UPLC法测定儿童血浆中维生素A

目的：临床上建立一种快速、准确测定儿童血浆中维生素A浓度方法。方法：取儿童血浆200μl，加入140μl甲醇和异丙醇（v／v，8：2），涡旋30s后，加入1000μl正已烷，涡旋1min，室温高速离心2min（12000×g），取上清液700μl，吹干后甲醇100μl复溶。色谱条件：色谱柱为13EHC18柱（50mm×2．1mm，1．7μm；Waters公司）；流速为0．3ml·min^-1；流动相为甲醇-水（90：10）；柱温为30℃；进样量：5乩；检测波长为325nm。结果：血浆中维生素A色谱峰分离良好，无干扰，维生素A曲线为Y=118296x＋11644，R2=0．9997，回收率在90—120％。结论：本法首次使用UPLC法测定血浆中维生素A，且操作简便，适用于儿童血浆中维生素A快速监测。

The determination of Vitamin A in human plasma by UPLC

Objective To establish a UPLC assay to determine the concentration of Vitamin A in human plasma for clinical treatment of a patient. Methods 140 μL of methanol -2 - propanol （80：20 by volume） were added to 0. 2 mL of Children＇ s serum. The tubes were mixed in a Muhitube vortex mixer for 30 s. Vitamin A was extracted by mixing one times （60 s） with 1 mL of hexane. The phases were separated by centrifugation, and the upper organic phase was transferred to a conical tube and dried under nitrogen. The residue was dissolved in 5μL of methanol. Vitamin A was successfully separated on the BEH C18 （50 mm×2. 1 mm, 1.7μm; Waters Corporation） column at a flow -rate of 0. 30 mL·min-1, with the mobile phases of methanol -water. The column temperature was set at 30℃. The wavelength for detected was 254nm. Results The linear equation was y = 118296x ＋ 11644, R2 = 0. 9997, R2 = 0. 9990. Conclusion The method is convenient and suitable for the determination of Vitamin A in Children＇ s plasma.