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Note Fatty and Amino Acid Compositions of Vernicia Fordii Indian Journal of Experimental Biology Vol. 55, October 2017, pp. 734-739 Note Fatty and amino acid compositions of Argentina and Paraguay are the countries where it spread widely followed by Africa, India as well as Vernicia fordii: a source of 3 α-eleostearic acid and methionine southern regions of United States . In 2012, China produced about 80% of the world production of tung 4 Sandeep Kumar1*, MK Dhillon2, M Singh3, RS Rathi4, AK oil . The word ‘Tung’ is an ancient Chinese term for Misra5 & JC Rana1 heart. Today, the term is also used to refer the heart 1 1Division of Germplasm Evaluation; 4Division of Plant shaped, large and dark green leaves of this tree . Exploration and Germplasm Collection, Tung plant is of great economic importance due to its ICAR-National Bureau of Plant Genetic Resources, novel high value oil. Tung oil, derived from seeds, is non- New Delhi-110 012, India edible and classifies as a conjugated polyunsaturated 2 Division of Entomology, ICAR-Indian Agricultural Research drying oil as it contains high content of cis, trans, trans- Institute, New Delhi-110 012, India 9, 11, 13-octa-decatrienoic acid5. It is commonly used in 3ICAR-National Bureau of Plant Genetic Resources Regional Station, Shimla-171 004, India formulations of coatings, resins, dyes and inks due to its 6,7 5ICAR-National Bureau of Plant Genetic Resources unique property to dry to a clear and hard finish . Dried Regional Station, Shillong-793 103, India tung oil does not allow many chemicals to pass through Received 05 September 2016; revised 29 November 2016 and also remains unaffected by environmental conditions such as moisture and temperature. Tung oil, unlike other Tung seed [Vernicia fordii (Hemsl.) Airy Shaw, Fam. drying oils, does not darken with the passage of time. It is Euphorbiaceae] was analysed for α-eleostearic acid content along because of these properties that tung oil makes a widely with seed meal amino acid composition to determine its industrial used drying ingredient in paints, varnishes, coatings and applications. Tung seed contains high contents of oil and protein 8,9 with 54.4 and 18.55%, respectively. Saturated, monounsaturated finishes . Tung oil has been explored as a raw material 10-12 and polyunsaturated fatty acids accounted for 4.98, 6.50 and to produce biodiesel , polyurethane and wood flour 88.51%, respectively. α-Eleostearic acid was the major fatty acid composites13, thermosetting polymer14 and repairing with 81.02%, along with substantial amounts of oleic (6.50%) and agent for self-healing epoxy coatings15. It is also used in linoleic (7.49%) acids. Similarly, tung defatted seed meal was found rich in sulphur containing amino acid i.e., methionine large scale for production of linoleum, oil cloth, brake (614.59 mg/100 g), and arginine (244.64 mg/100 g). The higher linings, soap, leather dressings, inks, insulating concentration of α-eleostearic acid makes tung seed oil suitable for compounds and fibreboard. Its oil cake is a good fertilizer use as drying ingredient in paints, varnishes and coatings. The but is unsuitable as an animal feed1. But after extracted amino acids/proteins or detoxified tung seed meal can be detoxification, it can be used as a feed ingredient for used as nutritional additive in feedstuffs. Further, the determined cetane index, iodine value and saponification number were not in cattle. accordance with the European Standard Organization. However, Despite some studies carried out on tung in China, it these biodiesel production related properties can be improved after is still an under-utilized plant. No such studies have mixing with other oils. been conducted in India. Earlier studies although Keywords: Aleurites fordii, Biodiesel, Drying ingredient, Mizoram, reported fatty acid composition, but their results are 4,16 Nutritional additive, Shillong, Sulphur containing amino highly variable . Information available on amino acid acids, Tung seed oil extraction from tung seed meal as an additive in feedstuffs is also scarce17. Therefore, present study was Vernicia fordii (Hemsl.) Airy Shaw (previously classified undertaken to determine the fatty acid and amino acid as Aleurites fordii) belongs to the family Euphorbiaceae compositions in tung seeds collected from northeastern and its common names are tung tree, tung oil tree, tung part of India. nut tree, China wood tree, China wood oil tree, Aceite 1 Materials and Methods chino, Lumbang oil, etc. Originally, it was reported in southeast Asian region, but later on, it was also grown Seed material in Malawi, Argentina and the USA2. Western-China, The seed material of tung germplasm IC278136 was initially collected from Champhai (Mizoram), India. ————— *Correspondence: The seedlings were planted in 2003 in the field gene E-mail: [email protected] bank of National Bureau of Plant Genetic Resources KUMAR et al.: FATTY AND AMINO ACID COMPOSITIONS OF VERNICIA FORDII 735 Regional Station, Shillong. The plant takes 6-7 years for maturity and yields about 4-5 kg fruits per annum. The tree reaches maximum productivity at around 10-12 years of age. The biochemical analysis of the seeds received from field genebank, ICAR-NBPGR Shillong station, was carried out at Division of Germplasm Evaluation, ICAR-NBPGR, New Delhi. The material is also conserved at National Genebank, ICAR-NBPGR, New Delhi. Fig. 1 depicts the seed (A) and its kernel (B). Fig. 1 — Seed (A); and kernel (B) of tung (Vernicia fordii) tree. Oil, protein and moisture content Oil content of seeds was determined by petroleum reagent kit (WAT052880-Waters Corporation, USA). ether extraction in a soxhlet apparatus for 16 h The vials were heated for 10 min in a water bath at according to the AOAC procedure 7.04818. Protein 55C before analysis with high performance liquid content was estimated by conventional Micro-Kjeldahl chromatography (HPLC). The AccQ-Fluor amino acid method 984.13 of AOAC19. Moisture content was derivatives were separated on a Waters 2707 Module determined by oven dried method. HPLC System attached to a PDA 2998 model. The Fatty acid analysis using gas liquid chromatography column temperature was set at 37C. The amino acids Seed kernels were grounded and 100 mg test sample were detected at 254 nm. The peaks were acquired was extracted with 10 mL of the extraction solution using Empower Pro Software® by Waters Corporation consisting of chloroform: hexane:methanol (8:5:2 v/v/v). (2005-08) and quantification was done based on The extracts were evaporated in nitrogen gas at 60C for standard amino acids calibration (Thermo Scientific 30 min. Fatty acids were converted to respective methyl Amino Acid Standard H, Prod # NCI0180) run at five esters as per method of Neff et al.20. One µL of the methyl concentrations viz., 10, 20, 30, 40 and ester solution was injected into a highly polar HP 50 µL. Amino acid peaks were visually checked to Innowax capillary column (30 m length, 0.32 m diameter, verify the results. The proportional molar 0.5 µm thickness) with a split ratio of 1:80. A Hewlett concentration for each amino acid was calculated based Packard gas chromatograph equipped with flame on the concentration of standard amino acids and ionization detector (model 6890) was used for the expressed as mg amino acid/100 g sample on dry analysis. Injector and detector temperatures were 260 and weight basis. 275C, respectively. The temperature of the oven was Statistical analysis programmed from 150C holding at 1 min to 210C at the Fatty acid composition was analysed in three rate of 15C/min, followed by 210 to 250 ⁰C at the rate replications whereas all other observations were taken of 5C/min for 12 min. Peaks were identified by in two replications. The data was analysed using comparing their retention time with that of standards. Microsoft Excel 200724. All data is expressed as Peak integration was performed by applying HP3398A mean±Standard error of replicates. software. Results Iodine value (IV), Saponification number (SN) and Cetane index (CI) Oil content and Fatty acid composition Iodine value, saponification number and cetane Seed oil, protein and moisture contents obtained index of tung oil were calculated empirically from its were 54.4, 18.55 and 10.01%, respectively. The oil fatty acid constituents21,22. quality is determined by its fatty acid composition. Amino acid analysis Analysis of tung oil composition showed the presence The 20 mg seed was lyophilized and the vacuum- of palmitic acid (2.39±0.04%), stearic acid dried samples were hydrolyzed as described by Dhillon (2.59±0.05%), oleic acid (6.50±0.08%), linoleic acid et al.23. The samples were dried in an oven at 114 ± 2C (7.49±0.09), and α-eleostearic acid (81.02±0.19%). for 24 h. After hydrolysis, excess HCl was removed Fig. 2 shows the different fatty acids separated using and the samples were vacuum dried for gas chromatograph along with respective retention 90 min. The seed samples were reconstituted with times. α-Eleostearic acid (18:3 Δ9cis,11trans,13trans) was 200 µL of 20 mM boiling HCl. The reconstituted found to be the major fatty acid accounting for 81.02% 20 µL samples were derivatized with AccQ-Fluor of the total fatty acids. Tung seed oil contains 4.98% 736 INDIAN J EXP BIOL, OCTOBER 2017 Fig. 2 — Chromatogram showing separated fatty acids of tung seed oil using GC. saturated fatty acids as palmitic and stearic fatty acids, Table 1— Amino acid composition of defatted tung seed meal 6.50% monounsaturated fat as oleic acid, and 88.51% polyunsaturated fatty acids as linoleic and Amino acids Tung cake (mg/100g) octadecatrienoic acid (α-eleostearic acid). The SFA: Asp 68.39 ±0.01 MUFA:PUFA ratio of oil was 1:1.30:17.77.
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