1321 Sterols and Fatty Acids of Two Caesalpiniaceae J. Miralles o , N. DiallOO, E. Gaydou b and J.M. KornprobsfC ooepartement de Biologie Vegetale, Faculte des Sciences, Universite Cheikh Anta Diop de Dakar, Dakar, senegal; bLaboratoire de Phytochimie, Ecole Superieure de Chimie de Marseille, Av. Escadrille Normandie-Niemen, 13397 Marseille Cedex, France, and coepartement de Chimie, Faculte des SCiences, Universite Cheikh Anta Diop de Dakar, Dakar, senegal TABLE 1 Analytical Data on Cassia hirsuta and C. tora Seed Oils RESULTS AND DISCUSSION The seeds of Cassia hirsuta and C. tora yielded 3.0% and 5.4% oil, respectively. The content and distribution of fatty acids are sum- marized in Table 1. About 15 compounds were identified in the two oils. A high level of unsaturated fatty acids was observed (76.6 and 68.2%) with a prominence of linoleic (58.0 and 44.6%) and oleic acids (13.3 and 21.6%). The cyclopropenoid acids, malvalic and sterculic, were found in small amounts in C. hirsuta, respectively 0.4 and 0.6%, but only a trace of sterculic (0.2%) was found in C. tora. Among the saturated acids, palmitic acid was the were analyzed by GLC and gas liquid chromatography mass spectrometry (GC-MS). GLC was performed with the same apparatus used for FAME, on an OVI fused silica capillary column (0.40 JJffi thick, 25 m X 0.32 mm Ld.). Temperatures were: injector and detector, 280°C; column, 260°C; inlet pressure of hydrogen used as carrier gas was 0.6 bar; split ratio, 1/100. The relative retention times (RRT) for the acetate derivatives were expressed relative to cholesteryl acetate (RRT = 1.00). GC-MS equipment included a Girdel Ribermag RIO-lOB apparatus coupled with a Sidar data computer. The chromatograph was fitted with the same column as described above. Operating conditions were: temperature column was programmed from 250° to 290°C at 5°C/min; helium was used as carrier gas at 0.5 bar; ion source, 150°C; ionizing voltage, 70 eV. 0.2 22.8 0.7 0.2 6.7 21.4 44.6 1.1 1.3 0.2 0.6 5.4 2.6 1.2 0.2 31.8 68.2 Cassia tara 3.0 5.4 2.7 0.4 0.3 16.9 0.6 0.2 2.9 13.3 58.0 3.3 0.8 0.4 1.1 0.8 0.4 0.6 23.4 76.6 Cassia hirsuta Oil content in seeds (%) Unsaponifiable matter (%) Sterol content in oil (%) Fatty acids: 12:0 14:0 16:0 16:1w7 17:0 18:0 18:1w9 18:2w6 18:3w3 20:0 20:1w9 22:0 24:0 Malvalic Sterculic Total saturated Total unsaturated EXPERIMENTAL PROCEDURES Seeds collected in the Dakar area were ground to a powder and extracted with hexane in a Soxhlet apparatus. The solvent was removed at 45°C under reduced pressure and the oil content was determined. Fatty acids study. The fatty acid methyl esters (FAME) were prepared by transesterification of oil with a solution of IN sodium methoxide in methanol and treated with anhydrous methanol saturated with silver nitrate (4). The normal methyl esters and the reaction products from cyclopropene fatty acids were recovered and analyzed by gas liquid chromatography (GLC). GLC was performed with a Carlo-Erba model 4130 gas chromatograph equipped with a flame ionization detector. A carbowax 20M fused silica capillary column (0.40 JJm phase thick- ness, 25 m X 0.32 mm Ld.) was used to separate fatty acid methyl esters. Temperatures were: injector and detec- tor, 230°C; column 175°C; inlet pressure of hydrogen used as carrier gas was 0.5 bar; split ratio, 5/100. A sample of Sterculia foetida seed oil was used as reference. Sterols studies. Oils were saponified by KOH/EtOH with usual workup and unsaponifiable matters were recovered by n-hexane. Sterols were extracted by digitonin complexation and recrystallized in methanol. The sterol mixture was separated by preparative thin layer chromatography (TLC) on silica gel (0.5 mm thick), developped four times with C 6 H 6 /MeOH (98:2, v/v) into two fractions, one containing mainly 1\-5 sterols and another consisting mainly of 1\-7 and saturated sterols. The two fractions were purified by another TLC (0.2 mm thick) under the same conditions and acetylated with AczO/Py at room temperature overnight. The acetates Previous workers have demonstrated that the seed oil of Cassia grandis, belonging to the Leguminoseae family, contains small amounts of sterculic and malvalic acids (1). In a preliminary study on sterol fractions of some Cassia species we found a complex composition with the presence of many 1\-7 sterols (2). The presence of these different compounds, which are quite rare in higher plants, led us to investigate the lipid composition of two Senegalese Cassia seed oils, Cassia hirsuta and C. tora. Cassia hirsuta L. is an undershrub of 2-3 m height with yellow flowers; leaves, stems and fruits are very long pilose or hirsute. Native to tropical America, it has been introduced into Africa (3). Cassia tora L. is an undershrub of 1-3 m height with yellow flowers in pairs or singly, and is widespread in tropical areas (3). Seed oils of Cassia hirsuta and C. tora L., two Caesalpiniaceae belonging to tbe Leguminoseae family, contain small amounts of malvalic and sterculic acids. Sterol composition was determined by capillary GLC and GC/MS; 15 sterols were identified. In addition to the usual 1\-5 sterols, high levels of 1\-7 sterols and small amounts of saturated sterols were found. JAOCS, Vol. 66, no. 9 (September 1989)