• TUCHIVES FISHERIES AND MARINE SERVICE 'Iranslation Series No 3984 Composition of the sterol fraction of lipids extracted from oil seeds by E. Tiscornia and G. C. Bertini Original title: Composizione della frazione steroliCa nei lipidi estratti da semi oleosi From: Riv. Ital. Sostanze Grasse 50(Aug.): 251-268, 1973 Translated by the Translation Bureau (MG) • Mültilingual Services Division Department of the Secretary of State of Canada Department of the Environment 41sheries and Marine Service Halifax Laboratory . Halifax, N.S. 1977 30 pages typescript Fd- M .tr- 3 `3 ^ `i DEPARTMENT OF THE SECRETARY OF STATE SECRÉTARIAT D'ÉTAT TRANSLATION BUREAU BUREAU DES ♦,'i^ ï= • TRADUCTIONS ^^•t.' _ MULTILINGUAL SERVICES DIVISION DES SERVICES CANADA DIVISION MULTILINGUES TRANSLATED FROM - TRADUCTION DE INTO - EN Italian English AUTHOR - AUTEUR E. Tiscornia, G.C. Bertini TITLE IN ENGLISH - TITRE ANGLAIS Composition of the sterol fraction of lipids extracted from oil seeds TITLE IN FOREIGN LANGUAGE (TRANSLITERATE FOREIGN CHARACTERS) TITRE EN LANGUE ETRANGÉRE (TRANSCRIRE EN CARACTÉRES ROMAINS) Composizione della frazione sterolica nei lipidi estratti da semi oleosi REFERENCE IN FOREIGN LANGUAGE (NAME OF BOOK OR PUBLICATION) IN FULL. TRANSLITERATE,FOREIGN CHARACTERS. REFÉRENCE EN LANGUE ÉTRANGERE (NOM DU LIVRE OU PUBLICATION), AU COMPLET, TRANSCRIRE EN CARACTÉRES ROMAINS. La Rivista italiana delle sostanze grasse REFERENCE IN ENGLISH - RÉFERENCE EN ANGLAIS The Italian Review of Fatty Substances PUBLISHER- EDITEUR PAGE NUMBERS IN ORIGINAL DATE OF PUBLICATION NUMEROS DES PAGES DANS DATE DE PUBLICATION L'ORIGINAL YEAR ISSUE N0. 251-268 • VOLUME PLACE OF PUBLICATION ANNÉE NUMERO NUMBER OF TYPED PAGES LIEU DE PUBLICATION NOMBRE DE PAGES DACTYLOGRAPfI1EES Italy 1973 50 29 REQUESTING DEPARTMENT TRANSLATION BUREAU NO. MINISTÈRE-CLIENT Environment NOTRE DOSSIER NO 108922,9 BRANCH OR DIVISION TR}1NSLATOR (INITIA LS) Fisheries M. G. DIRECTION OU DIVISION TRADUCTEUR (INITIALES) PERSON REQUESTING DEMANDÉ PAR A. m- Reid ^Jr'4..r.)i i ':a^_ i., r^9.4 .....?4i ll.^:I YOUR NUMBER i C:C• 1iliC'. <, . ..) ' .'.)V VOTRE DOSSIER NO _'^i,.•f C., DATE OF REQUEST 3 3^77 DATE DE LA DEMANDE_ SOS•200-10-6 (REV. 2/66) 7630-21-029-6333 1-doilt3VLI Secretary Secrétariat State d'Étal TRANSLATION BUREAU BUREAU DES TRADUCTIONS MULTILINGUAL SERVICES DIVISION DES SERVICES DiVISION MULTILINGUES . CLIENTS NO. DEPARTMENT DI visioN/URANCH cl iv Ne) DU CLIENT MINISTÉRE DIVISION/DIRECTION VILLE • Environment Fisheries Ottawa VUREAU NO. LANGUAGE TRANSLATOR (MI riALs) , No Du OUREAU LANGUE . TRADUCTEUR (INITIALES) 1089229 Italian M.G. -... Fr cdy ?I ce61,51.F. T RA Us CT 0 0 siTION OF THE STEROL FRACTION OF LIPIDS EXTRACTED FROM OIL SEEDS informeion sainement E. Tiscornia, G.C. Bertini Summary - Within the framework of research undertaken with a view to better characterizing the various kinds of natural vegetable oils and fats for ' human consumption, the Authors in this paper have considered the composition of the sterol fraction of oil seeds commonly used in the Italian oil industry. Based on the results obtained using "coordinated analytical techniques" (TLC and GLC), the Authors found that the qualitative sterol composition of the oils under consideration is generally independent of the variety and region of production. For some seeds, however, these factors seem to have some influence on the quantitaave composition which for some types remains within narrow limits of variability, whereas for others,.the variability shows broader fluctuations. A systematic study of the non-glyceride constituents of vegetable oils was'begun more than ten years ago by Jacini and his group (1-8). The of these and other Authors provided a meaningful picture of the compo- work sition of vegetable oils and demonstrated that this composition is closely related to the species of plant from which the oil is extracted (9-31). Among the various constituents which,.in the present state of our knowledge, are believed to make up the unsaponifiable fraction of a vege- table oil, the sterol fraction is the one that has been most extensively investigated by researchers. SEC 5-25T (6/76) 2 Recent studies appearing in the literature (27-39) have helped 'decisively in the selection and improvement of analytical methods in their various phases, also providing interesting results on the sterol composi- tion of several edible oils. On the present state of researCh,all Authors essentially agree on the analytical method which consists in separating the various unsaponifiable fractions by TLC preparation on plates Of silica gel or alumina; however, two different tendencies exist as regards the GLC analysis of sterols. According to Wolff et al. (38, 40-43) GLC analysis can be carried out directly on free sterols, using stainless steel columns kept at a high ' temperature for •a few days and inactivating the reaction centres of the stationary phase with repeated injections of standard mixtures. However, according to other Authors (35-37, 39, 44-53) it is pre- ferable to convert free sterol into derivations better suited for GLC, such as, for example, trimethylsilyl ethers (TMS). They also suggest the use of special experimental conditions of gas chromatography, such as the use of instruments permitting on-column injection, or quartz injectors and glass columns filled with an acid-washed and silanized support. In this manner, onè obtains optimum conditions for the qualitative and especially for the quantitative determination of the individual sterols, preventing the possibility of breakdown of the sterol molecule, dehydration of the steràls on the column and phenomena of irrèversible adsorption on the active points of the column. It may be noted that the results reported in the literature generally show very significant differences in the composition of the sterol fraction of edible vegetable oils obtained from different species of seeds or fruits, but much less is known about the relationship existing between sterol fraction 3 and variety and/or- place of production of a same species. These factors are known to noticeably affect the fatty acid composition of an oil, and sometimes to cause marked variations. Therefore, we can say that up to now, the possible effect of the abovementioned factors on the composition of the sterol fraction, mainly with respect to different production zones (20, 24, 29, 36-739, 42, 43, 54, 55), has only been taken into account for the various types of oil derived from "Olea europea", but similar experiments have not been carried out on oils obtained from other fruits or seeds (11, 17-19, 28, 29, 33, 36, 56, 57). In fact, only few papers have been published on this subject, a paper by Capella and Losi (32) on the analysis of the sterol fraction obtained from two varieties of "Brassica Napus" seeds (Matador and Regina II), and a paper by Tiscornia and Monacelli (35) on the composition of the sterol fraction of the oil obtained from two different va-rieties of fruit, commonly called seeds, of "Carthamus Tinctorius". The two varieties-were found to contain different levels of linoleic acid. On the basis of the foregoing, we thought that an experimental work investigating the composition of the sterol fraction of lipids extracted from oil-seeds commonly used by the Italian oil industry would be of interest. For each species of seeds used, the production zone was studied and for some samples the different varieties were also taken into account. EXPERIMENTAL METHODS The experiments were conducted on 38 samples of oil extracted from seeds or fruits, belonging to 11 different plant species, of foreign origin with the exception of the seeds of "Vitis vinifera" produced in Italy. 4 32 of these samples were obtained from homogenous, controlled lots of seeds from a single species, free of extraneous seeds, by an industrial extraction method involving pressure and subsequent solvent extraction; two samples, namely one obtained from fruits of "Carthamus Tinctorius", of Spanish origin, and one obtained from seeds of "Brassica Napus", of Canadian origin, were extracted in our laboratory by means of a Soxhlet apparatus with petroleum ether (40-60 °C) following grinding and drying at low temperature. The exact procedure used to extract the remaining four samples, namely three samples of "Carthamus" oil of American origin and one of "Thea" oil of Chinese origin was unknown since the samples were already in the oil state. However, we can say with absolute certainty that these samples were genuine. For all oil samples, the composition of the unsaponifiable sterol fraction was determined using the method of analysis described in N.G.C. Ba III 13 (1971) (58). Essentially the method consists of the following steps: a) preparation of the unsaponifiable fraction; h) separation of the sterol fraction on TLC; c) preparation of the sterol trimethylsilyl derivatives (TMS); d) GLC analysis of TMS. We should point out here that the gas chromatographic determination of each sample were done on two different apparatus simultaneously under the following experimental conditions; 1) Apparatus: Perkin Elmer gas chromatograph, mod. 990, equipped with flame ionization detector and glass injection port; Glass chromatographic column 3 m long with an interior diameter of 1.5 mm; Liquid phase: methylsilicone polymer SE 30 2.5%; . 5 Support: Chromosorb W 100-120 mesh washed .with acids and silanized; o Temperature of the column: 245 C; o Temperature of the evaporator: 280 C; Carrier gas: nitrogen with a flow rate of 25-30 ml/min; Velocity of the paper: 1.25 mm/min. - 2) Apparatus: Fractovap Carlo Erba mod. G 1, equipped with flame ionization detector and on-column injection; • Glass chromatographic column 2.80 m long with an inner diam. of 2 mm; Liquid phase: methylsilicone polymer SE 30 2%; Support: Chromosorb W 100-120 mesh washed with acids and silanized; Temperature of the column: 240°C; o Temperature of the injection block: 280 C; Carrier gas: Helium with a flow rate of 20-25 ml/min; Velocity of the paper: 6"/h.