反相高效液相色谱法测定血清中维生素E的含量

Determination of vitamin E in human serum by RP HPLC

目的：建立反相高效液相色谱（RPHPLA）测定人血清中维生素E（VE）浓度的方法。方法：移取血清，加入乙腈乙醇混合溶液及正己烷，旋涡混合、离心，分离正己烷层，氮气吹干，残渣用流动相溶解后进样。采用XTerra^Tm RP18色谱柱（5μm，3．9×150mm），流动相为甲醇：水（92：8，v／v）；流速1mE／min；柱温30℃；检测波长292nm。结果：VE的保留时间为7．61min，浓度在2-16μg／ml范围内线性关系良好（r^2=0．9999，n=5），平均回收率为95．6％，RSD〈6％，最低检测量2ng（S／N〉3）。40例健康成人血清VE平均值为10．20±2．51μg／ml。结论：反相高效液相色谱检测血清VE浓度的方法灵敏度高，回收率和重现性良好．操作快速、简便．这对测定血清中其它维生素含量具有指导意义。

Objective：To set up a method using high performance liquid chromatography （HPLC） to determine content of the vitamin E（VE） in human serum. Methods：The serum was extracted with n-hexane after the protein in the serum was deposited with acetonitrile/alcohol. The layer of n - hexane was dried with nitrogen, and the leavings dissolved with mobile phase and determined by HPLC. VE were separated on XTerra^TM RP18 5μm,3.9 × 150 mm column, and detected at 292 nm with UV detector. Using methanol/water（92：8 ,v/v） as mobile phase （current speed = 1 ml/min） when the column temperature was at 30℃. Results：The linearity range of calibration curve of VE was 2 - 16 μg/ml （ r^2 = 0. 9999 ）. The average extracted recovery was 95.6%, RSD 〈 6%. The detection limit of VE was 2 ng（ S/N 〉 3 ）. Average value of the serum VE of 40 healthy adults was （ 10. 20 ± 2. 51 ） μg/ml. Conclusion：The method is sensitive, rapid, accurate and simple. It would be of guiding significance to use the method to determine the content of other vitamins in human serum.