超高效液相色谱法测定人血清中维生素C含量

Determination of vitamin C in human serum by ultra-performance liquid chromatography

目的建立超高效液相色谱法测定人血清中维生素C含量的方法。方法血清经过偏磷酸沉淀蛋白后,以3,4-二羟基苄胺氢溴酸盐为内标,使用HSS T3色谱柱(2.1 mm×100 mm,1.8μm)分离,以20 mmol/L磷酸二氢钾水溶液和甲醇作为流动相,等度洗脱分离。采用内标法定量。结果维生素C能在5 min内分离,在0.287~28.7μg/mL浓度范围内线性关系良好。方法的检出限为0.0209μg/mL,定量限为0.0697μg/mL。加标回收率为88.4%~108.4%,相对标准偏差为1.76%~4.92%(n=6)。结论该方法准确、灵敏度高、前处理简单,能够检测血清中的维生素C。

OBJECTIVE To establish a method for the determination of vitamin C(ascorbic acid)in human serum by ultra-performance liquid chromatography(UPLC).METHODS The sample was deproteinized by metaphosphoric acid,added 3,4-dihydroxybenzylamine hydrobromide as internal standard.The analysis was performed on HSS T3 column(2.1 mm×100 mm,1.8μm),and isometric elution using 20 mmol/L KH;PO;aqueous solution and methanol as the mobile phase.The samples were quantified by the internal standard method.RESULTS Vitamin C was separated within 5 minutes.The linear was good for Ascorbic acid in the range of 0.287-28.7μg/mL.The limits of detection for vitamin C was 0.0209μg/mL.The limit of quantitation was 0.0697μg/mL.Recoveries of vitamin C were in range of 88.4%-108.4%,with the relative standard deviations were 1.76%-4.92%(n=6).CONCLUSION This method is fast,accurate and sensitive and the preprocessing is simple,which can be used for determination of vitamin C in human serum effectively.