Dertermination of reduced glutathione (GSH) and thiols in proteins (P-SH) in 4 kinds of nuts by high performance liquid chromatography (HPLC)
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摘要: 用5,5’-二硫双-2-硝基苯甲酸（DTNB）为柱前衍生化试剂,采用细胞破壁、衍生化为一体的同步衍生化提取,HPLC梯度洗脱,测定了市售开心果、巴旦木、澳洲坚果、腰果干品中谷胱甘肽（GSH）和含半胱氨酸残基的蛋白巯基（P-SH）含量。结果发现四种坚果均含较丰富的GSH和P-SH巯基,其含量分别为,开心果:GSH（1.2303±0.01126）μmol/g,P-SH（3.5414±0.0531）μmol/g;巴旦木:GSH（0.3069±0.0185）μmol/g,P-SH（1.9001±0.0194）μmol/g;澳洲坚果:GSH（1.5708±0.0216）μmol/g,P-SH（0.5905±0.0238）μmol/g;腰果:GSH（0.3373±0.0047）μmol/g,P-SH（2.2458±0.0225）μmol/g。在优化的选定条件下,测定方法的回收率为99.37%101.45%,GSH和P-SH测定检测限分别为0.1386μmol/g和0.2184μmol/g。本研究对坚果活性成分的构成及其巯基化合物在保健防病作用研究及其功能性食品开发均提供了有用的参考数据。Abstract: The contents of reduced glutathione ( GSH) and protein thiols in prunus duleis, cashew nut, pistachio nut, macadamia nut were detected by pre-column derivatization HPLC method using 5, 5'-dithio-bis-2-nitrobenzoic acid ( DTNB) as derivatizing agent with concomitant cell rupture sample pretreatments.The results showed that detected nuts contained relatively abundant GSH and thiols in P-SH. The amounts detected were GSH ( 1.2303 ± 0.01126) μmol/g, P-SH ( 3.5414 ±0.0531) μmol/g for pistachio nut, GSH ( 0.3069 ± 0.0185) μmol/g, P-SH ( 1.9001 ± 0.0194) μmol/g for prenus duleis, GSH ( 1.5708 ± 0.0216) μmol/g, P-SH ( 0.5905 ± 0.0238) μmol/g for macadamia nut, GSH ( 0.3373 ± 0.0047) μmol/g, P-SH ( 2.2458 ± 0.0225) μmol/g for cashew nut respectively.The recoveries of the methods were 99.37% 101.45%, and detection limits were 0.1386 μmol/g for GSH and 0.2184 μmol/g for protein thiols.The results provided a useful data for the application of these nuts for the healthcare.
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Keywords:
- HPLC /
- nuts /
- glutathione /
- thiol compounds
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