液相色谱串联质谱法测定血清维生素A、E含量

目的 建立液相色谱串联质谱法(LC-MS/MS法)测定血清维生素A和维生素E含量的方法.方法 采用硫酸锌沉淀蛋白,96孔蛋白沉淀板过滤.采用LCMS/MS正离子电喷雾离子化(ESI+)的多反应监测模式(MRM)]氘代同位素内标法检测血清维生素A、E含量并进行相关方法学验证.结果 LC-MS/MS检测血清维生素A的批内精密度变异系数为3.2％(2.4％～3.9％),批间变异为5.5％(5.O％～6.0％);维生素E的批内精密度变异系数为4.1％(3.0％～5.1％),批间变异为8.0％(7.6％～8.3％).维生素A在0.06 ～2 μg/mL、维生素E在1.2 ～ 40μg/mL范围内线性良好,线性相关系数分别为0.9987、0.9909.维生素A、维生素E的定量限分别为0.03μg/mL、0.8μg/mL;检测限分别为0.02μg/mL and 0.01 μg/mL.维生素A低浓度回收率为107.5％,高浓度为106.9％;维生素E的低浓度回收率为91.9％,高浓度为88.4％.与液相色谱方法进行比对,两种方法的结果差异无统计学意义(P值均大于0.30)说明两种方法具有一致性.结论 LC-MS/MS检测血清维生素A及维生素E的灵敏度高,结果准确、稳定,可应用于临床分析.

Determination of Vitamin A and E in Serum by Liquid Chromatography Tandem-mass Spectrometry

Objective To establish a method for the determinations of serum vitamin A and vitamin E contents by liquid chromatography tandem-mass spectrometry (LC-MS/MS).Methods After serum levels of proteins were precipitated by zinc sulfate and filtrated by 96 pore protein-precipitate plate,then analyzed by LC-MS /MS in the positive electrospray ionization (ESI +) mode and multiple reaction monitor (MRM) mode.The quantitative analysis for vitamin A and vitamin E contents was carried out by deuterium isotope as internal standard,and the methodology validation was performed.Results The within-run precision of vitamin A was 3.2％ (2.4％-3.9％),and between-run precision of vitamin A was 5.5％ (5.0％-6.0％).The withinrun precision of vitamin E was 4.1％ (3.0％-5.1％),and between-run precision of vitamin E was 8.0％ (7.6％-8.3％).The good correlation coefficients were 0.9987 and 0.9909 at the concentration of 0.06-2pg/mL vitamin A and 1.2-40μg/mL vitamin E,respectively.The detection limits of vitamin A and vitamin E were 0.03g/mL and 0.8g/mL respectively.The quantitation limits of vitamin A and vitamin E were 0.02ug/mL and 0.01 ug/mL respectively.Low and high concentration of vitamin A recovery rate were 107.5％ and 106.9％ respectively;low and high concentration of vitamin E recovery rate were 91.9％ and 88.4％ respectively.Compared with the liquid chromatography method,the results of the two methods were not statistically significant(P ＞ 0.30),indicating the consistency of the two methods.Conclusion This method is sensitive,accurate and stable,satisfying the quantification requirements of vitamin A and vitamin E in serum.So it can be used in clinical analysis.