摘要
目的:建立超高效液相色谱-串联质谱(UPLC-MS/MS)同时测定血清中维生素A、E的方法。方法:取标准系列工作溶液、空白替代血清样品(4%BSA牛血清白蛋白溶液)各50μL,经硫酸锌、沉淀剂(甲醇/乙腈=50/50,V/V)沉淀蛋白,振荡,12000 r/min离心5 min,取上清,低压离心挥干,复溶振荡,12000 r/min离心5 min,取上清待测。采用Waters BEH Phenyl色谱(2.1 mm×100 mm,1.7μm)分离,以0.1%甲酸-水溶液和0.1%甲酸-甲醇溶液为流动相等度洗脱,电喷雾(ESI)正离子模式、多反应监测模式(MRM)下检测,内标法定量。结果:UPLC-MS/MS检测血清维生素A、α-维生素E、β-维生素E线性关系良好,相关系数分别为0.9993、0.9993、0.9997;维生素A、α-维生素E、β-维生素E定量限分别为0.213、0.240、0.070 ng/mL,检出限分别为0.064、0.072、0.021 ng/mL;维生素A、E在低、高浓度加标回收率范围分别为89.7~107.4、97.6~118.1,RSD分别为2.45%~3.43%、1.48%~5.40%。结论:本研究建立的人血清维生素A、E超高效液相色谱-串联质谱法灵敏度高、准确稳定、重现性好,对血清中维生素A和维生素E的测定具很好的的应用价值。
Objective To establish a method for simultaneous determination of serum vitamin A and E by ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method.Method Take 50μL each of standard series working solution and blank replacement serum sample(4%BSA),after proteins were precipitated by zinc sulfate and precipitant(methanol/acetonitrile=50/50,V/V),take the supernatant for testing after pre-processing,and then separated by Waters BEH Phenyl(2.1mm×100 mm,1.7μm)and eluted with the mobile phaes consisted of 0.1%formic acid in water and methanol in a equals program,then analyzed by UPLC-MS/MS with the positive electrospray ionization(ESI+)mode and multiple reaction monitor(MRM)mode,internal standard method for quantification.Result UPLC-MS/MS detection of serum vitamin A,α-vitamin E,β-vitamin E had a good linear relationship,the correlation coefficients were 0.9993,0.9993 and 0.9997,respectively.The quantitation limits of vitamin A,α-vitamin E,β-vitamin E were 0.213 ng/mL,0.240 ng/mL and 0.070 ng/mL,respectively,the detection limits of vitamin A,α-vitamin E,β-vitamin E were 0.064 ng/mL,0.072 ng/mL and 0.021 ng/mL.Low and high concentration of vitamin A and vitamin E recovery rate were 89.7~107.4 and 97.6~118.1 respectively,RSD were 45%~3.43%and 1.48%~5.40%.Conclusion This proposed method is sensitive,accurate and stable,reproducible,and has very good application value for the determination of vitamin A and vitamin E in serum.
作者
王睿
付志成
胡贻椿
毛德倩
杨晓光
陈竞
杨丽琛
WANG Rui;FU Zhi-cheng;HU Yi-chun;MAO De-qian;YANG Xiao-guang;CHEN Jing;YANG Li-chen(National Institute for Nutrition and Health,Chinese Center for Disease Control and Prevention/Key Laboratory of Trace Element Nutrition,National Health Commission of the People s Republic of China,Beijing 100050,China)
出处
《中国食物与营养》
2020年第9期40-44,共5页
Food and Nutrition in China
