Separation of N-Methylhydroxamic Fatty Acids Based on Ketapang Seed Oil using High Performance Liquid Chromatography

Ana Restu Nirwana, D. Suhendra, E. Gunawan
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Abstract

N-methyl fatty hydroxamic acids (N-MFHAs) is a derivative of a hydroxamic acid (FHA), were synthesized from ketapang seed oil (Terminalia catappa L.) with N-methyl hydroxylamine enzymatically using immobilized lipase (Lipozyme TL IM). N-MFHAs that are synthesized are still in a mixed state of triglycerides of the ketapang seed oil. This research aims to determine the optimum conditions for the separation of N-MFHAs into their single components and their percentage composition using High Performance Liquid Chromatography (HPLC). The specification of the HPLC system includes using the reverse phase column SGE C-18 ODS-2 and UV detector 213 nm. The optimum conditions for the separation of N-MFHAs in the HPLC system include using the mobile phase of methanol 100% and flow rate of 0.25 mL/minutes with an injection volume of 20 µL and sample concentration of 10.000 ppm. The percentage of the N-MFHAs composition successfully separated using HPLC were linoleoyl-methyl-fatty hydroxamic acid (13.09%), oleoyl-methyl-fatty hydroxamic acid (62.46%), palmitoyl-methyl-fatty hydroxamic acid (19.51%), and stearoyl-methyl-fatty hydroxamic acid (4.93%).
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高效液相色谱法分离卡达蓬籽油中n -甲基羟肟脂肪酸
N-甲基脂肪异羟肟酸(N-MFHA)是一种异羟肟酸酯(FHA)的衍生物,以ketapang籽油(Terminalia catappa L.)为原料,利用固定化脂肪酶(Lipozyme TL IM)与N-甲基羟胺进行酶促合成。合成的N-MFHA仍然处于ketapang籽油的甘油三酯的混合状态。本研究旨在确定使用高效液相色谱法(HPLC)将N-MFHA分离为其单一组分及其百分比组成的最佳条件。HPLC系统的规格包括使用反相柱SGE C-18 ODS-2和UV检测器213nm。在HPLC系统中分离N-MFHA的最佳条件包括使用100%甲醇的流动相,流速为0.25 mL/分钟,进样体积为20µL,样品浓度为10.000 ppm。使用HPLC成功分离的N-MFHA组合物的百分比为油酰基-甲基脂肪异羟肟酸(13.09%)、油酰基-甲酯脂肪异羟酰胺酸(62.46%)、棕榈酰基-甲基油脂异羟酰胺(19.51%)和硬脂酰基-甲基脂酸异羟肟(4.93%)。
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