脂肪種子が貯蔵するトリアシルグリセロール分子種のHPLCによる分析法の改良

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  • シボウ シュシ ガ チョゾウ スル トリアシルグリセロール ブンシシュ ノ HPLC ニ ヨル ブンセキホウ ノ カイリョウ

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Analysis of triacylglycerol (TAG) molecular species by high performance liquid chromatography was tested in terms of solvent constituents, column temperature and monitoring by UV. Tests were made on Dioscorea tokoro seed oil, which typically contains various TAG molecular species, and that of two other plants. When acetonitrile content was decreased from 700 60%, retention times of TAGs were reduced by half. Retention times did not change within 2-propanol and hexane ratios of one to five; however, for separation of partially overlapped peaks, 2-propanol/hexane=2/1 was the most suitable ratio. When column temperature was lowered from 37℃ to 19℃, retention times lengthened two to three times depending upon TAG molecular species. Based on these results, we propose a solvent composition of acetonitrile/2-propanol/hexane=6/2/1 (v/v) and column temperature of 24℃ for clear separation of TAG molecular species. As the number of double bonds increased, absorption intensity at 210nm increased; TAG molecular species without a double bond were also detectable at 210nm. Because relations of dose-peak area of TAGs were shown by straight lines for the wide concentration range, monitoring at 210nm enables quantification of TAG molecular species. Detection limits ranged from 0.02μg (trilinolenin: total number of double bonds was nine) to 2.0μg (tripalmitin: no double bond). The method introduced here clearly separated and sensitively detected Pterocarya stenoptera seed oil, which contains many TAG molecular species of unsaturated fatty acids.

source:The technical bulletin of Faculty of Horticulture, Chiba University

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