高效液相色谱法测定血清中维生素A的含量

Determination of Vitamin A in Serum by HPLC.

目的探讨高效液相色谱法(high performance liquid chromatography,HPLC)测定血清中维生素A含量。方法取血清200μL,用无水乙醇沉淀蛋白,再加入正己烷震荡提取维生素A,取一定量烷层经氮吹挥干,残渣加无水乙醇溶解后采用HPLC法分析,保留时间定性,标准曲线法定量。分析柱为C18柱(250 mm×46 mm,5μm),流动相:甲醇+水(98+2),检测波长325 nm。结果维生素A在0.25～125.50μg/mL之间线性良好(r>0.999 6),平均回收率为98.20%,RSD为0.85%。结论 HPLC法测定血清中维生素A的含量简便、快速,灵敏度高,准确性好。

Objective The feasibility that determination of Vitamin A in serum by HPLC（High Performance Liquid Chromatography） was investigated.Methods Taking 200 μL serum and the protein was precipitated with ethanol.Then n-hexane was added and shocked to extract Vitamin A.A certain amount of n-hexane layer evaporated by nitrogen-blow.The residue that dissolved by ethanol was analyzed by HPLC.The analytes were qualitated by retention time and quantitated by standard curve method.The analysis column of this method was C18 column（250mm×46mm,5 μm）,the flow phase was methanol∶CH3-OH＋H2O（98＋2） and detection wavelength was 325nm.Results The calibration plot was found to be linear in the range of 0.25～125.5 μg/ml for Vitamin A（r0.999 6）.The average recovery was 98.2 % and RSD was 0.85 %.Conclusion The method that determination of Vitamin A in serum by HPLC was simple,sensitive,rapid and accurate.