Novel ingredients - fragrances

MAYUREE KANLAYAVATTANAKUL, NATTAYA LOURITH*, JITPINAN JANWATTANAPOL *Corresponding author School of Cosmetic Science, Mae Fah Luang University, Chiang Rai 57100, Thailand

Nattaya Lourith Extraits and absolutes of Thai Plumeria obtusa L. flowers and their preference for cosmetics

KEYWORDS: Plumeria obtusa; pagoda tree; plumeria; enfleurage; preference.

Comparative volatile constituents and odour preference of Plumeria obtusa was done by enfleurage and Abstract solvent extractions. Two enfleurage bases, one consisting of spermaceti wax and palm oil and the other consisting of white beeswax and sunflower oil, were prepared. The extraits of each base were analyzed using (GC-MS) and compared to the absolutes obtained from maceration in n-hexane and ether. The same aromatic components were found in each extract, which were linalool, nerolidol, benzyl benzoate, trans-farnesol and benzyl alcohol. Enfleurage in spermaceti wax and palm oil was the most preference fragrance (42.6%) with the least difference (38.8%) from the fresh flower as evaluated in 50 Thai volunteers.

Introduction saturated fat (pomade) is removed afterwards. The fat base used is important in enfleurage. It must be odourless with high ragrance applications including perfumes and consistency that offers a semi-hard surface to allow sufficient aromatherapy, are increasing and are presented in a absorption of the fragrances and simple removal of the flower- F large variety of products for personal care, cleansing, retaining fat surface (4). There are a number of complicated, and washing, as well as in consumer goods such as toilet labour-intensive steps involved in the enfleurage process, paper, paper tissues, candles etc. In addition, fragrance despite an advance in the techniques compared to the older, applications for indoor air modification such as in shopping more traditional techniques, and these are in addition to those malls, offices, and restaurants are also becoming important required for synthetic aroma materials. True natural fragrances in current society. Production levels to supply a greater produced by enfleurage, particularly flower oils, are of demand are, therefore, increasing (1). Furthermore, the particular high-quality (4,5). There is evidence that enfleurage number of fragrances in each application product is higher has been in use since the time of the ancient Egyptians, for particular perfumes (2). Fragrance materials are balanced who had not yet discovered the art of distillation, and that by the perfumer with the enfleurage has been used intention to create a cosmetic primarily to prepare perfumed product pleasing the users’ oils. The ancient Egyptians expectations. Of which applied pomade directly to natural odourants are mainly their hair, as is documented (6). in demand particularly floral Today, this type of “pomade” notes (3). would not be considered Enfleurage or cold fat very useful; it would therefore extraction is one of the be further extracted using classical methods for aroma absolute alcohol (ethanol) to preparation from flowers produce an absolute, which in addition to expression, would then be further treated distillation and solvent to remove that solvent, leaving extraction. Enfleurage is just the extrait (5). This process based on the absorption is more common now, with of the volatile oil present in Bulgaria, Egypt, Algeria, Sicily the fragrant flowers into the (Italy), and particularly Grasse fat over a period of time (France) producing highly- (which can vary according quality and expensive natural to the flowers). The fragrant flower oils by this method (4).

H&PC Today - household and Personal Care today, Vol. 8(4) July/August 2013 25 Plumeria spp., commonly called plumeria, pagoda tree, added, separately. The maceration tank was sealed from temple tree, or West Indian jasmine or leelawadee in Thai, is light and air exposure and kept under room temperature similar to other oriental flowers. It has a unique and extremely for 24 h. For three consecutive days, extracted flowers were pleasant odour of floral note. This note is gaining increased replaced with the fresh ones every morning. Afterwards, interest in aromatherapy and spa applications, which is vacuum filtration was done and partitioned with denatured one sector of the perfumery industry (7). The flower is varied alcohol. The removal of the solvent was conducted under in colour, either yellow or pink, and it is large part of Thai reduced pressure at economy, cultivated primarily for decorative gardening. 35 °C to obtain an absolute. Although there are several plumeria hybrids that vary in colour, the main cultivars are white (Plumeria obtusa) and GC-MS Analysis red (Plumeria rubra) flowers. The common name ofP. obtusa An aliquot (5 μL) of each sample (50 mg) in 1 mL of CH2Cl2 originates from the pagoda or temple tree that is widely (Fisher, Loughborough, UK) was analyzed using a gas planted in the Buddhist temple areas. This oriental flower has chromatograph (Agilent 6890N, California, USA) equipped a sweet, nuanced fragrance that elicits a claiming sensation, with an HP-5MS (Agilent 122-5532, 30 m × 250 μm, 0.25 μm and therefore it is challenging to extract the closet resemble film thickness) column and mass spectrophotometer (Agilent natural odour. Therefore, the natural fragrance preparation 5973N). The oven program started at 60 °C and increased to of P. obtusa in this study was conducted by enfleurage and 300 °C at a rate of 7 °C/min. Helium was used as the carrier solvent extraction (maceration). The aroma profiles were gas at a constant flow rate of 1.0 mL/min at a pressure of verified, and the preferences were evaluated in comparison 9.32 psi (64 kPa). The injector was kept at 220 °C and sample with fresh flowers. measurements were made in a split mode (split ratio: 100:1). The compounds were identified by comparison with the reference mass spectra from the MS-Willey7n.1database. Materials and Methods The bases and the solvent were additionally analyzed for background cut-off. Those whose mass quality pattern higher Plant material than 90% were confirm identified (8). Plumeria obtusa flowers cultivated in Chiang Rai, Thailand, were harvested in the morning at the beginning of its Preference Test blossoming stage from April to July. The prepared aromas were evaluated to see if they were similar to the fresh P. obtusa flower by a sensory test (9). Fifty Enfleurage base preparation non-smoking, healthy Thai males and females aged 19 – 24 Spermaceti wax (Namsiang, Bangkok, Thailand) was mixed years old without olfactory disorders or fragrance and pollen with palm oil (Thanakorn, Samutprakarn, Thailand) to prepare allergies were included in this preference test. All recruited an enfleurage base. A base consisting of white beeswax subjects were informed about the study both in writing and (Namsiang) and sunflower oil (Thanakorn) was also prepared. verbally, and each signed a written consent form that was Both bases were formulated in a 3:2 ratio. The warm melt approved by the ethical committee of the Mae Fah Luang bases (250 mL, each) were poured into individual aluminum University prior to enrollment in the study. trays (4.5 × 17.0 cm). Four aroma samples consisting of 2 extraits and 2 absolutes (20 μL) diluted in mineral oil (1:10, v/v) were adsorbed on filter Flower oils preparation paper (Whatman No. 1, New Jersey, USA) that had been cut Enfleurage in a square shape (1 × 1 cm). The filter paper was placed in P. obtusa petals (35 g) were strewn by hand on the top of an amber vial that was wrapped in aluminum foil, separately, the enfleurage base layer, wrapped in aluminum foil and with a sniff port diameter of 1 mm. A sniff test of each sample paraffin film to protect the chassis from light exposure. The (1 min) was done, with a 3 min resting period before the pile of the chassis was kept below ambient temperature next sniff. This preference test was performed in a controlled for 24 h. Defleuraged flowers were replaced daily with fresh environment room, thus eliminating any interfering factors, ones every morning over a period of 14 days. The pomade and it was conducted by a volunteer who was not associated obtained was removed with a spatula and extracted three with any fragrance or scent products. times with denatured alcohol (Merck, Darmstadt, Germany) The Likert scale was used to evaluate odour quality (worst using a separatory funnel. The alcoholic fragrance solution – best = 1 – 5) and difference (least – highest = 1 – 5) in was kept under 20 °C for 1 h before filtration. These processes a comparison with fresh P. obtusa flowers. The score was were repeated until the final filtrate gave a clear solution that summarized, averaged, and calculated into a percentage of was the absolute de pomade. The absolute de pomade was quality and difference (mean/5 × 100) (10). further concentrated under vacuum at 35 °C to produce the extrait.

Maceration Fresh-blooming flowers (15 g) were gently placed in an Erlenmeyer flask (1,000 mL), to which 300 mL of n-hexane (Merck) or ether (Mallinckrodt, Table 1. Appearance, extractive yield and preference of P. obtusa flower extracts Florida, USA) was

26 H&PC Today - household and Personal Care today, Vol. 8(4) July/August 2013 Results and Discussion

Plumeria obtusa is accounted in perfumery applications for the impact of its fine, sweet, and elegant fragrance. The plant was originally cultivated around the areas of Buddhist temple areas to keep them fumigated and also to provide a sense of calm and holistic benefits. Its common name is rooted in these origins. In addition, its fresh flowers have been widely used in traditional Thai aroma materials in several forms, e.g., aroma candles and incense. Furthermore, the extremely pleasant odour of the oriental floral note is in high demand (7). Therefore, aroma profiles of this fragrant flower were studied. The harvesting of P. * RT: Retention Time obtusa flowers for enfleurage sample no. 1: enfleurage from spermaceti wax and palm oil base, sample no. 2: enfleurage from white and solvent extractions were beeswax and sunflower oil base, sample no. 3: absolute from ether, sample no. 4: absolute from n-Hexane carried out in the cooler temperatures of the morning, Table 2. Volatile constituents (%) of P. obtusa aroma extracts when the aroma compounds were present at their highest concentration. The aroma preparations were conducted and other fatty alcohols and fatty acids. The benefits of this from April to July, during the highest flowering session of clean and innocuous wax were that there was less chance the plant. of deterioration as a result of the extraction, providing a clear Spermaceti wax was used in the presenting enfleurage and colourless aroma extract (11). Beeswax was additionally because it is white, translucent, slightly unctuous, and free of chosen for the enfleurage base preparation due to its wide rancidity due to its containing cetyl palmitate, cetyl alcohol, application in aromatherapy products (12), in addition to its

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8 www.icis.com/europeansurfactants or & +44 (0) 20 8652 4659. Media Partners: Please quote promote code when booking your place. H&PC Today - household andUWT54357 Personal Care today, Vol. 8(4) July/August 2013 27 Magazine cosmetic importance based on physicochemical characters and 57.4%), no. 3 (33.8 and 47.0%), and no. 4 (20.0 and and fatty acid profile. Palm and sunflower oils were chosen 10.6%), respectively, as shown in Table 1. The preference due to their versatile production in Thailand, in addition was in accord with the aroma profiles that extriats no. 1 to their cosmetic application (11). Sunflower oil contains contained more fragrance compounds. Thus, linalool, polyunsaturated fats, triglycerides, and essential fatty acids nerolidol, cis-farnesol, trans-farnesol and ethyl linoleate that maintain the skin barrier by reducing dry skin (13), in a play an important role in the plumeria fragrance. In manner similar to that of palm oil (14). addition, the farnesol derivatives might be considered as An optimized base system, which had a high consistency, the characteristic odour of Thai white plumeria, as they was odourless to virtually odourless, and had a semi-hard were not reported in Plumeria spp. before. surface that allowed easy removal of the flower was preliminary screened from a 1:1 ratio. Two best fat bases Conclusion consisting of a wax and a vegetable oil in a 3:2 ratio were the spermaceti wax and palm oil, and the white beeswax Spermaceti wax and palm oil gave the best enfleurage base and sunflower oil. These bases were further included in this for white plumeria aroma extraction. The aroma profile of the study. Although the base itself had an odour before the extraits and absolutes of Thai plumeria could provide a new aroma absorption process started, we were still able to and much appreciated fragrance that would fulfill the huge obtain the powerful, fresh plumeria odour, particularly in the demand for new oriental flower scents. This class of natural spermaceti wax and palm oil system. However, this system aroma material facilitates more choices of floral notes. The gave less aroma extract than that of the white beeswax use of the old style pomades still provides a challenge and at and sunflower oil (Table 1). Therefore, the enfleurage base the moment these pomades are best reserved for products composed of spermaceti wax and palm oil was shown like decorative candles and cosmetic ointments. to be the most suitable base. This was further supported by the volatiles analysis, which will be discussed later with supportive evidences in terms of the preference results. The Acknowledgements solvent extractions were different in as much as they gave a strongly pungent semi-solid material. Mae Fah Luang University was acknowledged on facility The aroma profiles of the two extraits from spermaceti wax support during this study. The authors gratefully thank for and palm oil, and the white beeswax and sunflower oil Ms. Sasikhan Toso on her assistance in GC-MS analysis. The enfleurages (samples no. 1 and 2), and the two absolutes reviewer was deepest appreciated upon the valuable from ether and n-hexane macerations (samples no. 3 and suggestions that made the article more comprehensive. 4) were comparatively analyzed by GC-MS (8). Aroma compounds were considered identified based on the mass spectra quality of more than 90% of similarity. The References and Notes aroma content was reported based on the peak area of the identified compound. Linalool, nerolidol, cis-farnesol 1 Klaschka U., Kolossa-Gehring M., Envi. Sci. Poolut. Res. 14, and benzyl benzoate were all determined in the aroma 44-52 (2007). preparation (Table 2). The amounts of linalool, nerolidol and 2 Buckley DA., Br. J. Dermatol. 157, 295-300 (2007). cis-farnesol were largest in the extrait obtained from the 3 Narula APS., Chem. Biodiv. 1, 1992-2000 (2004). spermaceti wax and palm oil enfleurage (sample no. 1), 4 Handa SS., Khanuja SPS., Longo G., Rakesh DD., Hydrolytic whereas the amount of benzyl benzoate was highest in the maceration, expression and cold fat extraction, in Extraction extract from ether maceration (sample no. 3). The amount Technologies for Medicinal and Aromatic Plants, Edited by Singh of linalool, nerolidol and benzyl benzoate were in keeping AK., Ed. International centre for science and high technology, with the essential oil distilled from the red coloured flower Trieste (2008). (P. rubra) cultivated in Malaysia (15,16). However, none of 5 Pybus D., Sell C., The Chemistry of Fragrances. Royal Society of the comparative preference evaluations are included in Chemistry, Cambridge (1999). that study. Furthermore, benzoic acid, cis-farnesol, benzyl 6 Lis-Balchin M. Aromatherapy with essential oil. in: Handbook of salicylate, palmitic acid and ethyl linoleate were found Essential oils : Science, Technology, and Applications, Edited by in the extrait enfleuraged by the fat base consisting of Bas¸er KHC., Buchbauer G., Eds. CRC Press, New York (2010). spermaceti wax and palm oil (sample no. 1) but not in the 7 Patin R., Kanlayavattanakul M., Lourith N., IJPS. 5, 161-166 (2009). white beeswax and sunflower oil system (sample no. 2). 8 Zhao C-X., Liang Y-Z., Fang H-Z., Li X-N., J. Chromatogr. A 1096, On the other hand, the white beeswax and sunflower oil 76-85 (2005). 9 Cochrane C-Y., Dubnicka S., Loughin T., J. Sens. Stud. 20, 484-502 system afforded nerol, geraniol, citral, trans-franesol, ethyl (2005). palmitate, octadecenes, and ethyl oleate. The presence of 10 Voraphongsathorn N., Kanlayavattanakul M., Nathakankitkul S., geraniol and palmitic acid in the flower was consistent with IJPS. 4, 73-80 (2008). the red temple tree (15,16). Interestingly, the antibacterial 11 Alvarex AMR., Rodríquez MLG., Grasas y Aceites 51, 74-96 (2000). agents with molluscicidal activity, isoplumericin and 12 Frosch PJ., Peiler D., Grunert V., Grunenberg B., J. Deutschen plumericin (17) were isolated by the n-hexane and ether Dermatol. Gesel. 1, 547-557 (2003). macerations, respectively. 13 Dweck AC., Cosmet. Toiletries 112, 47-64 (1997). Sensory evaluation of the prepared plumeria aroma 14 Keng PS., Basri M., et al., Ind. Crop. Prod. 29, 37-44 (2009). materials was further conducted in terms of quality of 15 Tohar N., Mohd MA., Jantan I., Awang K., Flavour Fragr. J. 21, odour satisfaction and degree of difference from the 859-863 (2006). fresh flower. Preference towards the extriats was higher 16 Tohar N., Awang K., Mohd MA., Jantan I., J. Essen. Oil Res. 18, that from the solvent extractions. Sample no. 1 gained 613-617 (2006). the highest satisfaction (42.6%) with the least difference 17 Hamburger MO., Cordell GA., Ruangrungsi N., J. Ethnopharmacol. from fresh plumeria (38.8%), followed by no. 2 (38.6 33, 289-292 (1991).

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