A total characterisation of Moringa oleifera_ Malawi seed oil CARATTERIZZAZIONE DELLOLIO DI SEM! DI MORINGA OLEIFERA MALAWI E stato prodoto oli da! semi di Moringa olaifara Malawi usando we diversi “sistem: pressione a freddo (CP), astrazione cor n-esano (M) ed asirazione con ‘Una miscala 60:50 a clorotormio:metanoto (OM). Lolio prodotlo angava dai 25.1% ‘con (CP) 21 43.4% con (CM). Sono stati determinali densila, indice dk rifrazione, ‘colore, punto di fumo, viscasita, cia , numero di saponificazione, numero &i Jodio, metilssten degit aciel grass, tera, tocclerol (per HPLC). numero a perossicl. E%, a £82 nm © 270 mm @ la sonsibillé alossidazione con i! metodo Rrancimat, ‘S18 trovaio che Folio contiene alt live ai acid grassi insaturi, ed In «odo ‘spaciale oleico (ole il 678098). | principal! acid) saturi sono i bohenico (oli 68796) 0 10 siearico (ol il 586%). & stato ancho detorminato un allo cOnlenuto 4i f-sitosterola (superiore a) 47,108), campasierolo (sup. 2 2383%) @ ‘stigmasteroia (sup. al 17.40%). a ‘y= 2 butocoteroll sono presenti rispetivamenta ‘con To seguenti quantilé: 2269, 7147 @ 21657 mg/kg. Nl periodo ai Induzione (2 120°O) dell'ol0 6 Moringa olaiera @ stato idotto dal 49% a! 74% dopo. emucilagginszione. Lolo presenta alta stablila allossidazione. In conttonto con Yolo of ova, Folio dei semi di Moringa oleitera presenta una maggiore stabil, up Illa inféviore of insaturazione & camposiziona acigiea simile (eccettvati Ci:2 0 C183). |, The olifrom the soods of Moringa oleitra Malawi was produced using three iterent wats. Cold pressure (CP), extraction with mnexane (F) and extraction “with ar determined. ture of chlorotorm:methanol (50:80) (CM). The oil produced ranged tom 2511 (CP) to 414% (CM). The density, refracive index, colour, smoke point, Viscosity, aciclly, saponification valve, ladine valve, lay acid methyl esters, sterols, tocopherols (by HPLC), peroxide value, Eat 232 nm and 270 nm ang {he susceplbilly 0 oxidation measured by the Aaseimat method were ‘The ol as found to contain high levels of unsatursted fatty acids, especially oleic (up to 67.80%), Tho dominant salurated acid was behanic (up to 631%) and Stearic (Up to 586%), The cil was also found to contain high lavels of Besitosterel (up to 47-10%), campesterol (up lo 2383%) and stiamasiorol (up to J. TSAKNIS, S. LALAS \V. GERGIS, V. SPILIOTIS DEPARTHENT OF FOOD TECHNOLOGY TECHNOLOGICAL EDUCATIONAL STITUTE EL) INTRODUCTION hhe Moringaceae family consists of 10 [25] or 12 [20] spe- ies which belong to only one genus called Moringa. ‘A Moringa species are native fo India from where they have been introduced into many warm countries [24]. Morton {20} reported that the most cammon species are Moringa peregrina (ors fori (yn. M. aptera Gaertn. M, arabica (Lam) Pers., Moringa zeylanica Sieb,: Balanus myrepsica (Stack), Mosinga stenopetala Culod, Moringa borziana Mattei, Morin- ga longituba Engl, Moringa concanensis Nimmo, Moringa ovalifolia Dinter and A. Berger, Moringa drouhardi, Moringa hildebranti (4. ‘The best known and most widely distributed species is Moringa oleifera (syn. M. pterygosperma Gaarin.) {20, 24) which is @ native of the Western [18] and subrHimalayan tract {24}, India and other courtries of Asia (15, 20, 24), Africa (1, 3, 4, 20], Middle East (15], Philippines, Cambodia, Central “America, Northern South America and the Caribbean Islands (20), The tree ranges in height trom § to 10 m and sometimes even 15m {20}, Sengupta etal. [24}, Morton [20] and Jami Son [14] repories that the tree grows rapiaty even in poor soit and is lite affected by drought. The leaves. tlowers, fruits (which are called "pods" and roots of the tree are used as vegetables {15,20, 23,24), while the trunkis used in the paper industry [18, 271. The fruits are usually 25 to 45 cm long although Ramachadran etal. (28}teported fruits up to 120.6m in length. They contain about 20 seeds {24} which are globu- Jag, about 1 em in diameter, 3-winged with wings produced at the base ofthe apex, 2-25 em long, 040.7 cm wide and scar ious 23}. Sengupta etal [24] reported that the seeds are three- angled and on average weigh about 03 g withthe kernel bring {740%}. and é-locopherole were detected up to lovels of 2269, 71.47 and 21657 malkg respectively. The induction period (at 120°C) at Moringa oleifera ‘seed oll was reduced from 49% 10 749% alter degumming. Moringa oll snowed high stabilly to oxidative rancidiy. Compared to olive ol, Moringa oleifra seed ‘il showed highor stably, lower degree of unsaturation and similar fay acid composition (apart tom C1B:2 and C189) Ing 70-75% of the weight. Ibrahim et al. [12] reported that the oll content and its properties show a wide variation depend- ing mainly on the spacies and the environmental conditions, “There are a few known varieties. They can be distinguished largely by the color and the size of the fruit. Some of therm are reported by Morton [20], Anonymous (1], Ramachadran ot al. [20]. These are Jaffna, ‘Chauakacheri Murunga, Chem, Kady, Palmurungai. Anonymous {1] lso reported Periyakulart 1 (PKM 1), ‘Until now a full characterisation of the oil produced from the seeds of Moringa oleifera Malawi using three different ways of extraction has not been reported. The oils were also com pared with virgin olive ol. MATERIALS AND METHODS Materials, ‘The seeds of Moringa oleifera wild local variety of Malawi (Blantyre area) were obtained from Malawi. The virgin olive "Horio" was purchased (rom Minerva S.A. (Athens, 14452, Greece}. Reagents All the reagents (analytical and HPLC grade) were obtained trom Sigma Chemicals Company Co. (St. Louis, MO 63178, USA) and the standard solutions for the determination of tocopherols were purchased by Merck Lid (Darmstand, D- 64271, Germany) (dl-a-tocophero!), Sigma ((+)-5-tocopherol), British Greyhound Chromatography and Allied Chemicals (Bi kenhead, Merseyside, L434X, UK)(Falty acid methylesters standards} and Laradan AB (Malm, $-21616, Sweden) (ster- standards). LUA AIVISTA TALIANA DELLE SOSTANZE GRASSE - VOL. LOKV - GENNAIO 1988 a1Oil extraction The seeds of Moringa oleitora were divided into three por tions and the oil was produced by cold pressure (CP) and ex tracted by tne use of hexane (H) and a mixture of chlor: form: methanol (50:50) (CM). as solvenis. “The extraction procedure for the cold pressure was per tormed as follows: the seeds wore milled to a fine paste with a Vorwerk Thermomix 3300 (Vorwerk France S.A., Patis) at 2 speed of 12 with the addition of water (ina ratio of 1 seedi2 wales prior to extraction which was done with a O.M.F-B. pra 25/1 simple hydraulic hand press (Costruz. Meco. Oteo- Ginamiche Provaglio D'seo, Brescia, italy) with a max. pres- sure of 300 kale. “The solvent extractions were executed using a two lite soxh- let apparatus (\e. “cold” solvent) Beore the beginning of the measurements the oil was re- fined (degummed) apart from a small quantity of about 40 mi which was kept for the induction time determination (Ranci mal) and other methods inorder to compare the unrefined with the refined ol, The solvent was evaporated under reduced pressure, and the oll rom diferent batches were combined ‘and kept in sealed bottles under refrigeration (0 ~4°C) for fur- ther processing and analysis, Degumming Tho oil was heated at 75 °C and 20% boiling water was ad ded. The mixture was mixed for 10 minutes with the aid of a glass f0d. After cooling, the oll wes centrifuged for 10 minutes in 3500 rpm in tubes of 200 cm? using a Sorvall General- Purpose RC-3 Automatic Refrigerated Centrituge Ivan Sor- vall Inc., Newtown, Connecticut 06470, USA). Determination of the physical characteristics ‘The determination of the physical characteristics was as fol- lows: the density (relative density 40 °C/20 °C), refractive in- dex (at 40 °C), color (measured with a Lovibond tintomeler) (The Tintometer Ltd., Salsbury, England) and smoke point ac- Ccotding to the method described by British Standards Methods ‘of Analysis, BS 684: Section 18) were measured. Determination of the chemical characteristics ‘The determination of the chemical characteristics was as follows: the acidity (measured according to the method described by IUPAC (13}, the saponification value (determined according to the method dascribed by AOCS method Cd 3-25, described in Bailey's Industrial Oil and Fat Products), the io- dine value (measured according to the Wijs method as described by Pearsons [22]. Determination of the fatty acid composition The determination of the fatty acids composition was done by gas-liquid chromatography according 10 the method described by Tsaknis [26] “The FAMES preparation was done using the following proce- dure: about 25 mg of ol were accurately weighed into a screw cap tube, and 1.8 cm? methanolic sodium hydroxide was ad- ded, mixed and heated at 100 °C for 7 minutes. After cooling, 2.cm? of boron trifluoride were added and heated al 100 °C. for 5 minutes. The tube was cooled to 30 - 40 °C and 1 om? Cf iso-octane was added, capped and shaken using whirti mix for 30 seconds. 5 cm? of saturated sodium chloride solution ‘was immediately added and the tube was shaken again. The tube contents were allowed to separate and the top (iso-octane containing faty acid methyl esters) layer was removed and the lower layer was extracted again with an addition of 1 crn? iso- ‘octane, The two iso-octane extracts were combined (dried over anhydrous sodium sulfate) and concentrated to approximate- ly tT em®with a stream of nitrogen, 22 Analysis of fatty acid methylesters was performed on a Var- ian 3600 Gas chromatograph (Varian, Palo Alto, California, USA) equipped with a Carbowax 20M (Supelco, Inc. Supeleo Park, Bellofonte, PA 16823-0048) 10°x1/8" (5% on Chromosor W 80/100 mesh) column. The temperature program was 60°C for 10 min and then 2°C min“' up to 220°C, Injector and FID temperatures were set at 160°C and 280°C respectively, sam ple volume was 0.2 yl, the carrier gas was No at a flow of 30 'm] min", chart speed was set at 0.5 cm mint and the attenu- ation at 10°1x32, In total three samples were prepared and measured separately. Determination of the sterol composition The identification and determination of sterols by GLC was done according to the methad described by the Official Jour- nal of the European community, No. L 248, 59.1991 ‘Analysis of sterols was performed on a Hewitt Packard 6890 Gas Chromatograph (Hewlett-Packard, San Diego, CA, USA) ‘equipped with a DB-S FSOT capillary column (80 mx 0.25 mm x 0.28 um) (J & W, 91 Blue Ravine rd., Folson, 95690-4714, California, USA), The pressure of the carrier gas (H2) was 75 pa. Injector and FIO temperatures were 280°C and 300°C. respectively. The temperature program was isothermal 260°C for 40 min at least, In total three samples were prepared and measured separately. Determination of the tocopherol composition ‘The method used for he determination ofocopherols was a modification ofthat reported by Carpenters 16} {@) 19 of all was accurately weighed info @ 3 gram sample Miatwappedin fol paper to prover oxidation, The of wesc solved in a § cm n-hexane before injection. (0) 20 ul campo was injected into the Waters 600 HPLC Pump (Miipore Corporation, Waters Chromatography Divison, Masgachasots, MA Y7S?, USA fitted wih @ Wars Plas, 425 h, 10 um, 39x200 mm column Bataaioh was portormad wth a Waters 486 Tunable Absor bance Detector sol at 298 nm. lsorpropanotn hevanerabso- Vue ethanol (:978:06) al 1 emb/min was sed o6 the moale phase, A toa of min was necessary o ascay tne tocopher- bls In total tee samples were prepared and moasured separately. Determination of the oxidative state For the determination of the oxidative state the peroxide value as well as the specitic extinction (E"S,) at 282. and 2370 nm were measured. The poroxide vallé was measured Using the method adapted trom Lea (1952). The determina. tion of specific extinction (E™, at 282 and 370 nm) was car- ried out using the method of {UPAC (1987) and by the use of, fa Hitachi U-3210 spectrophotometer (Hitachi Lid. Tokyo, Japan} Determination of the susceptibility 10 oxidation with the Ran- cimat method ‘Two and a half grams (2.5 g) of oll were accurately weighed into each of the six reaction vessels and the following proce- ‘dure was carried out. Tho “Metrohm Rancimat 679" (Metrohen LUd,, CH-3101, Herisau, Switzerland) was switched on until the temperature of the ol Batch reached the temperature of 120°C, ‘Then 50 cm9of distilled water was placed into each of the six conductivity cells and the air flow rate was set at 20 L t-!, The temperature was checked to ensure it had a constant value. ‘The air supply was connected to the tubes containing the oil samples and the chart recorder was started. The determina tion continued automatically until the conductivity reached the maximum value and the induction period was read. LA RIVISTA TALIANA DELLE SOSTANZE GRASSE - VOL. LXXV - GENNAIO 1908