Translation 3055

0.tedud;Le.. FISHERIES AND MARINE SERVICE

Translation Series No , 3055

On reductive tendency.of sterol. contents in edible oils. ,and fats, during commercial'processing proéedures

by H. Kanematsu, T. Maruyama, I. Niiya, M. IMamUra, and T. Matsu• oto

— Shokuyo Yushi noS,eisei - KakO:Kotei_ni olceru Suterin.no Ky6do ni tàùite

From: Yukagaku. . (Oil Chemistry 22(10 ) • 814817

Translated,hythe Tranalatien HUreaU(HE/u) • Services Division Mult•lingual • Departmentof the SeOretary of:State of , Cane4s

Department of theSpvironment .Fisherles'and Marine Service - Halifax LabbratorY • . Halifax, N.É.

12 pages tyPescript. DEPARTMENT OF THE SECRETARY OF STATE SECRÉTARIAT D'ÉTAT

TRANSLATION BUREAU BUREAU DES TRADUCTIONS

MULTILINGUAL SERVICES DIVISION DES SERVICES

DIVISION MULTILINGUES

Fe•111 TRANSLATED FRONI TRADUCTION DE INTO EN Japanese English AUTHOR - AUTEUR H. Kanematsu, T. Maruyama, 1. Nilya, M. Imamura, aid T. Matsumoto

TITLE IN ENGLISH - TITRE ANGLAIS On Reductive Tendency of Sterol Contents In Edible Oils and Fats during Commercial Processing Procedures

TITLE IN FOREIGN LANGUAGE (TRANSLITERATE FOREIGN CHARACTERS) Shokuyo Yushi no Seisei Kako Kotei TITRE EN LANGUE ÉTRANGÉRE (TRANSCRIRE EN CARACTÈRES ROMAINS) ni okeru Suterin no Kyodo ni tsuite

REFEREPICE IN FOREIGN LANGUAGE (NAME OF BOOK OR PUBLICATION) IN FULL. TRANSLITERATE FOREIGN CHARACTERS. RÉFERENCE EN LANGUE ÉTRANGÉRE (NOM DU LIVRE OU PUBLICATION), AU COMPLET, TRANSCRIRE EN CARACTÉRES ROMAINS.

YUkagaku

REFERENCE IN ENGLISH - RÉFÉRENCE EN ANGLAIS

Journal of Oil Chemistry

PAGE NUMBERS IN ORIGINAL PUBLISHER - ÉDITEUR DATE OF PUBLICATION NUMÉROS DES PAGES DANS DATE DE PUBLICATION L'ORIGINAL

YEAR ISSUE NO. pp. S14-7 VOLUME NUMÉRO PLACE OF PUBLICATION ANNE NUMBER OF TYPED PAGES LIEU DE PUBLICATION NOMBRE DE PAGES DACTYLOGRAPHIÉES

Tokyo, Japan 1973 22 12 12

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SOS.200.10-6 (REV. 2/68) 7850-21-029-533 3 DEPARTMENT OF THE SECRETARY OF STATE SECRÉTARIAT D'ÉTAT TRANSLATION BUREAU BUREAU DES TRADUCTIONS

MULTILINGUAL SERVICES DIVISION DES SERVICES DIVISION MULTILINGUES f---e-44 CLIENT'S NO. DEPARTMENT DIVISION/BRANCH CITY N0 DU CLIENT MINISTÉRE DIVISION/DIRECTION VILLE

Environment Fisheries Service Ottawa, Ont.

" BUREAU NO. LANGUAGE TRANSLATOR (INITIALS) N° DU BUREAU LANGUE TRADUCTEUR (INITIALES)

JUN - 3 1974 784552 Japanese HE / 23

On Reductive Tendenc of Sterol Contents in Edible Oils and Fats during CommerciarProcessing Procedures

Hiromu Kanematsu*, Takenori Maruyama * , Isao Niiya* , Masao Imamura* and Taro Matsumoto**

LU * Japan institute of Oils & Fats, Other Foods Inspection, t) Foundation (27-8, Nihonbashi Hama-cho, 3-chome, g-L1> E • Chuo-ku, Tokyo) c 0 z 0 0 -0 ** Faculty of Science & Engineering, Nihon University (Surugadai-Kanda, Chiyoda-ku, Tokyo) f- a 0 2- • For English summary, see p. 13. D Lu U.. c 1 Preface

The method employed to determine the rate at which animal and vegetable oils and fats combine in oil and fat materials used for food products such as margarine and shortening,is by estimation from the compositional ratio of animal 1-3 sterol and vegetable sterol. However, due to various reasons, among them the fact that sterol content varies with the species of oil and fat, the compositional ratio of sterols in most cases does not reflect the composition of oil and fat 2 materials.

SOS-200-10-31 .. 2 ..

Many reports have been written on the subject of sterol 4-7 content in animal and vegetable oils and fats , but we believe that the evaluation of these reports as they stand is hazardous for the following reasons among others. First, depending upon the method of measurement used, there is a 7 difference in quantitative value ; and second, the records for the processing of the oils and fats analyzed are not always clear.

As a part of the study on additions and components present in minute quantity in margarine and shortening, the present writers examined the behavior of sterols during the processing procedures of oils and fats, as well as the difference In sterol content in the various species of oil and fat, and hereby report as follows.

2 Samples and Method of Experiment p.815

2.1 Samples

The samples were obtained from refineries of oil and fat. They consisted of oils and fats from four commercially processed species of animal oil and fat, and eight commercially processed species of vegetable oil and fat, in the forms of crude oil) and specimens secured at each stage in the processes of deacidificatioh, decoloration, hydrogenation, and deodoriza-

'tion. The samples were furnished by the refineries of Asahi Denka Kogyo CO. Ltd., Miyoshi Yushi Co. Ltd., Tsukijima Shoktihin Kogyo Co. Ltd., Nihon Yushi. Co. Ltd., Ueda Seiyu Co. Ltd., Taiyo Yushi Co. Ltd., Kano Yushi Co. Ltd. and Nikka Yushi Co. Ltd. The properties of crude oil used for the experiment, and the changes in the rates of iodation, oxidation, and peroxi- * dation caused by refining and hydrogenation are shown by the mean values of the various species in Table 1.

2.2 Method of Experiment We measured the content of unsaponifiable matter by "The Standard Analytical Method for the Examination of Oils and 8 Fats" , and we employed for sterols the method which we pre- viously reported. That is to say, on the understanding that unsaponifiable matter would be dealt with next, we sought the compositional ratio of sterols by performing Thin Layer Chromatography and Gas Liquid Chromatography. Furthermore, we prepared a separate sample by adding a fixed quantity of cholesterol.or vegetable sterol as a control substance, and also performed TLC and GLC on it. We thon calculated the sterol content from the difference in the peak areasof the two, and the known amount of the control substance added. The conditions of TLC and GLC are as follows.

* Translatorls Note: In section 2.1 ("Samples") and section 3.1 ("Sterol Content and the Content of Unsaponifiable Matter in Each Species'of Oil and Fat"), "refining" refers to deacidification, decoloration, - , and deodorization,. and "hydrogenation" to hydrogenation and deodorization as the footnotes of Tables 1 and 2 illustrate. 4

Table-1 Properties and effect of processing on various oils and fats (mean value).

mp (°C) S.V. I.V. A.V. P.O.V. Species n Re- ^ t Hydroge- Re- Hydroge. Re- Hydroge- Crude Crude Crude Crude Crude fined • nated fined^•t natedt" finedt'' nated^et

Beef tallow 39.3 196.8 49.2 47.5 34.7 6.39 0.06 0.13 j 7.55 0.73 0.78 Pork fat 6? 29.8 197.3 63.9 62.3 51.0 5.18 0.08 0.11 7.40 1,59 0.55 186.6 150.9 - 78.6 5.20 - 0.10 5.76 - 0.52 Fish oil t ..4 - .• Whale oil 2 - 187.2 125.2 - !• -72.5 8.89 - 0.10 10.39 , - 0.44 Coconut oil 2 24.1 258.5 8.6 7.9 1.6 6.15 0.07 0.07 1.03 0.57 0.05 Palm kernel oil 1 27.1 246.7 • 18.5 16.8 ; - 11.39 0.07 - 10.06 0.67 - Palm oil 2 36.7 200.1 53.6 51.8 44.9 6.90 0.08 0.15 11.32 0.45 0.64 Soybean oil 1 2 - 196.7 129.4 130.6 70.6 1.21 0.10 0.19 11.15 1.34 0.41 Cottonseed oil 1 - 197.6 113.7 112.1 70.3 2.03 0.10 0.08 3.08 1.26 0.44 Kapok oil 1 - 199.0 102.6 104.3 - 14.12 0.10 - 7.50 1.90 - Rapeseed oil 1 - 177.4 105.9 106.5 - 0.85 0.12 - •. 6.71 0.74 - Rice bran oil 1 - 173.4 96.9 107.2 16.37 0.18 - 10.08 Î 1.95

(a) Deacidified, decolorized and deodorized (b) Hydrogenated and deodorized

Table-2 Comparison of sterol and unsaponifiable matters content in crude oils and fats with those in processed with or without hydrogenationg.

Sterol (rzg/ Uaraponifiable . uaaucr^ \/al Sample Re- Hydroge. Re- Hydroge- Crude Crude finedr•t nated^6t finedt•' nated"'

A 1.90 1.11 - 0.83 0.50 B 2.08 1.04 - 0.75 0.46 - Beef tallow C 2.03 1.18 - 0.98 0.59 - D 1.94 - 0.90 0.84 - 0.56 A 1.39 0.50 - 1.05 0.42 - I B •1.09 0.46 - 0.73 0.43 C. 1.16 0.63 - 1.06 0.47 Pork fat D 1.10 0.54 - 1.05 0.49 E 0.83 0.41 - 0.83 0.46 - F 1.03 - 0.38 0.71 - 0.43 A 4.24 - 1.84 1.91 - 1.04 B 4.95 - 1.86 1.88 1.16 Fish oil C 5.30 - 1.96 2.11 - .1.03 D 5.44 - 1.90 2.19 - I 1.08 A 5.19 - 2.82 1.81 - 1.16 Whale oil { ll B 4.00 2.61 1.68 1.04 f A 1.09 0.94 0.54 0.43 0.36 0.31 Coconut oil t B 1.06 0.88 0.45 0.34 0.31 0.30 Palm kernel oil 1.27 0.86 - 0.47 0.33 f A 0.65 0.32 - 0.93 0.33 Pal m .m oil B 0.98 0.36 1.02 0.60 A 3.69 2.82 1.88 2.54 1.78- 1.24 Soybeaa oil 1 B 3.68 3.01 - 2.98 1.30 - Cottonseed oil 4.58 3.85 2.41 1.30 0.90 0.74 Kapok oil 3.84 2.27 - 1 0.83 0.62 ! Rapeseed oil 6.85 i 5.72 ( - 1.39 1.05 Rice bran oil -_I 21.77 10.55 - 5.79 2.50

(a) Desicidified, decolorized and deo^orized (b) Hydro},enated and deodorizcd TLC: Supporting phase hieselge1 G 0.3mm thick Developing Solvent Chloroform: ethyl ester: acetic acid (94:5:1)

GLC: Column 3mm x lm Glass Column p.816 Filler 2% OV-17/ehromosorb W (DMCS) 80/100 mesh

3 Result of the Expnriment and Conclusions

3.1 Sterol Content and the Content of Unsa,onifiable Matter in Each Species of Oil and Fat Table 2 illustrates the sterol content and the content of unsaponifiable matter in the crude, refined and hydrogenated oils of each of the four species of animal oil and fat and eight species of vegetable oil and fat used in the experiment. When we studied the sterol content in crude oil, we observed a difference even within the sanie species of oil and fat, but the difference was comparatively small. However j that difference was relatively large in different species, and we believed that this largely reflected the special characteristics of each species. Palm oil had the lowest sterol content, followed by coconut oil, pork fat, and palm kernel oil, which showed a value of approximately 1 mg/g, a relatively small content. On the other hand, rice bran oil had the highest content s - being overwhelmingly larger than those of other species of oil and fat, and more than twenty times greater than the aforemen- tioned oils and fat. In addition to rice bran oil, rapeseed oil, fish oil, whale oll, cottonseed oil et al. also had a comparatively high content, showing a value of approximately 4-5 mg/g. Refined and hydrogenated oil3had a lower sterol content than crude oil. The content was especially low after hydrogenation, thereby proving that sterol content is decreased by refining and hydrogenation* With regards to the content of unsaponifiable matter, rice bran oil had a high level, but the other species dis- played no great difference. The decreasing rate of the content of unsaponifiable matter proved almost identical with that of the sterols in the refining and hydrogenation procedures.

3.2 The Compositional Ratio of Sterols of Ve etable Oils and Fats As for sterols in edible oils and fats, in general animal oils and fats contain cholesterol. Many vegetable oils and fats, on the other hand, contain several different types of sterols, the main types being campesterol, stigmasterol and e-sitosterol. Table 3 illustrates the compositional ratio of sterols in the eight species of vegetable oil employed in the present experiment.

Table-3 Sterol composition in 8 species of vegetable oils (5',5).

Sample Choie- Campe- Stigma- ,S-Sito. Brissica sterol sterol sterol sterol sterol ...... ___ Coconut I A 1.1 7.7 15.7 75.5 0 ûii 1. B 1.3 8.4 17.3 73.0 0 Palm kernel oil 3.2 4.9 18.3 73.6 0 { A 6.0 21.1 14.4 58.5 0 Palm oil B 2.2 21,1 12.5 64.2 0 soybean f A 0 19.9 21.7 58.4 0 oil 1 B . 0 19.6 23.0 57.4 0 :::.'ottonseed oil Tr. 13.4 1.3 85.3 0 Kapok oil 0 11.3 Tr. 88.7 0 Rapeseed oil 0.9 30.1 0 56.8 12.2 Ltice bran oil 0 19.9 18.0 62.1 0 7

Stigmasterol detected in rapeseed oil and kapok oil was almost negligible in quantity. Apart from the three types of sterols mentioned above, however, cholesterol was detected in coconut oil, palm kernel oil, palm oil and rapeseed oil, while brassicasterol was detected in rapeseed oil. 2, 9-11 Many reports have been written on the subject of sterol composition in vegetable oils and fats, and cholesterol has been detected in some of these oils and fats. In a previous 2 report we stated that even if cholesterol is detected in an oil mixture of unknown composition, it does not prove that the mixture is a combination of animal oils and fats; and the present experiment confirmed this fact. In the previous report we also explained that since stigmasterol was not detected in cottonseed oil, this was one of the characteristics of cottonseed oil. However, though minute in quantity, stigmasterol was detected in the findings of the 11 present experiment. Itoh et al. have also reported finding traces of stigmasterol.

3.3 Chano;es in the Comoositional Ratio of 3terols Caused by Processino Procedures We examined the manner in which the compositional ratio of sterols in vegetable oils and fats was changed by processing. Since all vegetable oils and fats revealed almost identical tendencies, we took palm oil B processed with hydrogenation, and rapeseed oil processed without hydrogenation as representative

examples. The results are shown in Table 4.

Table-4 Comparison of sterol compositions in vegetable oils and fats at various stages of processing (96).

processing ! Clvale- Campe. I Stigma- P-Sito- Brassies- Sample sterol steroll sterol sterol sterol

Crude 2.0 . 21.8 12.1 64.1 0 Deacidified 2.5 22.2 11.8 63.5 0 Palm oil Decolorized 2.4 20.2 13.6 63.8 0 B Hydrogenated 2.3 20.7 12.6 64.4 6 Deximized 2.0 20.5 12.5 65.0 0

Crude I 1.5 29.6 0 56.4 12.5 Rapeseed Deacidified 0.8 30.8 0 55.4 13.0 oil 1Decolorized j 0.8 29.4 0 57.4 12.4 De•tximized 0.5 30.5 0 58.0 11.0_

••-•

In both cases, no major change occurred in the compositional

ratio of sterols before and after each processing procedure. p,817 Accordingly, we concluded that the decrease in sterol content caused by processing is not connected with the type of sterol

involved. •

3.4 212.Lmes_111._he Sterol Content Caused by Processing Procedures As for the decrease which does occur in the sterol content of oils and fats during refining and hydrogenation (explained

in 3.1), in order to represent the rate of decrease vis-...vis the

sterol content in crude oil, we have in Table 5 outlined the results of the analysis of several representative oils and fats 9

in the order in which each procedure stage takes place.

Table--5 Remaining ratios of sterol content in oils and fats at various stages of processing (°o).

^aci- Decolo- Sample Crude Hs^dro- IDeodo- I difxdl rized genated rized ! Pork fat-D 100.0 87.0 78.3 - 49.2 Palm kernel oil 100.0 92.3 58.4 67.3 Palm oil-A 100.0 E5.9 74.r 49.6 Cottonseed oil 100 . 0 96 . 7 94 . 0 }. - 8-1 . 1 Beef tallo^v-D 100.0 86.4 78.3 53.6 46.2 Fish oil-C 100.0 85.8 69.3 44.1 37.0 Coconut oil-A 100.0 94.9 88.6 G8.3 49.8 Soybean oil-A 100.0 93.4 91.5 52.4 r 50.9

Generally, the sterol content in oils and fats decreased gradually from procedure to procedure through deacidification, decoloration, hydrogenation, and deodorization. A significant decrease was observed, however, during the procedure stages of hydrogenation-and deodorization.

The question of decoloration caused by activated terra 12 alba was investigated by Niewiedomski , who sought the rates at which sterol decreased or vanished because of the presence of a quantity of terra alba. He explained that dissolution also occurred with the adsorption of sterol. It is also generally known that during deodorization, a comparatively large amount of sterol is included in its eluate.

Accordingly, there is adsorption of alkali oil scyhesterol during deacidification, and adsorption and dissolution during decoloration. We further believe that sterol is eliminated from oils and fats in the form of vapor distillation during deodorization. - 10-

It is not clear what causes the decrease in sterol content during hydrogenation, but given the fact that the content of unsaponifiable matter has on the whole not decreased during this procedure, we suppose that sterol was transformed 9 into an unsaponifiable matter other than sterol. Eisner further added hydrogen directly to unsaponifiable matter, and claimed that the side-chain unsaturated bond of stigmasterol was reduced to form B-sitosterol. However, the present experiment was conducted for an industrial purpose, and the hydrogen addition was as oils and fats, while excessive hydrogenation was not performed. As a result, we believe that the addition of selective hydrogen revealing the compositional change of sterol did not occur. We observed that of the different species of oil and fat, animal oils and fats and palm oil tended to have a higher rate of decrease of sterols caused by processing.than coconut oil, palm kernel oil and .liquid vegetable oils. This difference in the rate of decrease is believed to be due to variations in the conditions of the processing procedures (such as in the temperature and length of the deodorizing procedure) for oils of the lauric acid system or liquid vegetable oil, which are often taken in their pure form in order to make the most of their distinctive flavor, and for palm oil or animal oils and fats which have to be highly processed in order to be used as materials for commercially processed food. Given the above, it is clear that sterol content in oils and fats, apart from variations within the same species of oil and fat, show a different value depending upon the records of the processing of the sample oils and fats, as well as upon the conditions of the processing procedures. Accordingly, we believe that it is rather difficult to estimate the composition of oil and fat materials used for food products from the compositional ratio of steroband sterol content except in limited cases, such as the determination of animal oils and

fats in vegetable oils and fats or the compositional ratio of a general outline. •

Finally, we would like to express our gratitude to Mr. Kazuaki Ushigusa and M. Mariko Yanagawa.for cooperating with us in this analysis. A summary of the present report was read at the .Twenty-fifth Science Lecture Meeting of the Japanese Society for Food Hygiene (May 18, 1973, Tokyo).

(Received July 30, 1973.) BIFLIOGRAPHY

1) J. Eisner, J.L. Iverson, A.K. Mozingo, D. Firestone, J^As soc. Of f ic . Agr. Chem.

2) Tmamura, Niiya, Maruyama, Terao, Journal of Food Hygiene, 9, 112_(1968),

3) A. Karleskind, F. Audiau, J. P. 4Valff, Rev. France Corps Gras, 12, 399 (1965); 13, 165 (1966).

4) Tsumura, Keita, Sasaki, Journal of Food H,ygiene, 6, 440 (1965).

5) E. Fedeli, A. Lanzani, P. Capella, G. Jacini; J. Amer. Oil Chemists' Soc., 43, 254 (1966).

6) L.N. Norica, E. Barbara, J. Amer. Oi1. Chemists' Soc., 43, 168 (1966).

7) Kanematsu, Maruyama, Yanagawa, Niiya, Imamura, Journal of Food H,yg3.ene, 13s 286 (1972).

.8) Standard Analytical Method for the Examination of Oils and Fats, ed. Japan Institute of Oil Chemîstry Asakura Shoten, 1966) p.163.

9) J. Eisner, D. Firestone, J. Assoc. Offic, Agr. Chem., 46, 542 (1963).

10) D.F. Johnson, R.D. Bennet, E. Heftman, Science, 140, 198 (1963). ll) T. Itoh, T. Tamura, T. Matsu_'noto, J. Amer. Oil Chemists' Soc., 50, 122 (1973). , 12) H. Nievriadomski, 01eaginïeu.Y, As 175 (1958); Sotoyama, Journal _of Oil ChemistrY, 21, 596 (1972).

13) Ota, Journal of Oil Chemi str.y, 14 835 ( 1970). S RY

In order to examine the behavior of sterols during the processing procedures of oils and fats, four species of animal fat and eight species of vegetable oil were submitted to the processes of deacidification, decoloration, hydrogenation and deodorization. Samples were taken at each of these steps; analyzed for sterols, and results obtained i,vere as follows : 1) Campesterol, stigmasterol and e-sitosterol were detected in vegetable oils and the change of their compositional ratio was nearly negligible during the processing procedures. 2) A minute quantiy of cholesterol was detected in coconut oil, palm kernel oil, palm oil and rapeseed oil. Stigmasterol was not detected in rapeseed oil and kapok oil, and a minute quantity was detected in cottonseed oil. Brassicasterol was petected in rapeseed oil. 3) Sterol content in oils and fats decreased gradually with progress in processing procedures and the decrease was marked especially in hydrogenation and deodorization processes. The rate of this decrease in vegetable oils and oils of lauric acid system was smaller than that in animal fats and Palm oil. 4) Sterol content in purified oils and in purified and hydrogenated oils was characteristic to the species of oils and fats but was also affected by conditions of purification and hydrogenation. In crude oils, sterol content was the smallest in palm oil, being ca. 1mg/g, while the content in rice bran oil was larger, being ca. 20 mg/g.