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目的建立氨基酸自动分析仪测定保健食品中还原型谷胱甘肽的方法。方法样品经p H=2.20的盐酸-柠檬酸缓冲溶液溶解,超声提取及微孔滤膜过滤后,经p H=3.20及p H=4.25的盐酸-柠檬酸钠缓冲液梯度洗脱,采用阳离子交换树脂柱分离,还原型谷胱甘肽与茚三酮发生显色反应并在570 nm处波长下测定,柱温为44℃,流速为35 ml/h,反应温度为135℃,进样体积为20μl。结果 GSH对照品的浓度为1.0μg/ml~100.0μg/ml时,线性关系良好,线性回归方程为y=19.974x+12.379,相关系数(r)>0.999,定量限为0.63μg/ml,检出限为0.19μg/ml,平均加标回收率为102.1%,峰面积的相对标准偏差为1.8%。结论该方法简便、快速、灵敏、准确,适用于保健食品中还原型谷胱甘肽含量的测定。
Objective To establish a method of automatic amino acid analyzer determination of reduced glutathione in health food. Methods The sample was dissolved in hydrochloric acid-citric acid buffer solution with pH = 2.20. After ultrasonic extraction and microfiltration filtration, it was eluted with a gradient of hydrochloric acid-sodium citrate buffer of p H = 3.20 and p H = 4.25, Exchange resin column separation, reduced glutathione and ninhydrin color reaction and determination at 570 nm wavelength, the column temperature was 44 ℃, the flow rate of 35 ml / h, the reaction temperature was 135 ℃, the injection volume 20 μl. Results When the concentration of GSH reference substance was 1.0μg / ml ~ 100.0μg / ml, the linear regression equation was y = 19.974x + 12.379, the correlation coefficient (r)> 0.999, the limit of quantification was 0.63μg / ml The limit of detection was 0.19μg / ml. The average recovery was 102.1%. The relative standard deviation of peak area was 1.8%. Conclusion The method is simple, rapid, sensitive and accurate and suitable for the determination of reduced glutathione in health food.