >5-10 > l-5 >O.l-I

Colognes and toilet 1120 4 - - - - 4 waters Makeup foundations 740 1 - - - - 1 Face, body, and hand 832 1 - - - - 1 skin care preparations (excluding shaving preparations) Night skin care prepara- 219 1 - 1 - - 1 tions

1981 TOTALS 10 - 1 - 2 7

tion ranges and product categories as described in Title 21 Part 720.4 of the Code of Federal Regulations. cs5)Because data are only submitted within the framework of preset concentration ranges, opportunity exists for overestimation of the actual concentration of an ingredient in a particular product. An entry at the lowest end of a concentration range is considered the same as one entered at the highest end of that range, thus introducing the possibility of a 2- to lo-fold error in the assumed ingredient concentration. Cosmetic products containing the Stearate compounds are applied to or have the potential to come in contact with eyes, hair (scalp), nails, vaginal mucosa, and skin. Small amounts of these esters could be ingested from lipstick. Product formulations containing the Stearate ingredients may be used as infrequently as once a week to as frequently as several times per day. Many of these cosmetic products can be expected to remain in contact with body surfaces for as briefly as a few hours to as long as a few days. Each cosmetic product containing one of.these esters can be repeatedly applied over the course of many years.

METABOLISM

Higher molecular weight aliphatic esters are readily hydrolyzed to the corresponding alcohol and acid and then generally oxidized to carbon dioxide and water.“‘) In the case of the Stearate compounds, metabolism of stearic acid would be expected to occur in the same fashion as other fatty acids. Isopropyl Stearate diluted in 9,10-3H,-labeled oleic acid was given by stomach tube to thoracic duct fistula rats. Measurement data of the specific radioactivity of the stearic chains in lymph triglycerides indicated that 85 to 95 percent of the chains were of “dietary origin.” For every 100 radioactive stearic chains recovered in lymph lipids, less than 10 were in the form of isopropyl ester, which represented 2 to 3 percent of the total lymph lipids. It was calculated that less than 5 percent of the remaining radioactivity was in lymph phospholipids. Of the non-

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phospholipid fraction (isopropyl esters being excluded), 95 percent or more of the radioactivity was detected in the triglycerides. It was concluded that “. . .a part of the isopropyl ester was hydrolyzed in the intestine, and that the acids thus liberated were reesterified and partitioned between lymph lipids.“‘56’

ANIMAL TOXICOLOGY

Comedogenicity

Butyl Stearate (50 percent in mineral oil) and mineral oil were each applied to 1 ear of 2 rabbits 5 days a week for 2 weeks. The treated sites were excised at 14 days, and comedogenicity was graded on a scale of 0 to 3. For 50 percent Butyl Stearate in mineral oil, scores were 2 and 3, indicating moderate and strong comedogenicity, respectively. Mineral oil produced reactions of grade 1 in both rabbits, indicating slight comedogenicity. These materials were only weakly comedogenic in relation to potent acnegens, such as tars and chlorinated oils. The authors discussed the possibility that daily use of Butyl Stearate for several years could produce low-grade comedonal reactions in susceptible women.(“) The CIR Panel recognizes that currently available tests are inadequate to predict the potential for human comedogenicity of a cosmetic ingredient as used in a product formulation. However, comedogenicity is a potential health effect that should be considered when the stearate ingredients are used in cosmetic formulations.

Subcutaneous Toxicity

Undiluted Butyl Stearate was given by subcutaneous injection to 4 groups of male albino rats consisting of 6, 15, 6, and 15 animals at single doses of 4, 8, 16, and 32 g/kg, respectively. No deaths or gross lesions were observed in any of the animals.‘58’

lntraperitoneal Toxicity

Four groups of male albino rats were administered undiluted Butyl Stearate by intraperitoneal injection. Single doses of 4, 8, 16, and 32 g/kg were administered to 6, 15, 6, and 15 animals, respectively. No gross lesions or deaths were noted.‘“*)

Acute Oral Toxicity

Results of studies conducted on rats and mice have indicated that Butyl, Iso- cetyl, Isopropyl, Myristyl, and Octyl Stearates have low acute oral toxicity. The individual acute oral studies are discussed below; results are summarized in Table 4. Undiluted Butyl Stearate was administered in single oral doses of 4, 8, 16, and 32 g/kg to 4 groups of male albino rats consisting of 6, 15, 6, and 15 animals, respectively. No deaths or gross lesions were noted in any of the animals. The acute lethal oral dose was >32 g/kg.(58)

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TABLE 4. Acute oral Toxicity

s Concentration No. and Kind ingredient I%) Dose of Animal LDra Comments Reference

Butyl Stearate 100 4 to 32 g/kg 6, 15, 6, and >32 g/kg No deaths or gross pathologic changes 58 15 albino rats, respectively Butyl Stearate Each sample: - Rats ‘5 g/kg 9, 59 (2 samples) 100 Butyl Stearate 100 10 g/kg 10 rats >lO g/kg No deaths 9, 60 Butyl Stearate 100 8 ml/kg 5M and 5F al- >8 ml/kg No deaths; average body weight gain was 9, 61 bino rats 21.6% over 14-day observation period Butyl Stearate 100 2.0-64.0 6 groups of 5 > 64.0 ml/kg No effect at 2.0 ml/kg. Rats given 4.0 and 9, 62 ml/kg albino rats 8.0 ml/kg had unkempt coats. Rats given 16.0 and 32.0 ml/kg developed diarrhea and oily, unkempt coats. Rats given 64.0 ml/kg had diarrhea, alopecia, and oily, unkempt coats. Test material equally nontoxic to males and females lsocetvl Stearate 100 0.464-10.0 5 groups of 5M > 10.0 ml/kg Rats given 0.464, 1 .O, and 2.15 ml/kg ex- 9, 63 ml/kg albino rats hibited normal appearance and behavior. Rats given 4.64 ml/kg had unkempt fur. At 10.0 ml/kg, depression and unkempt fur were noted. No deaths or gross pathological alterations observed at any dose level 9, 9, 64 8 lsocetyl Stearate 100 5 g/kg 5M and 5F al- > 5.0 g/kg One female rat died within 24 hours; bino rats gross necropsy revealed fibrous tissue z encasing heart and lungs of 1 male =! lsocetyl Stearate 100 10 g/kg 10 rats >lO g/kg No deaths 9, 65 n Isopropyl Stearate 100 8 ml/kg 5M and 5F al- >8 ml/kg l/10 rats died. Body weight gain aver- 9, 66 z bino rats aged 21% over the 2-week observation 5 5 period Myristyl Stearate 100 mg/ml of 5.0 or 10.0 2 groups of 5 > 10 g/kg (ingred. + No deaths or visible “untoward effects” 9, 67 ! corn oil glkg and 20 CFW corn oil); >l g/kg 5 5 mice, respec- (ingred. alone) tively E 9 , 9, 68 < Octyl Stearate 100 8 ml/kg 5M and 5F rats >8 ml/kg No deaths; body weight gain averaged ;; 25.7% during the 2-week observation s period

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FINAL REPORT:SAFETY ASSESSMENT OF STEARATES 121

Two samples of 100 percent Butyl Stearate were tested for acute oral toxicity in an unspecified number of rats. The LDso of each sample was >5 g/kg.‘9s59’ A single 10 g/kg dose of undiluted Butyl Stearate was given orally to 10 rats. No deaths occurred during the 72-hour observation period. The investigators concluded that the LDso was >lO gIkg.‘9*60’ Five male and five female albino rats were administered 100 percent Butyl Stearate at an oral dose of 8 ml/kg. No rats died during the 1Qday observation period. The LDsO was >8 ml/kg.‘9*61) Doses of 2.0 to 64.0 ml/kg of undiluted Butyl Stearate were given orally to 6 groups of albino rats (5 animals/group). Rats dosed at 2.0 ml/kg had no adverse effects. Unkempt coats were observed for 12 to 48 hours in animals receiving 4.0 and 8.0 ml/kg. Rats administered 16.0 and 32.0 ml/kg had slight diarrhea and wet, oily, unkempt coats; these animals returned to normal within 3 to 6 days. Slight to moderate diarrhea, alopecia, and wet, oily, unkempt coats were noted in rats dosed at 64.0 ml/kg. However, these animals appeared normal within 7 to 8 days. The test material was “equally nontoxic” to males and females. The LDsO of 100 percent Butyl Stearate was >64.0 ml/kg.(9,69) Undiluted lsocetyl Stearate was given orally to 5 groups of 5 male Sprague Dawley rats at doses ranging from 0.464 to 10.0 ml/kg. All of the rats of the 0.464, 1 .OO, and 2.15 ml/kg groups showed normal appearance and behavior throughout the 1Cday observation period. Rats administered 4.64 ml/kg had normal appearance and behavior during the day of dosage and from Days 4 through 14. However, on the first, second, and third day postdosing, unkempt fur was noted. At the 10.0 ml/kg dose, “depression” was observed on the day of dosing and unkempt fur observed on Days 1 through 3. From Day 4 through 14, all animals had normal appearance and behavior. Necropsy was performed on the rats of each dose group, and no gross alterations were observed. The average body weight gain for each group was normal for rats of the age, sex, and strain used in this study. No deaths occurred at any dose.(9*63) Five male and five female albino rats received undiluted lsocetyl Stearate at an oral dose of 5.0 g/kg. One female rat died during the first 24 hours. Necropsy was conducted on each of the animals at the end of the 14-day observation period, and fibrous tissues encasing the heart and lungs were found in 1 male. The investigators considered the test material nontoxic under conditions of this test (9.64) A single 10 g/kg oral dose of undiluted lsocetyl Stearate was administered to 10 rats. No deaths occurred during the 72-hour observation period. The investigators considered the test material nontoxic.(9*65) Five male and five female albino rats were dosed orally with 8 ml/kg of undiluted Isopropyl Stearate. On the eighth day of the 14day observation period, 1 of the 10 rats died; necropsy findings were noncontributory as to the cause of death. The body weight gain of the test animals averaged 21 percent over the 2- week observation period. Under conditions of this test, the LDso was >8 ml/ (9.66) kg. CFW mice of the Carworth strain were administered Myristyl Stearate in corn oil (100 mg/ml) at oral doses of 5.0 (5 mice) or 10.0 g/kg (20 mice). During the 5-day observation period, no deaths or visible “untoward effects” were observed. (9,67)

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122 COSMETIC INGREDIENT REVIEW

Undiluted Octyl Stearate was administered to 5 male and 5 female rats at oral doses of 8.0 ml/kg. No deaths occurred during the 14day observation period. The body weight gain of test animals during the 2 weeks averaged 25.7 percent. The investigators considered the acute oral toxicity of this ester to be “very low. fr(9.68)

Eye Irritation

When instilled into the eyes of rabbits at concentrations of 100 percent, Iso- cetyl, Myristyl, and Octyl Stearates caused slight, transient irritation. No ocular irritation was observed in rabbits following instillation of 100 percent Butyl or Iso- propyl Stearate. The various eye irritation studies are individually discussed below; results are summarized in Table 5. Three groups of albino rabbits (3 animals/group) were treated with 100 percent Butyl Stearate for ocular irritation. The test material (0.1 ml) was instilled into the right eye of each animal; the untreated left eye served as control. The eyes of the first group of rabbits received no further treatment. In the second group, the treated eyes were washed with 20 ml of lukewarm water 2 seconds after instillation of the test material. In the third group, the treated eyes were rinsed in a similar fashion 4 seconds after instillation of the test substance. Both the treated and control eyes were examined every 24 hours for 4 days and then again on the seventh day. No eye irritation was observed in any of the animals.(9’62) Two groups of rabbits (3 animals/group) were treated with 100 percent Iso- cetyl Stearate for ocular irritation. The eyes of the first 3 rabbits did not receive a water rinse after instillation of 0.1 ml of the test material. The eyes of the second group of rabbits were flushed with 20 ml of water 4 seconds after instillation of 0.1 ml lsocetyl Stearate. The average scores of the “no-wash” group were 7.33, 1.33, and 0.0 at the 1, 24, and 48-hour readings, respectively (maximum score, 110). The average scores of the rabbits receiving the 4-second “wash” were 10.67, 2.0, and 0.0 at the 1, 24, and 48-hour readings, respectively. According to the investigators, lsocetyl Stearate was slightly irritating.(9.70) An eye irritation study was conducted with undiluted lsocetyl Stearate using 6 New Zealand rabbits. The test substance (0.1 ml) was instilled into the right eye of each animal; the untreated left eye of each rabbit served as a control. The treated eyes of all rabbits were not flushed with water. Numerical scores for ocular lesions were assigned according to the Draize(‘l) scoring system of 110 maximum points. At the 24hour reading, 4 of the 6 animals had varying degrees of redness of the palpebral conjunctiva. Of the 4 rabbits with irritation, only 1 had chemosis. The 24-hour Draize scores for the 6 rabbits were 0, 0, 2, 2, 2, and 8, respectively. On the second day, all treated eyes were normal and remained so until the final observation on Day 3. The investigators concluded that the test material was a minimal ocular irritant to rabbit eyes under conditions of this test.(9,64) Undiluted Isopropyl Stearate was instilled into the conjunctival sac of one eye of each of 6 New Zealand rabbits (3M, 3F). The treated eye received no further treatment. The single 0.1 ml exposure produced no corneal, conjunctival, or iridic irritation over a 3-day observation period.(“) The eye irritation potential of undiluted Myristyl Stearate was determined in 3 albino rabbits. Each animal had 0.1 g of the test sample instilled into the right

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TABLE 5. Eye Irritation

No. Concen- of tration Rab- Wash (WY ingredient Pw bits No Wash (NW) Comments/Results Reference

Butyl 100 NW No irritation observed on Days 1, 2, 3, 4, or 7 9, 62 Stearate W after 2 sec- No irritation observed on Days 1, 2, 3, 4, or 7 onds W after 4 sec- No irritation observed on Days 1, 2, 3, 4, or 7 onds lsocetyl 100 NW The average scores were 7.33, 1.33, and 0.0 9, 70 Stearate at the 1, 24, and 48 hour readings, respectively. (The max. score per observation interval = 110) 3 W after 4 sec- The average scores were 10.67, 2.0, and 0.0 onds at the 1, 24, and 48 hour readings, respectively. (The max. score per observation interval = 110.) The investigators considered the material a slight irritant lsocetyl 100 6 NW At 24-hour reading, 4/6 rabbits had redness of 9, 64, 71 Stearate conjunctivae. The 24-hour Draize scores of the 6 rabbits were 0, 0, 2, 2, 2, and 8, respectively. (The max. score per rabbit observation = 110.) No irritation observed on Days 2 or 3. Investigators considered the material a minimal irritant

Isopropyl 100 6 NW No corneal, conjunctival, or iridic irritation 72 Stearate noted over a 3-day observation period

Myristyl 100 3 NW At the 24.hour reading, all 3 rabbits had a 9, 73 Stearate “slight vessel injection involving only the conjunctivae.” The 24-hour Draize scores for the 3 rabbits were 2, 4, and 6, respectively. (The max. score per rabbit per observation = 110.) No irritation was observed on Days 2, 3, 4, or 7

Octyl 100 6 NW Average ocular irritation indices at 1 hour and 74, 75 Stearate 1, 2, 3, 4, and 7 days were 4.67, 0, 0, 0, 0, and 0, respectively. (The max. score per observation period = 100.) Material did not provoke any significant eye injury

eye with no further treatment. The left eye of each rabbit served as a control. Both treated and control eyes were examined every 24 hours for 4 days and then again on the seventh day. Eye irritation was evaluated according to the Draize scale of 110 maximum points per rabbit per observation. At the 2.4hour reading, all 3 rabbits had a “slight vessel injection” involving only the conjunctiva. On the second day of observation, the treated eyes were normal and remained so until the final evaluation. The 24-hour Draize scores for the 3 rabbits were 2, 4, and 6, respectively. (9.73)

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Undiluted Octyl Stearate was tested for eye irritation in 6 albino rabbits by methods outlined in the journal Officiel de /a Republique Francaise. (75) The test material (0.1 ml) was instilled into 1 eye of each animal; the other eye served as a control. The treated eyes of all animals received no water rinse. Ocular lesions were evaluated on the basis of a numerical scale ranging from 0 to 100. The average ocular irritation indices at 1 hour and at 1, 2, 3, 4, and 7 days were 4.67,0, 0, 0, 0, and 0, respectively. According to the investigators, “. . .a compound does not provoke any significant injury to the eye mucous membrane when. . .the ocular index is less than 10.“(74)

Skin Irritation

When tested at concentrations of up to 100 percent on rabbit skin, Butyl, Iso- butyl, Isocetyl, Isopropyl, Myristyl, and Octyl Stearates caused at most mild to moderate irritation. Individual tests are discussed below; results are summarized in Table 6. A skin irritation test was conducted with undiluted Butyl Stearate using 6 rabbits. The test material was applied to the abraded and intact skin under closed patches for 24 hours. All animals had slight to well-defined erythema at abraded and intact skin sites at the 24-hour evaluation. Two of the six rabbits had very slight edema at abraded sites at 24 hours. No skin irritation was observed in any of the animals at the 72-hour evaluation. The primary irritation index (PII) was 0.68, indicating mild irritation.C9n76’ Undiluted Gutyl Stearate was tested for skin irritation in 6 rabbits. The ester was applied to the intact or abraded skin under closed patches for 24 hours. Very slight erythema was noted at both abraded and intact skin sites of 2 rabbits at the 24-hour reading. No irritation was observed in any of the animals at the 72-hour reading. The PII was 0.17, indicating mild irritation.(9*77) The skin irritation potential of undiluted Butyl Stearate was determined in rabbits in 4 separate tests. In each test, the material was applied to the skin under closed patches for 24 hours. The Plls were 0.33 and 0.39 in 2 of the tests, respectively, indicating minimal irritation. (9v78)The Plls were 0.3 and 0.77 in the other 2 tests, indicating minimal to mild irritation. (9s9) Details of these tests were not reported. Undiluted Butyl Stearate was tested for its skin irritating effects by means of the procedures described in the Department of Transportation Act.(79) The test material was applied to the intact and abraded skin of 6 albino rabbits under closed patches for 4 hours. Following an initial evaluation at 4 hours, all test sites were washed with an “appropriate” solvent to prevent further exposure. No irritation was noted in any of the animals at the 4-, 24-, or 48-hour readings; the PII was 0.0, indicating that the test material was “not a primary irritant.“(9,69’ The Draize methodrEO) was used to evaluate the skin-irritating effects of undiluted Butyl Stearate. The test material (0.5 ml) was applied for 24 hours under closed patches to the abraded and intact skin of 6 albino rabbits. No irritation was noted in any of the animals at 24 or 72 hours; the PII was 0.0, indicating no observed skin irritation.(9*69)

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FINAL REPORT:SAFETY ASSESSMENT OF STEARATES 125

Six rabbits were treated with 100 percent Butyl Stearate for evaluation of primary skin irritation. The test procedures employed were those as specified in the regulations under the Federal Hazardous Substances Act.@l’ The test material was applied to both abraded and intact skin under occlusive patches for 24 hours. No irritation was noted in any of the rabbits at the 24- or 72-hour readings; the PII was 0.0.C9*82) Undiluted Butyl Stearate was tested for skin irritation using 6 albino rabbits. The test material was applied under occlusive patches to the intact and abraded skin for 24 hours. At the 24hour evaluation, all animals had well-defined to moderate erythema at both intact and abraded skin sites. Further, 2 of 6 rabbits had very slight edema of the intact skin and 1 of 6 rabbits had very slight edema of the abraded skin. By the 72-hour reading, all animals had well-defined to severe erythema but no edema. The PII was 2.75, indicating moderate irritation.(9,83) A skin irritation test was conducted with undiluted lsobutyl Stearate on 6 rabbits. The test material was applied to the intact and abraded skin under occlusive patches for 24 hours. At the 24-hour evaluation, all 6 animals had very slight erythema of the intact skin, whereas 5 of the 6 rabbits had very slight erythema of the abraded skin. At the 72-hour evaluation, 2 of the 6 rabbits had very slight erythema at both abraded and intact skin sites. The PII was 0.62, indicating mild irritation. (9*84) The skin irritation potential of undiluted lsocetyl Stearate was determined using 6 New Zealand rabbits. The test material was applied to the abraded and intact skin of both the back and abdomen under closed patches for 24 hours. The irritant effects following dorsal application were confined to very slight erythema at 1 of 6 intact and 2 of 6 abraded sites at the 24hour evaluation; the PII was 0.13, indicating mild irritation. The irritant effects observed following ventral application included very slight erythema at 2 of 6 intact sites and 1 of 6 abraded sites at the 24hour evaluation. At the 72-hour evaluation, very slight erythema was noted at 1 of 6 intact sites and 1 of 6 abraded sites. The PII of the test material was 0.21, indicating mild irritation.(9.63) Two samples (A and B) of undiluted lsocetyl Stearate were tested for skin irritation on an unspecified number of rabbits. The test materials were applied to the skin daily for 3 days; “open patch contact time” was 24 hours. The PII scores for sample A were 0.17, 1 .OO, and 0.83, whereas sample B had PII scores of 0.00, 0.83, and 0.83. The investigators concluded that the 2 samples were “slight” skin irritants.(9,85) An unspecified number of rabbits were used for skin irritation tests with 2 samples (A and B) of undiluted lsocetyl Stearate. The test materials were applied to the skin under closed patches for 24 hours. Sample A had a PII of 0.25, whereas sample B produced a PII of 0.12 (maximum score, 8.0). The investigators concluded that the 2 samples were “minimal” skin irritants.(9s85’ Six rabbits were treated with undiluted lsocetyl Stearate to determine its skin irritation potential. The test material was applied to the abraded and intact skin of the back under gauze pads for 24 hours. Within 24 hours, 3 of 6 animals had very slight erythema or edema of the intact and abraded skin sites. At 72 hours, 1 of the 6 rabbits had very slight erythema of the intact skin, and another had a similar reaction on abraded skin. The PII was 0.415, indicating mild irritation.(9*86)

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TABLE 6. Skin Irritation

Concentration No. of ingredient Pw Rabbits Method Comments/Results Reference

Butyl Stearate 100 6 Material applied to intact and abraded PII,* 0.68; mild irritation 9, 76 skin for 24 hours; closed patches

Butyl Stearate 100 6 Material applied to intact and abraded PII, 0.17; mild irritation 9, 77 skin for 24 hours; closed patches

Butyl Stearate 100 Unspecified In 4 separate tests, material was ap- In 4 separate tests, the Plls were 0.33, 9, 59, 78 plied under closed patches for 24 0.39, 0.3, and 0.77, respectively, hours indicating minimal to mild irritation

Butyl Stearate 100 6 Dept. of Transportation. VW Material PII, 0.0; no irritation observed 9, 69 applied to intact and abraded skin for 4 hours; closed patches

Butyl Stearate 100 Draize.‘*“’ Material applied to intact WI, 0.0; no irritation observed 9, 69 and abraded skin for 24 hours; closed patches

Butyl Stearate 100 Federal Hazardous Substances Act.‘*” PII, 0.0; no irritation observed 9, 82 Material applied to intact and abraded skin for 24 hours; closed patches

Butyl Stearate 100 Material applied to intact and abraded PII, 2.75; moderate irritation 9, 83 skin for 24 hours; closed patches lsobutyl Stearate 100 Material applied to intact and abraded PII, 0.62; mild irritation 9, 84 skin for 24 hours; closed patches lsocetyl Stearate 100 Material applied to intact and abraded Back: PII, 0.13; mild irritation 9, 63 skin of both the back and abdomen Abdomen: PII, 0.21; mild irritation for 24 hours; closed patches lsocetyl Stearate Sample A 100 Unspecified Each sample applied to skin daily for A: Plls, 0.17, 1 .OO and 0.83; slight ir- 9. 85 3 days; “open patch contact time” ritation was 24 hours

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Sample B 100 Unspecified B: PBS, 0.00, 0.83, and 0.83; slight irritation lsocetyl Stearate Sample A 100 Unspecified Each sample applied to skin for 24 A: PII, 0.25; minimal irritation 9, a5 hours; closed patches Sample B 100 Unspecified B: PII, 0.12; minimal irritation 9, 86 > lsocetyl Stearate 100 6 Material applied to intact and abraded PII, 0.415; mild irritation w skin for 24 hours; closed patches 2 lsocetyl Stearate 25 in corn oil 6 Draize et al. I”) Material applied to in- PII, 0.25; “potential for slight irrita- 9, 64 z tact and abraded skin for 24 hours; tion” 2 closed patches -4

Isopropyl Stearate 100 6 A single 0.5 ml application of the test PII, 2.35; moderate irritation 9, 83 8 material to intact and abraded skin Y Isopropyl Stearate 100 6 A single 0.5 ml application of the test PII, 2.35; moderate irritation 87 F material to intact and abraded skin 9, 88 5 Myristyl Stearate 100 3 Draize.r80’ Material applied to intact PII, 0.0; no irritation observed Fl and abraded skin for 24 hours; closed patches

Octyl Stearate 100 6 /ournal Officiel de la Republique Fran- PII, 0.0; no irritation observed 74 caise. C’S) Material applied to skin for 24 hours; closed patches

Octyl Stearate 10 in aqueous 6 journal Officiel de la Republrque Fran- PII, 0.0; no irritation observed 74 solution cake. ‘75r Material applied to skin for 24 hours; closed patches

Octyl Stearate 100 6 Federal Hazardous Substances Act.“” PII, 1 .42; mild irritation 9, a9 Material applied to intact and abraded skin for 24 hours; closed patches - *PII, primary irritation index.

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The Draize method”‘) was used to evaluate the skin irritation potential of 25 percent lsocetyl Stearate in corn oil. The test material was applied to the intact and abraded skin of 6 albino rabbits for 24 hours under occlusive wrapping. Very slight erythema was observed at both intact and abraded skin of 3 rabbits at the 24-hour evaluation; no signs of irritation were noted in any animals at the 72- hour evaluation. The PII was 0.25, indicating a potential for slight irritation.(9.64) Albino rabbits were treated with undiluted Isopropyl Stearate for evaluation of primary skin irritation. The test material was applied under occlusive patches to the intact and abraded skin of each of 6 animals for 24 hours. Slight to moderate erythema was observed in all animals at both intact and abraded skin sites at the 24- and 72-hour evaluations. The PII was 2.35, indicating moderate irritation (9.83) Well-defined erythema was observed at the intact and abraded skin sites of 6 rabbits (3M, 3F) both 24 and 72 hours following a single 0.5 ml application of undiluted Isopropyl Stearate. The PII of the test material was 2.35, indicating moderate skin irritation.“‘) The Draize procedure(80) was employed to determine the skin irritation potential of undiluted Myristyl Stearate. The test material was applied for 24 hours under closed patches to the intact and abraded skin of 3 albino rabbits. No irritation was noted in any of the animals at the 24- or 72-hour readings. The PII was 0.0, indicating no irritation.‘9,*8) The skin irritation potential of undiluted Octyl Stearate and an aqueous solution containing 10 percent Octyl Stearate was determined by procedures outlined in the journal Officiel de la Republique Francaise. (75) Each test material (0.5 ml) was applied under occlusive patches for 24 hours to the clipped skin of 6 albino rabbits. Test sites were scored at 24, 48, and 72 hours. The primary irritation index of both test materials was 0.0, indicating no irritation.(74) The skin-irritating effects of undiluted Octyl Stearate were determined according to the methods specified in the regulations under the Federal Hazardous Substances Act as cited in 16 CFR 1500.3 and 16 CFR 1500.41. The test material was applied under closed patches for 24 hours to the intact and abraded skin of 6 New Zealand rabbits. Well-defined erythema was noted at 24 hours in all rabbits on both intact and abraded sites; by 72 hours, the erythema had cleared on both sites in 5 of 6 animals. Very slight to slight edema was noted in 3 of 6 rabbits at 24 hours and 1 of 6 rabbits at 72 hours. The PII was 1.42, indicating mild irritation. However, the test material was not considered an irritant under the criteria outlined in Title 16 Part 1500.41 of the Code of Federal Regulations.(9,81.89)

Cumulative Skin Irritation

The cumulative skin irritation potential of 10 percent Octyl Stearate in aqueous solution and 100 percent Octyl Stearate was determined in albino rabbits by procedures outlined in the lournal Officiel de la Republique Francaise. (75) Each test material was applied to the shaved skin of 3 rabbits daily for 6 days. The rabbits were immobilized with a collar to prevent licking of the treated areas. The “mean maximum irritation index” of rabbits treated with undiluted Octyl Stearate was 0.67, indicating that it was “poorly tolerated.” Gross examination of the treated skin in all 3 rabbits revealed vesicles and slight epidermal exfoliation. Mi-

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FINAL REPORT:SAFETY ASSESSMENT OF STEARATES 129 croscopic changes included epidermal acanthosis and “congestive dermatitis.” The “mean maximum irritation index” of rabbits administered 10 percent Octyl Stearate in aqueous solution was 0.33, indicating that the material was “relatively well tolerated.” Grossly, vesicles were found in 2 rabbits, but no significant microscopic changes were observed.“‘)

Skin Sensitization

The skin sensitization potential of 0.1 percent Butyl Stearate in physiological saline was determined in 2 white male guinea pigs. The test material was injected “intracutaneously” every other day or 3 times weekly until a total of 10 injections had been made. The first injection consisted of 0.05 ml of the test material, whereas the subsequent 9 injections consisted of 0.1 ml each. Two weeks following the tenth injection, a challenge injection was made using 0.05 ml of a freshly prepared test solution. Twenty-four hours after each injection, evaluations were made of the diameter, height, and color of skin reactions. No skin sensitization was observed in either animal.(9,90) The skin sensitization potential of 0.1 percent lsocetyl Stearate in physiological saline was also determined in 2 white male guinea pigs. The test procedure employed was that as described above for Butyl Stearate. The saline solution containing 0.1 percent lsocetyl Stearate was nonsensitizing.(9*91)

Embryotoxicity and Effects on Reproduction

Groups of 20 male and 20 female rats were fed diets containing 6.25 percent Butyl Stearate for 10 weeks and then mated. No adverse effects were noted with respect to fertility, litter size, or survival of offspring. However, growth was significantly retarded during the preweaning and postweaning periods. No gross lesions were found among rats killed at the end of the 21-day postweaning period. (W

Chronic Toxicity

In a 2-year feeding study, concentrations of either 0.0, 1.25, or 6.25 percent Butyl Stearate were provided in the diet to 3 groups of 16 male rats. Daily doses of the test material corresponded approximately to 0, 2500, and 6000 mg/kg, respectively. No significant differences were observed between treated and control groups with respect to growth, survival, and hematological values. As the treated and control groups increased in age, slight changes in the ratio of types of leukocytes were observed. Differential counts of bone marrow smears at 12, 23, and 24 months indicated that ingestion of Butyl Stearate did not affect either cellular distribution or the myeloid:erythroid ratio. Gross lesions included acute and chronic inflammatory pulmonary changes, nephrosis, and fatty degeneration of the liver. Other pathologic changes, such as tumors and infections, were found primarily in older rats but were not related to the Butyl Stearate administration.

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130 COSMETIC INGREDIENT REVIEW

Histopathological alterations observed in treated and control groups included chronic pneumonitis, diffuse fatty infiltration of the liver, focal necrosis of hepatic cells surrounding veins, and chronic nephrosis.(58)

Carcinogenicity

No data were available on the carcinogenicity of any of the Stearates under review. However, data were available on methyl stearate and stearic acid. Female mice were injected subcutaneously with either 0.5 or 5.0 mg methyl stearate in 0.1 ml tricaprylin once a week for 26 weeks. Stearic acid was also tested in female mice at either 0.05 or 0.5 mg in 0.1 ml tricaprylin by subcutaneous injection once a week for a total of 26 injections. Mice receiving tricaprylin alone or no treatment served as controls. Results are presented in Table 7. In the 16 mice administered 5.0 mg methyl stearate, 2 subcutaneous and 2 pulmonary tumors were found. No tumors were observed in the stearic acid-treated mice. In the 16 vehicle control mice surviving at 6 months, a single breast carcinoma was observed. In the 171 untreated mice surviving 6 months, a total of 1 subcutaneous sarcoma, 10 pulmonary tumors, 14 mammary gland carcinomas, and 2 cutaneous carcinomas were recorded. The “background noise” of neoplasms in the mice used for the study was under 1 percent for subcutaneous sarcomas and under 10 percent for mammary and pulmonary tumors. Practically all tumors were found after 12 months of the study when the mice were 14 months old.(92) These data did not indicate carcinogenic activity for methyl stearate or stearic acid. In a study involving 2 separate laboratories, female mice were injected subcutaneously with either 0.5 or 5.0 mg methyl stearate in 0.1 tricaprylin once a week for 26 weeks. Stearic acid was tested at 0.05 or 0.5 mg/O.l ml of tricaprylin. Mice receiving tricaprylin alone or no treatment served as controls. The results of the 2 studies are summarized in Table 8. For methyl stearate, each laboratory observed 2 sarcomas at the site of injection after 21 months. According to the investigators, the findings with regard to methyl stearate were surprising, since the compound is relatively unreactive and its conversion in vivo to reactive species is difficult to envision. They considered the possibility that “solid-state” carcinogenesis was involved but noted that methyl stearate was lipid-soluble and gave clear solutions in the vehicle tricaprylin at the concentrations tested.(93) No tumors developed in any of 10 rats fed stearic acid as 0.3 percent of their diet for 209 days.(5.94) In a search for carrier materials for introducing potential carcinogens into mouse urinary bladders, stearic acid and other “inert vehicles” were tested for induction of bladder tumors. Pellets of stearic acid were implanted in the bladder of 62 mice and in those surviving 30 weeks. The bladder tumor incidence was 13 percent. The incidence of bladder tumors in mice implanted with either smooth (67 mice) or roughened (63 mice) glass beads was 4 and 29 percent, respectively. (95) Experiments involving a possible action of the vehicle or a physical effect of the agent, such as in studies by subcutaneous injection or bladder implantation, are included in this evaluation of Stearate ingredients. However, the results of such tests require careful consideration, particularly if they are the only ones rais- ing a suspicion of carcinogenicity.(96)

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TABLE 7. Test Results for Carcinogenic Activity’9” B .s’

Schedule Tumor by Month of Appearance sf Dose/ (Injection No./ Total dose Mice Alive at Month ;=I Injection * Week/ (mg Compound/ Mouse Subcutaneous Pulmonary Other 2 Compound (msJ Total Injections) ml Jricaprylin)t Strain* 0 6 9 12 15 18 Sarcomas Tumor Jumors§ * % Tricaprylin - l/26 012.6 s 16 16 14 14 - - - - B(l2) z

None - - - S 24 23 23 22 17 13 - None - - - 5 100 80 66 56 51 21 14 2414,17,17, B(1 1,14,- 5

17,17 15,17,17, 17,17,17, i l&l@, Y CU4,15) z None - - - S 47454313 8 3 - 18 B(11,13, F 16,18) 2 WI None - - - 5 32 23 23 14 13 - - 12,12,12 - Stearic Acid 0.5 1I26 1312.6 5 16 14 7 7 7 6 - - -

Stearic Acid 0.05 l/26 1.312.6 S 16 13 12 11 10 10 - - -

Methyl Stearate 5.0 1 I26 13012.6 S 16 16 16 16 14 11 15,15 15,15 -

Methvl Stearate 0.5 l/26 1312.6 s 16 16 16 15 15 - - - -

*Subcutaneous injection into inguinal area. +Tricaprylin used was >99% pure. $5, Swiss Webster female mice. §B, Breast carcinoma; C, cutaneous carcinoma.

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TABLE 8. Induction of Sarcomas at Injection Site by SC Injection in Mice. Results at 21 Monthsr93)

Dose (mg/O. I ml Jricaprylin) Total dose No. of Mice No. of Mice Sarcomas at Compound Laboratory Once Weekly* b-d at Start Alive at 6 Months+ injection Site Other Tumors

Methyl Stearate A* 5.0 130 15 15 2 1 0.5 13 15 15 0 3 B§ 5.0 130 16 16 2(15)‘1 2 0.5 13 16 16 0 0

Stearic Acid A 0.5 13 15 14 0 1 0.05 1.3 15 15 0 3 B 0.5 13 16 14 0 0 0.05 1.3 16 13 0 0

Tricaprylin A 0.1 ml - 15 15 0 0 B 0.1 ml - 16 16 0 1

No Treatment A - - 15 15 0 0 B - - 32** 23 0 3

*Injected SC, once weekly, for 26 weeks in inguinal area. +From beginning of experiments. Median survival ranged from 15 to 21 months in test and control groups. *Laboratory A used ICR/Ha Swiss Millerton female mice. SLaboratory B used CFW Swiss Webster female mice. GINO. in parentheses indicates month when found. **Some mice sacrificed for lung tumor observations to 6 months.

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CLINICAL ASSESSMENT OF SAFETY

Ingestion

In assessing the potential hazards of ingesting Butyl Stearate when the compound was incorporated into food-wrapping film, Smith(“) calculated that a daily dose of 0.2 mg/kg would present little, if any, hazard to humans. Since rats chronically tolerated more than 14,000 times this amount without toxic effects (2500 and 6000 mg/kg daily for 2 years-see Chronic Toxicity section), the minimum safety factor to humans, according to the author, was in excess of 1400. Smith considered this value conservative.

Skin Irritation, Skin Sensitization, Phototoxicity, and Photosensitization

Results from a number of clinical studies indicate that the Stearates and cosmetic products containing the Stearates are essentially nonsensitizing, nonphototoxic, nonphotosensitizing, and at most minimal or mild skin irritants. These clinical tests are individually discussed below, and results have been summarized in Table 9. Butyl, Cetyl, and lsocetyl Stearate were tested for skin irritation and sensitization using a panel of 111 Caucasian women. The concentration of each ingredient was 50 percent in mineral oil. The test materials were applied for 48 hours under semiocclusion to the intact skin of the back. A total of 10 consecutive induction patches were made. Following a 2-week nontreatment period, a 48-hour challenge patch was applied to the back. Butyl Stearate produced, at most, mild skin irritation in 10 subjects and sensitization in 2 subjects. Cetyl Stearate produced, at most, mild skin irritation in 10 subjects and sensitization in 1 subject. The lsocetyl ester caused mild irritation reactions in 10 subjects and no sensitization reactions. Many of the individuals with skin reactions had reactions to more than 1 Stearate. Reactions to the induction patches in any 1 individual were sporadic and did not occur throughout the entire patch series. In no instance were induction or challenge scores greater than 1 or 2 on a scale of 0 (no reaction) to 5. There were no reported areas of abnormal skin pigmentation.“‘) The procedures described by Jordan(gs’ and Jordan and King(99) were used to evaluate the skin irritating and sensitizing properties of both undiluted lsobutyl Stearate and undiluted lsocetyl Stearate. One hundred forty-nine men and women were selected for the study. Each cosmetic ingredient was applied under occlusive 48-hour patches to the skin of the back 3 times weekly for 3 weeks (9 induction applications). Fourteen days following the final induction application, 2 consecutive 48-hour challenge patches of the test material were applied to previously untreated sites of the back. Skin responses were evaluated on a scale of 0 (no reaction) to 4 (bullae or extensive erosions). For lsobutyl Stearate, the number of panelists having a 1+ reaction (macular, faint erythema involving at least 25 percent of the test area) was 2, 1, and 3 to induction application 3, 4, and 8, respectively. One individual had a l+ reaction to the second challenge patch, and a second subject had a 2+ reaction (moderately intense erythema involving at least 25 percent of the test area) to the second challenge patch. No other skin

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TABLE 9. Clinical Studies E Stearate No. of Jest Material Jested Concentration (%) Subjects Method Comments/Results Reference

Skin Irritation/ Butyl, Cetyl, and Iso- Each at 50 111 Ten 48-hour induction Each Stearate was at most 97 Sensitization cetyl Stearate each in patches followed by mildly irritating and es- mineral oil a 2-week nontreat- sentially nonsensitizing ment period, then a to the skin. No abnor- 48-hour challenge mal skin pigmentation patch observed Skin Irritation/ lsobutyl and lsocetyl Each at 100 149 Repeat insult patch lsobutyl Stearate caused 100 Sensitization Stearate test as described by mild skin irritation and )ordonr981 and jor- was nonsensitizing. Iso- dan and King.‘991 cetyl Stearate was both (Nine 48-hour in- nonirritating and non- duction patches, 14- sensitizing day nontreatment period, two 48-hour challenge patches) Skin Irritation Two samples of lsocetyl Each sample at 100 40 Single insult 24.hour 4/40 subjects developed 9, 101 Stearate (20/sample) occlusive patch minimal skin irritation Phototoxicity/ lsobutyl and lsocetyl Each at 50 23 Repeat insult patch Both Stearates were non- 102 Photosensitization Stearate each in min- test procedure with phototoxic and non- eral oil UVA and/or UVB photosensitizing exposure (4400 I*W/ cm*) 8 Skin Irritation Face cream 2.0 Butyl Stearate Unspecified 24 and 48 hour closed The PII of the formulation 103 ; patch test was 0.03 and 0.11 at 24 and 48 hours, re- 5 spectively Skin Irritation/ Unspecified cosmetic IO Butyl Stearate 54 (RIPT) Ten 48-hour occlusive Product was nonirritating, 104, 105 z Sensitization/ product 10 (photo- patches followed by and nonphotosensi- z 0 Photosensitization sensitiza- an 1 l-day nontreat- tizing ;;; ment period, then a 5 48-hour challenge patch. UV treatments were made daily for 10 exposures

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Skin Irritation Personal cleanliness 1 .O Isopropyl Stea- 12 Applied to skin every Formulation was classified 106 product rate day for 21 days by the investigators as under Webril slightly irritating patches Skin Sensitization Two personal cleanli- Each product: 1.0 105 Applied to skin for 24 No skin reactions ob- 106 ness products Isopropyl Stea- hours every other served. Investigators rate day for 3 work concluded that ,, weeks (that is, 10 none of the test applications) materials. possess any potential for skin sensitization under conditions of normal usen Skin Irritation/ Personal cleanliness 1 .O Isopropyl Stea- 40 “To use under normal No adverse reactions ob- 106 Sensitization product rate conditions for 4 served weeks” Skin Irritation Face cream 2.35 Myristyl Stea- 100 Prophetic patch test; No skin reactions 107 rate 50 open and 50 closed patches Skin Irritation Face cream 2.98 Myristyl Stea- 100 Prophetic patch test; No skin reactions 107 rate SO open and 50 closed patches Skin Irritation Underarm cream 4.75 Myristyl Stea- 22 Applied to skin on 3 No skin reactions 107 rate consecutive days Skin Irritation Facial makeup stick 9.8 Myristyl Stea- 100 Prophetic patch test; No skin reactions 107 rate 50 open and 50 closed patches Skin Irritation Facial makeup stick 9.8 Myristyl Stea- 100 Prophetic patch test; No skin reactions 107 rate all patches were closed Skin Irritation/ Suntan lotion and pro- Each product: 7.6 56 Applied to skin every- No skin reactions ob- 108 Sensitization tective face cream Octyl Stearate day under 24.hour served during induction closed patches for or challenge phases. In- total of 10 induction vestigators concluded applications; 24s that the 2 products hour challenge were not capable of in- patch applied after ducing significant irrita- 1 O-l 4-day nontreat- tion or sensitization in ment period humans E

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TABLE 9. (Continued)

Stearate No. of Jest Material Jested Concentration (%) Subjects Method Comments/Results Reference

Phototoxicity Suntan lotion and pro- Each product: 7.6 10 Tape-stripped skin ex- No reactions noted at any 108 tective face cream Octyl Stearate posed to 0.2 g of time during the test. each product for 24 The investigators con- hours under closed cluded that the 2 prod- patch and to UV ucts were not capable light (4400 pW/cm’) of inducing significant for 15 minutes phototoxic reactions in humans Photosensitization Suntan lotion and pro- Each product: 7.6 27 Skin exposed to 0.2 g “Slight reactions” ob- 108 tective face cream Octyl Stearate of each product for served in 4 subjects 24 hours under during the induction closed patches and exposures; 1 of these 4 to UV light (4400 developed “erythema” pW/cml) for 15 min- at challenge. The pro- utes; 24-hour chal- tective face cream eli- lenge patches ap- cited reactions in 3 sub- plied after IO-14- jects during the induc- day nontreatment tion phase; no reactions period, sites irradia- were observed at chal- ted as before lenge. It was the investigator’s opinion that the 2 products were not capable of inducing significant photoallergic reactions in humans

Supplement Panel Book 1 Page 88 Alkyl Esters Supplement Book 1 FINAL REPORT:SAFETY ASSESSMENT OF STEARATES 137 reactions to lsobutyl Stearate were observed. No reactions to lsocetyl Stearate were noted with the exception of 1 individual who developed a 1 + reaction to the initial challenge application. The investigator concluded that both lsobutyl and lsocetyl Stearate were nonsensitizing.““’ A single insult, 24hour occlusive patch test was performed with 2 samples (A and B) of undiluted lsocetyl Stearate on 40 subjects (20 subjects/sample). Of the 20 subjects tested with sample A, 2 had barely perceptible to mild skin erythema, and 18 had no skin irritation. Of the 20 subjects exposed to sample B, 2 had “barely perceptible” to mild skin erythema, whereas 18 had no skin irritation. The PII to each sample was 0.08 (maximum, 4.0), indicating minimal skin irritation (9.101) Twenty-three volunteers (17 women and 6 men) participated in a study designed to evaluate the phototoxicity and photosensitization of both lsobutyl and lsocetyl Stearate. Each cosmetic ingredient was prepared as a 50 percent concentration in mineral oil and subsequently tested in an identical manner. A 24hour occlusive patch containing 0.1 ml of the test material was applied to the same site on the back of each subject on Monday, Wednesday, and Friday for 3 consecutive weeks (9 induction patches). Challenge patches were applied 2 weeks later to both the original site and to a previously untreated, adjacent site. Identical patches were also applied to the opposite side of the back to serve as nonirradiated controls. Of the 23 subjects tested, 9 received both UVA (320 to 400 nm) and UVB (280 to 320 nm) exposure, whereas 14 subjects received UVA only. The UVA exposure (50 percent transmission at 345 nm) was delivered by 4 BL fluores- cent tubes at 10 cm from the skin at a dose of approximately 4400 pW/cm*. Sites were irradiated for 5 minutes after patch removal on each Tuesday and Saturday during the induction phase. Additionally, a 150 W Xenon Arc Solar Simulator was used to deliver twice the individual minimal erythemal dose (MED) of UVB to those designated subjects after each UVA exposure. At challenge, test sites were exposed only to UVA irradiation. Skin responses were scored on a scale of 0 (no reaction) to 4 (erythema and papules) just prior to each new patch application and 72 hours after the last induction patch. Challenge sites were scored at 24, 48, 72, and 96 hours. For both lsobutyl and lsocetyl Stearate, the combination of UVA and UVB exposure produced erythematous reactions (score, 1) in all 9 subjects during the induction phase. According to the investigator, this was an expected result as the subjects were exposed to 2 times their MED. For lsobutyl Stearate, 2 of these 9 individuals (No. 4 and No. 20) had 3 reactions (erythema and edema) as a result of the first induction exposure. However, it was determined that the 2 subjects had received an excessive dose of UVB. The time of UVB exposure was subsequently reduced for these 2 individuals. Of the 2 subjects having a 3 reaction to the initial induction exposure, 1 had a second 3 reaction on the fourth induction exposure; all other scores during induction for this individual were no higher than 1. The second subject with an initial 3 induction score to lsobutyl Stearate had scores no higher than 1 throughout the remainder of the induction period. For lsocetyl Stearate, 3 subjects (No. 4, No. 13, No. 20) had 3 reactions to the first induction exposure due to an excessive dose of UVB irradiation. When the UBV exposure time was reduced for these 3 individuals, no induction reaction above 1 was noted. No other skin reactions were observed on irradiated or nonirradiated sites to either ingredient during induction or chal-

Supplement Panel Book 1 Page 89 Alkyl Esters Supplement Book 1 138 COSMETIC INGREDIENT REVIEW lenge phases. It was concluded that both lsobutyl and lsocetyl Stearate, each at 50 percent in mineral oil, were neither phototoxic nor photosensitizing agents in humans. (lo’) A face cream containing 2.0 percent Butyl Stearate was tested for skin irritation on an unspecified number of subjects. The formulation was applied to the skin full strength under closed patches for either 24 or 48 hours. PIJ was 0.03 in subjects exposed for 24 hours and 0.11 in those exposed for 48 hours. No other details were reported.(lo3) An unspecified type of cosmetic product containing 10 percent Butyl Stea- rate was found nonirritating and nonsensitizing to the skin in a repeat insult patch test (RIPT) with a panel of 54 women. (‘04) The same product was also nonphotosensitizing in a test involving UV exposure. (lo’) The induction phase of the RIPT procedure called for application of 10 consecutive 48-hour occlusive patches containing the product to the back of each of 10 women. Following an 1 l-day nontreatment period, a 48-hour challenge patch was applied to a previously un- exposed site on the back. Application sites were graded following patch removal. In the photosensitization test, application of the product containing 10 percent Butyl Stearate’to the same areas with UV exposures was made daily for ten treatments.” Additional details of the photosensitization test were lacking. A personal cleanliness product containing 1 percent Isopropyl Stearate was tested for skin irritation on 12 subjects (1 male, 11 females). The formulation was applied under an occlusive patch once a day for 21 days to the skin of the back. The test material was reapplied to the same site throughout the study. Prior to application, the formulation was sprayed onto the patch and allowed to evaporate 10 minutes. Twenty-three hours after application, the patch was removed. The panelist was then instructed to bathe or shower immediately following removal of the patch. Reactions to the product were scored 1 hour after patch removal. Following the twenty-first treatment, a cumulative skin irritation score was calculated, and the test material was classified as one of the following: essentially nonirritating, slightly irritating, moderately irritating, or highly irritating. Under conditions of this test, the “personal cleanliness” product containing 1 percent Isopropyl Stearate was classified as slightly irritating.(‘06’ The skin-sensitizing potential of 2 personal cleanliness products each containing 1 percent Isopropyl Stearate was tested using 105 subjects. For the induction phase of the study, each product was applied to the skin of the back for 24 hours every other day for “three work weeks” (10 applications). The only exception to this procedure was that the third application of each week followed the second by only 10 hours; this was in contrast to the 24hour interval between first and second applications. The test material was applied to the same site throughout the induction period. Fourteen days following the tenth induction patch, a 48-hour challenge patch was applied to the original site. A second 48-hour challenge patch was applied 8 days after the first. During the course of the study, no skin reactions were observed. The investigators concluded that “. . . none of the test materials. . . possess any potential for skin sensitization under conditions of normal use.“(lo6) Forty subjects were given a personal cleanliness product to use “under normal conditions” for 4 weeks. The formulation contained 1 percent Isopropyl Stea- rate. No adverse reactions were observed.(106)

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Cosmetic products containing various concentrations of Myristyl Stearate were tested in 5 separate studies for skin irritation. A face cream containing 2.35 percent Myristyl Stearate and a second face cream containing 2.98 percent Myris- tyl Stearate were each applied to the skin of 100 subjects (200 total) in 2 prophetic patch tests. Fifty open and 50 closed patches were used in each study. No skin reactions were noted with either product. In another study, an underarm cream containing 4.75 percent Myristyl Stearate was applied to the skin of 22 subjects for 3 consecutive days. No skin irritation was observed. Two facial makeup sticks each formulated with 9.8 percent Myristyl Stearate were evaluated using groups of 100 subjects. The prophetic patch procedure called for 50 open and 50 closed patches in 1 study (100 subjects) and “all closed patches” in the other (100 subjects). No skin reactions were observed.(107) A suntan lotion and a protective face cream each formulated with 7.6 percent Octyl Stearate were tested for skin irritation and/or sensitization. Approxi- mately 0.2 g of each product was applied for 24 hours under closed patches to the inner aspect of the arm or skin of the back of 56 subjects. Patches were applied to the same site on Mondays, Wednesdays, and Fridays until a total of 10 induction applications had been made. Ten to fourteen days after the final induction patch, challenge patches were applied for 24 hours to both the original contact site and to a previously untreated adjacent site. Test sites were scored 24 and 48 hours after application. No skin reactions were observed during the induction or challenge phase to either of the product formulations. The investigator concluded that the 2 products did not produce significant irritation or sensitization in humans.(108) To determine phototoxicity to a suntan lotion and protective face cream, 10 subjects were selected for study. Test sites on the inner aspect of the forearm were tape-stripped 6 to 10 times to remove several layers of cornified epithelium. Each product (0.2 g) was then applied under closed patches to control and test sites for 24 hours. At the end of this period, test sites were subjected to UV irradiation at a dose of 4400 PWlcm’ for 15 minutes. The UV light source, held 10 to 12 cm from the skin, consisted of 4 GE F40 BL black light lamps. The wavelength of the light source was in the UVA range, with a peak at 360 nm. No reactions were noted to either product containing 7.6 percent Octyl Stearate. The investigator concluded that the 2 products did not produce significant phototoxic reactions in humans.(108) Twenty-seven subjects were tested with a suntan lotion and protective face cream for photosensitization. Approximately 0.2 g of each product was applied to test sites under closed patches for 24 hours. Following removal of the patch, test sites were given nonerythrogenic UV irradiation at a dose of 4400 pW/cm* for 15 minutes. The UV light source consisted of a GE F40 BL black light with a peak wavelength of 360 nm. The control sites were covered with black tape to prevent UV exposure. Immediately following irradiation, all sites were evaluated. The patch sites were then covered with Webril or Dermicel to prevent inadvertent exposure to sunlight. This procedure was repeated each Monday, Wednesday, and Friday until “10 applications/irradiation” had been made. Following a 10 to 14day nontreatment period, challenge patches were applied to the original contact site and to a previously untreated adjacent site. Twenty-four hours after application, the patches were removed, the sites examined for dermal response,

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140 COSMETIC INGREDIENT REVIEW

and the test areas irradiated again. Additional scorings were made 48 and 72 hours after application. The suntan lotion containing 7.6 percent Octyl Stearate produced “slight reactions” in 4 subjects. One subject developed “slight delayed reactions” to the sixth induction exposure at both control and irradiated test sites. However, no further reactions were observed at either site. A second and third subject had “slight reactions” to the ninth and tenth induction exposures, respectively. Neither of these 2 people reacted to the challenge patches. A fourth subject developed “erythema” as a result of the eighth induction exposure; this reaction dissipated after the site had been irradiated. This same subject also developed “erythema”at each of the 2 challenge sites. According to the investigators, the skin reactions of the fourth subject were “only slight and probably not significant.” Three subjects reacted to the protective face cream containing 7.6 percent Octyl Stearate. Two subjects had erythema to the sixth and eighth induction exposures; 1 of these subjects “reacted” after the seventh induction patch at the nonirradiated control site. A third subject”reacted slightly” to the ninth induction exposure. No reactions were observed in any subjects at challenge. It was the investigator’s opinion that the 2 product formulations containing 7.6 percent Octyl Stearate were not capable of inducing significant photoallergic reactions in humans. (loa)

SUMMARY

The 7 Stearates described in this report are either oily liquids or waxy solids. They are prepared by esterification of stearic acid with the appropriate alcohol. Because of the technical grade of stearic acid and the alcohols used as starting materials, the commercially available stearates are often mixtures of various esters. These Stearates can be expected to undergo reactions typical of esters, such as conversion into stearic acid and the corresponding alcohol by chemical or enzymatic hydrolysis, conversion into amides by ammonolysis, and conversion into different esters by alcoholysis or transesterification. Butyl, Isobutyl, and lsocetyl Stearate have a wide variety of noncosmetic applications. Some of these applications include use as a lubricant, solvent fixative, and/or emollient in pharmaceuticals, as a solvent for dyes, as a spreading and/or softening agent in textiles, plastics, and rubber, as a plasticizer, and as a waterproofing agent. Federal regulations permit the use of Butyl Stearate as a direct and indirect food additive and as a synthetic flavoring agent. The Stearates are primarily used in cosmetics as skin emollients. Depending on the specific type of cosmetic product, these esters may also function as color- suspending agents, water-repelling plasticizers, spreading or “carrying” agents, stiffening agents, gelatin enhancers, lubricants, fixatives, solvents, or viscosity builders. When applied to the skin or lips, the low viscosity and oily nature of these ingredients provide a nongreasy, hydrophobic film. Cosmetic firms participating in the voluntary cosmetic registration program reported to the FDA in 1981 that Butyl, Cetyl, Isocetyl, Isopropyl, Myristyl, and Octyl Stearates were used in a total of 116, 4, 58, 16, 1, and 10 cosmetic products, respectively. Stearate concentrations in these products generally ranged from 1 to 25 percent, although there were a few reported instances of higher and lower concentrations. The most frequent uses of the Stearates were in eye

Supplement Panel Book 1 Page 92 Alkyl Esters Supplement Book 1 FINAL REPORT:SAFETY ASSESSMENTOF STEARATES 141 makeup preparations, skin makeup preparations, lipstick, and skin care preparations. Cosmetics containing these esters are intentionally applied to or come in contact with eyes, skin, hair (and scalp), nails, and vaginal mucosa. Small amounts of these esters could be ingested from lipstick. Aliphatic esters are hydrolyzed to the corresponding alcohol and acid and further metabolized. In the case of the Stearate ingredients, stearic acid would be metabolized in the same manner as other fatty acids. The toxicology of the Stearates was assessed in a number of animal studies. Butyl Stearate (50 percent in mineral oil) was weakly comedogenic when applied to rabbit skin 5 days a week for 2 weeks. The subcutaneous and intraperitoneal LDsos in rats of undiluted Butyl Stearate were both >32 mglkg. Acute oral LDso values varied according to ingredient and species tested. However, Butyl, Iso- cetyl, Isopropyl, Myristyl, and Octyl Stearates generally had low acute oral toxicity in rats and mice. Undiluted Butyl and Isopropyl Stearate was nonirritating to the rabbit eye, whereas undiluted Isocetyl, Myristyl, and Octyl Stearates caused slight transient ocular irritation in rabbits. Undiluted Butyl, Isobutyl, Isocetyl, Iso- propyl, Myristyl, and Octyl Stearates produced at most minimal or moderate skin irritation in rabbits. Application of undiluted Octyl Stearate to the skin of rabbits daily for 60 days caused irritation (vesicles and slight epidermal exfoliation) and was generally “poorly tolerated.” Microscopic changes in the treated skin included epidermal acanthosis and “congestive” dermatitis. Application of 10 percent Octyl Stearate in aqueous solution to rabbit skin daily for 60 days caused irritation (vesicles) but was “relatively well tolerated.” No significant pathological reactions were observed upon microscopic examination of the treated skin. No skin sensitization was observed to either Butyl (0.1 percent in physiological saline) or lsocetyl Stearate (0.1 percent in physiological saline) when given intracutaneously to guinea pigs by means of 10 induction injections and 1 challenge injection. Fertility, litter size, and survival of offspring were normal in rats fed diets containing 6.25 percent Butyl Stearate for 10 weeks. However, growth was reduced in offspring during the preweaning and postweaning period. No gross lesions were noted among the offspring killed at the end of the 21-day postweaning period. Rats fed dietary concentrations of 1.25 or 6.25 percent Butyl Stearate (corresponding to 2500 and 6000 mg/kg, respectively) for 2 years had no significant differences from control animals with respect to growth, survival, blood counts, or other hematologic parameters. In clinical studies, Butyl, Cetyl, Isobutyl, and lsocetyl Stearates (undiluted or 50 percent in mineral oil) and cosmetic products containing Butyl, Isopropyl, Myristyl, and Octyl Stearates (1 .O, 2.0, 2.35, 2.98, 4.75, 7.6, and/or 9.8 percent) were at most minimally to mildly irritating to the human skin, essentially nonsensitizing, nonphototoxic, and nonphotosensitizing. It has been estimated by 1 investigator that daily ingestion of 0.2 mg/kg Butyl Stearate would present little, if any, hazard to humans. This estimation was based on the results of chronic feeding studies in rats.

DISCUSSION

Currently available tests are inadequate in predicting the ability of a cosmetic ingredient to cause comedones in humans. However, comedogenicity is a poten-

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142 COSMETIC INGREDIENT REVIEW tial health effect that should be considered when the Stearate ingredients are used in cosmetic formulations.

CONCLUSION

On the basis of the information presented in this report, the CIR Expert Panel concludes that Butyl, Cetyl, Isobutyl, Isocetyl, Isopropyl, Myristyl, and Octyl Stearate are safe as cosmetic ingredients in the present practices of use.

ACKNOWLEDGMENT

Jonathon T. Busch, Senior Scientific Analyst, prepared the literature review and technical analysis used to develop this report.

REFERENCES

1. COSMETIC, TOILETRY AND FRAGRANCE ASSOCIATION. (CTFA). (Aug. 24, 1979). Submission of unpublished data by CTFA. Cosmetic ingredient descriptions: lsocetyl Stearate, Isopropyl Stearate, Octyl Stearate, and Stearyl Stearate.* 2. CTFA. (Aug. 30, 1979). Submission of unpublished data by CTFA. Cosmetic ingredient chemical descriptions: Butyl Stearate and lsobutyl Stearate.* 3. ESTRIN, N.F., CROSLEY, P.A., and HAYNES, C.R. (1982). CTFA Cosmetic Ingredient Dictionary, 3rd ed. Washington, DC: Cosmetic Toiletry and Fragrance Association. 4. RESEARCH INSTITUTE FOR FRAGRANCE MATERIALS (RIFM). (August 1979). Food and Cosmetics Toxi- cology. Fragrance Raw Materials Monographs. Stearic Acid. 17(4), 383-8. 5. FEDERATION OF AMERICAN SOCIETIES FOR EXPERIMENTAL BIOLOGY (FASEB). (1975). Select Com- mittee on GRAS Substances. Evaluation of the health aspects of tallow, hydrogenated tallow, stearic acid, and calcium stearate as food ingredients. FDA Contract 233-75-2004. 6. INFORMATICS, INC. (July 1973). Scientific literature reviews on generally recognized as safe (GRAS) food ingredients, tallow and stearic acid. Prepared for the Food and Drug Administration under Contract Grant No. FDA-72-104. NTIS Publication No. PB 223859. 79 pp. 7. ESTRIN, N.F. fed.). (1977). CJFA Cosmetic ingredient Dictionary, 2nd ed. Washington, DC: Cosmetic, Toiletry and Fragrance Association. 8. NATIONAL LIBRARY OF MEDICINE (NLM). (1983). Dept. of Health and Human Services. Bethesda, MD. Computerized Data Bank (Medlars). Files: Chemline, Toxicology Data Bank, Registry of Toxic Effects of Chemical Substances. 9. CTFA. (June 11, 1980). Submission of unpublished data by CTFA. Summary of unpublished safety data on the lsocetyl Stearate group.* 10. HAWLEY, G.G. fed.). (1971). The Condensed Chemical Dictionary, 8th ed. New York: Van Nostrand Reinhold. 11. WINDHOLZ, M. (ed.). (1976). The Merck Index, 9th ed. Rahway, NJ: Merck. 12. FURIA, T.E., and BELLANCA, N. (eds.). (1971). Fernaroli’s Handbook ofFlavor Ingredients, 2nd ed. Cleve- land, OH: CRC Press, Vol. 2. 13. ESTRIN, N.F. (ed.). (May 1971). CTFA Standards. Cosmetic ingredient specifications: Butyl Stearate, Cetyl Alcohol, Myristyl Alcohol, Stearyl Alcohol. Washington, DC: Cosmetic Toiletry and Fragrance Associa- tion.”

*Available upon request: Administrator, Cosmetic Ingredient Review, Suite 810, 1110 Vermont Avenue, N.W., Washington, DC 20005.

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FINAL REPORT:SAFETY ASSESSMENT OF STEARATES 143

14. ESTRIN, N.F., HAYNES, C.R., and WHELAN, J.M. (1982). CTFA Compendium of Cosmetic ingredient Compositions Specifications/Spectra. Washington, DC: Cosmetic, Toiletry and Fragrance Association. 15. ESTRIN, N.F. (ed.). (Oct. 15, 1974). CJFA Standards. Cosmetic ingredient specifications: lsocetyl Alcohol. Washington, DC: Cosmetic, Toiletry and Fragrance Association. 16. CTFA. (Feb. 26, 1979). Submission of unpublished data by CTFA. Cosmetic ingredient chemical description: Stearic Acid.* 17. ESTRIN, N.F., HAYNES, C.R., and WHELAN, J.M. (1982). CJFA Compendium of Cosmetic lngredietit Composition. Cosmetic ingredient descriptions, Washington, DC: Cosmetic, Toiletry and Fragrance Asso- ciation. 18. GREENBERG, L.A., LESTER, D., and HAGGARD, H. (1954). Handbook of Cosmetic Materials. New York: Interscience Publishers. 19. PATTY, F.A. (ed.). (1963). fndustrial Hygiene and Toxicology, 2nd ed. New York: Interscience Publishers, Vol. 2. 20. YAKUJI NIPPO, LTD. (1979). Japanese Standards of Cosmetic Ingredients. The Japan Cosmetic Industry Association, 1-l 1 Kanda lzumicho, Chiyoda-ku, Tokyo, 101, Japan. 21. LOWER, E.S. (1982). Butyl Stearate-A review. Manuf. Chem. Aerosol News 55, 57, 59-60. 22. ESTRIN, N.F. (ed.). (1971). CJFA Standards. Spectra: Butyl Stearate. Washington, DC: Cosmetic, Toiletry and Fragrance Association. 23. ESTRIN, N.F. (ed.). (1974). CJFA Standards. Cosmetic ingredient descriptions: lsocetyl Stearate. Washing- ton, DC: Cosmetic, Toiletry and Fragrance Association. 24. MORRISON, R.T., and BOYD, R.N. (1973). Organic Chemistry, 3rd ed. Boston, MA: Allyn and Bacon. 25. SAX, N.I. (ed.). (1968). Dangerous Properties of Industrial Materials, 3rd ed. New York: Reinhold Book Corp. 26. CROPPER, F.R., and HEYWOOD, A. (1953). Analytical separation of the methyl esters of the C12C22 fatty acids vapor-phase chromatography. Nature 172, 1101-2. 27. LINK, W.E. (ed.). (1971-2). Official and Tentative Methods of the American 011 Chemists’ Society, 3rd ed. Champaign, IL: American Oil Chemists’ Society. 28. GROSS, F.D. (1966). Gas-liquid chromatography of some fatty acid esters and alcohols in lipsticks. J. As- soc. Off. Anal. Chem. 49(6), 1195-200. 29. HORNSTEIN, I,, ELLIOTT, L.E., and CROWE, P.F. (1959). Gas chromatographic separation of long-chain fatty acid methyl esters on poly(vinylacetate). Nature 184 [Suppl. No. 221, 1710-l 1. 30. KRUPCIK, J., TESARIK, K., LISKA, O., NEMEC, J., and DUCHESNE, Y.P. (1972). Effect of the support on the gas-chromatographic separation of fatty acid methyl esters. Chromatographia 5(1 l), 252-7. 31. MURATA, T. (1978). Analysis of fatty acid methyl esters by a gas-liquid chromatography-chemical ionization mass spectrometry computer system. J. Lipid Res. 19(2), L166-71. 32. ORR, C.H., and CALLEN, J.E. (1958). Separation of polyunsaturated fatty acid methyl esters by gas chromatography. J. Am. Chem. Sot. 80, 249. 33. WATANABE, S., NAKASATO, S., KUWAYAMA, H., SASAMOTO, Y., SHIRAISHI, S., SEINO, H., NAGAI, T., NEGISHI, M., and HAYANO, 5. (1973). Determination of fatty acid composition using a gas chromatography with a flame ionization detector. Yukagaku 22(2), 95-101. 34. RAMSEY, J.D., LEE, T.D., OSSELTON, M.D., and MOFFAT, A.C. (1980). Gas-liquid chromatographic retention indices of 296. Nondrug toxicological analyses. J. Chromatogr. 184(2), 185-206. 35. LIE KEN JIE, M.S.F., and LAM, C.H. (1977). Fatty acids. XV. Thin-layer chromatographic behavior of acetylenic fatty esters on silicic acid. J. Chromatogr. 136(l), 178-81. 36. MADELMONT, C., and PERRON, R. (1968). Chromatography on a layer of silica gel activated carbon application to analytical and preparative separation of saturated fatty-acids and their methyl esters. Qual. Plant Mat. Veg. 16(1-4), 126-30. 37. OSMAN, F.. SUBBARAM, M.R., and ACHAYA, K.T. (1967). Quantitative microthinlayer chromatography of fatty materials. J. Chromatogr. 26(l), 286-9. 38. RAKOFF, H., and EMKEN, E.A. (1978). Silver resin chromatographic separation of methyl cis- and trans- mono- and dihydroxy fatty esters, J. Am. Oil Chem. Sot. 55(7), 564-6. 39. SCHOLFIELD, C.R., and MOUNTS, T.L. (1977). New developments in silver resin chromatography ofcis and trans fatty methyl esters, J. Am. Oil Chem. Sot. 54(8), 319-21. 40. SCHOLFIELD, C.R. (1979). Silver nitrate-high performance liquid chromatographyoffatty methyl esters. J. Am. Oil Chem. Sot. 56(4), 510-l. 41. CASTELLANI, T., KJELGAARD-NIELSEN, P., and WOLFF-JENSEN, J. (1975). Relationship between densito- metric peak area and concentration of lipid in the quantitative analysis of a lipid mixture separated by thin-layer chromatography. J. Chromatogr. 104(l), 123-34.

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144 COSMETIC INGREDIENT REVIEW

42. LUTZ, D.A., EDDY, C.R., and HUNTER, J.J. (1967). X-ray diffraction study of some normal alkyl esters of long-chain acids. Lipids 2(3), 204-7. (317s) 43. CODE OF FEDERAL REGULATIONS. (CFR). (Revised as of April 1, 1982). Title 21 part 175.105. Adhe- sives. 44. CFR. (Revised as of April 1, 1982). Title 21 Part 175.300. Resinous and polymeric coatings. 45. CFR. (Revised as of April 1, 1982). Title 21 Part 175.320. Resinous and polymeric coatings for polyolefin films. 46. CFR. (Revised as of April 1, 1982). Title 21 Part 176.200. Defoaming agents used in coatings. 47. CFR. (Revised as of April 1, 1982). Title 21 Part 177.2600. Rubber articles intended for repeated use. 48. CFR. (Revised as of April 1, 1982). Title 21 Part 178.3910. Surface lubricants used in the manufacture of metallic articles. 49. CFR. (Revised as of April 1, 1982). Title 21 Part 181.27. Plasticizers. 50. CFR. (Revised as of April 1, 1982). Title 21 Part 173.340. Defoaming agents. 51. CFR. (Revised as of April 1, 1982). Title 21 Part 172.515. Synthetic flavoring substances and adjuvants. 52. BALSAM, M.S., and SAGARIN, E. (eds.). (1972). Cosmetics: Science and Technology, 2nd ed., Vol. 1. Stri- anse, S.J.: Hand Creams and Lotions. Lauffer, P.G.I.: Lipsticks. New York: Wiley-Interscience. 53. FOOD AND DRUG ADMINISTRATION (FDA). (Dec. 22, 1981). Cosmetic product formulation data. In- gredient used in each product category. Computer printout. Washington, DC. 54. FDA. (Jan. 5, 1982). (a) Frequency of trade name ingredients in the cosmetic product file and (b) frequency of common, usual, or chemical names in the cosmetic product file. Computer printout, Washing- ton, DC. 55. CFR. (Revised as of April 1, 1982). Title 21 Part 720.4. Voluntary filing of cosmetic product ingredient and cosmetic raw material composition statement. Information requested about cosmetic products. 56. SAVARY, P., and CONSTANTIN, M.J. (1970). Intestinal hydrolysis and lymphatic absorption in isopropyl esters of long-chain fatty acids in the rat. Biochim. Biophys. Acta 218(2), 195-200. 57. KLICMAN, A.M., and MILLS, O.H., JR. (1972). Acne cosmetica. Arch. Dermatol. 106(6), 843-50. 58. SMITH, C.C. (1953). Toxicity of butyl stearate, dibutyl sebicate, dibutyl phthalate and methoxyethyl oleate. Arch. Ind. Hyg. Occup. Med. 7(4), 310-8. 59. AVON PRODUCTS, INC. (Dec. 22, 1976). Submission of unpublished data by CTFA. Biological evaluation summary report: Skin and eye irritancy and acute toxicity. Butyl Stearate.’ 60. INOLEX CORP. (April 29, 1975). Submission of unpublished data by CTFA. Acute oral toxicity test. Butyl Stearate.* 61. WICKHEN PRODUCTS, INC. (Oct. 6, 1972). Submission of unpublished data by CTFA. Acute oral toxicity of Wickenol 122. Butyl Stearate.* 62. EMERY INDUSTRIES, INC. (Dec. 1, 1975). Submission of unpublished data by CTFA. Eye irritation. Emerest 2325. Butyl Stearate. Leberco Laboratories.* 63. ARMAK CO. (June 9, 1978). Submission of unpublished data by CTFA. Kessco lsocetyl Stearate, 2732002.j 64. VAN DYK AND CO., INC. (Feb. 21, 1979). Submission of unpublished data by CTFA. Acute oral toxicity, primary dermal irritation, and ocular irritation. Ceraphyl494. lsocetyl Stearate. Consumer Product Testing Co., Inc.* 65. INOLEX CORP. (April 4, 1979). Submission of unpublished data by CTFA. Acute oral toxicity. Lexol HDS. lsocetyl Stearate.* 66. WICKHEN PRODUCTS, INC. (Sept. 27, 1972). Submission of unpublished data by CTFA. Acute oral toxicity of Wickenol 127. Isopropyl Stearate.* 67. VAN DYK AND CO., INC. (Jan. 26, 1971). Submission of unpublished data by CTFA. Acute oral toxicity. Myristyl Stearate. Leberco Laboratories.* 68. WICKHEN PRODUCTS, INC. (Oct. 20, 1972). Submission of unpublished data by CTFA. Acute oral toxicity of Wickenol 156. Octyl Stearate. Kolmar Research Center.* 69. EMERY INDUSTRIES, INC. (Aug. 15, 1974). Submission of unpublished data by CTFA. Draize eye irritation, primary skin irritation, and acute oral LDso toxicity studies. Emerest 2325, Butyl Stearate. Bio-Toxi- cology Laboratories, Inc.* 70. INOLEX CORP. (April 30, 1979). Submission of unpublished data by CTFA. Draize eye irritation test. Lexol HDS. lsocetyl Stearate.* 71. DRAIZE, J.H., WOODARD, C., and CALVERY, H.O. (1944). Methods for the study of irritation and toxicity of substances applied topically to the skin and mucous membranes. J. Pharmacol. Exp. Ther. 82, 377. 72. MB RESEARCH LABORATORIES, INC. (Jan. 25, 1977). Submission of unpublished data by CTFA. Report on rabbit eye irritation. Project No. MB 77-2290. One hundred percent Isopropyl Stearate.”

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FINAL REPORT:SAFETY ASSESSMENT OF STEARATES 145

73. VAN DYK AND CO., INC. (Jan. 18, 1971). Submission of unpublished data by CTFA. Eye irritation. Myris- tyl Stearate. Leberco Laboratories.* 74. GUILLOT, J.P., MARTINI, M.C., and CIAUFFRET, J.Y. (1977). Safety evaluation of cosmetic raw materials. J. Sot. Cosmet. Chem. 28, 377-93. 75. JOURNAL OFFICIEL DE LA REPUBLIQUE FRANCAISE. (April 21, 1971). Edition Lois et Decrets. Annexe 1: Methode Officielle pour la Determination de L’indice D’irritation Primaire. Annexe 2: Methode Offi- cielle pour la Determination de L’irritation Occulaire. Annexe 3: Methode Officielle pour L’appreciation de L’aggresivite superficielle Cutanee par Applications Interatives, pp. 3862-4. 76. ARMAK CO. (no date). Submission of unpublished data by CTFA. Kessco Butyl Stearate Cosmetic Lot No. 70272. Primary skin irritation.* 77. ARMAK CO. (no date). Submission of unpublished data by CTFA. Kessco Butyl Stearate Cosmetic Lot No. 71352. Primary skin irritation.* 78. AVON PRODUCTS, INC. (Jan. 3, 1980). Submission of unpublished data by CTFA. Toxicology summary report: Animal tests. Butyl Stearate. Primary skin irritation.* 79. CFR. (Revised as of October 1, 1981). Title 49 Part 173.240. Corrosive material; definition. 80. DRAIZE, J.H. (1959). Dermal Toxicity. Appraisal of the Safety of Chemicals in Foods, Drugs and Cos- metics. Div. of Pharmacology, FDA, Dept. of HEW. The Assoc. of Food and Drug Officials of the U.S. Bus- iness Office, Bureau of Food and Drugs, Austin, Texas, pp. 46-59. 81. CFR. (Revised as ofJanuary 1, 1982). Title 16 Part 1500.41. Method of testing primary irritant substances. 82. INOLEX CORP. (April 25, 1975). Submission of unpublished data by CTFA. Primary skin irritation. Butyl Stearate.* 83. WICKHEN PRODUCTS, INC. (Nov., 1972). Submission of unpublished data by CTFA. Primary skin irritation. Butyl and Isopropyl Stearate.* 84. ARMAK CO. (no date). Submission of unpublished data by CTFA. lsobutyl Stearate Lot No. 71222. Pri- mary skin irritation.* 85. AVON PRODUCTS, INC. (April 4, 1974). Submission of unpublished data by CTFA. Biological evaluation summary report: Skin and eye irritancy. Hexadecyl Stearate.* 86. INOLEX CORP. (April 2, 1979). Submission of unpublished data by CTFA. Primary skin irritation. Lexol HDS. lsocetyl Stearate.* 87. CTFA. (November, 1972). Submission of unpublished data by CTFA. Primary irritation ofthe skin (rabbits) and acute oral toxicity (rats). Isopropyl Stearate (100 percent), Lot 22.’ 88. VAN DYK AND CO., INC. (Jan. 14, 1971). Submission of unpublished data by CTFA. Dermal irritation. Myristyl Stearate. Leberco Laboratories.* 89. WICKHEN PRODUCTS, INC. (Dec. 19, 1979). Submission of unpublished data by CTFA. Primary dermal irritation. 2-Ethylhexyl Stearate (Octyl Stearate). MB Research Laboratories, Inc.* 90. INOLEX CORP. (June 5, 1975). Submission of unpublished data by CTFA. Guinea pig sensitivity test. Butyl Stearate. * 91. INOLEX CORP. (May 8, 1979). Submission of unpublished data by CTFA. Guinea pig sensitivity test. Lexol HDS. lsocetyl Stearate.* 92. SWERN, D., WIDER, R., McDONOUCH, M., MERANZE, D.R., and SHIMKIN, M.B. (1970). Investigation of fatty acids and derivatives of carcinogenic activity. Cancer Res. 30(4), 1037-46. 93. VAN DUUREN, B.L., KATZ, C., SHIMKIN, M.B., SWERN, D., and WEIDER, R. (1972). Replication of low- level carcinogenic activity bioassays. Cancer Res. 32(4), 880-l. 94. DEICHMANN, W.B., RADOMSKI, J.L., MACDONALD, W.E., KASCHT, R.L., and ERDMANN, R.L. (1958). The chronic toxicity of octadecylamine. Arch. Indust. Health 18, 483-7. 95. BOYLAND, E., BUSBY, E.R., DUKES, C.E., GROVER, P.L., and MANSON, D. (1964). Further experiments on implantation of materials into the urinary bladder of mice. Br. J. Cancer 18(j), 575-81. 96. INTERNATIONAL AGENCY FOR RESEARCH ON CANCER (IARC). (August 1977). IARC Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to Man, World Health Organization. 15:21. 97. CTFA. (Sept. 30, 1982). Submission of unpublished data by CTFA. Repeat-insult sensitization study. Pro- ject LCR-000. Protocol 38-Z Butyl. Cetyl, lsocetyl and Stearyl Stearate (50 percent concentration).* 98. JORDAN, W.P. (1980). 24., 48-, and 48/48 hour patch tests. Contact Dermatitis 6, 151-2. 99. JORDAN, W.P., and KING, S.E. (1977). Delayed hypersensitivity in females. The development of allergic contact dermatitis in females during the comparison of two predictive patch tests. Contact Dermatitis 3, 19-23. 100. HILL TOP RESEARCH, INC. (Sept. 7, 1982). Submission of unpublished data by CTFA. Jordon-King Modi- fication of the Draize-Shelanski Procedure. Modified Draize repeated insult patch test. lsobutyl and ISO- cetyl Stearate.’

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101. AVON PRODUCTS, INC. (March 6, 1974). Submission of unpublished data by CTFA. Clinical evaluation report: Human patch test. Hexadecyl Stearate.* 102. FOOD AND DRUG HUMAN CLINICAL LABS, INC. (June 28, 1983). Submission of unpublished data by CTFA. Final report: Human photosensitization of samples isobutyl stearate and isocetyl stearate? 103. CTFA. (no date). Submission of unpublished data by CTFA. Data on cosmetic products containing Butyl Stearate. Face Cream Code No. 0135.* 104. TESTKIT LABORATORIES, INC. (Feb. 24, 1981). Submission of unpublished data by CTFA. Repeated insult patch test. Study No. 81.21 Product No. 452-27 containing 10 percent Butyl Stearate.* 105. WELLS LABORATORIES, INC. (Jan. 12, 1981). Submission of unpublished data by CTFA. Report on Pho- tosensitivity Test. Product No. 452-27 containing 10 percent Butyl Stearate.* 106. CTFA. (no date). Submission of unpublished data by CTFA. Summary of human skin irritation and sensitization data on a personal cleanliness product containing Isopropyl Stearate.* 107. CTFA. (no date). Submission of unpublished data by CTFA. Data on cosmetic products containing Myris- tyl Stearate. Underarm cream; facial makeup stick; face cream.* 108. CTFA. (NOV. 14, 1978). Submission of unpublished data by CTFA. Data on cosmetic products containing Octyl Stearate. Clinical evaluation of two products: Suntan lotion and protective face cream. Food and Drug Research Laboratories, Inc.*

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JOURNAL OF THE AMERICAN COLLEGE OF TOXICOLOGY Volume 4, Number 3,198s Mary Ann Liebert, Inc., Publishers

Final Report on the Safety Assessment of lsostearyl Neopentanoate

lsostearyl Neopentanoate, the ester of lsostearyl Alcohol and Neopentanoic Acid, is used in cosmetic products as an emollient at concentrations up to 50 percent. The undiluted ingredient at doses up to 4 ml/kg was shown to be relatively non-toxic in short- and long-term feeding studies. Test data from animal and clinical studies indicate the undiluted ingredient is neither an irritant nor a sensitizer. A cosmetic formulation containing 16 percent lsostearyl Neopen- tanoate produced no phototoxicity and no photoallergenicity. Mutagenicity, carcinogenicity, and teratogenicity data were not available. lsostearyl Neopen- tanoate was not considered to be a significant comedogenic agent. On the basis of available data, it is concluded that this ingredient is safe as a cosmetic ingredient in its present practices of use.

CHEMISTRY

sostearyl Neopentanoate (CAS No. 58958-60-4) is the ester of isostearyl alcohol I and neopentanoic acid. It conforms to the formula:

CH, 0 I II HJI - C - C - OClBH37

This cosmetic ingredient is also known as Ceraphyl 375; Cyclochem INEO; Schercemol 85; and 2,2-dimethylpropanoic acid, isooctadecyl ester.“~‘) It is prepared by esterifying isostearyl alcohol with neopentanoic acid in the presence of a catalyst; the resulting product is purified by a proprietary process.“) Reported impurities for lsostearyl Neopentanoate include neopentanoic acid (0.4 percent maximum) and isomers of isostearyl alcohol. c3+) The chemical and physical properties of lsostearyl Neopentanoate are listed in Table 1.

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TABLE 1. Chemical and Physical Properties of lsotearyl Neopentanoate as Used in Cosmetics(‘~5.6+‘r

Appearance Clear, slightly yellow liquid Molecular weight range 348-390 UV absorption Peak absorbance in isopropyl alcohol solvent occurs at approx. 270 nm Acid value (mg KOH/g) 2.0 maximum Saponification value (mg KOH/g) 144-161 Refractive index (at 25T) 1.4485-1.4515 Pounds per gallon (at 25°C) 7.2 Cloud point -1O.O”C Iodine value 8.0 Flash point (open cup) > 180°C Free fatty acid as 0.4 percent maximum Neopentanoic Acid (mol. wt. 102) Specific gravity (at 25°C) 0.858-0.870

Solubilities (at 5 percent): Isopropyl myristate Soluble Oleyl alcohol Soluble Peanut oil Soluble Mineral oil Soluble 95 percent ethanol Soluble Propylene glycol Soluble Polypropylene glycol 3025 Insoluble Polyethylene glycol 400 Insoluble 80 percent ethanol Insoluble 70 percent sorbitol Insoluble Water Insoluble

COSMETIC USE

lsostearyl Neopentanoate is used in cosmetics for its emollient properties. It functions as a pigment-dispersing agent in eye makeup preparations and as a binder for pressed powder makeup.t6) Product formulation data submitted to the Food and Drug Administration (FDA) in 1981 by cosmetic firms participating in the voluntary cosmetic registration program indicated that lsostearyl Neopentanoate was used that year in a total of 208 cosmetic formulations (Table 2). The product type in which lsostearyl Neopentanoate was most frequently used was eye shadow (135 products). Re- ported concentrations of this ingredient in cosmetics were as follows: > 25 to 50 percent (4 products), > 10 to 25 percent (12 products), > 5 to 10 percent (119 products), > 1 to 5 percent (71 products), and > 0.1 to 1 percent (2 products).(7.8) Voluntary filing of product formulation data with the FDA by cosmetic manufacturers and formulators conforms to the prescribed format of preset concentration ranges and product categories as described in Title 21, Part 720.4 of the Code of Federal Regulations. (9) Because data are only submitted within the framework of preset concentration ranges, opportunity exists for overestimation

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TABLE 2. Product Formulation Datar’.8)

No. of Product Formulations Within Total No. of Total No. Each Concentration Range (percent)* Formulations Containing Product Category in Category ingredient >25-50 >JO-25 >5-JO >J-5 >O. J-7

lsostearyl Neopentanoate

Eyeliner 396 5 - - 5 - - Eye shadow 2582 135 - - 98 37 - Eye makeup remover 81 1 - 1 - - - Other eye makeup preparations 230 3 - - 2 1 - Blushers (all types) 819 20 2 3 8 7 - Makeup foundations 740 10 - - 2 8 - Makeup bases 831 16 2 7 - 7 - Rouges 211 2 2 - Other makeup preparations 530 - 1 - - - (not eye) Skin cleansing preparations 680 - - 1 (cold creams, lotions, liquids, and pads) Face, body, and hand 832 1 - - - 1 - skin care preparations (excluding shaving preparations) Moisturizing skin care 747 8 - - 3 4 1 preparations Night skin care preparations 219 - - - 1 - Other skin care preparations 349 - - - 1 - Suntan gels, creams, and liquids 164 - - - 2 - Other suntan preparations 28 - 1

1981 TOTALS 208 4 12 119 71 2

*Preset product categories and concentration ranges in accordance with federal filing regulations (21 CFR 720.4). W

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of the actual concentration of an ingredient in a particular product. An entry at the lowest end of a concentration range is considered the same as one entered at the highest end of that range, thus introducing the possibility of a two- to ten-fold error in the assumed ingredient concentration. Cosmetic products containing lsostearyl Neopentanoate are applied to or have the potential to come in contact with eyes and skin. Frequency and duration of application of these products will vary. Formulations incorporating Iso- stearyl Neopentanoate as an ingredient may be used as infrequently as once a week to as frequently as several times a day. Many of these products may be expected to remain in contact with the skin for as briefly as a few hours to as long as a few days. Each cosmetic product containing lsostearyl Neopentanoate has the potential for repeated application over the course of several years. A cosmetic makeup cream containing lsostearyl Neopentanoate as a component ingredient has been described in a French patent.“‘) This was the only reference to lsostearyl Neopentanoate in the published literature of which the Panel was aware.

ANIMAL TOXICOLOGY

Acute Oral Toxicity

The acute oral toxicity of cosmetic products containing various concentrations of lsostearyl Neopentanoate was assessed in both rats and mice (Table 3). In a study to evaluate 100 percent lsostearyl Neopentanoate, the acute oral LDso in rats was > 40 ml/kg.(12’

Skin Irritation

One hundred percent lsostearyl Neopentanoate caused no skin irritation in 24-hour patch tests with rabbits. (13-15)Cosmetic formulations containing various concentrations of lsostearyl Neopentanoate produced skin reactions in rabbits ranging from no skin irritation to minimal or mild skin irritation (Table 4).

Skin Sensitization

Two guinea pig studies were conducted to evaluate the skin sensitization ability of lsostearyl Neopentanoate. Each study employed a different test methodology. Ten albino guinea pigs were tested by the Magnusson-Kligman maximization procedure(16) to determine the skin sensitization potential of lsostearyl Neopen- tanoate in petrolatum. The procedure called for three test phases: induction, booster, and challenge. During the induction phase, the left and right upper back area of each of the 10 female guinea pigs received 0.05 ml intradermal injections of 50 percent aqueous Freund’s complete adjuvant, 5 percent lsostearyl Neopen- tanoate in propylene glycol, and 5 percent lsostearyl Neopentanoate in 50 percent aqueous Freund’s complete adjuvant. Twenty-four hours before the booster, 10 percent aqueous sodium lauryl sulfate was applied unoccluded to the induction site. One week after the induction injection, a 0.1 ml topical booster of “full- strength” lsostearyl Neopentanoate was applied under an occlusive dressing to the same test site as used in the induction. The booster site remained occluded

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FINAL REPORT: SAFETY ASSESSMENT OF ISOSTEARYL NEOPENTANOATE 5 for 48 hours. Two weeks following the topical booster, each guinea pig was challenged with 0.1 ml of 1 .O percent lsostearyl Neopentanoate in petrolatum on previously untreated sites on the left or right flank. The challenge dose was kept under an occlusive patch for 24 hours, after which time the patch was removed and the test site graded. One guinea pig developed barely perceptible to minimal skin erythema 48 hours following removal of the occlusive challenge patch, whereas a second animal developed the same reaction 72 hours following removal of the challenge patch. A third guinea pig had a barely perceptible to minimal skin erythema at both the 48- and 72-hour challenge evaluations. However, the reactions of this guinea pig may not have been due to lsostearyl Neopentanoate because it appeared that the animal may have scratched the test site. One of six control guinea pigs also developed minimal skin erythema. It was the investigator’s opinion that lsostearyl Neopentanoate “did not demonstrate any discernible potential for allergic skin sensitization.“(31) The skin sensitization potential of lsostearyl Neopentanoate in pyrogen-free physiological saline was determined in 10 male, albino guinea pigs by the Land- Steiner and Jacob method. (32) The test material was injected intracutaneously into the shaved back 3 times a week until a total of 10 injections had been made. The first injection for each animal consisted of 0.05 ml, and the remaining 9 injections were 0.1 ml each. Two weeks after the tenth injection, a challenge (retest) dose consisting of 0.05 ml of a freshly prepared solution was administered intracutaneously. The eleventh (retest) injection was administered just below the region of the 10 sensitizing injections. Twenty-four hours following each dose, skin reactions were evaluated and scored for diameter, height, and redness. The score for each animal was 0.0. The investigator considered lsostearyl Neopenta- noate in physiological saline a “nonsensitizer.“(33’

Comedogenicity/Pustologenicity

lsostearyl Neopentanoate was tested in two comedogenicity assays. In one assay, the ingredient at both 100 percent concentration and 50 percent concentration in mineral oil was applied to the ears of albino rabbits 5 days a week for 4 consecutive weeks (20 applications). One hundred percent lsostearyl Neopenta- noate was not comedogenic, whereas 50 percent lsostearyl Neopentanoate in mineral oil was marginally comedogenic; neither of these materials was pustulo- genic.C34) In the second assay, a night cream containing 3 percent lsostearyl Neopenta- noate was evaluated by application of the formulation to the ears of albino rabbits 5 days a week for 5 consecutive weeks (25 applications). The cream produced no significant comedone formation.tJ5)

Eye Irritation lsostearyl Neopentanoate at 100 percent concentration was at most a minimal irritant to the rabbit eye. (14*36)Ocular irritation to cosmetic products containing this ingredient varied according to the formulation tested; eye reactions in rabbits ranged from no irritation to minimal irritation. Results of these studies are summarized in Table 5.

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TABLE 3. Acute Oral Toxicity

lsostearyl Neopentanoate Concentration No. and Kind Single Oral Material Tested (percent) of Animal Dose LD,, of Material Comments Reference

lsostearyi Neopentanoate 100 5 groups of Wistar 2.5, 5.0, 10.0, >40 ml/kg No deaths during 14-day 12 albino rats 20.0, or observation period (5 rats/group) 40.0 ml/kg following the single oral dose Face makeup foundation 18 5 Sprague-Dawley 15.8 g/kg > 15.8 g/kg 17 (containing 36 percent rats (product in lsostearyl Neopentanoate) corn oil) as a 50 percent suspension in corn oil Lip product 16.05 10 fasted Sprague- 20 ml/kg >20 ml/kg No abnormal findings 18 Dawley rats (5 M (product) observed at necropsy, and 5 F) which was performed 14 days after the single oral dose Cosmetic product (con- 12.5 3 groups of 5, 10, or > 20 g/kg No deaths or abnormal 19 taining 25 percent Iso- Sprague-Dawley 20 g/kg (product in behavioral reactions dur- stearyl Neopentanoate) rats (6 rats/group) sesame oil) ing 2 weeks following the as a 50 percent suspen- single oral dose; no lesions sion in sesame oil found at necropsy

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Blusher (containing 32 8 10 fasted Harlan 5 de .5 glkg Poor grooming and soft 20 percent lsostearyl Neo- Wistar rats (5 M (product in stools observed for 3 days pentanoate) as a 25 per- and 5 F) corn oil) after the single oral dose. cent suspension in Males had an average corn oil weight loss of 25 g over a T-day period, whereas females gained an average of 37 g over same period; no deaths occurred Night cream (containing 3 1.5 10 Sprague-Dawley 5 g/kg >5.0 g/kg No deaths during 14day 21 percent lsostearyl Neo- rats (5 M and 5 F) (aqueous product observation period follow- pentanoate) as a 50 solution) ing the single oral dose percent (w/v) aqueous solution Moisturizing lotion 1.5 5 Sprague-Dawley 15.9 g/kg >15.9 g/kg - 22 2 rats (product) 5i Body oil 2.5 10 Charles River 15 ml/kg > 15 ml/kg No deaths during 5-day 23 0 3 CF-1 mice (product) observation period following the single oral dose

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TABLE 4. Skin Irritation

lsostearyl Neopentanoate No. of Material Tested Concentration (percent) Albino Rabbits Methods Comments/Results Reference lsostearyl Neopentanoate 100 3 Draize et al.‘? No skin irritation observed 13 0.5 ml of test material at the 24- or 72-hour applied under occlusive evaluations 24-hour patch to intact and abraded skin lsostearyl Neopentanoate 100 9 0.5 ml of test material No skin irritation observed 14 applied under occlusive at the 24- or 72-hour 24-hour patch to clipped evaluations skin of the back lsostearyl Neopentanoate 100 0.5 ml of test material No skin irritation observed 15 applied under occlusive at the 24- or 72-hour 24-hour patch to clipped evaluations skin of the back lsostearyl Neopentanoate 30 Test material applied No skin irritation observed 15 in aqueous solution under occlusive 24-hour at the 24- or 72-hour patch to the clipped skin evaluations Blusher 32 3 0.5 ml of product applied Slight edema and dehydration 20 daily for 4 days to of skin observed on Days 6 shaved back and 7 of a 7-day observation period; irritation index was 0.3 on a scale of 0 to 8.0

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Lip product 16.05 6 0.5 ml of product applied 4/6 rabbits had a very slight 25 under open patch to skin erythema at the 72-hour intact, clipped skin daily evaluation for 3 days Moisturizer 5 9 Product applied under Irritation scores at the 2- 26 occlusive dressing to and 24-hour evaluations were clipped skin for unspe- 0.39 and 0.06, respectively; cified period of time the Primary Irritation Index was 0.39 on a scale of 0 to 4, indicating barely perceptible to minimal skin erythema Night cream 6 48-hour patch containing The mean primary irritation 27 product applied to score was 2.67, indicating clipped intact skin mild irritation of back Body oil 2.5 3 Draize? 0.5 ml of No skin irritation observed 29 product applied under at the 24- or 72-hour occlusive 24-hour patch evaluations to intact and abraded skin Eye shadow (aqueous 1.2 9 Product applied under No skin irritation observed 30 “slurry) closed patch to clipped at the 2- or 24-hour skin for unspecified evaluations period of time

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TABLE 5. Eye Irritation

lsostearyl Neopentanoate No. of Treatment of Material Jested Concentration (percent) Albino Rabbits Exposed Eyes * Comments/Results Reference lsostearyl Neopentanoate 100 6 NWR The average score was 1 .O, 1 .O, 14 and 0.0 on Days 1, 2, and 3 postinstillation, respectively (max. score/observation - 110); investigators considered the eye irritation to be “minimal” lsostearyl Neopentanoate 100 3 NWR Slight movement of nictitating 36 membrane observed immediately following test material exposure; no irritation 1, 2, or 3 days post- instillation; behavior patterns and eating habits “within normal limits” during the 3-day observation period Face makeup foundation 36 3 TNS No ocular irritation observed 17 Blusher 32 6 TNS Slight conjunctival redness 20 observed in each rabbit 1 hour postinstillation; however, conjunctivae were clear at the 24 and 48 hour evaluations; cornea1 and iridial membranes appeared normal throughout the 7-day observation period Lip product 16.05 6 NWR Z/6 rabbits had conjunctival irri- 38 tation 24 hours posttreatment; however, no irritation was observed at the 72-hour grading; the product was not considered an eye irritant under Federal Hazardous Substances Act regulations”‘)

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Moisturizer 5 6 NWR 116 rabbits had minimal eye irri- 26 tation 24 hours postinstillation; however, no irritation was observed at the 48-hour evaluation Moisturizer NWR No ocular irritation observed 39 1, 2, 3, 4, or 7 days post- instillation Night cream WRf3 rabbits) No ocular irritation observed in 40 4 seconds rabbits given a water rinse; in after expo- the group receiving no water sure; NWR rinse, 2/6 rabbits had conjunc- (6 rabbits) tival irritation at the 24-hour evaluation; no irritation was observed 2, 3, or 7 days post- instillation; the product was considered “practically nonirritating” to the no-rinse groups Body oil 2.5 3 NWR No ocular irritation observed 41 1, 2, 3, 4, or 7 days post- instillation Moisturizing lotion 1.5 3 TNS Minimal conjunctival irritation 22 observed Eye shadow 1.2 6 NWR Z/6 rabbits developed minimal 30 ocular irritation

*In each study, the test material was instilled into one eye of the rabbit; the untreated eye served as a control. The single dose for liquid test materials was 0.1 ml. In some instances following the single exposure, the treated eye received a water rinse to remove residual test material. WR, water rinse; NWR, no water rinse; INS, treatment not specified.

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12 COSMETIC INGREDIENT REVIEW

Phototoxicity

A night cream containing 3 percent lsostearyl Neopentanoate was evaluated for its ability to cause phototoxicity. Gauze patches containing the product were applied under occlusion to both the left and right sides of the clipped back of each of 6 New Zealand rabbits. No abrasions were made to the skin. After 2 hours of exposure, patches on the right side were removed and the test sites subsequently irradiated with ultraviolet (UV) light (Sylvania Light No. F-40-BLB) for 15 minutes. The patches on the irradiated side were then replaced. Forty-eight hours following UV exposure, all patches were removed. Treated sites were graded for erythema and edema at 49, 72, and 96 hours post-UV exposure. The mean primary skin irritation score calculated for nonirradiated sites and the mean phototoxic irritation score calculated for irradiated sites were both 2.67, indicating mild skin irritation. No significant difference was observed between nonirradiated and irradiated sites. It was concluded that the product containing 3 percent lsostearyl Neopentanoate was a mild primary skin irritant, but not a phototoxic skin irritant.t2’)

Subchronic Oral Toxicity

lsostearyl Neopentanoate (100 percent) was administered by gavage to Sprague-Dawley rats for 93 days. Four groups of animals consisting of 10 males and 10 females per group were given the test material in doses of 0, 1 .O, 2.0, or 4.0 ml/kg. All animals survived the duration of the study, and no adverse behavioral responses were noted. Weekly mean body weights and total body weight gains for all treatment groups were comparable to control values. “Unthrifty” hair was observed in each of the three exposed groups, and matted and oily hair occurred in groups receiving the 2.0 and 4.0 ml/kgdosage regimen. Hematocrit values and mean corpuscular volume were increased in males of the low dose (1 .O ml/kg) group at Week 7. Increases were observed in the neutrophil:lymph ratio at Week 13 in males of the high dose (4.0 ml/kg) group. Changes in serum glucose, serum alkaline phosphatase, serum urea nitrogen, urinary protein, urine specific gravity, and urinary pH were also noted in the three treatment groups. These changes were considered of no toxicological importance, since they were within the range of normal values established for the strain of animal used. In the case of serum glucose, the changes were also considered of no toxicological significance, since they were neither unidirectional nor time- or dose-related. Other hematologic parameters (hemoglobin, total and differential leukocyte counts, red blood cell count, mean corpuscular hemoglobin concentration), clinical chemistry parameters (serum glutamic pyruvic transaminase, serum glutamic oxaloacetic transaminase), and components of the urinalysis (glucose, occult blood, bilirubin, keytones, color) for each of the treatment groups were comparable to those of the control group. In the high dose (4.0 ml/kg) group, significant increases were observed in weights of liver and heart. The increase in hepatic weight was associated with a slight increase in the frequency and severity of hepatic cytoplasmic vacuolation; however, these changes were not considered pathologically significant. Renal weights were increased and splenic weights were decreased in the low dose (1 .O ml/kg) group, but there were no histopathological changes in these organs. No significant differences were observed between exposed and control groups with respect to weight of adrenals, brain,

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FINAL REPORT: SAFETY ASSESSMENT OF ISOSTEARYL NEOPENTANOATE 13 lung, testes, or uterus. Gross examination of the skin, lungs, eyes, testes, liver, and cecum revealed no treatment-related lesions. Microscopic examination of the heart, spleen, stomach, lung, liver, brain, kidneys, adrenals, pancreas, testes, uterus, and bone marrow (sternum) also revealed no changes that could be attributed to the oral administration of lsostearyl Neopentanoate. The investigators concluded that lsostearyl Neopentanoate is safe in terms of cumulative systemic toxicity.“’

CLINICAL ASSESSMENT OF SAFETY

Skin Irritation and Sensitization

The ability of lsostearyl Neopentanoate or cosmetic products containing this ingredient to cause skin irritation and sensitization was assessed in a number of clinical studies. One hundred percent lsostearyl Neopentanoate induced no skin sensitization and no significant skin irritation. Skin irritation to products containing lsostearyl Neopentanoate varied according to test methodology and product but was generally mild at most. Cosmetic products containing this ingredient produced no skin sensitization. Individual studies are discussed below; results are summarized in Table 6. Ten subjects were exposed to 100 percent lsostearyl Neopentanoate in a 48- hour patch test. The upper portion of the back was used as the site of test material (0.5 ml) application. Skin irritation was graded on a scale of 0 (no reaction) to 6 (erythema with infiltration, vesicles, pustules, and/or erosions) 2 hours following removal of the patch. The test material elicited a skin reaction in 1 of the 10 subjects. This individual’s reaction was given a score of 1 .O, indicating a “slight noninflammatory change in surface structure.“(42) Two 48-hour patch tests were conducted to evaluate the skin irritating ability of a night cream containing 3 percent lsostearyl Neopentanoate. The site of test material contact was the upper back in one study (10 subjects) and the forearm in the second study (10 subjects). Skin response was graded 2 hours following removal of the patches. No irritation was observed in the 20 subjects tested.(43,44) No skin irritation was observed in another 48-hour patch test when 100 women were exposed on the upper back to a moisturizer containing 5 percent lsostearyl Neopentanoate. (45) In this test, treated sites were graded for erythema and edema both 15 minutes and 24 hours following removal of the occlusive patch. Cosmetic products containing lsostearyl Neopentanoate were evaluated for skin irritation in three 24-hour patch tests. In the first study, 2 of 20 individuals developed skin irritation when given a single application of a moisturizer containing 5 percent lsostearyl Neopentanoate. Of the 2 reactors, one demonstrated a “barely perceptible” to minimal skin erythema, whereas a second had mild skin erythema. The Primary Irritation Index was 0.08, indicating that the moisturizer was a minimal skin irritant.(46) A lotion containing 4 percent lsostearyl Neopenta- noate was also a minimal skin irritant (PII = 0.08) in a second 24-hour patch test involving 20 subjects. Two individuals developed “barely perceptible” to mild skin erythema in this study as well. (47) No skin irritation was observed in a third study when another 20 subjects were exposed for 24 hours to an eye shadow formulation containing 1.2 percent lsostearyl Neopentanoate.(48’

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TABLE 6. Clinical Skin Irritation and Sensitization

Isostearyl Neopentanoate Concentration No. of Test Material Tested (percent) Subjects Method Comments/Results Reference

Skin irritation lsostearyl Neopentanoate 100 10 Single 48-hour patch l/l 0 subjects developed a 42 “slight noninflammatory change in surface structure” Skin irritation Night cream 10 Single 48-hour patch No skin irritation 44 (applied to back) Skin irritation Night cream 10 Single 48-hour patch No skin irritation 43 (applied to forearm) Skin irritation Moisturizer 100 Single 48-hour patch No skin irritation 45 Skin irritation Moisturizer 20 Single 24-hour patch 2/20 subjects developed 46 “barely perceptible” to mild skin erythema (Pll = 0.08) Skin irritation Lotion 4 20 Single 24-hour patch 2/20 subjects developed 47 “barely perceptible” to mild skin erythema (PII = 0.08) Skin irritation Eye shadow 1.2 20 No skin irritation 48 Skin irritation Face makeup foundation 36 101 Fisher”” - “Negative” 17 Skin irritation Moisturizing lotion 1.5 100 Fisher”” “Negative” 50 Skin irritation Moisturizer 5 20 “Use Test!’ l/20 subjects developed 51 (3 weeks of dryness and/or increased product use) scaliness of skin Skin irritation Cream 3 15 Daily patches to same Mild skin irritation 52 site for 21 consecutive days Skin irritation/ lsostearyl Neopentanoate 100 52 Modified Draize- No skin sensitization and 53 sensitization Shelanski repeat no significant skin insult patch test irntatlon

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Skin irritation/ Raw material mixture of 60 103 Repeat insult patch Minimal skin irritation; 54 sensitization 40 percent mineral oil test no skin sensitization, and 60 percent Iso- hypopigmentation, or stearyl Neopentanoate hyperpigmentation observed Skin irritation/ Blusher 32 210 Shelanski-Jordan 2/210 subjects developed 55 sensitization repeat insult patch skin irritation (erythema test and papules); no skin sensitization was observed Skin irritation/ Blusher 32 151 Modified Draize- No skin irritation or 56 sensitization Shelanski repeat sensitization insult patch test Skin irritation/ Face product 25 54 Modified Draize- No skin irritation or 57 sensitization Shelanski repeat sensitization insult patch test Skin irritation/ Lip product 16.05 198 Modified Draize- 4/l 98 subjects developed 58 sensitization Shelanski repeat skin irritation (macular, insult patch test faint erythema); no sensitization was observed Skin irritation/ Moisturizer 5 107 Repeat insult patch 2/107 subjects developed 59 sensitization test minimal to mild skin erythema during induction phase; no skin sensitization was observed Skin irritation/ Moisturizer 5 103 Repeat insult patch 6/l 06 subjects developed 60 sensitization test skin irritation during induction phase; no skin sensitization was observed in 98 panelists completing the study

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16 COSMETIC INGREDIENT REVIEW

A face makeup foundation containing 36 percent lsostearyl Neopentanoate and a moisturizing lotion containing 1.5 percent of the ingredient were tested for their skin-irritating properties on 101 and 100 subjects, respectively. The methods employed in each study were similar to those described by Fisher.(4g’ Occlusive patch test results for both products were reported as “negative.“(17~50) A “use test” was conducted with a moisturizer containing 5 percent lsostearyl Neopentanoate on two groups of teenagers (20 subjects/group). One group used the test moisturizer for 3 weeks followed by use of control moisturizer for the same length of time. The second group used the same two products but in reverse order. Skin conditions were assessed before the test and after each 3 weeks of product use. One of 20 subjects developed dryness and/or increased scaliness of the skin as a result of the test moisturizer, whereas no subjects had skin reactions to the control moisturizer. The investigator concluded that the test moisturizer’s “capacity for evoking irritation is considered to be no different from that which would result from comparable usage of a currently-marketed moisturizer.“(51) A cosmetic cream containing 3 percent lsostearyl Neopentanoate was tested for,cumulative skin irritation. Patch applications of the product were made to the upper back on the same site daily for 21 consecutive days. Mild irritation was observed in the 15 female volunteers completing the study.(5z) A modified Draize-Shelanski patch test was conducted to evaluate the ability of 100 percent lsostearyl Neopentanoate to cause primary skin irritation and/or skin sensitization. The cosmetic ingredient was applied under occlusion for 10 24-hour periods to the same site on the back of 43 female and 9 male subjects. After a 12-day nontreatment period, a 48-hour challenge patch was applied to the back. A second 48-hour challenge patch was applied following removal of the initial challenge patch. The challenge sites were graded both immediately and 24 hours after removal of each patch. Undiluted lsostearyl Neopentanoate caused no sensitization and no significant irritation.(53) A repeated insult patch test was conducted on 103 female Caucasian subjects to assess skin irritation and sensitization of a raw material containing 40 percent mineral oil and 60 percent lsostearyl Neopentanoate. Mineral oil was also included in the testing as a nonirritating control. Patches containing the test substances were applied under semi-occlusion to the intact skin of the upper back of each subject. The patches remained in place for 48 hours (72 hours on week- ends) and then removed. The treated sites were then graded and new patches were applied. This procedure was repeated for a total of 10 induction applications. Following a 2-week nonexposure period, a challenge patch was applied. The mean irritation scores were 0.117 f 0.042 and 0.320 f 0.151 for the nonirritating control (mineral oil) and the mineral oil/lsostearyl Neopentanoate mixture, respectively (the grading scale was not specified). No skin sensitization, hypopigmentation, hyperpigmentation, or other adverse reaction were observed.(54) A Shelanski-Jordan Repeat Insult Patch Test was conducted on 210 subjects to determine the skin irritation and sensitization potential of a blusher containing 32 percent lsostearyl Neopentanoate. Test subjects consisted of both males and females between the ages of 18 and 65. The test material was placed on a gauze dressing and applied to the upper back of each subject for 24 hours. Following removal of the patch, test sites were graded for erythema and edema. Scores for

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FINAL REPORT: SAFETY ASSESSMENT OF ISOSTEARYL NEOPENTANOATE 17

skin reactions were based on a scale of 0 (no irritation) to 4.0 (marked edema and vesicles). This test procedure was repeated every other day (Monday, Wednes- day, Friday) for 3’/2 weeks for a total of 10 induction applications. Ten to 14 days after grading the tenth induction application, a challenge patch was applied for a 48-hour contact period. Seven to 10 days after removal of the initial challenge patch, a second 48-hour occlusive challenge patch was applied. Test sites were graded 48 hours following removal of the first challenge patch, and both 48 and 72 hours following removal of the second challenge patch. Two of the 210 subjects had single 2+ (erythema and papules) induction reactions; one reaction occurred after induction application number 6 and the second after induction application number 9. According to the investigator, these two reactions appeared to be “nonspecific irritation.” There were no reactions to the two challenge applications. It was concluded that the blusher containing 32 percent lsostearyl Neo- pentanoate was “neither a strong irritant nor a contact sensitizer.“(55) The skin irritation and sensitization potential of a blusher containing 32 percent lsostearyl Neopentanoate was evaluated by means of a modified Draize- Shelanski Repeat Insult Patch Test. The product was applied under an occlusive patch to the upper back of each of 151 subjects. After 48 hours, the patch was removed and the test site subsequently graded for erythema and edema. This test procedure was repeated every other day (Monday, Wednesday, Friday) for 3% weeks for a total of 10 induction applications. Ten to 14 days after the tenth insult, a challenge patch containing the test substance was applied for 48 hours. No skin reactions were observed on any of the subjects throughout the entire patch series. (56) A face product containing 25 percent lsostearyl Neopentanoate produced no skin irritation or sensitization when tested on 43 female and 11 male subjects in a modified Draize-Shelanski-Jordan patch test. The procedure called for a total of 10 24hour induction applications of the product to the upper back of each subject. After a 12-day nontreatment period, a challenge patch containing the product was applied for 48 hours. A second 48-hour patch was applied 7 days later. Challenge sites were evaluated 48 and 72 hours following application. All applications of the cosmetic product were under occluded conditions.(57J One hundred ninety-eight male and female subjects between the ages of 16 and 60 participated in a modified Draize-Shelanski patch test. A lip product containing 16.05 percent lsostearyl Neopentanoate was impregnated onto impermeable patches, which were then applied to the upper back on Monday, Wednes- day, and Friday for 3 consecutive weeks. The patches were removed and the test sites evaluated on the next scheduled patch replacement day. At the conclusion of this induction phase, a 2-week nontreatment period ensued, followed by two consecutive 48-hour challenge patches to previously untreated sites on the upper back. Challenge sites were graded at 48 and 96 hours. Skin reactions were scored on a scale of 0 (no reaction) to 4+ (bullae or extensive erosions). Four of the 198 subjects had single 1 + reactions (macular, faint erythema involving at least 25 percent of the test area); these reactions occurred following induction applications 2, 4, 7, and 8. No skin reactions occurred as a result of the challenge patches. It was concluded that the lip product had no clinically significant potential for primary irritation or contact sensitization.‘58) One hundred seven panelists were tested with a moisturizer containing 5 percent lsostearyl Neopentanoate to determine the product’s ability to cause skin

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18 COSMETIC INGREDIENT REVIEW irritation and/or sensitization. The induction phase consisted of a 24hour occlusive patch to the upper back every other day for 3 consecutive weeks (nine 0.1 ml applications). A 24-hour occlusive challenge patch was applied in the sixth week of the study. Two of the 107 subjects developed skin reactions. Of the two reactors, one developed minimal to mild skin erythema after induction applications 2 through 8. The second reactor had moderate skin erythema as a result of the second induction patch. The latter subject’s response was reported as indicative of “presensitization,” that is, her intolerance existed before the test. No other panelists developed reactions during the induction or challenge phase. It was concluded that the moisturizer “did not exhibit any potential for inducing allergic sensitization.“(59) A moisturizer containing 5 percent lsostearyl Neopentanoate was evaluated for its skin irritating and sensitizing properties on 103 women between the ages of 18 and 65. The moisturizer was applied under an occlusive dressing to the upper back for a 48-hour contact period. Upon removal of the patch, the treated site was wiped free of excess moisturizer and graded for irritation. This procedure was repeated for a total of 10 applications with the exception that “. . . patches applied on Friday remained in place for 72 hours instead of 48 hours.” After a lo-day nonexposure period, a 48-hour challenge patch was applied to a fresh site of the back. The treated site was graded both 15 minutes and 24 hours following patch removal. Six of the 103 panelists developed a positive irritation reaction during the induction period. The maximum reaction of these 6 individuals consisted of 5 “doubtful” reactions and 1 “erythema” reaction. None of the 98 panelists completing the challenge phase (5 subjects withdrew from the study during the challenge phase) had reactions to the challenge patch. Of the 5 subjects who withdrew from the test during the challenge phase, 2 had reactions during the induction period.f60)

Photosensitization A lip formulation containing 16.05 percent lsostearyl Neopentanoate was evaluated for its ability to induce photosensitization. A 0.1 ml/cm’ dose of the product was applied under an occlusive dressing to the skin of 27 subjects. After 24 hours of exposure, the patches were removed and the test sites subsequently irradiated with a UV light source at three times the individual’s minimal erythema dose (MED). The MED of each panelist was determined in accordance with procedures outlined in the Federal Register. (61) A filtered Xenon Arc Solar Simulator (150 W) was used to produce a continuous emission spectrum in the UVA and UVB region (290-400 nm). Forty-eight hours following the UV exposure, the test sites were graded. The procedure of application, patching, and UV treatment was then repeated for a total of seven product and UV exposures. No phototoxicity or photoallergenicity was observed in any subject.(6z)

SUMMARY

lsostearyl Neopentanoate is the ester of isotearyl alcohol and neopentanoic acid. It is used in cosmetics for its emollient properties. The ingredient functions as a binder for pressed powder makeup and as a pigment-dispersing agent in eye

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FINAL REPORT: SAFETY ASSESSMENT OF ISOSTEARYL NEOPENTANOATE 19

makeup formulations. Although lsostearyl Neopentanoate is used in a variety of cosmetic products, its most frequent use occurs in eye shadow. Concentrations of this ingredient in cosmetic products generally range from 1 to 10 percent, although there are a few products with higher concentrations. Cosmetic products containing lsostearyl Neopentanoate are applied to the skin and/or eye area. No significant published literature existed for lsostearyl Neopentanoate. As a result, much of the safety data available to the CIR Panel consisted of unpublished reports and studies provided by industry. lsostearyl Neopentanoate (100 percent) was nonirritating to the skin, minimally irritating to the eye, noncome- dogenic, and nonpustulogenic in studies with rabbits. The acute oral LDso in rats of 100 percent lsostearyl Neopentanoate was > 40 ml/kg, indicating that this ingredient was relatively nontoxic to this animal by oral administration. In studies with guinea pigs, 1.0 percent lsostearyl Neopentanoate in petrolatum and 0.1 percent in physiological saline were nonsensitizing to the skin. No cumulative systemic toxicity was observed in rats given 100 percent lsostearyl Neopenta- noate orally for 93 days. In clinical studies, lsostearyl Neopentanoate (100 percent) caused no sensitization and no significant irritation of the skin. A cosmetic lip product containing 16.05 percent of the ingredient produced no phototoxicity and no photoallergenicity.

DISCUSSION

The CIR Panel noted the lack of unpublished or published data on the mutagenicity, carcinogenicity, and teratogenicity of lsostearyl Neopentanoate and its acid and alcohol components. The alcohol and the acid resulting from hydrolysis of lsostearyl Neopentanoate ester would be expected to be metabolized along regular pathways and would not be expected to be converted to mutagenic, carcinogenic, and/or teratogenic metabolites.

CONCLUSION

On the basis of the available information, the Panel concludes that lsostearyl Neopentanoate is safe as a cosmetic ingredient in the present practices of use.

ACKNOWLEDGMENT

Jonathan T. Busch, Scientific Analyst and Writer, prepared the technical analysis used by the Expert Panel in developing this report.

REFERENCES

1. ESTRIN, N.F., CROSLEY, P.A., and HAYNES, C.R. (1982). CTFA Cosmetic Ingredient Dictionary, 3rd ed. Washington, DC: The Cosmetic, Toiletry and Fragrance Association, Inc. 2. CTFA. (October 1980). Submission of unpublished data by CTFA. Cosmetic ingredient chemical description: lsostearyl Neopentanoate. CTFA Code No. 2.*

*Available upon request: Administrator, Cosmetic Ingredient Review, 1110 Vermont Ave., NW, Suite 810, Washington, DC 20005. I

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20 COSMETIC INGREDIENT REVIEW

3. COSMETECH LABORATORIES, INC. (April 26, 1983). Submission of unpublished data by CTFA. Caschro- matic analysis of lsostearyl Neopentanoate and accompanying letter from David Carlen, President (Cosme- tech Lab. Inc.) to Dr. Gerald McEwen, Cosmetic, Toiletry and Fragrance Assoc.).* 4. VAN DYK and COMPANY. (April 25, 1983). Submission of unpublished data by CTFA. Letter from Ken Klein, Director of Technical Services (Van Dyk and Co.) to David Garlen, President (Cosmetech Lab. Inc.) regarding Isostearyl Neopentanoate.* 5. AVON PRODUCTS, INC. (January 31, 1983). Submission of unpublished data by CTFA. Thirteen week subchronic oral toxicity study in albino rats. The toxicological evaluation of lsostearyl Neopentanoate (R.I. Code 0187). Study Project Code AT0202. Phase II. Final Report.* 6. VAN DYK and COMPANY, INC. (January 1976). Submission of unpublished data by CTFA. Technical Bul- letin. Ceraphyl 375. CTFA Code No. 2.* 7. FOOD and DRUG ADMINISTRATION (FDA). (December 22, 1981). Cosmetic product formulation data: ingredients used in each product category. Computer printout. Washington, DC. 8. FDA. (January 5, 1982). (a) Frequency of Trade Name Ingredients in the Cosmetic Product File and (b) Fre- quency of Common, Usual, or Chemical Names in the Cosmetic Product File. Computer printout. Wash- ington, DC. 9. CFR. (Revised as of April 1, 1982). Title 21, Part 720.4. Voluntary Filing of Cosmetic Product Ingredient and Cosmetic Raw Material Composition Statement. Information requested about cosmetic products. 10. VAN DYK and COMPANY. (January 4, 1982). Submission of unpublished data by CTFA. UV Absorption Scan (210 nm to 500 nm). lsostearyl Neopentanoate (Ceraphyl 375) in Isopropyl Alcohol solvent. Concen- tration: 10,000, 2,500, 1,250 mglL/l cm. Batch No. 9305. Approx. molecular weight 348-90.* 11, BOULOGNE, J., KOULBANIS, C., and MICHELET, J. (January 2, 1982). Cosmetic composition in the form of an anhydrous cream of oily consistency. Fr. demande Patent No. 2486800. Oreal S.A. 12. FOOD and DRUG RESEARCH LABORATORIES, INC. (FDRL). (April 6, 1976). Submission of unpublished data by CTFA. Acute oral toxicity in rats. lsostearyl Neopentanoate. CTFA Code NO. 3.* 13. BIO-TOXICOLOGY LABORATORIES, INC. (BTL). (April 7, 1972). Submission of unpublished data by CTFA. Primary skin irritation in rabbits. lsostearyl Neopentanoate. CTFA Code No. 5.’ 14. CTFA. (May 20, 1980). Submission of unpublished data by CTFA. Toxicology Summary Report: Animal Tests. Skin and eye irritancy in rabbits. lsostearyl Neopentanoate. Batch No. 2985. CTFA Code NO. 4.* 15. CTFA. (May 20, 1980). Submission of unpublished data by CTFA. Toxicology Summary Report: Animal tests. Primary skin irritation in rabbits. lsostearyl Neopentanoate. Code No. 0187. CTFA Code NO. 6.* 16. MAGNUSSON, B., and KLIGMAN, A.M. (1969). The identification of contact allergens by animal assay. The guinea pig maximization test. J. Invest. Dermatol. 52(3), 268-76. 17. COSMETIC, TOILETRY and FRAGRANCE ASSOCIATION. (CTFA). (February 1977). Submission of unpub lished data by CTFA. Face makeup foundation containing 36 percent lsostearyl Neopentanoate. Acute oral toxicity. Acute eye irritation, Human patch test. In-House Code: FCR-1011011B. CTFA Code No. lo.* 18. CTFA. (May 9, 1979). Submission of unpublished data by CTFA. CIR safety data test summary response form. Acute oral toxicity in rats. Lip product containing 16.05 percent lsostearyl Neopentanoate. Company Test Code: CSE 138.’ 19. APPLIED BIOLOGICAL SCIENCES LABORATORY. (October 29, 1980). Submission of unpublished data by CTFA. Biological testing request. Acute oral toxicity in rats. Formula 2213-62-A containing 25 percent Iso- stearyl Neopentanoate in sesame oil. No. ABO-19.’ 20. CTFA. (September 8, 1978). Submission of unpublished data by CTFA. Blusher containing 32 percent Iso- stearyl Neopentanoate. Acute oral, dermal, and ocular testing of 88-006, Lot 344-06a.* 21. FDRL. (May 1, 1979). Submission of unpublished data by CTFA. Acute oral toxicity in rats. Night cream containing 3 percent Isostearyl Neopentanoate. Laboratory No. 6205e.* 22. CTFA. (February 6, 1978). Submission of unpublished data by CTFA. Moisturizing lotion containing 1.5 percent lsostearyl Neopentanoate. Acute oral toxicity. Acute eye irritation. In-house Code FCR-lO/lOl 1C. CTFA Code No. 25.’ 23. LEBERCO LABORATORIES. (October 23, 1979). Submission of unpublished data by CTFA. Acute oral toxicity in mice. Body oil containing 2.5 percent lsostearyl Neopentanoate. CTFA Code No. 22.’ 24. DRAIZE, J.H., WOODARD, G., and CALVERY, H.O. (1944). Methods for the study of irritation and toxicity of substances applied topically to the skin and mucous membrane. J. Pharmacol. Exp. Ther. 82,377-90. 25. CTFA. (May 5, 1979). Submission of unpublished data by CTFA. Skin irritation test. CIR safety data test summary response form. Lip product containing 16.05 percent lsostearyl Neopentanoate. Company Test Code: CSE 138.* 26. CTFA. (March 7, 1980). Submission of unpublished data by CTFA. Toxicology summary report: Animal tests. Skin and eye irritation in rabbits. Moisturizer containing 5 percent lsostearyl Neopentanoate. Code No. 16434-l 1. CTFA Code No. 14.*

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FINAL REPORT: SAFETY ASSESSMENT OF ISOSTEARYL NEOPENTANOATE 21

27. FDRL. (May 21, 1979). Submission of unpublished data by CTFA. Primary dermal phototoxic irritation study with New Zealand white rabbits. Laboratory No. 6164. CTFA Code No. 9.* 28. DRAIZE, J.H. (1959). Dermal toxicity in appraisal of the safety of chemicals. In: Foods, Drugs and Cosmet- ics. Div. of Pharmacology, FDA, Dept. of HEW. The Assoc. of Food and Drug Officials of the US Business Office, Bureau of Food and Drugs, Austin, Texas, pp. 46-59. 29. LEBERCG LABORATORIES. (October I 2, I 979). Submission of unpublished data by CTFA. Skin irritation in rabbits. Body oil containing 2.5 percent lsostearyl Neopentanoate. CTFA Code No. 23.” 30. CTFA. (March 31, 1980). Submission of unpublished data by CTFA. Toxicology summary report Animal test. Skin and eye irritation in rabbits. Eye shadow containing 1.2 percent lsostearyl Neopentanoate. Code No. 1823442. CTFA Code No. 27.’ 31. CTFA. (February 15, 1978). Submission of unpublished data by CTFA. Guinea pig allergy study. IsostearyJ Neopentanoate. Study Code GPA-12-78. CTFA Code Nos. 8 and 14.” 32. LANDSTEINER, K., and JACOBS, J. (1935). Sensitization of animals with simple chemical compounds. 1. Exp. Med. 61, 643. 33. BTL. (May 16, 1972). Submission of unpublished data by CTFA. Landsteiner and Jacobs guinea pig sensitization study. lsostearyl Neopentanoate.’ 34. CTFA. (September 9, 1980). Submission of unpublished data by CTFA. Comedogenic assay. Rabbit ear. Four-week assay. Samples 176-01 and 176-02. Mary Kay Cosmetics. CTFA Code No. lo.* 35. CTFA. (June 28, 1979). Submission of unpublished data by CTFA. Comedogenic assay. Rabbit ear. Night cream containing 3 percent lsostearyl Neopentanoate. CTFA Code No. 6.* 36. BTL. (April 11, 1972). Submission of unpublished data by CTFA. Eye irritation in rabbits. lsostearyl Neopen- tanoate. CTFA Code No. 7.’ 37. CODE OF FEDERAL REGULATIONS (CFR). (Revised as of April 1979). Title 16 Part 1500.42. (16 CFR 1500. 42). Federal Hazardous Substances Act Regulations. Test for eye irritants. 38. CTFA. (April 13, 1979). Submission of unpublished data by CTFA. CIR safety data test summary response form. Eye irritation test. Lip product containing 16.05 percent lsostearyl Neopentanoate. Company Test Code: CSE 138.* 39. LEBERCO LABORATORIES, INC. (October 2, 1979). Submission of unpublished data by CTFA. Eye irritation in rabbits. Moisturizer containing 5 percent lsostearyl Neopentanoate. CTFA Code No. 15.* 40. FDRL. (April 5, 1979). Submission of unpublished data by CTFA. Eye irritation test in rabbits. Night cream (light pink cream) containing 3 percent lsostearyl Neopentanoate. Laboratory No. 6205e. CTFA Code No. 11.* 41. LEBERCO LABORATORIES. (October 15, 1979). Submission of unpublished data by CTFA. Eye irritation in rabbits. Body oil containing 2.5 percent lsostearyl Neopentanoate. CTFA Code No. 24.* 42. HERNDON, J.H. (April 28, 1980). Submission of unpublished data by CTFA. Herndon: Associate Professor, Internal Medicine; Chairman, Division of Dermatology. University of Texas/Southwestern Medical School. Forty-eight hour patch test with 100 percent lsostearyl Neopentanoate. Protocol No. 001-l 67.’ 43. HERNDON, J.H. (February 27, 1979). Submission of unpublished data by CTFA. Herndon: Associate Pro- fessor, Internal Medicine; Chairman, Division of Dermatology. University of Texas/Southwestern Medical School. Forty-eight hour patch test with night cream containing 3 percent lsostearyl Neopentanoate. Proto- col No. OOl-123.* 44. HERNDON, J.H. (February 12, 1981). Submission of unpublished data by CTFA. Herndon: Associate Pro- fessor, Internal Medicine; Chairman, Division of Dermatology. University of Texas/Southwestern Medical School. Forty-eight hour patch test with night cream containing 3 percent lsostearyl Neopentanoate. Proto- col No. 001-210.’ 45. TESTKIT LABORATORIES, INC. (TKL). (June 22, 1979). Submission of unpublished data by CTFA. Human skin irritation. Forty-eight hour patch test. Moisturizer containing 5 percent lsostearyl Neopentanoate. CTFA Code No. 17.* 46. CTFA. (March 13, 1980). Submission of unpublished data by CTFA. Clinical evaluation report: Human patch test. Moisturizer containing 5 percent lsostearyl Neopentanoate. Test Material: 16434-11. CTFA Code No. 16.* 47. CTFA. (January 24, 1980). Submission of unpublished data by CTFA. Evaluation report: Human patch test. Lotion containing 4 percent lsostearyl Neopentanoate. Test Material: 17043-03. CTFA Code No. 20.* 48. CTFA. (April 10, 1980). Submission of unpublished data by CTFA. Clinical evaluation report: Human patch test. Eyeshadow containing 1.2 percent lsostearyl Neopentanoate. Test Material: 18234-02. CTFA Code No. 26.’ 49. FISHER, A.A. (1973). Contact Dermatitis, 2nd ed. Philadelphia: Lea & Febiger. 50. CTFA. (February 7, 1977). Submission of unpublished data by CTFA. Moisturizing lotion containing 1.5 percent JsostearyJ Neopentanoate. Human patch test. In-House Code FCR-lO/lOllC. CTFA Code No. 25.*

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22 COSMETIC INGREDIENT REVIEW

51. CTFA. (June 4,198O). Submission of unpublished data by CTFA. Study No. APTC-121-80. Three week product use test. Moisturizer 16434-12 containing 5 percent lsostearyl Neopentanoate.* 52. INTERNATIONAL RESEARCH SERVICES, INC. (luly, 1979). Submission of unpublished data by CTFA. Twenty-one day cumulative irritation study of fourteen products. Study 130. Cream containing 3 percent lsostearyl Neopentanoate.* 53. UNIVERSITY OF CALIFORNIA, LOS ANGELES (UCLA). Duly 19, 1977). Submission of unpublished data by CTFA. Modified Draize-Shelanski Patch Test. lsostearyl Neopentanoate (100 percent). UCLA File No. 975.* 54. CTFA. (February 26, 1982). Submission of unpublished data by CTFA. Repeat insult sensitization study. Raw material 81-81: Mineral oil (40 percent) and lsostearyl Neopentanoate (60 percent).* 55. LEO WINTER ASSOCIATES, INC. (November 24, 1980). Submission of unpublished data by CTFA. Re- peated insult patch testing. Shelanski-Jordan procedure. Blusher containing 32 percent lsostearyl Neopen- tanoate. Lot No. 344-06-A. Project-BB 006. CTFA Code No. 12.* 56. LEO WINTER ASSOCIATES, INC. (November 24, 1980). Submission of unpublished data by CTFA. Re- peated insult patch testing and in-use testing. Modified Draize-Shelanski procedure. Blusher containing 32 percent lsostearyl Neopentanoate. 88-006. CTFA Code No. 13.* 57. UCLA. (November 17, 1980). Submission of unpublished data by CTFA. Modified Draize-Shelanski-Jordan patch test. Face product containing 25 percent lsostearyl Neopentanoate. UCLA File No. 2207. Project No. H80-6-2E.’ 58. CTFA. (July/August 1979). Submission of unpublished data by CTFA. CIR safety data test summary response form. Modified Draize-Shelanski repeat insult patch test. Lip product containing 16.05 percent lsostearyl Neopentanoate. Company Test Code CSE 138.* 59. CTFA. (May 30, 1980). Submission of unpublished data by CTFA. Allergic contact sensitization test. Test No. APTC-112-80. Moisturizer 16434-l 2 containing 5 percent lsostearyl Neopentanoate. (Also titled: Con- tact allergy test report. CTFA Code No. 19).* 60. TKL. (September 7, 1980). Submission of unpublished data by CTFA. Repeated insult patch test. Moisturizer containing 5 percent lsostearyl Neopentanoate. CTFA Code No. 18.* 61. FEDERAL REGISTER. (1978). August 25, 38206-38369. 62. CTFA. Uuly/August 1979). Submission of unpublished data by CTFA. Human photosensitization study. Lip product containing 16.05 percent lsostearyl Neopentanoate. Company Test Code CSE 138.*

Supplement Panel Book 1 Page 120 Alkyl Esters Supplement Book 1 jOURNA1 OF THE AMERICAN COLLEGE OF TOXICOLOGY Volume 9, Number 2,199O Mary Ann Liebert, Inc., Publishers

Final Report on the Safety Assessmentof Arachidyl Propionate

Arachidyl Propionate is an amber-colored semisolid wax, which is used in cosmeticsat concentrations up to 10%. Arachidyl Propionate is practically nontoxic when ingested. No systemic toxicity was associated with subchronic oral exposure to Arachidyl Propionate. In test animals, Arachidyl Propionate was neither a primary dermal irritant, nor a skin sensitizer, and it did not have comedogenic potential. It was nonirritating to rabbit eyes. In clinical studies, Arachidyl Propionate was neither an irritant nor a sensitizer. In additional clinical studies, Arachidyl Propionate was neither phototoxic nor a photoallergen. On the basis of the data included in this report, it is concluded that Arachidyl Propionate is safe as a cosmetic ingredient in the present practices of use and concentration.

INTRODUCTION

rachidyl Propionate is structurally, and chemically, similar to the stearates, Aspecifically butyl stearate. The stearates have been previously reviewed,“’ and the data contained in the stearates report are applicable to the evaluation of the safety of Arachidyl Propionate as a cosmetic ingredient.

CHEMISTRY

Definition and Structure Arachidyl Propionate is the propionic acid ester of the corresponding C,, fatty alcohol. It is manufactured as a mixture of the esters of the C,,-C,, fatty alcohols, of which the C20 fatty alcohol ester is the major constituent. Arachidyl Propionate conforms to the following structure:‘2’

if CH3CH,COCH2(CH2),&H,

143

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Arachidyl Propionate (CAS No. 65591-14-2) is commercially known as Waxenol@ 801 ec2’

Properties Arachidyl Propionate is a soft waxy solid, amber in color, with a slight characteristic odor.‘2,3’ It is insoluble in waterc3’ and soluble in mineral ail.(4) It has a boiling point of 435”Fc3’and a melting range of 36-38”C.‘4’The specific gravity of Arachidyl Propionate is 0.83, the acid value is 1 .O maximum, the ester value is 100-l 16, and the iodine value is 15 maximum.(2)

Analytical Methods Ultraviolet spectral analyses have been performed on Arachidyl Propionate.‘5*6’ When diluted in cyclohexane, Arachidyl Propionate absorbed strongly in the 200-260 nm region, but, above 290 nm, absorption is poor.

CHEMICAL REACTIONS

Arachidyl Propionate is chemically stable, but will decompose upon high-temperature heating and will react with strong bases to form the parent alcohol and propionic acid.‘3’

USE

Cosmetic Arachidyl Propionate is a “specialty wax” used in cosmetic and toiletry products to provide emolliency, lubricity, gloss, and film-forming capabilities to the finished products. Arachidyl Propionate melts upon contact with the human body, leaving a nonoily feeling to the skin. Data submitted to the Food and Drug Administration in 1987 by cosmetic firms participating in their voluntary registration program indicated that Arachidyl Propionate was used in a total of 31 cosmetic products (Table 1). The cosmetic formulations containing the greatest amount of Arachidyl Propionate were lipsticks (24 formulations) with a reported concentration range of >5-10%. Other products containing Arachidyl Propionate (at concentrations of ~5%) include “other” fragrance preparations (one product), moisturizing skin care preparations, and face, body, and hand care preparations, excluding shaving preparations.“) The FDA cosmetic product formulation computer printout”’ is compiled through voluntary filing of such data in accordance with Title 21 part 720.4 of the Code of Federal Regulations. (8) Ingredients are listed in preset concentration ranges under specific product type categories. Since certain cosmetic ingredients are supplied by the manufacturer at less than 100% concentration, the value reported by the cosmetic formulator may not necessarily reflect the actual concentration found in the finished product; the actual concentration would be a fraction of that reported to the FDA. Data submitted within the framework of preset concentration ranges provides the opportu-

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TABLE 1. Product Formulation Data for Arachidyl PropionatG

No. of product formulations within each concentration Total no. of Total no. range (percent) formulations containing Product category in category ingredient >5-10 55 - Other fragrance preparations 180 1 Lipstick 1494 24 20 4 Face, body, and hand skin care 1005 3 3 preparations (excluding shaving preparations) Moisturizing skin care preparations 775 3 3

1987 Totals 31 20 11

nity for overestimation of the actual concentration of an ingredient in a particular product. An entry at the lowest end of a concentration range is considered the same as one entered as one entered at the highest end of that range, thus introducing the possibility of a two- to tenfold error in the assumed ingredient concentration. Products containing Arachidyl Propionate may be applied to the skin of all parts of the body, especially the lips, several times daily, and may be used repeatedly over a number of years.

Noncosmetic Arachidyl Propionate is used in such nonprescription over-the-counter products as ointments and emulsions.‘g’

ANIMAL TOXICOLOGY

Acute Toxicity Oral Male Wistar rats were given Arachidyl Propionate by oral intubation at 10.4, 12.8, 16.0, and 20.0 g/kg. (lo) Those rats receiving the three largest doses experienced mild diarrhea. No rats died. It was concluded that Arachidyl Propionate was not a hazardous substance under the conditions of the test. Dermal Arachidyl Propionate was tested for acute dermal toxicity in ten New Zealand White rabbits.“O’The abdominal area of the rabbits was shaved and the skin of one side was abraded while the skin of the other side remained intact. The test substance, 2.0 g/kg, was applied to the skin and the area was wrapped with gauze. The Arachidyl Propionate was allowed to remain in contact with the skin for 24 h, after which the material was removed. The rabbits were weighed on days 1 and 14, and were observed for signs of toxicity during the treatment period. There were no signs of toxicity, no deaths, and the rabbits each gained an average of 0.4 kg during the study.

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Subchronic Toxicity

Oral Groups of 20 Sprague-Dawley rats, equally divided by sex, were given Arachidyl Propionate by gavage for 5 days a week for 13 weeks. ““Therats weredivided into four dosage groups: controls received the corn oil vehicle, and three groups received 250, 750, and 2500 mg/kg, respectively, of the test substance at concentrations of 25% in corn oil. The test doses were determined in a 15day range-finding study, with the highest dose administered being limited by the amount of corn oil vehicle which could be administered without altering the serum chemistries of the rats. All rats were weighed prior to the study and weekly during the study, and doses were adjusted for any changes in body weight. The rats were observed daily for any toxicologic or pharmacologic signs, and for general appearance. Blood and urine samples were obtained for analysis during weeks 7 and 13. At the end of the study, the rats were sacrificed, and various tissues were obtained for microscopic examination. During the study, no statistically significant differences were found in the mean body weights or mean body weight gains of the rats in the experimental or control groups, and no deaths were attributed to treatment with the test material. A few rats had soft feces and some male rats had dried blood around their noses, a sign attributed to treatment. The hematologic parameters measured were percent hemoglobin values and packed cell volume, MCV, red blood cells (RBC), white blood cells (WBC), neutrophill lymphocyte ratio, and percent MCHC. Also measured were glucose, blood urea nitrogen (BUN), serum glutamic pyruvate tranaminase (SGPT), serum alkaline phosphatase (SAP), and serum glutamic oxaloacetic transaminase (SCOT). The low- and mid-dose male rats had statistically significant differences in the week 7 neutrophil/ lymphocyte ratio values when compared with the control values. There were some statistically significant differences in the hematologic parameters between female rats and their controls; however, none of the differences indicated a dose-response relationship. Some clinical chemistry values differed significantly among groups. In male rats, the SGPT and SAP values varied from those of the controls, but again no dose-response relationship was established. In female rats, differences were found in the SGOT and SAP values between control and treated rats, but there were no dose-response trends. The SAP values for the corn oil controls were higher than previously observed in that laboratory. No significant differences were found between control and experimental groups in the results of urinalyses. In the treated female rats, a trend toward a smaller heart/body weight ratio was found, and uterine weights and uterus weight/body weight ratios were decreased. A high incidence of lobular patterns in the livers of all four groups of rats was attributed to vascular congestion and was not considered a result of treatment. No microscopic lesions were found, with the exception of multifocal pneumonitis in the high-dose rats. Foamy macrophages were found in the lungs of both the control and high dose rats. The aspiration of oily substances has been considered a cause of pneumonitis, and, thus, the differences observed between the control and high dose groups were due to the combination of the corn oil and Arachidyl Propionate rather than to corn oil alone. No deaths were attributed to treatment, and no systemic toxic effects other than pneumonitis were observed. The researchers concluded that no systemic toxicity would be expected from the normal use of Arachidyl Propionate.

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Dermal Irritation A primary dermal irritation study was performed using six New Zealand White rabbits.“” The backs of the rabbits were shaved and the skin on one side of the spinal column was abraded. Arachidyl Propionate, 0.5 g, was applied to gauze squares which were then placed on the skin on both sides of the spinal column and left in place for 24 h. The skin was observed for signs of dermal irritation upon removal of the pads and again 48 h later. The Primary Irritation Index (PII) was 1.38; Arachidyl Propionate was not a primary irritant. A primary dermal irritation study (in rabbits, occlusive patch test, abraded and intact skin) with Arachidyl Propionate had a PII of 0.13 for the undiluted material.“2’ The Arachidyl Propionate tested as a 10% aqueous emulsion had a PII of 0.0. Arachidyl Propionate was classified as a nonirritant. In another dermal irritation study, a lipstick product containing 7.0% Arachidyl Propionate was tested using six rabbits (strain not specified) with a single insult, occlusive patch test. ‘13) The animals were examined for signs of irritation 2 and 24 h after removal of the patches. None of the rabbits had any signs of irritation; the group PII was 0.00, and the lipstick containing Arachidyl Propionate was classified as nonirritating. In another study performed by Guillot et al., (12)the cumulative dermal irritation potential of Arachidyl Propionate was determined. Three New Zealand White rabbits had their backs and flanks clipped; the clipping was repeated once weekly as necessary. The test substances, undiluted Arachidyl Propionate and a 10-l 5% emulsion of Arachidyl Propionate, were applied to the right and left flanks, respectively, of each rabbit. The test substance was massaged into the skin and any excess was wiped off. This procedure was repeated 5 days per week for 8 weeks. The animals were weighed weekly, and the skin was evaluated daily, with the daily scores expressed as a weekly average. At the end of the 8-week period, the skin was not treated for 7 days, and was then re-evaluated; a challenge assay was then performed. Also at the end of the treatment period, two skin samples (from areas differing macroscopically) were obtained for histopathological evaluation. For the pure test material, the Mean Maximum Irritation Index (MMII) was 1 .OO. This score classifies Arachidyl Propionate as a slight irritant. Additional observations were “slight epidermal exfoliation” and upon histopathological evaluation, the findings were “slight congestion of the dermis” with “no pathological reaction.” The MMII for the 10% emulsion of Arachidyl Propionate was 0.50, a score reflective of a nonirritant. The emulsion was well tolerated, though vesicles were observed in two rabbits. Results of histopathological evaluation were no lesions due to treatment. Arachidyl Propionate was well tolerated by the rabbit and did not appear to have allergic potential.

Dermal Sensitization Twenty male albino guinea pigs were used in a dermal sensitization study.“” The skin in the area of sample application was shaved. Ten of the animals served as controls, receiving injections of saline instead of warm, undiluted Arachidyl Propionate. A total of ten injections were administered, the first was a volume of 0.05 ml, and the remaining nine were volumes of 0.1 ml. The animals were treated three times per week, and the sites were scored 24 h after each injection. There was a 2-week nontreatment period after the tenth injection. At the end of this period, a 0.05 ml dose of the test

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148 COSMETIC INGREDIENT REVIEW

substance or saline was administered below the site of the original injections. This challenge site was scored 24 h later. No reaction was observed in any of the guinea pigs during the treatment or challenge phases of the test. The researchers concluded that Arachidyl Propionate was not a skin sensitizer.

Comedogenicity Three male albino rabbits, free from evidence of ear mites, were used in a comedogenicity assay of Arachidyl Propionate.(14’ Undiluted Arachidyl Propionate, 0.1 ml, was applied daily to the internal base of the right ear of each animal on the weekdays of three weeks (15 applications); the left external ear served as the control. Twenty-four hours after each application, the rabbits’ external ears were examined for signs of comedogenicity. The animals were examined, weighed, and sacrificed 24 hours after the last application. Necropsy was performed and histopathological observations were made of the collected tissues. Two of the rabbits had lost weight during the study, while the third had normal weight gain. Diarrhea, reduced passage of feces, and emaciation were noted (rabbits not specified) during the observation period. Observation during treatment indicated hyperkeratosis with possible comedone formation in all three rabbits. Follicular hyperkeratosis was observed in the treated ears, but not comedone formation. Hyper- keratosis was also observed in the skin of the untreated ears. The researchers concluded that Arachidyl Propionate did not have comedogenic potential.

Ocular Irritation An ocular irritation test of Arachidyl Propionate was performed using six New Zealand White rabbits.“” Warm Arachidyl Propionate, 0.1 ml, was instilled into the conjunctival sac of one eye of each rabbit; the contralateral eye served as the control. The ocular reaction was scored 24, 48, and 72 h after instillation of the test material. Two of the rabbits had conjunctival redness and discharge on day 1, but these signs had cleared by day 2. One rabbit had redness, chemosis, and discharge on day 1, persisting through days 2 and 3 (with the exception of the chemosis which was not noted on day 3). A fourth rabbit had redness, chemosis, and discharge on day 1, with the chemosis subsiding on day 2 and the discharge no longer evident on day 3. The remaining two rabbits had no reactions. Arachidyl Propionate was not an ocular irritant under the conditions of the study. A study in rabbits using official French testing procedures was performed to determine the ocular irritation index for a number of cosmetic raw ingredients, including Arachidyl Propionate. C4)Several modifications of the official test procedures included: the inclusion of a reading of the eye at 1 h in addition to the readings at 1,2, 3, 4, and 7 days. A photomotor reflex study was done, fluorescein dye was used, a qualitative evaluation of ulceration and granulation was performed, an ophthalmo- scope and a retinograph were used, and the results were scored on a scale of 1 to 100. The Ocular Irritation Index (011)for Arachidyl Propionate was 3.83 at 1 h, 1.50 at 24 h, and 0.00 at 48 h. In another study, a lipstick containing 7.0% Arachidyl Propionate was tested for ocular irritation using six rabbits. (13)The product was not rinsed from the conjunctival sac of the eyes, and the eyes were graded 1 and 2 days after the instillation. None of the

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rabbits had reactions to the product. The lipstickcontaining 7.0% Arachidyl Propionate was classified a nonirritant according to the Draize classification of ocular irritation.

CLINICAL ASSESSMENT OF SAFETY

Dermal Irritation and Sensitization A repeated insult patch test of a lipstick containing 7.0% Arachidyl Propionate was performed using a panel of 85 human subjects. (15)The study group consisted of 82 females and 3 males between the ages of 18 and 70. Approximately 0.1 ml of the test material was applied to an occlusive patch, which was then placed on the upper back of the test subject. The patches were applied every Monday, Wednesday, and Friday for 3 weeks, and remained in place for 24 h. The test sites were scored before the application of each new patch and 24 h after the removal of the last patch. After a 2-week nontreatment period, a challenge patch, which remained in place for 24 h, was applied to a previously untreated site on the back of the test subjects. The sites were scored 24 and 48 h after removal of the patch. None of the panelists had reactions during either the induction or the challenge phases of the test. The lipstick containing 7.0% Arachidyl Propionate did not have a sensitization potential under the conditions of the study. A lipstick containing 7.0% Arachidyl Propionate was tested in a 4-day mini-cum assay (a repeat insult test). (16)The test material had an irritation score of 0 in 20 test subjects (test subjects not identified), resulting in a PII of 0 and the conclusion that the lipstick containing 7.0% Arachidyl Propionate was essentially nonirritating. A lipstick containing 7.0% Arachidyl Propionate was tested in use by a group of 62 women.(“) The women were divided into two groups balanced by product use, skin type, and skin sensitivity. The women were required to apply the lipstick at least twice daily. During the first three weeks of the study, one group used the test lipstick while the second group used a control lipstick-a product that was already “in-line.” After 3 weeks, the products were crossed over; in other words, the panelists originally using the test lipstick were then using the “in-line” lipstick. The panelists’ lips were evaluated dermatologically at the beginning of the study, at the crossover date, and at the end of the study. A summary of the clinical test results was not given. No dermatologic changes were noted in any of the panelists lips during the study. One panelist noted “discomfort” with the test lipstick, but no further details were given. One panelist complained of “dry feeling,” but it was not clear if this was the same panelist who noted discomfort. The table of panelist comments and problems was not included in the report. The results of the study of the lipstick containing 7.0% Arachidyl Propionate were acceptable when compared with the in-line (control) product; the reactivity of the test product was very low. The irritation potential of a lip makeup containing 10% Arachidyl Propionate was evaluated in a supervised usage test using 30 subjects aged 29-66 years.“8’ The test subjects were evaluated for signs of dermatitis or allergic reactions after they had used the lip makeup under normal conditions for 4 weeks. The test subjects had no complaints about the lip makeup, and no evidence of dermatitis or other allergic reactions were noted. A 21-day cumulative irritation test on a lip makeup containing 10% Arachidyl Propionate was performed using 25 human subjects between the ages of 18 and 65.“”

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The lip makeup was tested along with other makeup products, and with sodium lauryl sulfate (SLS)in petrolatum at concentrations of 0.5 and 4.0% as standards (4.0% SLS is a known irritant). Approximately 0.02 ml of the test product was applied under an occlusive patch on the backof each subject. Patches remained in place for 24 h, and the sites were evaluated several minutes after patch removal. After evaluation, a new patch was applied to the same site. This procedure was continued for 21 consecutive days, excluding Sundays. The sites were scored on a scale of 0 (no reaction) to 4 (intense erythema with edema and vesicles). Of the 25 test subjects, two received scores of 1 (mild erythema), one on day 12 and one on day 15. The total cumulative score for the lip makeup containing 10% Arachidyl Propionate was 2, compared with 67 for 0.5% SLSand 519 for 4% SLS, and the lip makeup containing 10% Arachidyl Propionate was considered nonirritating to minimally irritating under the conditions of the study. The irritation and sensitization potential of a lip makeup containing 10% Arachidyl Propionate was evaluated using a modified Draize-Shelanski-Jordan repeat insult patch test.(201A total of 207 test subjects completed the study. The test sample (amount not specified) was applied under an occlusive patch to the back of each subject, patches remained in place for 24 h, and the sites were evaluated upon patch removal. Patches were applied every other day, Monday through Friday, for a total of ten induction applications. A 2 week nontreatment period followed the last induction application. After the nontreatment period a challenge patch was applied to a new site. This patch remained in place for 48 h, and the test areas were evaluated after patch removal. A second challenge patch was applied a week later, remaining in place for 48 h. The test sites were evaluated after patch removal and again 24 h later. Test sites were scored on a scale of O-4, with a score of 0 indicating no reaction, and a score of 4 indicating intense erythema with edema and vesicles. No reactions were observed in any of the test subjects during either the induction or challenge phases of the study, and the lip makeup containing 10% Arachidyl Propionate was considered neither an irritant nor a sensitizer under the conditions of the study.

Phototoxicity The phototoxicity of a lip makeup containing 10% Arachidyl Propionate was evaluated in 10 human subjects. (21)The lip makeup, 0.02 ml, was applied to duplicate sites on the back of each test subject. One site was covered, and the other site received 2-4 min [approximately 6 unfiltered minimal erythemal doses (MED)] of window glass-filtered UVA radiation from a Blue Point hot quartz spot lamp, followed by a 0.5 MED of UVB radiation. No details on the wavelength or irradiance of the light were given. Evaluations were then made of both the irradiated and nonirradiated sites, including a site which was irradiated but to which no lip makeup was applied. The sites were then covered and were reevaluated 1, 3, and 24 h after irradiation. Reactions were scored on a scale of O-4, with 0 indicating no reaction, and 4 indicating intense erythema with edema and vesicles. No adverse reactions were noted in any of the test subjects, and the lip makeup containing 10% Arachidyl Propionate was considered nonphototoxic.

Photoallergenicity A photoallergenicity test of a lip makeup containing 10% Arachidyl Propionatewas performed using 25 test subjects between the ages of 18 and 65.‘22’ The test substance,

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0.02 ml, was applied under an occlusive patch on either side of the spinal column. Patches remained in place for 24 h. Upon patch removal, the test site on one side of the spinal column was irradiated for 2-4 min with window-glass filtered light from a Blue Point solar simulating lamp. In addition, a nonpatched site was also irradiated. Evaluations of skin reactions were made a few minutes after exposure to the UV light. This procedure was repeated twice a week for 2 l/2 weeks, for a total of five applications. This was followed by a 12-day nontreatment period. At the end of the nontreatment period, patches were applied to new sites on the back, remaining in place for 24 h. After patch removal, one patch site, as well as one nonpatched site, were irradiated and the test sites were then evaluated for signs of irritation, and again evaluated 24 h later. Skin reactions were scored on a scale of O-4, with 0 indicating no reaction, and 4 indicating intense erythema with edema and vesicles. No reactions were noted in any of the test subjects at any time during the study. The lip makeup containing 10% Arachidyl Propionate was not considered a photoallergen under the conditions of the study.

SUMMARY

Arachidyl Propionate is an amber-colored semisolid wax. In ultraviolet spectral studies, it absorbed strongly in the 200-260 nm region, and absorbed little above 290 nm. Arachidyl Propionate is used in cosmetic formulations at concentrations of up to 10% to provide emolliency, gloss, and film-forming capabilities to the finished product. It has also been used as a substitute for lanolin and lanolin derivatives. According to the toxicity classification of Hodge and Sterner,‘231Arachidyl Propi- onate is practically nontoxic when ingested. No systemic toxicity was associated with subchronic oral exposure to Arachidyl Propionate. The substance was also nontoxic by dermal absorption. In test animals, Arachidyl Propionate was neither a primary dermal irritant, nor a skin sensitizer, nor did it have comedogenic potential. It was nonirritating to rabbit eyes. In clinical studies, Arachidyl Propionate was neither an irritant nor a sensitizer. In additional clinical studies, Arachidyl Propionate was neither phototoxic nor a photoallergen.

DISCUSSION

Arachidyl Propionate has some structural and chemical similarities to the stearates, for which a safety assessment has already been published. Although there is limited toxicological data available for Arachidyl Propionate, the data available in the stearates report are relevant to the safety evaluation of Arachidyl Propionate. Structure-activity relationships would indicate that Arachidyl Propionate would not absorb in the UVA and UVB ranges and spectral analyses as well as clinical studies support this conclusion.

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CONCLUSION

On the basis of the data included in this report and in the stearates report, the CIR Expert Panel concludes that Arachidyl Propionate is safe as a cosmetic ingredient in the present practices of use and concentration.

ACKNOWLEDGMENT

Julie K. Poudrier, Scientific Analyst and Writer, prepared the scientific literature review and technical analysis of this report.

REFERENCES

1. ELDER, R.L. (Editor). (1985). Final Report on the Safety Assessment of Butyl Stearate, Cetyl Stearate, lsobutyl Stearate, lsocetyl Stearate, Isopropyl Stearate, Myristyl Stearate, and Octyl Stearate. j. Am. Coil. Toxicol. 4(5):107-46. 2. WICKHEN PRODUCTS, INC. (1987). Arachidyl Propionate: Specifications Submission of data to CTFA.* 3. WICKHEN PRODUCTS, INC. (1987). Arachidyl Propionate: Material safety data sheet. Submission of data to CTFA.* 4. CUILLOT, J.P., MARTINI, M.C., and GIAUFFRET, J.Y. (1977). Safety evaluation of cosmetic raw materials. J. Sot. Cosmet. Chem. Z&377-93. 5. CASCHEM, INC. (1988). UV spectrum of Arachidyl Propionate. Submission of unpublished data to CTFA.* 6. CASCHEM, INC. (1989). UV spectrum of Arachidyl Propionate. Submission of unpublished data to CTFA.* 7. FOOD AND DRUG ADMINISTRATION (FDA). (1987). Cosmetic product formulation data. FDA computer printout. 8. CODE OF FEDERAL REGULATIONS. (1982). Title 21, Part 720.4. 9. WICKHEN PRODUCTS, INC. (1987). Letter to CTFA Industry Liaison accompanying submission of unpublished data.* 10. MB RESEARCH LABORATORIES, INC. (1975). Arachidyl Propionate: Oral LD,, in rats, primarydermal irritation, rabbit eye irritation, acute dermal toxicrty, and guinea pig sensitization. Project no. MB 75-776. Submission of unpublished data to CTFA.* 11. COSMETIC, TOILETRY, AND FRAGRANCE ASSOCIATION (CTFA). (1987). The safety evaluation of one raw ingredient: Arachidyl Propionate. 13 week subchronic oral toxicity study in albino rats. Study project code no. 252. Submission of unpublished data to CTFA.* 12. GUILLOT, J.P., CIAUFFRET, J.Y., MARTINI, M.C., CONNET, J.F., and SOULE, G. (1980). Safetyevaluation ofcosmetic raw materials: results obtained with 160 samples from various origm. IFREB Lancaster CED. Submission of data to CTFA.* 13. CTFA. (1985). Primary skin irritation, eye irritatton, and acute oral toxicity test results. Test Nos. S 15-108, E 58-062, and 36-115. Submission of unpublished data to CTFA.’ 14. MB RESEARCH LABORATORIES, INC. (1984). Comedogenicity assay in rabbits. Project no. MB 84-7290. Submission of unpublished data to CTFA.’ 15. CTFA. (198.5). Allergic contact sensitization test. Test no. 387-85. Submission of unpublished data to CTFA.* 16. CTFA. (1985). 4-Day Mini-Cum Results. Study ref. no. 391-85. Submission of unpublished data to CTFA.* 17. CTFA. (1985). Clinical test results of lipstick containing Arachidyl Propionate. Test no. 389-85. Submission of unpublished data to CTFA.* 18. UNIVERSITY OF CALIFORNIA, LOS ANGELES (UCLA). (1985). Supervised usage test of a lip makeup containing 10% Arachidyl Propionate. UCLA no. M-85.U.035. Submtssion of unpublished data to CTFA.* 19. UCLA. (19851. Twenty-one day cumulative irritation test of a lip makeup containing 10% Arachidyl Propionate. UCLA no. M-85-9l.U01-M-85-91.U22. Submission of unpublished data to CTFA.’ 20. UCLA.fl985). Repeatinsultpatchtestofalipmakeupcontaining lO%Arachidyl Propionate. UCLAno. M-85-19. Submission of unpublished data to CTFA.* 21. UCLA. (1985). Phototoxicity test of a lip makeup containing 10% Arachidyl Propionate. UCLA no. M-85-l 9T. Submission of unpublished data to CTFA.’ 22. UCLA. (1985). Photoallergenicity test of a lip makeup containing 10% Arachidyl Propionate. UCLA no. M-85-19P. Submission of unpublished data to CTFA.* 23. HODGE, H.C., and STERNER, J.H. (1949). Tabulation of toxicity classes. Am. Indus. Hyg. A. Quart. 10:934.

*Available for review: Director, Cosmetic Ingredient Review 1110 VermontAve., NW, Suite810, Washington, D.C. 20005.

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JOURNAL OF THE AMERICAN COLLEGE OF TOXICOLOGY Volume 11, Number 1,1992 Mary Ann Liebwt, Inc., Publishers

Final Report on the Safety Assessment of Isopropyl lsostearate

Isopropyl Isostearate, the ester of isopropyl alcohol and isostearic acid, is used as a skin conditioning agent-emollient in cosmetic products. Undiluted Isopropyl Iso- stearate was classified as a slight ocular irritant. Repeated applications of a 10.0% aqueous suspension of Isopropyl lsostearate to the skin of albino rabbits was well-tolerated. Based upon the safety test data on chemically similar isopropyl esters that is summarized in the report, it is concluded that Isopropyl lsostearate is safe as a cosmetic ingredient in the present practices of use and concentration.

INTRODUCTION

he toxicity of Isopropyl lsostearate is reviewed in this report. A summary of the T toxicity data on such chemically similar cosmetic ingredients as Isopropyl Stearate, Isopropyl Palmitate, Isopropyl Myristate, and Isopropyl Lanolate is also included as these have been reviewed previously by the CIR Expert Panel. The available safety data on these ingredients are considered to be supportive of the safety data on Isopropyl Isostearate.

CHEMISTRY

Chemical and Physical Properties

Isopropyl lsostearate (CAS Nos. 31478-84-9 and 68171-33-5), also known as Heptadecanoic Acid, 16-Methyl-, 1 -Methylethyl Ester, is the ester of isopropyl alcohol and isostearic acid that conforms generally to the formula:“’

0 II C,,H,C-OCH(CHJ,

It is a mixture of isopropyl esters of 80% branched chain C,, and C,, acids and 20% straight chain C,4, C,6, and C,, acids with a small amount of oleic acid.‘2’ Properties of Isopropyl lsostearate are summarized in Table 1.

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44 COSMETIC INGREDIENT REVIEW

TABLE 1. PROPERTIESOF ISOPROPYL ISOSTEARATE'*)

Form Liquid Solubility Soluble in acetone, ethyl acetate, isopropyl alcohol, and mineral oil Specific gravity at 25°C 0.853 to 0.859 Acid value 1 .O maximum Ester value 169.0 to 180.0 Iodine value 3.0 maximum

Analytical Methods

Isopropyl lsostearate has been analyzed via infrared spectroscopy and gas chromatography.‘2,3’

COSMETIC USE

Isopropyl lsostearate is used as a skin conditioning agent-emollient in cosmetics.‘4’ The FDA cosmetic product formulation computer printout”’ is compiled through voluntary filing of such data in accordance with Title 21 part 720.4 of the Code of Federal Regulations. w Ingredients are listed in preset concentration ranges under specific product type categories. Since certain cosmetic ingredients are supplied by the manufacturer at less than 100% concentration, the value reported by the cosmetic formulator may not necessarily reflect the actual concentration found in the finished product; the actual concentration would be a fraction of that reported to the FDA. Data submitted within the framework of preset concentration ranges provide the opportunity for overestimation of the actual concentration of an ingredient in a particular product. An entry at the lowest end of a concentration range is the same as one entered at the highest end of that range, thus introducing the possibility of a two- to ten-fold error in the assumed ingredient concentration. Isopropyl lsostearate is used in 78 cosmetic products at concentrations ranging from d 0.1% to 50.0% (Table 2).‘5’ Isopropyl lsostearate has been approved for use as a cosmetic ingredient in Japan.“’ This ingredient is not included in the list of substances that may not be used in cosmetic products marketed in countries of the European Economic Community.@’

TABLE 2. PRODUCT FORMULATION DATA FOR ISOPROPYL ISOSTEARATE"'

No. of product iormulations Total no of Total no. within each concentration range (%I formulations containing Product category in category ingredient >25-50 > 10-25 >5-10 >l-5 x.1-1 CO. J

Eye area cosmetics 1135 11 1 9 1 Facial makeup products 3194 40 1 7 8 19 4 1 Skin care products 3110 20 5 6 5 4 Miscellaneous other 2228 7 2 4 1 cosmetic products

1989 TOTALS 78 1 9 14 38 10 6

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ASSESSMENT: ISOPROPYL ISOSTEARATE 45

Noncosmetic Use

Isopropyl lsostearate is used as a dispersing agent in aerosol formulations.“’

TOXICOLOGY

Ocular Irritation

The ocular irritation potential of undiluted Isopropyl lsostearate was evaluated using six male New Zealand albino rabbits. The test substance was instilled (0.1 ml) into the conjunctival sac of one eye of each animal; eyes were not rinsed. Untreated eyes served as controls. Ocular irritation reactions were scored on days 1, 2, 3, 4, and 7 post-instillation according to the scale by Draize:“” O-l 10. A 10% aqueous suspension of Isopropyl lsostearate was also tested. Both concentrations of Isopropyl Isostear- ate were classified as slight ocular irritants.“”

Skin Irritation

The skin irritation potential of undiluted Isopropyl lsostearate was evaluated using six male New Zealand albino rabbits (2.5-3.5 kg). The test substance (0.5 ml) was applied via gauze pads to the left flank (intact) and right flank (abraded) of each animal. Each pad was covered with an occlusive patch and secured with hypoallergenic adhesive plaster. Sites were scored 24 and 72 h after patch application according to the scale: 0 (no edema, erythema) to 4 (severe edema, erythema). Isopropyl lsostearate was classified as a non-irritant (Primary Irritation Index = 0.42).‘“’ Isopropyl Isostearate, 0.5 ml of an undiluted and a 10.0% aqueous suspension, was applied daily to the right and left flanks (shaved skin) of each of three male albino rabbits (2.5-3.5 kg) fivedays per week for a period of 8 weeks. Excess test substance was wiped from the skin with gauze 30 s after each application. Application sites were scored daily according to the scale: 0 (no erythema) to 4 (serious erythema with slight scar formation) and 0 (no edema) to 4 (serious edema, covering all of the treated area, or more). Scores were averaged on a weekly basis, and the maximum irritation index (IIMM) was determined at the end of the treatment period. Microscopic examination was performed on skin samples (2 per animal) that differed macroscopically. The criteria for classifying a substance as poorly tolerated after repeated applications were as follows: (1) pathologic reactions must be observed in all three animals treated, (2) pathologic reactions must appear over the entire treated surface of the epidermis, and not only at localized points, and (3) microscopic examination of two fragments of treated skin from each rabbit must confirm the macroscopic observations. Undiluted Isopropyl Isostear- ate was very poorly tolerated (IIMM = 3.34; treatment wasdiscontinued after 5 weeks). Isopropyl lsostearate as a 10.0% aqueous suspension was relatively well tolerated (IIMM = 2.00).‘3’

Comedogenicity

The comedogenic potential of Isopropyl lsostearate (undiluted) was evaluated using two New Zealand albino rabbits. The test substance was applied to the internal surface of the external ear daily for 2 weeks. The untreated external ear served as the control. At the end of the treatment period, reactions were scored according to the scale: 0 (no

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46 COSMETIC INGREDIENT REVIEW

increase in visible follicular keratosis) to 5 (the presence of more severe cornedones throughout the ear). Isopropyl lsostearate induced the formation of severe cornedones throughout the skin of the external ear.‘12’

SUMMARY OF DATA ON CHEMICALLY SIMILAR INGREDIENTS

Acute Inhalation Toxicity

Neither death nor other signs of systemic toxicity was observed in rats exposed for 1 hour to an aerosol antiperspirant containing 16 to 20% isopropyl myristate. The gravimetric ambient concentration of isopropyl myristate at the time of exposure ranged from 33 to 41 mg/L of air. Similar results were reported in a study in which albino rats were exposed to aerosolized isopropyl palmitate (200 mg/L of air) for 1 h.

Acute Oral Toxicity

The LD,, was not achieved when albino rats were treated with 8 ml/kgof undiluted isopropyl stearate. When rats and mice were dosed with undiluted isopropyl myristate, the LD,, was not achieved at a dose of 16 ml/kg in rats and 49.7 ml/kg in mice. Additionally, doses of 64 ml/kg of undiluted isopropyl palmitate (rats) and 64 ml/kg of undiluted isopropyl lanolate (rats) were less than the LD,,.

Dermal Toxicity

A dermal LD,, of greater than 5 g/kg was reported in a study in which rabbits were treated with undiluted isopropyl myristate. No deaths were reported after rabbits received repeated applications of an aerosol antiperspirant containing 16 to 20% isopropyl myristate; severe skin irritation was observed. In another study, toxic effects were not observed in rabbits that received a single dermal application (2.0 ml/kg) of isopropyl palmitate, and no deaths were reported. Repeated applications of undiluted isopropyl palmitate also did not cause death. Orthokeratosis and slight acanthosis were cutaneous alterations observed in some of the rabbits tested. The repeated applications of a cosmetic foundation product containing 5% isopropyl lanolate did not induce signs of toxicity in rats.

Ocular Irritation

Undiluted isopropyl stearate did not induce ocular irritation when instilled into the conjunctival sac of New Zealand rabbits. Similar results were reported when undiluted isopropyl myristate was used. In five separate Draize ocular irritation tests, undiluted isopropyl palmitatedid not cause significant ocular irritation in any of the rabbits tested. In four separate ocular irritation studies using isopropyl lanolate, the mean Draize ocular irritation scores indicated no ocular irritation or mild transient irritation.

Skin Irritation

Moderate skin irritation was observed at intact and abraded skin sites of rabbits that received a single application of undiluted isopropyl stearate. In four Draize skin

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ASSESSMENT: ISOPROPYL ISOSTEARATE 47

irritation tests, repeated applications of undiluted isopropyl myristate to the skin of rabbits resulted in minimal irritation; results from a fifth Draize test indicated no skin irritation. Draize test results for isopropyl palmitate indicated that it was non-irritating to slightly irritating to the skins of albino rabbits. In other Draize tests, isopropyl lanolate was non-irritating to intact and abraded skin sites of rabbits and guinea pigs. A personal cleanliness product containing 1% isopropyl stearate was classified as a slight skin irritant after repeated applications were made to human subjects. Primary skin irritation was not observed in human subjects patch tested with undiluted isopropyl myristate. Repeated applications of this ingredient induced very slight skin irritation. In separate studies, primary skin irritation also was not observed in subjects patch tested with undiluted isopropyl palmitate or in those patch tested with undiluted isopropyl lanolate.

Skin Sensitization

Isopropyl myristate and isopropyl lanolate did not induce sensitization in guinea pigs when either was tested at a concentration of 0.1%. Skin sensitization was not observed in human subjects patch tested with two personal cleanliness products containing 1% isopropyl stearate. In separate maximization tests, skin sensitization was not observed in human subjects patch tested using 20% isopropyl myristate in petrolatum, those patch tested using a bath formulation containing 42.9% isopropyl myristate, or a bath formulation containing 45.6% isopropyl palmitate. In other studies, sensitization reactions were not observed in human subjects patch tested with undiluted isopropyl palmitate. The results from four sensitization studies of an aerosol antiperspirant containing 52 to 58% isopropyl myristate also were negative. Sensitization reactions were observed in two of 53 subjects challenged with undiluted isopropyl lanolate. It was noted that the two subjects were hypersensitive to many substances.

PHOTOSENSITIZATION

In a photosensitization study, distilled Isopropyl Lanolate was applied to the skin of 10 male guinea pigs daily for 10 days. After each application, the skin was exposed to ultraviolet light for 6 h. There was no evidence of photosensitization.

PHOTOTOXICITY

Phototoxicity was not observed in 10 healthy human subjects after patch testing (occlusive patches) with a bath formulation containing 42.9% isopropyl myristate in one study and after testing with 45.6% isopropyl palmitate in a second study. Test sites were irradiated after 6 h and 24 h of contact. Similar negative results were reported in photocontact allergenicity studies of these bath formulations.

CARCINOGENICITY

No neoplasms were induced after 0.1 ml of a 1% solution (solvent not stated) of isopropyl myristate was applied to the skins of mice once per week for 18 weeks. In

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48 COSMETIC INGREDIENT REVIEW

another study, isopropyl myristate was applied to the backs of mice twice per week from the time they were 7 weeks old until death (average lifespan = approximately 80 weeks). Isopropyl myristate concentrations of 10, 50, and 100% in acetone were applied to groups of 50 animals.

SUMMARY

Isopropyl lsostearate (CAS Nos. 31478-84-9 and 68171-33-5) is the ester of isopropyl alcohol and isostearic acid. It is used as askin conditioning agent-emollient in cosmetics at concentrations ranging from d 0.1% to 50.0%. Undiluted Isopropyl lsostearate and a 10.0% aqueous suspension of Isopropyl lsostearate were classified as slight ocular irritants when instilled into the conjunctival sac of albino rabbits. A single 24 h application, under an occlusive patch, of undiluted Isopropyl lsostearate did not cause skin irritation in albino rabbits. Repeated applications of undiluted Isopropyl lsostearate and a 10.0% aqueous suspension of Isopropyl Isostear- ate to the skin of albino rabbits were very poorly tolerated and relatively well tolerated, respectively. Cornedones were observed in the skin of the external ear of albino rabbits that received repeated applications of undiluted Isopropyl Isostearate.

DISCUSSION

Data indicating that undiluted Isopropyl lsostearate is a skin irritant and a slight ocular irritant, and that 10.0% Isopropyl lsostearate is a slight ocular irritant, but not a skin irritant, are presented in this report. These data, along with positive comedogenicity data, are the only toxicological data available for the CIR Expert Panel’s review of Isopropyl Isostearate. The Expert Panel hasdetermined that chemically similar cosmetic ingredients, isopropyl stearate, isopropyl myristate, isopropyl palmitate, and isopropyl lanolate, are safe, at concentrations used in cosmetics.“3-‘6’ These ingredients, including Isopropyl Isostearate, are isopropyl esters resulting from the reaction of isopropyl alcohol with long-chain saturated aliphatic acids. Like Isopropyl Isostearate, cosmetic use concentrations of these ingredients range from d 0.1% to 50.0%. A collective evaluation of the data on animals summarized in this report indicates that the isopropyl esters, undiluted or at use concentrations, are relatively harmless when administered to animals in acute oral, dermal, and inhalation toxicity studies, are not significant ocular irritants, induce skin irritation reactions ranging from none to moderate, are not sensitizers or photosensitizers, and are not carcinogenic. In studies involving healthy human subjects, these esters, undiluted or at use concentrations, produced, at most, slight skin irritation but not sensitization. Based upon chemical similarities between the isopropyl esters and similar cosmetic use concentrations, the Expert Panel determined that the data used in its safety assessments of isopropyl stearate, isopropyl myristate, isopropyl palmitate, and isopropyl lanolate can also be used in the safety assessment of Isopropyl Isostearate.

CONCLUSION

On the basis of the available animal and clinical data on isopropyl stearate, isopropyl myristate, isopropyl palmitate, and isopropyl lanolate presented in this

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ASSESSMENT: ISOPROPYL ISOSTEARATE 49

report, the CIR Expert Panel concludes that Isopropyl lsostearate is safe as a cosmetic ingredient in the present practices of use and concentration.

ACKNOWLEDGEMENT

This report was prepared by Wilbur Johnson, Jr., Senior Scientific Analyst and Writer.

REFERENCES

1. ESTRIN, N.F., CROSLEY, P.A., and HAYNES, C.R. (1982). Cosmetic Ingredient Dictmnary. Washington DC: The Cosmetic Toiletry and Fragrance Association. Inc., p. 138. 2. ESTRIN, N.F., HAYNES, CR., and WHELAN, J.M. (1982). Cosmetic Ingredient Specifications. CTFA Compendium of

Cosmetic Ingredient Composition. WashIngton. DC: The Cosmetic, Toiletry and Fragrance Association, Inc. 3. GUILLOT, J.P., GIAUFFRET, J.Y., MARTINI, M.C., CONNET, J.F.. and SOULE, C. (1981). Raw materials used in

cosmetology. Informations Chimie 213, 239-249. 4. NIKITAKIS, J.M. (Editor). (1988). CJFA Cosmetic/ngred/entHandbook. First Edition. Washington, DC: TheCosmetic, Toiletry

and Fragrance Association, Inc. p. 237. 5. FOOD AND DRUG ADMINISTRATION (FDA). (1989). Cosmetic product formulation data on isopropyl isostearate. FDA

computer prmtout. 6. CODE OF FEDERAL REGULATIONS (CFR). (Revised as of April 1, 1984). Title 21 Part 720.4. Voluntary filing of cosmettc

product ingredient and cosmetic raw material cornpositIon statement. Information requested about cosmetic products.

Washington, DC: Government Printing Office. 7. NIKKOL CHEMICALS CO., LTD. (1989). Cross Reference Index: An Index Cross-Referencing the English Names of Cosmetic

lngredtents in Japan with CTFA Adopted Names. Nikko1 Chemicals Co., Ltd. Tokyo, Japan.

8. DUPUIS, J. (1989). The European Economic Community (EEC) Cosmetics Directive. Updated Version-incorporating all

amendments up to March 15, 1989.

9. SCIARRA, J.J., IANNACONE, A., and MORES, L. (1976). An evaluation of dispersing agents in aerosol formulations. I. Synthetic esters. J. Sot. Cosmet. Chem. 27, 209-20. 10. DRAIZE, J.H. (1959). Scaleforscoringocular lesions. In: Appraisal of!heSafetyoiChemicafs in Foods, Drugs, andCosmetics.

Austin, TX: The Association of Food and Drug Officials of the United States, p. 51.

11. GUILLOT, J.P.. CIAUFFRET, J.Y., MARTINI, M.C., CONNET, J.F., and SOULE, G. (1980). Safety evaluation ofcosmetic raw materials: results obtained with 160 samples from various origin. pp. 109, 110, 105-108, and 123.

12. FULTON, I.E., JR., PAY, S.R., and FULTON, I.E., Ill. (1984). Comedogenicity ofcurrenttherapeutic products, cosmetics, and ingredients in the rabbit ear. J. Am. Acad. Dermatol. 10(l), 96-105. 13. ELDER, R.L. (Editor). (1985). Final report on the safety assessment of butyl stearate, cetyl stearate, isobutyl stearate, isocetyl

stearate, isopropyl stearate, myristyl stearate, and octyl stearate. J. Am. Coil. Toxicol. 4(5), 107-146. 14. ELDER, R.L. (Editor). (1982). Final report on the safety assessment of myrlstyl myristate and isopropyl myristate. J. Am. Co/l.

Toxicol. l(4), 55-80. 15. ELDER, R.L. (Editor). (1982). Final report on the safety assessment of octyl palmitate, cetyl palmitate, and isopropyl palmitate.

1. Am. Coil. Toxicol. l(2), 13-35.

16. ELDER. R.L. (Editor). (1980). Final report of the safety assessment for isopropyl lanolate. J. Environ. Pathol. Toxicol. 4(4): 121-132.

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International Journal of Toxicology, 26(Suppl. 3):31–77, 2007 Copyright c American College of Toxicology ISSN: 1091-5818 print / 1092-874X online DOI: 10.1080/10915810701663150 Final Report on the Safety Assessment of Ricinus Communis (Castor) Seed Oil, Hydrogenated Castor Oil, Glyceryl Ricinoleate, Glyceryl Ricinoleate SE, Ricinoleic Acid, Potassium Ricinoleate, Sodium Ricinoleate, Zinc Ricinoleate, Cetyl Ricinoleate, Ethyl Ricinoleate, Glycol Ricinoleate, Isopropyl Ricinoleate, Methyl Ricinoleate, and Octyldodecyl Ricinoleate1

skin, but not irritating to miniature swine skin. Ricinoleic Acid was The oil derived from the seed of the Ricinus communis plant nonirritating in mice and in one rabbit study, but produced well- and its primary constituent, Ricinoleic Acid, along with certain of defined erythema at abraded and intact skin sites in another rabbit its salts and esters function primarily as skin-conditioning agents, study. Zinc Ricinoleate was not a sensitizer in guinea pigs. Neither emulsion stabilizers, and surfactants in cosmetics, although other castor oil nor Sodium Ricinoleate was genotoxic in bacterial or functions are described. Ricinus Communis (Castor) Seed Oil is mammalian test systems. Ricinoleic Acid produced no neoplasms the naming convention for castor oil used in cosmetics. It is pro- or hyperplasia in one mouse study and was not a tumor promoter in duced by cold pressing the seeds and subsequent clarification of another mouse study, but did produce epidermal hyperplasia. Cas- the oil by heat. Castor oil does not contain ricin because ricin does tor oil extract had a strong suppressive effect on S180 body tumors not partition into the oil. Castor oil and Glyceryl Ricinoleate ab- and ARS ascites cancer in male Kunming mice. No dose-related sorb ultraviolet (UV) light, with a maximum absorbance at 270 nm. reproductive toxicity was found in mice fed up to 10% castor oil Castor oil and Hydrogenated Castor Oil reportedly were used in for13weeks. Female rats injected intramuscularly with castor oil 769 and 202 cosmetic products, respectively, in 2002; fewer uses on the first day after estrus had suppressed ovarian folliculogenesis were reported for the other ingredients in this group. The highest and anti-implantation and abortive effects. Castor oil used as a ve- reported use concentration (81%) for castor oil is associated with hicle control in rats receiving subcutaneous injections had no effect lipstick. Castor oil is classified by Food and Drug Administration on spermatogenesis. A methanol extract of Ricinus communis var. (FDA) as generally recognized as safe and effective for use as a minor seeds (ether-soluble fraction) produced anti-implantation, stimulant laxative. The Joint Food and Agriculture Organization anticonceptive, and estrogenic activity in rats and mice. Clinically, (FAO)/World Health Organization (WHO) Expert Committee on castor oil has been used to stimulate labor. Castor oil is not a signif- Food Additives established an acceptable daily castor oil intake icant skin irritant, sensitizer, or photosensitizer in human clinical (for man) of 0 to 0.7 mg/kg body weight. Castor oil is hydrolyzed in tests, but patients with occupational dermatoses may have a pos- the small intestine by pancreatic enzymes, leading to the release of itive reaction to castor oil or Ricinoleic Acid. The instillation of a glycerol and Ricinoleic Acid, although 3,6-epoxyoctanedioic acid, castor oil solution into the eyes of nine patients resulted in mild 3,6-epoxydecanedioic acid, and 3,6-epoxydodecanedioic acid also and transient discomfort and minor epithelial changes. In another appear to be metabolites. Castor oil and Ricinoleic Acid can en- study involving 100 patients, the instillation of castor oil produced hance the transdermal penetration of other chemicals. Although corneal epithelial cell death and continuity breaks in the epithelium. Because castor oil contains Ricinoleic Acid as the primary fatty acid chemically similar to prostaglandin E1, Ricinoleic Acid did not have the same physiological properties. These ingredients are not acute group, the Cosmetic Ingredient Review (CIR) Expert Panel consid- toxicants, and a National Toxicology Program (NTP) subchronic ered the safety test data on the oil broadly applicable to this entire oral toxicity study using castor oil at concentrations up to 10% in group of cosmetic ingredients. The available data demonstrate few the diet of rats was not toxic. Other subchronic studies of castor toxic effects. Although animal studies indicate no significant irri- oil produced similar findings. Undiluted castor oil produced min- tant or sensitization potential, positive reactions to Ricinoleic Acid imal ocular toxicity in one study, but none in another. Undiluted in selected populations with identified dermatoses did suggest that castor oil was severely irritating to rabbit skin in one study, only sensitization reactions may be higher in that population. Overall, slightly irritating in another, mildly irritating to guinea pig and rat however, the clinical experience suggests that sensitization reactions are seen infrequently. In the absence of inhalation toxicity data on these ingredients, the Panel determined that these ingredients can be used safely in aerosolized cosmetic products because Received 14 May 2007; accepted 24 August 2007. the particle sizes produced are not respirable. Overall, the CIR Address correspondence to Wilbur Johnson, Jr., Senior Scientific Expert Panel concluded that these cosmetic ingredients are safe in Analyst and Writer, Cosmetic Ingredient Review, 1101 17th Street, the practices of use and concentrations as described in this safety NW, Suite 412, Washington, DC 20036, USA. assessment. 1Reviewed by the Cosmetic Ingredient Review Expert Panel.

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INTRODUCTION fatty acyl groups (not specified) (TNO BIBRA International Ltd. The safety of the following ingredients in cosmetics is re- 1999). viewed in this report: Ricinus Communis (Castor) Seed Oil, According to other sources, Castor Oil contains 2.4% lau- Hydrogenated Castor Oil, Glyceryl Ricinoleate, Glyceryl Ri- ric acid (Larsen et al. 2001), 2% to 5% linoleic acid (Maier cinoleate SE, Ricinoleic Acid, Potassium Ricinoleate, Sodium et al. 1999), and globulin, cholesterol, lipase, vitamin E, and Ricinoleate, Zinc Ricinoleate, Cetyl Ricinoleate, Ethyl Rici- β-sitosterol (Scarpa and Guerci 1982). noleate, Glycol Ricinoleate, Isopropyl Ricinoleate, Methyl Ri- cinoleate, and Octyldodecyl Ricinoleate. For the sake of brevity, Hydrogenated Castor Oil Ricinus Communis (Castor) Seed Oil will be referred to as castor According to the International Cosmetic Ingredient Dictio- oil. nary and Handbook (Gottschalck and McEwen 2004), Hydro- Glyceryl Ricinoleate and Glyceryl Ricinoleate SE are es- genated Castor Oil (CAS no. 8001-78-3) is defined as the end ters of glycerin and Ricinoleic Acid (component of castor oil). product of controlled hydrogenation of Ricinus Communis (Cas- A Cosmetic Ingredient Review (CIR) safety assessment (Elder tor) Seed Oil. Castor Oil, Hydrogenated is another technical 1988) of Glyceryl Ricinoleate in cosmetics was published with name for this ingredient, and Castor Wax (trade name) and Mon- the conclusion that the available data are insufficient to support tane 481 (trade name mixture) are other names under which Hy- the safety of this ingredient as used in cosmetics. The follow- drogenated Castor Oil is marketed (Gottschalck and McEwen ing data were needed for completion of this safety assessment: 2004). (1) 28-day chronic dermal toxicity (guinea pigs) and (2) clinical sensitization and photosensitization studies (or an appropriate Ricinoleic Acid, Its Salts, and Simple Esters ultraviolet spectrum instead of the photosensitization data). Table 1 includes the structures, definitions, and cosmetic Glyceryl Ricinoleate and Glyceryl Ricinoleate SE are in- ingredient functions of Glyceryl Ricinoleate, Glyceryl Rici- cluded in the present safety assessment in the expectation that noleate SE, Ricinoleic Acid, Potassium Ricinoleate, Sodium the available data on castor oil, Ricinoleic Acid, salts of Rici- Ricinoleate, Zinc Ricinoleate, Cetyl Ricinoleate, Ethyl Rici- noleic Acid, and other esters of Ricinoleic Acid will contribute noleate, Glycol Ricinoleate, Isopropyl Ricinoleate, Methyl Ri- to a resolution of the CIR Expert Panel’s data needs for Glyceryl cinoleate, and Octyldodecyl Ricinoleate. Ricinoleate and Glyceryl Ricinoleate SE. Chemical and Physical Properties CHEMISTRY Properties of castor oil and Hydrogenated Castor Oil are summarized in Table 2 and properties of the following ingre- Definition and Structure dients are summarized in Table 3: Ricinoleic Acid, Ethyl Ri- Ricinus Communis (Castor) Seed Oil cinoleate, Methyl Ricinoleate, Sodium Ricinoleate, Potassium According to the International Cosmetic Ingredient Dictio- Ricinoleate, and Zinc Ricinoleate. nary and Handbook (Gottschalck and McEwen 2004), Ricinus Communis (Castor) Seed Oil (CAS no. 8001-79-4) is defined as the fixed oil that is obtained from the seeds of Ricinus communis. Composition/Impurities Other names for this oil include: Castor Oil, Castor Oil (Ricinus Castor Oil communis L.), Castor Seed Oil, Ricinus Communis, and Ricinus The following three peaks were observed in a gas-liquid chro- Communis Oil (Gottschalck and McEwen 2004), and Ricinus matogram on castor oil: palmitic acid (1.7%), C18 acids (stearic, Oil (TNO BIBRA International Ltd. 1999). As given by TNO oleic, and linoleic acids) (15.9%), and Ricinoleic Acid (82.4%) BIBRA International Ltd. (1999), the structural formula is: (Kato and Yamaura 1970). According to the Food Chemicals Codex (National Academy CH2OR of Sciences 1996), the requirements for castor oil are as follows: || free fatty acids (passes test), heavy metals, as Pb (not more than CHOR 10 mg/kg), hydroxyl value (between 160 and 168), iodine value || (between 83 and 88), saponification value (between 176 and CH2OR 185), and specific gravity (between 0.952 and 0.966). The test for free fatty acids is described as follows: free fatty acids (10 g) are where R represents a fatty acyl group [CH3(CH2)5CH(OH) dissolved in 50 ml of a mixture of equal volumes of alcohol and CH2CH=CH(CH2)7COOH] that is typically derived from Ri- ether (which has been neutralized to phenolphthalein with not cinoleic Acid. Ricinoleic Acid accounts for 87% to 90% of more than 7 ml of 0.1 N sodium hydroxide). Phenolphthalein (1 the fatty acyl groups, and the following other fatty acids com- ml) is added, followed by titration with 0.1 N sodium hydroxide prise the remaining fatty acyl groups: oleic acid (2% to 7%), until the solution remains pink (after shaking). linoleic acid (3% to 5%), palmitic acid (1% to 2%), stearic Similarly, the United States Pharmacopeia (Committee of acid (1%), dihydrostearic acid (1%), and trace amounts of other Revision of the United States Pharmacopeial Convention 2004a)

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TABLE 1 Structures, deﬁnitions, and functions of ricinoleic acid, its salts, and esters (Gottschalck and McEwen 2004) Structure Deﬁnition Function

CH3(CH2)5CHCH2CH CH(CH2)7COOH Ricinoleic Acid (CAS nos. 141-22-0 and 7431-95-0) is an Surfactant–cleansing

OH unsaturated fatty acid. Other names include: agent 12-Hydroxy-9-Octadecenoic Acid; 9-Octadecenoic Acid, 12-Hydroxy-; Ricinic Acid; and Ricinolic Acid OH Glyceryl Ricinoleate (CAS nos.141-08-2, 1323-38-2, and Skin-conditioning 5086-52-2) is the monoester of glycerin and ricinoleic acid. agent—emollient and CH(CH2)5CH3 O Other names include: Glycerin Monoricinoleate; Glycerol surfactant– CH CH CH(CH ) C OCH CHCH OH 2 2 7 2 2 Monoricinoleate; Glyceryl Monoricinoleate, emulsifying agent OH 12-Hydroxy-9-Octadecenoic Acid, Monoester with 1,2,3-Propanediol; Ricinoleic Acid Monoglyceride; and Ricinolein, 1-Mono- See above Glyceryl Ricinoleate SE is a self-emulsifying grade of Glyceryl Skin-conditioning Ricinoleate containing sodium and/or potassium stearate agent—emollient and surfactant– emulsifying agent CH3(CH2)5CHCH2CH CH(CH2)7COOK Potassium Ricinoleate (CAS no. 7492-30-0) is the potassium salt Surfactant–cleansing OH of Ricinoleic Acid. Other names include: agent and surfactant– 12-Hydroxy-9-Octadecenoic Acid, Monopotassium Salt and emulsifying 9-Octadecenoic Acid, 12-Hydroxy-, Monopotassium Salt agent See above, except Sodium Ricinoleate (CAS no. 5323-95-5) is the sodium salt of Surfactant–cleansing with Na in place Ricinoleic Acid. Other names include: agent and surfactant– of K 12-Hydroxy-9-Octadecenoic Acid, Sodium Salt; 9-Octadecenoic emulsifying agent Acid, 12-Hydroxy, Sodium Salt; and Sodium Ricinate Zinc Ricinoleate (CAS no. 13040-19-2) is the zinc salt of Anticaking agent, - CH3(CH2 )5CHCH2CH CH(CH2)7COO Ricinoleic Acid. Other names include: deodorant agent, and Zn+2 OH 12-Hydroxy-9-Octadecenoic Acid, Zinc Salt and opacifying agent

2 9-Octadecenoic Acid, 12-Hydroxy-, Zinc Salt OH Cetyl Ricinoleate (CAS no.10401-55-5) is the ester of cetyl Skin-conditioning

CH3(CH2) 5CHCH2CH O alcohol and Ricinoleic Acid. Other names include: Hexadecyl agent—occlusive

CH(CH2)7C OCH2(CH2)14CH3 12-Hydroxy-9-Octadecenoate; 12-Hydroxy-9-Octadecenoic Acid, Hexadecyl Ester; 9-Octadecenoic Acid, 12-Hydroxy-, Hexadecyl Ester; and Ricinoleic Acid, Hexadecyl Ester O Ethyl Ricinoleate (CAS no. 55066-53-0) is the ester of ethyl Fragrance ingredient CH3(CH2)5CHCH2CH CH(CH2)7C OCH2CH3 alcohol and Ricinoleic Acid. Other names include: Ethyl and skin-conditioning OH 12-Hydroxy-9-Octadecenoate; 12-Hydroxy-9-Octadecenoic agent—emollient O Acid-, Ethyl Ester; 9-Octadecenoic Acid, 12-Hydroxy-, Ethyl CH (CH ) CHCH CH CH(CH ) C 3 2 5 2 2 7 OCH2CH3 Ester; and Ricinoleic Acid, Monoethyl Ester OH O

CH3(CH2)5CHCH2CH CH(CH2)7C OCH2CH3

OH OH Glycol Ricinoleate (CAS nos. 106-17-2 and 40275-40-9) is the Emulsion stabilizer CH(CH2 )5 CH3 O ester of ethylene glycol and Ricinoleic Acid. Other names and skin-conditioning

CH2 CH CH(CH2 )7 COCH2 CH2 OH include: 1,2-Ethanediol Monoricinoleate; Ethylene Glycol agent—emollient Monoricinoleate; Glycol Monoricinoleate; 2-Hydroxyethyl 12-Hydroxy-9-Octadecenoate; 9-Octadecenoic Acid, 12-Hydroxy-, 2-Hydroxyethyl Ester; and Ricinoleic Acid, 2-Hydroxyethyl Ester (Continued on next page)

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TABLE 1 Structures, deﬁnitions, and functions of ricinoleic acid, its salts, and esters (Gottschalck and McEwen 2004) (Continued) Structure Deﬁnition Function

O Isopropyl Ricinoleate (CAS no. 71685-99-9) is the ester of Skin-conditioning CH (CH ) CHCH CH 3 2 5 2 CH(CH2)7C OCHCH3 isopropyl alcohol and Ricinoleic Acid. Other names include: agent—emollient

OH CH3 12-Hydroxy-9-Octadecenoic Acid, 1-Methylethyl Ester; 1-Methylethyl, 12-Hydroxy-9-Octadecenoate; 9-Octadecenoic Acid, 12-Hydroxy-, 1-Methylethyl Ester; and Ricinoleic Acid, Isopropyl Ester O Methyl Ricinoleate (CAS nos. 141-24-2 and 23224-20-6) is the Skin-conditioning

CH3(CH2)5CHCH2CH CH(CH2)7COCH3 ester of methyl alcohol and ricinoleic acid. Other names include: agent—emollient

OH Castor Oil Acid, Methyl Ester; 12-Hydroxy-9-Octadecenoic Acid, Methyl Ester; Methyl Castor Oil Fatty Acid; Methyl 12-Hydroxy-9-Octadecenoate; 9-Octadecenoic Acid, 12-Hydroxy-, Methyl Ester; and Ricinoleic Acid, Methyl Ester CH3(CH2)5CHCH2CH O Octyldodecyl Ricinoleate (CAS nos. 79490-62-3 and 125093-27-8) Skin-conditioning OH CH(CH2)7COCH2CH(CH2)9CH3 is the ester of Octyldodecanol and Ricinoleic Acid. Other names agent—occlusive

(CH2)7CH3 include: 12-Hydroxy-9-Octadecenoic Acid, 2-Octyldodecyl Ester; 9-Octadecenoic Acid, 12-Hydroxy-, 2-Octyldodecyl Ester; 2-Octyldodecyl 12-Hydroxy-9-Octadecenoate; and 2-Octyldodecyl Ricinoleate has published the following requirements for castor oil: free fatty This residue, known as pomace, contains ricin and a separate acids (“the free fatty acids in 10 g require for neutralization not allergen (Ratner and Gruehl 1929). Following extraction from more than 3.5 ml of 0.10 N sodium hydroxide”) heavy metals the castor bean, neither of these two fractions is present in castor (0.001%), hydroxyl value (between 160 and 168), iodine value oil (Ordman 1955). (between 83 and 88), saponification value (between 176 and According to a more recent reference (Cornell University 182), and specific gravity (between 0.957 and 0.961). 2001), ricin does not partition into the oil because of its water solubility. Therefore, castor oil does not contain ricin, provided Hydrogenated Castor Oil that cross-contamination does not occur during its production. According to Nikitakis and McEwen (1990), Hydrogenated Other sources indicate that castor oil is produced via the cold Castor Oil consists primarily of glyceryl-tri-hydroxystearate. pressing of the seeds of Ricinus communis (Hui 1996) and by An earlier source (Binder et al. 1970) indicates that cold expression and subsequent clarification of the oil by heat 12-hydroxystearic acid is the principal constituent of Hydro- (Gennaro 1990). genated Castor Oil. The National Formulary (Committee of Revision of the Hydrogenated Castor Oil United States Pharmacopeial Convention 2004b) has published the following requirements for Hydrogenated Castor Oil: free According to Nikitakis and McEwen (1990), Hydrogenated fatty acids (free fatty acids in 20 g require for neutralization not Castor Oil is obtained by the controlled hydrogenation of pure more than 11.0 ml of 0.1 N sodium hydroxide), heavy metals Castor Oil. Another source (Campbell & Co. 2002) indicates (0.001%), hydroxyl value (between 154 and 162), iodine value that Hydrogenated Castor Oil (hard, brittle wax) results from the (not more than 5), and saponification value (between 176 and addition of hydrogen to Castor Oil in the presence of a nickel 182). catalyst.

Methods of Production Ricinoleic Acid Castor Oil One of the simplest methods for obtaining Ricinoleic Acid is According to Kathren et al. (1959), castor oil is extracted the hydrolysis of castor oil. Ricinoleic acid may then be sepa- from the bean of the tropical plant, Ricinus communis,byeither rated from hydrolyzed castor Oil by successive additions of urea of the following two methods: (1) use of a solvent [not stated] (i.e., urea complexing) (Chakravarty and Bose 1963). Accord- or (2) mechanical crushing, grinding, and pressing. The former ing to a more recent source, Ricinoleic Acid results from the method is more efﬁcient, leaving a more dessicated residue. saponiﬁcation of castor oil (Lewis 1997).

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TABLE 2 Chemical and physical properties of Castor Oil and Hydrogenated Castor Oil Property Value Reference Castor Oil Color/Form Colorless to pale-yellow viscous liquid Lewis 2000 Taste Slightly acrid National Toxicology Program (NTP) 2003 Specific gravity 0.945 to 0.965 at 25◦/25◦C; 0.961 to 0.963 at NTP 2003 15.5◦/15.5◦C Density 0.953 to 0.965 g/ml at 20◦C NTP 2003 Viscosity 6 to 8 poises at 25◦C NTP 2003 283 cP at 37◦C Fredholt et al. 2000 Solubility In water (<1 mg/ml at 20◦C); in DMSO (≥100 NTP 2003 mg/ml at 20◦C); In 95% ethanol (≥ 100 mg/ml at 20◦C); in methanol (miscible); In acetone (≥100 mg/ml at 20◦C) Miscible in absolute alcohol, glacial acetic acid, Lewis 2000 chloroform, and ether Surface tension 39.0 dynes/cm at 20◦C; 35.2 dynes/cm at 80◦C NTP 2003 Refractive index 1.4784 at 20◦C; 1.473 to 1.477 at 25◦C; NTP 2003 1.466 to 1.473 at 40◦C Optical rotation Not less than +3.5◦ NTP 2003 Flash point 229◦C (445◦F) NTP 2003 Autoignition 448◦C (840◦F) NTP 2003 temperature Melting point −12◦C NTP 2003 Boiling point 313◦C NTP 2003 Freezing point −10◦C NTP 2003 Saponification 178 NTP 2003 value Iodine value 85 NTP 2003 Acid value <4 NTP 2003 Reichert-Meissl <0.5 NTP 2003 value Polenske value <0.5 NTP 2003 Acetyl value 144 to 150 NTP 2003 Hydroxyl value 161 to 169 NTP 2003 Hydrogenated Castor Oil Color/Form White to cream waxy solid Nikitakis and McEwen 1990 Odor As specified by the buyer Nikitakis and McEwen 1990 Slight characteristic odor Allegri et al. 1981 Taste Tasteless Allegri et al. 1981 Identification Positive: Close match to a standard IR spectrum Nikitakis and McEwen 1990 with no indication of foreign materials Specific gravity 0.98 to 1.04 at 25◦/25◦C Nikitakis and McEwen 1990 Melting range 78◦ to 90◦C Nikitakis and McEwen 1990 Acid value 5.0 maximum Nikitakis and McEwen 1990 Saponification 175 to 185 Nikitakis and McEwen 1990 value Iodine value As specified by the buyer Nikitakis and McEwen 1990 <5.0 Budavari 1989 Hydroxyl value 155 max KIC Chemicals, Inc. 2004 (Continued on next page)

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TABLE 2 Chemical and physical properties of Castor Oil and Hydrogenated Castor Oil (Continued) Property Value Reference Density 0.98 g/cm3 Physical & Theoretical Chemistry Lab 2004 Solubility Insoluble in water and organic solvents. Lewis 1997 Soluble in chloroform; very slightly soluble in ben- Allegri et al. 1981 zene; virtually insoluble in alcohol, ethyl ether, carbon disulﬁde, and water Drying loss When dried under vacuum in presence of phosphoric Allegri et al. 1981 acid anhydride, should lose no more than 0.1% of weight Sulfuric ash Not more than 0.01% Allegri et al. 1981

Methyl Ricinoleate According to Gunstone (1989), Castor Oil is a pre- According to Masri et al. (1962), Methyl Ricinoleate has cursor of octadeca-9,11-dienoic acid (dehydration reaction); been obtained by alcoholysis of castor oil, followed by fractional 12-hydroxystearic acid (hydrogenation); heptanol and undec- distillation of the crude esters and further purification by low 10-enoic acid (pyrolysis); and sebacic acid, 10-hydroxydecanoic temperature crystallization. According to a more recent source, acid, octan-2-ol, and octan-2-one (alkaline fusion). Methyl Ricinoleate is a product of either the esterification of According to Achaya (1971), the hydrogenation of Castor Ricinoleic Acid or the alcoholysis of castor oil; the product is oil produces 12-hydroxystearate, stearate, 12-ketostearate, and purified by vacuum distillation (Lewis 1997). ricinoleyl alcohol. Dehydrated castor oil is a dehydration product of castor oil, and contains a fair proportion of conjugated dienes. Ultraviolet Absorption Castor Oil Hydrogenated Castor Oil A ultraviolet (UV) absorption spectrum on castor oil indicates Hydrogenated Castor Oil is combustible and is incompatible maximum absorbance at 270 nm; absorption peaks at 280 and with strong oxidizing agents (Physical and Theoretical Chem- 260 nm were also observed (Sasol North America, Inc. no date). istry Lab 2004).

Glyceryl Ricinoleate USE AUVabsorption spectrum on Glyceryl Ricinoleate indicates maximum absorbance at 270 nm and an absorption peak Purpose in Cosmetics at 280 nm (Sasol North America, Inc. no date). Ricinus Communis (Castor) Seed Oil functions as a fragrance ingredient and a skin-conditioning agent—occlusive in Analytical Methods cosmetic products, and Hydrogenated Castor Oil functions as a Castor Oil skin-conditioning agent—occlusive and a viscosity increasing Castor oil has been analyzed using the following meth- agent—nonaqueous (Gottschalck and McEwen 2004). The cos- ods: mass spectroscopy (Ayorinde et al. 2000), gas-liquid metic ingredient functions of Ricinoleic Acid and some of its chromatography (Kato and Yamaura 1970; Ramsey et al. salts and esters are included in Table 1. Except for the anticaking, 1980), thin-layer chromatography (Srinivasulu and Mahapatra deodorant, and opacifying agent functions of Zinc Ricinoleate 1973), and nonaqueous reverse-phase high-performance liquid and the surfactant function of Ricinoleic Acid, Potassium Rici- chromatography–mass spectrometry (St¨ubigeret al. 2003). noleate, Sodium Ricinoleate, Glyceryl Ricinoleate, and Glyceryl Ricinoleate SE, these ingredients function as skin-conditioning Methyl Ricinoleate agents in cosmetic products. In addition to functioning as a Methyl Ricinoleate has been analyzed by nuclear magnetic skin-conditioning agent, Ethyl Ricinoleate also functions as a resonance (NMR) spectroscopy (Wineburg and Swern 1973) and fragrance ingredient. Glycol Ricinoleate also functions as an gas-liquid chromatography (Paulus and Champion 1972). emulsion stabilizer, and Glyceryl Ricinoleate and Glyceryl Ri- cinoleate SE also function as surfactants in cosmetics. Reactivity Castor Oil Extent of Use in Cosmetics Castor oil is combustible when exposed to heat, and sponta- Frequency of use data based on industry reports to the Food neous heating may occur (Lewis 2000). and Drug Administration (FDA) in 2002 indicate that Ricinus

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TABLE 3 Chemical and physical properties of Ricinoleic Acid, its salts, and esters Property Value Reference Ricinoleic Acid Molecular weight 298.47 Lide and Frederikse 1993 Color/Form Yellow fatty acid Grant 1972 Yellow, viscous liquid Nikitakis and McEwen 1990 Colorless to yellow, viscous liquid Lewis 1997 Odor As specified by the buyer Nikitakis and McEwen 1990 Identification Positive: Close match to a standard IR spectrum with no in- Nikitakis and McEwen 1990 dication of foreign materials Specific gravity 0.940 at 25◦/25◦C Nikitakis and McEwen 1990 Density 0.9450 Lide and Frederikse 1993 0.940 at 27.4◦/4◦CLewis 1997 Solubility Insoluble in water Grant 1972 Soluble in alcohol, acetone, ether, and chloroform Budavari 1989 Soluble in ethyl alcohol and diethyl ether Lide and Frederikse 1993 Soluble in most organic solvents; insoluble in water Lewis 1997 Refractive index 1.4716 at 20◦C Nikitakis and McEwen 1990 1.4716 Lide and Frederikse 1993 1.4697 at 20◦CLewis 1997 Optical rotation [α]22/D of +6.67◦C Budavari 1989 +6.67◦ at 22◦C Nikitakis and McEwen 1990 [α]12/Dof +5.05 Lide and Frederikse 1993 Dextrorotatory Lewis 1997 Melting point 5.5◦C Nikitakis and McEwen 1990 α :7.7◦C; β:16◦C; γ : 5.5◦C Lide and Frederikse 1993 5.5◦CLewis 1997 Boiling point 226.8◦C Lide and Frederikse 1993 226◦Cat10mmHgLewis 1997 Combustibility Combustible Lewis 1997 Neutralization value 187.98 Budavari 1989 188 Nikitakis and McEwen 1990 Iodine value 85.05 Budavari 1989 85 Nikitakis and McEwen 1990 Potassium Ricinoleate Color/Form White paste Lewis 1997 Solubility Soluble in water Lewis 1997 Combustibility Combustible Lewis 1997 Sodium Ricinoleate Color/Form White or slightly yellow powder Lewis 1997 Odor Nearly odorless Lewis 1997 Solubility Soluble in water or alcohol Lewis 1997 Combustibility Combustible Lewis 1997 Zinc Ricinoleate Color/Form Fine white powder Lewis 1997 Odor Faint fatty acid odor Lewis 1997 Density 1.10 at 25◦/25◦CLewis 1997 (Continued on next page)

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TABLE 3 Chemical and physical properties of Ricinoleic Acid, its salts, and esters (Continued) Property Value Reference Melting point 92◦Cto95◦CLewis 1997 Combustibility Combustible Lewis 1997 Methyl Ricinoleate Color/Form Colorless liquid Lewis 1997 Solubility Insoluble in water; soluble in alcohol and ether Lewis 1997 Density 0.9236 at 22◦/4◦CLewis 1997 Refractive index 1.4628 Lewis 1997 Boiling point 245◦Cat10mmHgLewis 1997 Flash point −4.5◦CLewis 1997 Combustibility Combustible Lewis 1997 Ethyl Ricinoleate Molecular weight 326.52 Lide and Frederikse 1993 Density 0.9045 Lide and Frederikse 1993 Refractive index 1.4618 Lide and Frederikse 1993 Optical rotation [α]22/D of +5.3 Lide and Frederikse 1993 Boiling point 258◦C Lide and Frederikse 1993

Communis (Castor) Seed Oil and Hydrogenated Castor Oil are Cosmetic products containing Ricinus Communis (Castor) being used in a total of 769 and 202 cosmetic products, respec- Seed Oil, Hydrogenated Castor Oil, and Ricinoleic Acid and its tively. Glyceryl Ricinoleate (16 cosmetic products), Ricinoleic salts and esters are applied to most areas of the body, and could Acid (6 cosmetic products), Sodium Ricinoleate (12 cosmetic come in contact with the oral, ocular, or nasal mucosae. These products), Zinc Ricinoleate (3 cosmetic products), and Cetyl Ri- products may be used on a daily basis, and could be applied cinoleate (55 cosmetic products) were also reported. These data frequently over a period of several years. are given in Table 4 (FDA 2002). Product categories for Ricinus Communis (Castor) Seed Oil, Use concentration data obtained from an industry survey by Hydrogenated Castor Oil, and salts and esters of Ricinoleic Acid the Cosmetic, Toiletry, and Fragrance Association (CTFA 2004) include potential aerosol applications. Bower (1999) character- indicate that Ricinus Communis (Castor) Seed Oil and Hydro- ized the typical diameter of anhydrous hair spray particles in the genated Castor Oil are being used in cosmetics at concentrations 60- to 80-µm range (typically, <1% are below 10 µm). Pump up to 81% and 39%, respectively (see Table 4). hair sprays, in contrast, have typical particle diameters of ≥80 Other maximum ingredient use concentrations reported in µm. Johnsen (2004) reported that the mean particle diameter is Table 4 are as follows: Glyceryl Ricinoleate (up to 12%), Zinc around 38 µminatypical aerosol spray. In practice, he stated Ricinoleate (up to 2%), Cetyl Ricinoleate (up to 10%), and that aerosols should have at least 99% of particle diameters in Octyldodecyl Ricinoleate (up to 5%). Of the product categories the 10- to 110-µm range. For comparison, Jensen and O’Brien listed in Table 4, the highest reported use concentration for Rici- (1993) reported a mean aerodynamic diameter of 4.25 ± 1.5 µm nus Communis (Castor) Seed Oil is associated with lipsticks. for respirable particles, which is smaller than any of the cosmetic Two other sources indicate that lipstick contains 44% w/w product particle sizes given above. castor oil (Hui 1996) and 10% to 67% castor oil (Smolinske 1992). Certain ingredients in this group are reportedly used in a given Noncosmetic Use product category, but the concentration of use is not available. Castor Oil, Hydrogenated Castor Oil, and Ricinoleic Acid Salts For other ingredients in this group, information regarding use and Esters concentration for speciﬁc product categories is provided, but the According to Aplin and Eiseo (1997), castor oil is used as number of such products is not known. In still other cases, an an industrial lubricant and as a medicinal purgative; the au- ingredient is not in current use, but may be used in the future. thors noted that medicinal Castor Oil does not contain ricin. Mc- Forexample, Potassium Ricinoleate, Ethyl Ricinoleate, Glycol Keon et al. (2000) conﬁrmed lubricant and anti-fungal uses and Ricinoleate, Isopropyl Ricinoleate, and Methyl Ricinoleate are additional uses in paints, coatings, and plastics. not currently reported to FDA as in use, nor are there industry The following uses have been reported for Hydrogenated Cas- survey data indicating a current use concentration. tor Oil (Budavari 1989): in water-repellent coatings, candles,

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TABLE 4 Current uses and concentrations of Ricinus Communis (Castor) Seed Oil and Ricinoleic Acid and its salts and esters in cosmetics Ingredient uses in each Use concentrations Product category (total no. of formulations) product category (FDA 2002) (CTFA 2004) (%) Ricinus Communis (Castor) Seed Oil Baby products Shampoos (29) — — Lotions, oils, powders, and creams (60) 1 — Other (34) 2 — Bath products Oils, tablets, and salts (143) 1 22 Soaps and detergents (421) — 0.008 Bubble baths (215) — — Capsules (2) — — Other (196) 10 — Eye makeup Eyebrow pencils (102) 10 3 Eyeliners (548) 11 4–8 Eye shadow (576) 9 2–5 Eye lotions (25) — — Eye makeup remover (100) 1 — Mascara (195) 16 0.4–3 Other (152) 11 13 Fragrance products Colognes and toilet waters (684) — 0.1 Perfumes (235) — 0.3–1 Powders (273) — — Sachets (28) — — Other (173) — — Noncoloring hair care products Conditioners (651) 20 0.0002–5 Sprays/aerosol ﬁxatives (275) 1 — Straighteners (63) — — Permanent waves (207) — — Rinses (42) 5 19 Shampoos (884) — 0.005 Tonics, dressings, etc. (598) 34 0.005–52 Wave sets (53) — 16 Other (277) 6 — Hair—coloring products Dyes and colors (1690) 27 2 Tints (49) — — Rinses (20) — 0.0005 Shampoos (32) — — Color sprays (5) — — Lighteners with color (5) — — Bleaches (120) 1 0.04 Other (55) — — Makeup Blushers (245) 10 0.08–45 Face powders (305) 1 1 (Continued on next page)

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40 COSMETIC INGREDIENT REVIEW

TABLE 4 Current uses and concentrations of Ricinus Communis (Castor) Seed Oil and Ricinoleic Acid and its salts and esters in cosmetics (Continued) Ingredient uses in each Use concentrations Product category (total no. of formulations) product category (FDA 2002) (CTFA 2004) (%) Foundations (324) 5 0.05–6 Leg and body paints (4) — — Lipsticks (962) 492 15–81 Makeup bases (141) 4 0.002–20 Rouges (28) 1 38–48 Makeup ﬁxatives (20) — — Other (201) 25 40–49 Nail care products Basecoats and undercoats (44) — — Cuticle softeners (19) 1 — Creams and lotions (15) 1 — Nail extenders (1) — — Nail polishes and enamels (123) — 69 Nail polish and enamel removers (36) 8 — Other (55) 3 — Personal hygiene products Bath soaps and detergents (421) 18 ≤ 0.008 Underarm deodorants (247) — 0.01–3 Douches (5) — — Feminine deodorants (4) — — Other (308) 1 0.003 Shaving products Aftershave lotions (231) 2 0.004–0.03 Beard softeners (0) — — Mens talcum (7) — — Preshave lotions (14) — — Shaving creams (134) — — Shaving soaps (2) — — Other (63) 1 0.03 Skin care products Skin cleansing creams, lotions, liquids, and pads (775) 4 0.004 Depilatories (34) 2 — Face and neck creams, lotions, powders, and sprays (310) 1 1–7 Body and hand creams, lotions, powders, and sprays (840) 3 0.02–55 Body and hand sprays — 0.004 Foot powders and sprays (35) — — Moisturizers (905) 8 7 Night creams, lotions, powders, and sprays (200) — 0.00004 Paste masks/mud packs (271) 2 — Skin fresheners (184) — — Other (725) 8 — Suntan products Suntan gels, creams, liquids, and sprays (131) 1 6 Indoor tanning preparations (71) — — Other (38) 1 10 Total uses/ranges for Ricinus Communis (Castor) Seed Oil 769 0.00004–81

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TABLE 4 Current uses and concentrations of Ricinus Communis (Castor) Seed Oil and Ricinoleic Acid and its salts and esters in cosmetics (Continued) Ingredient uses in each Use concentrations Product category (total no. of formulations) product category (FDA 2002) (CTFA 2004) (%) Hydrogenated Castor Oil Baby products Other (34) — 5 Bath products Oils, tablets, and salts (143) 1 19 Soaps and detergents (421) 1 0.0003–0.7 Bubble baths (215) — 4 Other (196) — 3 Eye makeup Eyebrow pencils (102) 11 4–26 Eyeliners (548) 25 3–39 Eye shadow (576) 6 2–19 Eye lotions (25) — 0.5 Eye makeup Eye makeup remover (100) 1 7 Mascara (195) — 5 Other (152) 14 22–39 Fragrance products Colognes and toilet waters (684) — 0.0003 Perfumes (235) — 5 Other (173) — 7 Noncoloring hair care products Shampoos (884) — 0.02 Tonics, dressings, etc. (598) — 3 Other (277) 1 — Makeup Blushers (245) 2 3 Foundations (324) 4 0.9–5 Leg and body paints (4) — 1 Lipsticks (962) 20 0.5–33 Makeup ﬁxatives (20) — 3–7 Other (201) 24 0.04–13 Nail care products Creams and lotions (15) 1 — Oral hygiene products Other (6) — 1–5 Personal hygiene products Underarm deodorants (247) 2 2–8 Feminine deodorants (4) — 2 Other (308) 65 — Shaving products Aftershave lotions (231) 2 — Skin care products Skin cleansing creams, lotions, liquids, and pads (775) 3 1 Face and neck creams, lotions, powders, and sprays (310) — 1–2 Body and hand creams, lotions, powders, and sprays (840) 3 0.0003–1 Foot powders and sprays (35) — 8 (Continued on next page)

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42 COSMETIC INGREDIENT REVIEW

TABLE 4 Current uses and concentrations of Ricinus Communis (Castor) Seed Oil and Ricinoleic Acid and its salts and esters in cosmetics (Continued) Ingredient uses in each Use concentrations Product category (total no. of formulations) product category (FDA 2002) (CTFA 2004) (%) Moisturizers (905) 3 1–5 Night creams, lotions, powders, and sprays (200) 1 1–3 Other (725) 9 0.1–1 Suntan products Suntan gels, creams, liquids, and sprays (131) 1 0.8–3 Other (38) 2 — Total uses/ranges for Hydrogenated Castor Oil 202 0.0003–39 Glyceryl Ricinoleate Eyebrow pencil (102) — 11 Eyeliner (548) 1 2 Eyeshadow (576) 5 — Other eye makeup products (152) 1 2–12 Lipsticks (962) 5 — Other shaving products (63) 1 — Body and hand creams, lotions, powders, and sprays (840) 2 — Night creams, lotions, powders, and sprays (200) 1 — Total uses/ranges for Glyceryl Ricinoleate 16 12 Ricinoleic Acid Bath preparations Oils, tablets and salts (143) 1 — Fragrance preparations Colognes and toilet waters (684) 1 — Other fragrance preparations (173) 4 — Total uses/ranges for Ricinoleic Acid 6 — Sodium Ricinoleate Baby products Other baby products (34) 1 — Bath Preparations Soaps and detergents (421) 10 — Noncoloring Hair Preparations Shampoos (884) 1 — Total uses/ranges for Sodium Ricinoleate 12 — Zinc Ricinoleate Noncoloring hair preparations Hair sprays/aerosol ﬁxatives (275) — 1 Personal hygiene products Underarm deodorants (247) 2 2 Skin care preparations Skin cleansing creams, lotions, liquids, and pads (775) 1 — Body and hand skin care preparations (sprays) (840) — 1 Total uses/ranges for Zinc Ricinoleate 3 1–2 Cetyl Ricinoleate Makeup preparations Blushers (245) — 3

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CASTOR OIL 43

shoe polish, carbon paper, and ointments; for impregnating pa- FDA-approved direct/indirect food additive uses of Castor per, wood, and cloth; for electrical condenser impregnation; as oil and the following Ricinoleic Acid salts and esters are sum- a solid lubricant; and as a pressure mold release agent in the marized in Table 5: Potassium Ricinoleate, Sodium Ricinoleate, manufacture of formed plastics and rubber goods. The use of Zinc Ricinoleate, Glycol Ricinoleate, Isopropyl Ricinoleate, and Hydrogenated Castor Oil in waterproofing fabrics has also been Methyl Ricinoleate. reported (Lewis 1997). The Joint Food and Agriculture Organization (FAO)/World According to FDA’s OTC (Over-the-Counter) Drug Review Health Organization (WHO) Expert Committee on Food Addi- Ingredient Status Report (FDA 2003a), castor oil is classified tives (1980) established an acceptable daily intake (for man) of as generally recognized as safe and effective for use as a stimu- 0to0.7 mg/kg body weight for castor oil. The determination lant laxative, but not generally recognized as safe and effective of range was based on consideration of the lack of adequate for use as a wart remover. For use as a stimulant laxative, the long-term studies. single daily dose reported for one product ranges from 15 to 60 ml for adults and children ≥12 years (Drugstore.com, Inc. Ricinoleic Acid 2004). Noncosmetic uses of Ricinoleic Acid are as follows: turkey FDA had issued a proposed rule in 1982 stating that Castor oil red oil, textile finishing, source of sebacic acid and heptanol, is generally recognized as safe and effective and not misbranded ricinoleate salts, and 12-hydroxystearic acid (Lewis 1997). when used as an active ingredient (laxative) in OTC laxative drug products (FDA 1985) and a final rule is pending (FDA 2003a). Zinc Ricinoleate FDA has concluded that castor oil does not meet monograph The following noncosmetic uses of Zinc Ricinoleate have conditions and is not generally recognized as safe and effective been reported: fungicide, emulsifier, greases, lubricants, water- for use as a wart remover in OTC wart remover drug products proofing, lubricating-oil additive, and stabilizer in vinyl com- (FDA 1990). pounds (Lewis 1997). Castor oil is included in the list of inactive ingredients (excipients) present in approved oral, intramuscular, and topical Methyl Ricinoleate drug products or conditionally approved drug products that are Noncosmetic uses of Methyl Ricinoleate include: plasticizer, currently marketed for human use (FDA 2003b). lubricant, cutting oil additive, and wetting agent (Lewis 1997).

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TABLE 5 Direct/indirect food additive uses of Castor Oil, Hydrogenated Castor Oil, and Ricinoleic acid salts and esters Code of Federal Use Specification/listing Regulations (CFR) Diluent Not more than 500 ppm Castor Oil in the finished food may 21 CFR 73.1 be used safely as a diluent in color additive mixtures for food use that are exempt from certification. Release/antisticking agent Provided that Castor Oil meets the specifications of the 21 CFR 172.876 United States Pharmacopeia, it is permitted for use as a release agent and antisticking agent, not to exceed 500 ppm in hard candy. Natural flavoring substance Castor Oil is included in the list of natural flavoring 21 CFR 172.510 and adjuvant substances and natural adjuvants that are permitted for direct addition to food for human consumption. Plasticizer Castor Oil is approved for use as a plasticizer (total not to 21 CFR 177.2600 exceed 30% by weight of rubber product, unless otherwise specified) in rubber articles that have been declared safe for use in producing, manufacturing, packing, processing, preparing, treating, packaging, transporting, or holding food. Component of food-contact Hydrogenated Castor Oil may be used safely as a 21CFR176.170 surface component of the uncoated or coated food-contact surface of paper and paperboard intended for use in producing, manufacturing, packaging, processing, preparing, treating, packing, transporting, or holding aqueous and fatty foods. Component of food-contact Hydrogenated Castor Oil may be used safely as a 21CFR176.180 surface component of the uncoated or coated food-contact surface of paper and paperboard intended for use in producing, manufacturing, packaging, processing, preparing, treating, packing, transporting, or holding dry food. Component of defoaming Castor Oil, Hydrogenated Castor Oil, Glycol Ricinoleate, 21 CFR 176.210 agents Isopropyl Ricinoleate, and Methyl Ricinoleate are among the substances that are permitted for use in the formulation of defoaming agents that have been declared safe for use in the manufacture of paper and paperboard intended for use in packaging, transporting, or holding food. Component of surface Castor Oil is included in the list of substances permitted for 21 CFR 178.3910 lubricants use in surface lubricants that have been declared safe for use in the manufacture of metallic articles that contact food. May be used in surface lubricants employed to facilitate the drawing, stamping, or forming of metallic articles from rolled foil or sheet stock by further processing, provided that the total residual lubricant remaining in the metallic article in the form in which it contacts food does not exceed 0.2 mg per square inch of food-contact surface. Component of cellophane Hydrogenated Castor Oil is included in the list of 21CFR 177.1200 components of cellophane that may be safely used for packaging food.

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TABLE 5 Direct/indirect food additive uses of Castor Oil, Hydrogenated Castor Oil, and Ricinoleic acid salts and esters (Continued) Code of Federal Use Specification/listing Regulations (CFR) Component of Hydrogenated Castor Oil may be employed in the 21CFR177.1210 closure-sealing gaskets manufacture of closure-sealing gaskets that may be safely used on containers intended for use in producing, manufacturing, packing, processing, preparing, treating, packaging, transporting, or holding food. Component of cross-linked Hydrogenated Castor Oil may be used in the production of 21CFR 177.2420 polyester resins or may be added to cross-linked polyester resins that may be safely used as articles or components of articles that are intended for repeated use in contact with food. Textiles and textile fiber Fats, oils, fatty acids, and fatty alcohols derived from Castor 21 CFR 177.2800 components derived from Oil are included in the list of substances that are permitted Castor Oil for use in textiles and textile fibers that have been declared safe for use as articles or components of articles that are intended for use in producing, manufacturing, packing, processing, preparing, treating, packaging, transporting, or holding food. Component of adhesives Polymeric esters of polyhydric alcohols and polycarboxylic 21 CFR 175.105 acids prepared from glycerin and phthalic anhydride and modified with Castor Oil are permitted for use in adhesives that have been declared safe for use as components of articles intended for use in packaging, transporting, or holding food. Methyl Ricinoleate is also permitted for use in these adhesives. Component of resinous and Castor Oil, Hydrogenated Castor Oil, Potassium 21 CFR 175.300 polymeric coatings Ricinoleate, Sodium Ricinoleate, and Zinc Ricinoleate are included in the list of substances employed in the production of resinous and polymeric coatings that have been declared safe for use as the food-contact surface of articles intended for use in producing, manufacturing, packing, processing, preparing, treating, packaging, transporting, or holding food.

BIOLOGICAL PROPERTIES obtained from the liver, small intestine, and muscle; glyceride fatty acids were obtained from the liver and fat depots. There Absorption, Distribution, Metabolism, and Excretion wasnoevidence of catharsis in any of the animals. Castor Oil Average percentages of Ricinoleic Acid in the phospholipid In a study by Paul and McKay (1942), two rabbits (weight fatty acids were as follows: liver (test: 1.3 ± 0.6% [9 analy- = 3 kg) were fed 6% castor oil in the diet for 18 days; fecal ses]; controls: 1.7 ± 1.1% [7 analyses]), small intestine (test: collection occurred during the last ten days. The utilization (un- 4.9 ± 1.7% [8 analyses]; controls: 6.0 ± 4.4% [4 analyses]), corrected for metabolic fat) of castor oil was 92.1%, which the and skeletal muscle (test: 3.6 ± 2.9% [8 analyses]; controls: authors considered to be efﬁcient utilization. For both rabbits, 4.0 ± 1.7% [7 analyses]). The following values are average the percentage of fat in the feces was 2.2%. percentages of Ricinoleic Acid in glycerides and cholesterol es- In a study by Stewart and Sinclair (1945), adult rats (number, ters: fat depots (test: 6.8 ± 4.2% [11 analyses]; controls: 0.5 ± weights, and strain not stated) received a diet containing 48.4% 0.5% [7 analyses]) and liver (test: 7.2 ± 2.4% [8 analyses]; con- castor oil for 4 to 6 weeks. Control rats received stock ration trols: 5.6 ± 4.1% [5 analyses]). only. Feces were collected from three rats on the castor oil diet. The authors concluded that the feeding of castor oil did not At the end of the feeding period, excised organs/tissues were lead to the appearance of signiﬁcant amounts of Ricinoleic Acid ground thoroughly and samples of phospholipid fatty acids were in phospholipids of the small intestine, liver, and skeletal muscle,

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46 COSMETIC INGREDIENT REVIEW nor in glycerides of the liver. Additionally, they concluded that centage of Ricinoleic Acid in the feces, i.e., greater absorption ricinoleic acid is a component acid of the glycerides in the fat at the lower dose (Watson and Gordon 1962). depots, comprising 7% of the total fatty acids. The fatty acids Castor oil is metabolized to Ricinoleic Acid by pancreatic excreted by each of three rats amounted to 2.1%, 2.2%, and lipase in hamsters (Gaginella and Bass 1978). Thompson (1980) 3.6% of those ingested. Total body fat in these three animals reported that Castor Oil is a triglyceride that is hydrolyzed in was also determined, and it was calculated that 1% to 2% of the small intestine in humans by pancreatic enzymes, leading to absorbed Ricinoleic Acid was deposited in the fat depots. The the release of glycerol and Ricinoleic Acid. authors concluded that Ricinoleic Acid was rapidly metabolized In a study by Hagenfeldt et al. (1986), castor oil was ad- (Stewart and Sinclair 1945). ministered intragastrically to germ-free and conventional rats Watson and Gordon (1962) studied the digestion, absorption, (number not stated). Urine was collected at intervals over a and metabolism of castor oil (medicinal grade) in eight male 24-h period. The following epoxydicarboxylic acids were de- Sprague-Dawley rats (weights = 100 to 200 g). The composition tected in the urine of both germ-free and conventional rats: of the castor oil was as follows: ricinoleic acid (90.0%), linoleic 3,6-epoxyoctanedioic acid; 3,6-epoxydecanedioic acid; and 3,6- acid (4.7%), oleic acid (3.2%), stearic acid (1.0%), palmitic acid epoxydodecanedioic acid. These acids were not detected in urine (1.0%), and palmitoleic acid (0.1%). In the first experiment, four collected from the rats prior to dosing with castor oil, and they rats were fed rat chow ad libitum, and the remaining four were also were not detected in steam-sterilized castor oil. The authors fasted overnight. On the following morning, castor oil (1.0 ml) stated that results for the germ-free rat indicate that the cycliza- was dosed via stomach tube and chyle was collected over a 24- tion of Ricinoleic Acid (hydroxy fatty acid in castor oil) to form h period. The mean values for percent recovery of Ricinoleic an epoxy compound occurs endogenously and does not require Acid in fasted and fed rats were 6.8% and 24.2%, respectively the presence of intestinal bacteria. (p <.01). In a study by Ihara-Watanabe et al. (1999), two groups of five In the second experiment, seven weanling rats were fed a diet male Wistar rats (3 weeks old) received 10% castor oil in the consisting of rat chow that had been mixed with castor oil (20% diet (cholesterol-enriched and cholesterol-free, respectively) for by weight). The control group was fed an olive oil-supplemented 20 days. In both dietary groups, a very small quantity of Rici- diet. After 4 and 8 weeks of feeding, an epididymal fat pad was noleic Acid was present in perirenal adipose tissue, but not in removed from each rat in both groups, and fatty acid composition the serum or hepatic tissue. It was also noted that the perirenal was determined using gas-liquid chromatography. Mean values fatty acid profiles did not reflect those of the dietary fats, either for Ricinoleic Acid in the fat pad after 4 and 8 weeks of feeding in the absence or presence of dietary cholesterol. The fecal re- were 9.1 ± 1.7 and 9.7 ± 1.0%, respectively. Ricinoleic Acid was covery of Ricinoleic Acid was approximately 0.5% of the total undetectable in the fat pads of control rats fed olive oil. For rats ingested. It was concluded that castor oil was readily absorbed fed castor oil, random analyses of feces indicated a considerable and metabolized. fraction of hydroxystearic acid. However, hydroxystearic acid was undetectable in the feces of rats fed a normal diet. The Hydrogenated Castor Oil authors suggested that hydrogenation of Ricinoleic Acid in the In a study by Binder et al. (1970), groups of 15 male albino gut lumen by intestinal bacteria would be a likely explanation rats (Slonaker substrain of Wistar strain, weights = 43 to 83 g) for this finding. received diets containing the following oils: group 1 (fed 1% The relationship between dose and absorption of castor oil Hydrogenated Castor Oil (HCO) + 19% corn oil for 16 weeks), was studied in the final experiment. Polyethylene cannulae were group 2 (1% HCO + 19% corn oil for 8 weeks, then 20% corn oil inserted into the thoracic duct and duodenum of each of two for 8 weeks), group 3 (10% HCO + 10% corn oil for 16 weeks), rats, and the animals received 0.5 N saline overnight. On the and group 4 (10% HCO + 10% corn oil for 8 weeks, then 20% following morning, one rat received 0.2 ml and one rat received corn oil for 8 weeks). The control group received 20% corn oil in 0.6 ml of castor oil, and the rats were killed 45 min post dos- the diet for 16 weeks. The sample of HCO that was incorporated ing. Only the high dose induced diarrhea. Intestines (small and into the diet contained 86.5% 12-hydroxystearic acid, 10.3% large) were excised, homogenized, and tissue lipids were ex- nonoxygenated acids, and 3.2% 12-ketostearic acid. Thus, the tracted. Chyle was also collected and extracted. Total lipid esti- effective dietary concentrations of Hydrogenated Castor Oil in mation and gas-liquid chromatography (GLC) analysis of fatty the 1% and 10% diets were 0.865% and 8.65%, respectively. acids were performed on the extracts. For the rat dosed with 0.2 Results relating to the toxicity of HCO are summarized in the ml castor oil, the percentage of Ricinoleic Acid in the chyle was section Subchronic Oral Toxicity later in the report. 18.1% and the percentage detected in the small bowel lipids was After 4 weeks of feeding, some of the animals on the 1% HCO 6.2%. For the rat dosed with 0.6 ml Castor Oil, the percentage (three rats) and 10% HCO (three rats) diets were necropsied and of Ricinoleic Acid in the chyle was 3.0% and the percentage in excised abdominal adipose tissue from each group was pooled. the small bowel was 3.1%. Ricinoleic Acid was undetectable in At weeks 4 and 8, sets of three rats on the corn oil diet were used. large bowel lipids from the rat with diarrhea. The result indicates At the end of the 16-week feeding period, adipose tissue from a close correlation between the dose administered and the per- rats in the various dietary groups was analyzed. Adipose tissue

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CASTOR OIL 47 was not pooled. After 8, 12, and 16 weeks, lipids were extracted cumulation of hydroxy acids by 5% of fatty acids in fat tissue from three rats on each diet. Methyl esters were recovered from was noted. Analyses of adipose tissue indicated an occurrence the extracted lipids and chromatographed. of shorter chain hydroxy acids other than Ricinoleic Acid. The deposition of hydroxystearic acid in abdominal fat Ricinoleic Acid was also injected intraperitoneally to deter- and other body lipids was reported. In all cases, it was mine whether another route of administration would lead to the accompanied by hydroxypalmitic acid, hydroxymyristic acid, formation of shorter chain hydroxy acids. After Ricinoleic Acid and hydroxylauric acid (hydroxy-stearic acid metabolites). was injected intraperitoneally daily for 7 days, Ricinoleic Acid The percent composition of these HCO-derived hydroxy fatty accumulated in the adipose tissue; no shorter chain hydroxy acids in rat lipids was as follows: 12-hydroxystearic acid acids were reported (Okui et al. 1964). (≈81%), 10-hydroxypalmitic acid (≈17%), 8-hydroxymyristic In a study by Rao et al. (1969), Ricinoleic Acid or Methyl acid (≈1.6%), and 6-hydroxylauric acid (≈0.4%). After 4 weeks Ricinoleate was administered via gastric intubation to male rats on the 1% HCO diet, HCO-derived fatty acids accounted for (minimum weight of 400 g) with a cannulated thoracic duct. 0.90% (by weight) of the abdominal fat fatty acids. This propor- Lymph was collected for 48 h, and the lipids were then ex- tion decreased to ≈0.35% over the 8- to 16-week feeding period. tracted and separated into various lipid classes. Results indicated Compared to the abdominal fatty acids, the acids obtained from that Ricinoleic Acid was present in the triglyceride, diglyceride, the carcass lipids contained a smaller proportion of HCO-derived monoglyceride, and free fatty acid fractions. Peak absorption of hydroxyacids (≈0.28% over the 8- to 16-week feeding period). Ricinoleic Acid occurrred within 30 min post administration. Ri- The period encompassing 4 weeks of feeding on the 10% HCO cinoleic Acid was not present in the phospholipid or cholesterol diet yielded the greatest content of HCO-derived hydroxy acids ester fractions of the lymph lipids. in the lipids (i.e., 4.4% hydroxy acids in abdominal fat). At 16 weeks, this proportion had decreased to <2% (about same as in the carcass lipids). Percutaneous Absorption A rapid decrease in the amount of HCO-derived hydroxy fatty Ricinoleic Acid acids in the tissues was noted when the HCO diet was changed Butcher (1952) evaluated the skin penetration of Ricinoleic to the control diet (Binder et al. 1970). Acid in vivo using rats that were 20 to 30 days old. In order to increase the fluorescence of Ricinoleic Acid, either one part of methyl anthranilate or methylcholanthrene was added to 99 parts Ricinoleic Acid and Methyl Ricinoleate Ricinoleic Acid. The test substance was either gently rubbed Uchiyama et al. (1963) studied the accumulation of hydroxy on to skin that had been clipped free of hair. Biopsies were acids in depot fat after rats were fed with Ricinoleic Acid in taken at various intervals post application. The preparations were two experiments. In the first experiment, adult male rats (num- observed using a Spencer microscope with a quartz condenser. ber, strain, and weights not stated) were fed Ricinoleic Acid Ricinoleic Acid was retained mainly in the outer strata of the (5% emulsion, 20 ml) for 7 days. In the second experiment, the epidermis. There was little evidence of deeper penetration in animals were fed for 27 days. Lipid extraction from the fat tis- biopsies that were taken at2hpost application. sue was followed by hydrolysis to yield a fatty acid mixture. A In a study by Baynes and Riviere (2004), the percutaneous ab- gas-liquid chromatogram indicated appreciable amounts of the sorption of a radiolabeled [3H]Ricinoleic Acid (specific activity following hydroxy fatty acids with shorter chain lengths than = 20.0 mCi/mmol) mixture was evaluated using porcine skin Ricinoleic Acid: 10-hydroxyhexadecenoic acid (experiment 1: membranes or silastic (polydimethylsioloxane) membranes in 0.60% of total fatty acids; experiment 2: 0.33% of total fatty the Bronaugh flow-through diffusion cell system. [3H]Ricinoleic acids), 8-hydroxytetradecenoic acid (experiment 1: 0.03% of Acid (5%) mixtures were prepared in water containing either total fatty acids; experiment 2: 0.08% of total fatty acids), and 5% mineral oil or 5% PEG 200. Other [3H]Ricinoleic Acid 6-hydroxydodecenoic acid (experiment 2: 0.03% of total fatty mixtures were formulated with the following three commonly acids). Ricinoleic Acid comprised 0.51% of total fatty acids in used cutting fluid additives: triazine, linear alkylbenzene sul- experiment 1 and 3.85% of total fatty acids in experiment 2. fonate, and triethanolamine. At 8 h after topical exposure, Ri- In a study by Okui et al. (1964), the metabolism of hy- cinoleic Acid absorption (based on amount recovered in re- droxy fatty acids was evaluated using male albino rats (weight ceptor fluid) ranged from 1% to 13% in silastic membranes = 120 ± 10 g). The rats received 1.5 g Ricinoleic Acid or an and 0.1% to 0.3% in porcine skin membranes. For most mix- emulsion containing 5% (w/v) Ricinoleic Acid by stomach tube tures, peak absorption of Ricinoleic Acid occurred within 3 three times per day for a desired period. Feces were collected h. The greatest Ricinoleic Acid peak concentrations were as- every 24 h until the animals were killed. The animals were killed sociated with the control mixtures containing PEG in both 20 h after the last dose, and subcutaneous adipose tissue was re- membranes. moved for analyses. Oral dosing with Ricinoleic Acid resulted At the conclusion of the 8-h perfusion experiments, as much in a fecal excretion rate (for Ricinoleic Acid) ranging from 1% as 5% of the dose of Ricinoleic Acid was detected in the dosed to 4%. Following oral administration for up to 30 days, the ac- skin, and as much as 16% of the dose was detected in the stratum

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48 COSMETIC INGREDIENT REVIEW corneum with mineral oil formulations. With polyethylene gly- approximately 450 µg/cm2/h at 1 h. Mean values for the TEWL col (PEG) formulations, the amount of Ricinoleic Acid remain- rate were approximately 400 µg/cm2/h at 1 h and 250 µg/cm2/h ing in the skin tissues was considerably less. The cutting fluid at 24 h. Additionally, castor oil was the only substance that ex- additives significantly decreased Ricinoleic Acid partitioning hibited a burst effect, i.e., initially, the membrane became fully from the formulation into the stratum corneum in PEG-based hydrated and swollen and was more permeable, providing an mixtures. Additionally, the individual additives or combinations “increased push” effect to pass water through the membrane of these additives significantly reduced Ricinoleic Acid per- toward the anhydrous desiccant. meability in silastic membranes and porcine skin (Baynes and Riviere 2004). Effect on Smooth Muscle Castor Oil and Ricinoleic Acid Skin Penetration Enhancement Ricinoleic Acid Mathias et al. (1978) evaluated the effect of Castor Oil and Ricinoleic Acid on smooth muscle of the rabbit small intes- In a study by Song et al. (2001), the effects of cis-9- tine. Anesthetized male New Zealand white rabbits (weights octadecenoic acid (oleic acid) and a group of chemically re- between 1.5 and 3.0 kg) were used in ileal loop studies. Cas- lated cis- (Ricinoleic Acid) and trans- (ricinelaidic acid) 12- tor oil (dose = 0.85 ml/kg) was administered into the orad end monohydroxylated derivatives and their ethyl and methyl esters of the ileal loop, and myoelectric recording techniques were on the skin penetration of model hydrophobic (hydrocortisone) used. Ricinoleic Acid was administered by intraluminal infusion and hydrophilic (5-fluorouracil) drugs were evaluated in vitro us- (2 µg/kg/min [6 mM] into the orad end). The term migrating ing hairless mouse skin. Test solutions were prepared by placing action potential complex (MAPC) was used to describe the my- either test compound (5-fluorouracil, 5.4 mg; hydrocortisone, oelectric activity that has been observed in certain abnormal 15 mg) with or without an unsaturated fatty acid (50 mg each) states. MAPC has been defined as action potential discharge ac- or fatty acid ester (50 mg each) in a screw-capped glass vial, tivity occurring for longer than 2.5 s, on at least two consecutive and adding propylene glycol (final volume = 1 ml). The mix- electrode sites at 2.5-cm intervals, and having a propagation ve- ture was sonicated to effect complete dissolution. Permeation locity of =∼1.0 cm/s. The myoelectric activity of castor oil and studies were conducted using Vlia-Chien diffusion cells and Ricinoleic Acid was similar to the MAPC that was observed full thickness abdominal skin from female hairless HRS/J mice in the cholera enterotoxin-infected loops. Castor oil induced an (6 weeks old). Ricinoleic Acid enhanced the in vitro transdermal alteration in myoelectric activity. permeation of 5-fluorouracil. In experiments that were designed to study the motor re- The transdermal penetration rate of hydroquinone from sponse in segments of the duodenum, mid-jejunum, terminal propylene glycol was very slow. However, in the presence of ileum, and sigmoid colon (four experiments, respectively), Ri- oleic acid, the penetration rate accelerated and was enhanced cinoleic Acid was diluted with 0.9% sodium chloride solution to approximately 1800-fold. In the presence of Ricinoleic Acid, avolume that would deliver 1.14 ml/h. No MAPC activity was the enhancement of hydroquinone skin penetration was less than observed in control experiments in which 0.9% sodium chloride 1.5-fold. Regarding the penetration of 5-flurouracil, fluxes were solution was perfused into the duodenum at a rate of 1.14 ml/h. much higher in the presence of oleic acid (333-fold increase; When compared to control values, there was no statistically sig- p <.001) than in the presence of Ricinoleic Acid (<5-fold nificant difference in the slow wave frequency or slow wave increase) (Song et al. 2001). propagation velocity in loops exposed to castor oil or Ricinoleic Acid (Mathias et al. 1978). Transepidermal Water Loss Lodge (1994) evaluated the potential for Ricinoleic Acid to Castor Oil induce vasoconstriction using tissues from male New Zealand Lieb et al. (1988) evaluated transepidermal water loss white rabbits. After removal of the thoracic aorta and left and (TEWL) using a modification of a flow-through diffusion right external jugular veins, the vessels were cleaned and cut cell that was originally designed by Bronaugh and Stew- into rings. Following removal of the endothelium, the rings art (1985). The membrane was described as full-thickness, were suspended between two wire holders in tissue baths. Ri- cartilage-stripped skin obtained from the ventral ear of the male cinoleic Acid was diluted with ethanol to produce final bath Syrian golden hamster. Castor oil (10 µl) was placed on the skin concentrations ranging from 0.1 to 30 µg/ml (molar concen- (0.32-cm2 area), and a TEWL rate–versus-time study was con- tration range of 0.335 to 100.50 µM). Ricinoleic Acid in- ducted. Tritiated water (HTO) permeated through the membrane duced vasoconstriction (EC50 = 0.24 ± 0.04 µg/ml) that was into the diffusion cell’s receptor compartment. Flux (TEWL, concentration-dependent and sensitive to the inhibitory effect µg/cm2/h) was determined (using a scintillation counter) by of ifetroban, but not indomethacin. Ricinoleic acid also evoked measuring the HTO that adsorbed to anhydrous calcium chlo- concentration-dependent force development (constriction) in the ride in the receptor cell. Castor oil caused a marked decrease in aorta (EC50 = 4.7 ± 0.7 µg/ml), and this response was antago- the TEWL rate. For untreated skin, the mean TEWL rate was nized by ifetroban.

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CASTOR OIL 49

Sodium Ricinoleate, Castor Oil, and Ricinoleic Acid intraplantar injection with carageenan was preceded by repeated Stewart and Bass (1976a) evaluated the effects Sodium Rici- (8 days) topical applications of Ricinoleic Acid (900 µg/mouse), noleate, castor oil, and Ricinoleic Acid on the digestive contrac- again, a marked prolongation of paw withdrawal latency to heat, tility of the canine small bowel using four male, mixed breed compared to the vehicle control group, was noted. dogs (weights between 10 and 15 g). After feeding, each animal Another experiment involved the repeated local application (8 received a single bolus infusion of Sodium Ricinoleate (500 mg days) of Ricinoleic Acid (900 µg/mouse) or the repeated (4 days) in 30 ml of saline) into the duodenum. Saline (30-ml volume) intradermal injection of Potassium Ricinoleate (30 µg/mouse) served as the control. A duodenal cannula was implanted in each on the ventral surface of the right paw. Freund’s adjuvant had animal, and circular muscle activity was recorded for 1 h. Each been injected into the hindpaw on the first day. The paw with- animal was dosed with Sodium Ricinoleate in two experiments, drawal latency to a painful stimulus (heat) after topical applica- and with isotonic saline (30 ml) in two control experiments. In tion was increased over at least 2 weeks. The intradermal injec- additional experiments, alterations in digestive contractile pat- tion of Potassium Ricinoleate induced a significant antinocicep- terns were monitored after administration (gastric tube) of castor tive effect that lasted for at least 3 weeks (Vieira et al. 2000). oil or Ricinoleic Acid (volume = 10 ml) one h after feeding. Each treatment was repeated twice in each of two animals. Intraduodenal administration of Sodium Ricinoleate pro- Effect on Enzyme Activity duced a biphasic response in the proximal and mid-jejunum con- Ricinoleic Acid sisting of brief initial stimulation, followed by inhibition of di- In a study by Gaginella et al. (1978), isolated villus cells gestive contractility. Compared to the control experiments, these from the small intestine of the hamster were used to deter- stimulatory and inhibitory effects on intestinal contractility were mine whether Ricinoleic Acid stimulates intestinal adenylate described as significant (p <.05) changes in the average force cyclase. In the adenylate cyclase assay, activity of the enzyme per contraction. The changes in mid-jejunal digestive contrac- was determined using a method that is based upon the conver- tility were similar to those observed in the proximal jejunum. sion of [α−32P]ATP to [32P]cyclic AMP. The cell homogenates The oral administration of Ricinoleic Acid (10 ml) induced (30 to 50 µgofprotein per tube) were incubated for 20 min. − − alterations in digestive patterns that were qualitatively similar Ricinoleic Acid concentrations of 10 9 to 5 × 10 3 M did not to those induced by the intraduodenal administration of Sodium cause dose-dependent stimulation of adenylate cyclase. Signifi- Ricinoleate. Brief initial stimulation, followed by an apparent cant (p <.05), but slight, stimulation of adenylate cyclase was − depression of intestinal contractility, was observed after oral noted at a concentration of 10 5 M, and higher concentrations dosing with Castor Oil (10 ml) (Stewart and Bass 1976a). inhibited basal activity. Prostaglandin E1, which is chemically similar to Ricinoleic Acid, was a potent stimulant of adenylate <. Sodium Ricinoleate and Methyl Ricinoleate cyclase activity, causing dose-related stimulation (p 001) of −9 −4 In a study by Stewart et al. (1975), Sodium Ricinoleate de- adenylate cyclase at concentrations of 10 to 10 M. pressed the smooth muscle twitch response of the electrically Simon and Kather (1980) studied the modulation of adenylate driven guinea-pig ileum preparation at concentrations ranging cyclase and cAMP-phosphodiesterase, key enzymes of cAMP from 1.25 × 10−5 to 4 × 10−4 M. Methyl Ricinoleate failed to metabolism, by Ricinoleic Acid in human colonic mucosa. depress the smooth muscle twitch response over this range of test The experiments were performed using histologically normal concentrations. Similar results were reported for spontaneously colonic mucosa obtained from eight patients (five males, three contracting rabbit jejunum. females) who were undergoing hemicolectomy for carcinoma or diverticulosis. Adenylate cyclase activity was determined according to the method of Salomon et al. (1974), and cyclic Antinociceptive Activity AMP phosphodiesterase activity was determined according to Ricinoleic Acid and Potassium Ricinoleate the method of Pöch(1971). Ricinoleic Acid was tested at con- Vieira et al. (2000) evaluated the antinociceptive activity of centrations ranging from 1 × 10−8 to 5 × 10−4 mol/l. Basal Ricinoleic Acid (in peanut oil), dissolved in a vehicle contain- adenylate cyclase activity in homogenates of human colonic mu- ing 10% ethanol, 10% Tween 80, and 80% saline, using groups cosa averaged 250 pmol cAMP/mg protein/15 min. Mean basal of 5 male albino Dunkin-Hartley guinea pigs (weights = 250 cAMP phosphodiesterase activity was 150 ± 15 pmoles per mg to 350 g) and groups of 8 to 10 male Swiss mice (weights = protein per minute. 20 to 25 g). A series of experiments was performed. In the first Over the range of concentrations tested, dose-related stimu- experiment, a single topical application of Ricinoleic Acid (900 lation of the large bowel adenylate cyclase was not observed. µg/mouse) was made to the ventral surface of the right paw of Basal activity of adenylate cyclase was inhibited at concentra- each mouse. Test substance application was followed by intra- tions above 1 × 10−5 mol/l. Ricinoleic Acid did not influence plantar injection with carrageenan (to induce edema) 30 min soluble cAMP phosphodiesterase activity over the range of con- later. Ricinoleic acid significantly reduced the reaction time to centrations tested. Ricinoleic Acid was an ineffective stimulus of a heat stimulus, compared to treatment with the vehicle. When human colonic adenylate cyclase, and also was not a competitive

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50 COSMETIC INGREDIENT REVIEW inhibitor of soluble cAMP phosphodiesterase activity (Simon occurred between either of the dietary groups (with or without and Kather 1980). cholesterol in the diet). In a study by Vanderhoek et al. (1980), the effect of Rici- Compared to the high-oleic safflower oil dietary group (with noleic Acid on the oxygenation of [1-14C]arachidonic acid by or without cholesterol), significantly decreased (p <.05) hep- platelet cyclooxygenase and lipoxygenase enzymes was moni- atic triacylglycerol concentrations were reported for the cas- tored using an oxygen electrode and by analysis of the radioac- tor oil dietary group. For cholesterol-enriched diets, the castor tive products formed. The enzymes were obtained from human oil and coconut oil dietary groups had significantly increased platelet concentrates (3 to 4 days old). Ricinoleic Acid did not (p <.05) hepatic cholesterol when compared to the high-oleic appear to have an effect on either enzyme. The concentration of safflower oil dietary group. It was concluded that castor oil had a Ricinoleic Acid that was required for half-maximal inhibition hypocholesterolemic effect in rats in this study (Ihara-Watanabe of lipoxygenase or cyclooxygenase activity was >300 µM. et al. 1999). Beubler and Schrirgi-Degen (1992) studied the stimulation of enterocyte protein kinase C by laxatives in vitro us- Effect on Leukocyte Infiltration and ing intestinal epithelial cell (from female Sprague-Dawley rats) Prostaglandin Synthesis preparations. A protein kinase C assay system was used to Castor Oil determine the activity of protein kinase C. This assay sys- Ohia et al. (1992) used castor oil as a vehicle control in a tem is based on protein kinase C–catalyzed transfer of the γ - study evaluating the effects of steroids and immunosuppres- phosphate group of adenosine-5-triphosphate to a peptide that sive drugs on endotoxin-induced uveitis in rabbits. The vehi- is specific for protein kinase C. Ricinoleic Acid (dissolved in cle control group consisted of four male New Zealand rabbits DMSO) stimulated protein kinase C activity in a concentration- (weights = 1.5 to 2.0 kg). The animals were injected intramus- dependent manner within the concentration range of 2 to cularly with castor oil (dose = 2 mg/kg). At 1 h post injec- 200 µg/ml. tion, both eyes of each rabbit were cleansed and anesthetized. This procedure was followed by the intravitreal injection of Es- Effect on Uridine Uptake cherichia coli endotoxin (100 ng). The animals were killed 24 h At a concentration of 50 µM, Ricinoleic Acid suppressed after injection of the endotoxin. Aqueous humor was obtained the uptake of uridine into human diploid fibroblasts in culture and leukocytes were counted. The eyes were then enucleated (Polgar and Taylor 1977). for microscopic examination. Compared to saline-treated controls, castor oil significantly reduced the infiltration of leuko- Effect on Lipid Metabolism cytes and the prostaglandin E2 (inflammatory mediator) content Castor Oil of the aqueous humor and iris-ciliary body. The authors concluded that Castor Oil had an anti-inflammatory effect in this Ihara-Watanabe et al. (1999) evaluated the effect of the fol- study. lowing diets on lipid metabolism using groups of five male Wis- tar rats (3 weeks old): 10% castor oil, 10% high-oleic safflower oil, and 10% coconut oil. The diets were prepared in the absence Effect on Cytokine-Induced Endothelial Cell Activation or presence of dietary cholesterol, and were fed ad libitum for Sodium Ricinoleate 20 days. On day 20, blood samples were obtained and liver and De Caterina and Bernini (1998) evaluated the relationship perirenal adipose tissues were excised. Serum and hepatic lipids between structural differences in fatty acids and the inhibition were analyzed for total cholesterol, high-density lipoprotein, tri- of endothelial activation, taking into consideration that dietary acylglycerols, and phospholipids. long-chain fatty acids may influence processes that involve en- Body weight gain and food intake were lowest for rats fed dothelial activation (e.g., inflammation and atherosclerosis). To the castor oil diet enriched with cholesterol, but not for rats fed test for a potential stimulatory effect of fatty acids on adhesion cholesterol-free diets. Body weight gain to consumption ratios molecule expression, sodium salts of saturated, monounsatu- were similar among all of the groups. Compared to rats fed the rated (Sodium Ricinoleate included), and n-6 and n-3 polyun- high-oleic safflower oil diet (enriched with cholesterol), rats fed saturated fatty acids were each incubated alone (test concentra- the castor oil diet enriched with cholesterol had decreased liver tion = 25 µM) with human saphenous vein endothelial cells weights (g/100 g of body weight). Data on liver weights were (HSVECs) for 72 h. The 72-h incubation period was followed not presented in the study. by 16 h of stimulation with the cytokine interleukin (IL)-1α in Values for total cholesterol and phospholipids in the serum the continuous presence of fatty acid salt. At the end of the in- were significantly decreased in rats fed castor oil, compared to cubation period, vascular cell adhesion molecule-1 (VCAM-1) rats fed high-oleic safflower oil in the absence or presence of expression was assessed using a cell surface enzyme immunoas- cholesterol. For cholesterol-enriched diets, the lowest value for say (EIA). serum high-density lipoproteins was reported for the castor oil Sodium Ricinoleate significantly inhibited (considered mod- dietary group. No significant differences in serum triacylglycerol est; p <.05) cytokine-induced endothelial adhesion molecule

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CASTOR OIL 51 expression. DHA inhibited VCAM-1 expression (p <.01) most For the trypan blue exclusion method, Sodium Ricinoleate potently (De Caterina and Bernini 1998). caused dose-related increases in cytotoxicity at concentrations De Caterina and Bernini (1998) also conducted an experiment of 0.1 to 5.0 mM. Death of all cells was noted at a concentration to determine whether the fatty acid sodium salts may induce en- of 2.0 mM. Results for control incubation in the 51Cr release as- dothelial activation in the absence of cytokines. The salts were say indicated that 26.8 ± 1.7% of 51Cr, initially within or bound incubated with endothelial cell monolayers for 24 h. Incubation to the cells, was released into the medium. Sodium Ricinoleate periods prior to detection of adhesion molecule surface expres- induced concentration-dependent (0.1 to 5 mM Sodium Rici- sion varied from 0 to 72 h. Neither Sodium Ricinoleate (25 µM) noleate) increases in 51Cr release, from 4.2% to 54.8% above the nor any other fatty acid sodium salt induced adhesion molecule control value. In the 3-O-methylglucose uptake assay, Sodium expression, as assessed by EIA. Ricinoleate inhibited the uptake of 3-O-methylglucose over a concentration range of 0.1 to 2.0 mM (Gaginella et al. 1977b). Ricinoleic Acid In a study by De Caterina and Bernini (1998), the cytotoxi- In a study by De Caterina et al. (1995), the effect of Rici- city of Sodium Ricinoleate and other fatty acid sodium salts in noleic Acid on VCAM-1 expression was evaluated using rabbit human saphenous vein endothelial cell cultures was evaluated. 3 coronary artery endothelium in vitro. VCAM-1 (marker for en- Cell count, trypan blue exclusion, and [ H]leucine incorporation dothelial cell activation) is a surface protein that is not expressed into total cell-associated and released proteins were monitored. by endothelial cells in vitro, but is inducible by exposure to in- For the latter of the three, trichloroacetic acid precipitation of flammatory stimuli such as bacterial endotoxin, IL-1, or tumor total cell extracts and cell supernates was performed. At con- necrosis factor α (TNFα). Ricinoleic Acid significantly reduced centrations ≤25 µM, Sodium Ricinoleate and other fatty acid IL-4–induced VACM-1 expression (50% inhibitory concentra- sodium salts did not induce significant toxicity (based on cell tion between 10 and 100 µM). Neither cell number nor viability count and trypan blue exclusion) for periods up to 72 h prior to was increased in this concentration range. Ricinoleic Acid did cytokine stimulation. not inhibit TNFα-induced VACM-1 expression to any significant extent. Antimicrobial Activity Castor Oil Effect on Prostaglandin Synthesis Morris et al. (1979) evaluated the antimicrobial activity of Castor Oil castor oil on Staphylococcus aureus, Escherichia coli, and Can- Gao et al. (1999) evaluated the effect of castor oil in the diet dida albicans using the Petri plate–paper disc procedure. Paper µ on the synthesis of prostaglandin E2 (PGE2) and the induction of discs were soaked with 20 lofa10% Castor Oil solution, and labor using two groups of eight pregnant Wistar rats (at gestation the discs were immediately applied to solidified, seeded agar day 18; test and control groups, respectively). Two milliliters of (1 disc per plate). The plates were incubated for 10 to 24 h. castor oil–containing diet were administered by gavage daily for Discs containing 95% ethanol (20 µl) served as negative con- a total of four feedings. The diet (labor-inducing diet) consisted trols. Castor Oil did not have antimicrobial activity on any of of castor oil (30 ml) + one chicken egg, blended and heated to the strains tested. a thick consistency. At 4 h after the fourth feeding, the pregnant females were killed. Compared to the control group, a significant Sodium Ricinoleate increase in concentrations of PGE2 in tissues of the intestinal mucosa, placenta, amnion, and amniotic cells was noted in test Whiteside-Carlson et al. (1955) studied the effect of Sodium Ricinoleate on cell division using a laboratory strain of E. coli. animals. The authors stated that the increased synthesis of PGE2 “is a key of” the initiation of labor that is induced by a castor oil Sodium Ricinoleate was tested over a concentration range of diet. 1to5mg/ml of medium. The cultivation of E. coli on nutri- ent agar that was saturated with Sodium Ricinoleate caused the development of filamentous forms during the first h of loga- Cytotoxicity rithmic growth. By the end of the log phase, organisms of nor- Sodium Ricinoleate mal morphology were observed. When agar containing Sodium Gaginella et al. (1977b) evaluated the cytotoxicity of Rici- Ricinoleate was adjusted to pH values of 5 to 8, filament for- noleic Acid using epithelial cells that were isolated from the mation was more marked and persisted for longer periods at small intestines of male Syrian hamsters. Cytotoxicity was as- the more alkaline pH value. The utilization of liquid proteose- sessed by exclusion of trypan blue, the release of intracellular peptone medium (with periodic readjustment of the pH to a (prelabeled) 51Cr, and inhibition of the cellular uptake of 3- range of 7.6 to 7.8) resulted in maintenance of the cells in fila- O-methylglucose. The results produced by all three methods mentous form throughout the entire growth cycle. The tendency indicated that Sodium Ricinoleate produced a dose-dependent toward filament formation was reduced by lowering the incuba- cytotoxicity. tion temperature to below 37◦C. The results of this study indicate

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52 COSMETIC INGREDIENT REVIEW that Sodium Ricinoleate interfered with cell division in E. coli, increased the intraluminal release of acid phosphatase (AP) in causing the organisms to develop as filaments. the duodenum and jejunum (p <.01), and a similar trend was Mordenti et al. (1982) studied the activity of Sodium Rici- reported for the ileum and colon (p < 0.2to.1). A correlation noleate against Streptococcus mutans strain 6715-13 (plaque) in between increased release of AP and intestinal hyperemia was vitro. The minimum inhibitory concentration of Sodium Rici- established. The authors noted that the results of this study sug- noleate against S. mutans was1%(or 3.12 × 10−2 M). During gest that PAF is involved in the mechanism underlying castor testing for the minimum inhibitory concentration, the following oil–induced intestinal damage (Pinto et al. 1989). four distinct concentration-dependent cell growth and acid pro- In a study by Blair et al. (2000), the affinity of castor oil and duction phenomena were identified: (1) no cell growth, pH > numerous other chemicals for the estrogen receptor (from uteri 6.8; (2) cell growth, medium pH > 6.8; (3) cell growth, medium of ovariectomized Sprague-Dawley rats) was evaluated using a pH of 5.2 to 6.8; and (4) cell growth, medium pH < 5.2. It validated estrogen receptor competitive binding assay. In this was concluded that Sodium Ricinoleate had bactericidal activity assay, the ability for a chemical to compete with [3H]estradiol 3 in this assay. In subsequent experiments, wire-adherent plaque ([ H]E2) for the estrogen receptor was determined, based on IC50 specimens were treated with various concentrations of Sodium (concentration resulting in 50% inhibition of [3H]estradiol bind- Ricinoleate. The intact plaque samples survived in each of these ing) values. All assays were replicated a minimum of two times. −4 tests. The authors stated that the ability for Sodium Ricinoleate The mean IC50 for Castor oil was >1.0 × 10 M. Mean IC50 val- to kill bacteria in suspension, but not in intact plaque, clearly ues for 4-heptyloxyphenol and phenophthalein (both classified demonstrates the absence of a correlation between antibacterial as moderate estrogen receptor binders) were 6.75 × 10−5 ± activity against cells in suspension and antiplaque activity. 0.75 × 10−5 and 6.73 × 10−6 ± 1.79 × 10−6, respectively. The binding affinity of castor oil for the estrogen receptor was not classified. Other Biological Effects Castor Oil Ricinoleic Acid Capasso et al. (1986) evaluated the effect of castor oil on In a study by Grainger et al. (1982), the effect of Ricinoleic the formation of histamine, 5-hydroxytryptamine (5-HT), and Acid on lymph flow in outerperfused segments of cat ileum was prostaglandin-like material (PG-LM) in the rat intestine using evaluated. The cats were anesthetized and a segment of ileum male, fasted Wistar-Nossan rats (number and weights not stated). with intact innervation and lymphatic drainage was isolated and Castor oil (dose = 2 ml/rat) was administered by gavage. Follow- perfused by the intact mesenteric artery. A cannula was inserted ing the onset of diarrhea, the animals were killed, and sections of into a large lymphatic vessel emerging from the mesenteric pedi- colon removed. Untreated rats served as controls. Castor oil in- cle, and lymph flow was determined by observing lymph move- duced an approximately fourfold increase in PG-LM (p <.01) ment in a calibrated pipette that was connected to the lymphatic and a threefold increase in histamine and 5-HT (p <.01). A cannula. The mean control (± SE)value for intestinal lymph decrease in the Castor oil–induced colonic formation of PG- flow in the experiments was 0.048 ± 0.005 ml/min × 100 g. LM was noted following pretreatment of the animals with in- Net fluid secretion was stimulated by intraluminal instillation of domethacin (inhibitor of prostaglandin biosynthesis) or hydro- 5mMRicinoleic Acid. The peak increase in intestinal lymph cortisone. It was concluded that these data support the idea that flow produced by Ricinoleic Acid in all experiments was 6.3 the laxative effect of castor oil is the result of increased intestinal ± 1.2 (n = 5) times the control. The maximal fluid excretion production of PG-LM, histamine, and 5-HT. rate observed with Ricinoleic Acid was 0.43 ± 0.23 ml/min × A study was conducted by Pinto et al. (1989) to determine 100 g. whether oral administration of castor oil leads to changes in Yagaloff et al. (1995) evaluated the inhibitory activity of a se- the levels of platelet-activating factor (PAF) formed by intesti- ries of fatty acids and fatty acid derivatives on [3H]leukotriene nal tissue, and whether these changes are related to intestinal B4 binding (LTB4)topig neutrophil membranes using the LTB4 damage. Fasted, male Wistar-Nossan rats (number not stated; receptor binding assay. The most potent chemicals were 15- = = weights 130 to 140 g) were used. Castor oil (dose 2 ml/rat) hydroxy-DGLA (20:3; Ki = 1 µM), eicosadienoic acid (20:2; was administered orally, and the animals were killed. Ki = 3 µM), and Ricinelaidic Acid (18:1; Ki = 2 µM). Rici- At 3 h post dosing, the formation of PAF by segments of noleic Acid (18:1) had a Ki of 9 µM. rat intestine was significantly increased (compared to control rats treated with olive oil) in the following intestinal segments: <. <. <. duodenum (p 001), jejunum (p 01), ileum (p 2), and ANIMAL TOXICOLOGY colon (p <.1). At macroscopic examination, extensive regions of hyperemia were noted in the entire intestinal mucosa. The Acute Oral Toxicity duodenum and jejunum regions were the most severely affected. Castor Oil No increase in PAF was noted in control rats dosed with olive oil. In a study by Capasso et al. (1994), castor oil (2 ml) was Compared to control rats dosed with olive oil, castor oil also administered orally to ten male Wistar rats (weights = 160 to

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CASTOR OIL 53

180 g). The animals were killed and two segments from stan- Hydrogenated Castor Oil dardized regions of the duodenum and jejunum were visibly The acute oral toxicity of a 25% solution of Hydrogenated evaluated for macroscopic damage. Mucosal injury was graded castor oil in olive oil was evaluated in a study involving ten male according to the following scale: 0 (normal), 1 (hyperemia), 2 Wistar rats (average body weight = 155 g). The test solution (hyperemia with evidence of hemorrhage into the lumen), and 3 (dose = 10 g/kg) was administered via a gastric probe. Toxic (severe hemorrhage into the lumen). signs were not observed in any of the animals, indicating that Copious diarrhea was reported for all animals on days 3, 5, the test substance was well tolerated. The LD50 was >10 g/kg and 7 post dosing. Macroscopic damage, characterized mainly (Allegri et al. 1981). by vasocongestion, was observed throughout the duodenum and jejunum. The injury observed ranged from mild (at 1 h) to severe Ricinoleic Acid (at 5 h), and was less severe at 7 h. Injury was not observed at In a study by Okui et al. (1964), male albino rats (number 0.5 or 9 h after dosing. Castor oil–induced mucosal damage was not stated; weight = 120 ± 10 g) were injected intraperitoneally associated with statistically significant intraluminal release of (i.p.) with 100 mg Ricinoleic Acid. All of the animals were acid phosphatase. dead within 24 h. The i.p. injection of a 5% Ricinoleic Acid In additional experiments (groups of 10 rats), mucosal dam- emulsion (1 ml) was tolerable, but caused an increase in the fol- age induced by castor oil was exacerbated in the presence of a ni- lowing: amount of ascites, conglutination of peritoneal organs, ω tric oxide synthase inhibitor (N -nitro-l-arginine methyl ester), and swelling of the liver. However, histological changes were suggesting that nitric oxide provides protection against castor not observed in liver specimens. oil-induced mucosal damage (Capasso et al. 1994). In a study involving male Crl:CD BR rats, the findings sug- Cetyl Ricinoleate gested that castor oil–induced diarrhea is the result of activation In an acute oral toxicity test performed by Laboratoire de of NK1 and NK2 receptors by endogenous tachykinins (Croci Recherche et d’Experimentation (1994), Cetyl Ricinoleate was et al. 1997). evaluated using five female NMRI EOPS mice (weights = 19 to In a study by Johnson et al. (1993), castor oil was ad- 22 g). Oral dosing (single dose 2000 mg/kg) was followed by a ministered orally to 12 ponies (17 months to 20 years old; 6-day observation period. None of the animals died, and clinical = weights 160 to 250 kg). The ponies were randomly as- and behavioral evaluations were normal. Weight gain was con- signed to two treatment groups and one control group (four sidered satisfactory. The authors stated that Cetyl Ricinoleate ponies per group). Each animal received a single 2.5 ml/kg was non-toxic in this study. body weight dose of castor oil via nasogastric tube. Diarrhea occurred within 24 h of dosing. The animals were killed and Ethyl Ricinoleate necropsied at 24, 48, and 72 h post dosing. Intestinal tis- The acute oral toxicity of Ethyl Ricinoleate was evaluated us- sues were removed and subjected to gross and microscopic ing 10 rats (weights and strain not stated). The acute oral LD50 examination. exceeded 5.0 g/kg; one animal receiving this dose died. The In ponies killed at 24 or 48 h, gross lesions were most severe following clinical signs were observed during the study: iso- in the cecum and ventral colon. At 24 h, the mucosa of the dis- lated instances of diarrhea, chromorhinorrhea, ptosis, and chro- tal jejunum, ileum, cecum, and ventral colon was diffusely red. modacryorrhea. At necropsy, gross observations were normal for Most of the ponies dosed with castor oil had generalized hyper- eight animals. Necropsy of the remaining two animals revealed emia of the intestinal mucosa at 24, 48, and 72 h post dosing, the following changes in either one or both animals: red and/or and mesenteric, cecal, and colonic lymph nodes were edema- yellow areas in the intestines, red areas in the stomach, mottled tous. Mucosal and submucosal edema of the ileum, cecum, and liver, mottled spleen, mottled kidneys, dark lungs, red exudate ventral colon was noted in all ponies dosed with castor oil. At in the anogenital area, and a large amount of blood in the bladder 72 h post dosing, focal acute ulceration and intense mucosal hy- (Research Institute for Fragrance Materials [RIFM] 2000). peremia of the glandular stomach was observed in three of four ponies dosed with castor oil. Microscopic changes in the superficial enterocytes of the ce- Acute Dermal Toxicity cum and ventral colon that were characterized as follows: loss Ethyl Ricinoleate of microvilli; distortion of the cytoplasmic terminal web; ex- The acute dermal toxicity of Ethyl Ricinoleate was evaluated pansion of the cytoplasmic matrix, with formation of precipi- using six rabbits (weights and strain not stated). The acute dermal tates; and widening of intracellular spaces between junctional LD50 exceeded 5.0 g/kg; one animal receiving this dose died. complexes. Castor oil induced acute superficial enterocolitis The following clinical signs were observed during the study: in ponies. Dosing was associated with transient diarrhea, neu- isolated instances of lethargy, diarrhea, ptosis, and yellow nasal trophilic inflammation, and mucosal erosion in the cecum and discharge, and moderate erythema (six rabbits), slight edema ventral colon (Johnson et al. 1993). (one rabbit), and moderate edema (five rabbits). At necropsy,

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54 COSMETIC INGREDIENT REVIEW gross observations were normal for three animals. Necropsy of Using electron microscopy, parenchymal cells containing the remaining animals revealed the following changes: red areas electron-dense lipid inclusions were observed in the zona fas- in the intestines, mottled liver, white nodules in the liver, dark ciculata of adrenal glands from mice injected with castor oil. areas in the lungs, mottled kidneys, pale kidneys, dark spleen, By light microscopy, there was no difference between adrenal and dark areas in the stomach (RIFM 2000). glands from control mice and mice injected with the vehicle consisting of castor oil and benzyl benzoate; however, by electron Acute Intravenous Toxicity microscopy, the following variations were observed: profound Castor Oil disruption of parenchymal mitochondria in the zona fasciculata In a study by Lorenz et al. (1982), castor oil was administered and a noticeable increase in size for some of the mitochondria. intravenously (single bolus dose = 0.1 ml/kg) to each of eight The vesicular orientation of the inner mitochondrial membrane anesthetized adult mongrel dogs (four males, four females; mean was lost (i.e., random orientation), and diminution of the mito- weight = 20.5 kg). Blood for histamine assays was obtained chondrial matrix was also observed. at 2, 5, 10, 15, and 20 min post dosing. The incidence of the Unlike the controls, the macrophages of exposed animals following signs was determined: erythema, hives (papules and had large vacuoles filled with globular material. In the zona wheals), edema, defecation, tachypnea, hypotension, and death reticularis and inner zona fasciculata, parenchymal mitochon- (circulatory arrest). None of the animals died, and erythema dria with circular orientation of the inner membrane (normally (one animal; score not stated) was the only clinical sign that was few) were noticeably increased in number. The authors noted reported. An increase in blood histamine occurred in one of the the appearance of similar mitochondria following adrenocorti- eight animals. At 2 min post dosing, the increase was 0.6 ng/ml cotrophic hormone (ACTH) stimulation, and that the alteration of whole blood (maximum response) in this animal. The authors of ACTH release has been shown to result from stress. The concluded that castor oil was ineffective in terms of its ability author concluded that subcutaneous injection of the castor oil– to cause histamine release. benzyl benzoate vehicle caused morphological alterations that are suggestive of stress-induced changes (Migally 1979). Short-Term Oral Toxicity Castor Oil Subchronic Oral Toxicity Masri et al. (1962) conducted an experiment in which ten Castor Oil = ± male weanling rats (4 weeks old; mean weight 42.2 0.6 g) In a study by the National Toxicology Program (NTP) (1992), were fed (ad libitum) 10% castor oil for approximately 5 weeks. the subchronic oral toxicity of castor oil was evaluated using = ± A similar control group (mean weight 42.2 0.5 g) re- groups of 10 male and 10 female F344/N rats (mean body ceived corn oil in the diet. The animals were observed for weights = 126 to 132 g [males]; 107 to 110 g [females]) and growth and catharsis throughout the duration of the study. Cas- groups of 10 male and 10 female B6C3F1 mice (mean body tor oil in the diet did not induce catharsis in any of the 10 weights = 22.6 to 23.0 [males]; 17.2 to 17.7 [females]). In the rats, and growth of these animals was comparable to that ob- core study, five groups of test animals (mice or rats) received served in the control group. At the end of the experiment, diets containing 0.62%, 1.25%, 2.5%, 5.0%, or 10% castor oil ± mean weights for test and control rats were 155.6 4.6 and continuously for 13 weeks. Two control groups of 20 rats and ± 160.5 4.1 g, respectively. Grossly, there were no abnormal- 20 mice received diets that did not contain castor oil. ities. Additionally, no abnormalities of the perirenal fat were Ten additional rats/sex were included at each dose level for noted. evaluation of hematological and clinical chemistry parameters. On days 5 and 21, these animals were anesthetized with CO2 and Short-Term Subcutaneous Toxicity blood samples were obtained. These animals were killed follow- Castor Oil ing blood collection on day 21. Blood samples for hematology In a study by Migally (1979), the morphology of the adrenal and clinical chemistry were also collected from core-study rats cortex was examined after injection of a commonly used vehicle at the end of the study. Also, at the end of the 13-week study, all consisting of castor oil and benzyl benzoate. Twenty-five adult core-study animals were killed and complete necropsies were C57 BL/6J mice (weights not stated) were divided into the fol- performed, including complete microscopic examination on tis- lowing groups: group I (five intact control mice); group II (five sues, on all test and control animals. sham-injected mice); group III (five mice each injected with 0.1 No effect on survival or significant differences in average ml castor oil); group IV (five mice each injected with 0.1 ml food consumption were noted in any of the five groups of benzyl benzoate; and group V (five mice each injected with 0.1 male and female rats fed castor oil. There were no signifi- ml castor oil and benzyl benzoate [4:1]). The mice were injected cant differences in mean body weights between test and control subcutaneously daily for four weeks and then killed. Necropsy groups. was then performed and tissues prepared for light and electron The following hematological effects were reported for microscopic examination. male rats: a slight decrease in mean corpuscular hemoglobin

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CASTOR OIL 55 concentration (MCHC) (10% castor oil diet), a statistically To confirm the authors’ explanation of the results, the exper- significant decrease (p <.05) in mean corpuscular volume imental trial was repeated with Hydrogenated Castor Oil at con- (MCV) (10% castor oil diet), a decrease in mean corpuscular centrations of 1%, 5%, and 10% in corn oil (effective concentra- hemoglobin (MCH) (5% and 10% castor oil diets), and an in- tions of 0.865%, 4.33%, and 8.65%, respectively). The amount crease in platelets (1.25%, 5%, and 10% castor oil diets). The of corn oil added to the individual diets was 9%, 15%, and 10%. only hematological abnormality reported for female rats was a Corn oil (20%) was present in the control diet. Necropsy was statistically significant (p <.05) decrease in reticulocyte counts performed at the end of the 90-day feeding period. The growth (0.62% or 10% castor oil diets). The authors did not consider rate for rats fed 8.65% Hydrogenated Castor Oil in the diet was the hematological effects to be biologically significant. A dose- equivalent to that of the other dietary groups. Based on organ related increase (treatment-related) in the activity of serum al- weight determinations and the results of hematological and mi- kaline phosphatase was noted in male and female rats at days 5 croscopic evaluations, no adverse effects were observed (Binder and 21, and at the end of the study. et al. 1970). Increased heart-to-body weight ratios were reported for male In the 16-week feeding study (Binder et al. 1970), groups of rats that received 0.62%, 2.5%, and 10.0% castor oil diets; 15 male albino rats (Slonaker substrain of Wistar strain, weights however, no increase in absolute heart weight was noted. The = 43 to 83 g) received diets containing the following oils: group observed differences in organ-to-body weight ratios were not 1 (fed 0.865% HCO + 19% corn oil for 16 weeks), group 2 considered treatment related. Microscopically, no morphologic (0.865% HCO + 19% corn oil for 8 weeks, then 20% corn changes were associated with the slight differences in organ oil for 8 weeks), group 3 (8.65% HCO + 10% corn oil for 16 weights between the dietary groups. weeks), and group 4 (8.65% HCO + 10% corn oil for 8 weeks, Compared to controls, liver weights were increased in male then 20% corn oil for 8 weeks). and female mice receiving diets containing 5% or 10% castor The effective dietary concentrations of Hydrogenated Castor oil. Increased kidney weights were reported for female mice that Oil in the 0.865% and 8.65% diets were 0.865% and 8.65%, received 5% or 10% castor oil in the diet. Microscopically, there respectively. The control group received 20% corn oil in the diet wasnoevidence of morphologic changes that could be been as- for 16 weeks. sociated with the slight differences in organ weights between the At 4, 8, 12, and 16 weeks, the number of animals on the HCO groups tested. Additionally, there was no evidence of treatment- diets that were alive was 33, 30, 12, and 6 rats, respectively. The related lesions in any of the tissues or organs that were examined corresponding number of rats remaining on the control diet was (all dietary groups). 15, 15, 12, and 6. The authors concluded that diets containing up to 10% castor Weight differences due to the different diets at 4 and 8 weeks oil to B6C3F1 mice and F344/N rats was not associated with were not statistically significant at the 95% level. However, at toxicity to any specific organ, organ system, or tissue in this 12 weeks, rats on the 8.65% HCO diet weighed significantly study (NTP 1992). less than rats on the control diet or 0.865% HCO diet. At 16 weeks, the weight gain of rats on the control diet was signif- Hydrogenated Castor Oil icantly greater than that of rats on the 8.65% HCO diet for 16 In a preliminary feeding trial (preparatory to the 16-week weeks or on the 8.65% HCO diet for 8 weeks + control diet for feeding study reported below) by Binder et al. (1970), the sub- the remaining 8 weeks. The weight gain of rats on the 0.865% chronic oral toxicity of 5%, 10%, and 20% Hydrogenated Cas- HCO diet was not significantly different from that of rats on the tor Oil [HCO] (effective concentrations of 4.33%, 8.65%, and control diet. Except for the reduced growth rate in rats receiving 17.3%, respectively) in the diet was evaluated using groups of the 8.65% HCO diet, no adverse effects were observed (Binder three weanling female rats (Slonaker substrain of Wistar strain). et al. 1970). The sample of HCO that was incorporated into the diet contained 86.5% 12-hydroxystearic acid, 10.3% nonoxygenated acids, and Ocular Irritation/Toxicity 3.2% 12-ketostearic acid. The three groups were fed for 90 days, Castor Oil and the control group received diet only. At necropsy, organ Carpenter and Smyth (1946) evaluated the ocular irritation weights were recorded and numerous tissues were prepared for potential of castor oil using albino rabbits. The eyelids were microscopic examination. Prior to necropsy, blood samples were retracted, and undiluted castor oil (0.5 ml) was applied to the obtained for hematological evaluation. center of the cornea. At approximately 1 min post instillation, the A reduced growth rate in rats fed 8.65% and 17.3% Hydro- eyelids were released. Reactions were scored 18 to 24 h later. To genated Castor Oil, respectively, was the only abnormality that determine the score for ocular injury, the individual numerical was noted. The authors stated that it is likely that Hydrogenated scores for each eye were added together and then divided by Castor Oil was poorly digested because of its high melting point. the number of eyes (usually 5). An ocular injury score of 5.0 Therefore, the poor body weight gains may have been due to the represents severe injury, and corresponds to necrosis (covering lower caloric density of the diets containing Hydrogenated Cas- approximately three-fourths of the corneal surface) that is visible tor Oil at concentrations ≥8.65%. only after staining or a more severe necrosis covering a smaller

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56 COSMETIC INGREDIENT REVIEW area. An injury grade of 1 (maximum score possible = 20) was A micrometer eyepiece was inserted into the microscope, and reported for castor oil. epidermal width measurements (25 per individual field; 400× In a study by Guillot et al. (1979), the effects of various oils magnification) were made. Castor oil induced slight reddening (used in cosmetics) on the rabbit eye were assessed according to at the application site. the official French methods. Following the instillation of undi- Microscopically (pigs and guinea pigs), hyperplasia, with an luted castor oil (no ocular rinsing), no corneal involvement was increase in the cell and nuclear density of the basal cell layers, found. However, slight congestion of the iris and conjunctiva was observed. Additionally, widening of the granular layer and was observed. thick keratohyalin granules were observed. Average epidermal Castor oil served as one of the vehicle controls in a study by widths (25 measurements) were as follows at the end of the Behrens-Baumann et al. (1986) investigating the effect of topical 10-day application period: guinea pigs (control, 27.9 µm; test, cyclosporin A on the abraded and intact rabbit cornea. Two castor 94.3 µm) and pigs (control, 26.5 µm; test, 32.3 m) (Meyer et al. oil vehicle control groups of outbred male rabbits were used. In 1976). both groups (10 eyes/group), 10 drops of castor oil were instilled Rantuccio et al. (1981) evaluated the effect of undiluted cas- daily for 3 weeks, and all eyes were examined using a slit-lamp. tor oil on the skin of five female albino rabbits (6 months old; At the end of the 3-week period, the animals were killed and their mean weight = 2500 g). The test substance (0.5 ml) was gently corneas were excised and prepared for scanning and transmis- massaged into depilated skin of the right flank of each rabbit sion electron microscopy. For eyes (no corneal abrasion) treated daily for 30 days. Following each daily application, the test site with Castor oil and examined by scanning electron microscopy, wascovered with a simple protective dressing. Two small skin there were increased numbers of medium dark and dark epithe- punch biopsies were obtained from the right and left (control) lial cells (compared to the normal epithelium). The cell borders flank on days 10, 20, and 30. Macroscopic skin changes included were intact and the number of holes was reduced. By transmis- slight erythema and slight edema. Microscopic examination of sion electron microscopy, corneal endothelial cells were intact. punch biopsies obtained at day 10 revealed “clear-cut” acan- By scanning electron microscopy, an intact cell pattern, with thosis with vacuolization and disorganization of the basal layer. more cell microstructures than normal, for the corneal endothe- These changes were associated with some infiltration of the der- lium was reported. Ultrathin sections appeared normal. The au- mis by mononuclear cells and fibrocytes. After days 20 and thors concluded that castor oil did not damage the rabbit corneal 30, the epithelial changes became progressively more evident. epithelium or endothelium (Behrens-Baumann et al. 1986). However, the appearance of the infiltrate remained unchanged. In a study by Guillot et al. (1979), the effect of various oils that are used in cosmetics on rabbit skin was assessed according Skin Irritation to the official French methods. An open skin irritation test and Castor Oil repeated application test (8 weeks) were performed. The authors Butcher (1951) evaluated the effect of castor oil and other stated that the two samples of undiluted castor oil, as well as 10% substances on rat epidermis using, primarily, 22-day-old albino aqueous castor oil, were well tolerated. or Long-Evans rats or rats of either strain in which the hair In a study by Motoyoshi et al. (1979), the skin irritation po- follicles were in the resting stage; older animals were also used. tential of castor oil (undiluted) was evaluated using six albino A total of 216 rats were used in the experiments. Using dry angora rabbits (average weight = 2.6 kg). The test substance cotton, castor oil was gently swabbed onto a large area of the (0.1 g of liquid or semisolid) was applied for 24 h to two test dorsum (clipped free of hair) daily for 5 days to 2 weeks. At the areas (clipped free of hair) on the dorsal surface of each an- end of the application period, small areas of skin were obtained imal. A plastic collar was wrapped around the neck of each from the backs of treated and control (one rat/litter) rats and animal and remained throughout the application period, after examined microscopically. Castor oil was described as having a which reactions were scored according to the following scale: mild effect on the epidermis. The alteration was mainly confined − (no reddening) to +++(severe reddening, beet redness). At to the stratum granulosum, where the cells were more numerous 30 min, after reactions were scored, the test areas were clipped and the granules were more evident, compared to the litter-mate free of hair and castor oil was reapplied according to the same controls. procedure. A third application of Castor Oil occurred 48 h later, In a study by Meyer et al. (1976), castor oil (undiluted, 4 and reactions were scored. Reactions were scored again at 72 h. drops) was rubbed onto either side of the shaved left flank of Castor oil was severely irritating to the skin of rabbits (score = 3). each of six albino guinea pigs (weights = 300 to 400 g) and The skin irritation potential of Castor oil was also evaluated six young pigs (weights = 10 to 12 kg) within 30 s on 10 suc- using guinea pigs, rats, and miniature swine in this study. cessive days. The opposite side served as the control field, and Undiluted castor oil (0.1 g) was applied daily to dorsal skin was massaged only. The animals were killed on day 11. A tissue (clipped free of hair) in the mid-lumbar region of six male Hart- section (1 × 1 cm) was excised from the center of the test fields ley guinea pigs (weights = 350 to 500 g) and six male Wistar rats and prepared for microscopic examination. Five to eight sec- (weights = 250 to 350 g). The period of testing, the frequency of tions of 7 µm thickness per test area were used for evaluation. application, and the method of evaluation of skin reactions were

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CASTOR OIL 57 the same as those in the preceding test involving albino angora to 5%), and lactic acid (1% to 5%). The diluted test substance rabbits. Castor oil was mildly irritating to the skin of guinea pigs (concentration after dilution not stated) was applied to clipped and rats. skin on two areas of the back of each animal; one of the areas The skin irritation potential of undiluted castor oil was eval- was abraded. The test sites were covered with occlusive patches uated using six miniature swine of the Pitman-Moore strain (secured with adhesive tape) and the entire trunk was wrapped (1 month old). The test substance (0.05 g) was introduced under a in a rubber sleeve. The patches remained in place for 24 h and 15-mm-diameter patch that was secured with adhesive tape. The reactions were scored at 24 and 72 h post application as fol- entire trunk was wrapped with rubberized cloth during the 48-h lows according the Draize scale: 0 (no erythema) to 4 (severe exposure period. Reactions were scored after patch removal. The erythema [beet redness] to slight eschar formation [injuries in grading scale was the same as that used in the experiment in- depth] and 0 (no edema) to 4 (severe edema, raised more than 1 volving albino angora rabbits that is summarized above. Castor mm and extending beyond area of exposure). oil was not irritating to the skin of miniature swine (Motoyoshi At 24 h post application, well-defined erythema was observed et al. 1979). at the abraded test sites of all six rabbits and slight erythema was observed at the intact sites of four rabbits. Well-defined erythema Ricinoleic Acid was observed at the intact sites of two rabbits during the 24-h In a study by Vieira et al. (2000), 8 to 10 male Swiss mice re- reading. At 72 h post application, five rabbits had mild erythema ceived a single topical application of Ricinoleic Acid (in peanut at the abraded site and two rabbits had the same reaction at the oil, 10 mg/mouse) on the ventral surface of the paw. Neither intact site. Using the data for abraded and intact test sites, a total erythema nor edema of the paw was observed. primary irritation index of 1.1 was calculated (International Bio- Vieira et al. (2001) presented results on the skin irritation Research, Inc. 1977a). potential of Ricinoleic Acid, using groups of six male albino Dunkin-Hartley guinea pigs, in a study evaluating pro- and anti-inflammatory effects of Ricinoleic Acid (concentration not Effect on Intestinal Membranes stated). Ricinoleic Acid (0.1 ml, in peanut oil) was administered Ricinoleic Acid topically to the entire eyelid surface. The thickness of the eyelid In a study by Morehouse et al. (1986), a single 0.1 ml dose of was measured in mm using ophthalmic microcallipers. Topi- Ricinoleic Acid (100 mg/ml) was administered intragastrically cal treatment with Ricinoleic Acid (10, 30, or 100 mg/guinea to fasted, specific pathogen-free mice (strain CD-1, number not pig) caused eyelid reddening and edema. A moderate, dose- stated). This dosage of Ricinoleic Acid was determined, on a dependent eyelid edema was reported as follows: 0.12 ± 0.05 weight basis, to be approximately the dosage that is used ther- mm (10 mg Ricinoleic Acid), 0.18 ± 0.02 mg (30 mg), and 0.23 apeutically in humans. Groups of mice were killed at various ± 0.1 mm (100 mg). Maximal edema was achieved at 2 h post- intervals, and light microscopy and transmission and scanning application. The application of Ricinoleic acid vehicle did not electron microscopy were used to identify structural alterations. cause any appreciable edema. At 2 h post dosing, the duodenal villi were markedly shortened when compared to control duodenal villi. This erosion of the Cetyl Ricinoleate villi throughout the duodenum caused massive exfoliation of In a skin irritation study conducted by the Laboratoire de columnar and goblet cells, filling the lumen with cellular debris Recherche et d’Experimentation (1994), Cetyl Ricinoleate (test and mucus. Disruption of the mucosal barrier resulted in con- concentration not stated) was evaluated using three male New tinuity between the intestinal lumen and lamina priopria of the Zealand albino rabbits. Erythema and edema were observed in villi, with the loss of formed blood elements and lamina propria three and two rabbits, respectively, up to 72 h post application. constituents into the intestinal lumen. The mucosal damage was At 72 h, one rabbit had an erythema score of 2 and the remaining much more localized at4hpost dosing, and the erosion of the two had an erythema score of 1. Edema (score = 1) was observed villi had been largely repaired. Repair was complete at 6 h post in two rabbits at 72 h. All reactions were totally reversible at 7 dosing. days post application. Cetyl Ricinoleate was a nonirritant in this study. Sodium Ricinoleate Gaginella et al. (1977a) studied the morphology of the colon Zinc Ricinoleate after perfusion of the organ with Sodium Ricinoleate solution, In a study by International Bio-Research, Inc. (1977a), using 25 male New Zealand white rabbits. A Foley catheter the skin irritation potential of 10% Griillocin HY 77 (now was inserted and the colon was perfused with control solution TEGODEO HY 77) in soybean oil was evaluated using six (1.25 ml per minute) for 20 min. The colon was then flushed New Zealand albino rabbits (weight range = 2.4 to 2.6 kg). and perfused with the test solutions. Ten rabbits were perfused TEGODEO HY 77 (trade name mixture for Zinc Ricinoleate) with 10 mM Sodium Ricinoleate and the remaining three groups has the following composition: Zinc Ricinoleate (more than of five were perfused with 2.5, 5.0, and 7.5 mM, respectively. 50%), triethanolamine (10% to 25%), dipropylene glycol (1% The animals were killed and full thickness samples of colon

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58 COSMETIC INGREDIENT REVIEW were obtained and prepared for light or scanning electron mi- Castor oil was applied to the external ear five times weekly (one croscopy. In additional experiments, tissue was obtained from application per day). The actual amount applied ranged from 5 animals that were not perfused and those perfused with control to 10 mg/cm2. The animals were killed after the 14th application buffer (contained PEG 400) for 60 or 160 min. and several full-thickness sections of the test site and contralat- Using scanning electron microscopy, mucous membranes of eral untreated ear were prepared for microscopic examination. colons perfused with control buffer for 60 min and unperfused Comedogenic activity was based on the degree of follicular hy- colons appeared normal. After1hofperfusion with 10 mM perkeratosis and other morphologic changes in the majority of Sodium Ricinoleate, patchy loss of epithelial cells was apparent. pilosebaceous units, when compared to control sections. Come- The loss of epithelial cells was confirmed using light microscopy. dogenicity was graded according to the following scale: 0 (neg- A dose response-relationship for mucosal damage was noted. ative: pilosebaceous unit normal in appearance when compared The most severe damage resulted from perfusion with 10 mM to control [untreated] ear section) to 5 (severe: widely dilated Sodium Ricinoleate. Lesser degrees of change and lesser areas follicles, filled with packed keratin; follicular epithelial hyper- of mucosal surface affected were associated with lower concen- plasia causing partial or total involution of sebaceous glands and trations. Small amounts of DNA (due to loss of DNA into the ducts; possible inflammatory changes). Comedogenicity scores lumen) accumulated in the control buffer, and perfusion with 2.5 for castor oil were 0 to 1, corresponding to a slight increase in mM Sodium Ricinoleate did not cause a significant increase in keratin content within the follicle, with essentially no change in these amounts. However, DNA accumulation increased signif- follicular epithelium. icantly after perfusion with 5.0, 7.5, and 10 mM Sodium Rici- Fulton (1989) evaluated the comedogenicity of castor oil nolete, and a plateau was reached with 7.5 mM solutions. (mixed in propylene glycol at 9:1 dilution; test concentration The control perfusate that was used contained low (PEG = 10%) using three New Zealand albino rabbits. The test sub- 400) and high-molecular-weight (PEG 4000) polyethylene gly- stance (1 ml) was applied onto the entire inner surface of one ear cols. The absorption of PEG 400 increased progressively with 5 days per week for 2 weeks. Untreated ears served as controls. increases in the concentration of Sodium Ricinoleate (above Follicular keratosis was evaluated both macroscopically (visu- 2.5 mM) perfused. Sodium Ricinoleate (2.5 mM) did not in- ally) and microscopically, using a micrometer to measure the crease absorption of the individual polymers that comprise PEG width of the follicular keratosis. Follicular keratosis was eval- 400 (molecular weight 242 to 594). The absorption of lower uated according to the following scale: 0 or 1 (no significant molecular weight polymers (242 to 462) increased significantly increase in follicular keratosis) to 4 or 5 (an extensive increase (p <.005) in the presence of 5, 7.5, and 10 mM Sodium Ri- in follicular keratosis). The following scale was used to evaluate cinoleate. For higher molecular weight polymers (506 to 594), skin irritation that resulted from repeated application of the test increased absorption in the presence of 5 mM Sodium Rici- substance: 0 (no irritation) to 5 (epidermal necrosis and slough). noleate was not observed. However, significantly increased ab- A follicular keratosis score of 1 and an irritation score of 0 was sorption (p <.05) of the higher molecular weight polymers reported for castor oil. was observed during perfusion with 7.5 and 10 mM Sodium Ricinoleate (Gaginella et al. 1977a). Skin Sensitization Zinc Ricinoleate Comedogenicity International Bio-Research, Inc (1977b) conducted a skin Castor Oil sensitization study on Grillocin HY 77 (now TEGODEO HY Fulton et al. (1976) evaluated the comedogenicity of cos- 77) using 30 white guinea pigs (average weight = 300 g). metics and cosmetic ingredients. Castor oil was applied to the TEGODEO HY 77 (trade name mixture for Zinc Ricinoleate) base of the internal right ear of each of three white albino rab- has the following composition: Zinc Ricinoleate (more than bits on Monday through Friday for 2 weeks. The left ear served 50%), triethanolamine (10% to 25%), dipropylene glycol (1% as the control. Comedogenicity was clinically evaluated on the to 5%), and lactic acid (1% to 5%) (Degussa 2001). Test and following Monday using the scale: 0 (no increase in visible hy- control groups consisted of 20 and 10 guinea pigs, respectively. perkeratosis) to 5 (severe lesions, such as those seen following A closed patch containing the trade mixture (0.5 ml, undiluted) the application of coal tar). Six-millimeter punch biopsies (right was placed on the left shoulder, clipped free of hair, of each ear) were obtained for microscopic examination, and comedo- animal and remained in place for 6 h. Treatments were repeated genicity was evaluated according to the same scale. A score of once weekly for 3 weeks. 1 (increase in visible hyperkeratosis extending to the possible At two weeks after the last exposure, test and control guinea presence of comedones) was reported for castor oil, which was pigs were challenged (same procedure, right side) with dupli- considered nonsignificant because repeated testing in different cate closed patches. At 24 h post removal of the patches, the test rabbits often failed to produce identical effects. site on each animal was treated with a depilatory. Sites were In a study by Morris and Kwan (1983), the comedogenicity then rinsed and reactions were graded 2, 24, and 48 h later of castor oil was evaluated using three New Zealand rabbits. according to the method of Draize. Clinical observations were

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CASTOR OIL 59 normal throughout the study. Erythema and eschar formation or Triton X-100 extracts of Castor Wax, but a positive response were graded according to the following scale: 0 (no erythema) (positive PCA titer of 40, mean of three determinations) resulted to 4 (severe erythema [beet redness] to slight eschar formation from the reaction of the antiserum with the SLS extract of Castor [injuries in depth]). The following scale for edema formation Wax. was used: 0 (no edema) to 4 (severe edema [raised more than The radioallergosorbent test (RAST) inhibition assay was 1mmand extending beyond area of exposure]). A score of 0 used for further analysis of allergens in the extracts of Castor (erythema and edema) was reported for each test and control Wax. This assay was capable of detecting up to 7.5 µg castor animal during each of the three grading periods. Grillocyn HY allergen. Patient serum (100 µl) and Castor Wax extract (100 µl) 77 did not produce a positive reaction in any of the test animals were incubated with antigen-coated discs. Incubation was fol- (International Bio-Research, Inc. 1977b). lowed by the addition of radioiodinated anti-IgE and another incubation period. Normal serum incubated with antigen coated discs and test serum incubated with discs coated with a non– Allergenicity cross-reacting antigen served as controls. Assay results were Hydrogenated Castor Oil expressed as percent inhibition of the patients’ RAS test. Nei- Lehrer et al. (1980) investigated the presence of castor bean ther the PBS nor Triton X-100 extract of Castor Wax had an allergens in Castor Wax (Hydrogenated Castor Oil) using both inhibitory effect on the RAS test, whereas the urea-NaCl extract in vitro and in vivo analyses. The experiments in this study in- induced a small degree (13.4%) of inhibition. However, the SLS volving human subjects are summarized under the subsection Castor Wax extract induced significant inhibition (40.1%) of the Allergenicity in the Clinical Assessment of Safety section of RAS reaction. this report. The following solvents were used to prepare aque- Based on the nonspecific reaction of the SLS Castor Wax ous extracts of Castor Wax: phosphate buffered saline (PBS), extract with rabbit serum proteins in the first experiment of this urea-NaCl, Triton X-100, or sodium lauryl sulfate (SLS). The study, the authors examined the possibility that this extract may aqueous extracts prepared were analyzed for total organic mat- inhibit the RAST nonspecifically using a heterologous RAST ter and protein (expressed as percent of total organic matter or inhibition assay. An SLS or Triton X-100 extract of Castor Wax dry weight). Results from assays for protein in the aqueous ex- was incubated with reaginic antisera from a patient who was tracts of Castor Wax were as follows: 100% protein (urea-NaCl sensitive to peanuts, and then assayed in the peanut RAST. The extract), 17.0% (SLS extract), 12.9% (PBS extract), and 0.34% SLS extract of Castor Wax inhibited the peanut RAST by 37%, (Triton X-100 extract) protein. suggesting that the results for the RAST inhibition assay in the In a qualitative precipitation analysis, Castor Wax aqueous preceding paragraph may be due entirely or, in part, to a non- extracts were analyzed for castor bean antigens using an im- specific inhibition of the RAST reaction, rather than indicating munodiffusion procedure involving rabbit antiserum (50 µlof that the SLS extract of Castor Wax contains specific castor al- antigen or antiserum in each well). The antiserum was produced lergens. in rabbits that had been immunized with extracts of castor bean The results of these experiments (both in vitro and in vivo) in Freund’s complete adjuvant. Of the aqueous extracts tested, indicate that allergens are present at low concentrations in Castor only the SLS Castor Wax extract reacted with the rabbit anti- Wax, because the SLS Castor Wax extract induced positive PCA sera. No reactivity with the antiserum was observed when the reactions in mice and a positive direct RAS test for reaginic SLS solution alone was tested. Because the reaction was unusu- castor allergens (Lehrer et al. 1980). ally strong, suggesting a false-positive reaction, the extract was tested with normal rabbit serum. However, the same precipitin line was observed. It was concluded that the reaction of the SLS Castor Wax extract with rabbit serum proteins was probably a NEUROTOXICITY nonspecific reaction. Sodium Ricinoleate The aqueous extracts of Castor Wax were also analyzed for Foxetal. (1983) applied Sodium Ricinoleate (0.01% to 1.0%) allergens by determining their reactivity (48-h passive cutaneous to the serosal surface of the rat (anesthetized male Sprague- anaphylaxis [PCA] reaction) with mouse anticastor reaginic (im- Dawley rats, weights = 200 to 225 g) jejunum every 5 min munoglobulin E [IgE]) antibody. CFW female mice were sen- for 0.5 h. At 30 days post application, treated and untreated sitized with reaginic antibody that was produced in BDF mice jejunal segments were removed and examined microscopically. that had been immunized with heated castor bean extracts and At a concentration of 0.1%, Sodium Ricinoleate significantly Bordetella pertussis adjuvant. The mice were challenged 48 h reduced (p <.02) the number of ganglion cells in the myenteric later by intravenous injection of extracts of Castor Wax in 0.5% plexus. The highest test concentration (1% Sodium Ricinoleate) Evans blue solution and killed at 30 min post injection. Non- significantly reduced the number of cells in the myenteric plexus sensitized mice challenged with Castor Wax extracts served as (p <.01) and the submucosal plexus (p <.02). The lowest test controls. The reactions in control mice were always classified concentration (0.01%) did not significantly reduce the number as negative. The antiserum did not react with PBS, urea-NaCl, of ganglion cells in the myenteric or submucosal plexus.

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60 COSMETIC INGREDIENT REVIEW

Renal Toxicity mg/kg cyclosporine dose; administered to test group of seven) in Castor Oil aregion of the upper right abdomen. Dosing was followed by five Langer et al. (1968) evaluated effects of castor oil on the rat 20-min clearance periods. Compared to the mineral oil control kidney, following intraluminal injection, using five male white group (six rats), a significant decrease (25% decrease; p <.05) albino rats (weights = 180 to 220 g). The kidney was exposed in renal blood flow was noted in rats (group of five) dosed i.p. and proximal tubuli were punctured (using Leitz stereomicro- with castor oil. Renal vascular resistance was also significantly scope) and filled with castor oil. At 15 to 60 min, the proximal higher (p <.05). No significant changes in inulin clearance tubular segments (filled with oil) were fixed in situ. The kidney were reported. A significant decrease in blood pressure was wasexcised and prepared for both light and electron microscopic noted in all groups; however, blood pressure remained within examination. The injection of castor oil resulted in dilatation of the physiologic and autoregulatory range (Racusen et al. 1986). the proximal convoluted tubular lumen and compression of the brush border of proximal tubular cells, although a cessation of the normal pinocytosis was induced in the presence of tubular GENOTOXICITY fluid. A toxic effect of castor oil on the tubular epithelium was Castor Oil not observed. Zeiger et al. (1988) evaluated the mutagenicity of castor oil In a study by Racusen et al. (1986) evaluating the nephrotoxi- (United States Pharmacopeia purity grade; in dimethylsulfox- city of cyclosporine, castor oil served as the vehicle control. Two ide [DMSO]) in the preincubation assay using Salmonella ty- control groups of eight adult male Munich-Wistar rats (weights phimurium strains TA97, TA98, TA100, TA1535, and TA1537. = 220 to 270 g) were used. Group 1 vehicle controls (vehicle- The assay was conducted, with and without metabolic activation, ischemia group) underwent bilateral renal artery clamping, and according to a modification of the procedure by Haworth et al. were then dosed intraperitoneally (i.p.) daily (4-day period) with (1983). Castor oil was tested at doses up to 10,000 µg/plate avolume of castor oil vehicle that was equivalent to the i.p. dose and classified as nonmutagenic (all strains, with and without of cyclosporine (dose = 60 mg/kg). Group 2 vehicle controls metabolic activation) over the range of doses tested in this as- (vehicle-sham group) underwent sham surgery. The incisions say. were closed prior to daily i.p. dosing with castor oil. On postop- Hachiya (1987) evaluated the mutagenicity of castor oil using erative day 3, feed was withdrawn and urine was collected over a S. typhimurium strains TA97, TA98, TA100, TA102, TA1537 period of 12 h for clearance studies. On postoperative day 4, the and strain WP2/pKM102 of Escherichia coli.Ineach strain, animals were killed and tissues prepared for light and electron castor oil was tested at doses ranging from 100 to 5000 µg/plate microscopy (confined to cortical proximal convoluted tubules). with and without metabolic activation. Results were negative for In separate studies, renal blood flow and the glomerular fil- all strains tested, both with and without metabolic activation. tration rate were determined. Two groups of eight saline control Cytotoxicity (at doses of 100 to 5000 µg/plate) was noted only groups (saline-ischemia and saline-sham groups, respectively) in strain WP2/pKM102, with and without metabolic activation. were treated according to the same procedure. NTP (1992) reported a study in which groups of male and Tubular vacuolization was a prominent feature in vehicle- female B6C3F1 mice received diets containing 0.62%, 1.25%, ischemia control rats, and was observed to a minor degree in the 2.5%, 5.0%, and 10% castor oil (in DMSO) for 13 weeks. Blood vehicle-sham control rats. The clear vacuoles observed using samples were obtained at the end of the 13-week study. There light microscopy probably corresponded to two ultrastructural wasnoevidence of induction of micronuclei in peripheral ery- changes, dilated endoplasmic reticulum and lipid droplets. With throcytes. the exception of the saline-sham group, dilated endoplasmic NTP (1992) also evaluated the induction of sister chromatid reticulum was observed in all control animals. However, lipid changes by castor oil (in DMSO) in Chinese hamster ovary cells. droplets were not observed in any of the control animals. The The cells were incubated with castor oil at concentrations up to presence of eosinophilic cytoplasmic inclusions was reported for 5000 µg/ml with and without metabolic activation. Mitomycin- two of eight vehicle-ischemia rats and four of eight vehicle-sham C and cyclophosphamide served as positive controls without rats, but not for saline-control rats. Tubular necrosis (confined and with metabolic activation, respectively. DMSO served as entirely to outer medullary pars recta) was observed in half of the the solvent control. Castor oil did not induce sister chromatid vehicle-ischemia rats and in all saline-ischemia rats, but was not exchanges either with or without metabolic activation. observed in saline-sham or vehicle-sham control groups. Tubular regeneration was noted in vehicle-ischemia and saline-ischemia Sodium Ricinoleate control rats. In a study by Haworth et al. (1983), the mutagenicity of The renal clearance study involved two control groups that Sodium Ricinoleate was evaluated according to the preincu- were dosed with castor oil (five rats) or mineral oil (six rats). Two bation procedure of the Salmonella assay (Ames et al. 1975) 20-min inulin clearance determinations were made, and the vehi- as described by Yahagi et al. (1975). Sodium Ricinoleate (in cle was then administered i.p. (at dose volume comparable to 60 distilled water) was tested at concentrations up to 33 µg/plate

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CASTOR OIL 61 at one testing laboratory, and up to 333.3 µg/plate at another Tumor Promotion Activity laboratory. The following S. typhimurium strains were used Castor Oil, Ricinoleic Acid, and Glyceryl Ricinoleate with and without metabolic activation: TA98, TA100, TA1535, Shubik (1950) evaluated the tumor promotion activity of cas- and TA1537. Distilled water served as the vehicle control. The tor oil, Ricinoleic Acid, Glyceryl Ricinoleate and the following positive controls were as follows: 2-aminoanthracene, 4-nitro- other chemicals using groups of mice (strain and weights not o-phenylenediamine, sodium azide, and 9-aminoacridine. Re- stated): 20% turpentine in liquid paraffin, 0.3% acridine in liq- sults for Sodium Ricinoleate were negative with and without uid paraffin, 0.5% fluorene in liquid paraffin, 1% phenanthrene metabolic activation. in liquid paraffin, silver nitrate (10% aqueous), and oleic acid. Castor oil, Ricinoleic Acid, Glyceryl Ricinoleate, and oleic acid were not diluted prior to testing. DNA Binding of Reaction Product Initially, the nine substances were tested using groups of three Methyl Ricinoleate mice (one group per substance). The test substances were ap- Frankel et al. (1987) evaluated the fluorescence formed by the plied to a small area of skin in the interscapular region (clipped interaction of DNA with lipid oxidation products. The keto ester, free of hair). Four applications per mouse, spaced over a period methyl ketoricinoleate, was prepared from Methyl Ricinoleate of 2 weeks, were made. At the end of the application period, by allylic oxidation with a CrO3-pyridine complex in methylene the animals were killed and tissues were obtained for micro- chloride. Results from this assay indicated that methyl ketori- scopic examination. The following substances induced slight cinoleate (relative concentration = 1 mM/3 ml) produced very epidermal hyperplasia: castor oil, Ricinoleic Acid, Glyceryl Ri- little fluorescence (mean = 29 ± 2 units) with DNA in the pres- cinoleate, oleic acid, fluorene, and phenanthrene. Turpentine in- ence of ferric chloride–ascorbic acid. The mean value for methyl duced minimal histologic change, and acridine induced marked linolenate hydroperoxides (used as reference) was 362 ± 9 units. epidermal hyperplasia. Silver nitrate induced hyperplasia most Mean values were given as the fluorescence intensity of DNA marked in the hair follicles. after 72 h (excitation: 315 nm; emission: 420 nm). In the tumor promotion experiment (groups of 10 mice; strain and weights not stated), each mouse received a single application of 9,10-dimethyl-1,2-benzanthracene in liquid paraffin. The nine test substances (one per group) were then applied semiweekly CARCINOGENICITY for 20 weeks. For all groups of mice tested, no tumors were Ricinoleic Acid recorded. Castor oil, Ricinoleic Acid, and Glyceryl Ricinoleate, In a study by Boyland et al. (1966), 2% Ricinoleic Acid, as well as the other test substances that were evaluated, were not in gum tragacanth, was injected intravaginally into each of 20 tumor promoters in this study (Shubik 1950). BALB/c female mice (5 to 6 weeks old). Injections were made twice per week, and each animal received a total of 100 injections. According to the same dosing schedule, each of 20 Ricinoleic Acid positive control and 30 vehicle control mice received 62 injec- Bull et al. (1988) investigated the effect of Ricinoleic Acid tions of 0.3% dimethylbenz(a)anthracene and 100 injections of (unoxidized) on DNA synthesis and the induction of ornithine gum tragacanth, respectively. An additional 30 mice served as decarboxylase activity. The authors noted that unsaturated fatty the untreated control group. acids are readily oxidized to a variety of biologically active None of the 20 mice injected with Ricinoleic Acid had neo- derivatives, and have suggested that autoxidation products of plasms or hyperplastic lesions of the corpus uteri, cervix uteri, polyunsaturated fatty acids may also play a role in the enhance- vagina, or perineal skin. However, benign lung adenomas were ment of tumorigenesis. They noted that this suggestion is based observed in ten of the 13 mice dosed with Ricinoleic Acid and on published results (Bull et al. 1984) indicating that hydroper- in 6 of the 24 vehicle control mice that were killed after the 14th oxy and hydroxy derivatives of arachidonic acid and linoleic month of dosing. Benign lung adenomas were also observed acids, the primary autoxidation products of major dietary unsat- in 9 of the 20 untreated control mice that were killed after the urated fatty acids, induce ornithine decarboxylase (ODC) and 14th month. The difference in benign adenoma incidence be- stimulate DNA synthesis (two components of mitogenesis) in tween Ricinoleic Acid–treated and untreated control mice was colonic mucosa in vivo. not statistically significant (p > .1). Additionally, compared Male Sprague-Dawley rats (weights = 100 to 125 g) fed to controls, there was no tendency for Ricinoleic Acid–treated the following semisynthetic diet (weight percentages indicated) mice with adenomas to have a higher average number of tumor were starved for 24 h prior to intrarectal (i.r.) instillation of Rici- nodules per mouse. Malignant tumors of the vagina and perineal noleic Acid (vehicle: 5% ethanol in 50 mM NaHCO3,pH8.0): skin were observed in 15 of the 20 (75%) positive control mice. casein (20%), DL-methionine (0.2%), salt mix (3.6%), vitamin The authors stated that the results of this study do not provide mix (2%), beef fat (5%), dextrose (45.2%), cellulose (5%), and any definite evidence of the carcinogenicity of Ricinoleic Acid cornstarch (19%). Two experiments were performed using Ri- (Boyland et al. 1966). cinoleic Acid test concentrations of 5 and 10 mM. Test groups

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62 COSMETIC INGREDIENT REVIEW consisted of three to ﬁve rats per dose level. The vehicle control from mice inoculated with ARS cancer cells (with well-grown group (three rats per experiment) was dosed with 5% ethanol in ascites) that had been washed with physiological brine in order 50 mM NaHCO3, and positive control animals received 6 mM to collect the cancer cells. The physiological brine was diluted deoxycholate. Data were presented as an average of the two ex- and injected (0.2 ml) into the abdominal cavity of each mouse. periments. The measurement of DNA synthesis in the colon was On the day after injection of the cancer cells, each of nine mice based on the incorporation of [3H]dThd 12 h after i.r. instilla- was injected with a polyphase liposome that was produced from tion of Ricinoleic Acid. ODC activity was determined in colonic castor oil extract (dose = 200 mg/kg) daily for 7 days. At the mucosa that was harvested3hafter i.r. instillation of Ricinoleic end of the treatment period, the animals were monitored (for Acid. survival) for an additional 35 days. Ricinoleic Acid was inactive as a stimulator of DNA synthesis Study results indicate that the castor oil extract had a strong and inducer of ODC activity in the rat colon. Concentrations suppressive effect on S180body tumors in mice. Its suppressive up to 10 mM resulted in insigniﬁcant increases in [3H]dThd effect on ARS ascites cancer was also very strong, with 64% of incorporation and did not increase the level of ODC activity the ascites tumors in mice being completely cured. Compared to above control values (Bull et al. 1988). controls, the life extension rate was more than 136% (p <.0012) (Xie et al. 1992).

Anticancer Activity Extract of Castor Oil REPRODUCTIVE AND DEVELOPMENTAL TOXICITY Xie et al. (1992) evaluated the anticancer/antitumor activity Castor Oil of an extract of castor oil using male Kunming mice (weights NTP (1992) (see earlier Subchronic Oral Toxicity section) = 18 to 22 g) and mice bearing S180 ARS ascitic fluid tumor fed groups of rats and mice diets containing 0.62%, 1.25%, cells. Castor oil was saponified and acidified to obtain a coarse, 2.5%, 5.0%, and 10% castor oil, respectively, continuously fatty acid mixture that was recrystallized and refined to produce for 13 weeks. A slight decrease in epididymal weight (6% to the active anticancer component. Using Tween-80 (Polysorbate 7%) was observed in mid- and high-dose groups of male rats; 80), an emulsion containing this component was produced. A however, this finding was not dose related. No effects on any polyphase liposome from castor oil extract was also produced. other male reproductive end point (testes weight and epididy- The inhibitory action of the anticancer component on S180 tumor mal sperm motility, density, or testicular spermatid head count) cells was evaluated. Ascites was obtained from mice that had or female reproductive endpoint (estrous cycle length, or time been inoculated with S180 tumor cells (with well-grown ascites) spent in each phase of the cycle) were noted. Microscopically, that had been washed with physiological brine in order to collect there were no treatment-related lesions in any organ or tissue. the cancer cells. The physiological brine was diluted and injected These results indicate that there was little or no evidence of any (2 ml) subcutaneously into the right front axilla of each mouse. reproductive toxicity in rats that was associated with castor oil in On the day after injection of the tumor cells, the mice were the diet. For male and female mice, castor oil in the diet had no randomly divided into groups. adverse effects on any male or female reproductive parameter. The three dilution control groups (8 to 10 mice per group) Litvinova and Fedorchenko (1994) evaluated the influence of were injected with 200, 300, or 400 mg/kg of a physiologi- single doses of different plant oils on the estrous cycle and fer- cal brine solution containing an equal load of Tween-80. The tility using female Wistar rats (number and weights not stated). four polyphase liposome control groups (eight mice per group) Castor oil (0.2 ml, single dose) was injected intramuscularly on were injected with blank liposomes (doses of 300, 300, 400, and the first day after estrus. Study of the estrous cycle was based on 400 mg/kg, respectively). For dilution control and polyphase li- the following: the cytologic picture that was obtained from vagi- posome control groups, injections were made into the abdominal nal smears, the phase structure (proestrus, estrus, metaestrus, and cavity daily for 7 days, after which the mice were killed. Subcu- diestrus) of the cycle, and the frequency of estrus over a 15-day taneous tumors were removed and weighed in order to calculate observation period prior to injection of castor oil (control) and the tumor suppression rate (%). a similar length of time after injection (test). Fertility was stud- Tumor suppression rates for dilution controls were as fol- ied using intact animals (control) and animals that were treated lows: 30.6% (200 mg/kg dose group), 37% (300 mg/kg dose with castor oil. On day 2, after castor oil injection (i.e., during group), and 53.1% (400 mg/kg dose group). The following tu- diestrus), the females were mated with male rats. Control fe- mor suppression rates were reported for the polyphase liposome males were also mated with male rats on day 2. The first day control groups: 40.2% (300 mg/kg dose group), 36.4% (the other of pregnancy was defined by the presence of spermatozoa in 300 mg/kg dose group), 55.7% (400 mg/kg dose group), and vaginal smears. Pregnant females were killed on day 20. The es- 58% (the other 400 mg/kg dose group). trous cycle phases were recorded 3 to 4 times during the 15-day In another experiment, the inhibitory action of the anticancer observation period. component extracted from castor oil on ARS ascites cancer was The injection of castor oil resulted in a reduction in the evaluated according to a similar procedure. Ascites was obtained frequency of estrus by a factor of 1.4 and lengthening of the

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CASTOR OIL 63 interestrous phase by 39.6%. The authors concluded that the treatment, there was no difference in male reproductive perfor- latter finding may be indicative of prolongation of luteolytic mance between ICI-treated and control rats. However, after day processes and an inhibition of folliculogenesis. 100, 100% fertility was maintained in control males, but, by day In female fertility experiments, mating occurred over a pe- 150, fertility had decreased to 25% in ICI-treated males (Oliveira riod of 3.22 ± 0.12 days. The index of fertility was calculated et al. 2001). from the ratio of the number of mated females to the total number of females in the test. The fertility index for intact an- Extract of Ricinus communis Seeds imals (control) was 100%, and pregnancy occurred in 100% The following study is included because castor oil may be of the cases. A decrease in the pregnancy index was reported extracted from the bean of the tropical plant, Ricinus communis for female rats injected intramuscularly with castor oil; how- by a method that involves the use of a solvent. ever, no reliable difference between test and control values was Okwuasaba et al. (1991) evaluated anticonceptive and es- detected. trogenic effects of a methanol extract of Ricinus communis Compared to controls, castor oil also caused a reduction in var.minor seeds (ether-soluble fraction) using the following an- the number of corpora lutea in the ovaries, and this observa- imals (groups of 10): adult albino Wistar rats (weights = 160 to tion was said to have been a consequence of the suppression of 210 g), young albino Wistar rats (weights = 40 to 60 g), imma- folliculogenesis and ovulation. Additionally, the number of im- ture Swiss albino female mice (21 to 23 days old; weights = 10 plantation sites in uteri from female rats injected with castor oil to 13 g), and adult female New Zealand white rabbits (2.5 to 3.0 was decreased when compared to control rats. A decrease in the kg). number of live fetuses in uteri and an increase in pre- and postim- In the antifertility experiment, adult female rats and rabbits plantation deaths was also observed in rats injected with castor with regular estrus cycles were used. The females were mated oil. The authors concluded that castor oil, injected intramus- overnight (3:1 female/male ratio), and the day of mating was cularly, suppressed ovarian folliculogenesis and also had anti- considered day 0. Castor bean extract was injected subcuta- implantation and abortive effects (Litvinova and Fedorchenko neously (s.c. dose of 0.6 or 1.2 g/kg) into female rats once daily 1994). for two days, and mating occurred on the third day. Female Castor oil served as the vehicle control in a study evaluating rabbits were injected intramuscularly with Castor bean extract the effect of long-term treatment with ICI 182,780 (an antie- (dose = 200 mg/kg). Control animals were injected s.c. with strogen) on the rat testis (Oliveira et al. 2001). Two groups of corn oil for two consecutive days. A laparotomy was performed four male, 30-day-old Sprague-Dawley rats served as vehicle to confirm that implantation had occurred. controls. In one control group, castor oil (0.2 ml) was injected Castor bean extract (0.6 or 1.2 g/kg dose) prevented nida- subcutaneously once per week, and the animals were killed 100 tion in rats, and none of the treated rats delivered pups at days after the first injection. The second group was dosed accord- term. The results for rabbits (200 mg/kg dose) paralleled those ing to the same procedure; animals were killed 150 days after the for rats. Nidation was prevented, and the extract protected first injection. The experiment was extended to 150 days (group the rabbits against pregnancy for over three gestation periods 2 rats), based on results for group 1 ICI-treated and control ani- (i.e., 110 to 120 days). The normal gestation period for rab- mals on day 100. bits is 28 to 30 days. Effects induced by Castor bean extract Specifically, Group 1 ICI-treated rat testes were significantly were reversible in rats and rabbits. No fetal abnormalities were heavier (35% heavier, p <.05) than those of the control group, observed. and a significant increase in diameter and luminal areas of the In the study to determine estrogenic activity, the following semineferous tubules was observed. Additionally, in group 1 endpoints were used to estimate the estrogenicity of the Castor ICI-treated and control rats, spermatogenesis appeared normal bean extract: uterine weight ratio, degree of vaginal cornifica- and there was no evidence of degeneration or sloughing of germ tion, and quantal vaginal opening. In one experiment, groups cells. of 10 Swiss albino female mice (21 to 23 days old; weights On day 150, a 33% decrease (compared to day 100 results for = 10 to 13 g) were injected s.c. with 1.0 or 4.0 mg/kg Castor ICI-treated rats; p = .065) in mean testicular weight was noted bean extract (in corn oil, 0.2 ml) daily for 4 days. In a similar in ICI-treated rats. Seventy-five percent of the testes weighed experiment, bilateral ovariectomy was performed on young, im- less than the mean control value of 1.71 g. Atrophy of the sem- mature rats (groups of five), and the wound was sutured. After ineferous tubules was observed in all but one of the testes of a 15-day recovery period, the groups of rats were injected s.c. ICI-treated rats. Additionally, the rete testis and efferent ductule with 0.6 or 1.2 g/kg Castor bean extract (in corn oil, 0.2 ml) lumens in all ICI-treated rats were greatly dilated, compared to daily for 4 days. In both experiments, suitable controls (injected those of controls. s.c. with corn oil only) were maintained. Additionally, estradiol- During the treatment period, mating studies were performed 17β (reference hormone; dose = 10 µg/kg) was administered to compare the fertility of ICI-treated and control rats on days to rats (group of 10) and mice (group of 10). Mice and rats were 45, 73, 100, 125, and 150. Males were housed individually with necropsied 24 h after the last injection. Body and uterine wet two adult females for a 15-day period. Through 100 days of weights were recorded.

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Results for bilaterally ovariectomized rats indicated that 3,6-epoxydecanedioic acid; and 3,6-epoxydodecanedioic acid. doses of 0.6 and 1.2 g/kg induced increases in relative uter- These three Ricinoleic Acid metabolites were also detected in ine weight of 116.7 ± 6.5 (p <.01) and 211.6 ± 8.2 mg/100 g the urine of rats as described earlier. body weight (p <.001), respectively, compared to a control value of 83.3 mg/100 g body weight. Compared to rats, Castor bean extract (1.0 and 4.0 mg/kg) induced a qualitatively similar Induction of Labor effect in immature mice. In rats and mice, the effect on relative Castor Oil uterine weight appeared to have been dose-dependent. Estradiol- Davis (1984) investigated the use of castor oil to stimulate 17β also increased uterine weight in rats and mice. The relative labor using 196 patients with premature rupture of membranes uterine weights (mg/100 g body weight) for rats and mice dosed (PROM), at least 4 h in duration, who were between 37 and with estradiol-17β were reported as follows: 309.2 ± 11.4 (rats) 42 weeks of gestation. Patients were identified by reviewing and 517.5 ± 12.3 (mice). charts of all patients who were admitted at an out-of-hospital Cornification of the vaginal epithelium and quantal open- birthing center from 1976 through 1981. Of the 196 patients, 107 ing of the vagina were noted in immature mice and bilaterally (mean age = 28.6 years) were dosed orally with castor oil (2 oz) ovariectomized rats (both dose groups for each). A dose-related and 89 (mean age = 27.6 years) were not. Castor oil was adminis- decrease in the number of leukocytes and increases in cornified tered only to PROM patients who had a latency period of at least 4 and epithelial cell counts were also reported for both species. h. All patients were observed for labor onset for 24 h after the on- No significant changes in body weight were observed. set of PROM. If labor did not occur at this time, the patients were The authors concluded that the ether-soluble portion of the transferred to a hospital for oxytocin stimulation. The longest methanol extract of Ricinus communis var. minor seeds pos- interval between rupture of the membranes and delivery was sesses anti-implantation, anticonceptive, and estrogenic activity 48 h. in rats and mice when administered subcutaneously (Okwuasaba Of the 107 patients dosed with castor oil, 80 (75%) had labor et al. 1991). onset. Spontaneous labor occurred in 52 (58%) of the 89 control patients. This difference between patients dosed with castor oil and controls was statistically significant (p <.05). The interval CLINICAL ASSESSMENT OF SAFETY between castor oil administration and the onset of labor ranged from 1 to 13 h (mean = 4 h). Metabolism and Excretion Labor outcomes were also evaluated for type of delivery, in- Castor Oil cidence of oxytocin stimulation, and infant well-being. The need Watson et al. (1963) studied the absorption and excretion for cesarean sections was nearly three times greater in the control of castor oil in five subjects (ages and weights not stated). The group (15.7% incidence) than in patients dosed with castor oil composition of the castor oil was as follows: palmitic acid (1%), (5.6% incidence). This difference was found to be statistically palmitoleic acid (0.1%), stearic acid (1%), oleic acid (3.2%), significant (p <.01). Additionally, the group dosed with castor linoleic acid (4.7%), and Ricinoleic Acid (90%). The castor oil oil did not require oxytocin stimulation as frequently (36% in- (of medicinal purity) contained a trace amount of oil labelled cidence) when compared to the control group (43% incidence), with 131I, and the doses administered ranged from 4 to 60 g and the difference was not statistically significant. (approximately 6 µCi of radioactivity per dose). Castor oil was Infant outcomes in the two groups were studied by observing administered to the five hypertensive patients on the day before a Apgar scores at 1 and 5 min, and looking for any evidence of scheduled intravenous pyelogram; thereafter, only a light meal meconium staining. Two infants with Apgar scores of <7 were of coffee and toast was allowed. Stool collections were made reported for both groups. At the time of membrane rupture, the during the first 24 h after dosing and during the subsequent 72 h. presence of meconium was very low in both groups. Meconium Urine was collected in 24-h samples. Small doses of oil were staining was not observed after dosing with castor oil. It is also administered to the three normal volunteers, and free diet was important to note that there were no maternal deaths and no sig- allowed. Stools were collected at 2 and 3 days after dosing. nificant maternal morbidity. The authors concluded that castor Fecal recovery of 131I (%) ranged from 11.4% (for 10 g dose oil can be used safely and effectively to stimulate labor (Davis of castor oil) to 86.0% (for 44.4 g dose of castor oil). The authors 1984). concluded that castor oil can be absorbed by human subjects, that In a study by Mitri et al. (1987), the amniotic fluid was absorption is inversely related to the administered dose, and, at examined at the time of rupture of membranes in 478 of the small doses (4 g), that absorption is virtually complete (Watson 498 women in labor. The amniotic fluid was meconium-stained et al. 1963). in 174 subjects (mean age = 28.5 ± 0.65 years) and clear in Hagenfeldt et al. (1986) administered castor oil (10 to 15 ml) the remaining 304 subjects (mean age = 25.9 ± 0.37 years). orally to three healthy subjects. Urine was collected between Cesarean section, low Apgar score, and the recent ingestion 2 and 8 h post dosing. The following three epoxydicarboxylic of castor oil were significantly more common in subjects with acids were excreted in the urine: 3,6-epoxyoctanedioic acid; meconium-stained amniotic fluid. The authors concluded that

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CASTOR OIL 65 the passage of fetal meconium was predictive of poor labor out- Skin Irritation/Dermal Toxicity come in terms of Apgar scores and the need for cesarean section. Castor Oil It was recommended that women who take castor oil in late preg- Motoyoshi et al. (1979) evaluated the skin irritation poten- nancy should be identified as a high-risk group for the passage tial of undiluted castor oil using 50 adult male volunteers (ages of fetal meconium. not stated). Individuals with known allergic reactions were ex- In a case report by Steingrub et al. (1988), a 33-year-old cluded. Patches containing 0.05 g of the test material were ap- pregnant female (at week 40 of gestation) ingested castor oil to plied to skin of the back and remained for 48 h. At the end of induce labor. Within 60 min of ingestion, cardiopulmonary arrest the application period, test sites were swabbed with dry gauze occurred and was reportedly due to amniotic fluid embolism. to remove any residual test material and reactions were scored Garry et al. (2000) evaluated the use of castor oil to induce 30 minutes later according to the following scale: − (negative labor in 52 pregnant women (mean age = 24.8 ± 6.7 years) reading) to ++++(bullous reaction). Castor oil was mildly at Saint Mary’s Hospital in Brooklyn, New York. The untreated irritating to the skin of human subjects. control group consisted of 48 pregnant women (mean age = 24.4 In a study by Meyer et al. (1976), undiluted castor oil was ± 4.9 years). The two groups of women did not differ in mater- applied to test fields delineated on the right thigh of each of three nal age, parity, or gestational age. Castor oil was administered male subjects (22 to 31 years old). The control field was located as a 60-ml dose in orange or apple juice, and its use was deemed on the opposite side. According to the test procedure, castor oil successful only if active labor began within 24 h. Labor was was rubbed onto the skin gently (<30 s) on 10 sucessive days. defined as one or more contractions every 5 min, with cervi- The control field was massaged only. A tissue section (1 × 1 cal dilatation of 4 cm or more. Active labor was induced in 30 cm) was excised from the center of each test field and five to (57.7%) of the 52 women dosed with castor oil, compared to 2 eight sections (7 µminthickness) per test area were subjected to of the 48 women in the control group (p <.001). The cesarean microscopic examination. Macroscopic skin changes were not section rate for women dosed with castor oil was 19.2% (10 of observed in either of the three subjects. 52 women), compared to 8.3% (4 of 48 controls) in the untreated At microscopic examination, clear hyperchromasia and an control group. No relationship between dosing with castor oil, increase in the number of cells were observed in the basal cell birth weight, and mode of delivery (p = .66) was found. Ad- layer. Slight widening of the granular cell layer was also noted ditionally, no adverse outcomes for the mother or fetus were (Meyer et al. 1976). identified. The authors concluded that women dosed with castor oil had an increased likelihood of initiation of labor within 24 h, Hydrogenated Castor Oil compared to women who were not dosed with castor oil. In a study by de Groot (1994), members of the Dutch Contact Dermatitis Group and nonmember dermatologists patch tested 1 to 20 patients (ages not stated) either suffering from or suspected Ocular Irritation of suffering from contact allergy to cosmetic products. Details Castor Oil regarding the patch test procedure were not included. Patch test In a study by Secchi et al. (1990), a double-masked clinical results indicated no reactions to 30% Hydrogenated Castor Oil trial was performed using a group of nine patients (ages not in petrolatum. stated) with vernal keratoconjunctivitis. One eye was treated with 2% cyclosporine in castor oil four times daily for 15 days. The other eye was treated with castor oil solution only according Ethyl Ricinoleate to the same procedure. Mild and transient discomfort and minor The skin irritation potential of 20% Ethyl Ricinoleate in epithelial changes were observed in both eyes. petrolatum was evaluated using 32 healthy male volunteers (ages In another clinical study (Gluud et al. 1981), vital staining not stated) in a 48-h closed patch test. Skin irritation was not ob- was used to investigate the eyes of 100 patients (ages not stated) served (RIFM 2000). following the instillation of castor oil and the eyes of 50 patients following the instillation of 0.9% saline. One drop (0.01 ml) Allergenicity of sterile fluorescein–rose bengal (RB) dye mixture was instilled into the lower conjunctival sac and the ocular mucosa Predictive Tests wasexamined using a slit lamp. White light was used to exam- Ethyl Ricinoleate ine RB-stained spots and cobalt blue light was used to examine The skin sensitization potential of 20% Ethyl Ricinoleate in fluorescein-stained spots. The ocular mucosa was divided into petrolatum was evaluated using 32 healthy male volunteers in five regions, and the number of stained spots in each region was the maximization test. The test substance was applied (under counted/estimated and graded. The authors concluded that cas- occlusion) to the same site on the forearms of each subject for tor oil affected the cornea and conjunctiva, i.e, degeneration and five alternate-day 48-h periods. Each test site was pretreated with death of epithelial cells (stained with RB) and continuity breaks 5% aqueous sodium lauryl sulfate (under occlusion) for 24 h. in the epithelium (stained with fluorescein). At the end of a 10- to 14-day nontreatment period, challenge

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66 COSMETIC INGREDIENT REVIEW patches were applied (under occlusion) to new test sites for 48 plied to the skin for 24 and 48 h. Reactions were scored at 24 h. Challenge applications were preceded by 30-min applications and 48 h post removal. of 2% aqueous sodium lauryl sulfate (under occlusion) on the The three patients were highly sensitive to castor allergen in left side. On the right side, the challenge application was not the prick test (positive skin reaction to 1.0 or 0.1 µg/ml castor preceeded by treatment with sodium lauryl sulfate. A fifth site bean extract). The patients also had a positive skin reaction to the (challenge with petrolatum) served as the control. Reactions SLS Castor Wax extract, where the level of sensitivity was 100- classified as either significantly irritating or allergic were not to 100,000-fold less than that induced by castor bean extract. observed (RIFM 2000). None of the seven normal subjects had a delayed reaction to any of the four Castor Wax extracts that were tested. Prick test results (normal subjects) indicated no immediate skin reactivity Provocative Tests to castor bean extract or the SLS Castor Wax extract. Castor Oil Because immediate hypersensitivity to Castor Wax was ob- Fujimoto et al. (1997) conducted a study involving 332 served in the three patients who were sensitive to castor bean, ad- ≈ patients (25 males, 307 females; ages 20 to 70) suspected ditional tests (prick tests) were performed to determine whether of having cosmetic contact dermatitis. The subjects were Castor Wax solubilized in different organic solvents or an un- patch-tested with numerous cosmetic products and cosmetic in- derarm deodorant stick containing Castor Wax would induce gredients. Patch tests (Finn chambers) were applied to the back irritation and elicit a positive immediate hypersensitivity reac- for 48 h, and the test sites were examined at 30 minutes after tion in these three patients. Positive skin reactions to Castor Wax patch removal, 1 day later, and 4 to 5 days after patch removal. (in different solvents or in a deodorant stick) were not detected. None of the 49 patients patch-tested with castor oil had a positive The four Castor Wax extracts and paraffin (control) were reaction. evaluated in patch tests for cell-mediated hypersensitivity. None In a study by Hino et al. (2000), 346 patients (31 males, 315 of the three patients had a delayed reaction to any of the four ≈ females; ages 20 to 70 years) suspected of having cosmetic Castor Wax extracts (test concentration not stated). dermatitis were patch tested with various cosmetic products and The positive skin reactions (immediate skin reactivity) in cosmetic ingredients. Patch tests (Finn chambers) were applied sensitized individuals, together with the positive PCA reactions to the back for 48 h. Test sites were examined according to the in mice and positive direct RAST for reaginic castor allergens schedule in the preceding study. Of the 76 patients patch-tested (summarized earlier in this report) that were induced by SLS with castor oil, one had a positive reaction. This reaction was Castor Wax extract, indicate that allergens are present in Castor observed only during the second reading (i.e., the day after the Waxatlow concentrations. 30-min reading). A direct RAS study was conducted to determine whether the SLS extract of Castor Wax contains specific antigens or non- Hydrogenated Castor Oil specifically inhibits the RAS assay. Discs were coated with cas- Lehrer et al. (1980) evaluated the sensitivity of three human tor bean extract or the SLS Castor Wax extract and a direct RAS subjects (with occupational hypersensitivity to castor allergens) assay was performed using serum from one patient who was sen- to castor bean extract and four Castor Wax extracts (solvent = sitive to castor allergens, a patient who was sensitive to peanuts, SLS, PBS, urea-NaCl, or Triton X-100). Seven normal labora- or pooled human serum (control). The patient sensitive to castor tory personnel served as controls. The test substances were ap- allergens had a significant RAS test ratio of 18.87 to castor bean plied to the skin using both patch and prick tests. Only the castor allergen and a significant RAS test ratio of 8.04 to SLS Castor bean extract and the SLS extract of Castor Wax were evaluated Waxextract. The patient sensitive to peanuts did not have a sig- in the prick test, and the four Castor Wax extracts were evalu- nificant RAS test to either castor bean extract or the Castor Wax ated in the patch test. In the prick test (to detect IgE-mediated extract. The results from this direct RAS study indicate that SLS hypersensitivity), a drop of allergen extract was applied to the Castor Wax extract does contain Castor bean allergens, although skin. Following application to the skin, the underlying superficial the SLS Castor Wax extract can nonspecifically inhibit the RAS layer of the skin was pricked with a 25-gauge needle. Reactions assay (Lehrer et al. 1980). were scored 15 min after application of the allergen. A positive prick test reaction was defined as a wheal and flare reaction with Ricinoleic Acid edema >5 mm. Castor bean extract was tested at concentrations In a retrospective study by Lim and Goh (2000), 202 consec- up to 1000 µg/ml and, SLS Castor Wax extract, at concentra- utive patients (182 females, 20 males) with eczematous cheili- tions up to 22,000 µg/ml. Results from the prick test indicated tis who attended a contact and occupational dermatoses clinic no immediate skin reactivity to castor bean extract or the SLS in Singapore between January of 1996 and December of 1999 Castor Wax extract. were patch-tested. Mean ages for female and male patients were In patch tests (to detect cell-mediated hypersensitivity), a 31.1 and 28.8 years, respectively. Patch tests were conducted semiocclusive patch containing either of the four Castor Wax according to International Contact Dermatitis Research Group extracts (concentration not stated) or paraffin (control) was ap- (ICDRG) recommendations. Twenty-nine patients (2 males,

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CASTOR OIL 67

27 females) had positive reactions to Ricinoleic Acid. Of the oleic acid (2% to 7%), linoleic acid (3% to 5%), palmitic acid 29 reactions, 22 were considered relevant positives. (1% to 2%), stearic acid (1%), dihydrostearic acid (1%), and trace amounts of other fatty acyl groups. Allergenicity/Photoallergenicity Hydrogenated Castor Oil consists primarily of glyceryl- trihydroxystearate. The National Formulary has the following Castor Oil requirements for Hydrogenated Castor Oil: free fatty acids (free In a study by Hashimoto et al. (1990), 103 patients (ages not fatty acids in 20 g require for neutralization not more than 11.0 stated) suspected of having cosmetic-related contact dermati- ml of 0.1 N sodium hydroxide), heavy metals (0.001%), hy- tis were patch tested with castor oil (as is). Both patch tests droxyl value (between 154 and 162), iodine value (not more and photopatch tests were conducted according to ICDRG basic than 5), and saponification value (between 176 and 182). standards. The patch test units consisted of a combined use of Ricinus Communis (Castor) Seed Oil is produced by cold both Finn chambers and Scanpor tape. One patient had a posi- pressing of the seeds of Ricinus communis and subsequent clar- tive patch test reaction to castor oil. No positive photopatch test ification of the oil by heat. Castor Oil does not contain ricin reactions were reported. because ricin does not partition into the oil, due to its water- solubility. Hydrogenated Castor Oil is the end product of con- Case Reports trolled hydrogenation of Ricinus Communis (Castor) Seed Oil. Case reports involving Castor Oil and Ricinoleic Acid esters One of the simplest methods for obtaining Ricinoleic Acid is are summarized in Table 6. the hydrolysis of castor oil. Ricinus Communis (Castor) Seed Oil and Glyceryl Ri- Occupational Exposure cinoleate absorb UV light, with a maximum absorbance at Göhte et al. (1983) conducted a study involving 28 persons 270 nm. (14 men, 14 women; age range = 17 to 65 years) who were em- Reportedly, Ricinus Communis (Castor) Seed Oil and Hy- ployed by a company involved with importing, preparing, and drogenated Castor Oil were used in 769 and 202 cosmetic prod- distributing plant products that are used in spices and as ingre- ucts, respectively, in 2002. The following use frequencies were dients of health foods. The period of employment varied from reported for Ricinoleic Acid and its salts and esters: Glyceryl two months to 20 years, and 25 of the 28 were smokers. Thir- Ricinoleate (16), Ricinoleic Acid (6), Sodium Ricinoleate (12), teen of the 28 developed the following work-related symptoms: Zinc Ricinoleate (3), and Cetyl Ricinoleate (55). rhinitis, conjunctivitis, asthma, itch, or urticaria. Blood samples The highest reported use concentration for Ricinus Commu- from 25 subjects were subjected to IgE analysis using Phadebas nis (Castor) Seed Oil is associated with lipsticks (81%). Other IgE PRIST (paper radioimmunosorbent test). Phadebas RAST maximum ingredient use concentrations are: Hydrogenated Cas- (radioallergosorbent test) was performed on 25 persons against tor Oil (up to 39%), Glyceryl Ricinoleate (up to 12%), Zinc Ri- antigens from an extract of castor oil bean. Fifteen subjects were cinoleate (up to 2%), Cetyl Ricinoleate (up to 10%), and Octyl- prick-tested and 14 were patch-tested with an extract of castor dodecyl Ricinoleate (up to 5%). oil bean. Of the three tests, only the RAST test yielded positive Potassium Ricinoleate, Ethyl Ricinoleate, Glycol Rici- results; 1 of 25 subjects had an allergic reaction to an extract noleate, Isopropyl Ricinoleate, and Methyl Ricinoleate are not of castor oil bean. None of the subjects without work-related currently reported to FDA as in use, nor are there industry survey symptoms had positive reactions. data indicating a current use concentration. Castor oil is classified by FDA as generally recognized as safe and effective for use as a stimulant laxative. An acceptable SUMMARY daily intake (for man) of 0 to 0.7 mg/kg body weight has been Ricinus Communis (Castor) Seed Oil, Ricinoleic Acid, established by FAO/WHO for castor oil. Glyceryl Ricinoleate, Glyceryl Ricinoleate SE, Potassium Ri- Castor oil is a triglyceride that is hydrolyzed in the small in- cinoleate, Sodium Ricinoleate, Zinc Ricinoleate, Cetyl Rici- testine by pancreatic enzymes, leading to the release of glycerol noleate, Ethyl Ricinoleate, Glycol Ricinoleate, Isopropyl Ri- and Ricinoleic Acid. Ricinoleic Acid was rapidly metabolized cinoleate, Methyl Ricinoleate, and Octyldodecyl Ricinoleate in a study of adult rats fed a diet containing 48.4% castor oil for function, in cosmetics, as skin-conditioning agents, emulsion 4to6weeks. These animals did not have significant amounts stabilizers, and surfactants. Ricinus Communis (Castor) Seed of Ricinoleic Acid in phospholipids of the small intestine, liver, Oil also functions as a fragrance and Hydrogenated Castor Oil and skeletal muscle, nor in glycerides of the liver. A close cor- also functions as a viscosity-increasing agent—nonaqueous. In relation between the dose administered in the diet and the per- the case of Zinc Ricinoleate, functions include deodorant agent, centage of Ricinoleic Acid in the feces (greater absorption of the opacifying agent, and anticaking agent. smaller dose) was noted in rats. Castor oil was readily absorbed Ricinoleic Acid accounts for 87% to 90% of the fatty acyl and metabolized in two groups of male rats that were fed 10% groups of Ricinus Communis (Castor) Seed Oil and the follow- castor oil in the diet (cholesterol-enriched and cholesterol-free, ing other fatty acids comprise the remaining fatty acyl groups: respectively) for 20 days. Fecal recovery of Ricinoleic Acid was

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TABLE 6 Case reports with patch test results for Castor Oil and Ricinoleic Acid esters and salts Subjects Findings/patch test results References 22-year-old female Cheilitis worsened after application of various lipsticks and lip Sai 1983 (with cheilitis) creams. Product concentrations of Castor Oil ranged from 10% to 67%. ++ patch test reactions to Castor Oil and each lipstick or lip cream that contained Castor Oil. Also, ++ patch test reactions to Ricinoleic Acid (as is), 30% Ricinoleic Acid in petrolatum, and purified Castor Oil (as is). No reactions to products that did not contain Castor Oil. 23-year-old female ++ reactions to Castor Oil and make-up remover containing Brandle et al. 1983 (history of facial Castor Oil. eczema) with acute dermatitis of the face 29-year-old female Acute edematous dermatitis on the lips and perioral area, which Andersen and Nielsen (history of spread to entire face and neck, after using a lipstick product 1984 deodorant containing Castor Oil. ++ patch test (Finn chambers, dermatitis) International Contact Dermatitis Research Group [ICDRG] recommendations) results for Castor Oil. 24-year-old female Swelling of lips over 10-month period. Negative patch test results Yoshikawa et al. 1986 for Castor Oil and five brands of lipstick containing Castor Oil. 31-year-old female Developed itchy rashes in the axillae, which spread over the Dooms-Goosens et al. (no history of entire body. Positive patch test reaction (++,at38and96 h) to 1987 dermatitis) a deodorant that was used. Repeated open application test results for Castor Oil and a Zinc Ricinoleate mixture (Zinc Ricinoleate (88%), triethanolamine, zinc resinate, isostearic acid, dipropylene glycol, sodium lactate, and abietic acid) were positive. 45-year-old female Presented with an itchy dermatitis on both axillae, and red Dooms-Goosens et al. (history of papules scattered over face and trunk. Patch test results 1987 intolerance to indicated a red erythematous response to a deodorant that was cosmetics and used. Patch test results for Castor Oil were negative at 72 h and adhesives) +/? at 96 h. Repeated open application test results for Castor Oil were negative after 7 days; ++ patch test results for 20% Glyceryl Ricinoleate (in petrolatum) at 72 and 96 h. Also, ++ reaction to Zinc Ricnoleate mixture (defined in preceding case report) at 96 h. This mixture also produced an erythematous test response in 1 of 10 control subjects. 40-year-old chemist Developed eczema on the knee 10 days after joint surgery. Jost et al. 1989 (history of eczema Positive patch test results for surgical adhesive containing due to chemical Castor Oil. Negative patch test results for Castor Oil. exposure on the job) 20-year-old female Allergic cheilitis (itching, swelling, and redness of the lips) after Fisher 1991 using a total of 16 lipstick products. Positive patch test results for Castor Oil and each product. 17-year-old female Positive patch test reactions to Castor Oil and 3 lipsticks. Fisher 1991 (with lip dermatitis)

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TABLE 6 Case reports with patch test results for Castor Oil and Ricinoleic Acid esters and salts (Continued) Subjects Findings/patch test results References 17-year-old female Developed itchy erythematovesicular reaction on right hand after Lodi et al. 1992 (allergic to metals) using a liquid wart remover containing flexible collodion (contains 1.67% Castor Oil). +++patch test reaction to wart remover. ++ patch test reactions to Castor Oil and 20% Castor Oil in petrolatum. Negative patch test reactions to Castor Oil and 20% Castor Oil in petrolatum in 15 control subjects. 51-year-old female Developed two roundish, erythematous vesicular plaques in left Tabar et al. 1993 gluteal and right anterolateral neck regions (application sites) after 20 days of treatment with an anti-wart solution (flexible collodion, common component of wart removers, contains Castor Oil). ++ patch test reactions to Castor Oil 20% Castor Oil in petrolatum, and the wart remover. 65-year-old male Rash on face, neck, and hands. Same rash appeared 3 years Manzur et al. 1995 (history of rash earlier, following wood cutting. Patch test results for Castor Oil following wood (100%) were negative. cutting in orchard) 30-year-old female Hand dermatitis after exposure to cream containing Castor Oil Wakelin et al. 1996 (history of mild and zinc. +++patch test reaction to Castor Oil. Negative atopic eczema) results for 10 control subjects. 29-year-old female Sore, cracked lips, followed by vesicular facial edema, after use of Tanetal. 1997 (nonatopic) the same lipstick product for several years. ++ patch test (Finn chambers, ICDRG criteria) results for Castor Oil. 24-year-old female Pruritic papules, swelling, and pigmentation on and around lips Inoue et al. 1998 (history of hay after applying lipstick. Negative patch test results for Castor Oil fever and dry lips) (as is). 20-year-old army Rashes developed after applying a camouflage stick containing Goon et al. 1999 recruit 47.1% Castor Oil. A mildly pruritic vesicular eruption appeared on the neck, chin, face, and hands; swelling of eyelid also observed. Symptoms lasted for 3 or 4 days. 43-year-old female Recurrent facial dermatitis and cheilitis after use of a facial Le Coz and Ball 2000 (history of sinus moisturizer containing 8% Castor Oil. +/+ patch test (Finn aspergillosis and chambers) reactions to the moisturizer, Castor Oil, 15% Castor facial edema after Oil in petrolatum, and 10% Castor Oil in petrolatum. + to ++ oral dosing with reactions to several lipstick and lip balm products containing tixocortol pivalate) Castor Oil. 51-year-old female Pruritic erythema in both axillae after use of a deodorant Magerl et al. 2001 containing 76% Zinc Ricinoleate. One week later, developed acute contact dermatitis of the lips after using perfumed lipstick containing Glyceryl Ricinoleate. Lipstick was previously tolerated. + patch test reaction to deodorant and lipstick. + patch test reaction to 30% Glyceryl Ricinoleate in petrolatum. Re-patch testing yielded the following results: 76% Zinc Ricinoleate in petrolatum (?+ reaction on day 3) and no reactions to 30%, 15%, or 1% Zinc Ricinoleate in petrolatum 20% Glyceryl Ricinoleate in petrolatum (+ reaction on day 3), 30% Glyceryl Ricinoleate in petrolatum (+ reaction on day 3), and Castor Oil, as is (no reaction). (Continued on next page)

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TABLE 6 Case reports with patch test results for Castor Oil and Ricinoleic Acid esters and salts (Continued) Subjects Findings/patch test results References 58-year-old female After knee surgery, acute contact dermatitis around surgical Reichert-Pénétratet al. (history of allergy incision rapidly became blistered and spread to right lower limb. 2001 to adhesive tapes) Patch test results for Castor Oil (1% and 10% in petrolatum) were negative. 50-year-old female After hip surgery, acute dermatitis around surgical incision spread Reichert-Pénétratet al. (history of allergy to lower right limb. Patch test results for Castor Oil (1% and 2001 to adhesive tapes) 10% in petrolatum) were negative. 30-year-old female Presented with scaly erythema with associated diffuse Leow et al. 2003 (with pigmented hyperpigmentation and itchy swelling for past one year after contact cheilitis) using various lipsticks. ++ patch test reactions to Ricinoleic Acid at day 3.

approximately 0.5% of the total ingested. 3,6-epoxyoctanedioic Unlike prostaglandin E1 (chemically similar to Ricinoleic acid, 3,6-epoxydecanedioic acid, and 3,6-epoxydodecanedioic Acid), Ricinoleic Acid did not induce dose-dependent stimula- acid were excreted by rats given castor oil intragastrically. These tion of adenylate cyclase in villus cell (from hamster small in- acids were not detected in the urine prior to dosing with castor testine) homogenates. Compared to saline-treated controls, cas- oil. tor oil significantly reduced the infiltration of leukocytes and Following the oral administration of Hydrogenated Castor the prostaglandin E2 (inflammatory mediator) content of the rat Oil to rats at dietary concentrations of 0.865% and 8.65%, the aqueous humor and iris-ciliary body. deposition of hydroxystearic acid in the abdominal fat and other Sodium Ricinoleate significantly inhibited (considered mod- body lipids was reported. In all cases, hydroxystearic acid was est; p <.05) cytokine-induced endothelial adhesion molecule accompanied by hydroxypalmitic acid, hydroxymyristic acid, expression when incubated with human saphenous vein endothe- and hydroxylauric acid (hydroxystearic acid metabolites). Rici- lial cells. In a similar experiment, Ricinoleic Acid significantly noleic Acid or Methyl Ricinoleate administered by gastric intu- decreased IL-4-induced VACM-1 expression (50% inhibitory bation to male rats resulted in peak absorption of Ricinoleic Acid concentration between 10 and 100 µM). Neither cell number within 30 min post dosing in the following fractions: triglyceride, nor viability was increased in this concentration range. Rici- diglyceride, monoglyceride, and free fatty acid. noleic Acid did not inhibit TNFα-induced vascular cell adhesion Castor oil (labeled with 131I) was administered to five hy- molecule-1 expression to any significant extent. Castor oil in the pertensive patients on the day before a scheduled intravenous diet induced a significant increase in prostaglandin E2 concen- pyelogram, with results demonstrating that castor oil can be ab- trations in tissues of the intestinal mucosa, placenta, amnion, sorbed by human subjects, that absorption is inversely related to and amniotic cells in groups of pregnant Wistar rats. the administered dose, and, at small doses (4 g), that absorption Castor oil did not have antimicrobial activity in the Staphylo- is virtually complete. coccus aureus, Escherichia coli,orCandida albicans bacterial Following oral administration of castor oil (10 to 15 ml) to strains. There was no correlation between Sodium Ricinoleate three healthy subjects, the following three epoxydicarboxylic antibacterial activity against cells in suspension and antiplaque acids were excreted in the urine: 3,6-epoxyoctanedioic acid; activity. 3,6-epoxydecanedioic acid; and 3,6-epoxydodecanedioic acid. Following the oral administration of castor oil (2 ml), macro- Castor oil caused a marked decrease in the transepidermal scopic damage, characterized mainly by vasocongestion, was water loss rate from full-thickness, cartilage-stripped skin from observed throughout the duodenum and jejunum in rats. After the ventral ear of the male Syrian golden hamster. In another male albino rats were dosed intraperitoneally with 100 mg Ri- study, Ricinoleic Acid enhanced the in vitro transdermal perme- cinoleic Acid, all animals were dead within 24 h. The acute oral ation of 5-fluorouracil in hairless mouse skin. LD50 for Ethyl Ricinoleate exceeded 5.0 g/kg in rats. A single Oral dosing with Sodium Ricinoleate, Ricinoleic Acid, or oral dose (2.5 ml/kg) of castor oil caused generalized hyper- castor oil had both stimulatory and inhibitory effects on smooth emia of the intestinal mucosa up to 72 h post dosing in most of muscle (canine small bowel) contraction in vivo. Sodium Ri- 12 ponies. An acute oral LD50 of >10 g/kg was reported for cinoleate, but not Methyl Ricinoleate, depressed the smooth Hydrogenated Castor Oil in a study involving rats. The acute muscle twitch response in a guinea pig ileum preparation at dermal LD50 for Ethyl Ricinoleate exceeded 5.0 g/kg in rabbits. concentrations ranging from 1.25 × 10−5 to 4 × 10−4 M. One of the rabbits died. The skin reactions observed included

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CASTOR OIL 71 moderate erythema in six rabbits, moderate edema in five rab- female albino rabbits daily for 30 days induced slight erythema bits, and slight edema in one rabbit. and edema. Both open skin irritation and repeated application (8 In an acute intravenous toxicity study in which eight mongrel weeks) tests for undiluted castor oil and 10% aqueous castor oil dogs were dosed with castor oil (single bolus dose = 0.1 ml/kg), were well tolerated by rabbits in another study. Slight reddening none of the animals died. Erythema was the only clinical sign of the application site (flank) was observed in a study in which that was reported. An increase in blood histamine occurred in six albino guinea pigs and six young pigs received applications one animal. of undiluted castor oil on 10 successive days. The short-term subcutaneous administration of a vehicle con- In a predictive prick test in which seven normal subjects sisting of castor oil and benzyl benzoate to groups of five were tested with an SLS extract of Castor Wax (concentra- C57Bl/6J mice caused morphological alterations suggestive of tions up to 22,000 µg/ml), no immediate skin reactivity was stress-induced changes. Using electron microscopy, the changes observed. None of the seven subjects patch tested with the observed in the adrenal cortex included disruption of parenchy- SLS Castor Wax extract or the remaining three extracts of Cas- mal mitochondria in the zona fasciculata, increase in the size tor Wax (concentrations not stated) had a delayed skin reac- of some of the mitochondria, loss of the vesicular orientation. In a provocative prick test in the same study, three pa- tion of the inner mitochondrial membrane, and diminution of tients with occupational hypersensitivity to castor allergens were the mitochondrial matrix. Mitochondrial changes were also ob- tested with the SLS extract of Castor Wax at concentrations up served in the zona reticularis and zona fasiculata. to 22,000 µg/ml. All three patients had a positive reaction to Intragastric administration of Ricinoleic Acid (single 0.1 ml the SLS Castor Wax extract. None of the three patients patch- dose) to CD-1 specific pathogen–free mice resulted in the ero- tested had a delayed reaction to the SLS Castor Wax extract or sion of duodenal villi. In another study, perfusion of the rabbit the remaining three extracts of Castor Wax (concentrations not colon with Sodium Ricinoleate solution (5.0, 7.5, and 10 mM) stated). led to mucosal damage (i.e., the loss of epithelial cells). A dose- The positive skin reactions (immediate skin reactivity) in sen- response relationship for mucosal damage was noted. sitized individuals, together with the positive PCA reactions in In an NTP subchronic oral toxicity study, the administration mice and positive direct RAST for reaginic castor allergens that of diets containing up to 10% castor oil to groups of male and fe- were induced by SLS Castor Wax extract, indicate that allergens male F344/N rats continuously for 13 weeks was not associated are present in Castor Wax at low concentrations. with any toxicities. Neither erythema nor edema was observed following a single The subchronic oral toxicity of Hydrogenated Castor Oil (di- topical application of Ricinoleic Acid (in peanut oil) to the paws etary concentrations up to 17.3%) was evaluated in female rats of 8 to 10 male Swiss mice. The application of Ricinoleic Acid in a preliminary feeding trial. A reduced growth rate in rats fed (in peanut oil) to the entire eyelid surface of each of six male 8.65% and 17.3% Hydrogenated Castor Oil, respectively, was albino Dunkin-Hartley guinea pigs induced eyelid reddening the only abnormality that was observed. Based on organ weight and edema at doses of 10, 30, or 100 mg. Cetyl Ricinoleate determinations and the results of hematological and microscopic was classified as a non-irritant following application to the skin examinations, no adverse effects were observed. of three male New Zealand albino rabbits. Diluted TEGODEO In a 16-week study, groups of 15 male albino rats were fed HY 77 (trade name mixture that generally contains >50% Zinc diets containing 0.865% or 8.65% Hydrogenated Castor Oil. Ex- Ricinoleate) was applied, under occlusive patches and after patch cept for the reduced growth rate in rats fed 8.65% Hydrogenated removal. Well-defined erythema was observed at 48 and 72 h at Castor Oil, no adverse effects were observed. the abraded sites of all six rabbits and at the intact sites of four The instillation of undiluted castor oil (0.5 ml) into the rabbit rabbits. This mixture did not produce skin sensitization in a study eye resulted in an ocular injury score of 1 (maximum score involving 30 white guinea pigs. possible = 20). In contrast, undiluted castor oil induced slight Castor oil was not comedogenic when applied to the ears of congestion of the iris and conjunctiva in the rabbit eye in another rabbits for 2 weeks, but there was a slight increase in keratin study, but the instillation of castor oil (10 drops daily for 3 weeks) content within the follicle, with essentially no change in the into the eyes of ten rabbits in a third study produced no damage follicular epithelium. to the corneal epithelium or endothelium. Sodium Ricinoleate caused dose-related increases in cyto- Castor oil swabbed onto the dorsum of each of 216 albino toxicity in cultures of epithelial cells that were isolated from or Long-Evans rats daily for five days to 2 weeks produced a the small intestines of male Syrian hamsters. In another study, mild effect on the epidermis. Undiluted castor oil was applied Sodium Ricinoleate did not induce significant toxicity in hu- for 24 h to two areas on the dorsal surface of six albino angora man saphenous vein endothelial cell cultures at concentrations rabbits. Severe skin irritation was observed. Undiluted castor oil ≤25 µM. At concentrations of 0.1% and 1.0% Sodium Rici- applied to the dorsal skin of six male Hartley guinea pigs, six noleate, applied to serosal surface or rat jejunum, a significant male Wistar rats, and six miniature swine produced mild skin reduction in the number of ganglion cells in the myenteric or irritation in guinea pigs and rats, but not in miniature swine. submucosal plexus was noted. This was not true for the lowest Repeated applications of undiluted castor oil to the flanks of five test concentration (0.01%). Castor oil did not have a toxic effect

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72 COSMETIC INGREDIENT REVIEW on the tubular epithelium following intraluminal injection into male New Zealand white rabbits. Subcutaneous administration the kidneys of five male white albino rats. In a study evaluat- of the extract produced anti-implantation, anticonceptive, and ing the nephrotoxicity of cyclosporine, castor oil served as the estrogenic activity in rats and mice. vehicle control. Castor oil served as the vehicle control in a study evalu- Neither castor oil nor Sodium Ricinoleate was mutagenic ating the effect of long-term treatment with ICI 182,780 (an to the following Salmonella typhimurium strains either with or antiestrogen) on the rat testis. In the control group, four male without metabolic activation: TA98, TA100, TA102, TA1535, Sprague-Dawley rats were injected subcutaneously with castor and TA1537. Results were also negative for castor oil in Es- oil (0.2 ml) once per week and then killed 100 days after the first cherichia coli strain WP2/pKM102. Groups of male and female injection. Spermatogenesis appeared normal in each of the four mice that received diets containing 0.62%, 1.25%, 2.5%, 5.0%, control rats. and 10.0% castor oil (in DMSO) for 13 weeks had no evidence Castor oil (2 oz) was used to stimulate labor in 196 patients of induction of micronuclei in peripheral erythrocytes. Castor oil (between 37 and 42 weeks of gestation) with premature rupture did not induce sister chromatid exchanges either with or without of membranes. The need for cesarean sections was nearly three metabolic activation. Sodium Ricinoleate interfered with cell di- times greater in the control group (15.7% incidence) than in vision in an Escherichia coli strain, causing the organisms to de- patients dosed with castor oil (5.6% incidence). No relationship velop as filaments. Ricinoleic Acid was inactive as a stimulator between dosing with Castor Oil (administered as a 60-ml dose of DNA synthesis and inducer of ornithine decarboxylase ac- in orange or apple juice), birth weight, and mode of delivery was tivity in the rat colon. Concentrations up to 10 mM resulted in found in a study in which castor oil was used to induce labor in insignificant increases in [3H]dThd incorporation and did not in- 52 pregnant women. The cesarean section rate for women dosed crease the level of ornithine decarboxylase activity above control with castor oil was 19.2% (10 of 52 women), compared to 8.3% values. (4 of 48 controls) in the untreated control group. No adverse None of the 20 mice injected intravaginally with 2% Rici- outcomes for the mother or fetus were identified. noleic Acid (in gum tragacanth) had neoplasms or hyperplastic Mild and transient discomfort and minor epithelial changes lesions of the corpus uteri, cervix uteri, vagina, or perineal skin. were observed in the eyes of nine patients after treatment (in- However, benign lung adenomas were observed in 10 of the stillation) with a castor oil solution four times daily for 15 days. 13 mice dosed with Ricinoleic Acid and in 6 of the 24 vehicle In another study involving 100 patients, the instillation of castor control mice that were killed after the 14th month of dosing. oil affected the cornea and conjunctiva. Specifically, the degen- Neither undiluted castor oil, Ricinoleic Acid, nor Glyceryl eration and death of epithelial cells and continuity breaks in the Ricinoleate was a tumor promoter in a study involving groups epithelium were observed. of ten mice. However, the three test substances induced slight Mild skin irritation was reported in 50 adult males patch tested epidermal hyperplasia in groups of three mice following the with undiluted castor oil (0.05 g applied to back). None of the application of each to a small area of skin in the interscapular 1to20patients, either suffering from or suspected of suffer- region. ing from contact allergy to cosmetic products, patch tested with Castor oil extract had a strong suppressive effect on S180 body 30% Hydrogenated Castor Oil had irritant reactions. Undiluted tumors in male Kunming mice. Its suppressive effect on ARS castor oil rubbed gently onto the right thigh of each of three male ascites cancer was also very strong, with 64% of the ascites subjects on 10 successive days produced, at microscopic exam- tumors in mice being completely cured. Compared to controls, ination of excised skin, clear hyperchromasia and an increase in the life extension rate was more than 136% (p <.0012). the number of cells was observed in the basal cell layer. Slight Groups of mice and rats fed diets containing 0.62%, 1.25%, widening of the granular cell layer was also noted. No skin irri- 2.5%, 5.0%, and 10% Castor Oil continuously for 13 weeks tation was seen with 20% Ethyl Ricinoleate in petrolatum using had a slight decrease in epididymal weight (6% to 7%) in mid- 32 healthy male volunteers in a 48-h closed patch test. and high-dose groups of male rats; however, this finding was No skin sensitization or irritation was seen with 20% Ethyl not dose-related. No effects on any other male reproductive end Ricinoleate in a maximization test using 32 healthy male point (testes weight and epididymal sperm motility, density, or volunteers. In 49 patients with contact dermatitis patch-tested testicular spermatid head count) or female reproductive endpoint (48-h Finn chambers) with castor oil, none had a positive (estrous cycle length, or time spent in each phase of the cycle) reaction. One of the 76 dermatitis patients patch-tested with were noted. Castor Oil had a positive reaction, observed only during the Female Wistar rats injected intramuscularly with castor oil second reading (i.e., the day after the 30-min reading). Of 202 (0.2 ml, single dose) on the first day after estrus had suppressed consecutive patients with eczematous cheilitis who attended ovarian folliculogenesis and anti-implantation and abortive ef- a contact and occupational dermatoses clinic in Singapore, fects. Anticonceptive and estrogenic effects of a methanol 29 patients had positive reactions to Ricinoleic Acid. Of the extract of Ricinus communis var.minor seeds (ether-soluble frac- 29 positive reactions, 22 were considered relevant positives. tion) were studied in adult albino Wistar rats, young albino In an allergenicity-photoallergenicity study, 103 patients sus- Wistar rats, immature Swiss albino female mice, and adult fe- pected of having cosmetic-related dermatitis were patch tested

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CASTOR OIL 73 with castor oil. One patient had a positive patch test reaction. safely in aerosolized products because packaging and use ensure None of the patients had a positive photopatch test reaction. that particulates are not respirable (are greater than 10 µm). The In case reports that included patch test results for castor oil, Panel reasoned that the particle size of anhydrous hair sprays (60 Ricinoleic Acid, or esters of Ricinoleic Acid, both positive and to 80 µm) and pump hair sprays (>80 µm) is large compared negative reactions were reported. Of a group of 25 employees at to the median aerodynamic diameter of 4.25 ± 1.5 µm for a a company involved with the preparation of plant products, 1 em- respirable particulate mass. ployee had an allergic reaction to an extract of the castor oil bean. The CIR Expert panel recognizes that certain ingredients in this group are reportedly used in a given product category, but the concentration of use is not available. For other ingredients in DISCUSSION this group, information regarding use concentration for specific The CIR Expert Panel considered that the available data on product categories is provided, but the number of such products Ricinus Communis (Castor) Seed Oil, Hydrogenated Castor Oil, is not known. In still other cases, an ingredient is not in current Ricinoleic Acid, and salts and esters of Ricinoleic Acid in the use, but may be used in the future. current safety assessment are sufficient for evaluating the safety Although there are gaps in knowledge about product use, the of Glyceryl Ricinoleate and Glyceryl Ricinoleate SE, as well. overall information available on the types of products in which Because Ricinus Communis (Castor) Seed Oil contains Rici- these ingredients are used and at what concentration indicate noleic Acid as the primary fatty acid group, safety test data on a pattern of use. Within this overall pattern of use, the Expert the oil is considered broadly applicable to this entire group of Panel considers all ingredients in this group to be safe. cosmetic ingredients. Overall, the available data demonstrate few toxic effects in acute, subchronic, or chronic toxicity tests. Additionally, there were no genotoxic effects of castor oil in in CONCLUSION vitro or in vivo tests. It was noted that the available data include a The CIR Expert Panel concluded that Ricinus Communis 50-week chronic study in which none of the 20 mice injected (Castor) Seed Oil, Hydrogenated Castor Oil, Glyceryl Rici- with Ricinoleic Acid twice per week had neoplasms or hyper- noleate, Glyceryl Ricinoleate SE, Ricinoleic Acid, Potassium plastic lesions of the corpus uteri, cervix uteri, vagina, or perineal Ricinoleate, Sodium Ricinoleate, Zinc Ricinoleate, Cetyl Rici- skin. UV absorption spectra on Ricinus Communis (Castor) Seed noleate, Ethyl Ricinoleate, Glycol Ricinoleate, Isopropyl Rici- Oil and Glyceryl Ricinoleate indicate maximum absorbance at noleate, Methyl Ricinoleate, and Octyldodecyl Ricinoleate are 270 nm, suggesting that there would be no photosensitization safe as cosmetic ingredients in the practices of use and concen- potential of Glyceryl Ricinoleate or Ricinus Communis (Cas- trations as described in this safety assessment. tor) Seed Oil in human subjects at relevant solar UV exposures. Reactions classified as either significantly irritating or al- REFERENCES lergic were not observed in a maximization test on Ethyl Ri- Achaya, K. T. 1971. Chemical derivatives of castor oil. J. Am. Oil Chem. Soc. cinoleate involving 32 healthy male subjects. Although there 48:758–763. is a paucity of predictive patch test data on Ricinoleic Acid Allegri, R., G. C. Deschel, V. Filippi, M. Gibellini, E. Portioli, and D. Veneri. esters/salts, the Panel agreed that the negative guinea pig sen- 1981. Proposal for the pharmacopoeia: hydrogenated castor oil [translation]. sitization data on a trade name mixture containing more than Boll. Chim. Farm. 120:557–558. Ames, B. N., J. McCann, and E. Yamasaki. 1975. Methods for detecting carcino- 50% Zinc Ricinoleate, in conjunction with the human predic- gens and mutagens with the Salmonella/mammalian-microsome mutagenicity tive patch test data, may be extended to support the safety of test. Mutat. Res. 31:347–364. all of the cosmetic ingredients that are included in this safety Andersen, K. E., and R. Nielsen. 1984. Lipstick dermatitis related to castor oil. assessment. Contact Dermatitis 11:253–254. The Panel did consider the 22 positive reactions to Ricinoleic Aplin P.J., and Eiseo T. 1997. Ingestion of castor oil plant seeds. Med. J. Aust. 167:260–261. Acid in a retrospective study involving 202 consecutive patients Ayorinde, F. O., K. Garvin, and K. 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Non–comedogenic The United States Pharmacopoeia, 27th ed., 348. Rockville, MD: United cosmetics. Cutis 17:344–351. States Pharmacopeial Convention. Gaginella, T. S., and P. Bass. 1978. Laxatives: An update on mechanism of Committee of Revision of the United States Pharmacopeial Convention. 2004b. action. Life Sci. 23:1001–1010. The National Formulary, 22nd ed., 2844. Rockville, MD: United States Phar- Gaginella, T. S., V.S. Chadwick, J. C. Debongnie, J. C. Lewis, and S. F. Phillips. macopeial Convention. 1977a. Perfusion of rabbit colon with ricinoleic acid: Dose-related mucosal Cornell University. 2001. Castor Oil. http://www.ansci.cornell.edu/ injury, fluid secretion, and increased permeability. Gastroenterology 73:95– plants/castorbean.html [accessed October 2003]. 101. Cosmetic, Toiletry, and Fragrance Association (CTFA). 2004. Use concentration Gaginella, T. S., A. C. Haddad, V. L. W. Go, and S. F. Phillips. 1977b. Cyto- data from industry survey. Unpublished data submitted by CTFA, May 2004. toxicity of ricinoleic acid (castor oil) and other intestinal secretagogues on 3 pages.2 isolated intestinal epithelial cells. J Pharmacol Exp Ther 201:259–266. Croci T., M. Landi, X. Emonds-Alt, G. Le Fur, J. P. Maffrand, and L. Man- Gaginella, T. S., S. F. Phillips, R. R. Dozois, and V. L. Go. 1978. Stimulation of ara. 1997. Role of tachykinins in castor oil diarrhea in rats. Br J Pharmacol adenylate cyclase in homogenates of isolated intestinal epithelial cells from 121:375–380. hamsters. Effects of gastrointestinal hormones, prostaglandins, and deoxy- Davis, L. 1984. The use of castor oil to stimulate labor in patients with premature cholic and ricinoleic acids. Gastroenterology 74:11–15. rupture of membranes. J Nurse Midwifery 29:366–370. Gao, J., N.-.H. Sun, F.-Y. Wang, and N. Hao. 1999. Effects of castor oil-diet on De Caterina, R., and W. Bernini. 1998. Structural requirements for inhibition the synthesis of prostaglandin E2 in pregnant rats [translation]. Zhonghua Fu of cytokine-induced endothelial activation by unsaturated fatty acids. J Lipid Chan Ke Za Zhi 34:147–149. Res. 39:1062–1070. Garry, D., R. Figueroa, J. Guillaume, and V. Cucco. 2000. Use of castor oil in De Caterina, R., H. Tanaka, T. Nakagawa, P. J. Hauptman, and P. Libby. 1995. pregnancies at term. Altern Ther Health Med. 6:77–79. The direct effect of injectable cyclosporine and its vehicle, cremophor, on Gennaro, A. R., ed. 1990. Remington’s Pharmaceutical Sciences, 18th ed., 785. endothelial vascular cell adhesion molecule-1 expression. Transplantation Easton, PA: Mack Publishing. 60:270–275. Gluud, B. S., T. Boesen, and M. Norn. 1981. The effect of a fatty vehicle on the cornea and conjunctiva. In vivo staining following instilling of oil [translation]. Ugeskr Laeger 143:2345–2347. 2Available from the Director, Cosmetic Ingredient Review, 1101 Göhte, C. J., G. Wieslander, K. Ancker, and M. Forsbeck. 1983. Buckwheat 17th Street, NW, Suite 412, Washington, DC 20036, USA. allergy: Health food, an inhalation health risk. Allergy 38:155–159.

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Goon, A. T.-J., P. P.-L. Ng, and S.-K. Ng. 1999. Allergic contact dermatitis from Langer, K. H., W. Thoenes, and M. Wiederholt.1968. Light and electron military camouflage. Contact Dermatitis 40:290–291. microscopic investigations of proximal convoluted tubules of the rat kid- Gottschalck, T. E., and G. N. McEwen, Jr., eds. 2004. International Cosmetic ney after intraluminal injection of oil [translation]. Pflugers Arch 302:149– Ingredient Dictionary and Handbook, 10th ed. Washington, DC: CTFA. 165. Grainger, D. N., R. Cross, and J. A. Barrowman. 1982. Effects of various sec- Larsen, S. W., E. Rinvar, O. Svendsen, J. Lykkesfeldt, G. J. Friis, and C. Larsen. retagogues and human carcinoid serum on lymph flow in the cat ileum. Gas- 2001. Determination of the disappearance rate of iodine-125 labelled oils troenterology 83:896–901. from the injection site after intramuscular and subcutaneous administration Grant J, ed. 1972. Hackh’s Chemical Dictionary, 4th ed., 584. New York: to pigs. Int J Pharm 230:67–75. McGraw-Hill Book Co. le Coz, C. J., and C. Ball. 2000. Recurrent allergic contact dermatitis and cheilitis Guillot, J. P., J. Y. Giauffret, and M. C. Martini. 1979. Etude de tolerances due to castor oil. Contact Dermatitis 42:114–115. oculaire et cutanee chez la lapin, de differentes matieres premieres utilisees Lehrer, S. B, R. M. Karr, D. J. G. Müller, and J.E. Salvaggio. 1980. Detection en cosmetologie, et provenant de fabrications diverses (2eme partie). Int. J. of castor allergens in castor wax. Clin. Allergy 10:33–41. Cosmet. Sci. 1:27–57. Leow, Y. H., S. H. Tan, and S. K. Ng. 2003. Pigmented contact cheilitis from Gunstone, F. D. 1989. Chemical reactions of fats and oils. Biochem Soc Trans ricinoleic acid in lipsticks. Contact Dermatitis 49:48–49. 17:1141–1142. Lewis, R. J. Sr. 1997. Hawley’s Condensed Chemical Dictionary, 13th ed., 221, Hachiya, N. 1987. Evaluation of chemical genotoxicity by a series of short-term 749, 922, 972, 1027, 1200. New York: John Wiley & Sons. tests. Akita J Med 14:269–292. Lewis, R. J., Sr., ed. 2000. Sax’s Dangerous Properties of Industrial Materi- Hagenfeldt, L., L. Blomquist, and T. Midtvedt. 1986. Epoxydicarboxylic als,10th ed., 738. New York: John Wiley & Sons. aciduria resulting from the ingestion of castor oil. Clin Chim Acta 161:157– Lide, D. R., and H. P. R. Frederiske, eds. 1993. CRC Handbook of Chemistry 163. and Physics, 74th ed., 3-453. Boca Raton, FL: CRC Press. Hashimoto, Y., T. Sugai, A. Shoji, S. Asoh, K. Watanabe, and A. Inoue. 1990. Lieb, L. M., R. A. Nash, J. R. Matias, and N. Orentreich. 1988. A new in vitro Incidence of positive reactions in patch tests with ingredients of cosmetic method for transepidermal water loss: A possible method for moisturizer products in 1989 and representative cases of cosmetic dermatitis (Translation). evaluation. J Soc Cosmet Chem 39:107–119. Skin Res. 32:115–124. Lim, S. W., and C. L. Goh. 2000. Epidemiology of eczematous cheilitis at Haworth, S., T. Lawlor, K. Mortelmans, W. Speck, and E. Zeiger. 1983. a tertiary dermatological referral centre in Singapore. Contact Dermatitis Salmonella mutagenicity test results for 250 chemicals. Environ Mutagen 43:322–326. 5:3–142. Litvinova, L. B., and T. V. Fedorchenko. 1994. The effect of plant oils on the Hino, N., M. Ikushima-Fujimoto, K. Ueda, A. Kume, and N. Higashi. 2000. female reproductive system [translation]. Eksp Klin Farmakol 57:49–51. Patch test results in 346 patients of suspected cosmetic dermatitis (1993– Lodge, N. J. 1994. Direct vasoconstrictor effects of sandimmune (cyclosporine 1996) [translation]. Environ Dermatol 7:144–153. A) are mediated by its vehicle Cremophor EL: Inhibition by the thromboxane Hui, Y. H., ed. 1996. Bailey’s Industrial Oil & Fat Products. Industrial and A2/prostaglandin endoperoxide receptor antagonist ifetroban. J Pharmacol Consumer Nonedible Products from Oils and Fats, 5th ed., vol. 5., 368. New Exp Ther 271:730–734. York: John Wiley & Sons. Lodi, A., S. Leuchi, L. Mancini, G. Chiarelli, and C. Crosti. 1992. Allergy Ihara-Watanabe, M., T. Shiroyama, G. Konda, H. Umekawa, T. Takahashi, T. to castor oil and colophony in a wart remover. Contact Dermatitis 26:266– Yamada and Y. Furuichi. 1999. Effects of castor oil on lipid metabolism in 267. rats. Biosci Biotechnol Biochem 63:595–597. Lorenz, W., A. Schmal, H. Schult et al. 1982. Histamine release and hypotensive Inoue, A, A. Shoji, and S. Aso. 1998. Allergic lipstick cheilitis due to ester gum reactions in dogs by solubilizing agents and fatty acids: Analysis of various and ricinoleic acid. Contact Dermatitis 39:39. components in cremophor El and development of a compound with reduced International Bio-Research, Inc. 1977a. Delayed contact sensitization (guinea toxicity. Agents Actions 12:64–80. pigs). Compound: “Grillocin HY 77” (undiluted) (Translation). Unpublished Magerl, A., R. Heiss, and P. J. Frosch. 2001. Allergic contact dermatitis from data submitted by CTFA. 18 pages.2 zinc ricinoleate in a deodorant and glyceryl ricinoleate in a lipstick. Contact International Bio-Research, Inc. 1977b. Patch test for primary skin irritation Dermatitis 44:119–121. and corrosivity of the compound: “Grillocin HY 77” (10% soy bean oil). Maier, M., D. Staupendahl, H. R. Duerr, and H. J. Refior. 1999. Castor oil Unpublished data submitted by CTFA. 7 pages.2 decreases pain during extracorporeal shock wave application. Arch Orthop Jensen, P.A., and D. Obrien. 1993. Industrial hygiene. In: Aerosol Measurement. Trauma Surg 119:423–427. Principles, Techniques and Applications, ed. K. Willeke and P.A. Baron, 538– Manzur, F., F. El Sayed, and J. Bazex. 1995. Contact allergy to cinnamic aldehyde 540. New York: John Wiley and Sons. and cinnamic alcohol in Oléophytal. Contact Dermatitis 32:55. Johnsen, M. A. 2004. The influence of particle size. Spray Technology and Masri, M. S., L. A. Goldblatt, F. DeEds, and G. O. Kohler. 1962. Relation of Marketing.November:24–27. cathartic activity to structural modifications of ricinoleic acid of castor oil. J Johnson, C. M., J. M. Cullen, and M. C. Roberts. 1993. Morphologic character- Pharm Sci 51:999–1002. ization of castor oil-induced colitis in ponies. VetPathol 30:248–255. Mathias, J. R., J. L. Martin, T. W. Burns, G. M. Carlson, and R. P. Shields. 1978. Jost, T., Y. Sell, and J. Foussereau. 1989. Contact allergy to Manilla resin. Ricinoleic acid effect on the electrical activity of the small intestine in rabbits. Nomenclature and physico-chemistry of Manilla, kauri, damar and copal J Clin Invest 61:640–644. resins. Contact Dermatitis 21:228–238. McKeon, T. A., G. Q. Chen, and J. T. Lin. 2000. Biochemical aspects of castor Kathren, R. L., H. Price, J. C. Rogers. 1959. Air-borne castor-bean pomace oil biosynthesis. Biochem Soc Trans 28:972–974. allergy; a new solution to an old problem. AMA Arch Ind Health 19:487–489. Meyer, F. U., C. Wollmann, N. Exner, and G. Exner. 1976. Comparative studies Kato, A., and Y. Yamaura.1970. A rapid gas chromatographic method for the on the skin of guinea pigs, swine and man following the effect of various determination of fatty acid compositions of soybean oil and castor oil. Chem external agents [translation]. Dermatol Monatsschr 162:986–991. Ind 39:1260. Migally, N. 1979. Effect of castor oil and benzyl benzoate used as a vehicle for KIC Chemicals, Inc. 2004. Hydrogenated castor oil, standard grade. Specifica- antiandrogens on the adrenal cortex. Arch Androl 2:365–369. tions. http://www.kicchem.com/hcostandard.htm [accessed September 2004]. Mitri, F., G. J. Hofmeyr, and C. J. Van Geldren. 1987. Meconium during labor– Laboratoire De Recherche Et D’Experimentation. 1994. Acute oral toxicity and self-medication and other associations. SAMJ 431–433. skin irritation and/or corrosion tests on cetyl ricinoleate. Unpublished data Mordenti, J. J., R. E. Lindstrom, and J. M. Tanzer. 1982. Activity of sodium submitted by CTFA, April 13, 2004. 2 pages.2 ricinoleate against in vitro plaque. Pharm Sci 71:1419–1421.

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Morehouse, J. L., R. D. Specian, J. J. Stewart, and R. D. Berg. 1986. Translo- Rantuccio, F., D. Sinisi, A. Scardigno, and C. Coviello. 1981. Histological cation of indigenous bacteria from the gastrointestinal tract of mice after oral changes in rabbits after application of medicaments and cosmetic bases. II. ricinoleic acid treatment. Gastroenterology 91:673–682. Contact Dermatitis 7:94–97. Morris, J. A., A. Khettry, and E. W. Seitz. 1979. Antimicrobial activity Rao, M. K., N. Risser, and E. G. Perkins. 1969. The incorporation of ricinoleic of aroma chemicals and essential oils. J. Am. Oil Chem. Soc. 56:595– acid into rat lymph lipids. Proc Soc Exp Biol Med 131:1369–1372. 603. Ratner, B., and H. L. Gruehl. 1929. Respiratory anaphylaxis (asthma) and ricin Morris, W.E., and S. C. Kwan. 1983. Use of the rabbit ear model in evaluating the poisoning induced with castor bean dust. Am J Hyg 10:236. comedogenic potential of cosmetic ingredients. J Soc Cosmet Chem 34:215– Reichert-Pénétrat,S., A. Barbaud, E. Reichert-Pénétrat,F. Granel, and J.-L. 225. Schmutz. 2001. Allergic contact dermatitis from surgical paints. Contact Der- Motoyoshi, K., Y. Toyoshima, M. Sato, and M. Yoshimura. 1979. Compar- matitis 45:116–117. ative studies on the irritancy of oils and synthetic perfumes to the skin Research Institute for Fragrance Materials (RIFM). 2000. Fragrance raw mate- of rabbit, guinea pig, rat, miniature swine and man. Cosmet Toilet 94: rials monographs. Ethyl ricinoleate. FCT 38:S91–S92. 41. Sai, S. 1983. Lipstick dermatitis caused by castor oil. Contact Dermatitis 9:75, Muldoon, D. F., E. A. Hassoun, and S. J. Stohs. 1992. Ricin-induced hepatic 524. lipid peroxidation, glutathione depletion, and DNA single-strand breaks in Salomon, Y., C. Londos, and M. Rodbell. 1974. A highly sensitive adenylate mice. Toxicon 30:977–984. cyclase assay. Anal Biochem 58:541–548. National Academy of Sciences (NAS). 1996. Food Chemicals Codex, 4th ed., Sasol North America, Inc. No date. UV spectrum for castor oil and glyceryl 94 Washington, DC: National Academy Press. ricinoleate. Unpublished data submitted by CTFA, April 14, 2004. 1 page.2 National Toxicology Program (NTP). 1992. NTP technical report on the toxicity Scarpa, A., and A. Guerci.1982. Various uses of the castor oil plant (Ricinus studies of castor oil (CAS No. 8001-79-4) in F344/N rats and B6C3F1 mice communis L.). A review. J Ethnopharmacol 5:117–137. (dosed feed studies). NTIS Report No. PB93151439. Secchi, A. G., M. S. Tognon, and A. Leonardi.1990. Topical use of cyclosporine NTP. 2003. NTP Chemical Repository. Castor Oil. Research Triangle Park, NC: in the treatment of vernal keratoconjunctivitis. Am J Ophthalmol 110:641– NTP. 645. Nikitakis, J. M., and G. N. McEwen Jr., eds. 1990. CTFA Compendium of Shubik, P. 1950. Studies on the promoting phase in the stages of carcinogenesis Cosmetic Ingredient Composition—Descriptions I and II.Washington, DC: in mice, rats, rabbits, and guinea pigs. Cancer Res 10:13–17. CTFA. Simon, B., and H. Kather. 1980. Interaction of laxatives with enzymes of cyclic Ohia, E. O., M. Mancino, and P. S. Kulkarni. 1992. Effects of steroids and AMP metabolism from human colonic mucosa. Eur J Clin Invest 10:231–234. immunosuppressive drugs on endotoxin-uveitis in rabbits. J Ocul Pharmacol Smolinske, S. C. 1992. Handbook of Food, Drug, and Cosmetic Excipients, 70. 8:295–307. Boca Raton, FL: CRC Press. Okui, S., M. Uchiyama, R. Sato, and M. Mizugaki. 1964. Metabolism of hydroxy Song, J. F., C. A. Lau-Cm, and K. H. Kim. 2001. Monohydroxylation and fatty acids. III. Characterization of hydroxy acids in depot fat after feeding of esterification as determinants of the effects of cis- and trans-9-octadecenoic ricinoleic acid. J Biochem 55:81–86. acids on the permeation of hydrocortisone and 5-fluorouraci across hairless Okwuasaba, F. K., U. A. Osunkwo, M. M. Ekwenchi, et al. 1991. Anticonceptive mouse skin in vitro. Int J Pharmaceut 212:153–160. and estrogenic effects of a seed extract of Ricinus communis var. minor. J Srinivasulu, C., and S.N. Mahapatra. 1973. A rapid method for detecting ground- Ethnopharmacol 34:141–145. nut oil in castor oil. J Chromatogr 86:261–262. Oliveira, C. A., K. Carnes, L. R. Franca, and R. A. Hess. 2001. Infertility and tes- Steingrub, J. S., T. Lopez, D. Teres, and R. Steingart. 1988. Amniotic fluid ticular atrophy in the antiestrogen-treated adult male rat. Biol Reprod 65:913– embolism associated with castor oil ingestion. Crit Care Med 16:642–643. 920. Stewart, J. J., and P. Bass. 1976. Effects of ricinoleic acid and oleic acids on the Ordman, D. 1955. An outbreak of bronchial asthma in South Africa, affecting digestive contractile activity of the canine small and large bowel. Gastroen- more than 200 persons, caused by castor bean dust from an oil-processing terology 70:371–376. factory. Int Arch Allergy 7:10. Stewart, J. J, T. S. Gaginella, and P. Bass. 1975. Actions of ricinoleic acid and Paul, H., and C. M. McCay. 1942. The utilization of rats by herbivora. Arch structurally related fatty acids on the gastrointestinal tract. I. Effects on smooth Biochem 1:247–253. muscle contractility in vitro. J Pharmacol Exp Ther 195:347–354. Paulus, G. L., and M. H. Champion. 1972. Gas-liquid chromatographic deter- Stewart, W. C., and R. G. Sinclair. 1945. The absence of ricinoleic acid from mination of castor oil as methyl ricinoleate in lipstick. J Soc Cosmet Chem phospholipids of rats fed castor oil. Arch Biochem 8:7. 23:313–319. Stübiger, G., E. Pittenauer, and G. Allmaier. 2003. Characterization of castor Physical & Theoretical Chemistry Lab. 2004. Safety (MSDS) data for hy- oil by on-line and of-line non-aqueous reverse-phase high-performance liq- drogenated castor oil. http://ptcl.chem.ox.ac.uk/MSDS/HY/hydrogenated uid chromatography-mass spectrometry (APCI and UV/MALDI). Phytochem castor oil.html [accessed September 2004]. Anal 14:337–346. Pinto, A., A. Calignano, N. Mascolo, G. Autore, and F. Capasso. 1989. Castor Tabar, A. I., M. D. Muro, S. Quirce, and J. M. Olaguibel. 1993. Contact dermatitis oil increases intestinal formation of platelet-activating factor and acid phos- due to sensitization to lactic acid and castor oil in a wart remover solution. phatase release in the rat. Br J Pharmacol 96:872–874. Contact Dermatitis 29:49–50. Pöch, G. 1971. Assay of phosphodiesterase with radioactively labeled cyclic Tan, B. B., A. L. Noble, M. E. Roberts, J. T. Lear, and J. S. English. 1997. 35-AMP as substrate. Naunyn-Schmiedeberg’s Arch Pharmacol 268:272– Allergic contact dermatitis from oleyl alcohol in lipstick cross-reacting with 299. ricinoleic acid in castor oil and lanolin. Contact Dermatitis 37:41–42. Polgar, P., and L. Taylor. 1977. Effects of prostaglandin on substrate uptake and Thompson, W. G. 1980. Laxatives: Clinical pharmacology and rational use. cell division in human diploid fibroblasts. Biochem J. 162:1–8. Drugs 19:49–58. Racusen, L. C., B. C. Kone, A. Whelton, and K. Solez.1986. Renal blood TNO BIBRA International Ltd. 1999. Toxicity profile. Castor oil. Surrey, United flow, glomerular filtration rate, and renal morphology in cyclosporine- Kingdom: TNO BIBRA International Ltd. 9 pages. induced acute renal failure in Munich-Wistar rats. Am J Kidney Dis 8:319– Uchiyama, M., R. Sato, and M. Mizugaki. 1963. Characterization of hydrox- 322. yacids in depot fat after feeding of ricinoleic acid. Biochem Biophys Acta Ramsey, J. D., T. D. Lee, M. D. Osselton, and A. C. 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Vieira, C, S. Evangelista, R. Cirillo, R. Terracciano, A. Lippi, C.A. Maggi, and World Health Organization (WHO) .1980. Evaluation of certain food additives. S. Manzini. 2000. Antinociceptive activity of ricinoleic acid, a capsaicin- Twenty-third report of the Joint FAO/WHO Expert Committee on Food Addi- like compound devoid of pungent properties. Eur J Pharmacol 407:109– tives, 1–45. World Health Organization Technical Report Series 648. Geneva: 116. WHO. Vieira, C., S. Tetzer, S. K. Sauer, et al. 2001. Pro- and anti-inﬂammatory ac- Xie, S. A., D. S. Liu, B. L. Li, Z. H. Wang, and J.Y. Tao. 1992. Antitumor effect tions of ricinoleic acid: Similarities and differences with capsaicin. Naunyn- of castor oil extract [translation]. Zhongguo Zhong Yao Za Zhi 17:560–561. Schmiedeberg’s Arch Pharmacol 364:87–95. Yagaloff, K. A., L. Franco, F. Simko, and B. Burghardt. 1995. Essential Wakelin, S. H., A. J. Harris, and S. Shaw. 1996. Contact dermatitis from castor fatty acids are antagonists of the leukotriene B4 receptor. Prostaglandins oil in zinc and castor oil cream. Contact Dermatitis 35:259. Leukotriene Essent. Fatty Acids 52:293–297. Watson, W. C., and R. S. Gordon Jr. 1962. Studies on the digestion, absorption Yahagi, T., M. Degawa, Y.Seino, T. Matsushima, M. Nagao, T. Sugimura, and Y. and metabolism of castor oil. Biochem Pharmacol 11:229–236. Hashimoto. 1975. Mutagenicity of carcinogenic azo dyes and their derivatives. Watson, W. C., R. S. Gordon Jr, A. Karmen, and A. Jover. 1963. The absorption Cancer Lett 1:91–96. and excretion of castor oil in man. J Pharm Pharmacol 15:183–188. Yoshikawa, K., T. Kozuka, T. Doi, Y. Kataoka, K. Fujimoto, and S. Hashimoto. Whiteside-Carlson, V., B. F. Feaster, and W. W. Carlson. 1955. Filament induc- 1986. A case of allergic contact dermatitis to castor oil. 10th Anniversary tion in Escherichia coli by ricinoleic acid. Proc Soc Exp Biol Med 89:382– Meeting of Japan Patch Test Research Group on Contact Dermatitis and Patch 385. Test VII, Kobe, Japan, December 7–8, 1985. Skin Res 28(Suppl. 2):263–265. Wineburg, J. P., and D. Swern. 1973. NMR chemical shift reagents in struc- Zeiger, E., B. Anderson, S. Haworth, T. Lawlor, and K. Mortelmans. 1988. tural determination of lipid derivatives: III. Methyl ricinoleate and methyl Mutagenicity tests: IV. Results from the testing of 300 chemicals. Environ 12-hydroxystearate. JAmOil Chem Soc 50:142–146. Mol Mutagen 11(Suppl. 12):1–158.

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Supplement Panel Book 1 Page 185 • the myristic • • • other opened acid new • safety This In • CIR other chemically The Abstract concluded to exist, safety, use This Keywords ingredients cosmetic used Assessment Ronald Introduction Ronald F. Lillian Salts Final The This 1990, the cetyl aluminum aluminum aluminum aluminum Alan calcium and original Expert safety ester Panel report as safety related myristates the of cosmetics, Expert corresponding this amended skin concentration. acid. myristate, the these C. Panel formulation. A. is C. Andersen, that determined and data similar Report in myristate, Panel addresses assessment safety conditioning Cosmetic safety ingredients safe Becker’, myristates/palmitates, myristate, isostearates/myristates, dimyristate, The this Panel Shank, Hill, additional butyl considered and that safety as myristic are Panel group assessment to assessment the Esters used also myristate PhD2 , have listed butyl the alcohols Ingredient that The was assessment, Expert that PhD 2 , determined agents were combined salts Wilma DVM, of in safety acid, already of structure—activity in myristate, myristic re-reviewed were cosmetics. Cosmetic Curtis Table to and is previously Myristic Panel salts, and are in the of Thomas include a Review as previously many PhD3 ester been the safe this therefore acids, F. sufficient that acid 1. esters reaffirmed along inorganic D. Bergfeld, Ingredient Used The types of expanded cosmetic Amended reviewed. the in other and (CIR) reviewed which myristic Klaassen, 2006 Expert with available j. reviewed includes: of its relationships to esters Slaga, salts Expert cosmetics Acid that salts are support ingredient.’ the to report These Review in acid. Supplement in Panel MD, consider myristic and then that Alkyl salts and data safety by Panel Cosmetics PhD2 , with PhD 2 , esters and the the are Esters re further esters in FACP2 , of in in and (CIR) assessments a determining Safety Panel range of 412, Corresponding 2 Email: Lillian • • • • • • • • • • • • • • • Supplement James Cosmetic Cosmetic Cosmetic are Paul Panel various metabolized. methyl magnesium lauryl isocetyl isobutyl glyceryl isotridecyl isostearyl isopropyl glyceryl glyceryl ethyl isodecyl ethyihexyl decyl Washington, Its Book of cirinfocir-safety.org C. safe Donald concentrations. Becker, W. recognized 1 myristate, Ingredient for Ingredient Ingredient myristate, fatty the myristate, as G. Page myristate, Book myristate, myristate, myristate, isostearate/myristate, dimyristate, myristate, cosmetic Snyder, related myristate, myristate, safety Author: Cosmetic Marks DC myristate, myristate, alcohols myristate, 186 Myristate 1 V. 20036, Review Review Review of Belsito, that ingredients. Ingredient ingredients these Jr, Myristate of USA Scientific Director Expert DVM, salts much myristic MD2 , ingredients Panel Review, readily MD2 , Analyst/Writer of esters Where PhD 2 , acid. in SAGE sagepub.com/journalsPerrnissions.nav Member 29(Supplement © http://ijt.sagepub.com International Reprints DOl: the The the I dissociate TO1177/I09l5818l0374l27 101 are Most Author(s) as data specific current and and 17th used readily Journal permission: of 3) supporting Street, 2010 the 162S-186S in data of in practices hydrolyzed cosmetics. Toxicology esters any NW, did likely Suite not the are of Alkyl Esters Supplement Book 1

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• myristic acid, or stearic acid or cosmetic formulations containing these fatty • myristyl myristate, acids were given to rats orally at doses of 15 to —19g/kg body • octyldodecyl myristate, weight. Most of the data in this assessment was oleic, lauric, • oleyl myristate, palmitic, and stearic acids; myristic acid was included in the • potassium myristate, safety assessment due to its structural similarity. In primary and • propylene glycol myristate, cumulative irritation clinical studies, oleic, myristic, and • sodium myristate, stearic acids at high concentrations were nonirritating. • tetradecyloctadecyl myristate, Cosmetic product formulations containing oleic, lauric, palmi • tridecyl myristate, and tic, and stearic acids at concentrations ranging up to 13% were • zinc myristate. not primary or cumulative irritants, nor sensitizers. On the basis of available data from studies using animals and humans, it is Data from previous safety assessments on butyl myristate, concluded that oleic, lauric, palmitic, myristic, and stearic glycerol myristate, myristic acid, isopropyl myristate and myr acids are safe in current practices of their use and concentration istyl myristate were reviewed and considered during this in cosmetics. assessment. For these ingredients, the previous conclusions CIR Panel (as applicable to the ingredients noted) are summar MyristylMyristateand isopropylMyristate,JACT, 1(4) ized in the following sections. 1982 Acute oral and dermal toxicity tests indicated that myristyl Butyl Myristate,JACT, 9(2) 1990 myristate is nontoxic to rats. This cosmetic ingredient produced Butyl myristate is the ester of butyl alcohol and myristic acid. minimal-to-mild skin irritation, minimal eye irritation in It is a colorless, oily liquid, which is used in cosmetic formula rabbits, and no sensitization in guinea pigs. Studies with rabbits tions at concentrations up to 50%. Aliphatic esters such as butyl indicated that undiluted isopropyl myristate was a mild irritant myristate may be readily hydrolyzed in vivo to the correspond after 24 hours and moderate to severe when applied for3 con ing alcohol and acid, which are then further metabolized. The secutive days. Isopropyl myristate was minimally irritating to median lethal dose 50(LD of butyl myristate was greater than the rabbits’ eyes and was not a skin sensitizer in studies with 8 g/kg in rats. In animal) tests, undiluted butyl myristate was guinea pigs. In limited studies, isopropyl myristate was not moderately irritating but was not a skin sensitizer. No evidence carcinogenic on the skin of mice, but a mixture of isopropyl of eye irritation was noted. On the basis of the available data myristate and isopropyl alcohol significantly accelerated the presented in this report on butyl myristate, as well as other carcinogenic activity of benzo(a)pyrene on the skin. related myristate compounds, the CIR Expert Panel found butyl Human studies with isopropyl myristate indicated that it was myristate safe for cosmetic formulation usage. not a human skin irritant or sensitizer when applied in a product formulation containing 15% to 58% of the ingredient. A prod uct containing 43% of isopropyl myristate produced no photo- GlycerylMyristate, IJT,23(suppl 2:55-94)2004 toxicity and no photocontact allergenicity in human studies. The safety of 43 glyceryl monoesters listed as cosmetic ingre From the available information, it is concluded that myristyl dients was reviewed in a safety assessment completed in 2000. myristate and isopropyl myristate are safe as cosmetic ingredi Glyceryl myristate was included in this group. Glyceryl ents in the current practices of their use. monoesters have little, acute or short-term toxicity in animals, Summaries of the data from these reports are provided in and no toxicity was noted following chronic administration of a italics where applicable throughout the report. mixture consisting mostly of glyceryl di- and monoesters. Undiluted glyceryl monoesters may produce minor skin irrita tion, especially in abraded skin, but in general these ingredients Chemistry are not irritating at concentrations used in cosmetics. These Definition ingredients are not photosensitizers. Glyceryl monoesters and Structure tested failed to produce any significant positive reactions at The definitions, structures, and function in cosmetics of myris concentrations used in cosmetics. Based on these data, the CIR tic acid and the related salt and esters are given in Table 2. Expert Panel found glyceryl myristic safe as a cosmetic ingre Also, included in Table 2 are the formulas/structures and dient in the current practices of its use andconcentration. functions in cosmetics as given in the International Cosmetic Ingredient Dictionary and Handbook.’ The myristates are esters and salts of myristic acid that have the general formula MyristicAcid,JACT, 6(3) 1987 5 shown in Figure i. Oleic, lauric, palmitic, myristic, and stearic acids were reviewed as part of a group. These fatty acids are absorbed, According to the International Cosmetic Ingredient Diction digested, and transported in animals and humans. Little acute ary and Handbook,’ myristic acid (CAS No 544-63-8) is toxicity was observed when oleic, lauric, palmitic, myristic, an organic acid also known as tetradecanoic acid. 5 Supplement Panel Book 1 Page 187 Alkyl Esters Supplement Book 1

164S Internationaljournal of Toxicology29(Supplement 3)

Table I. Related Ingredients Previously Reviewed by the CIR Expert Panel

Ingredient Uses Use Concentrations Conclusion Reference n-Butyl alcohol 112;29 (addendum) 0.I%-I0%; 0.000007%-15% Safe in nail preparations in the current 2,3 practices of use. Cetyl alcohol 2694; 2931 >0.1 %-50%;0000002%- 15% Safe as cosmetic ingredients in the current 4,5 (re-review) practices of use. Glyceryl dimyristate None reported None reported Safe as cosmetic ingredients in the practices of 6 use and concentration as described in this safety assessment. Glyceryl isostearate! None reported None reported Safe as cosmetic ingredients in the current 7 myristate practices of use and concentration.

Glyceryl myristate 19 I%-6% Safe as cosmetic ingredients in the current 7 practices of use and concentration.

Isopropyl myristate 2198; 881 (re-review) 0. I%->50%;0.00008%-78% Safe as cosmetic ingredients in the current 6,8 practices of use. Isostearyl alcohol 41; 16 (re-review) >0.l%-50%; 0.00 l%-50% Safe as cosmetic ingredients in the current 5,9 practices of use. Methyl alcohol 4 0.1%-5% Safe as used to denature alcohol used in 10 cosmetic products.

Myristic acid 36; 73 (re-review) >0. I%-50%;0.00001%-38% Safe in the current practices of use and 2,1 concentration in cosmetics.

Myristyl myristate 160; 244 (re-review) 0. I%-25%;0.01%-20% Safe as cosmetic ingredients in the current 6,8 practices of use.

Oleyl alcohol 1018; 343 (re-review) 0. I%->50%;0.002%-I 8% Safe as currently used in cosmetics. 6,12

Propylene glycol I I None reported Safe as cosmetic ingredients in the current I3 myristate practices of use.

Abbreviation:CIR,Cosmetic IngredientReview.

Aluminum dimyristate (CAS No 56639-51-1) is also known Glyceryl dimyristate (CAS No 53563-63-6) is also known as as aluminum hydroxybis (tetradecanoate) and tetradeca dimyristin; glycerol dimyristate; and tetradecanoic acid, noic acid, aluminum complex. diester with 1,2,3-propanetriol. Aluminum Isostearates/Myristates (no CAS No) is also Glyceryl isostearate/myristate (no CAS No) is also known known as aluminum triisostearate/trimyristate. as glyceryl monoisostearate monomyristate. Aluminum myristate (CAS No 4040-50-0) is also known as Glyceryl myristate (CAS Nos 589-68-4 and 27214-38-6) is aluminum monomyristate; myristic acid, aluminum salt; also known as glycerin monomyristate; glycerol mono and tetradecanoic acid, aluminum salt. myristate; glyceryl monomyristate, monomyristin; myris Aluminum myristates/palmitates (no CAS No) is also tic acid monoglyceride; and tetradecanoic acid, monoester known as aluminum trimyristate/tripalmitate. with 1,2,3-propanetriol. Butyl myristate (CAS No 110-36-1) is also known as butyl Isobutyl myristate (CAS No 25263-97-2) is also known as n-tetradecanoate; myristic acid, butyl ester, and tetrade 2-methyipropyl tetradecanoate; myristic acid, isobutyl canoic acid, butyl ester. ester; and tetradecanoic acid, 2-methylpropyl ester. Calcium myristate (CAS No 15284-51-2) is also known as Isocetyl myristate (CAS No 83708-66-1) is also known as calcium tetradecanoate; myristic acid, calcium salt; and myristic acid, isocetyl ester; tetradecanoic acid, isocetyl tetradecanoic acid, calcium salt. ester; and tetradecanoic acid, isohexadecyl ester. Cetyl myristate (CAS No 2599-01-1) is also known as hex Isodecyl myristate (CAS Nos 17670-91-6 and 51473-24-6) adecyl myristate; hexadecyl tetradecanoate: myristic is also known as 3,7-dimethyloctyl myristate; acid, cetyl ester; myristic acid, hexadecyl ester; and pal isodecyl tetradecanoate; myristic acid, isodecyl mityl myristate, and tetradecanoic acid, hexadecyl ester. ester; tetradecanoic acid, 3,7-dimethyloctyl ester; Decyl myristate (CAS No 41927-71-3) is also known as tetradecanoic acid, isodecyl ester; and tetrahydrogeranyl decyl tetradecanoate; myristic acid, decyl ester; and tet myristate. radecanoic acid, decyl ester. Isopropyl myristate (CAS No 110-27-0) is also known as Ethyl myristate (CAS No 124-06-1) is also known as ethyl 1PM; isoproylis myristas; isopropyl tetradeconoate; tetradecanoate and tetradecanoic acid, ethyl ester. 1-methylethyl tetradecanoate; myristic acid, isopropyl Ethylhexyl myristate (CAS No 29806-75-5) is also known ester; and tetradecanoic acid, 1-methylethyl ester. as 2-ethylhexyl myristate; octyl myristate; and tetradeca Isostearyl myristate (CAS No 72576-81-9) is also known as noic acid, 2-ethyihexyl ester. tetradecanoic acid, isooctadecyl ester. Supplement Panel Book 1 Page 188 Alkyl Esters Supplement Book 1

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Table 2. Definition, Structure, and Function of Myristic Acid and Its Salts and Esters Included in This Report as Given in the International Cosmetic lngreedient Dictionary and 4Handbook’ Ingredient Definition Formula/Structure Function Fragrance ingredient, opacifying Organic acid that conforms agent, Myristic acid generally to the formula: CH(CHCOOH surfactant—cleansing agent Salts ), Aluminum dimyristate Aluminum salt of myristic 32COO]AIOH Anticaking agent; acid [CH(CH emulsion stabilizer; viscosity increasing 32), agent—nonaqueous Aluminum isostearates/ Aluminum salt of a mixture None provided Anticaking agent; emulsion myristates of stabilizer; isostearic acid and myristic viscosity increasing acid agent—nonaqueous Aluminum myristate Aluminum salt of myristic [CH(CHC00] Anticaking agent; emulsion acid 12A1 stabilizer; viscosity increasing 32 agent—nonaqueous Aluminum myristates/ Aluminum salt of a mixture None provided. Anticaking agent; emulsion palmitates ) of stabilizer; palmitic acid and myristic viscosity increasing acid agent—nonaqueous Calcium myristate Calcium salt of myristic CHO1428 . Y2Ca anticaking agent; emulsion acid 2 stabilizer; viscosity increasing agent—nonaqueous Magnesium myristate Magnesium salt of myristic [CH(CH CO0] 2Mg Anticaking agent; slip modifier; acid 32 2 viscosity ) increasing agent—nonaqueous Potassium myristate Potassium salt of myristic C(C12 COOK Surfactant—cleansing agent acid 32 surfactant—emulsifying ) agent Sodium myristate Sodium salt of myristic acid CH(CH COONa Surfactant—cleansing agent; 32 2 surfactant—emulsifying agent Zinc myrist.ate Zinc salt of myristic acid ) Anticaking agent; slip modifier; [CH(CHC0O] 2Zn viscosity increasing ), agent—nonaqueous Esters 32 Butyl myristate Ester of butyl alcohol and II skin-conditioning agent—emollient myristic acid )31C12CH(CH 02H0C Cetyl myristate Ester of cetyl alcohol and Skin-conditioning agentcclusive myristic acid 2),CCH(CH 4H,OC Decyl myristate Ester of decyl alcohol and Skin-conditioning agent—occlusive myristic acid. C12CH](CH CH0CH,(CH )2 0 )283 Ethylhexyl myristate Ester of 2-ethylhexyl alco- CCH(CH,) CHOCHCH(CH) Skin-conditioning hol agent—emollient 32 I and myristic acid CH 23 2CH 3 0

Ethyl myristate Ester of ethyl alcohol and II Fragrance ingredient; hair myristic acid 12CH(CH CHOCH conditioning agent; 32C 23 skin-conditioning ) 0 agent—emollient OH II Glyceryl dimyristate Diester of glycerin and CH(CH OCHCHCH — ).,CHC(CH Skin-conditioning agent—emollient myristic acid C O 32 2 II23 ) 0 Supplement Panel Book 1 Page 189 (rnr1tin,,prl Alkyl Esters Supplement Book 1

I66S International journal of Toxicology 29(Supplement 3)

Table 2 (continued)

Ingredient Definition Formula/Structure Function Fragrance ingredient, opacifying Organic acid that conforms agent, Myristic acid generally to the formula: 12CH(CHCOOH surfactant—cleansing agent Glyceryl isostearate/ Monoester of glyceryn None provided. Skin-conditioning myristate esterfied 32 Agent—emollient; with a blend of isostearic ) surfactant—emulsifying agent and myristic acids O OH Monoester glycerin and Glyceryl myristate of I Skin-conditioning myristic acid 12CH(CH OHOCHCHCH agent—emollient; )32C 2 surfactant—emulsifying agent

11 isobutyl myristate Ester of isobutyl alcohol CH(CH c2 OCH,CHCH Skin-conditioning agent—emollient and )32 3 myristic acid CH3

Isocetyl myristate Ester of isocetyl alcohol Ii Skin-conditioning agent—occlusive and myristic acid C12CH(CH H16330C )32 0 Isodecyl myristate Ester of branched chain Ii Skin-conditioning agent—emollient decyl CH(CH OC and myristic 1021 alcohols acid 32C H Isopropyl myristate Ester of isopropyl alcohol ) Binder; fragrance ingredient; 9

and 1 skin-conditioning agent—emollient myristic acid C12CH(CH 23CHOCH )32 0 Isostearyl mryistate Ester of Isostearyl Alcohol II Binder; Skin-Conditioning and myristic acid )32C12CH(CH H18370C Agent - Emollient Isotridecyl myristate Ester of myristic acid and Hair conditioning agent; isotridecyl alcohol CH(CH 13270C skin-conditioning C12 H agent—occlusive )32 0 Lauryl myristate Ester of lauryl alcohol and Hair conditioning agent; myristic acid CH(CH O(CH skin-conditioning )32C 213CH agent—occlusive 0 Methyl mryistate Ester of methyl alcohol and II) Frangrance ingredient; myristic acid CH(CH OCHJ skin-conditioning 32C agent—emollient ) 0 Myristyl myristate Ester of myristyl alcohol II Skin-conditioning agent—occlusive and CH(CH CHO(CH myristic acid 32C 2313 Octyldodecyl Myristate Ester of octyldodecanol ) 0 ) Skin-conditioning agent—occlusive and myristic acid. II )2C12CH(CH OCHCH(CH,),CH 23 27(CHCHJ ) (continued)

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Table 2 (continued)

Ingredient Definition Formula/Structure Function Fragrance ingredient, opacifying Organic acid that conforms agent, Myristic acid generally to the formula: CH(CHCOOH surfactant—cleansing agent Oleyl myristate Ester of oleyl alcohol and ), 0 Hair conditioning agent; myristic acid 32 II skin-conditioning )2C12CH,(CH )28CH0(CH )27CH,CH(CH agent—occlusive OH Propylene glycol Ester of propylene glycol II Skin-conditioning myristate and CH(CH OCHCHCH agent—emollient; myrisitic acid )32C 23 surfactant—emulsifying agent Tetradecyloctadecyl Ester of tetradecyloctade- ‘j Binder; emulsion stabilizer; film myristate canol II former; and myristic acid 2CH,(CH 1C — OCHCH(CH 5CH, opacifying agent; skin-conditioning agent—occlusive ) 2 I ) CH(CH 13 o )23 Tridecyl myristate Ester of tridecyl alcohol I! skin-conditioning agent—occlusive and myristic acid )32CCH(CH 23),CH(CHOCH

Oleyl myristate (CAS No 22393-93-7) is also known as 9-octadecenyl tetradecanoate and tetradecanoic acid, 9-octadecenyl ester. Potassium myristate (CAS No 13429-27-1) is also known as potassium tetradecanoate and tetradecanoic acid, potas ii sium salt. (CH 0 R Propylene glycol myristate (CAS No 29059-24-3) is also 12CH known as propylene glycol monomyristate; propylene C glycol monotetradecanoate; and tetradecanoic acid, Figure I. General myristate formula,’ in which R may be as small as a 32 monoester with 1,2-propanediol. methyl group for methyl myristate or a potassium ion for potassium Sodium myristate (CAS No 822-12-8) is also known as myristate.) sodium tetradecanoate and tetradecanoic acid, sodium salt. Tetradecyloctadecyl myristate (no CAS No) is also known Isotridecyl myristate (CAS No 96518-24-0) is also known as as myristic acid, tetradecyloctadecyl ester. tetradecanoic acid, isotridecyl ester. Tridecyl myristate (CAS No 36617-27-3) is also known as Lauryl myristate (CAS No 2040-64-4) is also known as tetradecanoic acid, tridecyl ester. dodecyl tetradecanoate; myristic acid, dodecyl ester, and Zinc myristate (CAS No 16260-27-8) is also known as tetra tetradecanoic acid, dodecyl ester. decanoic acid, zinc salt. Magnesium myristate (CAS No 4086-70-8) is also known as tetradecanoic acid, magnesium salt. Methyl myristate (CAS No 124-10-7) is also known as Physicaland ChemicalProperties methyl tetradecanoate; myristic acid, methyl ester; and Myristicacid.Myristic acid occurs as a hard, white, or faintly tetradecanoic acid, methyl ester. yellow, glossy crystalline solid, as a white or yellow-white Myristyl myristate (CAS No 3234-85-3) is also known as 16powder, or as colorless leaflets’ Table 3 presents the phys tetradecanoic acid, tetradecyl ester, and tetradecyl ical and chemical properties7 of of myristic acid and octyldode tetradecanoate. cyl myristate. Octyldodecyl myristate (CAS Nos 22766-83-2 and 83826- Myristic acid is made of tetradecanoic acid (95% 43-1) is also known as myristic acid, 2-octyldodecyl ester; minimum), hexadecanoic acid (4% maximum), and dodecanoic 2-octyldodecyl myristate; tetradecanoic acid, octyldodecyl acid (3% maximum) Cosmetic, Toiletries and Fragrance ester; and tetradecanoic acid, 2-octyldodecyl ester. Association ([CTFA] Table 4)25

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I68S InternationalJournal of Toxicology29(Supplement 3)

Table 3. Physical Properties of Myristic Acid and Octyldodecyl Myristate

Physical Property Value Reference

Myristic acid Molecular weight 228.36 18 228.38 19 Density (g/mL) at 70°C 0.8528 8 Melting point (°C) 58.5 18 58 19 54.4 20 Boilingpoint (°C) 250.5 18 Solubility Water Insoluble 16,18,19 Ethanol Soluble Methanol Very soluble Chloroform Soluble Benzene Very soluble Ether Very soluble Viscosity (cp, at 75 (°C) 5.06 20 Acid value 245.7 20 OctyldodecylMyristate Appearance Oily liquid 21 Test at +8°C Limpid 21 Odor Faint 21 Color (Gardner Scale) 1.5 21 Specific gravity at 20°C 1.435-1.457 21 Viscosity at 20°C 5-45 m.Pa.s 21 Acid value < 7.00 mg KOH/g 21 Saponification value 90-I 10 mg KOH/g 21 Iodine value <7.0 g 12/100g 21 Peroxide value <6.0 meq 02/kg 21 Alkaline impurities <30 ppm NaOH 21 Water content <0.50% 21 Sulphated ashes content <0.1% 21 Heavy metals content 10 ppm 21

Table 4. Comparison of Specifications: Cosmetic and Food Grades of Myristic 22Acid Myristic acid 2223Cosmetics 6Foods’ Iodine value 0.5 maximum 1.0 maximum Acid value 243-249’ 242-249 Saponification value 243-249 242-25 I Unsaponifiable matter 0.2% maximum 1%maximum Arsenic 3 ppm maximum Heavy metals (eg, lead) 10 ppm maximum Residue on ignition 0.1% Titer (solidification point) 52-54°C 48-55°C Water content 0.2% Cosmetic-grademyristicacid specificationsfor fatty acidcompositionis as follows: 2:0,3%maximum;14:0,95%minimum;and 16:0,4%25maximum.

Butyl myristate. Butyl myristate is a light, colorless, oily Isocetylmyristate. Isocetyl myristate is an oily liquid with liquid. It is soluble in acetone, castor oil, chloroform, methanol, practically no odor. It has a density of 0.862, a freezing point mineral oil, and toluene and insoluble in water. Other proper of —39°C,and viscosity of 29.0 at 25°C. It is insoluble in water ties of butyl myristate include a freezing point range of 1°C and soluble in most organic solvents. It is 27combustible. to 7°C, a boiling point range of 167°C to 197°C (at 5 mm Nikko Chemicals Co, Ltd, reported that isocetyl myristate is Hg), and a specific gravity between 0.850 and 0.858 at 2625°C. a colorless liquid with a faint characteristic 28odor. It has a

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Table 5. Frequency of Use and Concentration of Myristic Acid and Its Salts and Esters in Cosmetics

Product Category (Total Number of Products in Each Category (FDA 2008))69 Frequency of 68Use Concentration of Use (%)7071 Myristic Acid Bath products Soaps and detergents (1329) 9 0.1-19 Other (138) 2 Eye makeup

Eye shadow (1196) I 0.5 Mascara (463) 2 Noncoloring hair care products Conditioners (1249) 9 0.00003-0.0002 Shampoos (I 403) 10 0.00002-5 Tonics, dressings, etc (1097) 6 0.00002-I Other (716) 4 Hair-coloring products Color sprays/aerosol (8) 0.00002 Makeup

Blushers (539) — 0.3

Face powders (613) I 0.5 Foundations (635) 15 0.04-0.8

Leg and body paints (29) 2 — Lipsticks (1912) 5 Personal hygiene products

Underarm deodorants (540) I 2 Douches (12) 4 Other (514) 2 69a Shaving products Aftershave lotions (395) 3 0.5 Shaving cream (162) 13 0.5-14 Other (107) 2 Skin care products Skin cleansing creams, lotions, liquids, 101 0.08-15 and pads (1368)

Depilatories (62) — 12

Face and neck creams, lotions, powder I 39497 and sprays (1195) Body and hand creams, lotions, powder 13 0.8-10 and sprays (1513) Moisturizers (2039) 5 0.8 Night creams, lotions, powder and 0.005 sprays (343)

Paste masks/mud packs (418) — 4 Other (1244) 2 8 Suntan products Suntan gels, creams, liquids and sprays (156) — 0.3

Indoor tanning preparations (200) — 2 Total uses/ranges for myristic acid 207 0.00002-20 Aluminum dimyristate Eye makeup Eyeliners (684) Eye shadow (1196) 133 0.2-3 Eye lotions (177) — 0.09 Other (288) b032 Makeup Blushers (539) 6 0.5-2 Face powders (613) 2 0.5-2

Foundations (635) I 0.0 1-2

Makeup bases (164) I

Rouges (99) I3 0.4 Other (406) 4

(continued)

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Table 5 (continued)

Product Category (Total Number of Products in Each Category (FDA 2008))69 Frequency of 68Use Concentration of Use (%)70hI Suntan products Other (62) 2 Total uses/ranges for aluminum dimyristate 174 0.0 1-3 Aluminum myristate Eye makeup Eye shadow (1196) 6 0.01-I Makeup Blushers (539) 14 Face powders (6 13) 3 Total uses/ranges for aluminum myristate 23 0.0 I-I Aluminum myistates/palmitates Makeup Face powders (613) 2 6 Total uses/ranges for aluminum myristates/ 2 6 palmitates Butyl myristate Makeup Lipsticks (1912) 6 Makeup bases (164) 6 Rouges (99) Other (406) 2 Skin care products Moisturizers (2039) Total uses/ranges for butyl myristate 26 Cetyl myristate Eye makeup Eye shadow (1196) Skin care products Face and neck creams, lotions, powder 2 and sprays (1195) Body and hand creams, lotions, powder 6 and sprays (1513) Moisturizers (2039) Other (1244) 2 Total uses/ranges for cetyl myristate 7 6 Glyceryl myristate Fragrance products Other (399) Makeup Makeup bases (164) Personal hygiene products Underarm deodorants (540) Skin care products Face and neck creams, lotions, powder 3 and sprays (1195) Body and hand creams, lotions, powder 5 and sprays (1513) Moisturizers (2039) 5 Night creams, lotions, powder and sprays (343) 2 Paste masks/mud packs (418) 3 Other (1244) 2 Suntan products Suntan gels, creams, liquids and sprays (156) Other (62) Total uses/ranges for glyceryl myristate 25 Isobutyl Myristate Skin care products

(continued)

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Table 5 (continued)

Product Category (Total Number of Products in Each Category (FDA 2008))69 Frequency of 68Use Concentration of Use (%)7071 Body and hand creams, lotions, powder 30 and sprays (1513) Paste masks/mud packs (418) 10 Suntan products Suntan gels, creams, liquids and 3 sprays (156) Total uses/ranges for isobutyl myristate 3-30 lsocetyl myristate Makeup Foundations (635) 5 Skin care products

Other (1244) I Total uses/ranges for isocetyl myristate 6 lsodecyl myristate Makeup

Foundations (635) I

Total uses/ranges for isodecyl myristate I Isopropyl myristate Baby products Lotions, oils, powders, and creams (132) 4 3 Bath products Oils, tablets, and salts (257) 21 39494

Soaps and detergents (1329) I 0.006-I Other (239) 2 23 Eye makeup Eyebrow pencils (147) 12 0.04-20 Eyeliners (684) 49 39495 Eye shadow (1196) 31 39450 Eye lotions (177) 4 — Eye makeup remover (131) 3 — Other (288) 4 Fragrance products Colognes and toilet waters (1288) 9 39461 Perfumes (569) 3 II Powders (278) 3 — Sachets (28) 10 — Other (399) 39 58 Noncoloring hair care products Conditioners (1249) 45 0.5-48

Sprays/aerosol fixatives (371) I 0.02-la Straighteners (144) 4 —

Permanent waves (141) I — Shampoos (1403) 4 0.4-I Tonics, dressings, etc (1097) 39 0.4-23 Other (716) 13 II0c Hair-coloring products 30 Dyes and colors (2481) I d

Shampoos (48) I —

Color sprays (8) I — Bleaches (152) 2 22 Makeup Blushers (539) 36 0.07-2 Face powders (613) 16 0.3-4 Foundations (635) 39 0.001-14

Leg and body paints (29) I — Lipsticks (1912) 49 39472 Makeup bases (164) 8 —

Makeup fixatives (38) I —

(continued)

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Table 5 (continued)

Product Category (Total Number of Products in Each Category (FDA 2008))69 Frequency of 68Use Concentration of Use (%)7071 Other (406) 14 f2_3 Nail care products Basecoats and undercoats (62) Cuticle softeners (18) 2 39537 Creams and lotions (17) 38 Nail polish and enamel removers (41) Other (124) 2 Personal hygiene products

Underarm deodorants (540) 10 0.08-5 I Feminine deodorants (21) 5 39579 Other (514) 13 3-60 Shaving products Aftershave lotions (395) II Preshave lotions (27) 8 17 Shaving cream (162) 5 Other (107) 6 Skin care products Skin cleansing creams, lotions, liquids, 52 39468 and pads (1368) Depilatories (62) 2 Face and neck creams, lotions, powder 48 0.4-5 and sprays (1195) Body and hand creams, lotions, powder 157 2-39 and sprays (1513) Foot powders and sprays (48) 2 Moisturizers (2039) 129 0.2-17 Night creams, lotions, powder and 26 0.1-5 sprays (343) Paste masks/mud packs (418) 10 39521 Skin fresheners (285) 382 3 Other (1244) 61 h Suntan products Suntan gels, creams, liquids and sprays (156) 22 39486 Indoor tanning preparations (200) 6 Other (62) 6 1-3 Total uses/ranges for isopropyl myristate 1057 0.00 1-82 Lauryl myristate Noncoloring hair care products Shampoos (1403) 3 Total uses/ranges for lauryl myristate 3 Magnesium myristate Eye makeup Eyeliners (684) 0.5 Eye shadow (1196) 76 0.6-7 Eye lotions (177) Mascara (463) Other (288) Fragrance products Powders (278) 2 5 Other (399) Makeup Blushers (539) 6 0.2-5 Face powders (613) 80 0.3-10 Foundations (635) 9 0.05-0.09 Lipsticks (1912) 3 Makeup bases (164) 0.0001 Rouges (99) Makeup fixatives (38) 2

(continued)

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Table 5 (continued)

Product Category (Total Number of Products in Each Category (FDA 2008))69 Frequency of 68Use Concentration of Use (%)70.71

Other (406) 9 — Nail care products Nail polishes and enamels (17) Skin care products Body and hand creams, lotions, powder 5 and sprays (1513) Other (1244) Suntan products Indoor tanning preparations (200) Other (62) 2 Total uses/ranges for magnesium myristate 194 0.0001-10 Myristyl myristate Baby products Lotions, oils, powders, and creams (132) 14 39448 Other (138) Bath products Oils, tablets, and salts (257) 5 Eye makeup Eyebrow pencils (147) 6 6 Eyeliners (684) 8 39611 Eye shadow (1196) 8 39575 Eye lotions (177) 5 0.4-4 Other (288) 7 4-6’ Fragrance products Perfumes (569) 39494 Other (399) 6 Noncoloring hair care products Conditioners (1249) 8 Rinses (47) Shampoos (1403) 3 Tonics, dressings, etc (1097) Other (716) — 2’ Makeup Blushers (539) Face powders (613) 0.5 Foundations (635) 7 0.8-5 Leg and body paints (29) 2 39605 Lipsticks (1912) IS 39607 Makeup bases (164) 3 — 37rn Other (406) 5 Nail care products Cuticle softeners (18) 3 Creams and lotions (17) 2 Other (124) 2 Personal hygiene products Underarm deodorants (540) 6 2 Other (514) 3 Shaving products Aftershave lotions (395) 9 2 Shaving cream (162) 7 0.3 Skin care products Skin cleansing creams, lotions, liquids, 4 2 and pads (1368) Face and neck creams, lotions, powder 26 0.5-8 and sprays (1195) Body and hand creams, lotions, powder SI 39449 and sprays (1513) Foot powders and sprays (48)

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Table 5 (continued)

Product Category (Total Number of Products in Each Category (FDA 2008))69 Frequency of 68Use Concentration of Use (%)7071 Moisturizers (2039) 63 0.5-3 Night creams, lotions, powder 10 2 and sprays (343) Paste masks/mud packs (418) 5 0.5 Skin fresheners (285) Other (1244) 6 39449 Suntan products Suntan gels, creams, liquids and sprays (156) 7 Indoor tanning preparations (200) 2 2 Total uses/ranges for myristyl myristate 0.3-17 Octyldodecyl Myristate Baby products Lotions, oils, powders, and creams (132) 2 Eye makeup Eyebrow pencils (147) 0.3 Eyeliners (684) 2 Eye shadow (1196) 3 0.3 Eye lotions (177) 2 Mascara (463) Other (288) Fragrance products Other (399) 2 Noncoloring hair care products Tonics, dressings, etc (1097) Makeup Blushers (539) 2 0.007 Face powders (613) 2 Foundations (635) 8 39518 Lipsticks (1912) 0 0.07-21 Shaving products Aftershave lotions (395) 4 Preshave lotions (27) 2 Skin care products Skin cleansing creams, lotions, liquids, 2 and pads (1368) Face and neck creams, lotions, powder 9 39510 and sprays (1195) Body and hand creams, lotions, powder 8 0.9-4 and sprays (1513) Moisturizers (2039) 16 0.5-2 Paste masks/mud packs (418) 3 Skin fresheners (285) 0.3 Other (1244) 10 Suntan products Indoor tanning preparations (200) 7 Other (62) Total uses/ranges for octyldodecyl myrstate 95 0.007-21 Potassium Myristate Bath products Soaps and detergents (1329) 5 Eye makeup Eyeliners (684) Other (288) Makeup Foundations (635) Skin care products Skin cleansing creams, lotions, liquids, 8 39574 and pads (1368)

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Table 5 (continued)

Product Category (Total Number of Products in Each Category (FDA 2008))69 Frequency of 68Use Concentration of Use (%)7071 Other (1244) Total uses/ranges for potassium myristate 27 5-7 Propylene glycol myristate Eye makeup Other (288) Makeup Lipsticks (1912) 2 5 Other (406) Skin care products Face and neck creams, lotions, powder 4 and sprays (I 195) Body and hand creams, lotions, powder 3 and sprays (1513) Moisturizers (2039) Other (1244) 4 Suntan products Suntan gels, creams, liquids 2 6 and sprays (156) Total uses/ranges for propylene 4-6 glycol myristate Sodium myristate Bath products Soaps and detergents (1329) 3 0.5-6 Noncoloring hair care products Conditioners (1249) Shampoos (1403) 3 Personal hygiene products Underarm deodorants (540) 0.2 Skin care products Skin cleansing creams, lotions, liquids, 6 and pads (1368) Face and neck creams, lotions, powder and sprays (1195) Total uses/ranges for sodium myristate IS 0.2-6 Zinc myristate Eye makeup Eyebrow pencils (147) 4 Eyeliners (684) 5 Eye shadow (1196) 50 0.5-6 Eye lotions (177) 0.05 Other (288) 9 Fragrance products Powders (278) 5 Makeup Blushers (539) 33 0.3-3 Face powders (613) IS 39526 Foundations (635) 5 0.001-6 Lipsticks (1912) 5 Makeup bases (164) 5 Other (406) Nail care products Basecoats and undercoats (62) 0.00005 Nail polishes and enamels (419) 5 Skin care products Face and neck creams, lotions, powder 5 and sprays (1195) Suntan products

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Table 5 (continued)

Product Category (Total Number of Products in Each Category (FDA 2008))’ Frequency of 68Use Concentration of Use (%)70.71

Suntan gels, creams, liquids — 0.1 and sprays (156) Total uses/ranges for zinc myristate 122 0.00005-20 a 6% in a shower gel; 9% in a body scrub. b 0.3% in a lash powder; 2% in a brow powder wax. 1%in an aerosol hair shine; 10%in a hair oil treatment. d 5% after dilution. 11%after dilution. 4% in a lip liner pencil. g 4% in a body scrub. 4% in a foot lotion; 82% in a massage oil. 1%and 3% in tanning oils. 6% in a lash powder. k 4% in an eye pencil. 2% in a hairdressing créme conditioner. “ 7% in a concealer. C 0.7% in a moisturizing sprays. maximum acid value of 1 and a saponification value range of Methods of Manufacture 120 to 130. Aluminum dimyristate, aluminum myristate, butyl myristate, Isopropylmyristate. Isopropyl myristate is a colorless, almost calcium myristate, decyl myristate, ethylhexyl myristate, ethyl odorless, mobile liquid with a bland taste. It is soluble in acet myristate, glyceryl dimyristate, glyceryl myristate, isobutyl myr one, castor oil, chloroform, cottonseed oil, ethanol, ethyl acet istate, isocetyl myristate, isodecyl myristate, isopropyl myr ate, mineral oil, and toluene and insoluble in water, glycerol, istate, isotridecyl myristate, lauryl myristate, magnesium sorbitan, and propylene glycol. It is miscible with liquid hydro myristate,methyl myristate,myristyl myristate,octyldodecylmyr carbons and fixed oils, and it dissolves lanolin, cholesterol, and istate, potassium myristate, propylene glycol myristate, sodium many 293waxes. myristate, tetradecyloctadecyl myristate, tridecyl myristate, and zincmyristatehave plantand syntheticsources.Aluminumisostea Octyldodecyl’ myristate. 2-Octyldodecyl myristate is a color rates/myristates,aluminum myristates/palmitates, cetyl myristate, less, odorless liquid with a maximum acid value of 0.5, saponi glyceryl isostearate/myristate, isostearyl myristate, and oleyl fication value range from 105to 111, and a maximum hydroxyl myristate have plant, animal, and synthetic 15sources. value of 5.0. On ignition, the residue has a maximum of 0.5%.32 22Gattefossé stated that octyldodecyl myristate was Myristicacid.According to the CTFA (now the Personal Care slightly soluble in ethanol at 96°C, soluble in chloroform and Products Council [the Council]), myristic acid is produced methylene chloride, insoluble in water, and freely soluble in commercially by the saponification and fractionation of animal mineral oils. or vegetable fats and oils. The isolated acid fraction is hydro genated to produce the saturated fatty 35acid. Potassium myristate. Potassium myristate is a white-to-pale Myristic acid is a solid organic acid usually obtained from yellow solid with a faint characteristic 33odor. coconut oil, nutmeg butter (Myristica fragrans Houtt), palm seed oils, and milk 18fats. Seed oils of the plant family, Myr istaceae, contain the2 largest amounts ’° of myristic acid (up to UltravioletAbsorption 80%), but small amounts have been measured in most animal myrist Glycer,’I ate. Glyceryl myristate has UV absorption max fats and vegetable oils. of 238 rim and 270 min of 34nm. The following methods have been used in the preparation of myristic acid: isolation from tail-oil fatty acids, from 9- ketotetradecanoic acid; by electrolysis of a mixture of methyl Reactivity hydrogen adipate and decanoic acid, by Maurer oxidation of The myristate esters can be expected to undergo chemical or myristanol; and from cetanol) The most common means of preparation is by fractional8 distillation of hydrolyzed coconut enzymatic hydrolysis to myristic acid and the corresponding alcohol, Transesterification and other typical ester reactions oil, palm kernel 36oil, or coconut 17acids. may also occur. Butyl myristate, if synthesized from a pure, saturated fatty acid, would not significantly autoxidize, disco Butylmyristate. Butyl myristate is derived from the esterifica lor, or develop an odor. tion of myristic acid and butyl alcohol in 35 the presence of an Supplement Panel Book 1 Page 200 Alkyl Esters Supplement Book 1

Becker et ci 177S acid catalyst. The product is stripped to remove excess alcohol Thin-layer chromatography and high-performance liquid and alkali refined to neutralize the catalyst. Butyl myristate is chromatography (HPLC) are also used in fatty acid identifica obtained through fractional 35distillation. tion and 5457quantitation. Methods of detection include UV, fluorescence spectroscopic, and refractive index detection. Isocetylmyristate. Nikko Chemicals Co, Ltd, reported that Mass spectrometry with temperature profiling of the chem isocetyl myristate is produced by the esterification of isocetyl ical ionization source has been reported as a method for initial alcohol and myristic 37acid. compound separation. Its coupling with a second MS allows direct analysis of complex lipid 58sources. Other separation lsopropylmyristate.Isopropyl myristate is commercially pro methods include centrifugal liquid and adsorption chromato duced by distillation, which is preceded by the esterification of 59graphy. Identification procedures range from methods such myristic acid and isopropanol, in the presence of an acid cata as gravimetry and histochemical 60staining to ultraviolet, lyst. The product is stripped to remove excess isopropanol, infrared,4 and nuclear magnetic resonance spectroscopy. ,62 alkali refined to neutralize the catalyst, and then the product Cotte’ et al63 used Fourier-transform infrared2061 (FT-IR) micro is distilled to obtain isopropyl 38myristate. scopy to locate myristic acid in dermal layers.’ Methylmyristate.Methyl myristate is derived by the esterifi cation of myristic acid with methanol or alcoholysis of coconut Impurities oil with 27methanol. It is purified by vacuum fractional Myristicacid.The myristates used as cosmetic ingredients are distillation. mixtures of fatty esters, as the myristic acid and alcohols used in the preparation of these ingredients are themselves mixtures Myristyl myristate. Myristyl myristate is produced by the of acids and alcohols, respectively. The CTFA Cosmetic Ingre esterification of myristic acid and myristyl alcohol in the pres dient Chemical Description for myristic acid lists the follow ence of an acid catalyst. The product is stripped to remove ing as component5 acids: excess myristyl alcohol; alkali is used to neutralize the catalyst, ’ and then purified to separate myristyl 39myristate. • n-tetradecanoic acid, CH(CHC00H (95% minimum), • n-hexadecanoic acid, 12CH(CHC00H (4% maximum), Octyldodecylmyristate. Octyldodecyl myristate is produced • and n-dodecanoic acid,3214CH(CHCOOH (3% maximum). by the esterification of myristic acid with 2-octyl dodecanol, 32 manufactured vegetable Myristic acid may contain) unsaponifiable from 2240sources. 320), material, mostly hydrocarbons, at a maximum) concentration 0.2%, and 4 of some Potassium myristate. Potassium myristate is produced by the grades may contain glyceryl monomyristate at a maximum ’ concentration of 0.07%. Butylated hydroxytoluene (BHT) reaction of potassium hydroxide and myristic 42acid. may Reacting lauric acid, myristic acid, and palmitic acid with be present as an added antioxidant. water, glycerin, potassium hydroxide, and tetrasodium EDTA 5 produces a product containing potassium myristate (15%) as Butylmyristate. Minor’ impurities, which may be present, are well as potassium cocoate (23%), EDTA-4Na (0.2%), and fatty acids (such as myristic acid) at a maximum of water (61.8%). Glyceiylmyristate. Impurities in glyceryl myristate include glycerol (0.3%), diglycerol (0.57%), and free fatty acid (0.14%).6465 The ratio of 1,2-(mono)glycerol diester to total AnalyticalMethods (mono)glycerol diester is 27.8. Specifications include monoe The myristates can be analyzed by thin-layer chromatography ster content (minimum 90%), free glycerol (maximum 1%), 44(TLC), gas-liquid chromatography, and x-ray powder and free fatty acids (maximum 1.5%). The typical value for 46diffraction. 45 heavy metals (as lead) in glyceryl myristate is <10 mg/kg. Two basic methods for the analysis of the fatty acids have been reported by the cosmetic industry. Primarily, gas chroma lsocetylmyristate. Isocetyl myristate is 95% pure with a max tography (GC) of fatty acid methyl esters, prepared by the imum of heavy metals of 20 ppm and arsenic of 2 28ppm. boron trifluoride—methanol method, is used for the separation and relative identification of fatty acids in a 4748mixture. Infra isopropylmyristate. Isopropyl myristate may have myristic red spectra of the fatty acids are used for fingerprinting, func acid, other free fatty acids are present at a maximum concentra tional group identification, and impurity’ 4953screening. tion of 1.0%, and unsaponifiable material is present at a max Determination of physicochemical properties 23also aids in imum concentration of 0.2%. There are pos no known diluents. itive identification of a specific fatty 2047acid. solvents, or additives 38present. Flame ionization detection (FID) is’usually ’coupled with the The ester composition is varied according to the specific GC of fatty acid methyl esters. Mass spectrometry (MS) has usage requirement, provided that the specification limits con also been used with GC for compound identification. form to the following: isopropyl myristate, not less than 54 Supplement Panel Book 1 Page 201 Alkyl Esters Supplement Book 1

178S InternationalJournal of Toxicology29(Supplement 3)

90.0% (limits, ± 5.0%); isopropyl palmitate, not more than Glycerylmyristate. Glyceryl myristate was used in 25 cosmetic 10.0% (limits, ±3.0%); and isopropyl laurate, tridecanoate, products in 2007; no use concentrations were reported, although pentadecanoate, heptadecanoate, and stearate, none more than in 1998, its concentrations ranged from 1% to 6%. 10.0% (limits, 2.0% 66each). CosmeticAerosols Methyl myristate. Technical grade methyl myristate is 93% Cetyl myristate is used in 2 face and neck creams, pure and can be purified to >99.8%.27 Spectrum Chemicals and lotions, powders, and sprays. Laboratory Products stated that a sample of methyl myristate 67 The potential adverse inhaled was 99.4% pure. Impurities were not listed. effects of aerosols depend on the specific chemical species, the concentration, the duration of the exposure, and the site of deposition within the respiratory Myristylmyristate. Myristyl myristate may have free fatty 72system. In general the smaller the particle, the farther into the acids, mainly myristic acid, at a maximum concentration of respiratory tree the particle will deposit and the greater the 1.5%. There are no known diluents, solvents, or additives impact on the respiratory 73system. 39present. Anhydrous hair spray particle diameters of 60 to 80 p.m have been reported, and pump hair sprays have particle diameters of 80 p.m. The mean particle diameter is around 38 p.m in a Octyldodecylmyristate. Nikko Chemicals Co, Ltd stated that 74 32 typical aerosol spray. In practice, aerosols should have at 2-octyldodecyl myristate has a maximum of 20 ppm heavy 75 least 99% of particle diameters in the 10 to 110 p.mrange. This metals and 2 ppm arsenic. means that most aerosol particles are deposited in the nasophar yngeal region and are not respirable. Potassium myristate. Nikko Chemicals Co, 34Ltd stated that potassium myristate has a maximum of 40 ppm heavy metals Noncosmetic and 2 ppm arsenic. Myristic acid is used in foods as a plasticizing, lubricating, binding, and defoaming agent and as a reagent in the manufac Use ture of other food-grade additives.’ Myristic acid is used as a flavoring 3676 agent in foods.’ Cosmetic Straight-chain 7 monobasic6 carboxylic acids from fats and Use information is supplied to the Food oils derived from edible sources,’ such as the fatty acid myristic US and Drug Administra tion (FDA) by industry as part of the Voluntary Cosmetic Ingre acid, are accepted as safe for use in food and in the manufacture dient Reporting Program 68(VCRP). Use concentration of food-grade additives, provided they meet particular condi information is gathered by the Personal Care Products Council tions and specifications. The unsaponifiable matter in the fatty acid or fatty acid-derived food additive must (Council) unless noted otherwise. Table 5 presents the use and not exceed 2%, concentration ofmyristic acid and its salts and esters in cosmetics. the food additive must be free of chick-edema factor, and it There were no uses or use concentrations reported for the must be produced and labeled in accordance with good manu following: facturing 77practice. Butyl myristate is also used as a plasticizer, as a lubricant for • aluminum isostearates/myristates, textiles, and in paper 78stencils. • calcium myristate, Both ethyl and methyl myristate are generally recognized as • decyl myristate, safe food additives by the 79FDA. • ethyl myristate, • ethylhexyl myristate, • glyceryl dimyristate, General Biology • glyceryl isostearate/myristate, • glyceryl myristate, Metabolismand Absorption Myristicacid. other higher • isostearyl myristate, Like molecular weight aliphatic esters, • isotridecyl myristate, the myristates are readily hydrolyzed to the correspond ing alcohols and • oleyl myristate, acids, which are then further 76metabolized. Myristic acid is a digestible constituent • tetradecyloctadecyl myristate, or of most vegetable and animal fats and is • tridecyl myristate. nontoxic when ingested. Rioux et ’8a1 incubated cultured°8 Sprague-Dawley rat hepa tocytes in radiolabeled myristic acid for 3, 6, 12, and 2 hours. Butyl myristate. Butyl myristate was used in 26 cosmetic Electrophoresis of the products revealed that myristic acid products in 2007. Concentration of use data were not reported, (4 nmol/L) was metabolized into 18 well-resolved proteins in although in 1990, concentrations ranged from 1% to 50%.’ the 10 to 20 kd range.

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Cotte et a163 used FT-JR to measure the penetration of Testing of isopropyl myristate showed mixed results regarding pre-deuterated myristic acid in pig ear skin using Franz diffu dermal penetration enhancement. 90-96 sion cells. After 1 day, myristic acid penetrated to the epider mis. For comparison, palmitic acid was detected in the stratum corneum and did not penetrate any further. Other Effects Dermal Ethylmyristate. Savary and Constantin orally administered 82 Isopropyl myristate. Suzuki et reported that ethyl myristate mixed with olive oil in the feed (90% boiled a185 isopropyl rice, 10% lipid by wet weight) of rats with thoracic-duct fistula. myristate induced acanthosis, edematous degeneration of col Lymph was collected for 24 to 100 hours. The ester was recov lagen fibers, and changes in blood vessels when applied to ered in small quantities in the thoracic-duct lymph. In the Angora rabbits. hydrolysis of lymph triglycerides, fatty acid yields from total dietary lipids were 55 mg/h coming from total dietary lipids Enzyme and 22 mg/h coming from dietary monoalcohol fatty ester. Methylmyristate.Osama et a197 reported that the half maxi mal inhibitory concentration 50(IC of methyl myristate for the inhibition of rat brain prostaglandin) D synthase and swine brain Ethyl and methyl myristate. Hydrolysis of ethyl myristate (emulsified in buffer) by rat pancreatic juice or pure porcine prostaglandin 2D dehydrogenase was >200 jimol/L in both pancreatic lipase was at a lower relative rate (25% and 31%, cases. respectively) than tetradecyl butyrate (100% and 110%), hexa decyl fonnate (55% and 80%), hexadecyl propionate (37% and Cytotoxicity 46%), hexadecyl butyrate (100% and 100%), and n-hexyl lau Methylmyristate.Takeara et a198 used the 3-(4,5-dimethylthia- rate (110% and 150%). The relative rates of hydrolysis for zole-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test to methyl myristate were 61% and 90%, respectively. evaluate the cytotoxicity of methyl myristate on 4 strains of leu 83 kemia cells. For acute promyeloblasic leukemia (HL-60) cells, the Isopropylmyristate.Four monkeys were exposed for 5 seconds IC50 was 4.68 (1.52-14.44 confidence interval [CI]) ig/ to the spray of an aerosol antiperspirant containing 14C-labeled mL, >6.25 ig/mL for chronic myelogenic leukemia (K-526) isopropyl myristate. Two animals were killed immediately cells, >6.25 .ig/mL for lymphoblastic leukemia (CEM) cells, and 84 4.31 after exposure, and the other 2 were killed 24 hours later. The (3.66-5.09 DI) ig/mL for T-cell leukemia (Molt-4) cells. distribution of in the exhaled air and in several tissues indicated that only 0.25% of the dose sprayed at the animals was absorbed; about 10% of this reached the lower respiratory Animal Toxicology tract. Some 85% of the absorbed isopropyl myristate was Acute Oral Toxicity eliminated in 24 hours, mainly as exhaled carbon dioxide; very little labeled material reached any tissues other than the lungs. Data from a previous assessment of myristic acid showed that Suzuki et al85 reported that C-labeled isopropyl myristate little acute toxicity was observed at oral doses of 15 to 19 g/kg penetrated into sebaceous glands,4 stratum spinosum, hair infun body weight of 2.2% to 13% myristic acid in 2rats. In an acute dibula, and follicles. ‘ oral toxicity study of butyl myristate in rats, the 50LD was >8g/ Brinkmann and Muller-Goymann used differential scan kg. The acute oral 50LD for undiluted isopropyl myristate is ning calorimetry, wide-angle86 x-ray diffraction, and small- >16 mL/kg in rats and 49.7 mL/kg in mice.’ angle x-ray diffraction to examine human abdominal and breast skin soaked in isopropyl myristate. The authors reported a Butylmyristate. An acute oral toxicity study of butyl myris slight increase in the short distance of orthorhombically tate was conducted using 10 rats (strainlsex not provided). arranged lipids, while that of hexagonally packed lipids Daily observations were made over a period of 14 days. The decreased. The long distance of the lamellar structure was unaf 50LD was >8 g/kg. No data on weights of animals tested, ranges fected. Isopropyl myristate insertion caused a more densely of chemical concentration tested, or responses of individual packed lipid order. The authors suggest that isopropyl myristate rats were 35available. does a lateral insertion into lipophilic areas of the stratum cor Laboratoire de Recherche et d’Experimentation orally neum microstructure with an anchoring of the isopropyl group administered butyl myristate (200099 mg/kg) to male NMRI in the polar region of the layer. EOPS mice (n 5). The mice were observed for 6 days. There was no mortality, and no clinical or behavioral signs were observed. Weight gain was satisfactory. Dermal PenetrationEnhancement Myristic acid has been tested for its ability to enhance the der Ethyl myristate. Food and Drug Research Laboratories, mal penetration of a number of chemicals. In most cases, skin 00Inc,’ orally treated 10 rats (strainlsex not provided) with treated with myristic acid increased dermal 879penetration. 5 g/kg ethyl myristate. Over a 14-day observation period, none Enhanced penetration was also observed by butyl myristate. of these animals died.

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180S InternationalJournal of Toxicology29(Supplement 3)

Acute Dermal Toxicity only minimal eye irritation in rabbits. Myristic acid (1.5%) was minimally irritiating to the eyes of rabbits. Butyl myristate (2g/kg) was nontoxic and nonirritating when 2 applied to the skin of rabbits.’° 1 Dermal Sensitization Ethyl myristcite. Food and Drug Research Laboratories, Previous safety assessments cited data showing that butyl myr 00Inc,’ dermally treated 10 rabbits with 5 g/kg ethyl myristate. istate was a moderate skin irritant when intradermaly adminis Over a 7-day observation period, 2 10 animals of died. tered to guinea pigs but was not a sensitizer. Isopropyl myristate did not produce sensitization in1guinea pigs. Myristyl Isopropylmyristate. The acute dermal toxicity of undiluted myristate produced minimal skin irritation but no sensitization isopropyl 3 formulations myristate and product containing iso in guinea pigs administered myristyl myristate topically or propyl myristate were evaluated. Isopropyl myristate was intracutaneously. considered nontoxic to the animals tested (rabbits and guinea pigs). Comedogenicity Isopropyl and myristylmyristate. Treatment with isopropyl AcuteParenteral Toxicity myristate resulted in comedogenic activity in the rabbit ear 102-104 Previous safety assessments noted that the intraperitoneal and assay. subcutaneous 20LD for isopropyl myristate exceeded 79.5 Nguyen et 5al’° applied myristyl myristate (50% in petrola mL/kg in rats and the intraperitoneal 50LD exceeded 50.2 turn; 0.5 g) and isopropyl myristate (50% in various mediums; mL/kg in mice. 0.5 g) to the glabrous inner portion of both ears of New Zealand 1 white rabbits (n 6; male 6 = and female; weeks old) for 5 days per week (Monday to Friday) for 4 consecutive weeks. The ears Sub-ChronicDermal Toxicity were then biopsied and scored for comedones through clinical Previous safety assessments noted that myristic acid produced examination and slide biopsy. The control substance was crude slight irritation after topical application to the skin of the exter coal tar (10%). Isopropyl myristate was found to be comedo nal ear canal of 4 albino rabbits. No adverse effects were pro genic in all media; myristyl myristate was less comedogenic. duced from subchronic topical application of myristic acid to rabbit 2skin. Subchronic dermal toxicity studies with product formula Genotoxicity tions containing 16% to 47% isopropyl myristate showed no Isopropylmyristate. Blevins and 06Taylor’ reported that iso toxicity over 4 weeks.’ Butyl myristate and isopropyl myristate propyl myristate tested negative in the Salmonella/microsome were nontoxic when applied to the skin of rabbits. Isopropyl test in strains TA1538, TA1537, TA1535, TA100, and TA98, myristate was moderately-to-severely irritiating when applied with and without activation. for 3 consecutive days to the clipped skin of rabbits. Butyl myr istate was considered moderately irritating in rabbits in one study and nonirritating in another. Carcinogenicity Previous safety assessments noted that isopropyl myristate was InhalationToxicity not carcinogenic on the skin of mice, but a mixture of isopropyl myristate and isopropyl alcohol significantly accelerated the Previous safety assessments cited acute inhalation toxicity carcinogenic activity of benzo(a)pyrene on the skin.’ studies in rats showing no adverse effects from 2 product for mulations containing 16% to 20% isopropyl myristate.’ No toxic effects were observed in subchronic inhalation toxicity Clinical Assessment of Safety studies in guinea pigs and in cynomolgus monkeys. Previoussafety assessments on the followingingredients are summarized below: ChronicToxicity Isopropylmyristate. ‘ Human primary skin irritation studies No chronic toxicity data were found. showed no reactions to isopropyl myristate alone and a mild irritation from product formulations containing 15% to 58% isopropyl myristate. Repeated application of undiluted isopro OcularIrritation pyl myristate for 21 days produced only slight irritation. Iso Previous safety assessments cited Draize testing of myristyl propyl myristate was not a human skin sensitizer when in myristate and isopropyl myristate at concentrations up to petrolatur or in product formulations at 15% to 58%, although 100% that produced minimum eye irritation in rabbits. Butyl a case report of sensitization was found. A product containing myristate (no concentration provided) was considered nonirri 43% isopropyl myristate produced no phototoxicity and no tating to the rabbit eye. Undiluted isopropyl myristate produced photocontact allergenicity in human studies.

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Myristicacid. 2 In clinical primary and cumulative irritation • butyl myristate, studies, myristic acid at concentrations of 100% or 40% to • calcium myristate, 50% in mineral oil were nonirritating. Mild-to-intense • cetyl myristate, erythema in single insult occlusive patch tests, soap chamber • decyl myristate, tests, and 21-day cumulative irritation studies were produced • ethylhexyl myristate, by cosmetic product formulations containing 2% to 93% myr • ethyl myristate, istic acid, and were generally not related to the fatty acid con • glyceryl dimyristate, centrations in the formulations. The Expert Panel also • glyceryl isostearate/myristate, considered data from other fatty acids (oleic, lauric, pamitic, • glyceryl myristate, and stearic) due to the structural similarities among these • isobutyl myristate, ingredients. • isocetyl myristate, • isodecyl myristate, Dermal Sensitization • isopropyl myristate, Ethyl myristate. 107Kligman applied ethyl myristate for • isostearyl myristate, 5 alternate-day 48-hour periods on the volar side of the arm • isotridecyl myristate, of 25 participants after pretreatment for 24 hours with 2.5% • lauryl myristate, aqueous sodium lauryl sulfate under occlusion. Sodium lauryl • magnesium myristate, • methyl myristate, sulfate (5%-15%) was applied to the test site for 1 hour before the application of the challenge. There were no signs of sensi • myristyl myristate, tization for either the 48- or 72-hour challenge. It was not stated • octyldodecyl myristate, in the text, but according to the Research Institute for Fragrance • oleyl myristate, Materials (RIFM),’° ethyl myristate was tested at 12%. • potassium myristate, 8 • propylene glycol myristate, • sodium myristate, Testing ProvocativeSkin • tetradecyloctadecyl myristate, Isopropyl myristote. Uter et performed a 9al’° retroactive • tridecyl myristate, and study of dermatitis patients patch tested for sensitization to iso • zinc myristate. propyl myristate. Isopropyl myristate was tested in 20% petro latum using 8117 patients and 10% petrolatum using 4554 Most of the esters are used as skin conditioning agents in cos patients between January 1992 and December 2001. The higher metics, but other functions include the following: anticaking concentration had 43 doubtful reactions, 5 irritant reactions, agents, emulsion stabilizers, viscosity increasing agents, 6 reactions, and 2 reactions. The lower + ++/-b++ concentra surfactants----cleansing agents, surfactants—emulsifying tion had 9 doubtful reactions, 2 irritant reactions, 7 reactions, + agents, slip modifiers, fragrance ingredients, hair conditioning and I reaction. The authors concluded that ++/+++ isopropyl agents, binders, film formers, and opacifiing agents. myristate does not need to be tested for during routine patch Myristic acid is produced by the saponification and fractio tests. nation of animal or vegetable fats and oils followed by isolation of the acid fraction that is then hydrogenated. Case Reports Analytical methods include TLC, gas-liquid chromatogra Isopropylmyristate.Bharati and King”° reported a 64-year- phy, x-ray powder diffraction, GC, infrared spectrometry, old woman who presented with an eczematous rash from a HPLC, MS, gravimetry, and histochemical staining. commercial sunscreen. Patch testing of the European standard Component fatty acids of myristic acid include n-tetradeca series gave positive results for formaldehyde, quaternium- 15, noic acid, n-hexadecanoic acid, and n-dodecanoic acid. Myris imidazolidinyl urea, and diazolidinyl urea. A further patch test tic acid and other myristates may contain unsaponifiable of the ingredients in the sunscreen resulted in positive reactions material, and some grades may contain glyceryl monomyris for isohexadacane 10% alcohol and isopropyl myristate 10% tate. Impurities in glyceryl myristate include glycerol, digly alcohol. cerol, and free fatty acid. Other impurities include heavy metals and arsenic. Summary Isopropyl myristate is the most commonly used ingredient in this assessment and is used in over 1000 products at concentra This report addressed the safety of the following inorganic salts tions of 0.001% to 82%. and esters of various fatty alcohols of myristic acid, including: Myristic acid, aluminum myristate, aluminum myristates/ palmitates, butyl myristate, cetyl myristate, glyceryl myristate, • aluminum dimyristate, isobutyl myristate, isocetyl myristate, isodecyl myristate, iso • aluminum isostearates/myristates, decyl myristate, isopropyl myristate, lauryl myristate, magne • aluminum myristate, sium myristate, myristyl myristate, octyldodecyl myristate, • aluminum myristates/palmitates, potassium myristate, propylene glycol myristate, sodium

Supplement Panel Book 1 Page 205 Alkyl Esters Supplement Book 1

I82S InternationalJournal of Toxicology29(Supplement 3) myristate, and zinc myristate are also reported as used andlor Isopropyl myristate up to 100% was nonirritating, nonirritat have reported concentration of use. ing in cumulative skin irritation tests, nonphototoxic, and non No uses or use concentrations were reported for aluminum photoallergenic in humans. isostearates/myristate, calcium myristate, decyl myristate, ethyl myristate, ethylbexyl myristate, glyceryl dimyristate, gly ceryl isostearate/myristate, isobutyl myristate, isostearyl myris tate, isotridecyl myristate, methyl myristate, oleyl myristate, Discussion tetradecyloctadecyl myristate, and tridecyl myristate. The data on butyl myristate and the related salts and esters, Myristic acid is approved as a food reagent and additive. coupled with the data on the related chemicals (myristic acid, Butyl myristate is also used as a plasticizer, as a lubricant for myristyl myristate, and isopropyl myristate), are a sufficient textiles, and in paper stencils. basis for a safety assessment. The CIR Expert Panel believes The myristates are readily hydrolyzed to the corresponding that there is little toxicological and chemical difference alcohols and acids, which are then further metabolized. Butyl between myristic acid and any of its inorganic salts included myristate may be readily hydrolyzed in vivo to its correspond in this report. The salts are expected to dissociate in any prod ing acid and alcohol, which are then further metabolized. uct formulation, independent of whether the salt is aluminum, When isopropyl myristate was aerosolized, 85% of the calcium, magnesium, potassium, sodium, or zinc. For the var absorbed isopropyl myristate was eliminated in 24 hours, ious esters of fatty alcohols and myristic acid, the CIR Expert mainly as exhaled carbon dioxide; very little labeled material Panel considers that these fatty acid esters are subject to hydro reached any tissues other than the lungs in monkeys. lysis to from myristic acid and the component fatty alcohols. Myristic acid, butyl myristate, and isopropyl myristate It is the experience of the Panel in its review of fatty alcohols enhanced the dermal penetration of several drugs. of varying length of carbon chains that there is little difference The IC50 of methyl myristate for the inhibition of rat brain in toxicity. Accordingly, the available data were considered sup prostaglandin D synthase and swine brain prostaglandin 2D portive of the safety of the entire group as used in cosmetics. dehydrogenase was >200 l.Lmol/L. The Expert Panel recognized that use concentration data are The acute oral 50LD of butyl myristate was >8 g!kg for rats. not available for all ingredients in this group and that some The acute oral 50LD for isopropyl myristate was >16 mL/kg in ingredients in the group are not in current use. The Expert Panel rats and 49.7 mL/kg in mice. considered that the use concentrations for the ingredients that Acute dermal application of butyl myristate (2 g!kg)was non- are in use are not likely to be different from the use concentra toxic and nonirritating to rabbits. When 10 rabbits were treated tions for other myristates. Were those ingredients not in current with a single dermal dose of ethyl myristate (5 g/kg) resulted in use to be used in the future? The Panel expects that they would be the death of 2 over 7 days. The intraperitoneal and subcutaneous used in products and at concentrations similar to those reported. 50LD for isopropyl myristate exceeded 79.5 mL!kg in rats and The Expert Panel recognized that these ingredients can the intraperitoneal 50LD was >50.2 mL/kg in mice. enhance the penetration of other ingredients through the skin. No death occurred, and no evidence of systemic toxicity was The Panel cautioned that care should be taken in formulating found at necropsy when the rats were exposed to aerosolized cosmetic products that may contain these ingredients in com isopropyl myristate. bination with any ingredients whose safety was based on Myristic acid, isopropyl myristate, and myristyl myristate their lack of dermal absorption data, or when dermal absorp were minimally irritating to the eyes of rabbits. Butyl myristate tion was a concern. was nonirritating to the rabbit eye. A number of the ingredients in this report—cetyl myristate, Myristic acid was nonirritating in a single insult occlusive octyldodecyl myristate, and sodium myristate—have uses that patch test and slightly irritating in a repeat open patch test on rab include sprays. There are no data available on inhalation toxi bits. Butyl myristate was a moderate skin irritant in rabbits and city for these ingredients or the other ingredients in this assess guinea pigs. Isopropyl myristate and myristyl myristate were ment. The Expert Panel determined that there is sufficient minimally irritating in several formulations in rabbits and mice. inhalation toxicity data on isopropyl myristate in its assessment Isopropyl myristate was nonirritating when injected parent demonstrating no inhalation toxicity. In addition to the inhala erally in albino rabbits. tion toxicity data, the Panel determined that butyl myristate and Butyl myristate and myristyl myristate were nonsensitizing the salts and esters can be used safely in hair sprays, because to guinea pigs. the ingredient particle size is not respirable. The Panel reasoned Isopropyl myristate and myristyl myristate were comedo that the particle size of aerosol hair sprays ( 38 jim) and pump genic to rabbit ears. hair sprays (>80 jim) is large compared with respirable particu Isopropyl myristate tested negative in the Salmonella! late sizes (10 jim). Microsome test in strains TA1538, TA1537, TA1535, There are no data on the reproductive or developmental toxi TA100, and TA98, with and without activation. city of myristic acid or its component parts for the derivatives. In clinical primary and cumulative irritation studies, myris Based on structure—activityrelationships, the Expert Panel con tic acid was nonirritating. Isopropyl myristate can produce sidered that these chemicals had little potential for such toxicity slight irritation but is not a human sensitizer at 15% to 50%. when used as cosmetic ingredients.

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Isopropyl myristate was not genotoxic in the Ames assay. 5. Cosmetic Ingredient Review Cetearyl alcohol, cetyl alcohol, The Expert Panel determined this to be sufficient carcinogeni isostearyl alcohol, myristyl alcohol, and behenyl alcohol. city data for the related ingredients in this safety assessment. Washington, DC: Author; 2006. 6. Andersen FA. Annual review of cosmetic ingredient safety assessments—2002/2003. mtJ Toxicol. 2005;24(S2):89-98. Conclusion 7. Final report of the amended safety assessment of glyceryl laurate, The CIR Expert Panel fmds that myristic acid, aluminum dimyr glyceryl laurate SE, glyceryl laurate/oleate, glyceryl adipate, gly istate, aluminum isostearates/myristates, aluminum myristate, ceryl alginate, glyceryl arachidate, glyceryl arachidonate, glyceryl aluminum myristates/palmitates, butyl myristate, calcium myris behenate, glyceryl caprylate, glyceryl caprylate, clyceryl capry tate, cetyl myristate, decyl myristate, ethyl myristate, ethyihexyl late/caprate, glyceryl citrate/lactate/linoleate/oleate, glyceryl myristate, glyceryl dimyristate, glyceryl isostearate/myristate, cocoate, glyceryl collagenate, glyceryl erucate, glyceryl hydroge glyceryl myristate, isobutyl myristate, isocetyl myristate, isode nated rosinate, glyceryl hydrogenated soyate, glyceryl hydroxys cyl myristate, isopropyl myristate, isostearyl myristate, isotride tearate, glyceryl isopalmitate, glyceryl isostearate, glyceryl cyl myristate, lauryl myristate, magnesium myristate, methyl isostearate/myristate, glyceryl isostearates, glyceryl lanolate, gly myristate, myristyl myristate, octyldodecyl myristate, oleyl myr ceryl linoleate, glyceryl linoleate, glyceryl montanate, glyceryl istate, potassium myristate, propylene glycol myristate, sodium myristate, glyceryl isotridecanoate/stearate/adipate, glyceryl ole- myristate, tetradecyloctadecyl myristate, tridecyl myristate, and ate SE, glyceryl oleate/elaidate, glyceryl palmitate, glyceryl palmi zinc myristate are safe as cosmetic ingredients in the current prac tate/stearate, glyceryl palmitoleate, glyceryl pentadecanoate, tices of use and concentration. Were ingredients in this group not glyceryl polyacrylate, glyceryl rosinate, glyceryl sesqioleate, in current use to be used in the future? The expectation is that they glyceryl/sorbitol/oleate/hydroxystearate, glyceryl stearate/acet would be used in product categories and at concentrations compa ate, glyceryl stearate/maleate, glyceryl tallowate, glyceryl rable to others in the group. thiopropionate and glyceryl undecylenate. mt J Toxicol. 2004;23(suppl 2):55-94. Authors’ Note 8. Elder RL. Final report on the safety assessment of myristyl myr istate and isopropyl myristate. JAm Coil Toxicol. 1982;4:5-80. Unpublished sources cited in this report are available from the Direc 9. Elder RL. Final report on the safety assessment of Cetearyl Alco tor, Cosmetic Ingredient Review, 1101 17th Street, Suite 412, Washington, DC 20036, USA. hol, Cetyl Alcohol, Isostearyl Alcohol, Myristyl Alcohol, and Behenyl Alcohol. JAm Coil Toxicol. 1988;7(3):359-413. Declaration of Conflicting Interest 10. Lanigan S. Final report on the safety assessment of methyl alco hol. mtJ Toxicol. 2001;20(suppl l):57-85. No potential conflict of interest relevant to this article was reported. 11. Andersen FA. Annual Review of Cosmetic Ingredient Safety F Alan Andersen, PhD, and Lillian C. Becker are employed by the Assessments—2004/2005. mt Toxicoi. 2006;25(S2):73-78. Cosmetic Ingredient Review. J 12. Elder RL. Final report on the safety assessment of Stearyl Alco hol, Oleyl Alcohol, and Octyl Dodecanol. J Funding Am Coli Toxicol. 1985;4(5): 1-29. The articles in this supplement are sponsored by the Cosmetic 13. JohnsonW Sr. Final report on the safety assessmentof propylene Ingredient Review. The Cosmetic Ingredient Review Program is glycol (PG) dicaprylate,PG dicaprylate/dicaprate,PG dicocoate, financially supported by the Personal Care Products Council. PG dipelargonate,PG isostearate, PG laurate, PG myristate,PG oleate,PG oleate SE,PG dioleate,PG dicaprate,PG diisostearate, References and PG dilaurate. mtJ Toxicol. 1999;18(2):35-52. 1. Elder RL. Final report onthe safetyassessmentof butyl myristate. 14. GottschalckT, Bailey IF. International Cosmetic Ingredient Die JAm Coil Toxicol. I990;9(2):247-258. tionaly and Handbook 2. Washington: CTFA 2008. 2. Elder RL. Final report on the safety assessment of oleic acid, lau 15. International Cosmetic Ingredient Dictionary and Handbook. 12th nc acid, palmitic acid, myristic acid, and stearic acid. JAm Coil ed. Washington, DC: CTFA; 2008. Toxicol. 1987;6(3):321-401. 16. Food Chemicals Codex. 3rd ed. Washington, DC: National Acad 3. Cosmetic Ingredient Review. Safety assessment of glyceryl emy Press; 1981. dilaurate, glyceryl diarachidate, glyceryl dibehenate, glyceryl 17. Fassett DW, Irish DD. Industrial Hygiene and Toxicology. New dierucate, glyceryl dihydroxystearate, glyceryl diisopalmitate, York, NY: Interscience Publishers; 1963:2:(2). glyceryl diisostearate, glyceryl dilinoleate, glyceryl dimyristate, 18. Windholz M, Budavari S, Blumetti RF, Otterbein ES, eds. The glyceryl dioleate, glyceryl diricinoleate, glyceryldipalmitate, Merck Index. Rahway, NJ: Merck and Ca; 1983:10. glyceryl dipalmitoleate, glyceryl distearate, glyceryl palmitate 19. Weast RC. CRC Handbook of Chemistry and Physics. Boca lactate, glyceryl stearate citrate, glyceryl stearate lactate, and gly Raton, FL: CRC Press; 1982:63. ceryl stearate succinate. Washington, DC: Author; 2005. 20. Swern D, ed. Bailey Industrial Oil and Fat Products. New York, 4. Elder RL. Final report on the safety assessment of cetearyl NY: John Wiley; 1979:4(1). alcohol, cetyl alcohol, isostearyl alcohol, myristyl alcohol, and 21. Gattefossb. Data sheet. MOD Unpublished data submitted by the behenyl alcohol. JAm Coli Toxicol. 1988;7:359-413. Council, 2008.

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I84S Internationaljournal of Toxicology29(Supplement3)

22. CTFA. Myristates: summary of unpublished safety data. Unpub 45. Lefort D, Paquot C, Pourchez A. Gas chromatography and lipo lished data submitted by CTFA, 1979. chemistry. VI. Comparison of the methyl, propyl, and isopropyl 23. CTFA. CTFA cosmetic ingredient chemical description. Lauric esters of fatty acids by gas chromatography. Oleagineux. 1962; acid. Unpublished data submitted by CTFA, 1979. 17:629-630. 24. Estrin NF, Haynes CR, Whelan JM, eds. CTFA Compendium of 46. Lutz DA, Eddy CR, Hunter JJ. X-ray diffraction study of some nor Cosmetic ingredient Composition. Spec(fications/Spectra. mal alkyl esters of long-chain acids. Lipids. 1967;2(3):204-207. Washington, DC: CTFA; 1982. 47. Horwitz W, ed. Official Methods ofAnalysis of the Association of 25. Cosmetic, Toiletries and Fragrance Association. Cosmetic Official Analytical Chemists (AOAC). 13th ed. Washington, DC: ingredient chemical description: Myristic acid. Unpublished data AOAC; 1980: 18. submitted by CTFA, 1979. 48. EstrinNF, Haynes CR, Wielan JW. Cosmetic Ingredient Descrip 26. Chemline. Online database.Chemical Abstract Society, 1988. tions. CTFA Compedium of Cosmetic Ingredient Composition. 27. Lewis Sr RJ, ed. Hawley’s Condensed Chemical Dictionary. Washington, DC: CTFA; 1982. New York, NY: John Wiley; 1997:13. 49. Senzel A, ed. Newburger’s Manual of Cosmetic Analysis. 28. Nikko Chemicals Co, Ltd. Specification NIKKOL ICR-R (iso Washington DC: AOAC; 1977:2. cetyl myristate). Unpublished data submitted by the Council, 2007. 50. Cosmetic, Toiletries and Fragrance Association. CTFA cosmetic 29. British Pharmaceutical Codex. London, England: The Pharma ingredient chemical description. Myristic acid. Unpublished data ceutical Press; 1973. submitted by CTFA, 1979. 30. Windholz M, ed. The Merck Index. Rahway, NJ: Merck and Co; 51. Cosmetic, Toiletries and Fragrance Association. CTFA cosmetic 1976:9. ingredient chemical description. Palmitic acid. Unpublished data 31. Wade A, ed. Martindale: The Extra Pharmacopoeia. London, submitted by CTFA, 1979. England: Pharmaceutical Press; 1977:27. 52. Cosmetic, Toiletries and Fragrance Association. CTFA cosmetic 32. Nikko Chemicals Co, Ltd. Specification NIKKOL ODM-l00 ingredient chemical description. Oleic acid. Unpublished data (Octyldodecetyl myristate). Unpublished data submitted by the submitted by CTFA, 1979. Council, 2007. 53. Cosmetic, Toiletries and Fragrance Association. CTFA cosmetic 33. Nikko Chemicals Co, Ltd. Specification NIKKOL potassium myris ingredient chemical description. Stearic acid. Unpublished data tate MK-140. Unpublished data submitted by the Council, 2007. submitted by CTFA, 1979. 34. Danisco Ingredients. UV-spectra for glycerol monoesters. Unpub 54. Takatori T, Terazawa K, Nakano K,.Matsumiya H. Identification lished data submitted by CTFA, 1999. of l0-hydroxy-12-octadecenoic acid in adipocere. Forensic Sci 35. CTFA. Myristates: Summary of unpublished data. (3ISa). Internat. l983;23(2-3):1 17-122. Unpublished data submitted by CTFA, 1979. 55. Van de Vaart FJ, Hulshoff A, Indemans AWM. Analysis of 36. Hawley CC, ed. Condensed Chemical Dictionary. 9th ed. creams. V. Application of thin-layer chromatography. Parts I and New York, NY: Van Nostrand Reinhold Co; 1977:10. II. Pharm Weekblad Sci. 1983;5(3): 109-118. 37. Nikko Chemicals Co, Ltd. Manufacturing process NIKKOL 1CM- 56. The characterization of long-chain fatty acids and their den R (isocetyl myristate). Unpublished data submitted by the Coun vatrves by chromatography. In: Ciddings JC, Grushka E, cil, 2008. Cazes J, Brown PR, eds. Advances in Chromatography. New 38. Cosmetic, Toiletries and Fragrance Association. Cosmetic ingre York, NY: Marcel Dekker; 1980. dient chemical description. Isopropyl myristate. Unpublished data 57. Smith RM. Recent advances in the high-performance liquid chro submitted by CTFA, 1979. matography of fatty acids. J Pharm Biomed Anal. 1983;l(2): 39. Cosmetic, Toiletries and Fragrance Association. Cosmetic ingre 143-151. dient chemical description. Myristyl myristate. Unpublished data 58. Davis DV, Cooks RG. Direct characterization of nutmeg constitu submitted by CTFA, 1979. ents by mass spectrometry-mass spectrometry. J Agric Food 40. Personal Care Products Council. Concentration of use—potential Chem. l982;30(3):495-504. additions to the butyl myristate report. Unpublished data sub 59. Shimasaki H, Ueta N. Fractionation of the neutral lipids of rice- mitted by the Council, 2008. bran oil by centrifugal liquid chromatography. Agric Biol Chem. 41. Nikko Chemicals Co, Ltd. Manufacturing process NIKKOL 1983;47(2) :327-329. 0DM-lOU (Octyldocetyl myristate). Unpublished data submitted 60. Cyong 3, Okada H. Histochemical studies on fatty acid in by the Council, 2008. lymphocyte-mediated immune reaction. Immunology. 1976; 42. Nikko Chemicals Co, Ltd. Manufacturing process NIKKOL 30(5):763-767. potassium myristate MD- 140, 2008. 61. Bailey AV, Pittmann RA. Wide-line NMR spectra of some satu 43. Taiko Oil Chem. Col. Ltd. Manufacturing process MNK-40 rated and unsaturated long chain fatty acids. JAm Oil Chem Soc. (water, postassium cocoate and potassium myristate. Unpublished 1971;48(12):775-777. data submitted by the Council, 2008. 62. Arudi RI. Sutherland MW, Bielski BHJ. Purification of oleic acid 44. Hashimoto A, Hirotani A, Mukai K. Thin-layer chromatography and linoleic acid. J Lipid Res. 1983;24(4):485-488. of true wax 8. Nippon Nogei Kagaku Kaishi. l967;41(4): 63. Cotte M, Dumas P, Besnard M, Tchoreloff P, Walter P. Synchro 139-144. tron FT-IR microscopic study of chemical enhancers in

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acids.

Alanazi ML,

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evaluation

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2002;25(3):392-396.

the

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2008;3 Tech.

propylene

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melatonin acids

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2008;9(2):464-470.

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2008;31(9):1766-1772.

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1978;29:265-282.

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distribution,

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1):

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skin

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transport

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2005;60(3):215-220.

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In a Alkyl Esters Supplement Book 1

I86S InternationalJournal of Toxicology29(Supplement 3)

99. Laboratoire de Recherche et d’Experimentation. Attestation 105. Nguyen SH, Dang TP, Maibach HI. Comedogenicity in rabbit: of biological test. Unpublished data submitted by CTFA, some cosmetic ingredients/vehicles. Cutaneous Ocular Toxicol. 1994:. 2007;26:287-292. 100. Food and Drug Research Laboratories Inc. Report to RIFM. 5- 106. Blevins RD, Taylor DE. Mutagenicity screening of twenty-five 2 1-1976. cosmetic ingredients with the Salmonella/microsome test. J

101. CTFA. 1978. Acute Dermal Toxicity in Rabbits. (31.5b. p. 35). Environ Sci Health Part A. 1982;17:(2):2l4-239. Unpublished data submitted by CTFA. 107. Kligman AiM.Report to RIFM dated June 1, 1976. Unpublished 102. Motoyoshi K. Enhanced comedo formation in rabbit ear skin by data submitted by RIFM, 1976:2 pages. sqalene and oleic acid peroxides. Br J Dermatol. 1983;109: 108. Research Institute for Fragrance Materials. Ethyl myristate. 191-198. Food and Chem Toxicol. l978;l6:(4):745-746. 103. Fulton JE, Pay SR. Comedogenicity of current therapeiutic prod 109. Uter W, Schnuch A, Heier J, Lessmann H. Isopropyl myristate ucts, cosmetics, and ingredients in the rabbit ear. JAm Acad Der recommended for aimed rather than routine patch testing. Con matol. l984;lO:96-105. tact Dermatitis. 2004;50:242-244. 104. Tucker SB, Flanigan SA, Dunbar M, Drotman RB. Development 110. Bharati A, King CM. Allergic contact dermatitis from isohexa of an objective comedogenicity assay. Arch Dermatol. 1986; decane and isopropyl myristate. Contact Dermatitis. 2004; 122:660-665. 50(256):257.

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Final Report of the Cosmetic Ingredient Review Expert Panel

On the Safety Assessment of Pelargonic Acid (aka Nonanoic Acid) and Nonanoate Esters

June 29, 2010

The 2010 Cosmetic Ingredient Review Expert Panel members are: Chairman, Wilma F. Bergfeld, M.D., F.A.C.P.; Donald V. Belsito, M.D.; Ronald A. Hill, Ph.D; Curtis D. Klaassen, Ph.D.; Daniel Liebler, Ph.D; James G. Marks, Jr., M.D., Ronald C. Shank, Ph.D.; Thomas J. Slaga, Ph.D.; and Paul W. Snyder, D.V.M., Ph.D. The CIR Director is F. Alan Andersen, Ph.D. This report was prepared by Wilbur Johnson, Jr., Senior Scientific Analyst/Writer and Bart Heldreth, Ph.D., Chemist.

Co s m e t i c I n g r e d i e n t R e v i e w 1101 17th Street, NW, Suite 412 ♢ Washington, DC 20036-4702 ♢ (202) 331-0651 ♢ [email protected]

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1101 17th Street, NW, Suite 412 Washington, DC 20036-4702

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ABSTRACT

Pelargonic Acid and its esters function as skin conditioning agents in cosmetics. Molecular weight and octanol-water partition coefficient data suggest that dermal penetration is possible. The biohandling of branched-chain fatty acids is not the same as for straight-chain fatty acids, but the differences are not significant to the conclusion that they all are readily metabolized to non-toxic moieties. Limited data suggested that the penetration of other ingredients may be enhanced if these ingredients are present in the same formulation. These ingredients are not significant oral or dermal toxicants in animal studies. They are not reproductive/developmental toxicants or genotoxic/carcinogenic in animal studies. The available data suggested that product formulations containing these ingredients would be nonirritating and non-sensitizing to human skin, but formulators were cautioned to consider the penetration enhancement potential. The CIR Expert Panel concluded that these ingredients are safe in the present practices of use and concentration.

INTRODUCTION

Pelargonic acid (aka nonanoic acid) is a fatty acid that can function as a fragrance ingredient, surfactant- cleansing agent, and surfactant-emulsifying agent in cosmetics. In its soap form, it can function as a surfactant- cleansing agent. Many of the fatty acids that are used in cosmetics and their synthesized derivatives (primarily esters and diesters of the corresponding alcohol and pelargonic acid) have similar additional functions in cosmetics. An idiosyncrasy in the terminology used in the International Cosmetic Ingredient Dictionary and Handbook is that most of these derivatives are termed nonanoates, not pelargonates. This safety assessment includes the following ingredients:

 pelargonic acid  butylene glycol diisononanoate  cellobiose octanonanoate  cetearyl isononanoate  cetearyl nonanoate  cetyl isononanoate  cholesteryl nonanoate  diethylene glycol diethylhexanoate/diisononanoate  diethylene glycol diisononanoate  dihydrocholesteryl nonanoate  dipentaerythrityl pentaisononanoate  ethylhexyl isononanoate  glycereth-7 diisononanoate  isodecyl isononanoate  isononyl isononanoate  isostearyl isononanoate  isotridecyl isononanoate  neopentyl glycol diisononanoate  PEG-2 diisononanoate  PEG-5 isononanoate  pentaerythrityl tetraisononanoate  phytosteryl nonanoate  polyglyceryl-20 octaisononanoate  propylene glycol diisononanoate  tridecyl isononanoate  ethylhexyl pelargonate  ethyl pelargonate  isobutyl pelargonate 1

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 methyl pelargonate  neopentyl glycol dicaprylate/dipelargonate/dicaprate  pentaerythrityl tetrapelargonate

The nonanoate or pelargonate esters and diesters are straight-chain compounds, whereas the isononanoate esters and diesters are branched-chain compounds. Propylene glycol dipelargonate would have been included in this safety assessment; however, the Cosmetic Ingredient Review (CIR) Expert Panel previously concluded that this ingredient is safe in the present practices of use in cosmetic products. 1

Pelargonic acid is a reactant in the esterification process that yields all of the esters included in this safety assessment. While this fatty acid is not reported to be currently used in cosmetics, much of the data available for review relate to pelargonic acid. The CIR Expert Panel has published safety assessments on the following fatty alcohols and other reactants used to form some of the esters reviewed in this safety assessment, and other fatty acid esters: butylene glycol - safe in present practices of use and concentration [safe],2,3 cetearyl, cetyl, and isostearyl alcohols – safe,4,5 cholesterol – safe,6,3 ethylene glycol – special report on reproductive and developmental toxicity - no conclusion,7 ethylhexyl palmitate (previously incorrectly named octyl palmitate) – safe,8,9 PEG-7 glyceryl cocoate – safe as used in rinse-off products; safe up to 10% in leave-on products,10 PEGs-2 and -6 dilaurate – safe for use in cosmetics at concentrations up to 25%,11 octyl stearate and isobutyl stearate – safe, 12,5 isodecyl oleate – safe,13,14,14 and propylene glycol dipelargonate – safe,1 as stated above. These conclusions may contribute to the safety assessment of ingredients in the current review for which little or no data have been identified in the published literature.

Similarly, there are ingredient moieties that have not been reviewed by the Expert Panel, and available data on these chemicals may be useful in the absence of safety test data on some of the esters that are being reviewed. Thus, data on or relevant to the following chemicals are also summarized in the current safety assessment: isononanoic acid, isononyl alcohol, isotridecyl alcohol, neopentyl glycol, isobutyl alcohol, and isodecyl alcohol. Excerpts from the summary and discussion from the CIR Final Safety Assessment on propylene glycol dipelargonate and other propylene glycol esters and diesters are also included, because these data may be useful in evaluating the safety of diesters included in the current review. Similarly, excerpts from the CIR Final Safety Assessments on isobutyl stearate and isodecyl oleate are also included, in lieu of data on isobutyl alcohol and isodecyl alcohol, respectively.

CHEMISTRY

DEFINITION AND STRUCTURE Chemical definitions, other chemical names, and cosmetic ingredient functions for the ingredients reviewed in the safety assessment are included in Table 1. The International Nomenclature Cosmetic Ingredient (INCI) name appears first in each series of chemical names; “iso” in an INCI name denotes methyl branching/substitution and does not necessarily imply substitution on the second carbon atom.15 Chemical structures/formulas are included in Figure 1. The inclusion of [sic] after a technical name or CAS No. in Table 1 denotes those instances wherein the authors of the dictionary associated a specific branched chemical entity other than an omega-2 methyl substituted isomer with an “iso” INCI name. According to the dictionary proviso regarding “iso”-named ingredients, all branched isomers are potentially included by an “iso” INCI ingredient name. For simplicity, only omega-2 isomers are shown in Figure 1. However, in one exception, the [sic] notation is included after “octyl isononanoate,” in the ethylhexyl isononanoate box of Table 1, in that octyl isononanoate is included in the International Cosmetic Ingredient Dictionary and Handbook because it is a former INCI name for this ingredient.

CHEMICAL AND PHYSICAL PROPERTIES Available chemical and physical properties are included in Table 2.

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UV ABSORPTION Data provided by Symrise GmbH & Co.16 indicated that the following chemicals did not absorb significantly in the 250 to 400 nm range: neopentyl glycol diisononanoate, cetyl nonanoate + stearyl nonanoate, trideceth-9 + PEG- 5-isononanoate + water, glyceryl triisononanoate + glyceryl diisononanoate, and ethylhexyl isononanoate.

ANALYTICAL METHODS Pelargonic acid, cholesteryl nonanoate, ethyl pelargonate, and methyl pelargonate have been analyzed by nuclear magnetic resonance (NMR), infrared (IR), and mass spectrometry, and pentaerythrityl pelargonate has been analyzed by IR spectrometry .17 Methyl pelargonate and pelargonic acid have also been analyzed by gas chromatography-mass spectrometry,18 and the same is true for ethyl pelargonate .19

METHODS OF PRODUCTION In general, the alkyl esters can be produced industrially via esterification of carboxylic acids with the corresponding alcohols (with or without a metal catalyst).20 The source of these carboxylic acids and alcohols is often natural, or is simply derivied from natural sources. These sources are often mixtures. This is especially true in the case of branched acids and alcohols. Accordingly, the resulting esters are also mixtures. Although many carboxylic acids and alcohols are obtained from natural sources, alcohols with chains longer than ethanol (C2) are sometimes produced synthetically. An important process for producing C3- C20 industrial alcohols involves a process known as oxo- synthesis (a process for the production of aldehydes which occurs by the reaction of olefins (which can be natural or petroleum sourced) with carbon monoxide, hydrogen and a catalyst (typically colbalt based)), followed by hydrogenation of the aldehyde products, to form the alcohols.21 An industry shift began a couple of years ago towards a green, biocatalytic process developed specifically for the manufacture of esters for use in the formulation of cosmetic and personal care ingredients (i.e. for producing cosmetic grade esters).22

Pelargonic acid is prepared from unsaturated hydrocarbons by the oxo process, or by the oxidation of oleic acid, and from tall oil unsaturated fatty acids or rice bran oil fatty acid.23 In the oxo process, pelargonic acid is prepared synthetically. Preparation from tall oil unsaturated fatty acids or rice bran oil fatty acid occurs naturally via splitting/separation.

The production methodology for cetearyl isononanoate and isononyl isononanoate includes use of a typical mineral salt/non-organic catalyst for ester formation in the reaction; the catalyst is removed upon completion of the reaction. 24 Actually, the catalyst is filtered off.

IMPURITIES

Accordingly, typical impurities include olefin, acid, and alcohol starting materials; water; and residual metals (from catalysts).

Specifications on pelargonic acid from a chemical supplier include pelargonic acid (90% minimum), iron (1.0 ppm max), moisture (0.2% max), and the following monobasic acids: C9 (93%), and other monobasic acids (2%).25 The C9 acid (i.e., isononanoic acid) and other monbasic acids (e.g., octanoic acid) are also referred to as monoprotic acids.

The heavy metals content of both cetearyl isononanoate (Tegosoft CI) and isononyl isononanoate (Tegosoft INI) is described as follows: 20 ppm maximum (copper; lead; tin; platinum; palladium; mercury; arsenic; cadmium; nickel); < 1 ppm (mercury; arsenic; cadmium; nickel).26, 27

Neopentyl glycol formic acid ester and neopentyl glycol isolactic acid ester are impurities that have been detected in neopentyl glycol. 28

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USE

PURPOSE IN COSMETICS

The majority of the ingredients reviewed in this safety assessment function as skin conditioning agents in cosmetics. Cholesteryl nonanoate is the only ingredient for which an ingredient function in cosmetics is not listed in the International Cosmetic Ingredient Dictionary and Handbook. Ingredient functions in cosmetics are included in Table 1.29

SCOPE AND EXTENT OF USE IN COSMETICS

According to information supplied to the Food and Drug Administration (FDA) by industry as part of the Voluntary Cosmetic Registration Program (VCRP) in 2009,30 only the following ingredients reviewed in this safety assessment are being used in cosmetic products: cetearyl isononanoate, cholesteryl nonanoate, diethylene glycol diethylhexanoate/diisononanoate, ethylhexyl isononanoate, isodecyl isononanoate, isononyl isononanoate, isotridecyl isononanoate, tridecyl isononanoate, and ethylhexyl pelargonate. These use data are summarized in Table 3. Independent of these data, the results of a survey of current ingredient use concentrations that was conducted by the Personal Care Products Council in 2009 are also summarized in Table 3.31 For example, cetearyl isononanoate is used in 5 of the 1196 eye shadow products reported to the VCRP, and results from the separate industry survey indicate use of this ingredient at a concentration of 0.05% in these products. In other cases, for example, uses are reported to the VCRP, but use concentration data are not available, as for cetearyl isononanoate. Current use concentration data from the Personal Care Products Council also indicate that, while not reported to the VCRP, the following ingredients are also being used in cosmetic products: butylene glycol diisononanoate, cetearyl nonanoate, cetyl isononanoate, dipentaerythrityl pentaisononanoate, neopentyl glycol diisononanoate, PEG-2 diisononanoate, PEG-5 isononanoate, pentaerythrityl tetraisononanaote, polyglyceryl-20 octaisononanoate, and pentaerythrityl tetrapelargonate, as shown in Table 3.

Based on the data included in Table 3 (use frequency and use concentration data), there is no indication that the following ingredients are being used in cosmetics: cellobiose octanonanoate, diethylene glycol diisononanoate, dihydrocholesteryl nonanoate, glycereth-7 diisononanoate, isostearyl isononanoate, phytosteryl nonanoate, propylene glycol diisononanoate, ethyl pelargonate, isobutyl pelargonate, methyl pelargonate, neopentyl glycol dicaprylate/dipelargonate/dicaprate, and pelargonic acid.

Cosmetic products containing the in-use ingredients may be applied to the skin or hair, or, incidentally, may come in contact with the eyes, nails, and mucous membranes. Products containing these ingredients may be applied as frequently as several times per day and may remain in contact with the skin/hair for variable periods following application. Daily or occasional use may extend over many years.

NONCOSMETIC USE

Pelargonic acid is included on the list of food additives (synthetic flavoring substances and adjuvants) permitted for direct addition to food for human consumption, as stated in 21 CFR 172.515.32 According to 21 CFR 173.315,32 an aliphatic acid mixture consisting of valeric, caproic, enanthic, caprylic, and pelargonic acids is approved for use at a level not to exceed 1% in a lye peeling solution used in the peeling of fruits and vegetables (21 CFR 173.315).32 Another approved use of pelargonic acid relates to sanitizing solutions that may be used safely on food- processing equipment and utensils, and on other food-contact articles. One such solution contains decanoic acid, pelargonic acid, phosphoric acid, propionic acid, and sodium 1-octanesulfonate, according to 21CFR 178.1010.32

Pelargonic acid is registered by the United States Environmental Protection Agency (EPA) for use as a blossom thinner and as an herbicide. 33,34 It is exempt from the requirement of a tolerance for pesticide residues in or on all foods, when used as a component of a food contact surface sanitizing solution in food handling establishments.35

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GENERAL BIOLOGY

METABOLISM

Straight-chain pelargonic acid esters are likely hydrolyzed to component alcohols and pelargonic acid, which is further metabolized to acetate moieties that enter the citric acid cycle. Branched-chain fatty acid metabolism involves initial α-oxidation, which is followed by the β-oxidation pathway.

Pelargonic Acid

The oxidative degradation of fatty acids, such as pelargonic acid, into 2-carbon fragments through enzymatically-catalyzed reactions is a well-documented central metabolic pathway in animals and plants. Pelargonic acid, a straight-chain carbon molecule, would be metabolized by beta-oxidation to form acetate molecules, which enter the citric acid cycle and are metabolized to carbon dioxide, water, and energy.33,36

Ethyl Pelargonate According to Opdyke,37 straight-chain aliphatic acid esters are thought to be readily hydrolyzed into their component acids and alcohols, which would then be expected to follow their normal metabolic pathways.

Branched-Chain Fatty Acids Mammalian metabolism of some lipids, including 3-methyl (e.g., phytanic acid) and 2-methyl (e.g., pristanic acid) branched-chain fatty acids occurs in peroxisomes.38 Because of the location of a methyl group at the β-carbon of phytanic acid, degradaion of the acid via the β-oxidation pathway cannot occur. Instead, the α-methylene group of phytanic acid is is oxidatively excised, yielding pristanic acid, which can be metabolized via the β-oxidation pathway.

PERCUTANEOUS ABSORPTION Of the ingredients included in Table 2, most have a log P of >5 and a mw of < 500. The exceptions are as follows: cellobiose octanonanoate,cholesteryl nonanoate, and pentaerythrityl tetrapelargonate (log P >5; mw > 500); ethyl pelargonate and methyl pelargonate (logP < 5; mw < 500); and isobutyl pelargonate (log P > 5; mw < 500) Compounds with molecular weights >500 and logP values > 5 are less likely to penetrate the skin. Actual skin penetration data (rabbits, abraded skin) indicate that isononyl alcohol, under occlusion, is percutaneously absorbed.

Isononyl Alcohol The results of an acute dermal toxicity study39 on undiluted isononyl alcohol (rabbits, abraded skin) are summarized later in the report text. When the occlusive binders were observed after 24 h of contact in this study, percutaneous absorption was evident. Details relating to this finding were not included.

SKIN PENETRATION ENHANCEMENT

The skin penetration enhancement effect of pelargonic acid on other chemicals has been demonstrated in vitro. Pelargonic acid enhanced the skin penetration of PABA through human stratum corneum and, melatonin, through hairless rat skin.

Tanojo and Junginger40 studied the skin penetration enhancement effects of fatty acids on p-aminobenzoic acid (PABA) penetration through sheets of human stratum corneum (surgically removed human breast or abdominal skin). The stratum corneum sheet was pre-treated with the penetration enhancer. The permeation of PABA increased with increasing chain-length of straight-chain saturated fatty acid, from 6 to 9 carbons (hexanoic, heptanoic, octanoic, and pelargonic acids, respectively). A sharp increase in PABA permeability occurred at fatty acid chain lengths between 8 and 9 carbons. The mean steady state flux for PABA was 837.84 ± 190.30 µg/cm2/h in the presence of pelargonic acid, compared to 2.57 ± 0.19 µg/cm2/h in the presence of water and 0.28 ± 0.14 µg/cm2/h in the presence of propylene glycol. 5

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Kanikkannan et al.41 conducted a study on the release profile of melatonin from drug-in-adhesive type transdermal patches (prepared using Eudragit® E 100 as adhesive polymer) containing pelargonic acid or other penetration enhancers, and the in vitro penetration of melatonin through hairless rat skin in the presence of the enhancer. Melatonin release was studied using the United States Pharmacopoeia dissolution test apparatus in conjunction with high performance liquid chromatography. The release profiles of melatonin from the patches with enhancers were similar when compared to the control patch release profile.

In skin penetration experiments, each penetration enhancer was added to the patch at a concentration of 2.5% or 5%. Skin samples from at least 3 rats were used in each experiment, and each mean value for skin penetration represented 3 replicates. The presence of enhancers in the patches resulted in an increase in the permeation of melatonin through hairless rat skin. The mean melatonin flux values for patches containing octanol, pelargonic acid, or myristic acid (each at both concentrations) were higher when compared to the control patch; however, the differences were not statistically significant (P > 0.05). Each mean value represented 3 replicates.41

ENDOCRINE DISRUPTION

According to EPA,36 it would be unlikely for straight chain carbon molecules, as in the C9 carbon chain of pelargonic acid, to be associated with a risk of endocrine disruption.

ANIMAL TOXICOLOGY

ACUTE INHALATION TOXICITY

An acute inhalation LC50 of 1.34 mg/L was reported for pelargonic acid in a study involving rats. Inhalation exposure to isononanoic acid caused a concentration- dependent decrease in respiratory frequency in mice, and an RD50 of 420 mg/m3 was reported. Few and no deaths were reported for mice/rats and guinea pigs, respectively, following inhalation exposure to isononyl alcohol at a concentration of 21.7 mg/L.

Pelargonic Acid EPA33 placed pelargonic acid in toxicity category III (> 0.5 through 2 mg/liter), primarily based on the results of the following study. The acute inhalation toxicity of pelargonic acid was evaluated using groups of 10 (5 males, 5 females/group) albino rats .42 The 4 groups were exposed (4-hour exposure) to aerosol generated from undiluted pelargonic acid, delivering concentrations of 0.510, 0.710, 2.20, and 3.31 mg/L, respectively. The following mortalities were reported: 1 rat (at 0.510 mg/L), 1 rat (at 0.710 mg/L), 8 rats (at 2.20 mg/L), and 10 rats (at 3.31 mg/L). Gross necropsy was performed on animals that died, and the following findings were considered unusual and possibly related to exposure: nasal discharge; polyuria; salivation; and discolored and swollen lungs, and variations thereof. Acute inhalation LC50’s of 0.87 mg/L (95% confidence limits = 0.50 to 1.51 mg/L) and 2.10 mg/L (95% confidence limits = 1.71 to 2.58 mg/L) were reported for males and females respectively. The overall LC50 was 1.24 mg/L (95% confidence limits undefined).

Isononanoic Acid Detwiler-Okabayashi43 evaluated the respiratory effects of isononanoic acid using groups of 4 specific pathogen-free, male Swiss-Webster mice. The animals were exposed to nebulized isononanoic acid (concentration range: 172 to 755 mg/m3) in a 2.5 l exposure chamber for 180 min. Sensory and pulmonary irritation were reported, and recovery immediately post-exposure was described as poor. The test substance concentration that was capable of evoking a 50% decrease in the mean respiratory frequency (RD50) was 420 mg/m3. The decreases in respiratory frequency induced by isononanoic acid were described as concentration-dependent and due to a combination of sensory and pulmonary irritation.

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Isononyl Alcohol Scala and Burtis39 studied the inhalation toxicity of isononyl alcohol using groups of 10 Swiss mice, Wistar rats, and English Short Hair guinea pigs. Each group received a single 6-h vapor exposure under dynamic conditions; exposure was followed by a 14-day observation period. The concentration of isononyl alcohol in the exposure chamber was calculated to be 21.7 mg/liter. None of the animals died during exposure; however, one mouse and 2 rats died within the first 14 h post-exposure. Signs of systemic toxicity consisted primarily of central nervous system depression, but were not pronounced. Local irritation involving mucous membranes of the eyes and nose was observed to a variable extent, and all animals had recovered within 1 h after termination of exposure. Histopathological examinations were not performed.

ACUTE ORAL TOXICITY

Pelargonic acid and the esters for which data are available are not significant acute oral toxicants (LD50s > 1 g/kg).

Pelargonic Acid EPA33 placed pelargonic acid in toxicity category IV (> 5000 mg/kg) , primarily based on the results of the following study. The acute oral toxicity of nonanoic acid was evaluated using 2 groups of 10 specific-pathogen-free SD (Crj:CD) rats (5 males, 5 females/group).44 One group was dosed orally with nonanoic acid in corn oil (dose = 5,000 mg/kg). The control group was dosed with vehicle only. None of the animals died and no abnormal clinical signs were noted during the 14-day observation period. There was no evidence of macroscopic abnormalities at necropsy. The LD50 was > 5,000 mg/kg, and nonanoic acid was considered nontoxic.

Cetearyl Nonanoate The acute oral toxicity of cetearyl nonanoate (97% pure) was evaluated using groups of 4 Sprague-Dawley CD rats.45 One group was dosed orally with 300 mg/kg and the remaining 2 groups were dosed with 2,000 mg/kg. None of the animals died. Signs of systemic toxicity were observed in the 300 mg/kg dose group, but not in the 2,000 mg/kg dose group. Necropsy did not reveal any abnormal findings, and an LD50 of > 2,000 mg/kg body weight was reported.

Cetearyl Isononanoate According to Evonik Industries,46 an LD50 of > 5,000 mg/kg was reported for cetearyl isononanoate in a study involving mice. Study details were not provided.

Isononyl Isononanoate According to Evonik Industries ,47 an LD50 of > 5,000 mg/kg was reported for isononyl isononanoate in a study involving rats. Study details were not provided.

Ethyl Pelargonate According to Opdyke,37 acute oral LD50 values of > 43,000 mg/kg and 24,190 mg/kg have been reported for rats and guinea pigs, respectively.

Ethylhexyl Pelargonate The acute oral toxicity of undiluted ethylhexyl pelargonate was evaluated using 10 albino rats (5 males, 5 females).48 A single oral dose of 5 g/kg body weight was administered to each animal. Dosing was followed by a 14- day observation period, and gross necropsy was performed on animals that survived. The LD50 was > 5 g/kg.

Isononyl Alcohol Scala and Burtis39 evaluated the acute oral toxicity of isononyl alcohol using 5 fasted, male Sprague-Dawley rats. An acute oral LD50 of 2.98 g/kg was reported.

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Isotridecyl Alcohol According to Guilian and Naibin,49 the acute oral LD50 for isotridecyl alcohol in rats is 17,000 mg/kg.

Neopentyl Glycol Diisononanoate The acute oral toxicity of undiluted neopentyl glycol diisononanoate was evaluated using groups of 4 Sprague-Dawley CD rats.50 One group was dosed orally with 300 mg/kg and the remaining 2 groups were dosed with 2,000 mg/kg. None of the animals died, and there were no signs of systemic toxicity in either of the 3 groups. Necropsy did not reveal any abnormal findings, and an LD50 of > 2,000 mg/kg body weight was reported.

PEG-5 Isononanoate The acute oral toxicity of PEG-5 Isononanoate was evaluated using 2 groups of 3 fasted Sprague-Dawley CD rats.51 Animals of both groups were dosed orally with 2,000 mg/kg. None of the animals died and necropsy did not reveal any abnormal findings. An LD50 of > 2,500 mg/kg body weight was reported.

Neopentyl Glycol According to Guilian and Naibin,49 the acute oral LD50 for neopentyl glycol in rats is 3259 mg/kg. The Organisation for Economic Co-operation and Development (OECD)28 reported an acute oral LD50 of 3200 mg/kg (rats) for neopentyl glycol.

ACUTE DERMAL TOXICITY

Pelargonic acid and the esters for which data are available are not significant acute dermal toxicants (LD50s of >2,000 to 9,000 g/kg).

Pelargonic Acid In a study involving rabbits,37 the acute dermal LD50 was greater than 5 g/kg (number of animals not stated). Results relating to the skin irritation potential of pelargonic acid in this study are included later in the report text. EPA33 placed pelargonic acid in toxicity category III (> 2000 through 5000 mg/kg), based on the results of the following acute dermal toxicity studies that were published in a Federal Register notice:36 The application of pelargonic acid to intact and abraded skin of mice induced moderate to severe skin irritation, and an acute dermal LD50 of 9000 mg/kg was reported in this study. An acute dermal LD50 of 5000 mg/kg (rabbits) for undiluted pelargonic acid also has been reported.

The acute dermal toxicity of nonanoic acid was evaluated using 2 groups of 10 specific-pathogen-free SD (Crj:CD) rats (5 males, 5 females/group) .52 The test substance, in deionized water, was placed on filter paper that was applied to clipped, shaved skin (4 x 5 cm site) of the back for 24 hours. Deionized water (0.5 ml) was applied to control animals according to the same procedure. None of the animals died during the 21-day observation period. Scales/scabs in the dorsal region (test substance-related) were observed only in treated females (days 3 to 17 post-application). Macroscopic abnormalities were not observed in any of the animals (test or controls) at necropsy. It was concluded that the LD50 for nonanoic acid in males and females was > 2,000 mg/kg.

Cetearyl Nonanoate The acute dermal toxicity of cetearyl nonanoate (97% pure) was evaluated using 10 Sprague-Dawley CD strain rats (5 males, 5 females).45 The test substance was applied to intact skin (24 h semi-occlusive application) at a dose of 2,000 mg/kg body weight. None of the animals died and there were no signs of systemic toxicity or dermal irritation. Necropsy findings were not indicative of any abnormalities, and an LD50 of > 2,000 mg/kg body weight was reported.

Ethyl Pelargonate According to Opdyke,37 the acute dermal LD50 in rabbits exceeded 5 g/kg.

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Isononyl Alcohol Scala and Burtis39 evaluated the acute dermal toxicity of undiluted isononyl alcohol using 4 rabbits. The test substance was applied (under occlusive binding) to clipped, abraded abdominal skin at the following doses: 0.500, 0.200, 0.794, and 3.16 g/kg. An acute dermal LD50 of 3.2 g/kg was reported. Signs of percutaneous toxicity were not observed. Limited results relating to the percutaneous absorption of isononyl alcohol are included earlier in the report text. Results relating to the skin irritation potential of isononyl alcohol are included later in the report text.

Neopentyl Glycol Diisononanoate The acute dermal toxicity of undiluted neopentyl glycol diisononanoate was evaluated using 10 Sprague- Dawley CD strain rats (5 males, 5 females).50 A dose of 2,000 mg/kg body was applied according to the procedure in the preceding section on Cetearyl Nonanoate. None of the animals died and there were no signs of systemic toxicity or dermal irritation. Necropsy findings were not indicative of any abnormalities, and an LD50 of > 2,000 mg/kg was reported.

ACUTE INTRAVENOUS TOXICITY

Oro and Wretlind 53 studied the acute intravenous (i.v.) toxicity of pelargonic acid using 10 mice (weights = 13 to 34 g). An LD50 of 224 ± 6 mg/kg was reported. Similarly, an LD50 of 224 mg/kg was reported for mice dosed i.v. with undiluted pelargonic acid.36

ACUTE INTRAPERITONEAL TOXICITY

In a study involving rats, intraperitoneal (i.p.) dosing with undiluted pelargonic acid resulted in death, and the lowest lethal dose (LDLo) was 3200 mg/kg.36 The dosing (i.p.) of mice with a 10% solution of pelargonic acid in corn oil resulted in death, and an LDLo of 1600 mg/kg was reported.36

SHORT-TERM ORAL TOXICITY

Pelargonic Acid A study by Hazleton Labs America Inc.54 was conducted to determine the appropriate dose level of pelargonic acid for a teratology screening study. Groups of 6 Cesarean-derived, Sprague-Dawley rats (sexually mature; weights = 177 to 285 g) were used. The test substance was administered (via oral intubation) in corn oil, at 3 dose levels (200, 1000, and 2000 mg/kg/day; 6 inseminated females/dose level) from days 6 to 15 of gestation. The dose volumes corresponding to the administered doses (lowest to highest) were 1.0, 5.0, and 10.0 ml/kg. The control group (6 inseminated females) was dosed with corn oil according to the same procedure. All surviving rats were killed after gestation day 15 and necropsied.

There were no remarkable clinical signs in any of the rats dosed with pelargonic acid, and none of the rats died. Mean feed consumption in the 200 mg/kg dose group was significantly higher, up to gestation day 14, when compared to the control group. Gross pathology findings were observed principally in the lungs, kidneys, or stomach. The numbers of rats with gross lesions included 2 rats at the 200 mg/kg dose level and one rat each at the 1000 and 2000 mg/kg levels. It was agreed that the only pelargonic acid-induced effect was on body weight. Study results relating to reproductive and developmental toxicity are included in the Section on Reproductive and Developmental Toxicity later in the report text.54

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In another short-term study,55 the oral toxicity of pelargonic acid was evaluated using groups of 6-week-old albino rats (Crl:CD®(SD)BR strain). Four groups (15 males and 15 females per group) were fed pelargonic acid in the diet at concentrations of 10, 100, 1000, and 5000 ppm, respectively. Ten rats per sex in each group received their respective diet for 28 days and until necropsy; a fifth group received basal diet only. After 28 days of pelargonic acid (in diet) feeding, the remaining rats (5 males and 5 females per group) in the 4 groups were switched to a basal diet, and feeding was continued for an additional 56 days (recovery phase). The fifth group was allowed to continue on the basal diet. Necropsy was also performed at the end of the recovery phase. The results of ophthalmic examinations are included in the section entitled Ocular Irritation/Toxicity later in the report text. Compared to the control group, male rats in the 5000 ppm dietary group had significantly lower (p < 0.05) group mean body weights during weeks 1 through 4. The same was true for female rats in this group during weeks 3 through 6. Changes in mean body weights, body weight gains, and food consumption in the 100 ppm and 1000 ppm dietary groups were influenced by an approximately 30-h water deprivation, which occurred during week 3. Reversible changes in clinical pathology variables (blood/urine) following dietary administration of 5000 ppm pelargonic acid were noted.

Treatment-related morphologic changes were noted in the hearts and livers of rats killed after 28 days. Changes in the heart were also observed in male rats during the recovery phase, but at a lower incidence and severity. Liver lipid content was greater in female rats on diets containing 100, 1000, and 5000 ppm pelargonic acid. The lower body weights in rats killed after 28 days resulted in greater relative weights in a number of organs. Absolute liver weights were greater in male rats that received 5000 ppm pelargonic acid and in female rats that received 1000 ppm and 5000 ppm pelargonic acid in the diet. All other changes that were observed were considered to have been of no toxicological importance. It was concluded that pelargonic acid appeared to have increased the risk of cardiac changes in treated male and female rats and hepatic changes in female rats that received 5000 ppm in the diet. Changes in the liver were not observed at 56 days post-treatment, while cardiac changes persisted at a reduced intensity.

The lowest-observable-effect-level (LOEL) for pelargonic acid was 100 ppm for antemortem data (lower body weights) and 5000 ppm for clinical pathology in rats of both sexes. Taking into consideration the increased liver weights observed after dosing with pelargonic acid, the LOEL was 5000 ppm for male rats and 100 ppm for female rats; the LOEL for macroscopic effects on the liver was 1000 ppm in rats of both sexes. Regarding both cardiac and hepatic effects, the LOEL for macroscopic changes was 1000 ppm (male rats) and 100 ppm (female rats). However, it was noted that because histopathology was not performed on livers from lower dose rats from the scheduled sacrifice, the LOEL for microscopic liver changes may actually be lower than these values.55

A short-term oral toxicity study was conducted using groups of 6 Sprague-Dawley albino rats (3 males, 3 females/group).56 Six groups were fed pelargonic acid in the diet at concentrations ranging from 1500 ppm to 20,000 ppm (one dietary concentration per group) for 2 full weeks. A seventh group (control) was fed untreated feed. Feeding with pelargonic acid did not induce any adverse effects over the range of concentrations evaluated. Body weight gain and food consumption were normal throughout the study. Other than piloerection (not dose-related), no clinical signs were observed. All animals appeared healthy and normal at the time of necropsy. Hematology parameters were all within normal limits, and the same was true for most of the serum clinical chemistry parameters.

Except for the lowest dose group, mean serum alkaline phosphatase activity was significantly greater than the control value. Effects on alkaline phosphatase activity were not considered toxicologically significant relative to liver function, taking into consideration the absence of an effect of pelargonic acid on serum alanine aminotransferase and serum protein content. Total bilirubin was elevated in some of the groups, controls included; high values correlated primarily with the presence of hemolysis in individual blood samples. It was concluded that pelargonic acid did not induce overt signs of toxicity in albino rats, when fed in the diet at concentrations up to 20,000 ppm (2%) for 2 weeks. 56

In another study,36 8 male rats (weights not stated) were fed 4.17% pelargonic acid in the diet (2100 g/kg/day) for 4 weeks. A slight decrease (4%, not statistically significant) in mean growth was observed. No effects on survival were noted.

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Isononyl Isononanoate

The short-term oral toxicity of isononyl isononanoate was evaluated using 4 groups of Sprague-Dawley rats (10 males, 10 females/group) of the Caesarian Obtained, Barrier Sustained-Virus Antibody Free (COBS-VAF®) strain.57 One group served as the vehicle control (corn oil) group, and the 3 test groups received the following doses of isononyl isononanoate (in corn oil) by gavage daily for 4 weeks: 100, 300, and 1,000 mg/kg/day. Test substance- related mortalities were associated with 1 female in the 300 mg/kg/day dose group and 4 females in the 1,000 mg/kg/day dose group. Ptyalism was the only test substance-related clinical sign, and there were no remarkable hematological findings. A correlation between lower body weight gain and lower food consumption was evident only in the highest dose group.

Doses of 300 and 1,000 mg/kg/day were associated with higher enzyme activities, namely aspartate aminotransferase (AST), alanine aminotransferase (ALT), and/or alkaline phosphatase (ALP). Also, compared to controls, the blood urea level was higher in males and females of all dose groups; these changes were attributed to pathological changes in the liver and kidneys that will be mentioned later. Additionally, it was suggested that the high urine volume associated with animals of the highest dose group was related to kidney damage noted at microscopic examination. Higher absolute and relative liver and kidney weights were noted in animals of all dose groups, and lower absolute and relative spleen and thymus weights were observed in 300 and 1,000 mg/kg/day dose groups.

The treatment-related macroscopic findings included enlargement and an accentuated lobular pattern and/or paleness of the liver and a gray/green color of the kidneys in some of the animals from each dose group. Other relevant findings were described as a reduction in size of the spleen and/or thymus in some of the animals from the 300 and 1,000 mg/kg/day dose groups. These findings correlated with contracted spleen and lymphoid depletion in the thymus at microscopic examination, and were considered secondary to the poor physical condition of several treated animals. The treatment-related microscopic findings were described as follows: hepatocellular hypertrophy of the liver (300 and 1,000 mg/kg/day dose groups); liver steatosis (all dose groups); acidophilic globules in cortical tubular epithelium (kidneys), associated with cellular damage in males (all dose groups); vacuolated cortical tubular epithelium in females (300 and 1,000 mg/kg/day dose groups); and contracted spleen and thymic lymphoid depletion (1,000 mg/kg/day dose group).

Isononyl Isononanoate induced mortality at doses of 300 and 1,000 mg/kg/day and liver and kidney (target organs) toxicity in rats at all doses administered. Under the conditions of this experiment, it was not possible to establish a no observed effect level (NOEL) for isononyl isononanoate.57

Comments on the preceding study were received from L’Oreal USA.58 High-fat diets produce adaptive changes in the liver and kidneys of rodents, and, if maintained on high-fat diets for long periods, these changes may develop into a pathologic condition such as fatty liver or steatosis. These authors asserted that steatosis at an isononyl isononanoate dose of 100 mg/kg/day and higher doses in the preceding study should not be considered a toxicological adverse endpoint, but an exacerbation of an adaptive response to administration of a fatty material. Regarding changes in the kidneys, the authors suggested that mineral and other oils are likely to induce acidophilic globules in the kidneys (hyaline droplet nephropathy), due to the sex-linked production of alpha-2-microglobulin, and, understandably, this effect was observed in male rats of all isononyl isononanoate treatment groups. However, because the alpha-2- microglobulin protein (under androgen control) is absent from man and many species, this sex- and species-specific hyaline droplet nephropathy is not considered relevant to man.

Isononyl Alcohol In a study by Rhodes et al.,59 isononyl alcohol was administered by gavage to 5 male rats (Alderly Park Wistar-derived). The test substance (in polyethylene glycol 300) was administered for 14 days at a dose level that was equivalent to 1 mmol/kg/day. Control animals (10 rats) were dosed with polyethylene glycol 300 (10 ml/kg/day). Livers were removed, weighed, and homogenized for enzyme assays. Testis weights were also determined. No major pathological signs of hepatotoxicity resulted from oral dosing with isononyl alcohol. Minor histological changes consisted of slight centrilobular hypertrophy and fat type vacuolation in control and test animals. No effects on body weight gain or testis weight were noted. Isononyl alcohol also did not induce peroxisome proliferation,

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hypocholesteremic/hypotriglyceridemic effects, or effects on catalase. However, compared to controls, isononyl alcohol dosing resulted in slight elevation of palmitoyl CoA oxidase (marker enzyme for peroxisome proliferation).

Neopentyl Glycol A combined repeated dose and reproductive/developmental toxicity study on neopentyl glycol was performed using groups of male and female rats of the Slc: SD strain.28 The test substance, in distilled water, was administered by gavage at doses of 100, 300, or 1,000 mg/kg/day. Control rats were dosed with distilled water. Male rats were dosed over a 42-day period and female rats were dosed from 14 days before mating to day 3 of lactation. There were no dead or abnormal animals with clinical signs related to dosing . Body weight and food consumption data were not indicative of consistent or treatment-related differences between test and control groups. Liver and kidney weights (absolute and relative) were increased in male and female rats of the 300 and 1,000 mg/kg dose groups. Necropsy revealed hypertrophy of the liver in 2 males dosed with 1,000 mg/kg; definite lesions were not found at microscopic examination. A high incidence of protein casts, hyaline droplets, and basophilic change was reported for renal tubules in males dosed with 1,000 mg/kg. The NOAEL for this study was of 100 mg/kg. Results relating to reproductive and developmental toxicity are included later in the report text.

SHORT-TERM DERMAL TOXICITY

Repeated applications of pelargonic acid (25% in mineral oil) to the skin of rabbits in a 28-day study did not cause death, and the random inflammatory changes observed in various organs were described as spontaneous. In other studies, dermal application of pelargonic acid to mice over a 3-day period did not cause death and a TDLo of 3,000 mg/kg was reported. Short-term cutaneous dosing with isononyl isononanoate induced liver and adrenal toxicity in rats at doses up to 860 mg/kg/day.

Pelargonic Acid A 28-day dermal toxicity study60 was conducted using groups of New Zealand White rabbits (5 males, 5 females/group). Weight ranges were 2.1 to 3.2 kg (males) and 2.3 to 3.2 kg (females). Pelargonic acid (25% w/w mixture in mineral oil) was applied to the skin at doses of 500 mg/kg/day (dose volume = 2 ml/kg) daily for a total of 10 applications. The test substance was applied directly to the skin and spread evenly over the test site; patches were not applied. The skin of half of the rabbits per group (3 males, 2 females) was abraded prior to application. The control group was dosed with mineral oil according to the same procedure. For necropsy, 6 rabbits per group (3 with abraded skin and 3 with intact skin) were killed at 2 weeks and surviving animals were killed at 4 weeks.

Slight weight loss (0.1 to 0.4 kg) was noted in most of the rabbits dosed with pelargonic acid after 1 and/or 2 weeks of the study. Weight gain was noted in rabbits that were held for a 2-week recovery period. Slight to moderate weight gains were also noted in vehicle control rabbits. None of the rabbits dosed with pelargonic acid died. Skin reactions are summarized in the section on Skin Irritation later in the report text. Discoloration of the gastric mucosa was observed in treated animals; other gross morphologic findings in treated and/or control animals were not considered treatment-related. Inflammatory changes observed in the kidneys, lungs, and brain and, less frequently, in other organs were not considered treatment-related.60

In a study by Montelius et al.,61 undiluted pelargonic acid (25 µl) was applied to both ears (dorsum) of inbred CBA/Ca mice (groups of 4) once per day for 3 consecutive days. None of the animals died. Murine local lymph node assay (LLNA) results are included in the section on Skin Irritation and Sensitization later in the report text.

Following intermittent dermal application of pelargonic acid to the skin of mice over a 3-day period, the 3 ml/kg dose was the lowest dose that caused a toxic effect (TDLo). In a similar study involving mice, a TDLo of 3000 mg/kg was reported.62

Isononyl Isononanoate

The short-term dermal toxicity of isononyl isononanoate was evaluated using 4 groups of Sprague-Dawley rats (5 males, 5 females/group) of the Caesarian Obtained, Barrier Sustained-Virus Antibody Free (COBS-VAF®) strain 12

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with healthy, intact skin.63 One group served as the vehicle (corn oil) control, and the test groups received cutaneous doses of isononyl isononanoate in corn oil daily for 8 days (860 mg/kg/day dose group) or for 2 weeks (100 and 300 mg/kg/day dose groups). Doses of the test substance or control were applied for 6 h to a 45 to 50 cm2 area on the back (males) or a 30 to 35 cm2 area (females) that had been clipped free of hair. A constant dose volume of 1 ml/kg/day was used. None of the animals died. Slight cutaneous reactions were observed in the 100 and 300 mg/kg/day dose groups (1 animal per group). However, severe skin irritation and necrosis were observed at the application sites of animals of the 860 mg/kg/day dose group and treatment was discontinued after day 8 of dosing.

A correlation between decreased body weight gain and decreased feed consumption was evident only in the highest dose group. Low white blood cell counts were also noted in the highest dose group; however, these changes were considered related to inflammatory reactions and the tissue distribution of inflammatotry cells at the application site. Changes in blood biochemistry were noted in each dose group, all of which were treatment-related. A high urea level and high ALP enzyme activity were noted in 300 and 860 mg/kg/day dose groups. High AST enzyme activity was noted only in the highest dose group, but neither ALP nor AST activity was high in the lowest dose group.

A gray/green coloration of the kidneys was observed in the mid- and high-dose groups, and this finding was correlated with acidophilic globules in the cortical tubular epithelium of high-dose male rats. An accentuated lobular pattern in the liver was noted in all dose groups, and this finding was correlated with steatosis and hepatocellular hypertrophy noted at microscopic examination and considered related to 300 and 860 mg/kg/day doses. Cortical cell hypertrophy in the adrenal glands ranging in severity from minimal to moderate was observed in the highest dose group. Whether or not this finding was considered treatment-related was not stated specifically. The adrenal glands and the liver were considered target organs for isononyl isononanoate toxicity. Under the conditions of this experiment, it was not possible to establish an NOEL for isononyl isononanoate. 63

Comments on the preceding study were received from L’Oreal USA.58 As stated above, study results indicate that steatosis was observed in all isononyl isononanoate dose groups and that acidophilic globules were observed in the kidneys of high-dose male rats. Because isononyl isononanoate is highly lipophilic, the authors considered it highly unlikely that isononyl isononanoate penetrates the skin to any appreciable degree. Furthermore, because some of the effects observed in this dermal study were of a similar order of incidence and severity as those observed at the same dose levels in the short-term oral toxicity study on isononyl isononoate (summarized earlier in report text), these authors suggested that there was significant oral exposure secondary to grooming and licking of the application site.

SUBCHRONIC ORAL TOXICITY

The oral administration of cetearyl isononanoate to rats for 90 days induced reversible fatty alterations in the liver, and an NOAEL of 100 mg/kg/day was reported. The relevance of these findings to man was considered questionable. There were no remarkable gross or microscopic findings in rats fed 1% ethyl pelargonate in the diet for 16 weeks.

Cetearyl Isononanoate A summary of a 1993 subchronic oral toxicity study on Cetiol ® SN (cetearyl isononanoate, % not stated)) was provided by the Personal Care Products Council.64 Three groups of Wistar rats of both sexes received oral doses of 100, 300, and 1000 mg/kg body weight, respectively, over a period of 90 days. A fourth group served as the untreated control. Reversible fatty alterations of the liver were observed in the 1000 mg/kg dose group and in females of the 300 mg/kg dose group. Based on these results, it was determined that the NOAEL should be 100 mg/kg/day. The authors suggested that branched acids, like isononanoic acid, undergo a specific type of metabolism in rodents, and that the fatty alterations in the liver reflect an adaptive response due to increased metabolic activity. The relevance of these changes in the liver to humans was placed in doubt, and, thus, it was anticipated that the NOAEL for human-relevant effects would be > 100 mg/kg body weight.

Ethyl Pelargonate According to Opdyke, 37 no effects were observed at microscopic examination of the following tissues from rats (5 males, 5 females) fed 1% ethyl pelargonate in the diet for 16 weeks: liver, kidney, heart, spleen, testes, viscera, 13

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and hind limb. Terminal hematological examinations and gross pathology and weights of the liver, kidney, heart, spleen, and testes did not differ from findings in control rats.

OCULAR IRRITATION/TOXICITY

The ocular changes observed in a 28-day oral toxicity study (rats) on pelargonic acid were considered sporadic and unrelated to treatment. In studies involving rabbits, pelargonic acid (0.1 ml) was severely irritating or mildly irritating. Neither isononyl isononanoate, ethylhexyl pelargonate (0.1 ml), nor 10% cetearyl isononanoate (0.5 ml) induced ocular irritation in rabbits; however, the cetearyl isononanoate homologue cetyl ethylhexanoate (0.1 ml) and both cetearyl nonanoate and neopentyl glycol diisononanoate (0.1 ml) induced minimal ocular irritation and isononyl alcohol (0.1 ml) induced marked ocular irritation. PEG-5 isononanoate (0.1 ml) induced transient ocular reactions, but was not classified as an ocular irritant.

Pelargonic Acid Results from ophthalmic examinations were presented in the 28-day oral toxicity study on pelargonic Acid in rats,55 summarized in the section on Short-Term Oral Toxicity earlier in the report text. Results of the interim and terminal ophthalmic examinations were as follows: choroidal hypoplasia (1 male - 5000 ppm, 2 females - 100 ppm, 1 female - 10 ppm); retinal hemorrhage (1 female - 100 ppm); vitreous hemorrhage (1 female - 5000 ppm) - all interim results; and hypoplasia (1 male - control); ocular discharge (1 male - 100 ppm, 1 female - control); and choroidal hypoplasia (1 female - 100 ppm) - all terminal results. These findings were considered sporadic and unrelated to treatment.

EPA33 placed pelargonic acid in toxicity category II (corneal involvement or other eye irritation clearing in 8- 21 days), based primarily on results from the following primary ocular irritation study involving 6 Hra: (NZW)SPF adult albino rabbits.65 The undiluted test substance (0.1 ml) was instilled into the right eye of each animal, and untreated left eyes served as controls. Instillation was followed by a 21-day observation period. Pelargonic acid induced corneal and iridial involvement and severe conjunctival irritation. All reactions had cleared by day 21 post- instillation. The average primary irritation score (5-animal mean) was 40.6 at 1 hour and 0 at day 21.

According to STN,62 pelargonic acid was a mild irritant when instilled into the rabbit eye at a dose of 0.1 ml.

Cetearyl Nonanoate The ocular irritation potential of cetearyl nonanoate (97% pure) was evaluated using 3 male New Zealand white rabbits.45 The test substance (0.1 ml) was instilled into the right eye of each animal and reactions were scored at approximately 1 h and 24 h, 48 h, and 72 h post-instillation. Moderate conjunctival irritation was observed; however, there were no changes in the cornea or iris. All eyes appeared normal at 48 h post-instillation. Cetearyl nonanoate was classified as minimally irritating to the rabbit eye.

Cetearyl Isononanoate A summary of a 1970 study evaluating the ocular irritation potential of Cetiol ® SN (cetearyl isononanaote, % not stated) was provided by the Personal Care Products Council. 64 Cetiol ® SN (10% active matter, 0.05 ml) was instilled into the eyes of 2 rabbits, and reactions were scored for up to 72 h post-instillation. The test substance was classified as a non-irritant. In a 1991 study summary provided by the Personal Care Products Council, a homologue, cetyl ethylhexanoate, was applied undiluted (0.1 ml) to the eyes of 3 rabbits and remained for 24 h. Mild conjunctival reactions (erythema, edema, and lacrimation) were observed, all of which had cleared by 72 h post-instillation. Cetyl ethylhexanoate was classified as slightly irritating to the eyes of rabbits.64

Isononyl Isononanoate According to Evonik Industries,47 isononyl isononanoate was not irritating to the eyes of rabbits. Study details were not provided.

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Ethylhexyl Pelargonate The ocular irritation potential of undiluted ethylhexyl pelargonate was evaluated using 6 New Zealand white rabbits.48 Eyes were not rinsed following instillation of the test substance (0.1 ml). Contralateral eyes served as controls. Reactions were scored up to 72 h post-instillation. Ethylhexyl pelargonate was not irritating to the eyes of rabbits.

Neopentyl Glycol Diisononanoate A study evaluating the ocular irritation potential of neopentyl glycol diisononanoate in rabbits was conducted according to the procedure in the preceding section on Cetearyl Nonanoate. 50 Neopentyl glycol diisononanoate (0.1 ml) produced similar results and also was classified as a minimal ocular irritant.

PEG-5 Isononanoate The ocular irritation potential of PEG-5 isononanoate was evaluated using 3 female SPF albino rabbits 51 according to the procedure in the preceding section on Cetearyl Nonanoate. Conjunctival redness and edema had cleared within 14 days, and PEG-5 isononanoate was classified as a non-irritant.

Isononyl Alcohol In a study by Scala and Burtis,39 undiluted isononyl alcohol was instilled (0.1 ml) into the left conjunctival sac of each of 6 rabbits. Untreated eyes served as controls. Ocular irritation reactions were scored using the Draize scale (0 to 110). Draize median irritation scores of 30 (at 24 h) and 2 (at day 7) were reported. The ocular irritation induced by isononyl alcohol was classified as marked. The test substance did not produce severe opacity or other corneal effects, such as sloughing or vascularization.

SKIN IRRITATION

Undiluted pelargonic acid was a mild to severe skin irritant in rabbits and a severe skin irritant in guinea pigs, but was not irritating to the skin of mice. However, undiluted cetearyl isononanoate was non-irritating to the skin of rabbits, but, as undiluted Cetiol ® SN, was slightly irritating to the skin of mice. The remaining studies involved rabbits only. The cetearyl isononanoate homologue, cetyl ethylhexanoate was a slight to moderate skin irritant in rabbits. Isononyl isononanoate and undiluted PEG-5 isononanoate were slightly/mildly irritating and undiluted ethyl pelargonate was moderately irritating to the skin; however, undiluted isononyl alcohol induced marked skin irritation. Skin irritation was not observed following the application of undiluted cetearyl nonanoate or undiluted ethylhexyl pelargonate.

Pelargonic Acid According to Opdyke,37 pelargonic acid (concentration not stated) was classified as a strong skin irritant in guinea pigs, and a moderate irritant when applied undiluted (under occlusion) to abraded or intact skin of rabbits for 24 h.

In a 28-day dermal toxicity study (in Short-term Dermal Toxicity section earlier in report text),60 slight to severe erythema and edema without necrosis or eschar formation were observed in most of the rabbits during the first week of the study. Generally, during the second week, necrosis and eschar formation were observed in all rabbits. Atonia, desquamation, fissuring, and exfoliation were also observed. In rabbits held for recovery, dermal responses subsided. At microscopic examination, epidermal necrosis, hyperplasia, and hyperkeratosis were noted at the application site. Diffuse and perifollicular dermal inflammation were also common. The skin application sites in all surviving animals appeared healed by 2 weeks post-treatment (scheduled recovery period).

EPA33 placed pelargonic acid in toxicity category II (severe irritation at 72 h [severe erythema or edema]), based primarily on results from the following skin irritation study involving 6 adult female albino rabbits of the Hra: (NZW) SPF strain.66 The test substance (undiluted) was applied to intact skin of the back (0.5 ml, exposure area ≈ 6.25 cm2), and the site was covered with a semi-occlusive patch for 4 hours. Reactions were scored up to day 21 post- removal. Skin irritation was observed in all animals; reactions ranged from moderate to severe erythema and edema. 15

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The average of the 4-, 24-, 48-, and 72-hour scores was 5.6 (severely irritating). The following observations were also made at application sites: subcutaneous hemorrhaging, blanching, desquamation, fissuring, possible necrotic areas, denuded areas, and possible scar tissue. With the exception of a denuded area in one animal, all irritation reactions had cleared by day 21.

In a study by Montelius et al.,61 undiluted pelargonic acid (25 µl) was applied to both ears (dorsum) of inbred CBA/Ca mice (groups of 4) once per day for 3 consecutive days. Skin irritation (erythema, edema) was not observed at the test sites. LLNA results are included in the section on Skin Irritation and Sensitization later in the report text.

According to STN,62 pelargonic acid (500 mg dose) was moderately irritating, following application to rabbit skin for 24 h. Undiluted pelargonic acid was classified as a severe irritant, following application to guinea pig skin.

Cetearyl Nonanoate The skin irritation potential of cetearyl nonanoate (97% pure) was evaluated using 3 male New Zealand white rabbits.45 The test substance (0.5 ml) was applied to skin clipped free of hair and the application site was covered with a semi-occlusive patch for 4 h. Reactions were scored 1 h after patch removal and 24 h, 48 h, and 72 h later. Cetearyl nonanoate was classified as a non-irritant.

Cetearyl Isononanoate According to Evonik Industries,46 Cetearyl Isononanoate was not irritating to the skin of rabbits. Study details were not provided. In a 1970 study summary provided by the Personal Care Products Council, repeated applications of undiluted Cetiol ® SN (cetearyl isononanoate, % not stated) were made to the skin of 5 hairless mice for a total of 5 days. It was concluded that the test substance was slightly irritating to the skin.64 In a 1991 study summary provided by the Personal Care Products Council, a homologue, cetyl ethylhexanoate, was applied undiluted (0.5 ml) to dorsal shaved skin of 3 rabbits. The test substance was applied under semi-occlusive conditions for 4 h. Slight to moderate erythema and edema were observed for up to 72 h post-application, and eschar was observed at 1 week post-application. All reactions cleared within 14 days, and cetyl ethylhexanoate was calssified as a slight to moderate skin irritant.64

Isononyl Isononanoate According to Evonik Industries,47 Isononyl Isononanoate was slightly irritating to the skin of rabbits tested according to OECD method 404. Additional study details were not provided.

PEG-5 Isononanoate The skin irritation potential of undiluted PEG-5 Isononanoate was evaluated using 3 male New Zealand white rabbits.51 The test substance (0.5 ml) was applied to skin clipped free of hair and the application site was covered with a semi-occlusive patch for 4 h. Reactions were scored 1 h after patch removal and 24 h, 48 h, and 72 h later. The test substance induced well-defined erythema and very slight edema; no corrosive effects were observed. PEG-5 isononanoate was classified as a mild irritant (PII = 2).

Ethyl Pelargonate According to Opdyke,37 moderate skin irritation was observed after undiluted ethyl pelargonate was applied, under occlusion, to intact or abraded skin of rabbits for 24 h.

Ethylhexyl Pelargonate The skin irritation potential of undiluted ethylhexyl pelargonate was evaluated using 6 New Zealand white rabbits.48 The test substance (0.5 ml) was applied to intact and abraded skin sites that remained occluded for 24 h. Reactions were scored at 24 h and 72 h post-application. Ethylhexyl pelargonate did not induce skin irritation in any of the rabbits (primary irritation index [PII]= 0.40).

Isononyl Alcohol The results of an acute dermal toxicity study39 on undiluted isononyl alcohol (4 rabbits, abraded skin) are summarized earlier in the report text. The test substance was applied under an occlusive binding for 24 h, and the 16

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doses administered ranged from 0.500 to 3.16 g/kg. In this study, dermal irritation (erythema and edema) was classified as marked overall. Both the intensity and duration of skin irritation were dose-related. Atonia and desquamation, with some necrosis or eschar, were persistent findings. It was noted that some of the irritation observed was associated with the trapping of liquid under the occlusive binder at a point where the binder was bound to the animal. It was noted that these areas of intimate contact and pressure gave rise to some of the reported necrosis and eschar.

INFLAMMATION

Ahlfors and Larsson67 conducted a study to examine the tissue response of pelargonic acid in the buccal mucosa of the rat. Both the methyl ester and propyl ester of pelargonic acid (both in acetone) were tested using groups of 6 Sprague-Dawley rats (3 months old). The protocol used consisted of sensitization (dorsal skin) with 2% pelargonic acid (both solutions; dose volume = 100 µl) and challenge in the buccal mucosa (dose volume = 50 µl) with different concentrations of the sensitizing solution (0.2% and 2.0%). The area of the application site (cm2) was not stated. Allergenic potential, as evidenced by the tissue response in the buccal mucosa, was investigated using a skin sensitization procedure and elicitation with 2% or 0.2% solutions. Control rats were exposed to acetone only. The animals were killed 48 h after the last application, and the right buccal mucosa was excised and prepared for microscopic examination. Cellular infiltrates in the buccal mucosa were recorded and compared to normal rat buccal mucosal.

Both test substances (at both concentrations) caused increased cellularity, mainly of the mononuclear cell type. The low concentration of the methyl ester of pelargonic acid (0.2%) induced stronger inflammatory reactions than the high concentration (2.0%). This finding was the opposite of that reported for the propyl ester of pelargonic acid. Both substances were said to have shown a sensitization tendency. Repeated applications of the propyl ester of pelargonic acid (2%) decreased the inflammatory response, when compared to one application. However, for the methyl ester of pelargonic acid, a clear irritative potential was noted with repeated applications. Pre-exposure of dorsal skin prior to buccal painting resulted in an enhanced reaction to pelargonic acid in methyl ester and pelargonic acid in propyl ester.67

COMEDOGENICITY Cetearyl Isononanoate A summary of a 1989 comedogenicity study on Cetiol ® SN (cetearyl isononanoate, % not stated) was provided by the Personal Care Products Council.64 Repeated applications of the test substance to the rabbit ear at concentrations ranging from 10% to 100% did not cause any alterations or produce structures typical of comedogenicity in the infrainfundibulum of hair follicles. The positive control, isopropyl myristate, was comedogenic at a concentration of 10% and non-comedogenic at a concentration of 2%.

SKIN IRRITATION AND SENSITIZATION In a guinea pig RIPT, pelargonic acid (50% in corn oil) was irritating during induction, but was not a sensitizer. Sensitization test results for guinea pigs injected intracutaneously with cetearyl isononanoate (25%) did not differ from those of control guinea pigs. In guinea pig maximization tests, cetearyl nonanoate (10% in sesame oil) and undiluted neopentyl glycol diisononanoate were non-sensitizers; results for 50% cetearyl nonanoate in sesame oil and undiluted neopentyl glycol diisononanoate (for induction) were negative in preliminary skin irritation tests. In mouse LLNAs (for sensitization potential), results were positive at pelargonic acid concentrations of ≥ 50% and ≥ 20% (no- effect-level = 10%) and negative for PEG-5 isononanoate at concentrations up to 100%.

Pelargonic Acid The skin sensitization potential of pelargonic acid was evaluated in a repeated insult patch test using 24 male albino guinea pigs.68 The test group consisted of 10 animals and negative (corn oil) and positive (2,4- dinitrochlorobenzene - DNCB) control groups contained 10 and 4 animals, respectively. During induction, pelargonic acid (50% w/v mixture in corn oil, 0.5 ml) was placed on an adhesive patch (Hill Top Chamber®, 25-mm diameter) that was applied to shaved skin of the anterior left flank for 6 hours per application. Following a non-treatment period, a 6- hour challenge application of the test mixture (0.4 ml) was made to the anterior right flank of each test animal and corn 17

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oil was also applied to a new site on the anterior left flank. The 10 negative control animals were not treated during induction, but received challenge applications of the test mixture and corn oil alone. The positive control was applied during induction and challenge phases.

Pelargonic Acid (50% w/v mixture in corn oil) induced moderate to strong dermal reactions (erythema/edema) in all 10 guinea pigs during the induction phase. Dermal reactions to the mixture were not observed in negative control animals during the challenge phase. The test mixture also did not elicit any dermal reactions in test animals during the challenge phase, and was considered a non-sensitizer in guinea pigs. The positive control induced sensitization.68

In a study by Sikorski et al.,69 3 female BALB/c mice or female CBA/J mice (Harlan) (6 weeks old; weights ≈ 17 to 23 g) were treated with pelargonic acid for 3 consecutive days. The test substance was applied topically (25 µl total/ear; application area [cm2] not stated) to both ears at concentrations ranging from 20% to 80% pelargonic acid in 1-propanol (v/v) and an LLNA was performed according to the procedure of Kimber and Basketter.70 The LLNA measures the proliferative response of lymphocytes in the draining lymph nodes during the induction of contact sensitization by epicutaneous exposure to an allergen.

Compared to the vehicle control, pelargonic acid produced slight increases in the % of B220+ lymphocytes at all doses. These findings were not dose-related. Even at high concentrations, the cell number per node and the % of B220+ cells never approached the values that were associated with allergens such as 1-chloro-2,4,6-trinitrobenzene (TNCB) and 1-chloro-2,4-dinitrobenzene (DNCB).69

In an effort to develop a predictive in vitro assay for the identification of potential contact allergens, Pistoor et al.71 compared the effects of epicutaneous application of known contact allergens, pelargonic acid and other known irritants, and dermatologically inactive chemicals on Langerhans cells in skin biopsy cultures from 7 healthy donors. The skin specimens were obtained from breast reduction surgery or abdomen reduction surgery. Pelargonic acid (80% v/v) in propane-1-ol and 100% propylene glycol was tested. Untreated and treated skin samples were cultured for 24 h. As controls, explants were topically treated with the solvent.

Immunohistochemical analysis of cryostat sections of all biopsies treated with contact allergens showed the following: (1) a large decrease in the number of Langerhans cells in the epidermis, as evaluated by a decrease in human leukocyte antigen (HLA)-DR+- expressing cells and CD1a+- expressing cells. (2) accumulation of the remaining Langerhans cells at the epidermal-dermal junction. Pelargonic acid did not induce these changes.71

Montelius et al.61 evaluated pelargonic acid, neat or in dimethylformamide, in the LLNA. Pelargonic acid showed a dose-response relationship and, clearly, positive results when tested at concentrations of ≥50%. Thus, this chemical was classified as a potential sensitizer. Short-term oral toxicity results for inbred CBA/Ca strain mice used in this assay are summarized under that subheading earlier in the report text. Similarly, results relating to skin irritation are summarized in the Skin Irritation section.

Woolhiser et al.72 combined the parameters of an LLNA and a mouse ear swelling irritancy assay in an effort to establish a single, rapid screening procedure for the sensitization and irritation potential of chemicals. Groups of 5 female B6C3F1 mice (C57BL/6 x C3HHeN; weights = 18 to 26 g) were used. Pelargonic acid was administered to the dorsal and ventral surfaces of each ear at concentrations ranging from 5% to 60% (in acetone) for 4 consecutive days. Compared to the vehicle control, pelargonic acid produced a dose-dependent and statistically significant increase in lymph node cell proliferation at concentrations of 20%, 40%, and 60%. The no-effect-level was at a concentration of 10%. Known sensitizers (i.e., oxazolone; 2,4-dinitrofluorobenzene; and toluene diisocyanate) evaluated in the assay produced marked lymph node cell proliferation.

Cetearyl Nonanoate The skin sensitization potential of cetearyl nonanoate (89% pure) in sesame oil was evaluated in a maximization test using 15 male guinea pigs.45 Of the 15, 5 comprised the vehicle control group. Because topical induction with 50% cetearyl nonanoate in sesame oil did not induce skin irritation in a preliminary experiment, the skin was treated with sodium lauryl sulfate (SLS) in order to induce local irritation. Cetearyl nonanoate (10% in sesame oil) was administered during the first (intracutaneous) induction stage, and reactions were evaluated after 24 and 48 h. 18

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During the second (topical) induction stage, cetearyl nonanoate (50% in sesame oil, 2 ml) was applied and reactions were scored 48 and 72 h after the initiation of exposure. At day 21, the animals were challenged with 10% cetearyl nonanoate in sesame oil (2 ml). Skin changes were not observed following intracutaneous induction or during the challenge phase, and the same was true for the vehicle control. Cetearyl nonanoate was classified as a non-sensitizer.

Cetearyl Isononanoate A summary of a 1971 non-GLP-sensitization study on Cetiol ® SN (cetearyl isononanoate, % not stated) was provided by the Personal Care Products Council.64 During induction and challenge phases, the test substance (25%) was injected intacutaneously (10 injections within 14 days) into 5 male guinea pigs of the Pirbright White strain. The skin reactions observed in test animals did not differ from those observed in the control group.

Neopentyl Glycol Diisononanoate A maximization test on neopentyl glycol diisononanoate was performed according to a slight modification of the preceding test procedure.50 Undiluted test material was applied during the second induction and challenge phase. Initially, the skin was treated with SLS because topical induction with undiluted neopentyl glycol diisononanoate did not induce skin irritation in a preliminary experiment. Neopentyl glycol diisononanoate was classified as a nonsensitizer.

PEG-5 Isononanoate The skin sensitization potential of PEG-5 isononanoate in CBA/Ca mice (groups of 4) following topical application was evaluated in the local lymph node assay.51 The undiluted test substance and concentrations of 25% and 50% in acetone/olive oil were applied to the dorsal surface of the ear. The control group was treated with vehicle only. The stimulation index (SI) was expressed as the mean radioactive incorporation for each treatment group divided by the mean radioactive incorporation of the vehicle control group. SI values of 1.70 (25% concentration), 2.42 (50%), and 1.85 (100%) were reported, and PEG-5 isononanoate was classified as a non-sensitizer.

REPRODUCTIVE AND DEVELOPMENTAL TOXICITY

Daily doses of pelargonic acid up to 1,500 mg/kg/day did not induce reproductive effects in inseminated female rats. Results from other studies support pelargonic acid daily doses of 1500 mg/kg/day as the no-observable- effect-level (NOEL) for maternal/developmental toxicity in rats, and isononyl isononanoate daily doses of 300 mg/kg/day as the NOEL for developmental toxicity in rats. In a teratogenicity study on cetearyl isononanoate, the NOAEL for maternal toxicity and embryo-/fetotoxicity in rats was 1000 mg/kg body weight. Two branched-chain nonanols (perhaps incorrectly identified as isononanol) caused a marked degree of maternal and fetal toxicity in rats at daily doses of 7.5 and 10 mmol/kg/day (1,185 and 1,580 mg/kg/day) and slight fetal effects at 5 mmol/kg/day doses. Neopentyl glycol at oral doses up to 1,000 mg/kg/day did not induce reproductive effects.

Pelargonic Acid A study by Hazleton Labs America Inc.36 conducted to determine the appropriate dose level of pelargonic acid for a teratology screening study involving Cesarean-derived, Sprague-Dawley rats. Details relating to the conduct of this study are included in the section on Short-Term Oral Toxicity earlier in the report text. The number of corpora lutea per ovary and the number and placement of uterine implantations, resorptions, and live and dead fetuses were recorded. Mean ovarian and uterine weight data were comparable between treated and control groups. No treatment- related reproductive effects were noted over the range of administered doses (1.0 to 10.0 ml/kg).

A study designed to evaluate the embryo/fetal toxicity and teratogenic potential of pelargonic acid was conducted using groups of 22 mated female Crl:COBS, CD®(SD)BR rats (14 weeks old).73 Females of the test group were dosed orally (by gavage) with pelargonic acid (in corn oil; dose = 1500 mg/kg) on gestation days 6 through 15. The control group received corn oil according to the same procedure. Pregnant females were killed on day 20 and fetuses were delivered by cesarean section. Neither test substance-related maternal toxicity nor effects on food and water consumption were observed in the test or control group. Additionally, there was no definitive evidence of teratogenic effects in the test or control group. 19

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EPA33 reported the results of a developmental toxicity study involving rats. Treatment of the animals with pelargonic acid had no adverse effects on clinical signs, body weight gain, or food/water consumption. Fetal toxicity was not observed in treated rats or in untreated control rats, and the following parameters were comparable between treated and control rats: mean number of viable fetuses, early or late resorptions, implantation sites, corpora lutea, pre- and post-implantation losses, sex ratios, and fetal body weights. The no-observed-effect level (NOEL) for maternal and developmental toxicity was 1500 mg/kg/day, and the LOEL was greater than 1500 mg/kg/day.

Cetearyl Isononanoate A summary of a 1993 teratogenicity study on Cetiol ® SN (cetearyl isononanoate, % not stated) was provided by the Personal Care Products Council.64 Three groups of pregnant CD rats received oral doses (gavage; dose volume = 10 ml/kg) of 100, 300, and 1000 mg/kg body weight, respectively, from day 6 to day 15 of gestation. A fourth group served as the untreated control. None of the animals died and maternal body weight gain was not affected by treatment. All of the females had viable fetuses, and pre-implantation loss and mean numbers of resorptions were not affected by treatment. Non-dose-related post-implantation loss was observed in treatment groups. All parameters were said to have been comparable to those of the control group. The results of skeletal and visceral examinations did not provide evidence of any treatment-related malformations. The NOAEL for maternal toxicity and embryo-/fetotoxicity was 1000 mg/kg body weight.

Isononyl Isononanoate

The developmental toxicity of isononyl isononanoate (in corn oil) was evaluated using groups of 10 mated female Sprague-Dawley rats.74 The 3 test groups received doses of 30, 100, and 300 mg/kg/day, respectively, by gavage on day 6 to day 17 post-coitum. The control group was dosed with corn oil. There was no evidence of treatment-related, macroscopic post-mortem findings in any of the females, and none of the animals died. There was also no evidence of total resorption or abortion. The number of implantation sites and corpora lutea per female was similar in all dose groups, and, compared to controls, the number of resorptions (early and late) and post-implantation loss per female in either dose group were similar. Additionally, there were no differences in the number of live fetuses in either dose group when compared to controls, and neither external anomalies nor malformations were observed. It was concluded that isononyl isononanoate did not induce direct embryotoxicity or fetotoxicity at doses up to 300 mg/kg/day.

Isononyl Alcohol Hellwig and Jäckh75 studied the developmental toxicity of isononyl alcohol using sexually mature, virgin Wistar rats of outbred strain Chbb/THOM. The 2 types of isononyl alcohol, both identified as CAS No. 68515-81-1, tested were defined as follows: Isononanol type 1 (purity ≥ 99%) – of commercial origin and consisted of roughly equivalent amounts of 3,4-, 4,6-, 3,6-, 3,5-, 4,5-, 5,6-dimethylheptanol-1. Isononanol type 2 (purity ≥ 99%) – produced at BASF and the following were listed as main components: 4,5-diemthylheptanol-1 (~23%), 4-methyloctanol-1 (29%), 3-ethylheptanol-1 (3%), 6-methyloctanol-1 (15%), and 3-ethyl-4-methylhexanol (1%) [Note: Based on these chemical composition data, there is reason to believe that neither chemical (type 1 or 2) is isononanol. However, it is evident that both are branched-chain nonanols]. The test substances were diluted (twice-distilled water, employing ~ 0.005% Cremophor EL [PEG-35 Castor Oil] as emulsifier) to a standard dose volume of 5 ml/kg body weight. Each test substance was administered by gavage (doses ranging from 1 to 10 mmol/kg/day) to pregnant females (10/group) on days 6 to 15 post-coitum. Two control groups were treated with either doubly-distilled water alone (control group 1) or water plus ~ 0.005% Cremophor EL (control group 2).

Both isononanols (types 1 and 2) exhibited a marked degree of maternal and fetal toxicity at daily doses of 7.5 and 10 mmol/kg/day, and slight fetal effects at 5 mmol/kg/day doses. Of the fetal findings (malformations, variations, or retardations), the only ones that were significantly different from controls were the number of fetuses with skeletal retardations in the 5 mmol/kg/day dose group (p < 0.1, both control groups) and the number with skeletal variations in this group (p < 0.05; 1 control group). Dosing at 1 mmol/kg/day did not cause adverse effects. When pregnant females were dosed with 7.5 mmol/kg /day (isononanol type 1) in a supplementary experiment, the incidence of malformations (mainly related to the heart) was statistically significantly increased (p < 0.01; 1 control group). Resorptions and post- 20

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implantation loss were also significantly increased (p < 0.01; 1 control group) at this dose level. For isononanol type 2, the only significant fetal findings (7.5 mmol/kg/day doses) were the number of fetuses with skeletal retardations (p < 0.01, 0.05; 1 control group – both values), number of fetuses with skeletal variations (p < 0.051 1 control group), number of fetuses with variations (p < 0.05; 1 control group) and number of fetuses with malformations (p < 0.05; 1 control group). Resorptions were also significantly increased (p < 0.05; both control groups).

Neopentyl Glycol A combined repeated dose and reproductive/developmental toxicity study on neopentyl glycol was performed using groups of male and female rats of the Slc: SD strain.28 The test substance, in distilled water, was administered by gavage at doses of 100, 300, or 1,000 mg/kg/day. Control rats were dosed with distilled water. Male rats were dosed over a 42-day period and female rats were dosed from 14 days before mating to day 3 of lactation. There were no test substance-related effects on copulation, fertility, or the estrous cycle of rats, and the same was true during the lactation period. With the exception of one control rat, delivery was normal for all dams. There were no test substance-related abnormal findings in any of the pups delivered. The body weight gain of pups was normal up to day 4 of lactation. Test substance-related, abnormal gross findings were not reported for stillborn, dead pups or pups killed at day 4 of lactation. Additionally, no developmental toxic effects were associated with test substance administration. The NOAEL for neopentyl glycol (P and F1 generations) was 1,000 mg/kg. Results relating to the short-term oral toxicity of neopentyl glycol are included earlier in the report text.

GENOTOXICITY

While one mammalian cell assay of pelargonic acid was positive with metabolic activation (doses up to 600 µg/ml), it was negative without metabolic activation. All other bacterial, mammalian cell, and in vivo assays for pelargonic acid (doses up to 5,000 µg/plate or 5,000 mg/kg) and the following pelargonic acid esters (doses up to 5,000 µg/plate) were negative: cetearyl isononanoate, cetearyl nonanoate, ethylhexyl isononanoate, isononyl isononanoate, neopentyl glycol diisononanoate, and PEG-5 isononanoate. Negative results were also reported for neopentyl glycol at doses up to 5,000 µg/plate (bacterial cells)and up to 5 mg/ml (mammalian cells).

Pelargonic Acid In an Ames test,76 the mutagenicity of pelargonic acid was evaluated with and without metabolic activation using Salmonella typhimurium strains TA98, TA100, TA1535, TA1537, and TA1538. The test substance (in dimethyl sulfoxide [DMSO]) was evaluated at doses ranging from 100 to 5,000 µg/plate in this reverse mutation assay. Pelargonic acid did not cause a positive increase in the number of histidine revertants per plate in any of the tester strains, with or without metabolic activation, and, therefore, was nonmutagenic.

The mutagenicity of pelargonic acid was evaluated in a forward mutation assay, using the L5178Y mouse lymphoma cell line.77 In preliminary cytotoxicity assays, pelargonic acid (in DMSO) induced dose-related cytotoxicity, and was completely toxic at a concentration of 4,000 µg/ml (without metabolic activation) and at a concentration of 2,000 µg/ml (with metabolic activation). In forward mutation assays without metabolic activation (concentrations of 150 to 1600 µg/ml), pelargonic acid did not induce repeatable increases in the mutant frequency. In assays with metabolic activation, dose-related increases in the mutation frequency that exceeded the minimum criterion for a positive response were observed at concentrations ranging from 50 to 600 µg/ml. These increases were primarily due to increases in small colonies, considered to reflect gross chromosomal changes rather than small changes within a gene. Results for pelargonic acid were positive with metabolic activation, but negative without metabolic activation.

The mutagenicity of pelargonic acid was evaluated in the in vivo micronucleus assay.78 Three groups of ICR mice (5 males, 5 females/group) received pelargonic acid, in corn oil, at oral doses of 1250, 2500, and 5,000 mg/kg, respectively. After dosing, the animals were killed and bone marrow was extracted. Pelargonic acid did not induce a significant increase in micronuclei in bone marrow polychromatic erythrocytes, and was considered non-mutagenic in this assay.

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The National Toxicology Program79 evaluated the mutagenicity of pelargonic acid (in dimethylsulfoxide) in Salmonella typhimurium strains TA 98 and TA 100 using the Ames preincubation assay. Pelargonic acid was tested at doses up to 5000 µg/plate with and without metabolic activation, and results were negative in each strain.

Cetearyl Nonanoate The mutagenicity of cetearyl nonanoate (in acetone; doses up to 5,000 µg/plate) was evaluated in the Ames test using Salmonella typhimurium strains TA1535, TA1537, TA102, TA98, and TA100.45 Results were negative with and without metabolic activation.

Cetearyl Isononanoate A summary of a 1991 genotoxicity study on Cetiol ® SN (cetearyl isononanoate, % not stated) was provided by the Personal Care Products Council.64 The test substance was evaluated at concentrations up to 5000 µg/plate with and without metabolic activation. Neither toxicity nor reverse mutations were observed over the range of concentrations tested, and Cetiol ® SN was classified as nonmutagenic.

Ethylhexyl Isononanoate

The mutagencity of ethylhexyl isononanoate (in acetone; doses up to 5,000 µg/plate) was evaluated in the Ames test using the bacterial strains stated immediately above.80 Results were negative with and without metabolic activation.

Isononyl Isononanoate

The mutagenicity of isononyl isononanoate (in ethanol) was evaluated using the following Salmonella typhimurium strains with and without metabolic activation: TA1535, TA1537, TA98, TA100, and TA102.81 Anthramine served as the positive control for assays with metabolic activation and the following chemicals served as positive controls for non-activation assays: sodium azide, 9-aminoacridine, 2-nitrofluorene, and mitomycin C. Isononyl isononante was not mutagenic at doses up to 5,000 µg/plate either with or without metabolic activation. All positive controls were mutagenic.

Neopentyl Glycol Diisononanoate The Ames test was also used to evaluate the mutagenicity of neopentyl glycol diisononanoate (in acetone; doses up to 5,000 µg/plate) in the Salmonella typhimurium strains mentioned in the preceding study.50 Results were negative with and without metabolic activation.

PEG-5 Isononanoate A battery of mutagenicity tests on PEG-5 isononanoate was performed.51 Ames test results (doses up to 5,000 µg/plate ± metabolic activation) were negative in the Salmonella typhimurium strains mentioned in the preceding section on Cetearyl Nonanoate. The mutagenicity of a formulation containing water, trideceth-9, and 29% PEG-5 isononanoate was evaluated in a chromosomal aberration assay involving human lymphocytes in vitro. The highest test concentration of the formulation was 5,000 µg formulation /ml (1450 µl PEG -5 isononanoate /ml). Dose-dependent increases in chromosomal aberrations, with metabolic activation, were within the range of the laboratory’s historical control data, and, thus, considered biologically irrelevant. Clastogenicity was not observed, either with or without metabolic activation. The mutagenicity of the same formulation and maximum test concentration was evaluated in the mammalian cell gene mutation test (mouse lymphoma assay) using the L5178Y/TK+/- cell line with and without metabolic activation. Results were negative with and without metabolic activation.

Neopentyl Glycol The mutagenicity of neopentyl glycol was evaluated using Salmonella typhimurium strains TA100, TA 1535, TA98, and TA1537 and Escherichia coli strain WP2 uvrA.28 Mutagenicity was evaluated at doses up to 5,000 µg/plate with and without metabolic activation. The minimum dose at which toxicity to bacteria was observed, with and without metabolic activation, was > 5,000 µg/plate. Results for neopentyl glycol were classified as negative in this assay. The 22

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mutagenicity of neopentyl glycol was also evaluated in an assay involving Chinese hamster CHL cells. Test substance (in distilled water) doses up to 1.0 mg/ml were evaluated, and results were classified as negative.

Kusakabe et al.82 provided Ames test results and the results of a chromosomal aberration test, using Chinese hamster lung (CHL/IU) cells, on neopentyl glycol. In the latter assay, proliferating cells were treated with neopentyl glycol for 6h (short-term) with and without metabolic activation. These cells were also treated with neopentyl glycol for 24 h and 48 h continuously without metabolic activation. Ames test results were negative. In the chromosomal aberrations test, results for neopentyl glycol were negative at doses manifesting 50% or < 50% cytotoxicity (or at 5 mg/ml or 10 mM). Negative chromosomal aberrations test results (with and without metabolic activation) were associated with short-term as well as continuous treatment assays.

Methyl Pelargonate Renner83 evaluated the anticlastogenic potential of methyl esters of fatty acids in vivo in the chromosomal aberration assay using Chinese hamster bone marrow cells. Chinese hamsters of both sexes were gavaged (single oral dose) with the methyl ester of pelargonic acid, followed immediately by dosing with the mutagenic alkylating agent, bisulfan (1,4- butandiolbis (methane sulphonate). The chromosome-breaking activity of bisulfan was not modulated by the methyl ester of pelargonic acid (C9) and other short-chain fatty acids. However, the methyl esters of fatty acids ranging from lauric acid (C12) up to nonadecanoic acid (C19) reduced the rate of aberrant metaphases from 9.4 % to ∼3% at doses of 100 mg/kg and less.

CARCINOGENICITY

The dermal carcinogenicity of undiluted pelargonic acid was evaluated using groups of 50 male C3H/HeJ mice.84 Pelargonic acid (dose = 50 mg/kg; was applied twice weekly to interscapular skin (clipped free of hair) of the back for 80 weeks or until a neoplasm was grossly diagnosed as an advanced tumor. All surviving mice were killed between 80 and 83 weeks. Other groups included in the study were an untreated control group, a group treated with mineral oil, and a positive control group (0.05% benzo(a)pyrene in mineral oil). Sixty-six percent of the mice in the Pelargonic Acid-treated group survived to week 78, compared to 52% and 64% for untreated and mineral oil controls, respectively. None of the positive control mice receiving applications of 0.05% benzo(a)pyrene survived to week 78. Forty-two mice treated with pelargonic acid lived long enough to have sufficient exposure to develop a tumor within the average latent period.

The following non-neoplastic skin lesions were observed in pelargonic acid-treated mice: ulcer (7 mice), skin pigmentation (41 mice), fibrosis (48 mice), scar formation (14 mice), acanthosis (48 mice), and hyperkeratosis (40 mice). The authors noted that hyperplasia of the dermis (fibrosis), acanthosis, and hyperkeratosis are common findings in areas of mouse skin that have been clipped free of hair. There was no evidence of gross skin tumors in pelargonic acid-treated mice or in the 2 control groups. Gross skin tumors were reported for 46 positive control mice. The incidence of hepatocarcinomas in the test group was at least as high as that in the negative control groups after 80 weeks.84

ANTITUMOR ACTIVITY

Pelargonic Acid Nishikawa et al.85 studied the antitumor activity of pelargonic acid using groups of 6 female ddY mice (weights = 23 ± 2 g) inoculated i.p. with Ehrlich ascites carcinoma. Pelargonic acid (in saline with 0.2% Tween 80) was injected i.p. once daily for 5 consecutive days. Six control mice were injected with the vehicle according to the same procedure. None of the animals died. Pelargonic acid did not induce antitumor activity.

Ethyl Pelargonate According to Opdyke,37 ethyl pelargonate (1.0 mmol/ml) induced antileukemic activity when an in vitro mixture with AKR leukemic cells was injected subcutaneously (s.c.) into male AKR mice.

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CLINICAL ASSESSMENT OF SAFETY

SKIN IRRITATION

Pelargonic acid is a known skin irritant, based on the results of both predictive and provocative human skin irritation studies. In predictive tests, pelargonic acid induced skin irritation at concentrations ranging from 5% to 80%; ethyl pelargonate was a skin irritant at a concentration of 20%, but not 12%. Predictive skin irritation test results for undiluted cetearyl nonanoate, 20% cetearyl isononanoate, and undiluted neopentyl glycol diisononanoate were negative.

Predictive Tests

Pelargonic Acid Stillman al.86 conducted a cumulative irritation study involving 20 male subjects, where the relative irritancy of free fatty acids of different chain lengths was evaluated. Equimolar concentrations (0.5 M and 1.0 M) of even- and odd-numbered - straight chain saturated fatty acids were dissolved in propanol. Each Al-test® patch containing a fatty acid (0.5 M) was applied to the interscapular area of 10 subjects, and, similarly, each fatty acid was applied at a higher concentration (1.0 M) to the remaining 10 subjects. A control patch containing propanol was also applied to each subject. Patches remained in place for 24 h and reactions were scored 30 minutes after patch removal. This procedure was repeated daily for a total of 10 applications. In another test, each fatty acid (0.01 M and 0.1 M, in olive oil; injection volume = 0.1 ml ) was injected into the backs of 5 additional subjects. Olive oil (control) was injected intradermally into each subject. Test sites were evaluated for erythema and induration at 24h and 48 h.

In most men in the dermal study, an erythematous response to the 0.5 M fatty acid concentration was observed by day 10 for fatty acids in the C8 through C12 range. Seven of 10 subjects had an erythematous reaction to pelargonic acid (0.5 M). For the 1.0 M fatty acid concentration, an erythematous response was observed in all 10 subjects by the eighth day of application of fatty acids in the C8 through C12 range. In both groups of 10 subjects, there were no reactions to propanol. Following intradermal injection, slight induration (5 mm) and mild erythema were observed at all injection sites in each of the 5 subjects (0.01 M fatty acids, 0.1 M fatty acids, and olive oil control). It was not possible to differentiate the control sites from the test sites.86

In a study by Wahlberg and Maibach,87 116 healthy male subjects (21 to 55 years old) were patch tested with pelargonic acid at concentrations of 5%, 10%, 20%, and 39.9% in propanol and a propanol-treated control patch was used. Dose response curves were developed. Patches (Al-test® discs) were saturated with 0.04 ml of a test solution and applied to the upper back for 48 h. Reactions were scored at 48 h and 96 h post-application. Pelargonic acid at concentrations of 20% and 39.9% in propanol caused skin irritation in > 90% of the subjects tested at both 48 h and 96 h post-application. The lesions consisted primarily of erythema at 48 h and pigmentation at 96 h. Pelargonic acid (10 % in propanol) caused skin irritation in 54.3% of the subjects at 48 h post-application and in 48.5% of the subjects at 96 h. Pelargonic acid (5% in propanol) caused skin irritation in 12.9% of the subjects at 48 h post-application and in 13.9% of the subjects at 96 h post-application.

In a study by Agner and Serup,88 16 volunteers (10 females, 6 males; median age of 29.5 years) were patch tested (closed patches, Finn chambers) with 20% pelargonic acid in propanol (pH of 4.3). Propanol was one of the controls used. Patches were applied to the anterolateral surface of both upper arms for 24 h. Reactions were scored at 24, 48, and 96 h post-application according to the following scale: 0 (no reaction) to 3 (strong positive reaction: marked erythema, infiltration, possibly vesicles, bullae, pustules and/or pronounced crusting). Pelargonic acid caused erythema and slight infiltration at 24 h post-application and, occasionally, slight crusting was observed at 48 h. Scores ranging from 0 to 3 were reported at 24 h, and scores ranging from 0 to 2 were reported at 48 h and 96 h.

Willis et al.89 conducted a skin irritation study using 42 healthy, non-atopic male volunteers (mean age = 34 years; skin types: II [20 subjects], III [17 subjects], and IV [5 subjects]). Pelargonic acid was patch-tested (Finn chambers, volar forearm) at the following concentrations (in propan-1-ol): 40% (12 subjects), 60% (32 subjects), 70% (32 subjects), and 80% (28 subjects). Each subject received between 3 and 10 patch tests. The patches remained in 24

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place for 48 h, and reactions were scored 1 h later according to the following scale: - (no visible reaction) to 4+ (intense erythema with bullous formation). Propanol was one of the controls used. The following positive reactions were reported: 2 of 12 subjects (40% pelargonic acid; faint, patchy erythema [+/-]), 20 of 32 subjects (60% pelargonic acid; mostly erythema with edema [1+]), 22 of 32 subjects (70% pelargonic acid; mostly erythema with edema [1+]), and 28 of 28 subjects (80% pelargonic acid; mostly erythema with edema [1+] and erythema with edema and papules [2+]).

Agner and Serup90 patch tested 16 healthy subjects (ages not stated) with pelargonic acid (20% in propanol). Closed patches (Finn chambers) containing the test substance were applied to the anterolateral surface of both upper arms. The patches were removed at 24 h post-application and reactions were scored at 24 h and 96 h post-application. Propanol (solvent) was also evaluated in this study. A mean irritation score of 2 (moderate positive reaction) was reported at 24 h and a mean score of 1 (weak positive reaction) was reported at 96 h.

In a study by Willis et al.,91 10 healthy, nonatopic male volunteers (mean age = 35) were patch tested (Finn chambers) with 80% (w/v) pelargonic acid in propan-1-ol (w/w) 100% propylene glycol and the vehicle control. The test substance was applied to the volar aspect of the forearm for 48 h, and reactions were scored at 1 h post-removal according to the following scale: negative to severe (erythema, edema, and vesiculation). Punch biopsy specimens were obtained from the application site and an untreated site and examined microscopically. Patch test results for the vehicle control were negative. Pelargonic acid induced mild to moderate skin irritation. At microscopic examination, the most notable feature in all skin samples was the presence of “tongues” of dense, irregularly shaped eosinophilic keratinocytes containing shrunken nuclei that extended down from the stratum granulosum and into the stratum spinosum. The cytoplasm of these cells was composed largely of dense, wavy aggregates of osmiophilic keratin filaments that were associated with prominent intercellular desmosomes.

Agner and Serup92 investigated pathophysiological components of irritant contact dermatitis induced by 20% pelargonic acid in propanol using 20 healthy volunteers (12 males, 8 females; mean age = 27.5 years). A closed patch (Finn chamber) containing the test material was applied to the flexor side of both upper arms for 24 h. Reactions were scored at 24 h (30 to 60 min after patch removal), 48 h, and 96 h post-application. At 24 h, skin irritation (scores not included) was observed in all 20 subjects. At 48 h, severe reactions to pelargonic acid were not observed. A significant increase in transepidermal water loss (p < 0.01) was observed after 24 h, 48 h, and 96 h, when compared to propanol (control) patch test areas. However, the increase was not significant after 24 h when compared to empty chamber areas. Blood flow was significantly increased (p < 0.01) in areas patch-tested with pelargonic acid after 24 h, 48 h, and 96 h post-application, when compared to propanol and empty chamber controls. Skin thickness was also significantly increased (p < 0.01) throughout the observation period when compared to controls.

In another study, Agner and Serup93 investigated a possible seasonal variation in the skin response to pelargonic acid during the winter and summer. The study was conducted using 17 healthy volunteers (10 males, 7 females; mean age = 27 years). The test substance (20% pelargonic acid in propanol) was applied (closed patch, Finn chamber) to each arm for 24 h. Reactions were scored at 30 min post-removal. Two subjects had reactions that ranged from a score of 0.5 to 2 during the winter and 1 subject had reactions that ranged from 1 to 2 during the summer. Reactions were not observed at sites treated with propanol, water, or to which an empty chamber was applied. For sites tested with pelargonic acid and control patch sites, the hydration state of the superficial epidermis was significantly lower (p < 0.01) during the winter period, compared to the summer period. No significant seasonal variation was found in the skin response (irritation or transepidermal water loss) to pelargonic acid.

Willis et al.94 patch tested 10 healthy, non-atopic male volunteers with 80% (v/v) pelargonic acid (in propan-1- ol, 100% propylene glycol). Ages of the test subjects were not stated. Patches (Finn chambers) were applied to the volar aspect of the forearm and removed at 48 h. Reactions were scored at 1 h post-removal according to the following scale: 0 (negative) to 3 (erythema, edema, and vesiculation). Punch biopsies were obtained. Most of the irritation reactions were classified as mild to moderate. A statistically significant decrease (p < 0.01) in the epidermal density of CD1+ cells and the length of dendritic processes of these cells (p < 0.001) was noted. There was also morphologic evidence of cellular injury to Langerhans cells in the majority of biopsies.

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Reiche et al.95 patch tested 152 female subjects (> 18 years old; 37 were atopic) with pelargonic acid in propan-1-ol at concentrations of 10%, 20%, 40%, and 80%. The test substance (each concentration; 15 µl aliquot) was soaked onto a filter paper disc in each Finn chamber (8 mm diameter) - applied to the right and left lower back. The patches were removed at 47 h and reactions were scored at 48 h and 96 h. Propan-1-ol was applied to control sites. Erythema was more severe at 48 h than at 96 h (p < 0.001). With the exception of 20% pelargonic acid, decreased erythema with time was noted at each concentration. Changes at the skin surface (e.g., dryness, wrinkling, or necrosis), however, increased with time at the higher concentrations. For example, at a concentration of 80% pelargonic acid, surface changes were significantly greater at 96 h than at 48 h (p < 0.0001). The same was true for 40% pelargonic acid (p = 0.0015).

In a study by Clemmensen et al.,96 the forearm skin of healthy volunteers was exposed in 2 different cumulative skin irritation test models, a repeated open model (ROAT) and an exaggerated wash test model. In the ROAT, 12 healthy volunteers (7 males, 5 females; mean age = 25.3 years) were tested with pelargonic acid at concentrations of 10%, 20%, and 30% (vehicle = propanol). Sodium lauryl sulfate (SLS) was tested at concentrations of 0.5%, 1.0%, and 2.0% (vehicle = sterile water). Applications were made to volar forearm skin of the right arm (SLS) and left arm (pelargonic acid). Both substances were applied (10-min application) daily for 5 + 4 days (no applications on weekend).

The wash test (24 healthy volunteers) involved the induction of irritation by 3 daily washings for 6 days and the maintenance of the dermatitis by 2 daily washing for 12 days with pelargonic acid concentrations of 30%, 40%, and 50% (vehicle = propanol) or SLS concentrations of 5%, 10%, and 15% (vehicle = sterile water). Washing with pelargonic acid involved 8 female and 4 male subjects (mean age = 27 years). Test areas were on the volar aspect of each forearm and an oral hygiene sponge was used for washing. Erythema/edema were graded according to a total scoring range of 0 to 15 (scores > 9 = very strong or caustic reaction).

In the ROAT, the maximum observed visual score of 12 was reported for 2 subjects tested with 30% pelargonic acid. At the concentrations tested, pelargonic acid induced more irritation when compared to SLS and was considered quite irritating. Dose-dependent healing and irritation scores approaching the pre-irritation values were reported during follow-up at day 17. In the wash test model, all subjects treated with pelargonic acid achieved the desired irritation score of 2 for at least one concentration of pelargonic acid. The maximum observed irritation score of 12 (1 subject,15% SLS) was observed at day 9.96

Suihko and Serup97 studied the applicability of fluorescence confocal laser scanning microscopy for in situ imaging of irritant contact dermatitis caused by pelargonic acid using 12 healthy individuals (8 males, 4 males; 18 to 64 years old). Using Finn chambers (occlusive patches), the flexor side of the right and left forearm was exposed to 60 µl of 10% (w/v) pelargonic acid in isopropanol solution and isopropanol vehicle. The Finn chambers were removed at 24 h post-application and reactions were scored according to the following scale: 0 (no visible reaction) to 4+ (intense erythema with bullous formation).

Pelargonic acid induced five 2+ (moderate erythema with edema and papules) reactions and seven 1+ (mild erythema with edema) reactions. The isopropanol vehicle induced five 1+ reactions and 5 +/- (faint, patchy erythema) reactions. Microscopic findings indicated that pelargonic acid disturbed the epidermal intercellular borders, which were described as blurred, thickened, and variably altered. These changes resulted from chemical damage to cellular membranes. The authors noted that pelargonic acid might increase the size of individual keratinocytes, interpreted as a result of intracellular disturbance with edema.97

Cetearyl Nonanoate The skin irritation potential of cetearyl nonanoate, 97% pure, was evaluated using 52 volunteers (males and females).45 The undiluted test substance (0.2 g) was applied to the upper back, and the application site was covered with an occlusive patch for 48 h. Reactions were scored at 48 h and 72 h post-application. Skin reactions were not observed in any of the subjects, and cetearyl nonanoate was classified as a non-irritant.

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Cetearyl Isononanoate A summary of a 1999 skin irritation study on Cetiol ® SN (cetearyl isononanoate, % not stated) was provided by the personal Care Products Council.64 Epicutaneous applications of the test substance (both 20% active substance and undiluted) were made to 21 volunteers over a 24 h period using Finn chambers (occlusive application). The test substance was rated as having very good skin compatibility.

Neopentyl Glycol Diisononanoate The skin irritation potential of undiluted neopentyl glycol diisononanoate was evaluated according to the preceding test procedure using 52 volunteers (males and females).50 Neopentyl glycol diisononanoate was classified as a non-irritant.

Ethyl Pelargonate According to Opdyke,37 skin irritation was not observed after ethyl pelargonate was applied (5 min to 5 h) to the skin of human subjects. At a concentration of 12% in petrolatum, ethyl pelargonate did not induce irritation in 48 h closed patch tests involving human subjects.

Smith et al.98 evaluated the skin irritation potential of ethyl pelargonate using 10 healthy volunteers (4 males, 6 females; 39 to 70 years old). The test substance (20% [w/w] in petrolatum) was applied to the upper back for 24 h using occlusive , 8 mm Finn chambers. The following served as controls: empty patch (negative control), petrolatum- water (vehicle control), and 20% sodium lauryl sulfate (positive control). Skin irritation was not observed in any of the subjects tested with ethyl pelargonate, and the same was true for negative and vehicle control test sites. The positive control was irritating to the skin of 5 subjects.

Provocative Tests In a study by Wahlberg and Maibach,99 75 patients with suspected allergic contact dermatitis (males and females; 20 to 70 years old) were patch tested with pelargonic acid at concentrations of 5.0 %, 10.0 %, 20 % (in propanol) and 100.0 % . Patches (Al-test® disc) were saturated with 0.04 ml of a test solution and applied to the upper back for 48 h. Reactions were scored at 48 h and 96 h post-application. At a concentration of 20%, skin irritation was observed in 93.9% of the 33 women tested and in 93.3% of the 48 men. All results from this study are summarized in Table 5.

Wahlberg et al.100 studied the skin irritation potential of pelargonic acid using 100 hospitalized patients with skin disease (54 males, 46 females; ages not stated). Pelargonic acid was tested at concentrations of 1.0, 5.0, 10.0, 20.0, and 39.9% in 1-propanol. Patches (Al-test ® discs) saturated with the test solutions were applied to the upper back and removed 48 h later. Reactions were scored at 1 h and 72 h post-removal. Results were reported as the lowest test concentration that produced a reaction (individual scores not reported), and are summarized as follows: 1.0% pelargonic acid (1 male; 2 females), 5.0% pelargonic acid (23 males; 9 females), 10.0% pelargonic acid (18 males; 22 females), 20.0% pelargonic acid (12 males; 13 females), and 39.9% pelargonic acid (all patients).

SKIN IRRITATION AND SENSITIZATION

Skin irritation and sensitization studies (normal subjects) on cetearyl nonanoate, ethylhexyl isononanoate, neopentyl glycol diisononanoate, and product formulations containing isodecyl isononanoate (51.35%) or isononyl isononanoate (3.552%) did not indicate a potential for irritation or sensitization.

Predictive Tests

Cetearyl Nonanoate The skin sensitization potential of cetearyl nonanoate (97% pure) was evaluated in an RIPT using 106 volunteers (males and females).45 The undiluted test substance (0.2 g) was applied to the upper back under an occlusive patch, for a total of 9 induction applications. Following a 2-week non-treatment period, each subject was challenged at

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a new test site. Reactions were scored at 24 h and 72 h post-application. Cetearyl nonanoate did not induce skin irritation nor allergic contact sensitization in any of the subjects.

Ethylhexyl Isononanoate In an RIPT,80 the skin sensitization potential of undiluted ethylhexyl isononanoate was evaluated according to the preceding test procedure using 52 volunteers. At the beginning of the test, mild skin irritation was observed in 10 subjects. It was concluded that ethylhexyl isononanoate did not indicate a potential for allergic contact sensitization.

Isodecyl Isononanoate The skin irritation and sensitization potential of a makeup product containing 51.35% isodecyl isononanoate was evaluated in an RIPT involving 101 normal subjects ( 18 to 65 years old).101 The 3-week, induction phase consisted of 24-h semiocclusive patch applications to the upper back (between the scapulae). Following a 2-week, nontreatment period, challenge patches were applied (24 h) to new sites on the back. Induction and challenge reactions were scored immediately after patch removal according to the following scale: 0 (no visible skin reaction) to 4+ (erythema and edema with vesiculation). Challenge reactions were also scored at 48 h and 72 h post-removal. Six subjects each had either a ± (barely perceptible erythema) or 1+ (mild erythema) reaction during induction. At challenge, one subject had a ± reaction at 48 h and 72 h and 2 subjects had a ± reaction at 72 h. It was concluded that the makeup product had no dermal irritation or sensitization potential.

Isononyl Isononanoate In a human repeated insult patch test (HRIPT),102 the skin irritation and sensitization potential of a lipstick containing 3.552% isononyl isononanoate was evaluated. The number of subjects at the beginning of the study was not stated; however, 53 subjects (ages not stated) satisfactorily completed the study. A modified Draize HRIPT procedure was used. Semi-occlusive patches containing the lipstick (0.2 g) were applied to the back for 24 h during induction and challenge phases. Challenge patch applications were made to previously untreated sites, and reactions were scored at 24 h and 72 h post-application. Skin reactivity was not observed in any of the 53 subjects during the study.

In another HRIPT (same procedure),103 the skin irritation and sensitization potential of another lipstick product containing 3.128% isononyl isononanoate was evaluated. The number of subjects at the beginning of the study was not stated; however, 97 subjects (ages not stated) satisfactorily completed the study. Skin reactivity was not observed in any of the 97 subjects during the study.

Neopentyl Glycol Diisononanoate The skin sensitization potential of undiluted neopentyl glycol diisononanoate was evaluated in an RIPT using 106 volunteers (males and females).50 The undiluted test substance (0.2 g) was applied to the upper back under an occlusive patch, for a total of 9 induction applications. Following a 2-week non-treatment period, each subject was challenged at a new test site. Reactions were scored at 24 h and 72 h post-application. Neopentyl glycol diisononanoate did not induce skin irritation nor allergic contact sensitization in any of the subjects.

SKIN SENSITIZATION

Pelargonic acid and its esters are not sensitizers, based on the results of predictive human skin sensitization studies. Negative results were reported for pelargonic acid (12%), ethyl pelargonate (12%), and the following other esters (in formulation): cetearyl isononanoate (1%), cholesteryl nonanoate (20.86%), isotridecyl isononanoate (4.3%), isodecyl isononanoate (2.6%), isononyl isononanoate (24.66%), and PEG-5 isononanoate (14.5%). Provocative test results for undiluted Cetiol ® SN (cetearyl isononanoate, % not stated)and isotridecyl alcohol (5%) were also negative.

Predictive Tests Pelargonic Acid According to Opdyke,37 pelargonic acid produced no sensitization reactions in 25 volunteers when tested at a concentration of 12% (in petrolatum) in a maximization test.

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Cetearyl Isononanoate The skin sensitization potential of a liquid makeup remover containing 1.5% cetearyl isononanoate was evaluated in a maximization test using 25 healthy adult volunteers (20 females, 5 males;18 to 65 years old).104 During induction, the test substance (0.05 ml) was applied, for 48 h, under a Webril cotton cloth patch (15 mm disc) secured with occlusive tape. Each induction application was preceded by a 24-h sodium lauryl sulfate (SLS) patch application at the same site. Applications were made to the upper outer arm. Following a 10-day non-treatment period, a single challenge application of the test substance (48 h) was made to a new site on the opposite upper arm of each subject. Each challenge application was preceded by a 1-h SLS patch application. Reactions were scored at 48h or 72 h according to the following scale: 0 (not sensitized) to 3 (strong sensitization). Contact allergy was not observed in any of the subjects.

Cholesteryl Nonanoate The skin sensitization potential of a lipstick containing 20.86% cholesteryl nonanoate was evaluated in a maximization test using 28 healthy adult volunteers (10 males, 18 females; 21 to 57 years old).105 The test procedure was identical to that stated in the preceding maximization test on a makeup remover containing ceteraryl isononanoate. Contact allergy was not observed in any of the subjects.

Isotridecyl Isononanoate In another study,106 the skin sensitization potential of a facial cream containing 4.3% isotridecyl isononanoate was evaluated in a maximization test using 28 healthy volunteers (8 males, 20 females; 19 to 63 years old). The test procedure was identical to that stated in the human maximization test on a makeup remover containing cetearyl isononanoate earlier in the report text. Contact allergy was not observed in any of the subjects.

Isodecyl Isononanoate The skin sensitization potential of a day cream containing 2.6% isodecyl isononanoate was evaluated in a maximization test using 26 healthy volunteers (4 males, 22 females; 26 to 65 years old).107 The test procedure was identical to that stated in the human maximization test on a makeup remover containing cetearyl isononanoate earlier in the report text. Contact allergy was not observed in any of the subjects.

Isononyl Isononanoate The skin sensitization potential of a cream eye shadow containing 24.66% isononyl isononanoate was evaluated in a maximization test using 26 healthy volunteers (6 males, 20 females; 18 to 65 years old).108 The test procedure was identical to that stated in the human maximization test on a makeup remover containing cetearyl isononanoate earlier in the report text. Contact allergy was not observed in any of the subjects.

PEG-5 Isononanoate The skin sensitization potential of an experimental formulation with the following composition was evaluated in an RIPT using 53 volunteers: PEG-5 isononanoate (14.5%), water, trideceth-9, sodium hydroxide, and carbomer.51 Concentrations of the remaining components were not stated. The formulation (0.2 g) was applied, under an occlusive patch, to the upper back of each subject for a total of 9 induction applications. After a 2-week non-treatment period, a challenge patch was applied to a new site. Reactions were scored at 24 h and 72 h post-application. Reactions were not observed during the study, and it was concluded that the formulation did not indicate a potential for allergic contact sensitization.

Ethyl Pelargonate According to Opdyke,37 results for 12% ethyl pelargonate in petrolatum were negative in 25 volunteers evaluated in the maximization test.

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Provocative Tests

Cetearyl Isononanoate The skin sensitization potential of undiluted Cetiol ® SN (cetearyl isononanoate, % not stated) was evaluated in a repeated insult patch test using 20 male patients with eczema and other skin diseases.109 The test substance was applied, under Beiersdorf test plaster, at 2-day intervals for a total of 10, 24 h applications. The patients were challenged, following a 14-day non-treatment period. Cetiol ® SN did not induce skin irritation or sensitization. Isotridecyl Alcohol Geier et al.110 evaluated the sensitization potential of isotridecyl alcohol (5% in petrolatum) using 229 dermatitis patients (mean age = 39.3 years). The patch application time was 1 or 2 days. All tests were read at least until day 3. The test substance did not induce sensitization in any of the subjects tested.

EPIDERMAL PROLIFERATION AND APOPTOSIS

Pelargonic acid (80% in propanol) increased the density of proliferating keratinocytes and caused epidermal cell apoptosis and a transient decrease in keratinocyte proliferation.

Willis et al.111 investigated alterations in the proliferative capacity of human epidermis following topical exposure to pelargonic acid (80% [v/v] in propan-1-ol). Finn chambers containing the test substance were applied to the volar aspect of the forearm of each of 10 healthy, non-atopic male volunteers for 48 h. Punch biopsies were removed from each application site. Samples of normal skin were also obtained. Compared to the vehicle control, pelargonic acid induced a statistically significant (p < 0.05) increase in the density of proliferating keratinocytes (i.e., increase in mitotic activity).

Forsey et al.112 studied the effect of 80% pelargonic acid (in propan-1-ol) on Langerhans cells and on epidermal proliferation and apoptosis. Punch biopsies were obtained from the volar forearm of 46 irritant contact dermatitis patients (25 males, 21 females; age range = 18 to 62) and 10 healthy subjects, following application of the test substance (Finn chambers) and vehicle control to the skin for up to 48 h. A higher number of Langerhans cells/mm basement membrane in the patients, compared to controls, was reported. However, there was no difference in the number of dendrites/Langerhans cell or in dendrite length. Pelargonic acid caused a decrease in keratinocyte proliferation after 24 h of exposure, but a return to basal levels was observed after 48 h. Pelargonic acid induced epidermal cell apoptosis after only 6 h of exposure, and dramatically decreased the Langerhans cell number after 24 h and 48 h of exposure. Apoptosis was induced in over half of the Langerhans cells that were present after 24 h and 48 h.

EFFECT ON CD1a AND ICAM-1 EXPRESSION

Lindberg et al.113 investigated the possibility of differences in the interaction between different irritants (pelargonic acid and SLS) and immunological parameters in the epidermis using 9 healthy volunteers. The reactions were evaluated by immunohistochemistry using monoclonal antibodies directed against CD1a, CD3, and intercellular adhesion molecule-1 (ICAM-1) molecules. Initially, occlusive patch tests (Finn chambers) involving the following 3 groups were conducted. In group 1 (2 males, 1 female; ages: 26 to 52 years), 2% SLS and 4% SLS in distilled water (w/v) and distilled water alone were applied under occlusion for 24 h, and biopsies were obtained at 48 h. In group 2 (3 males; ages: 23 to 37 years), 20% pelargonic acid and 80% pelargonic acid in isopropanol (v/v), and isopropanol alone were applied under occlusion for 24 h, and biopsies were obtained at 48 h. In group 3 (3 males; ages 24 to 25 years), 4% SLS in water and 80% pelargonic acid in isopropanol were applied under occlusion for 24 h; biopsies were obtained immediately after Finn chamber removal.

At 48 h (groups 1 and 2), marked edema was observed at the 4% SLS site; this reaction was greater in severity when compared to those induced by 20% and 80% pelargonic acid. At 24 h (group 3), the reactions to 4% SLS and 80% pelargonic acid were similar. Reactions were not induced by the distilled water or isopropanol vehicle. At both 24 h and 48 h, an exposure-related (SLS and pelargonic acid) increase in the number of CD3+ cells in the upper part of the dermis was observed. A minor increase in CD3+ cells following exposure to distilled water, but not isopropanol,

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was also observed at 48h. Few CD3+ cells were also identified in the epidermis following SLS and pelargonic acid exposure.

Differences in ICAM-1 expression in the epidermis following SLS and pelargonic acid exposure were observed. Following SLS exposure, an increase in ICAM-1+ keratinocytes at 24 h and 48 h was noted. A tendency toward increased numbers of CD1a+ cells was noted at 48 h after treatment with 4% SLS. A definite decrease in the number of CD1a+ cells was observed following exposure to 80% pelargonic acid. ICAM-1-reactivity was not detected in the epidermis following exposure to 20% or 80% pelargonic acid. Increased levels of ICAM-1 expression were observed in the epidermis following exposure to both water and isopropanol controls. In the control biopsies, 3 of 9 specimens had ICAM-1 reactivity (single cells or few keratinocytes). It was concluded that different irritants applied to the skin surface may induce different responses in epidermis measured with markers for immunological components, although the clinical picture indicates primary irritancy.113

CASE REPORTS Pelargonic Acid Mathias et al.114 were authors of a case report on perioral leukoderma, simulating vitiligo, from the use of a toothpaste containing cinnamic aldehyde. Pelargonic acid (10% and 20% aqueous solutions) was used as a positive irritant control in patch tests that were performed on the 25-year-old female subject. Occlusive patches were applied for 48 h, and reactions were scored at 48 h and 96 h according to the method of the North American Contact Dermatitis Group. Both solutions of pelargonic acid produced erythema and slight induration, confined to patch test sites. Hyperpigmentation was observed at the sites of irritant reactions to pelargonic acid.

Cetearyl Isononanoate In a case report by Le Coz and Bressieux,115 a 23-year-old female with a history of allergic contact dermatitis developed acute dermatitis after application of a urea-based moisturizing cream containing cetearyl isononanoate. The subject was patch tested (repeated open application test) with ingredients of the cream, each diluted to a concentration identical to that in the product. Allergic reactions to the cream (day 2: ++; day 4: ++) and cetearyl isononanoate (day 2: negative; day 4: ++), both diluted to concentration of 4% in liquid mineral oil were reported. Patch test results for 4% cetearyl isononanoate were negative in 10 voluntary control subjects on days 2, 3, and 4.

Isononyl Isononanoate In a case report by Goossens et al.,116 a 40-year-old, non-atopic female presented with contact cheilitis following application of a lipstick product containing isononyl isononanoate in 2002. In 2007, she presented with severe contact dermatitis on the eyelids following application of a new lipstick product containing isononyl isononanoate to the eyelids. Patch test results for isononyl isononanoate were positive. Additional patch testing (patch test chambers on forearm) was performed, and reactions were scored according to International Contact Dermatitis Research Group criteria on days 2 and 4, and later. Strong (vesicular) positive reactions were observed at all tested ethanolic dilutions of isononyl isononanoate, from 20% (actual use concentration in product) to 1%. The patient developed severe edema of the entire test site. Patch test results for isononyl isononanoate (5% in ethanol) were negative in 20 control subjects. Further patch testing in 2008 to identify possible cross-reactions did not yield further positive reactions.

SUMMARY OF INFORMATION FROM EARLIER CIR SAFETY ASSESSMENTS

The CIR Final Safety Assessment on propylene glycol esters and diesters included propylene glycol dipelargonate with a safe as used conclusion, but with a concern in the discussion regarding penetration enhancement with propylene glycol dipelargonate. Butyl, cetyl, isobutyl, isocetyl, isopropyl, myristyl, and octyl stearate (now ethylhexyl stearate) were reviewed as a group of fatty acid esters and found safe as used. Decyl and isodecyl oleate were reviewed together and found safe as used, based in part on the chemical similarity of the compounds and the corresponding similarities in the available animal and human safety test data.

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Propylene Glycol Esters and Diesters – including Propylene Glycol Dipelargonate

There were limited data on many of the propylene glycol esters and diesters. There were data indicating that propylene glycol dicaprylate/dicaprate was a minimal dermal irritant and was not comedogenic. In the discussion, it was noted that the caprylic (8 carbon chain)/capric (10 carbon chain) moiety is similar to the dipelargonate (9 carbon chain) moiety. Propylene glycol dipelargonate enhanced the skin penetration of caffeine and testosterone through human stratum corneum in vitro.

Overall, the Panel relied substantially on the prior reviews of the following ingredients previously reviewed to demonstrate overall safety of this group:  propylene stearate for mutagenicity, chronic toxicity and skin sensitization;  caprylic/capric triglyceride for reproductive toxicity, chronic toxicity, and skin sensitization  coconut acid for chronic toxicity, tumor promotion, skin sensitization, phototoxicity, and photosensitization  isostearic acid for skin sensitization, photosensitization, and phototoxicity  lauric, myristic, and oleic acids for reproductive toxicity, carcinogenicity, skin sensitization, and photosensitization;

In the original safety assessment of isostearic acid, two reports that discussed metabolism were noted. One study concluded that rat liver homogenate acyl coenzyme A synthetase was found to activate isostearic acid. In another study that focused specifically on metabolism of iso-fatty acids versus straight-chain fatty acids, it was reported that metabolism is similar by the mitochondrial and microsomal fractions of rat liver homogenate. The straight-chain fatty acids are successively oxidized at the β carbon to yield two carbon fractions. The iso-fatty acids also follow that path, but, in addition, are oxidized at the ω carbon to ultimately form three carbon dicarboxylic acids. The enzymes catalyzing the ω-hydroxylation are present in the mitochondrial and microsomal fractions, whereas the enzymes catalyzing further oxidation into carboxylic acids are in the soluble fractions of rat liver homogenate.

In the discussion, the Panel did note a concern about dermal penetration enhancement with propylene glycol dipelargonate. It has been shown that propylene glycol dipelargonate enhances the skin penetration of caffeine through human stratum corneum in vitro.

Butyl, Cetyl, Isobutyl, Isocetyl, Isopropyl, Myristyl, and Octyl (now Ethyhexyl) Stearate

Few data were available that demonstrated the metabolic fate of the iso forms compared to straight chain forms beyond noting that aliphatic esters are hydrolyzed to the corresponding alcohol and fatty acid and further metabolized. 3 One study was provided in which isopropyl stearate, diluted in 9,10- H2-labeled oleic acid, was given by gavage to thoracic duct fistula rats. Radiolabel was found in lymph lipids, suggesting a dietary origin. Less than 10% of the recovered radiolabel was in the form of the isopropyl ester and, conversely, over 95% of the radiolabel was in triglycerides, leading to the suggestion that the isopropyl ester is hydrolyzed in the intestine and that the fatty acids thus liberated are reesterified before distribution to lymph lipids.

Safety test data indicated low acute oral toxicity, no reproductive toxicity, and minimal skin irritation, but no skin sensitization, phototoxicity, or photosensitization. The discussion noted the absence of data on comedogenicity.

Decyl and Isodecyl Oleate

These fatty acid esters have low acute oral toxicities, but few other toxicity data were available. They can be minimal to moderate dermal irritants, but are not sensitizers in human and animal tests. No metabolic fate information relevant to the esters was available.

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SUMMARY

Pelargonic acid and nonanoate esters are cosmetic ingredients that function as skin conditioning agents in cosmetics. The following ingredients are reported as being used: butylene glycol diisononanoate, cetearyl isononanoate, cetearyl nonanoate, cetyl isononanoate, cholesteryl nonanoate, diethylene glycol diethylhexanoate/diisononanoate, dipentaerythrityl pentaisononanoate, ethylhexyl isononanoate, isodecyl isononanoate, isononyl isononanoate, isotridecyl isononanoate, tridecyl isononanoate, ethylhexyl pelargonate, neopentyl glycol diisononanoate, cetearyl nonanoate, cetyl isononanoate, dipentaerythrityl pentaisononanoate, neopentyl glycol diisononanoate, PEG-2 diisononanoate, PEG-5 isononanoate, pentaerythrityl tetraisononanaote, polyglyceryl-20 octaisononanoate, and pentaerythrityl tetrapelargonate. Current ingredient use concentrations range from 0.01% (cholesteryl nonanoate) to 74% (ethylhexyl isononanoate).

The following chemicals do not absorb significantly in the 250 to 400 nm range: neopentyl glycol diisononanoate, cetyl nonanoate + stearyl nonanoate, trideceth-9 + PEG-5-isononanoate + water, glyceryl triisononnoate + glyceryl diisononanoate, and ethylhexyl isononanoate.

Straight-chain pelargonic acid esters are likely hydrolyzed to component alcohols and pelargonic acid, which is further metabolized by β-oxidation. Iso-fatty acids and straight-chain fatty acids both are metabolized at the β-carbon to yield two-carbon fractions by mitochondrial and microsomal fractions of rat liver homogenate. Additionally, iso- fatty acids are oxidized at the ω carbon to ultimately form three-carbon dicarboxylic acids. The enzymes catalyzing the ω-hydroxylation are present in the mitochondrial and microsomal fractions, whereas the enzymes catalyzing further oxidation into carboxylic acids are in the soluble fractions of rat liver homogenate. With the exception of pelargonic acid and ethyl pelargonate, specific information relating to the metabolism of the remaining ingredients reviewed in this safety assessment was not identified in the published literature. Branched-chain fatty acid metabolism involves initial α-oxidation, which is followed by the β-oxidation pathway.

Octanol-water partition coefficient (logP) and molecular weight data included in the safety assessment may be used to predict the skin penetration potential of pelargonic acid and its esters/ester moieties. Most of the ingredients reviewed in this safety assessment have a logP of > 5 and a mw of < 500. Compounds with a logP of > 5 and a mw of > 500 are less likely to penetrate the skin. For example, cholesteryl nonanoate has a logP of 10 and a mw of > 500, suggesting that dermal absorption is unlikely. The skin penetration enhancement effect of pelargonic acid on other chemicals has been demonstrated in vitro using human stratum corneum and hairless rat skin. The percutaneous absorption of isononyl alcohol was reported in an acute dermal toxicity study using rabbits with abraded skin and occlusion of the site.

Pelargonic acid and the esters for which data are available are not significant acute oral toxicants (LD50s > 1 g/kg) or acute dermal toxicants (lowest LD50 reported = 5 g/kg).

Short-term oral dosing with pelargonic acid (in diet) yielded LOELs of 5,000 ppm (for clinical pathology) and 100 ppm (for antemortem data) in rats. Overt signs of toxicity were not associated with higher doses in other short-term oral toxicity studies. Short-term oral dosing with isononyl isononanoate induced liver and kidney toxicity in rats at doses up to 1,000 mg/kg/day. Limited numbers of rats given isononyl alcohol at 1 mmol/kg/day had no overt signs of toxicity. The NOAEL for neopentyl glycol in rats was 100 mg/kg. Subchronic oral dosing with 1% ethyl pelargonate (in diet) did not result in any remarkable gross or microscopic findings in rats. However, subchronic oral dosing with Cetiol ® SN (cetearyl isononanoate, % not stated) induced reversible fatty alterations in the liver of rats, and an NOAEL of 100 mg/kg/day was reported.

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studies, dermal application of pelargonic acid to mice over a 3-day period did not cause death and a TDLo of 3,000 mg/kg was reported. Short-term cutaneous dosing with isononyl isononanoate induced liver and adrenal toxicity in rats at doses up to 860 mg/kg/day.

The ocular changes observed in a 28-day oral toxicity study (rats) on pelargonic acid were considered sporadic and unrelated to treatment. In studies involving rabbits, pelargonic acid (0.1 ml) was severely irritating or mildly irritating. Neither isononyl isononanoate, ethylhexyl pelargonate (0.1 ml), nor Cetiol ® SN (cetearyl isononanoate, % not stated) at a concentration of 10% (0.5 ml) induced ocular irritation in rabbits; however, both cetearyl nonanoate and neopentyl glycol diisononanoate (0.1 ml) induced minimal ocular irritation and isononyl alcohol (0.1 ml) induced marked ocular irritation. PEG-5 isononanoate (0.1 ml) induced transient ocular reactions, but was not classified as an ocular irritant.

Undiluted pelargonic acid was a mild to severe skin irritant in rabbits and a severe skin irritant in guinea pigs, but was not irritating to the skin of mice. However, undiluted cetearyl isononanoate was non-irritating to the skin of rabbits, but, as undiluted Cetiol ® SN (cetearyl isononanoate, % not stated), was slightly irritating to the skin of mice. The remaining studies involved rabbits only. Isononyl isononanoate (unknown concentration) and undiluted PEG-5 isononanoate were slightly/mildly irritating and undiluted ethyl pelargonate was moderately irritating to the skin; however, undiluted isononyl alcohol induced marked skin irritation. Skin irritation was not observed following the application of undiluted cetearyl nonanoate, cetearyl isononanoate, or undiluted ethylhexyl pelargonate. Repeated applications of Cetiol ® SN (cetearyl isononanoate, % not stated) to the rabbit ear at concentrations ranging from 10% to 100% did not cause any alterations or produce structures typical of comedogenicity in the infrainfundibulum of hair follicles.

In an RIPT, pelargonic acid (50% in corn oil) induced skin irritation, but not sensitization, in guinea pigs. Sensitization test results for guinea pigs injected intracutaneously with Cetiol ® SN (cetearyl isononanoate, % not stated) at a concentration of 25% did not differ from those of control guinea pigs. In guinea pig maximization tests, cetearyl nonanoate (10% in sesame oil) and undiluted neopentyl glycol diisononanoate were non-sensitizers; results for 50% cetearyl nonanoate in sesame oil and undiluted neopentyl glycol diisononanoate (for induction) were negative in preliminary skin irritation tests. Both the methyl ester and propyl ester of pelargonic acid (both at 0.2% and 2.0%) were said to have shown a sensitization tendency in a study examining the tissue response to pelargonic acid in the buccal mucosa of the rat. In mouse LLNAs (for sensitization potential), results were positive at pelargonic acid concentrations of ≥ 50% and ≥ 20% (no-effect-level = 10%) and negative for PEG-5 isononanoate at concentrations up to 100%.

Daily doses of pelargonic acid up to 1,500 mg/kg/day did not induce reproductive effects in inseminated female rats. Results from other studies support pelargonic acid daily doses of 1500 mg/kg/day as the no-observable- effect-level for maternal/developmental toxicity in rats, and isononyl isononanoate daily doses of 300 mg/kg/day as the NOEL for developmental toxicity in rats. In a teratogenicity study on Cetiol ® SN (cetearyl isononanoate, % not stated), the NOAEL for maternal toxicity and embryo-/fetotoxicity was 1000 mg/kg body weight. Two branched-chain nonanols (perhaps incorrectly identified as isononanol) caused a marked degree of maternal and fetal toxicity in rats at daily doses of 7.5 and 10 mmol/kg/day and slight fetal effects at 5 mg/kg/day doses. Neopentyl glycol at oral doses up to 1,000 mg/kg/day did not induce reproductive effects.

While one mammalian cell assay of pelargonic acid was positive with metabolic activation (doses up to 600 µg/ml), it was negative without metabolic activation. All other bacterial, mammalian cell, and in vivo assays for pelargonic acid (doses up to 5,000 µg/plate or 5,000 mg/kg) and the following pelargonic acid esters (doses up to 5,000 µg/plate) were negative: Cetiol ® SN (cetearyl isononanoate, % not stated), cetearyl nonanoate, ethylhexyl isononanoate, isononyl isononanoate, neopentyl glycol diisononanoate, and PEG-5 isononanoate. Negative results were also reported for neopentyl glycol at doses up to 5,000 µg/plate (bacterial cells)and up to 5 mg/ml (mammalian cells). There was no evidence of gross skin tumors in mice dosed with undiluted pelargonic acid in a dermal carcinogenicity study.

Pelargonic acid is a known skin irritant, based on and the results of both predictive and provocative human skin irritation studies. In predictive tests, pelargonic acid induced skin irritation at concentrations ranging from 5% to 80%; ethyl pelargonate was a skin irritant at a concentration of 20%, but not 12%. Predictive human skin irritation test 34

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results for undiluted cetearyl nonanoate, Cetiol ® SN (cetearyl isononanoate, % not stated) at a concentration of 20%, and undiluted neopentyl glycol diisononanoate were negative, and the same was true for predictive human skin irritation and sensitization studies on cetearyl nonanoate, ethylhexyl isononanoate, and neopentyl glycol diisononanoate, all undiluted, and product formulations containing isodecyl isononanoate (51.35%) and isononyl isononanoate (3.552%). Similarly, predictive human skin sensitization studies on 12% pelargonic acid, 12% ethyl pelargonate, and formulations containing the following pelargonic acid esters were negative: cetearyl isononanoate (1.5%), cholesteryl nonanoate (20.86%), isotridecyl isononanoate (4.3%), isodecyl isononanoate (2.6%), isononyl isononanoate (24.66%), and PEG-5 isononanoate (14.5%). Results were negative for undiluted Cetiol ® SN (cetearyl isononanoate, % not stated) and 5% isotridecyl alcohol in provocative human skin sensitization studies.

In other human studies, pelargonic acid (80%) increased the density of proliferating keratinocytes and caused epidermal cell apoptosis and a transient decrease in keratinocyte proliferation.

Patch testing with Pelargonic acid caused skin irritation and hyperpigmentation in a single case report. In other case reports, a moisturizing cream containing cetearyl isononanoate induced contact dermatitis and a lipstick containing 20% isononyl isononanoate induced contact cheilitis. Patch test results for 4% cetearyl isononanoate and 5% isononyl isononanoate were negative in healthy control subjects in these reports.

DISCUSSION

The CIR Expert Panel recognizes that the pelargonic acid branched esters included in this safety assessment are not pure substances, but are always mixtures. Thus, the international nomenclature cosmetic ingredient (INCI) name for each pelargonic acid branched ester actually refers to a mixture with general properties that, in many instances, cannot be fully characterized. Regardless, the material, as supplied to the industry for use in cosmetics, is the same material that is used in safety testing. While specific data on the metabolic fate of the branched esters included in this safety assessment were not found, data on phytanic acid, a 20-carbon branched fatty acid, indicate that the α-methylene group is oxidatively excised (α-oxidation) in mammalian microsomes, yielding pristanic acid. Pristanic acid is then metabolized via the β-oxidation pathway. These data were deemed representative of the metabolic pathways for branched fatty acids in general.

The Expert Panel noted the availability of acute inhalation toxicity data on pelargonic acid, isononanoic acid, and isononyl alcohol, but not on any other of the ingredients that are being reviewed in this safety assessment. However, in the absence of these data, the Panel determined that the ingredients included in this review can be used safely in hair sprays, because the product particle size is not respirable. The Panel reasoned that the particle size of aerosol hair sprays (around 38 µm) and pump hair sprays (>80 µm) is large compared to respirable particle sizes (≤10 µm). The Panel expressed concern over the findings of liver steatosis and acidophilic globules in the renal cortical tubules of male rats in both short-term oral and dermal toxicity studies on isononyl isononanoate, but also acknowledged industry comments to the effect that high-fat diets produce liver steatosis. This sex- and species-specific hyaline droplet nephropathy was not considered relevant to man because the alpha-2-microglobulin protein is absent from man as well as many species.

Because animal sources of cetearyl isononanoate, cetyl isononanoate, cholesteryl nonanoate, dihydrocholesteryl nonanoate, and isostearyl isononanoate have been reported, the Panel was also concerned with the dangers inherent in using animal-derived ingredients, namely the transmission of infectious agents. The CIR Expert Panel stressed that the preceding ingredients must be free of detectable pathogenic viruses or infectious agents. Suppliers and users of these ingredients must accept responsibility for assuring that these ingredients are risk-free. Tests to assure the absence of a pathogenic agent in the ingredients, or controls to assure derivation from pathogen-free sources are two approaches that should be considered.

The Expert Panel also recognized that pelargonic acid and related ingredients, because of the skin penetration enhancement property of pelargonic acid in the presence of p-aminobenzoic acid, could enhance the penetration of similar chemicals, possibly cosmetic ingredients, through the skin. The Panel cautioned that care should be taken in

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formulating cosmetic products that may contain these ingredients in combination with any ingredients whose safety was based on their lack of dermal absorption data, or when dermal absorption was a concern.

Panel deliberations on the safety of pelargonic acid esters focused on current ingredient use concentration data from industry in relation to human RIPT data on cosmetic products containing these ingredients. Current ingredient use concentrations ranged from 0.01% (cholesteryl nonanoate) to 74% (ethylhexyl isononanoate), and ethylhexyl isononanoate was a mild skin irritant, but a nonsensitizer, when tested undiluted. Similarly, cetearyl isononanoate, cetearyl nonanoate, and neopentyl glycol diisononanoate, each undiluted, did not induce sensitization in human RIPTs. Other data indicate that the highest ester concentration, undiluted excluded, evaluated in a human RIPT was 51.35% isodecyl isononanoate in a makeup product (nonirritant and nonsensitizer), which approaches the maximum use concentration of 59% for this ingredient. Results from a non-good laboratory practice (GLP) sensitization study on Cetiol ® SN (cetearyl isononanoate, % not stated) at a test concentration of 25% were also considered only because the skin reactions of test animals did not differ from those of controls. Based on negative results for sensitization at high ingredient test concentrations, the Expert Panel reasoned that is not likely that the pelargonic acid esters reviewed in this safety assessment would induce sensitization in the present practices of use.

CONCLUSION The CIR Expert Panel concluded that butylene glycol diisononanoate, cetearyl isononanoate, cetearyl nonanoate, cetyl isononanoate, cholesteryl nonanoate, diethylene glycol diethyl-hexanoate/diisononanoate, dipentaerythrityl pentaisononanoate, ethylhexyl isononanoate, isodecyl isononanoate, isononyl isononanoate, isotridecyl isononanoate, neopentyl glycol diisononanoate, PEG-2 diisononanoate, PEG-5 isononanoate, pentaerythrityl tetraisononanaote, polyglyceryl-20 octaisononanoate, tridecyl isononanoate, ethylhexyl pelargonate, pentaerythrityl tetrapelargonate, cellobiose octanonanoate, diethylene glycol diisononanoate, dihydrocholesteryl nonanoate, glycereth-7 diisononanoate, isostearyl isononanoate, phytosteryl nonanoate, propylene glycol diisononanoate, ethyl pelargonate, isobutyl pelargonate, methyl pelargonate, neopentyl glycol dicaprylate/dipelargonate/dicaprate, and pelargonic acid are safe as cosmetic ingredients in the present practices of use and concentration described in this safety assessment.1

1 Were ingredients in this group not in current use to be used in the future, the expectation is that they would be used in product categories and at concentrations comparable to others in the group. 36

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Table 1. Pelargonic Acid and its Nonanoate Esters15 Chemical Names Definition Functions in Cosmetics pelargonic acid; nonanoic acid; nonoic acid; an acid that conforms to the formula in fragrance ingredients; surfactants – nonylic acid; 1-octanecarboxylic acid; pelargic figure 1 cleansing agents; surfactants – acid; pergonic acid; CAS No. 112-05-0 emulsifying agents butylene glycol diisononanoate; trade name: the diester of butylene glycol and skin-conditioning agents – emollient; cetiol FC branched chain nonanoic acids skin-conditioning agents – occlusive; viscosity increasing agents – nonaqueous cellobiose octanonanoate; α-D-glycopyranose, the octaester formed by the reaction of viscosity increasing agents – 4-O-[2,3,4,6-tetrakis-O-(1-oxononyl)-β-D- α-D-cellobiose and nonanoic anhydride nonaqueous glucopyranosyl]- ,tetranonanoate; CAS No. 172585-66-9; trade name: alpha-D-cellobiose octanonanoate cetearyl isononanoate; isononanoic acid, the ester of cetearyl alcohol and a hair conditioning agents; skin- cetyl/stearyl ether; tradenames: AEC cetearyl branched chain nonanoic acid conditioning agents – emollient isononanoate; cetiol SN; dub IN 1618; saboderm CSN; and tegosoft CI cetearyl nonanoate; trade name: SymMollient S the organic compound that conforms to skin-conditioning agents – emollient 181598 the formula in figure 1, where R represents the cetearyl group cetyl isononanoate; isononanoic acid, hexadecyl the ester of cetyl alcohol with a skin-conditioning agents – emollient ester; CAS No. 84878-33-1 branched chain nonanoic acid cholesteryl nonanoate; cholesterin pelargonate; the ester of cholesterol and nonanoic not reported cholesteryl nonylate; cholesteryl pelargonate; acid CAS No. 1182-66-7; trade name: yofco LC-CN diethylene glycol the diester of a mixture of 2- Hair conditioning agents; plasticizers; diethylhexanoate/diisononanoate ethylhexanoic acid and isononanoic skin-conditioning agents – emollient acids and diethylene glycol diethylene glycol diisononanoate; isononanoic the diester of diethylene glycol and hair conditioning agents; plasticizers; acid, oxydi-2,1-ethanediyl ester; PEG-2 isononanoic acid skin-conditioning agents – emollient diisononanoate; CAS Nos. 106-01-4;190282-37- 2 dihydrocholesteryl nonanoate the ester of dihydrocholesterol and skin-conditioning agents – emollient nonanoic acid dipentaerythrityl pentaisononanoate; CAS the pentaester of isononanoic acid with skin-conditioning agents – emollient; No. 84418-63-3; trade name: dub vinyl a dimer of pentaerythritol viscosity increasing agents – nonaqueous ethylhexyl isononanoate; 2-ethyl-hexyl the ester of 2-ethyl-hexyl alcohol and a skin-conditioning agents – emollient isononanoate; 2-ethylhexyl isopelargonate; 2- branched chain nonanoic acid ethylhexyl 3,5,5-trimethyl-hexanoate; isononanoic Acid, 2-ethylhexyl ester; octyl isononanoate [sic], former INCI ingredient name; CAS Nos.70969-70-9; 71566-49-9; trade names: AEC ethylhexyl isononanoate; dermol 89; dub INO; ES 108109; HallStar octyl isononanoate; isolanoate; pelemol 89; schercemol OISN glycereth-7 diisononanoate; PEG-7 glyceryl the diester of isononanoic acid and skin-conditioning agents – emollient; ether diisononanoate; poly-ethylene glycol (7) glycereth-7 solvents glyceryl ether diisononanoate; polyoxyethylene (7) glyceryl ether diisononanoate; CAS No. 125804-15-1; trade name: dermol G-7DI

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Table 1. Pelargonic Acid and its Nonanoate Esters15 Chemical Names Definition Functions in Cosmetics isodecyl isononanoate; hexanoic acid, 3,5,5,- the ester of branched chain decyl skin-conditioning agents – emollient trimethyl-, isodecyl ester; isodecyl 3,5,5- alcohols and branched chain nonanoic trimethylhexanoate; isononanoic acid, isodecyl acid that conforms to the structure in ester; 3,5,5-trimethylhexanoic acid, isodecyl figure 1 ester; CAS Nos. 41395-89-5 and 59231-35-5; trade names: AEC isodecyl isononanoate; dermol 109; dub INID; KAK 109 isononyl isononanoate; hexanoic acid, 3,5,5- the ester of a branched chain nonyl skin-conditioning agents – emollient trimethyl-, 3,5,5-tri-methylhexyl ester; alcohol with a branched chain nonanoic isononanoic acid, isononyl ester; 3,5,5- acid trimethylhexa-noic acid, 3,5,5-trimethylhexyl ester; 3,5,5-trimethylhexyl-3,5,5-trimethylhexanoate; CAS Nos. 42131-25-9 and 59219- 71-5; trade names: AEC isononyl isononanoate; dermol 99; dub ININ; hatcol 5131; KAK 99; lanol 99; pelemol IN-2; saboderm ISN; salacos 99 isostearyl isononanoate; hexanoic acid, 3,5,5- the ester of isostearyl alcohol and skin-conditioning agents – emollient trimethyl-, isooctadecyl ester; CAS Nos. 90967- isononanoic acid 66-1and 163564-45-2; trade name: lanol 189 isotridecyl isononanoate; hexanoic acid, 3,5,5- the ester of isotridecyl alcohol and skin-conditioning agents – emollient trimethyl-, isooctadecyl ester; isononanoic acid, isonona-noic acid isotridecyl ester; isotridecyl 3,5,5-trimethylhexanoic acid; CAS Nos. 42131-27-1 and 59231-37- 7; trade names: AEC isotridecyl isononanoate; dub INI; KAK 129; OriStar ITIN; salacos 913 neopentyl glycol diisononanoate; CAS No. the organic compound that conforms to skin-conditioning agents – emollient 137636-04-5; trade names: NPDIN; the structure in figure 1 SymMollient L 177205

PEG-2 diisononanoate; polyethylene glycol the polyethylene glycol diester of surfactants – emulsifying agents 100 diisononanoate; polyoxy-ethylene (2) isononanoic acid that conforms to the diisononanoate; structure in figure 1, where n has an average value of 2 PEG-5 isononanoate the organic compound that conforms surfactants – emulsifying agents generally to the structure in figure 1, where n has an average value of 5 pentaerythrityl tetraisononanoate; 2,2-bis[[(1- the tetraester of pentaerythritol and a skin-conditioning agents – occlusive; oxoisononyl)oxy]methyl] branched chain nonanoic acid. It viscosity increasing agents – -1,3-propanediyl isononanoate; isononanoic conforms generally to the structure in nonaqueous acid, 2,2-bis[[(1-oxoisononyl)oxy]methyl]-1,3- figure 1, where RCO- represents the propanediyl ester; CAS No. 93803-89-5; trade isononanoic acid radical name: pelemol P-49

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Table 1. Pelargonic Acid and its Nonanoate Esters15 Chemical Names Definition Functions in Cosmetics phytosteryl nonanoate; trade name: yofco LC- the ester of phyto-sterol and nonanoic hair conditioning agents; skin- PN acid conditioning agents – miscellaneous polyglyceryl-20 octaisononanoate; trade name: the octaester of isononanoic acid and surfactants – cleansing agents; sunsoft Q-98U polyglycerin-20 surfactants – emulsifying agents; surfactants – solubilizing agents propylene glycol diisononanoate; isononanoic the diester of propylene glycol and skin-conditioning agents – occlusive; acid, 1-methyl-1,2-ethanediyl ester; CAS No. branched chain nonanoic acids viscosity increasing agents – 125804-17-3; trade names: AEC propylene nonaqueous glycol diisononanoate; dermol PGDI tridecyl isononanoate; hexanoic acid, 3,5,5- the ester of tridecyl alcohol and skin-conditioning agents – emollient trimethyl-, tridecyl ester; isononanoic acid that conforms to the isononanoic acid, tridecyl ester; structure in figure 1 3,5,5-trimethylhexanoic acid, tridecyl ester; CAS No. 125804-18-4; trade name: dermol 139 ethylhexyl pelargonate; 2-ethylhexyl ester of 2-ethylhexyl alcohol and Skin-conditioning agents – emollient pelargonate; nonanoic acid, w-ethylhexyl ester; pelargonic acid that conforms to the octyl pelargonate; CAS No. 59587-44-9; trade structure in figure 1 names: AEC ethylhexyl pelargonate; bernel ester OPG; crodamol OPG; dub PEO; jeechem OPG; pelemol OPG; schercemol OPG ester ethyl pelargonate; ethyl nonanoate; nonanoic ester of ethyl alcohol and pelargonic fragrance ingredients; hair conditioning acid, ethyl ester; CAS No. 123-29-5 acid that conforms to the structure in agents; skin-conditioning agents – figure 1 emollient isobutyl pelargonate; isobutyl nonanoate; 2- ester of isobutyl alcohol and pelargonic fragrance ingredients; skin-conditioning methylpropyl nonanoate; nonanoic acid, isobutyl acid that conforms to the structure in agents – emollient ester; nonanoic acid, 2-methypropyl ester; CAS figure 1 No. 30982-03-7; trade name: AEC isobutyl pelargonate methyl pelargonate; methyl nonanoate; ester of methyl alcohol and pelargonic fragrance ingredients; skin-conditioning nonanoic acid, methyl ester; pelargonic acid acid that conforms to the structure in agents – emollient methyl ester; CAS No. 1731-84-6 figure 1 neopentyl glycol diester of neopentyl glycol and a blend skin-conditioning agents – emollient; dicaprylate/dipelargonate/dicaprate of caprylic, pelargonic, and capric acids viscosity increasing agents – nonaqueous pentaerythrityl tetrapelargonate; 2,2-bis[[(1- tetraester of pentaerythritol and binders; skin-conditioning agents – oxononyl)oxy]methyl]-1,3-propanediyl pelargonic acid that conforms to the occlusive; viscosity increasing agents – nonanoate; nonanoic acid, 2,2-bis[[(1- structure in figure 1 nonaqueous oxononyl)oxy]methyl]-1,3-propanediyl ester; CAS No. 14450-05-6; trade name: pelemol PTP

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Table 2. Chemical and Physical Properties Property Value Reference Pelargonic Acid Form Oily, colorless liquid Lewis117

Molecular weight 158.24 STN17 Density 0.9055 @ 20 /4 C Lewis117 0.9839 g/cm3 @ 20 C STN17 0.921 ± 0.06 g/cm3 @ 20 C (calculated value) Solubility Miscible with water and methanol Committee of Revision of the United States Pharmacopeial Convention118

Very slightly soluble in water Lewis117 Refractive index 1.4456 @ 20 C and 589.3 nm STN17 Vapor pressure 8.67E-03 Torr (calculated) ACD Labs119 Melting point 12 C Lewis117 12.3 C STN17 Freezing point 12.24 C Lewis117 Boiling point 254.4 C @ 760 Torr ACD Labs119 254.9 ± 3 C @ 760 Torr (calculated) Flash point 100.0 ± 0 C (calculated) ″ Enthalpy of vaporization 52.03 ± 3.0 kJ/mol (calculated) ″ pKa 4.78 ± 0.10 (calculated) ″ logP 3.434 ± 0.184 @ 25 C (calculated) ″ Butylene Glycol Diisononanoate Molecular weight 370.57 ChemDraw120 Melting point 391.86ºK (118.71°C (calculated) " Boiling point 818.52ºK (545.37°C) (calculated) " logP 6.37 (calculated) " Cetearyl Isononanoate (Tegosoft CI) Form Yellowish liquid Evonik Industries121 Refractive index 1.4450 to 1.4500 (specification) Evonik Industries122 Density 0.854 to 0.858 g/ml (specification); Evonik Industries121, 122 ≈ 0.85 g/cm3 @ 68°F 40

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Table 2. Chemical and Physical Properties Property Value Reference Solubility Insoluble in water Evonik Industries121 Viscosity, according to Höppler ≈ 16 @ 25°C Evonik Industries123 (mPas) Volatility 0% in water Evonik Industries 121 Melting temperature < 59°F (15°C) Evonik Industries121 Flash point >212°F (100°C) Evonik Industries121 Solidifications point ≤ 15.0°C (specification) Evonik Industries122 Hydroxyl value ≤ 1.0 mg KOH/g (specification) ″ Iodine value ≤ 1.0 g l/100g (specification) ″ Acid value ≤ 0.2 mg KOH/g (specification) ″ Saponification value 140.0 to 146.0 mg KOH/g ″ (specification) Heavy metals content 20 ppm maximum (Cu; Pb; Sn; Pt; Evonik Industries26 Pd; Hg; As; Cd; Ni); < 1 ppm (Hg; As; Cd; Ni respective) Cellobiose Octanonanoate Molecular weight 1464.08 STN17 Density 1.05 ± 0.1 g/cm3 @ 20 C (calculated) ″

Vapor pressure 0 Torr @ 25 C (calculated) ″ Boiling point 1115.1 ± 65 C @ 760 Torr ″ (calculated) Flash point 371.9 ± 34.3 C (calculated) ″ Enthalpy of vaporization 164.52 ± 3.0 kJ/mol @ 760 Torr ″ (calculated) log P 32.470 ± 0.860 @ 25 C (calculated ) ″ Cetyl Isononanoate Molecular weight 382.66 ChemDraw120 Melting point 398.34ºK (125.19°C) (calculated) ″ Boiling point 829.38ºK (556.23°C) (calculated) ″ logP 9.28 (calculated) ChemDraw120 Cholesteryl Nonanoate Molecular weight 526.88 (calculated) STN17 41

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Table 2. Chemical and Physical Properties Property Value Reference

Density 0.97 ± 0.1 g/cm3 @ 20 C and 760 ″ Torr (calculated) Optical rotation - 30 @ concentration of 1.0 g/100 ml ″ in chloroform Vapor pressure 2.77E-13 Torr @ 25 C (calculated) ″ Melting point 80°C (calculated) ″ Boiling point 576.3 ± 29 C @ 760 Torr ″ (calculated) Flash point 300.8 ± 11.8 C (calculated) ″ Enthalpy of vaporization 86.30 ± 3.0 kJ/mol @ 760 Torr ″ (calculated) logP 14.4245 ± 0.299 @ 25 C (calculated) ″ Diethylene Glycol Diethylhexanoate/Diisononanoate Molecular weight 576.85 ChemDraw120 Diethylene Glycol Diisononanoate Molecular weight 386.57 ChemDraw120 Melting point 414.09ºK (140.94°C) (calculated) ″ Boiling point 890.94ºK (617.79°C) (calculated) ″ logP 5.65 (calculated) ″ Dihydrocholesteryl Nonanoate Molecular weight 528.89 ChemDraw120 Dipentaerythrityl Pentaisononanoate Molecular weight 922.38 ChemDraw120 Ethylhexyl Isononanoate Molecular weight 270.45 ChemDraw120 Melting point 293.18ºK (20.03°C) (calculated) ″ Boiling point 645.9ºK (372.75°C) (calculated) ″ logP 5.91 (calculated) ChemDraw120 Isodecyl Isononanoate Molecular weight 298.50 ChemDraw120 Refractive index 1.437 to 1.439 @ 25 C Nikitakis and McEwen124 (specification) 42

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Table 2. Chemical and Physical Properties Property Value Reference

Specific gravity 0.852 to 0.858 @ 25 /25 C ″ (specification) Acid value 1.0 maximum (specification) ″ Saponification value 175 to 192 (specification) ″ Iodine value 0.5 maximum (specification) ″ Melting point 315.72ºK (42.57°C) (calculated) ChemDraw120 Boiling point 691.66ºK (418.51°C) (calculated) ″ logP 6.68 (calculated) ″ Isononyl Isononanoate Molecular weight 284.48 ChemDraw120 Refractive index 1.4340 to 1.4360 @ 25 C Nikitakis and McEwen124 (specification) Specific gravity 0.849 to 0.855 @ 25 /25 C ″ (specification) Acid value 1.0 maximum (specification) ″ Saponification value 192 to 202 (specification) ″ Iodine value 0.5 maximum (specification) ″ Melting point 304.45ºK (31.3°C) ChemDraw120 Boiling point 668.78ºK (395.63°C) ″ logP 6.27 (calculated) ″ Isononyl Isononanoate (Tegosoft INI) Form Colorless to slightly yellow liquid Evonik Industries125 Density 0.865 g/cm3 at 71.60°F (22°C) ″ Solubility Insoluble in water ″ Viscosity, dynamic 5.5 mPa.s @ 20°C ″ Boiling temperature 273.20 to 280.40°F (138°C) @ 1.3 Evonik Industries125 hPa Flash point 305.60°F (152°C) ″ Hydroxyl value ≤ 5.0 mg KOH/g (specification) Evonik Industries126 Iodine value ≤ 0.50 g l/100 g (specification) ″ Acid value ≤ 0.20 mg KOH/g (specification) ″

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Table 2. Chemical and Physical Properties Property Value Reference Saponification value 185 to 200 mg KOH/g ″ (specification) Water content ≤ 0.30 % (specification) ″ Heavy metals content 20 ppm maximum (Cu; Pb; Sn; Pt; Evonik Industries27 Pd; Hg; As; Cd; Ni); < 1 ppm (Hg; As; Cd; Ni respective) Isostearyl Isononanoate Molecular weight 410.72 ChemDraw120 Melting point 405.88ºK (132.73°C) (calculated) ″ Boiling point 874.7ºK (601.55°C) (calculated) ″ logP 10.02 (calculated) ″ Isotridecyl Isononanoate Molecular weight 340.58 ChemDraw120 Refractive index 1.433 to 1.445 @ 25 C Nikitakis and McEwen124 (specification) Specific gravity 0.859 to 0.861 @ 25 /25 C ″ (specification) Acid value 1.0 maximum (specification) ″ Saponification value 155 to 165 (specification) ″ Iodine value 0.5 maximum (specification) ″ Melting point 349.53ºK (76.38°C) (calculated) ChemDraw120 Boiling point 760.3ºK (487.15°C) (calculated) ″ logP 7.94 (calculated) ″ Neopentyl Glycol Diisononanoate Molecular weight 384.59 ChemDraw120 Melting point 405.55ºK (132.4°C) (calculated) ChemDraw120 Boiling point 838.17ºK (565.02°C) (calculated) ChemDraw120 logP 7.03 (calculated) ″ Pentaerythrityl Tetraisononanoate Molecular weight 697.04 ChemDraw120 Propylene Glycol Diisononanoate Molecular weight 356.54 ChemDraw120

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Table 2. Chemical and Physical Properties Property Value Reference Melting point 365.59ºK (92.44°C) (calculated) ″ Boiling point 795.2ºK (522.05°C) (calculated) ″ logP 6.13 (calculated) ″ Tridecyl Isononanoate Molecular weight 340.58 ChemDraw120 Melting point 364.53ºK (91.38°C) (calculated) ″ Boiling point 760.74ºK (487.59°C) (calculated) ″ logP 8.02 (calculated) ″ Ethylhexyl Pelargonate Molecular weight 270.45 STN 17 Density 0.864 ± 0.06 g/cm3 @ 20°C ″ Mass intrinsic solubility 0.0022 g/l @ 25°C (calculated) ″ Molar intrinsic solubility 0.0000081 mol/l @ 25°C ″ (calculated) Boiling Point 311.8 ± 10.0°C @ 760 Torr ″ (calculated) Melting point 308.18ºK (35.03°C) (calculated) ChemDraw120 Vapor pressure 5.49E-04 Torr @ 25°C (calculated) STN 17 Enthalpy of vaporization 55.28 ± 3.0 kJ/mol @ 760 Torr ″ (calculated) Flash point 144.1 ± 8.8°C (calculated) ″ logP 7.432 ± 0.212 @ 25°C (calculated) ″

Ethyl Pelargonate Molecular weight 186.29 STN17 Density 0.872 ± 0.06 g/cm3 @ 20°C ″ (calculated) Refractive index 1.43367 @ 15°C ″ Mass intrinsic solubility 0.17 g/l @ 25°C (calculated) ″ Molar intrinsic solubility 0.0092 mol/l @ 25°C (calculated) ″ Boiling point 225.5 to 227.5°C; 220.0 ± 0°C @ ″ 760 Torr (calculated)

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Table 2. Chemical and Physical Properties Property Value Reference Melting point -36.7°C (calculated) ″ Vapor pressure 1.16E-01 Torr @ 25°C (calculated) ″ Enthalpy of vaporization 45.64 ± 3.0 kJ/mol @ 760 Torr ″ (calculated) Flash point 94.4 ± 0°C (calculated) ″ logP 4.428 ± 0.206 @ 25°C (calculated) ″ Isobutyl Pelargonate Molecular weight 214.34 (calculated) STN17 Density 0.867 ± 0.06 g/cm3 @ 20°C ″ (calculated) Mass intrinsic solubility 0.041 g/l @ 25°C (calculated) ″ Molar intrinsic solubility 0.00019 mol/l @ 25°C (calculated) ″ Boiling point 248.9 ± 8°C @ 760 Torr ″ (calculated) Melting point 263.1ºK (-10.05°C) (calculated) ChemDraw120 Vapor pressure 2.36E-02 Torr @ 25°C (calculated) STN17 Enthalpy of vaporization 48.61 ± 3.0 kJ/mol @ 760 Torr ″ (calculated) Flash point 104.7 ± 8.3°C (calculated) ″ logP 5.307 ± 0.212 @ 25°C (calculated) ″ Methyl Pelargonate Molecular weight 172.26 (calculated) STN 17 Density 0.8655 g/ cm3 @ 25°C; 0.874 ± 0.06 STN 17 g/cm3 @ 20°C and 760 Torr (calculated) Refractive index 1.4205 @ 20°C and 589.3 nm; ″ 1.41395 @ 25°C and 589.3 nm Mass intrinsic solubility 0.36 g/l @ 25°C (calculated) ″ Molar intrinsic solubility 0.0021 mol/l @ 25°C (calculated) ″ Boiling point 213.5 and 122.0°C; 210.3 ± 3°C @ ″ 760 Torr (calculated) Melting point 244.29ºK (-28.86°C) (calculated) ChemDraw120 Vapor pressure 1.93E-01 Torr @ 25°C (calculated) STN17

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Table 2. Chemical and Physical Properties Property Value Reference Enthalpy of vaporization 44.66 ± 3.0 kJ/mol @ 760 Torr ″ (calculated) Flash point 84.4 ± 0°C (calculated) ″ logP 3.896 ± 0.205 @ 25°C (calculated) ″ Neopentyl Glycol Dicaprylate/Dipelargonate/Dicaprate Molecular weight 1017.55 ChemDraw120 PEG-2 Diisononanoate Molecular weight 574.75 ChemDraw120 PEG-5 Isononanoate Molecular weight 199.35 ChemDraw120 Pentaerythrityl Tetrapelargonate Molecular weight 697.04 (calculated) STN17 Density 0.969 ± 0.06 g/cm3 @ 20°C and 760 ″ Torr (calculated) Mass intrinsic solubility 0.00000077 g/l @ 25°C (calculated) ″ Molar intrinsic solubility 0.0000000011 mol/l @ 25°C ″ (calculated) Boiling point 699.1 ± 50°C @ 760 Torr ″ (calculated) Melting point 12°C (calculated) ″ Vapor pressure 2.16E-19 Torr @ 25°C (calculated) ″ Enthalpy of vaporization 102.35 ± 3.0 kJ/mol @ 760 Torr STN17 (calculated) Flash point 273.5 ± 30.2 °C (calculated) ″ logP 14.879 ± 0.360 @ 25°C (calculated) ″ Isononanoic Acid Molecular weight 158.24 STN17 Density 0.919 ± 0.06 g/cm3 @ 20°C and 760 ″ Torr (calculated) Refractive index 1.4304 @ @ 21°C and 589.3 nm ″ Mass intrinsic solubility 0.52 g/l @ 25°C (calculated) ″ Molar intrinsic solubility 0.0033 mol/l @ 25°C (calculated) ″

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Table 2. Chemical and Physical Properties Property Value Reference Boiling point 248°C @ 765 Torr and 253.4 ± 8°C ″ @ 760 Torr (calculated) Melting point 3 to 5ºC (calculated) ″ Vapor pressure 5.70E-03 Torr @ 25°C (calculated) ″ Enthalpy of vaporization 54.04 ± 6.0 kJ/mol @ 760 Torr ″ (calculated) Flash point 129.7 ± 6.9°C (calculated) ″ logP 3.250 ± 0.194 @ 25ºC (calculated) ″ Isononyl Alcohol Molecular weight 144.25 ChemDraw120 Boiling point 100°C @ 13 Torr STN17 Melting point 64 to 65ºC ″ logP 3.22 SRC127 Isotridecyl Alcohol Molecular weight 200.36 ChemDraw120 Melting point 281.59ºK (8.44°C) (calculated) ″ Boiling point 588.78ºK (315.63°C) (calculated) ″ logP 5.19 (calculated) SRC127 Neopentyl glycol Molecular weight 104.149 STN128 Density 1.1 g/cm3 National Institute for Occupational Safety and Health (NIOSH)129 Impurities Neopentyl glycol formic acid ester Organisation for Economic Coop- and neopentyl glycol isolactic acid eration and Development (OECD)28 ester Water solubility 190 g/100 ml @ 25ºC (65%) ″ Melting point 127 ºC (solvent = benzene) STN128 “ Boiling point 206 ºC @ 747 Torr ″ Flash point 107ºC NIOSH 2010129 Autoignition temperature 388 ºC ″ Explosive limits 1.1 to 11.4 vol% in air ″

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Table 2. Chemical and Physical Properties Property Value Reference Vapor pressure 0.00217522 to 0.0551305 Torr @ ″ 10.74 to 38.14ºC 30 mmHg @140ºC and 760 mmHg OECD 28 @ 211ºC

Enthalpy of fusion 4590 J/mol ″ Enthalpy of vaporization 87,320 J/mol @ 25.24ºC ″ Enthalpy of phase transition 14,100 J/mol ″ Heat capacity 193.48 to 202.21 J/mol *K @ 31.85 ″ to 39.35ºC Thermal decomposition Occurs at > 120ºC in strong base. ″ Thermal decomposition products Methanol, isobutanol, isobutyl ″ aldehyde, formaldehyde, etc. logP 0.12 @25ºC ″

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Table 3. Current Cosmetic Product Uses and Concentrations of Nonanoates and Pelargonic Acid30,31

Product Category 2009 uses (total number of 2009 concentrations (FDA 2009) products in category; FDA 2009) (Personal Care Products Council 2009) (%) butylene glycol diisononanoate Skin care products Skin cleansing creams, lotions, liquids, and pads - 17 Total uses/ranges for butylene glycol diisononanoate - 17 cetearyl isononanoate Eye makeup Eyeliner 2 (684) - Eye shadow 5 (1196) 0.05 Eye lotion 4 (177) - Eye makeup remover 1 (131) - Other 3 (288) - Fragrance products Other 3 (399) 27 to 50 Noncoloring hair products 1 (1097) Tonics, dressings, etc. - Makeup Blushers 1(539) 8 Face powders 2 (613) 0.05 to 11 Foundations 4 (635) 10 Lipstick 1 (1912) - Makeup bases 1 (164) - Other 3 (406 12 Nail care products Creams and lotions 1 (17) - Other 1 (124) - Personal hygiene products Other 2 (514) - Shaving products Aftershave lotion 1 (395) 3 to 6 Preshave lotions 1 (27) - Skin care products Skin cleansing creams, lotions, liquids, and pads 10 (1368) 2 to 3 Face and neck lotions 12 (1195) 0.07 to 8 Body and hand lotions 14 (1513) 3 to 5 Foot powders and sprays 1 (48) - Moisturizers 29 (2039) 2 Night creams and lotions 6 (343) 2 Paste masks (mud packs) 1 (418) - 50

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Table 3. Current Cosmetic Product Uses and Concentrations of Nonanoates and Pelargonic Acid – cont’d

Product Category 2009 uses (total number of 2009 concentrations (FDA 2009) products in category; FDA 2009) (Personal Care Products Council 2009) (%) Other 9 (1244) - Suntan products Indoor tanning preparations 3 (200) - Other 1 (62) - Total uses/ranges for cetearyl isononanoate 123 0.05 to 50 cetearyl nonanoate Skin care products Body and hand lotions - 3 Total uses/ranges for cetearyl nonanoate - 3 cetyl isononanoate Eye makeup Eye lotion - 1 Noncoloring hair products Tonics, dressings, etc. - 1 Skin care products Body and hand lotions - 3 Moisturizers - 5 Other - 1 Total uses/ranges for cetyl isononanoate - 1 to 5 cholesteryl nonanoate Eye makeup Eye lotion 2 (117) - Other 1 (288) - Noncoloring hair products Conditioners - 0.01 Shampoos - 0.01 Makeup Lipstick - 20 to 30 Skin care products Face and neck lotions - 0.04 Moisturizers - 0.06 Paste masks (mud packs) - 0.03 Hair coloring products - Dyes and colors 13 (2481) - Makeup Lipstick 1 (1912) - Makeup bases 1 (164) -

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Table 3. Current Cosmetic Product Uses and Concentrations of Nonanoates and Pelargonic Acid – cont’d

Product Category 2009 uses (total number of 2009 concentrations (FDA 2009) products in category; FDA 2009) (Personal Care Products Council 2009) (%) Skin care products Face and neck lotions 8 (1195) - Moisturizers 3 (2039) - Paste masks (mud packs) 6 (418) - Skin fresheners 1 (285) - Other 2 (1244) - Total uses/ranges for cholesteryl nonanoate 38 0.01 to 30 diethylene glycol diethylhexanoate/diisononanoate Fragrance products Powders 5 (278) 19 Makeup Foundations 1 (635) 18 Lipstick 9 (1912) - Other 1 (406) - Total uses/ranges for diethylene glycol 16 18 to 19 diethylhexanoate/diisononanoate dipentaerythrityl pentaisononanoate Makeup Lipstick - 9 Nail care products Nail polish and enamel - 13 Total uses/ranges for dipentaerythrityl - 9 to 13 pentaisononanoate ethylhexyl isononanoate Eye makeup Eye shadow 3 (1196) 7 to 65 Eye lotion 2 (177) 0.8 Eye makeup remover 1 (131) - Other 2 (288) 12 Fragrance products Other 3 (399) 2 to 5 Noncoloring hair products Conditioners - 1 Rinses - 0.8 Tonics, dressings, etc. 3 (1097) - Other 1 (716) 8 Makeup Blushers 1 (539) 7 Face powders - 3 52

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Table 3. Current Cosmetic Product Uses and Concentrations of Nonanoates and Pelargonic Acid – cont’d

Product Category 2009 uses (total number of 2009 concentrations (FDA 2009) products in category; FDA 2009) (Personal Care Products Council 2009) (%) Foundations 2 (635) 3 Lipstick 9 (1912) - Fixatives 1 (38) - Other 2 (406) 31 (in a face highlighter) Personal hygiene products Other 1 (514) - Shaving products Aftershave lotion 2 (395) 1 Skin care products Skin cleansing creams, lotions, liquids, and pads 1 (1368) 0.8 to 2 Face and neck lotions 5 (1195) 0.04 to 6 Body and hand lotions 31 (1513) 0.04 to 74 Body and hand sprays - 18 Moisturizers 14 (2039) 2 Night creams and lotions 5 (343) 0.02 Other 3 (1244) 0.08 Suntan products Suntan gels, creams, and liquids 1 (156) 7 Indoor tanning preparations 22 (200) 0.07 to 1 Other 1 (62) 0.03 Total uses/ranges for ethylhexyl isononanoate 116 0.02 to 74 isodecyl isononanoate Eye makeup Eye shadow 2 (1196) 21 Eye lotion - 6 Eye makeup remover - 10 Noncoloring hair care products Conditioners - 2 Other 1 (406) - Makeup Blushers 6 (539) 22 to 26 Foundations 1 (635) 59 Lipstick - 0.05 to 18 Rouges - 13 Skin care products Face and neck lotions 5 (1195) - Moisturizers 4 (2039) - Night creams and lotions 1 (343) - Paste masks (mud packs) 2 (418) 2 53

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Table 3. Current Cosmetic Product Uses and Concentrations of Nonanoates and Pelargonic Acid – cont’d

Product Category 2009 uses (total number of 2009 concentrations (FDA 2009) products in category; FDA 2009) (Personal Care Products Council 2009) (%) Other 2 (1244) - Suntan products Suntan gels, creams, and liquids 1 (156) - Other 1 (62) 5 Total uses/ranges for isodecyl isononanoate 26 0.05 to 59 isononyl isononanoate Bath products Oils, tablets, and salts - 15 Soaps and detergents - 8 to 10 Eye makeup Eyebrow pencil - 2 Eye shadow 24(1196) 2 to 18 Eye lotion 8 (177) 3 to 26 Eye makeup remover 2 (131) - Other 13 (288) 6 to 12 (12% in a concealer) Fragrance products Perfumes 4 (569) 26 to 42 Other 2 (399) 21 to 46 Noncoloring hair care products Other 1 (716) - Conditioners - 0.08 Sprays/aerosol fixatives - 0.4 Rinses - 0.03 to 1 Tonics, dressings, etc. - 7 Hair coloring products Bleaches - 33 Makeup Blushers 23 (539) 4 to 17 Face powders 12 (613) 2 to 15 Foundations 28 (635) 3 to 27 Leg and body paints 3 (29) 4 to 57 Lipstick 28 (1912) 8 to 50 Makeup bases 3 (164) 3 to 7 Rouges 3 (99) 12 Makeup fixatives 3 (38) - Other 12 (406) 4 to 6 (4% in a concealer) Nail care products Nail extenders - 0.4 Nail polish and enamel removers - 5 54

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Table 3. Current Cosmetic Product Uses and Concentrations of Nonanoates and Pelargonic Acid – cont’d

Product Category 2009 uses (total number of 2009 concentrations (FDA 2009) products in category; FDA 2009) (Personal Care Products Council 2009) (%) Personal hygiene products Deodorants (underarm) 1 (540) 3 Other 1 (514) - Shaving products Aftershave lotion 2 (395) 3 to 4 Preshave lotions - 22 Skin care products Skin cleansing creams, lotions, liquids, and pads 11 (1368) 0.04 to 21 Face and neck lotions 39 (1195) 0.05 to 17 Face and neck sprays - 6 Body and hand lotions 29 (1513) 5 to 50 Foot powders and sprays - 3 Moisturizers 48 (2039) 3 to 13 Night creams and lotions 10 (343) 2 to 11 Paste masks (mud packs) 2 (418) 2 to 64 Skin fresheners 1 (285) - Other 16 (1244) 1 to 21 Suntan products Suntan gels, creams, and liquids 2 (156) 2 to 9 Indoor tanning preparations 8 (200) 0.3 to 3 Other 4 (62) 0.08 to 21 Total uses/ranges for isononyl isononanoate 343 0.03 to 64 isotridecyl isononanoate Eye makeup Eye shadow - 0.7 Noncoloring hair products Conditioners - 3 Makeup Blushers 15 (539) 4 Face powders 6 (613) 10 Foundations 10 (635) 0.8 to 9 Lipstick 19 (1912) 10 Makeup bases - 5 to 7 Rouges - 4 Other 3 (406) 5 Skin care products Face and neck lotions 2 (1195) - Body and hand lotions 1 Moisturizers 4 (2039) 11 55

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Table 3. Current Cosmetic Product Uses and Concentrations of Nonanoates and Pelargonic Acid – cont’d

Product Category 2009 uses (total number of 2009 concentrations (FDA 2009) products in category; FDA 2009) (Personal Care Products Council 2009) (%) Night creams and lotions 2 (343) - Other 1 (1244) 5 to 51 Suntan products - Suntan gels, creams, and liquids - 0.8 Total uses/ranges for isotridecyl isononanoate 62 0.7 to 51 neopentyl glycol diisononanoate Skin care products Skin cleansing creams, lotions, liquids, and pads - 1 Total uses/ranges for neopentyl glycol diisononanoate - 1 PEG-2 diisononanoate Nail care products Creams and lotions - 2 Total uses/ranges for PEG-2 diisononanoate - 2 PEG-5 isononanoate Skin care products Skin cleansing creams, lotions, liquids, and pads - 1 Total uses/ranges for PEG-5 isononanoate - 1 pentaerythrityl tetraisononanoate Eye makeup Eye lotion - 2 Skin care products Body and hand lotions - 1 Total uses/ranges for pentaerythrityl - 1 to 2 tetraisononanoate polyglyceryl-20 octaisononanoate Skin care products Skin cleansing creams, lotions, liquids, and pads - 3 Total uses/ranges for polyglyceryl-20 - 3 octaisononanoate tridecyl isononanoate Makeup Foundations 1 (635) 9 Total uses/ranges for tridecyl isononanoate 1 9 ethylhexyl pelargonate Hair coloring products Dyes and colors - 5 Eye makeup Eye shadow - 2

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Table 3. Current Cosmetic Product Uses and Concentrations of Nonanoates and Pelargonic Acid – cont’d

Product Category 2009 uses (total number of 2009 concentrations (FDA 2009) products in category; FDA 2009) (Personal Care Products Council 2009) (%) Makeup Blushers 1 (539) - Foundations 1 (635) 25 Skin care products Skin cleansing creams, lotions, liquids, and pads 1 (1368) 2 Body and hand lotions - 4 Total uses/ranges for ethylhexyl pelargonate 3 2 to 25 pentaerythrityl tetrapelargonate Noncoloring hair care products Tonics, dressings etc. - 2 Total uses/ranges for pentaerythrityl tetrapelargonate - 2

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Table 4. Skin Irritation and Sensitization Studies on Pelargonic Acid, Nonanoate Esters, and Related Chemicals

Test Substance Subjects Tested Test Protocol Results References Predictive Tests – Skin Irritation Pelargonic Acid 0.01 M, 0.1 M, 20 male subjects 24 h Al-test® patch Erythematous reactions: 7 Stillman et al.86 0.5 M and 1.0 M (ages not stated): 0.5 application to subjects (0.5 M), 10 (in propanol) M application (10 interscapular area, 10 subjects (1.0 M), and 5 subjects) and 1.0 M applications subjects (0.01 and 0.1 M) (10 subjects); 0.01 M total/dose (0.04 ml and 0.1 M volume; application intradermal injections area [cm2] not (5 subjects) stated); 10 intradermal injections (0.1 ml/injection) total /dose 5%, 10%, 20%, 116 male subjects (21 48 h patch (Al-test® Skin irritation in > 90% Wahlberg and and 39.9% (in to 55 years old) disc) application to of subjects at 48 h and 96 Maibach87 propanol) upper back; 0.04 h post-application (20% ml/dose (application and 39.9% area [cm2] not stated) concentrations); skin irritation in 54.3% of subjects at 48 h and in 48.5% of subjects at 96 h (10% concentration); skin irritation in 12.9% of subjects at 48 h and in 13.9% of subjects at 96 h (5% concentration) 20% (in 16 subjects (10 24 h closed patch (12 Erythema and slight Agner and propanol; pH of males, 6 females; mm-diameter Finn infiltration at 24 h; Serup88 4.3) 29.5 years) chamber) application occasional slight crusting to anterolateral at 48 h surface of both upper arms; dose volume/cm2 not stated 40% , 60%, 70%, 42 male subjects 48 h patch (8 mm- Positive reactions: 2 of 12 Willis et al.89 and 80% (in (mean age of 34 diameter Finn subjects (40% propanol) years): 40% (12 chamber) application concentration); 20 of 32 subjects), 60% (32 to volar forearm (30 subjects (60%); 2 of 32 subjects), 70% (32 µl/cm2) subjects (70%); and 28 of subjects), and 80% 28 subjects (80%) (28 subjects) 20% (in 16 subjects (ages not 24 h closed patch (12 Mean irritation score of 2 Agner and propanol) stated) mm-diameter Finn (moderate positive Serup90 chamber) application reaction) at 24 h; mean to anterolateral score of 1 (weak positive surface of both upper reaction) at 96 h arms; dose volume/cm2 not stated

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Table 4. Skin Irritation and Sensitization Studies on Pelargonic Acid, Nonanoate Esters, and Related Chemicals

Test Substance Subjects Tested Test Protocol Results References Pelargonic Acid 80% (in propan- 10 male subjects 48 h patch (8 mm- Mild to moderate skin Willis et al.130 1-(w/w) 100% (mean age of 35 diameter Finn irritation at 1 h post- propylene years) chamber) application removal glycol) to volar forearm (30 µl/cm2) 20% (in 20 subjects (12 males 24 h closed patch (12 Skin irritation in all Agner and propanol) , 8 females; mean age mm-diameter Finn subjects at 24 h. Serup92 of 27.5 years) chamber) application to flexor side of both upper arms (0.06 ml/dose; application area [cm2] not stated) 20% (in 17 subjects (10 24 h closed patch (12 2 subjects with reactions Agner and propanol) males, 7 females; mm-diameter Finn ranging from score of 0.5 Serup131,131 mean age of 27 years) chamber) application to 2 during winter; 1 to each arm. Seasonal subject with reactions variation (summer ranging from score of 1 to and winter) in skin 2 during summer. No response to significant seasonal pelargonic acid variation in skin response evaluated; dose volume/cm2 not stated 80% (in 10 male subjects 48 h patch (8 mm- Most of the irritation Willis et al.132 propanol, 100% (ages not stated) diameter Finn reactions were mild to propylene chamber) application moderate at 1 h post- glycol) to volar forearm (30 removal µl/cm2) 10%,20%, 40%, 152 female subjects 47 h patch (8 mm- Erythema was more Reiche et al.95 and 80% (in (> 18 years old; 37 diameter Finn severe at 48 h than at 96 h propanol) were atopic chamber) application at sites on the left and to right and left lower right lower back (p < back 0.001). Except for the 20% concentration (on left lower back),decreased erythema with time was noted at each concentration

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Table 4. Skin Irritation and Sensitization Studies on Pelargonic Acid, Nonanoate Esters, and Related Chemicals

Test Substance Subjects Tested Test Protocol Results References 10%, 20%, and 12 subjects (7 males, Cumulative skin Maximum visual irritation Clemmensen et 30% (in 5 females; mean age irritation test score of 12 reported for 2 al.96 propanol) of 25.3) (ROAT). subjects tested with 30% Applications (10 concentration. Pelargonic min) made to volar acid considered quite forearm daily for 5 + irritating at all 4 days (no concentrations tested applications on weekend); 800 µl pipette through 20 mm-diameter circular hole in template Pelargonic Acid 30%, 40%, and 12 subjects (8 Cumulative skin All subjects received Clemmesen et 50% (in females, 4 males; irritation test (Wash irritation score of 2 (scale: al.96 propanol) mean age of 27) test). Test areas (3 x 0 to 15) for at least one 5 cm) on volar test concentration. forearm washed (using oral hygiene sponge) with respective concentrations (dose volume = 4 ml) twice daily for 12 days. 10% (in 12 subjects (8 males, 24 h occlusive patch Irritation reactions (scale: Suihko and isopropanol) 4 females; 18 to 64 (10 mm-diameter 0 to 4+): 2+ (moderate, 5 Serup97 years old) Finn chamber) subjects) and 1+ (mild, 7 application (60 µl) to subjects ) right and left forearm; application area [cm2] not stated Cetearyl Isononanoate 20% Cetiol ® 21 subjects 24 h occlusive No skin irritation Kleber and SN application (Finn Hoffmann- chambers) Dörr64 Cetearyl Nonanoate Undiluted 52 volunteers (males 48 h occlusive patch No skin irritation Symrise chemical and females) application (0.2 g) to GmbH & Co. upper back; KG45 application area [cm2] not stated Neopentyl Glycol Diisononanoate Undiluted 52 volunteers (males Preceding patch test No skin irritation Symrise chemical and females) procedure; GmbH & Co. application area KG 2010c50 [cm2] not stated

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Table 4. Skin Irritation and Sensitization Studies on Pelargonic Acid, Nonanoate Esters, and Related Chemicals

Test Substance Subjects Tested Test Protocol Results References PEG-5 Isononanoate Experimental 53 volunteers 48 h occlusive patch No skin irritation Symrise formulation application (0.2 g) to GmbH & Co. containing 14.5% upper back; KG51 PEG-5 application area isononanoate [cm2] not stated Ethyl Pelargonate Concentration --- 5 min to 5 h applica- No skin irritation Opdyke 37 not stated tion period 12% in --- 48 h closed patch test No skin irritation Opdyke 37 petrolatum) 20% (w/w) in 10 healthy volunteers 24 h occlusive patch No skin irritation Smith et al.98 petrolatum (4 males, 6 females; test 39 to 70 years old) Provocative Tests – Skin Irritation Pelargonic Acid 5%, 10%, 20%, 75 allergic contact 48 h patch (Al-test ® Skin irritation in 93.9% of Wahlberg and and 100% (in dermatitis patients disc) application 33 women tested and Maibach133 propanol) (males and females; (0.04 ml) to upper 93.3% of 48 men tested 20 to 70 years old) back; application area with 20% concentration [cm2] not stated (subjects comprise largest test group). Additional results in table 5 1%, 5%, 10%, 100 hospitalized 48 h patch (Al-test ® Irritation reactions (lowest Wahlberg et 20%, and 39.9% patients with skin disc, saturated with test concentration al.134 (in propanol) disease (54 males, 46 solution) application producing a reaction): females; ages not to upper back; dose 1.0% pelargonic acid (1 stated) volume/cm2 not male; 2 females), 5.0% stated (23 males; 9 females), 10.0 % (18 males; 22 females), 20.0% (12 males; 13 females), and 39.9% (all patients) Skin Irritation and Sensitization Cetearyl Nonanoate Undiluted 106 volunteers (males RIPT. 24 h occlusive No skin irritation or Symrise chemical and females) patch applications, sensitization GmbH & Co. 0.2 g per patch, to KG45 upper back (induction and challenge); application area [cm2] not stated

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Table 4. Skin Irritation and Sensitization Studies on Pelargonic Acid, Nonanoate Esters, and Related Chemicals

Test Substance Subjects Tested Test Protocol Results References Ethylhexyl Isononanoate Undiluted 52 participants (males RIPT same as in Skin irritation in 10 Symrise chemical and females) preceding study, subjects. No allergic GmbH & Co. except semi- contact sensitization KG51 occlusive patches used Isodecyl Isononanoate Makeup product 101 normal subjects RIPT. 24 h 6 subjects with ± or 1+ Clinical containing (18 to 65 years old) semiocclusive patch reaction during induction. Research 51.35% applications to upper 3 subjects with ± reaction Laboratories, back (induction and after challenge. Makeup Inc.101 challenge); dose vol- product had no dermal ume/cm2 not stated irritation or sensitization potential. Isononyl Isononanoate 53 subjects Lipstick RIPT. 24h semi- No skin reactivity Consumer containing 3.55% occlusive patch Product applications,0.2 g per Testing patch, to back (induc- Company 102 tion and challenge); dose volume/cm2 not stated Lipstick 97 subjects " No skin reactivity Consumer containing 3.13% Product Testing Company 103 Neopentyl Glycol Diisononanoate Undiluted 106 volunteers (males RIPT. 24 h occlusive No skin irritation or Symrise chemical and females) patch applications, sensitization GmbH & Co. 0.2 g per patch, to KG50 upper back (induction and challenge); application area [cm2] not stated Predictive Tests - Skin Sensitization Pelargonic Acid 12% in 25 normal subjects Maximization test No sensitization Opdyke 37 petrolatum Cetearyl Isononanoate Liquid makeup 25 normal subjects Maximization test No contact allergy KGL, Inc.104 remover (18 to 65 years old) containing 1.5%

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Table 4. Skin Irritation and Sensitization Studies on Pelargonic Acid, Nonanoate Esters, and Related Chemicals

Test Substance Subjects Tested Test Protocol Results References Cholesteryl Nonanoate Lipstick 28 normal subjects Maximization test No contact allergy KGL, Inc.105 containing (21 to 57 years old) 20.86% Isotridecyl Isononanoate Facial cream 28 normal subjects Maximization test No contact allergy KGL, Inc.106 containing 4.3% (19 to 63 years old) Isodecyl Isononanoate Day cream 26 normal subjects Maximization test No contact allergy KGL, Inc.107 containing 2.6% (26 to 65 years old) Isononyl Isononanoate Eye shadow 26 normal subjects Maximization test No contact allergy KGL, Inc.108 containing (18 to 65 years old) 24.66% PEG-5 Isononanaote Experimental 53 normal subjects RIPT. Patch No allergic contact Symrise formulation applications, 0.2 g sensitization GmbH & Co. containing 14.5% 51 per occlusive patch, KG PEG-5 to upper back isononanoate (induction and challenge); application area [cm2] not stated Ethyl Pelargonoate 12% in 25 normal subjects Maximization test No sensitization Opdyke37 petrolatum Provocative Test – Skin Sensitization Cetearyl Isononanoate Undiluted Cetiol 20 eczema patients RIPT. 24 h No irritation/sensitization Herzberg109 ® SN applications under Beiersdorf test plaster Isotridecyl Alcohol 5% in petrolatum 229 dermatitis 24 h or 48 h patch No sensitization Geier et al.110 patients (mean age = applications 39.3 years)

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Table 5. Reactions in dermatitis patients patch tested with pelargonic acid99

Females Males Total Concentration (%) N Positive N Positive N Positive (%) (%) (%) 5 25 16.0 13 46.2 38 26.3 10 16 56.3 6 83.3 22 63.6 20 33 93.9 15 93.3 48 93.8 100 6 100 6 100 12 100

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Figure 1. Structures/Formulas of Nonanoates/Pelargonates

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CH3 O

H3C CH (CH2)5 C (OCH2CH2)5OH PEG-5 Isononanoate

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78. Hazleton Washington Inc. Mutagenicity test on n-pelargonic acid. In vivo mouse micronucleus assay. MRID# 43603702. Unpublished data (cd-rom) submitted by EPA in response to an FOIA request dated August 31, 2009. 1993;

79. National Toxicology Program (NTP).Testing status: pelagonic acid M070024. Genetic toxicology - Salmonella (A341272) completed.9-7-2009.

80. Symrise GmbH & Co.KG. Summary of human safety studies of Dragoxat 89 (ethylhexyl isononanoate). Unpublished data submitted by the Personal Care Products Council, February 15, 2010. 2010;1-4.

81. CIT. Bacterial reverse mutation test on isononyl isononanoate. Laboratory study no. 29659MMO. Unpublished data submitted by the Personal Care Products Council, May 11, 2010. 2005;

82. Kusakabe, H, Yamakage, K, Wakuri, S, Sasaki, K, Nakagawa, Y, Watanabe, M, Hayashi, M, Sofuni, T., Ono, H., and and Tanaka, N. Relevance of chemical structure and cytotoxicity to the induction of

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chromosome aberrations based on the testing results of 98 high production volume industrial chemicals. Mutation Research. 2002;517:187-198.

83. Renner, H. W. The anticlastogenic potential of fatty acid methyl esters. Mutat Res. 1986;172:265-269.

84. University of Cincinnati Medical Center's Kettering Laboratory. Initial submission: Chronic mouse dermal toxicity study of a mixture of valeric acid, heptanoic acid, pelargonic acid, and tripropylene glycol dicaprylate w/cover letter dated 102792. NTIS Report No.OTS0571597. 1985;

85. Nishikawa, Y. Okabe M. Yoshimoto K. Kurono G. and Fukuoka F. Chemical and biochemical studies on carbohydrate esters. II. Antitumor activity of saturated fatty acids and their ester derivatives against Ehrlich ascites carcinoma. Chem Pharm Bull. 1976;24:387-393.

86. Stillman, M. A. Maibach H. I. and Shalita A. R. Relative irritancy of free fatty acids of different chain length. Contact Dermatitis. 1975;1:65-69.

87. Wahlberg, J. E. and Maibach, H. I. Nonanoic acid irritation: A positive control at routine patch testing? Contact Dermatitis. 1980;6:(2):128-130.

88. Agner, T. and Serup, J. Skin reactions to irritants assessed by polysulfide rubber replica. Contact Dermatitis. 1987;17:(4):205-211.

89. Willis, C. M., Stephens, J. M., and Wilkinson, J. D. Experimentally-induced irritant contact dermatitis. Determination of optimum irritant concentrations. Contact Dermatitis. 1988;18:(1):20-24.

90. Agner, T. and Serup, J. Contact thermography for assessment of skin damage due to experimental irritants. Acta Derm.Venereol. 1988;68:(3):192-195.

91. Willis, C. M., Stephens, C. J., and Wilkinson, J. D. Epidermal damage induced by irritants in man: light and electron microscopic study. J.Invest.Dermatol. 1989;93:(Nov %1989 P695-699):REF

92. Agner, T. and Serup, J. Skin reactions to irritants assessed by non-invasive bioengineering methods. Contact Dermatitis. 1989;%20:(5):352-359.

93. Agner, T. and Serup, J. Seasonal variation of skin resistance to irritants. Br.J.Dermatol. 1989;121:(3):323- 328.

94. Willis, C. M., Stephens, C. Jm, and Wilkinson, J. D. Differential effects of structurally unrelated chemical irritants on the density and morphology of epidermal CD1 positive cells. J.Invest Dermatol. 1990;95:(6):711-716.

95. Reiche, L., Willis, C., Wilkinson, J., Shaw, S., and de, Lacharriere O. Clinical morphology of sodium lauryl sulfate (SLS) and nonanoic acid (NAA) irritant patch test reactions at 48 h and 96 h in 152 subjects. Contact Dermatitis. 1998;39:(5):240-243.

96. Clemmensen, A., Andersen, F., Petersen, T. K., Kalden, H., Melgaard, A., and Andersen, K. E. The irritant potential of n-propanol (nonanoic acid vehicle) in cumulative skin irritation: a validation study of two different human in vivo test models. Skin Res.Technol. 2008;14:(3):277-286.

97. Suihko, C. and Serup, J. Fluorescence confocal laser scanning microscopy for in vivo imaging of epidermal reactions to two experimental irritants. Skin Res.Technol. 2008;14:(4):498-503.

98. Smith, J. S., Macina, O. T, Sussman, N. B., Karol, M. H., and and Maibach, H. I. Experimental validation of a structure-activity relationship model of skin irritation by esters. Quant.Struct-Act.Relat. 2000;19:467-474.

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99. Wahlberg, J. E. and Maibach, H. I. Nonanoic acid irritation - a positive control at routine patch testing? Contact Dermatitis. 1980;6:(2):128-130.

100. Wahlberg, J. E., Wrangsjo, K., and Hietasalo, A. Skin irritancy from nonanoic acid. Contact Dermatitis. 1985;13:(4):266-269.

101. Clinical Research laboratories Inc. Repeated insult patch test of a makeup containing 51.35% isodecyl isononanoate. Unpublished data submitted by the Personal Care Products Council, December 17, 2009. 2009;1-13.

102. Consumer Product Testing Company. Summary of an HRIPT of a lipstick containing 3.552% isononyl isononanoate. Unpublished data submitted by the Personal Care Products Council, November 30, 2009. 2007;

103. Consumer Product Testing Company. Summary of an HRIPT of a lipstick containing 3.128% isononyl isononanoate. Unpublished data submitted by the Personal Care Products Council, November 30, 2009. 2007;

104. KGL Inc. An evaluation of the contact-sensitization potential of a topical coded product in human skin by means of the maximization assay (product contained 1.5% cetearyl isononanoate). KGL protocol 6793. Unpublished data submitted by the Personal Care Products Council, December 17, 2009. 2009;1-10.

105. KGL Inc. An evaluation of the contact-sensitization potential of a topical coded product in human skin by means of the maximization assay (product containied 20.86% cholesteryl nonanoate). KGL protocol 5543. Unpublished data submitted by the Personal Care Products Council, December 17, 2009. 2004;1-10.

106. KGL Inc. An evaluation of the contact-sensitization potential of a topical coded product in human skin by means of the maximization assay (product contained 4.3% isotridecyl isononanoate). KGL protocol 5696. Unpublished data submitted by the Personal Care Products Council, December 17, 2009. 2004;1-11.

107. KGL Inc. An evaluation of the contact-sensitization potential of a topical coded product in human skin by means of the maximization assay (product contained 2.6% isodecyl isononanoate). KGL protocol 6705. Unpublished submitted by the Personal Care Products Council, December 17, 2009. 2009;1- 10.

108. KGL Inc. An evaluatioin of the contact-sensitization potential of a topical coded product in human skin by means of the maximization assay (product contained 24.66% isononyl isononanoate). KGL protocol 6793. Unpublished data submitted by the Personal Care Products Council, December 17, 2009. 2008;1-10.

109. Herzberg, J. J. Repeated insult patch test of five substances, including Cetiol SN (cetearyl isononanoate) (English translation). Unpublished data submitted by the Personal Care Products Council, June 1, 2010. 1967;

110. Geier, J, Lessmann, H., Frosch, P. J., Pirker, C, Koch, P., Aschoff, R., Richter, G., Becker, D., Eckert, C., Uter, W., Schnuch, A., and and Fuchs, T. Patch testing with components of water-bsased metalworking fluids. Contact Dermatitis. 2003;49:85-90.

111. Willis, C. M., Stephens, C. Jm, and Wilkinson, J. D. Differential effects of structurally unrelated chemical irritants on the density of proliferating keratinocytes in 48 h patch test reactions. J.Invest Dermatol. 1992;99:(4):449-453.

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112. Forsey, R. J., Shahidullah, H., Sands, C., McVittie, E., Aldridge, R. D., Hunter, J. A., and Howie, S. E. Epidermal Langerhans cell apoptosis is induced in vivo by nonanoic acid but not by sodium lauryl sulphate. Br.J.Dermatol. 1998;139:(3):453-461.

113. Lindberg, M., Farm, G., and Scheynius, A. Differential effects of sodium lauryl sulphate and non-anoic acid on the expression of CD1a and ICAM-1 in human epidermis. Acta Derm.Venereol. 1991;71:(5):384-388.

114. Mathias, C. Gt, Maibach, H. I., and Conant, M. A. Perioral leukoderma simulating vitiligo from use of a toothpaste containing cinnamic aldehyde. Arch.Dermatol. 1980;116:(10):1172-1173.

115. Le Coz, C. J. and Bressieux, A. Allergic contact dermatitis from cetearyl isononanoate. Contact Dermatitis. 2003;48:(6):343-343.

116. Goossens, A., Verbruggen, K., Cattaert, N., and Boey, L. New cosmetic allergens: isononyl isononanoate and trioleyl phosphate. Contact Dermatitis. 2008;59:(5):320-321.

117. Lewis, R. J. Dangerous Properties of Industrial Materials. 2000;10th:(3):2739New York: Wiley- Interscience.

118. Committeee of Revision of the United States Pharmacopeial Convention. The United States Pharmacopeia. 2003;26th:2498Rockville, MD: United States Pharmacopoeial Convention, Inc.

119. Advanced Chemistry Development (ACD) Labs.ACD Labs calculated values. In: Scienific and Technical Information Network's Registry file.2010.

120. ChemDraw [computer program]. 2002;Cambridge, MA: Cambridge Soft Corporation.

121. Evonik Industries. Material safety data sheet on Tegosoft CI (cetearyl isononanoate). Unpublished data submitted by the Personal Care Products Council, October 23, 2009. 2007;1-8.

122. Evonik Industries. Product specification on Tegosoft CI (cetearyl isononanoate). Unpublished data submitted by the Personal Care Products Council, October 23, 2009. 2008;1-1.

123. Evonik Industries. Further product information on Tegosoft CI (cetearyl isononanoate). Unpublished data submitted by the Personal Care Poducts Council, October 23, 2009. 1999;1-3.

124. Nikitakis, J. M. and McEwen, G. N. Jr. CTFA Compendium of Cosmetic Ingredient Composition - Specifications. 1990;Washington, D.C.: CTFA.

125. Evonik Industries. Material safety data sheet on Tegosoft INI (isononyl isononanoate). Unpublished data submitted by the Personal Care Products Council, October 23, 2009. 2009;1-8.

126. Evonik Industries. Product specification on Tegosoft INI (isononyl isononanoate). Unpublished data submitted by the Personal Care Products Council, October 23, 2009. 2008;1-1.

127. SRC.InteractiveLogKow (KowWin) Demo.2010.

128. Scientific and Technical Information Network (STN.Beilstein file.2010.

129. National Institute for Occupational Safety and Health (NIOSH). International Chemical Safety Cards. Neopentyl glycol. 2010;1-3.

130. Willis, C. M., Stephens, C. J. M., and Wilkinson, J. D. Epidermal Damage Induced by Irritants in Man: A Light and Electron Microscopic Study. Journal of Investigative.Dermatology. 1989;93:(5):

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131. Agner, T. and Serup, J. Seasonal variation of skin resistance to irritants. Br.J.Dermatol. 1989;121:(3):323- 328.

132. Willis, C. M., Stephens, C. J., and Wilkinson, J. D. Differential effects of structurally unrelated chemical irritants on the density and morphology of epidermal CD1+ cells. J.Invest Dermatol. 1990;95:(6):711-716.

133. Wahlberg, J. E., Wrangsjo, K., and Hietasalo, A. Skin irritancy from nonanoic-acid. Contact Dermatitis. 1985;13:(4):266-269.

134. Wahlberg, J. E., Wrangsjo, K., and Hietasalo, A. Skin Irritancy From Nonanoic Acid. Contact Dermatitis. 1985;13:(4):Hospitalized-acid.

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Final Report

Stearyl Heptanoate and Related Stearyl Alkanoates as Used in Cosmetics

August 31, 2010

The 2010 Cosmetic Ingredient Review Expert Panel members are: Chairman, Wilma F. Bergfeld, M.D., F.A.C.P.; Donald V. Belsito, M.D.; Curtis D. Klaassen, Ph.D.; Daniel C. Liebler, Ph.D.; Ronald A Hill, Ph.D. James G. Marks, Jr., M.D.; Ronald C. Shank, Ph.D.; Thomas J. Slaga, Ph.D.; and Paul W. Snyder, D.V.M., Ph.D. The CIR Director is F. Alan Andersen, Ph.D. This report was prepared Monice Fiume, Senior Scientific Analyst/Writer.

Supplement Panel Book 1 Page 288 Alkyl Esters Supplement Book 1 TABLE OF CONTENTS

Abstract ...... 1 Introduction ...... 1 Chemistry ...... 1 Definition and Structure ...... 1 Use ...... 1 Cosmetic ...... 1 Non-Cosmetic ...... 2 General Biology ...... 2 Absorption, Distribution, Metabolism, and Excretion ...... 2 Animal Toxicology ...... 2 Acute Toxicity ...... 2 Short-Term Toxicity ...... 2 Subchronic Toxicity ...... 2 Dermal Irritation ...... 2 Sensitization ...... 3 Phototoxicity ...... 3 Comedogenicity ...... 3 Ocular Irritation ...... 3 Genotoxicity ...... 3 Carcinogenicity ...... 3 Tumor Promotion ...... 3 Clinical Assessment of Safety ...... 3 Irritation and Sensitization ...... 3 Phototoxicity ...... 4 Comedogenicity ...... 4 Summary ...... 4 Discussion ...... 4 Conclusion ...... 5 Figure 1. Figure map of stearyl alkanoates...... 6 Table 1. Definitions, synonyms, and cosmetic functions...... 7 Table 2. Frequency of use according to duration and exposure...... 8 References ...... 9

Supplement Panel Book 1 Page 289 Alkyl Esters Supplement Book 1 ABSTRACT Stearyl heptanoate is a stearyl alkanoate (the ester of stearyl alcohol and heptanoic acid) that functions in cosmetics as a skin conditioning agent. Stearyl caprylate, stearyl palmitate, stearyl stearate, stearyl behenate, and stearyl olivate are also stearyl alkanoates with similar structures and functions. The safety information included in this review applies to stearyl heptanoate or some of the components of the alkanoate esters, and the available information supports the safety of all of the related stearyl alkanoates. No toxicological concerns were raised by any of this information. The Expert Panel concluded that stearyl heptanoate, stearyl caprylate, stearyl palmitate, stearyl stearate, stearyl behenate, and stearyl olivate are safe in the present practices of use and concentration.

INTRODUCTION Stearyl heptanoate has previously been reviewed by the Cosmetic Ingredient Review (CIR) Expert Panel. In 1995, the safety assessment was published with the conclusion that stearyl heptanoate is “safe for use as a cosmetic ingredient in the present practice of use.”1 A search of the currently available published literature found no new studies. Although there were no new data available to prompt a rereview of stearyl heptanoate, the Expert Panel did determine that the safety data included in the 1995 assessment of stearyl heptanoate are applicable to the safety of stearyl caprylate, stearyl palmitate, stearyl stearate, stearyl behenate, and stearyl olivate. For that reason only, the safety assessment of stearyl heptanoate was reopened to include these ingredients. Brief summaries of information from the original safety assessment are included for the absorption, toxicology and clinical sections. Since limited data were available for the original assessment on stearyl heptanoate, and because stearyl alkanoates are metabolized by esterases in the body to stearyl alcohol and the related fatty acid,2-4 relevant data from the report on stearyl alcohol were included in the 1995 review to support safety. As such, brief summaries of relevant information from the stearyl alcohol report5 are also included. (It was concluded that stearyl alcohol was safe as used in cosmetics. Stearyl alcohol was used at concentrations ≤50%.) A CIR safety assessment is available for palmitic acid and stearic acids,6 the fatty acid components of stearyl palmitate and stearyl stearate, respectively. Summaries from the assessment on these components are therefore included in this report. (It was concluded that palmitic and stearic acid were safe as used in cosmetics. Palmitic acid was used at concentrations of ≤25% and stearic acid was used at concentrations >50%.)

CHEMISTRY Definition and Structure The definition, synonyms, and function in cosmetics for stearyl heptanoate, stearyl caprylate, stearyl palmitate, stearyl stearate, stearyl behenate,7 and stearyl olivate8 are given it Table 1. The structure map of these chemicals is depicted in Figure 1.

USE Cosmetic All of the stearyl alkanoates included in this report function as skin conditioning agents, with some having additional uses.7 (See Table 1.) According to information supplied to the Food and Drug Administration (FDA) as part of the Voluntary Cosmetic Registration Program (VCRP), at the time of the original assessment, stearyl heptanoate was used in 121 cosmetic formulations in 1994.1 Concentrations of use were not reported to the FDA at the time of the original safety assessment, but unpublished data reported use at up to 25%. Since then, frequency as well as concentration of use has decreased. Recent VCRP data indicate that stearyl heptanoate is used in 102 cosmetic formulations,9 and results of a survey of current use concentrations conducted by the Personal Care Products Council (Council) indicate that stearyl heptanoate is now used at concentrations of 0.07-8%.10 Current and historical use data are given in Table 2. It was reported to the VCRP that stearyl caprylate and stearyl stearate are used in 20 and 22 cosmetic formulations, respectively, and stearyl olivate is used in 1 cosmetic formulation.9 According to the concentration of use survey conducted by the Council, stearyl caprylate is used at concentrations of 0.1-1% and stearyl stearate is used at concentrations of 0.02- 4%.11 No uses were reported for stearyl olivate in response to the survey. Stearyl palmitate and stearyl behenate are not reported within VCRP data.9 However, according to the survey of use concentrations conducted by the Council, stearyl palmitate and stearyl behenate are used in formulations applied to the area around the eyes at concentration of 3% and 0.02%, respectively.11 (See Table 2 for use details).

Supplement Panel Book 1 Page 290 Alkyl Esters Supplement Book 1 Stearyl stearate is used in products that may be inhaled, and effects on the lungs that may be induced by inhaled products containing these ingredients are of concern. The aerosol properties that determine deposition in the respiratory system are particle size and density. The parameter most closely associated with deposition is the aerodynamic diameter, da, defined as the diameter of a sphere of unit density possessing the same terminal settling velocity as the particle in question.

In humans, particles with an aerodynamic diameter of ≤ 10µm are respirable. Particles with a da from 0.1 - 10µm settle in the 12,13 upper respiratory tract and particles with a da < 0.1 µm settle in the lower respiratory tract. Particle diameters of 60-80 µm and ≥80 µm have been reported for anhydrous hair sprays and pump hairsprays, respectively.14 In practice, aerosols should have at least 99% of their particle diameters in the 10 – 110 µm range and the mean particle diameter in a typical aerosol spray has been reported as ~38 µm.15 Therefore, most aerosol particles are deposited in the nasopharyngeal region and are not respirable. Stearyl heptanoate, stearyl caprylate, stearyl palmitate, stearyl stearate, stearyl behenate and stearyl olivate are listed in the European Union inventory.16 Non-Cosmetic Stearyl palmitate and stearyl stearate are included on the list of indirect additives used in food contact substances.17

GENERAL BIOLOGY Absorption, Distribution, Metabolism, and Excretion Stearyl alcohol is found naturally in various mammalian tissues and is readily converted to stearic acid.5 Stearyl alcohol is poorly absorbed from the gastrointestinal tract. Fatty acids are absorbed, digested, and transported in animals and humans.6 Palmitic and stearic acids are primarily transported via the lymphatic system. In digestion studies, stearic acid was the most poorly absorbed of the common fatty acids. From the Final Report on the Safety Assessment of Stearyl Alcohol, Oleyl Alcohol, and Octyl Dodecanol.5 From the Final Report on the Safety Assessment of Oleic Acid, Lauric Acid, Palmitic Acid, Myristic Acid, and Stearic Acid.6

ANIMAL TOXICOLOGY

Acute Toxicity 1 The oral LD50 of stearyl heptanoate, concentration not specified, was >16 ml/kg for albino Sprague-Dawley rats. A cosmetic formulation containing 0.7% stearyl heptanoate, given orally to Sprague Dawley rats, had an LD50 >5.0 g/kg. 5 The LD50 of stearyl alcohol was >8 g/kg when given orally to Holtzman albino rats. The oral LD50 of palmitic acid was >10 g/kg for rats, and 5 g/kg of a shaving cream containing 2.2% palmitic acid given orally and dermally was non- 6 toxic. The oral LD50s of cosmetic formulations containing ≤13% stearic acid were >5g/kg for rats. Mild erythema and slight induration of skin was observed in an acute study with rabbits and guinea pigs dosed with ≤100 mM stearic acid. From The Final Report on the Safety Assessment of Stearyl Heptanoate.1 From the Final Report on the Safety Assessment of Stearyl Alcohol, Oleyl Alcohol, and Octyl Dodecanol.5 From the Final Report on the Safety Assessment of Oleic Acid, Lauric Acid, Palmitic Acid, Myristic Acid, and Stearic Acid.6 Short-Term Toxicity No toxic effects were observed when chicks were fed diets containing 50% stearic acid for 4 wks.6 Application of 18 mmol% palmitic acid to the external ear canal of rabbits for 6 wks caused slight irritation for the first 2 wks. In a 4-wk study, application of a formulation containing 20% stearic acid to abraded and intact skin of rabbits produced slight edema and desquamation, but it did not raise any toxicologic concerns. From the Final Report on the Safety Assessment of Oleic Acid, Lauric Acid, Palmitic Acid, Myristic Acid, and Stearic Acid.6 Subchronic Toxicity No evidence of systemic toxicity was observed with a formulation containing 8.0% stearyl alcohol in a 3-mos percutaneous study using rabbits.5 In 13-wk dermal toxicity studies, application of formulations containing ≤5% stearic acid to rabbits or rats produced minimal to mild irritant effects.6 From the Final Report on the Safety Assessment of Oleic Acid, Lauric Acid, Palmitic Acid, Myristic Acid, and Stearic Acid.6 Dermal Irritation Undiluted stearyl heptanoate was mildly irritating to rabbit skin.1 Undiluted stearyl alcohol applied under occlusion was minimally to mildly irritating to rabbit skin.5 Palmitic acid, as supplied and in formulation at ≤74%, and stearic acid, as supplied and in formulation at ≤65%, were at most mildly irritating.6 From The Final Report on the Safety Assessment of Stearyl Heptanoate.1 From the Final Report on the Safety Assessment of Stearyl Alcohol, Oleyl Alcohol, and Octyl Dodecanol.5 From the Final Report on the Safety Assessment of Oleic Acid, Lauric Acid, Palmitic Acid, Myristic Acid, and Stearic Acid.6 2

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Sensitization A cosmetic formulation containing 1.5% stearyl heptanoate was not a sensitizer using guinea pigs.1 A deodorant formulation containing 24.0% stearyl alcohol was not a contact sensitizer using guinea pigs.5 A formulation containing 1% stearic acid was a weak, grade 1, sensitizer using guinea pigs, while in a maximization study, a cosmetic formulation containing 3.5% stearic acid was not a sensitizer using guinea pigs.6 From The Final Report on the Safety Assessment of Stearyl Heptanoate.1 From the Final Report on the Safety Assessment of Stearyl Alcohol, Oleyl Alcohol, and Octyl Dodecanol.5 From the Final Report on the Safety Assessment of Oleic Acid, Lauric Acid, Palmitic Acid, Myristic Acid, and Stearic Acid.6 Phototoxicity Two cosmetic formulations containing 2.8% stearic acid were not phototoxic or photoallergenic using guinea pigs.6 From the Final Report on the Safety Assessment of Oleic Acid, Lauric Acid, Palmitic Acid, Myristic Acid, and Stearic Acid.6 Comedogenicity A cosmetic formulation containing 1.5% stearyl heptanoate produced slight to moderate comedogenicity in rabbit ears.1 Stearyl alcohol, concentration not known, was not comedogenic when applied to the ear canal of rabbits.5 From The Final Report on the Safety Assessment of Stearyl Heptanoate.1 From the Final Report on the Safety Assessment of Stearyl Alcohol, Oleyl Alcohol, and Octyl Dodecanol.5 Ocular Irritation Undiluted stearyl heptanoate was classified as a category 3 (not defined) eye irritant using rabbits.1 Cosmetic formulations containing 0.7 or 1.5% stearyl heptanoate were not primary eye irritants. Undiluted stearyl alcohol was minimally irritating to rabbit eyes.5 Palmitic acid, as supplied and in formulations at ≤19.4%, produced at most mild irritation, but were generally non-irritating to rabbit eyes.6 Stearic acid, as supplied and in formulation at ≤65%, produced some initial mild ocular irritation. From The Final Report on the Safety Assessment of Stearyl Heptanoate.1 From the Final Report on the Safety Assessment of Stearyl Alcohol, Oleyl Alcohol, and Octyl Dodecanol.5 From the Final Report on the Safety Assessment of Oleic Acid, Lauric Acid, Palmitic Acid, Myristic Acid, and Stearic Acid.6

GENOTOXICITY A mixture containing >50% stearyl heptanoate, at ≤5000 µg/plate, did not induce base pair or frameshift mutations with or without metabolic activation.1 The mixture, ≤5000 mg/kg, did not have a clastogenic effect in an in vivo micronucleus test using mice. Stearyl alcohol, concentration tested not given, was not mutagenic in an Ames assay with or without metabolic activation.5 Stearic acid, 50 µg/plate, was not mutagenic in an Ames assay, and ≤500 µg/ml did not induce mitotic aneuploidy. From The Final Report on the Safety Assessment of Stearyl Heptanoate.1 From the Final Report on the Safety Assessment of Stearyl Alcohol, Oleyl Alcohol, and Octyl Dodecanol.5

Carcinogenicity Stearic and palmitic acid, 50 mg/kg/day given as a dietary supplement to rats (duration not given), did not produce carcinogenic effects.6 From the Final Report on the Safety Assessment of Oleic Acid, Lauric Acid, Palmitic Acid, Myristic Acid, and Stearic Acid.6

Tumor Promotion Stearyl alcohol did not promote tumor formation in female Swiss mice when initiated with 7,12-dimethylbenz[a]- anthracene.5 From the Final Report on the Safety Assessment of Stearyl Alcohol, Oleyl Alcohol, and Octyl Dodecanol.5

CLINICAL ASSESSMENT OF SAFETY Irritation and Sensitization Stearyl heptanoate, tested at an unspecified concentration and in formulations containing ≤1.5%, was not an irritant or sensitizer, and daily patch testing of formulations containing 8-24% stearyl heptanoate was essentially non- to slightly irritating.1 Undiluted stearyl alcohol was a mild irritant in a single insult occlusive patch test.5 Four of 1664 subjects had an allergic reaction to 30% stearyl alcohol in liquid paraffin. Cosmetic formulations containing 2.2% palmitic acid were non-irritating and non-sensitizing.6 Formulations containing ≤40% stearic acid were generally non-irritating, except for one formulation containing 2.8% stearic acid, which was moderately irritating in a 21-day cumulative irritation assay. In sensitization assays, ≤13% stearic acid in cosmetic formulations produced some irritation at induction, but generally no reaction at challenge. From The Final Report on the Safety Assessment of Stearyl Heptanoate.1 From the Final Report on the Safety Assessment of Stearyl Alcohol, Oleyl Alcohol, and Octyl Dodecanol.5

Supplement Panel Book 1 Page 292 Alkyl Esters Supplement Book 1 From the Final Report on the Safety Assessment of Oleic Acid, Lauric Acid, Palmitic Acid, Myristic Acid, and Stearic Acid.6

Phototoxicity A cosmetic formulation containing 2.2% palmitic acid was non-photosensitizing in a study using 52 subjects and cosmetic formulations containing 1-13% stearic acid were not phototoxic, photosensitizing, or photoallergenic in studies using 20-101 subjects.6 From the Final Report on the Safety Assessment of Oleic Acid, Lauric Acid, Palmitic Acid, Myristic Acid, and Stearic Acid.6

Comedogenicity A cosmetic formulation containing 1.5% stearyl heptanoate was not comedogenic.1 From The Final Report on the Safety Assessment of Stearyl Heptanoate.1

SUMMARY Stearyl heptanoate has previously been reviewed by the Cosmetic Ingredient Review (CIR) Expert Panel, and, in 1995, it was concluded that this ingredient was safe as used. A recent search of the literature published since that report was issued did not find any new data. However, the Panel agreed that stearyl caprylate, stearyl palmitate, stearyl stearate, stearyl behenate, and stearyl olivate were similar to stearyl heptanoate, and that the original safety assessment on stearyl heptanoate could address the safety of these stearyl alkanoates as well. Therefore, a rereview was initiated to include these ingredients. The stearyl alkanoates reviewed in this safety assessment all function as skin conditioning agents; some have a few other functions. In 1994, it was reported to the Food and Drug Administration (FDA) that stearyl heptanoate was used in 121 cosmetic formulations and, according to unpublished data, it was being used at up to 25%. As of 2010, the frequency of use has decreased to 102, and, according to an industry survey, use concentrations have decreased and are now ≤8%. It has been reported to the FDA that stearyl caprylate and stearyl stearate are used in 20 and 22 cosmetic formulations, respectively, and that stearyl olivate is used in 1 cosmetic formulation. According to an industry survey, stearyl caprylate is used at concentrations of 0.1-1%, stearyl stearate is used at 0.02-4%, and stearyl olivate is not reported to be used. Stearyl palmitate and stearyl behenate are not reported to be in use according to VCRP data, however, industry data reports that these ingredients are used in eye products at concentrations of 3 and 0.02% respectively. Since published data were not found for these ingredients, summaries from the original safety assessment on stearyl heptanoate and from previous CIR assessments on some of the components of these stearyl alkanoates, i.e. stearyl alcohol, palmitic acid, and stearic acid, were included in this review. The information from these previous assessments did not raise any toxicological concerns, and there were no concerns of genotoxic effects. Also, these ingredients were at most mildly irritating to the skin, and they did not appear to be sensitizers. Palmitic acid and stearic acid were not phototoxic. Stearyl heptanoate produced slight to moderate comedogenicity in rabbit ears, but was not comedogenic for humans. At most, mild ocular irritation was seen in testing with any of these ingredients.

DISCUSSION A search of the currently available published literature found no new studies on stearyl heptanoate. However, the Expert Panel did determine that the safety data included in the 1995 assessment of stearyl heptanoate are applicable to the safety of stearyl caprylate, stearyl palmitate, stearyl stearate, stearyl behenate, and stearyl olivate. For that reason only, the safety assessment of stearyl heptanoate was reopened to include these ingredients. In the original safety assessment on stearyl heptanoate, the Panel noted that although animal irritation testing was done at 100%, sensitization testing was performed using a maximum concentration of only 1.5% stearyl heptanoate. The Panel concludes that all the available data suggest that stearyl heptanoate would not be a sensitizer. The same would apply to the stearyl alkanoates now included in this rereview. In the original safety assessment, it was also noted that mild ocular reactions were seen in ocular irritation testing with undiluted stearyl heptanoate. The Panel is still of the opinion that stearyl heptanoate, as used in cosmetic formulations, would not produce significant ocular irritation. The same is true for the related stearyl alkanoates. Stearyl stearate is used in cosmetic formulations that can be inhaled. The potential adverse effects of inhaled ingredients depend on the specific chemical species, the concentration and the duration of the exposure and their site of deposition within the respiratory system. In practice, aerosols should have at least 99% of their particle diameters in the 10 – 110 µm range and the mean particle diameter in a typical aerosol spray has been reported as ~38 µm. Particles with an aerodynamic

Supplement Panel Book 1 Page 293 Alkyl Esters Supplement Book 1 diameter of ≤ 10µm are respirable. In the absence of inhalation toxicity data, the panel determined that stearyl stearate can be used safely in products that are inhaled, because the product particle size is not respirable.

CONCLUSION The CIR Expert Panel concluded that stearyl heptanoate, stearyl caprylate, stearyl palmitate, stearyl stearate, stearyl behenate, and stearyl olivate are safe in the present practices of use and concentration.

Supplement Panel Book 1 Page 294 Alkyl Esters Supplement Book 1 Figure 1. Figure map of stearyl alkanoates.

Supplement Panel Book 1 Page 295 Alkyl Esters Supplement Book 1 Table 1. Definitions, synonyms, and cosmetic functions.

INGREDIENT DEFINITION SYNONYMS COSMETIC FUNCTION (CAS No.)

Stearyl Heptanoate the ester of stearyl alcohol and heptanoic acid heptanoic acid, octadecyl ester skin conditioning agent – occlusive (66009-41-4)

Stearyl Caprylate the ester of stearyl alcohol and caprylic acid octanoic acid, octadecyl ester skin conditioning agent – occlusive (18312-31-7)

Stearyl Palmitate the ester of stearyl alcohol and palmitic acid (q.v.) hexadecanoic acid, octadecyl ester binder; emulsion stabilizer; hair conditioning agent; humectant; (2598-99-4) opacifying agent; skin conditioning agent - miscellaneous

Stearyl Stearate the ester of stearyl alcohol and stearic acid octadecanoic acid, octadecyl ester skin conditioning agent – occlusive; viscosity increasing agent - (2778-96-3) nonaqueous Stearyl Behenate the ester of stearyl alcohol (q.v.) and behenic acid (q.v.) docosanoic acid. octadecyl ester skin conditioning agent – occlusive (24271-12-3) Stearyl Olivate the ester of stearyl alcohol and the fatty acids derived from skin conditioning agent; emollient; surfactant – emulsifying agent (no CAS No.) Olea europaea (olive) oil (q.v.); olive oil is composed of a number of fatty acids, primarily oleic, linoleic, palmitic, palmitoleic, and stearic acids

Supplement Panel Book 1 Page 296 Alkyl Esters Supplement Book 1 Table 2. Frequency of use according to duration and exposure. Stearyl Heptanoate Freq. of Freq. Of Use Conc of Use Conc of Use

Use 19941 20109 (%) 19931 (%) 201010 TOTAL 121 102 2-25 0.07-8 Duration of Use Leave-On 115 99 2-25 0.07-8 Rinse Off 6 3 NR 0.7-3 Exposure Type Eye Area 36 17 NR 0.5-8 Possible Ingestion 7 8 5-25 3-8 Inhalation 2 1 NR NR Dermal Contact 117 100 2-25 0.07-8 Deodorant (underarm) 0 NR NR 0.07 Hair - Non-Coloring 4 2 NR NR Hair - Coloring NR NR NR NR Nail NR NR NR NR Mucous Membrane NR 2 NR NR Bath Products NR NR NR NR Baby Products NR NR NR NR

Stearyl Caprylate Stearyl Palmitate Stearyl Stearate Stearyl Behenate Stearyl Olivate Freq. of Conc. of Use Freq. of Conc. of Use Freq. of Use Conc. of Use Freq. of Use Conc. of Use Freq. of Conc. of Use Use 20109 (%) 201011 Use 20109 (%) 201011 20109 (%) 201011 20109 (%) 201011 Use 20109 (%) 201011 TOTAL 20 0.1-1 NR 3 22 0.02-4 NR 0.02 1 NR Duration of Use Leave-On 19 0.3-1 NR 3 20 0.02-4 NR 0.02 NR NR Rinse Off 1 0.1-0.6 NR NR 2 2 NR NR 1 NR Exposure Type Eye Area 2 ≤1 NR 3 5 ≤1 NR 0.02 NR NR Possible Ingestion 2 NR NR NR 5 ≤1 NR NR NR NR Inhalation NR NR NR NR 1 NR NR NR NR NR Dermal Contact 20 ≤1 NR NR 16 ≥4 NR NR 1 NR Deodorant (underarm) NR NR NR NR NR NR NR NR NR NR Hair - Non-Coloring NR NR NR NR 1 3 NR NR NR NR Hair - Coloring NR NR NR NR NR NR NR NR NR NR Nail NR NR NR NR NR NR NR NR NR NR Mucous Membrane 1 NR NR NR 2 2 NR NR NR NR Bath Products NR NR NR NR NR NR NR NR NR NR Baby Products NR NR NR NR NR NR NR NR NR NR

NR - no use reported to the VCRP or by industry for this category 8

Supplement Panel Book 1 Page 297 Alkyl Esters Supplement Book 1 REFERENCES 1. Andersen FA (ed). Final report on the safety assessment of Stearyl Heptanoate. J Am Coll Toxicol. 1995;14(6):498- 510.

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4. Riemenschneider, W. Organic Esters. 6th ed. New York: Wiley-VCH, 2002.

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6. Elder RL. Final report on the safety assessment of oleic acid, lauric acid, palmitic acid, myristic acid, and stearic acid. J Am Coll Toxicol. 1987;6:(3):321-401.

7. Gottschalck, TE and Bailey, JE eds. International Cosmetic Ingredient Dictionary and Handbook. 13th ed. Washington, DC: Personal Care Products Council, 2010.

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10. Personal Care Products Council. Concentration of use of Stearyl Heptanoate. Response to an industry survey. 12-17- 2009.

11. Personal Care Products Council. Concentration of use additions to the Stearyl Heptanoate report. Response to an industry survey. 5-13-2010.

12. James AC, Stahlhofen W, Rudolf G, and et al. Deposition of inhaled particles. Annals of the ICRP. 1994;24:(1-3):231- 232.

13. Oberdörster G, Oberdörster E, and Oberdörster J. An emerging discipline evolving from studies of ultrafine particles. Environ Health Perspect. 2005;113:(7):823-839.

14. Bower D. 1999. Unpublished information on hair spray particle size provided at the September 9 Expert Panel meeting.

15. Johnson MA. The influence of particle size. Spray Technology and Marketing. 2004;November:24-27.

16. European Commission.European Commission Enterprise and Industry. Cosmetics - Cosing. 2010. Accessed 5-4- 2010.

17. Food and Drug Administration.List of indirect additives used in food contact substances. 1-8-2010. http://www.accessdata.fda.gov/scripts/fcn/fcnNavigation.cfm?rpt=iaListing&page=28. Accessed 5-4-2010.

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