RP-HPLC同时测定人血清中维生素A和维生素E

Simultaneous determination of retinol and α tocopherol in human serum by RP HPLC

目的：测定不同病理生理状态下人血清中维生素Ａ、维生素Ｅ的浓度，寻求弄清与某些疾病的关系，为临床治疗学、营养学等研究提供参考。方法：取血清２００μｌ加入含内标的无水乙醇液２００μｌ，振荡，加入正己烷５００μｌ，离心，取正己烷层１００μｌ进样；色谱条件为：色谱柱ＷａｔｅｒｓμＢｏｎｄａｐａｋＣ１８柱（３．９ｍｍ×３０ｃｍ），流动相甲醇水（９６∶４），内标维生素Ａ醋酸酯，检测波长３３０ｎｍ，２９２ｎｍ。结果：维生素Ａ在８６．８０４８～９７２．２１３０μｇ·Ｌ－１浓度范围内呈线性，ｒ＝０．９９９５，维生素Ｅ在１．２６６５～２５．３３１２ｍｇ·Ｌ－１浓度范围内呈线性关系，ｒ＝０．９９９６；维生素Ａ和维生素Ｅ的平均回收率分别为９６．４０％（ＲＳＤ＝３．９８％）和１０３．６７％（ＲＳＤ＝４．２６％），测定了２１８例人血清中维生素Ａ、维生素Ｅ的浓度。结论：方法快速、灵敏、准确，样品处理简便易行。

OBJECTIVE:To determine the concentrations of retinol and α tocopherol in human serum.METHODS:To 200 μl serum was added 200 μl anhydrous alcohol containing internal standard in a vessel.After 500 μl of n hexane was added,the mixture was shaken and centrifuged.The organic layer 100 μl was injected.The HPLC system consisted of μ Bondapak C 18 column,mobile phase of methanol:water(96∶4) mixture,internal standard of retinyl acetate,UV detector at 330 nm(retinol) and 292 nm(α tocopherol).RESULTS:The linear ranges of determination of retinol and α tocopherol were 86.804 8～972.213 0 μg·L -1 (r=0.999 5) and 1.266 5～25.331 2 mg·L -1 (r=0.999 6),respectively.The mean recovery of 96.96%(RSD=3.78%) for retinol and 102.03 %(RSD=3.93%) for α tocopherol was obtained.Blood from 218 human volunteers was collected and retinol and α tocopherol levels in serum were determined with this method.CONCLUSION:The method is sensitive,rapid and accurate.