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目的:建立用高效液相色谱法测定维生素E烟酸酯血药浓度的方法。方法:用乙腈沉淀蛋白,正己烷提取血浆中的维生素E烟酸酯,离心后取上清液,氮气吹干,甲醇定量溶解后,进行高效液相色谱测定。色谱柱为DiamonsilODS(4.6mm×150mm,5μm)柱,流动相为甲醇,流速为1.5mL·min-1,柱温35℃,检测波长为220nm。结果:该方法在0.109～2.725μg.mL-1浓度范围内线性关系良好(r=0.9991,n=6)。高、中、低3种浓度的日内、日间RSD均小于5%,血样5d的稳定性RSD也小于5%,方法回收率在90.6%～99.9%之间。结论:本方法简便、定量准确,适用于人血浆中维生素E烟酸酯的测定,以及维生素E烟酸酯人体药代动力学、生物利用度和生物等效性的研究。
Objective: To establish a method for the determination of vitamin E nicotinate concentration in plasma by high performance liquid chromatography. Methods: The protein was precipitated with acetonitrile and n-hexane was used to extract the vitamin E nicotinate in plasma. After centrifugation, the supernatant was taken out and dried with nitrogen gas. After the methanol was dissolved quantitatively, the content of vitamin E was measured by HPLC. The column was Diamonsil ODS (4.6 mm × 150 mm, 5 μm) with a mobile phase of methanol at a flow rate of 1.5 mL · min -1 and a column temperature of 35 ° C. The detection wavelength was 220 nm. Results: The method showed good linearity (r = 0.9991, n = 6) in the range of 0.109 ~ 2.725μg.mL-1. The RSDs of intra-day and inter-day high, medium and low concentrations were all less than 5%. The stability RSD of blood samples for 5 days was less than 5%, and the recovery rates were between 90.6% and 99.9%. Conclusion: The method is simple and accurate. It is suitable for the determination of vitamin E nicotinate in human plasma and the human pharmacokinetics, bioavailability and bioequivalence of vitamin E nicotinate.