College of Science and Technology, Nihon University (1-8 Kanda Surugadai, Chiyoda-ku, Tokyo , JAPAN) 2

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1 Journal of Oleo Science Copyright 2010 by Japan Oil Chemists Society Triterpene Alcohol and Fatty Acid Composition of Shea Nuts from Seven African Countries Toshihiro Akihisa 1, Nobuo Kojima 1, Naoko Katoh 1, Yuki Ichimura 1, Hirohisa Suzuki 1, Makoto Fukatsu 1, Steven Maranz 2 and Eliot T. Masters 3 1 College of Science and Technology, Nihon University (1-8 Kanda Surugadai, Chiyoda-ku, Tokyo , JAPAN) 2 Department of Microbiology and Immunology, Weill Medical College, Cornell University (1300 York Ave, New York, NY, U.S.A.) 3 World Agroforestry Centre (ICRAF) (PO Box Nairobi, KENYA) Abstract: The content and composition of triterpene alcohol fractions of the non-saponifiable lipids (NSL) along with the fatty acid composition of the kernel fats (n-hexane extracts) of the shea tree (Vitellaria paradoxa; Sapotaceae) were determined for 36 samples from seven sub-saharan countries: Cote d Ivoire, Ghana, Nigeria, Cameroun, Chad, Sudan, and Uganda. The fat content of the kernels, proportion of NSL in the fats, and triterpene alcohols in the NSL are in the range of 30 54, 2 12, and 22 72, respectively. The triterpene alcohol fractions contained α-amyrin (1), β-amyrin (2), lupeol (3), and butyrospermol (4) as the major constituents along with minor or trace amounts of ψ-taraxasterol (5), taraxasterol (6), parkeol (7), 24-methylene-24-dihydroparkeol (8), 24-methylenecycloartanol (9), dammaradienol (10), and 24-methylenedammarenol (11). Fatty acid composition is dominated by stearic (28 56 ) and oleic (34 61 ) acids. Shea butters from West African provenances contained in general higher levels of triterpene alcohols and stearic acid than those from East African provenances. Both stearic acid and total triterpene alcohol contents were significantly correlated to the latitude and elevation of the source population, indicating that higher levels of these compounds are found at higher ambient temperatures. Key words: Vitellaria paradoxa, shea nuts, triterpene alcohol, fatty acid 1 INTRODUCTION The shea tree Vitellaria paradoxa C. F. Gaertn.; synonyms Butyrospermum paradoxum C. F. Gaertn. Hepper, Butyrospermum parkii G. Don Kotschy; family Sapotaceae is indigenous to the savanna belt extending across sub-saharan Africa north of the equator, ranging from Mali in the west to Ethiopia and Uganda in the east extending from 16 W to 34 E longitude and 1 N to 15 N latitude 1-4 Fig. 1. The most valued product of shea tree is shea fat shea butter, the fat extracted from the kernels. Processed shea fat is used primarily as a cocoa butter additive in chocolate manufacture 1. These properties are due to the structure of its component triglycerides. In addition, shea fat is increasingly popular in skin care products and cosmetic product formulations, in part due to the unusually high level of non-saponifiable lipid NSL constituents in the fat 5. The main NSL constituents of shea fat have been reported to be triterpene alcohols such as α-amyrin 1, β-amyrin 2, lupeol 3, and butyrospermol Fig. 2. Since naturally occurring triterpene alco- hols and their derivatives exhibit a variety of biological activities including anti-inflammatory, antitumor, chemopreventive, and antimycobacterial activities 10-14, we wished to explore and determine whether significant differences in qualitative and quantitative compositions exist for the NSL of shea fat among widely dispersed V. paradoxa populations. We now report, in this paper, the contents and composition of the triterpene alcohol fractions from the NSL and fatty acid composition of kernel fats obtained from 36 shea nut samples from seven sub-saharan countries, and observations on the relation between the triterpene alcohol constituents and fatty acid composition. 2 EXPERIMENTAL 2.1 Materials and chemicals Collection of shea nut samples was undertaken by one of the authors E.T.M. using a standard collection protocol Correspondence to: Toshihiro Akihisa, College of Science and Technology, Nihon University, 1-8 Kanda Surugadai, Chiyoda-ku, Tokyo , JAPAN akihisa@chem.cst.nihon-u.ac.jp Accepted January 13, 2010 (received for review November 11, 2009) Journal of Oleo Science ISSN print / ISSN online 351

2 T. Akihisa, N. Kojima, N. Katoh et al. Table 1 Longitude, Latitude, and Elevation of the Sites of Shea Nuts Collection Fig. 1 Map of Africa Showing Sites ( ) of Sampled Shea Nuts in Relation to the Species Distribution Range that Extends from 16 W to 34 E Longitude and 1 N to 15 N Latitude. Fig. 2 Structures of 11 Triterpene Alcohols Identified in the Non-saponifiable Lipid Fraction of Shea Kernel Fat. developed by one of the authors S.M. under contract to World Agroforestry Centre ICRAF, with subsequent laboratory treatment and sample preparation and transfer to Nihon University supported by ICRAF in Samanko Mali, Yaoundé, and Nairobi. Shea nut samples were collected during the 2006 shea season May through July from healthy mature trees at 36 sites of seven countries in Africa Table 1 and Fig. 1. These included two sites in Cote d - Ivoire, one in Ghana, 14 in Nigeria, three in Cameroun, eight in Chad, two in Sudan, and six in Uganda. The nut samples were numbered sequentially based on the longitude of the collection site, beginning with the westernmost site. Standard fatty acid methyl esters were purchased from Sigma-Aldrich Japan Co. Tokyo. Country Longitude Latitude Elevation (m) Cote d Ivoire 1 W 7º N 10º W 5º 6 37 N 9º Ghana 3 W 0º N 6º Nigeria 4 E 4º N 11º E 5º 18 7 N 10º E 5º N 9º E 6º N 10º E 6º N 9º E 6º N 8º E 6º N 10º E 6º N 10º E 7º 27 9 N 9º E 8º N 8º E 9º N 7º E 11º N 9º E 12º 29 6 N 9º E 12º N 9º Cameroun 15 E 10º N 5º E 11º 2 56 N 5º E 13º N 7º Chad 21 E 14º N 9º E 15º N 9º E 15º N 9º E 16º 9 17 N 8º E 16º N 9º E 17º 4 44 N 8º E 17º N 9º E 18º 2 19 N 9º Sudan 29 E 28º N 7º E 30º N 6º Uganda 31 E 33º N 2º E 33º 13 0 N 3º E 33º 15 7 N 2º E 33º N 2º E 33º N 2º E 33º N 2º

3 Triterpene Alcohol and Fatty Acid Compositions of Shea Fat 2.2 Kernel fat content Whole nuts were oven-dried at 60 over 72 h and decorticated. Kernels were crushed in a mortar and finely ground in a coffee mill. Pulverized samples were weighed and extracted with n-hexane under reflux three times for 3 h each. Fat percentage was determined gravimetrically by dividing extracted total fat by kernel weight after evaporation of solvent. 2.3 Saponification and separation of triterpene alcohol fraction The n-hexane extract was subjected to alkaline hydrolysis 5 KOH in MeOH, reflux, 3 h followed by diisopropyl ether extraction, yielding a neutral NSL fraction. The percentage of NSL in the fat was determined gravimetrically by dividing the weight of the extracted NSL by the n-hexane extract weight after evaporation of the solvents. The NSL fraction was further fractionated on cm plates coated with a 0.5 mm layer of Silica gel G60 Merck & Co., Inc.. The sample 25 mg was applied to the plate and developed with n-hexane ethyl acetate EtOAc 8:2 for 1 h. The plate was sprayed with 0.1 rhodamine 6G solution in MeOH and observed under UV light. The portion of the TLC plate containing triterpene alcohols R f 0.55 was cut off and quantitatively extracted with n-hexane EtOAc 7:3. The percentage of triterpene alcohols in the NSL was determined gravimetrically by dividing the extracted triterpene alcohol fraction by the NSL fraction. The triterpene alcohol fraction was acetylated in acetic anhydride and pyridine overnight at room temperature. The acetate fraction was used for GLC analysis and for isolation of individual components by argentation TLC followed by preparative HPLC. 2.4 Isolation and identifi cation of triterpene alcohols Shea nut sample no. 12 from Nigeria was selected for in-depth analysis. The triterpene alcohol fraction 2704 mg gave an acetate fraction of 2887 mg after acetylation. The acetate fraction was subjected to preparative argentation TLC 6 developing solvent: cyclohexane EtOAc, 95:5, giving five fractions: fractions A 1152 mg; R f value 0.90, B 168 mg; 0.81, C 789 mg; 0.79, D 47 mg; 0.73, and E 48 mg; A portion of fraction A 30 mg was loaded onto a preparative reversed-phase HPLC column TSK ODS-120A, 25 cm 10 mm i.d.; Toso Co., Tokyo and eluted with MeOH flow rate: 3.0 ml/min, yielding α-amyrin acetate 1a; 18.2 mg; retention time t R 50.8 min 6 and β-amyrin acetate 2a; 3.1 mg; t R 46.8 min 6. A portion 77 mg of fraction B, separated using preparative HPLC as above, gave butyrospermol acetate 4a; 2.9 mg; t R 32.4 min 6, ψ-taraxasterol acetate 5a; 2.8 mg; t R 55.6 min 9, 14, and taraxasterol acetate 6a; 1.4 mg; t R 46.8 min 9, 14. Preparative HPLC as above for a portion 74 mg of fraction C gave lupeol acetate 3a; 10.0 mg; t R 30.7 min 6 and 4a 10.3 mg. Fraction D, likewise on preparative HPLC, yielded 3a 7.1 mg, parkeol acetate 7a; 1.6 mg; t R 47.2 min 6, 24-methylene-24-dihydroparkeol acetate 8a; 2.1 mg; t R 49.2 min 9, 15, and 24-methylenecycloartanol acetate 9a; 5.2 mg; t R 76.5 min 6. Finally, preparative HPLC of fraction E gave 8a 3.8 mg, dammaradienol acetate 10a; 3.1 mg; t R 28.4 min 7, and 24-methylenedammarenol acetate 11a; 10.3 mg; t R 30.0 min 7. The structures of these compounds are shown in Fig. 2. Identification of 11 triterpene alcohols was established for the acetyl derivatives, 1a 11a, by comparison of their 1 H NMR and MS data with those of corresponding compounds cited in the literature 6, 7, 9, 14, Measurement of triterpene alcohol fractions Analysis of the triterpene alcohol fractions, as the acetyl derivatives, was performed by GLC using a Shimadzu GC- 17A series gas chromatograph equipped with a FID. The chromatograph was fitted with a DB-17 fused-silica capillary column 30 m 0.31 mm i.d., 0.25 μm film thickness, J&W Scientific, Inc., Folsom, CA, U.S.A.. The column was operated isothermically at 275 with N 2 64 ml/min; split ratio 33:1 as a carrier gas. Injection and detector temperatures were 300. The retention times established for the reference triterpene alcohols isolated from sample no. 12 Nigeria, described above, were used to identify the triterpene peaks for all samples. Relative retention times Rt R were calculated with reference to an internal standard, cholesterol acetate t R 17.5 min. The compounds and Rt R values were as follows: 1a 1.91, 2a 1.69, 3a 2.00, 4a 1.64, 5a 2.46, 6a 2.60, 7a 1.77, 8a 1.95, 9a 2.00, 10a 1.67, and 11a The relative percentage of each triterpene alcohol was obtained by dividing the individual peak area by the sum of all peak areas obtained for triterpene alcohols using an integrator. 2.6 Preparation and analysis of fatty acid methyl esters A few drops of sulphuric acid 0.15 μl were added to a solution of the n-hexane extract 50 mg in MeOH 50 ml, and the mixture was refluxed for 3 h. The solvent was evaporated on a rotary evaporator and the methyl esters thus prepared were extracted with diethyl ether 3 times after the addition of water. The combined ether extracts were washed with water until neutral, dried with anhydrous sodium sulphate, filtered and taken down to dryness using a rotary evaporator. Fatty acid methyl esters were analyzed with a Shimadzu GC-2014 series gas chromatograph equipped with a FID. The chromatograph was fitted with a fused-silica capillary column 50 m 0.25 mm i.d., 0.25 μm film thickness designed for FAME applications Varian, Inc., Palo Alto, CA, U.S.A.. The column was operated at 220 with He as a carrier gas flow rate 157 ml/ min; split ratio 150:1. Detector and injection temperatures were 300. Identification of fatty acids was done by the use of standards. The relative percentage of each fatty acid 353

4 T. Akihisa, N. Kojima, N. Katoh et al. was obtained by dividing the peak area of the fatty acid by the sum of all fatty acid peak areas using an integrator. 2.7 Data analysis Data was analyzed statistically using XLSTAT 2009 Addinsoft, New York, NY, U.S.A.. Regression analysis and Spearman correlations were used to examine data trends and to evaluate the relationship between chemical composition and geographic parameters. 3 RESULTS AND DISCUSSION 3.1 Kernel fat content As shown in Table 2, the fat content of shea kernels investigated ranged from 29.7 sample no. 15 from Cameroun to 53.7 sample no. 12 from Nigeria with a mean value of No striking regional difference in fat content was observed among the samples examined except those from Cameroun, which showed low fat percentage with a mean value of Low kernel fat content for Cameroon shea is consistent with that reported in the literature NSL content of kernel fat The NSL content of kernel fats ranged from 2.4 nos. 34 and 36 from Uganda to 11.6 no. 16 from Cameroun with a mean 6.2 Table 2. Whereas considerable variation was observed in the NSL content within a country e.g., for the samples from Nigeria, shea samples from western area of the shea belt Cote d Ivoire, Ghana, Nigeria, and Cameroun exhibited higher NSL content than those from eastern area Sudan and Uganda. 3.4 Triterpene alcohol composition The composition of triterpene alcohol fractions as acetyl derivatives from the NSL of 36 shea samples are shown in Table 3. Among the eleven GLC peaks observed, Rt R 1.64, 2.46, and 2.60 were respectively identified as butyrospermol 4, ψ-taraxasterol 5, and taraxasterol 6. Some compounds co-eluted in the same peak: Rt R 1.69 was identified as β-amyrin 2 and dammaradienol 10, of which the former was estimated to be the predominant component based on the isolation experiment done for shea sample no. 12 from Nigeria, described above. In a similar manner, the peak with Rt R 1.77 was estimated to consist of parkeol 7 and 24-methylenedammrenol 11, while Rt R 1.91 consisted of α-amyrin 1 and 24-methylene-24-dihydroparkeol 8, and Rt R 2.00 consisted of lupeol 3 and 24-methylenecycloartanol 9. Taking these into consideration, the acetylated triterpene alcohol fractions of the shea samples examined were shown to contain α-amyrin as the predominant component followed by lupeol , butyrospermol , and β-amyrin , accompanied by minor or trace amounts of compounds 5, 6, 8, 9, 10, 11, and several unidentified compounds. An example of the GLC chromatogram for the triterpene alcohol fraction analyzed as acetyl derivative obtained from shea nut sample no. 1 from Cote d Ivoire was shown in Fig. 3. Both the total triterpene alcohol content and the relative percent composition of individual triterpene alcohols showed significant correlations with geographic variables Table 4. Higher total triterpene levels are associated with higher latitude and lower elevation, both of which reflect hotter temperatures. Among individual triterpene alcohols, butyrospermol 4 most closely tracked the hot temperature trend. Both α-amyrin 1 and β-amyrin 2 showed an inverse tendency, correlating significantly with 3.3 Triterpene alcohol content Triterpene alcohol fractions in the NSL ranged from 22.4 no. 35 from Uganda to 71.7 no. 12 from Nigeria with a mean 48.1 Table 2. As has been observed in the NSL content described above, the proportion of triterpene alcohols in the NSL also varied considerably within a country e.g., for the Nigerian samples. Shea samples from western area of shea belt Cote d Ivoire, Ghana, Nigeria, and Cameroun had a tendency to contain more triterpene alcohols in the NSL than those from eastern area Table 2. Shea samples from Chad, located in the central area of shea belt, showed intermediate levels of triterpene alcohols compared to Sudan and Uganda to the east and Nigeria, Ghana, and Cote d Ivoire to the west. Fig. 3 GLC Chromatogram of the Triterpene Alcohol Fraction (analyzed as the acetyl derivative) Obtained from Shea Nut Sample No. 1 [see footnote b) of Table 3 for the names of compounds (1) (11)]. 354

5 Triterpene Alcohol and Fatty Acid Compositions of Shea Fat Table 2 Content of Shea Kernel Fats, Content of NSL in the Fat, and Contents of Triterpenes in the NSL and in the Fat Country Fat content of shea kernel ( ) NSL in the fat ( ) Triterpenes in the NSL ( ) Triterpenes in the fat ( ) Cote d Ivoire SD a) Ghana Nigeria Cameroun Chad Sudan Uganda Mean (SD) 42.4 (6.1) 6.2 (2.2) 48.1 (13.7) 3.1 (1.5) a) Standard deviations (n=3) for the content of shea kernel fat, content of NSL in the fat, and content of triterpenes in the NSL determined for sample no

6 T. Akihisa, N. Kojima, N. Katoh et al. Table 3 Composition of Triterpene Alcohol Fractions from the Non-saponifiable Lipid Fractions of the Kernel Fats of 36 Shea Nut Samples Determined by GLC Rt R a) of individual components b) Composition (4) (2) >> (10) (7) > (11) (1) >> (8) (3) >> (9) (5) (6) Shea nut sample Cote d Ivoire SD c) Ghana Nigeria Cameroun Chad , Sudan Uganda a) Rt R =Relative retention times for the acetyl derivatives. Retention time for cholesterol acetate (17.5 min) is taken as b) α-amyrin (1), β-amyrin (2), lupeol (3), butyrospermol (4), ψ-taraxasterol (5), taraxasterol (6), parkeol (7), 24-methylene-24- dihydroparkeol (8), 24-methylenecycloartanol (9), dammaradienol (10), and 24-methylenedammarenol (11). c) Standard deviations (n=3) deteremined for sample no

7 Triterpene Alcohol and Fatty Acid Compositions of Shea Fat Table 4 Spearman Correlations (r s ) between Geographic Parameters and the Dominant Fatty Acids, Total Triterpene Alcohol Content, and the Four Major Triterpenes in Shea Fat, based on Analyses of 36 Shea Nut Samples Stearic acid (18:0) Oleic acid (18:1) Triterpenes in fat ( ) α-amyrin (1) a) Lupeol (3) b) Butyrospermol (4) β-amyrin (2) c) Variables r s p-value r s p-value r s p-value r s p-value r s p-value r s p-value r s p-value Latitude < < Longitude < < < Elevation < < < a) Eluted with 24-methylene-24-dihydroparkeol (8). b) Eluted with 24-methylenecycloartanol (9). c) Eluted with dammaradienol (10). higher elevation and lower latitude, indicating increasing relative abundance with cooler temperatures in comparison to other triterpenes. It should be noted that the absolute quantities of α-amyrin 1 and β-amyrin 2 are still higher in lowland shea populations, due to the substantially higher total triterpene alcohol content. Lupeol 3 had no significant association with any of the parameters evaluated. 3.5 Fatty acid composition Eighteen saturated and unsaturated fatty acids were characterized in the shea kernel fat in this study Table 5 : myristic myr, 14:0, pentadecanoic pen, 15:0, palmitic pam, 16:0, palmitoleic Δpam, 16:1, n 7, heptadecanoic ept, 17:0, stearic ste, 18:0, elaidic ela, 18:1, trans-9, oleic ole, 18:1, n 9, cis-vaccenic vac, 18:1, cis-11, linolelaidic lnl, 18:2, trans-9, trans-12, linoleic lin, 18:2, n 6, linolenic lnn, 18:3, n 3, arachidic ach, 20:0, eicosenoic eic, 20:1, n 9, behenic ben, 22:0, brassidic bra, 22:1, trans-13, erucic eru, 22:1, n 9, and lignoceric lig, 24:0 acids. Among these, the predominant fatty acid was oleic acid followed by stearic , linoleic , palmitic , and arachidic tr 1.8 acids. Stearic and oleic acids together accounted for of the fatty acids in most of the samples. This fatty acid profile range is consistent with previous studies reporting values for 432 shea samples in 42 populations in 10 African countries 2 and for 158 shea samples in 20 populations in four countries 4. The fatty acid profiles of the shea samples analyzed show a strong geographic trend, paralleling what we found with the triterpene alcohols. Stearic acid content has a significant positive correlation with higher latitude, and a significant negative correlation with both increasing elevation and easterly longitude Table 4. Oleic acid shows the inverse relationship. Since these two fatty acids are the dominant components of shea fat and are represented by relative percentage data, an increase in one is expected to be accompanied by a decrease in the other. However, it appears that it is climate that primarily determines the bal- Fig. 4 Correlation between the Dominant Fatty Acids and Total Triterpene Alcohol Content of Extracted Fat in 36 Shea Nut Samples. a) Stearic acid is positively correlated (R 2 =0.29, p=0.001) with triterpene alcohols, while b) oleic acid is negatively correlated (R 2 =0.30, p < 0.001) 357

8 T. Akihisa, N. Kojima, N. Katoh et al. Table 5 Fatty Acid Composition ( ) of the n-hexane Extracts of 36 Samples of Shea Nuts a) Composition Rt b) R of individual myr pen pam Δpam ept ste ela ole vac lnl lin lnn ach eic beh bra eru lig components 14:0 15:0 16:0 16:1 17:0 18:0 18:1 18:1 18:1 18:2 18:2 18:3 20:0 20:1 22:0 22:1 22:1 24: Shea nut sample Cote d Ivoire 1 tr 3.4 tr tr tr tr 3.3 tr tr 0.1 SD c) Ghana tr tr 0.1 Nigeria 4 tr tr 3.0 tr tr tr tr 2.9 tr tr tr tr 3.5 tr tr 3.3 tr tr tr 3.0 tr tr tr tr tr Cameroun 15 tr 4.8 tr tr tr tr tr tr 0.1 Chad 21 tr tr tr tr 22 tr tr 4.1 tr tr tr tr 3.9 tr tr tr 3.2 tr tr tr 3.9 tr tr tr 3.8 tr tr tr 4.0 tr tr tr 4.7 tr Sudan tr tr 37.3 tr tr tr Uganda tr tr tr tr 32 tr 3.8 tr tr tr tr 3.7 tr tr tr tr 34 tr tr tr 3.9 tr tr tr tr 3.8 tr tr 0.1 a) Percentage composition of fatty acid determined by GLC as the methyl ester derivative. tr=trace amounts. b) Rt R =Relative retention times for the methyl ester derivatives. Retention time for methyl palmitate (45.4 min) is taken as c) Standard deviations (n=3) detemined for the fatty acid composition for sample no

9 Triterpene Alcohol and Fatty Acid Compositions of Shea Fat ance between the two major fatty acids, since the temperature gradient across the shea savanna belt increases with north latitude and decreases with elevation. The data from this study show that the stearic acid and triterpene alcohol contents of shea fat generally increase together, while oleic acid content correlates negatively with total triterpene alcohols Fig. 4, again reflecting the underlying link with temperature. The data analysis also shows a significant longitudinal trend Table 4. This could be interpreted as reflecting genetic differences in keeping with the current taxonomic division into two subspecies, V. paradoxa ssp. paradoxa in the West and V. paradoxa ssp. nilotica in the East 2, 4. Among the shea nut samples investigated, those nos from ssp. nilotica, collected in Sudan and Uganda, are compositionally distinct from the West African types. The former possess consistently high oleic acid and low stearic acid fat. However, samples from Sudan, Chad, and Cameroun show intermediate values that indicate a continuum across the shea belt, rather than two distinct populations Table 5. A study of genetic markers in shea populations in eight countries covering most of the Vitellaria range showed that genetic distances between populations were strongly correlated to geographic distances, showing an east-west incremental transition 16. The longitudinal effect seen in our results could reflect this gene flow; however, it seems clear from the strong effect of elevation on shea lipid composition that temperature plays a major determinant role. 4 CONCLUSION In this study, the kernel fats n-hexane extracts of 36 shea nut samples from seven sub-saharan countries were analyzed for the triterpene alcohol constituents in the nonsaponifiable lipid fractions and for fatty acid constituents. Consistent with earlier findings, the kernel fats contained unusually high levels of triterpene alcohols, ca , although the values we measured were not as high as reported previously by di Vicenzo et al. 4. Four compounds, α-amyrin 1, β-amyrin 2, lupeol 3, and butyrospermol 4, constituted the major triterpene alcohols for all the nut samples investigated. Fatty acid composition is dominated by oleic and stearic acids. The triterpene alcohol content of shea nuts generally increased with increasing stearic acid percentage, while decreasing with greater oleic acid content. This trend reflects an association of shea lipid composition with climate, underscored by significant correlations with both latitude and elevation. Shea nuts from Ugandan provenances had substantially lower triterpene alcohol content and higher oleic acid percentages than those from West Africa, with intermediate values seen in central African provenances. The results of this study will be of value for further utilization of shea kernel fat shea butter from specific origins according to product application in the cosmetic and pharmaceutical fields in the future. ACKNOWLEDGMENTS Sampling of Vitellaria paradoxa across Africa in support of this study was implemented during 2006 with funding from the World Agroforestry Centre ICRAF with additional support from the Common Fund for Commodities CFC from 2004 to 2007 under project CFC/FIGOOF/23 Improving Product Quality and Market Access for Shea Butter Originating from Sub Saharan Africa. Sampling in Ghana, Nigeria, Cameroon, Sudan, and Uganda was undertaken with the support of Bréhima Kone and Momoudou Dia of ICRAF, by Nafan Diarrassouba in Côte d Ivoire, by Victor Keouna and Antcha Mady, and Aliouda Limane Worgué in Chad, in collaboration with the Bup Nde Divine University of Ngaoundéré in Cameroun and Gordon Wagner of Lulu Works in Sudan. References 1 Masters, E.T.; Yidana, J.A.; Lovett, P.N. Reinforcing sound management through trade: shea tree products in Africa. Unasylva No. 219, 55, Maranz, S.; Wiesman, Z.; Bisgaard, J.; Bianchi, G. Germplasm resources of Vitellaria paradoxa based on variations in fat composition across the species distribution range. Agrofor. Sys. 60, Maranz, S.; Kpikpi, W.; Wiesman, Z.; de Saint Sauveur, A.; Chapagain, B. Nutritional values and indigenous preferences for shea fruits Vitellaria paradoxa Gaertn. F. in African agroforestry parklands. Econ. Bot. 58, di Vincenzo, D.; Maranz, S.; Serraiocco, A.; Vito, R.; Wiesman, Z.; Bianchi, G. Regional variation in shea butter lipid and triterpene composition in four African countries. J. Agric. Food Chem. 53, Alander, J. Shea butter a multifunctional ingredient for food and cosmetics. Lipid Technol. 16, Itoh, T.; Tamura, T.; Matsumoto, T. Sterols, methylsterols, and triterpene alcohols in three Theaceae and some other vegetable oils. Lipids 9, Itoh, T.; Tamura, T.; Matsumoto, T. 24-Methylenedammarenol: a new triterpene alcohol from shea butter. Lipids 10, Peers, K.E. The non-glyceride saponifiables of shea butter. J. Sci. Fd. Agric. 28,

10 T. Akihisa, N. Kojima, N. Katoh et al. 9 Itoh, T.; Uetsuki, T.; Tamura, T.; Matsumoto, T. Characterization of tritierpene alcohols of seed oils from some species of Theaceae, Phytolaccaceae and Sapotaceae. Lipids 15, Akihisa, T.; Yasukawa, K.; Kimura, Y.; Takase, S.; Yamanouchi, S.; Tamura, T. Triterpene alcohols from camellia and sasanqua oils and their anti-inflammatory effects. Chem. Pharm. Bull. 45, Fernández, M.A.; de las Heras, B.; García, M.D.; Sáenz, M.T.; Villar, A. New insights into the mechanism of action of the anti-inflammatory triterpene lupeol. J. Pharm. Pharmacol. 53, Akihisa, T.; Yasukawa, K. Anti-inflammatory and antiallergic properties of triterpenoids from plants. in Biomaterials from Aquatic and Terrestrial Organisms Fingerman, M.; Nagabhushanam R. ed., Science Publ. Enfield. pp Akihisa, T. Anti-inflammatory, antitumor, and chemopreventive effects of triterpenes from plants and fungi. Oleoscience 7, Akihisa, T.; Yasukawa, K.; Oinuma, H.; Kasahara, Y.; Yamanouchi, S.; Takido, M.; Kumaki, K.; Tamura, T. Triterpene alcohols from the flowers of Compositae and their anti-inflammatory effects. Phytochem. 43, Itoh, T.; Tamura, T.; Matsumoto, T. 24-Methylenelanost en-3β-ol, new triterpene alcohol from shea butter. Lipids 10, Fontaine, C.; Lovett, P.N.; Sanou, H.; Maley, J.; Bouvet, J-M. Genetic diversity of the shea tree Vitellaria paradoxa C.F. Gaertn, detected by RAPD and chloroplast microsatellite markers. Heredity 93,