Chen Xi, Gong Zhaolong, Shen Shi
Key Laboratory of Trace Element Nutrition, National Health Commission of the People's Republic of China, National Institute for Nutrition and Health, Chinese Center for Disease Control and Prevention, Beijing 100050, China.
Wei Sheng Yan Jiu. 2021 Mar;50(2):301-307. doi: 10.19813/j.cnki.weishengyanjiu.2021.02.023.
To establish a method for the determination of vitamin A(retinol) and four active forms of vitamin E(α-tocopherol, β-tocopherol, γ-tocopherol and δ-tocopherol) in human serum by ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry(UPLC-MS/MS).
The sample was deproteinized by methanol, then extracted by n-hexane, dryness under nitrogen and followed by a reconstitution with methanol. The analysis was performed on a C_(30) column(3 mm×150 mm, 2. 6 μm), and isometric elution using 0. 1% formic acid in methanol and 5 mmol/L ammonium formate in 0. 1% formic acid as the mobile phase. The samples were determined by mass spectrometry in the positive ion mode with the multiple reaction monitoring mode, quantified by the internal standard method.
Vitamin A and four active forms of vitamin E were separated within 42 minutes, and β-tocopherol and γ-tocopherol can be distinguished. The linear was good for retinol in the range of 0. 0050-2. 5 μg/mL, 0. 036-20 μg/mL for α-tocopherol, 0. 042-8. 0 μg/mL for β-tocopherol and 0. 020-10 μg/mL for the other tocopherols. The limits of detection for retinol and tocopherols were in the range of 5. 76-31. 6 ng/mL. Recoveries of retinol and tocopherols at different levels were in range of 84. 4%-118. 6%, with the relative standard deviations were 1. 22%-8. 50%(n=6).
This method is fast, accurate and sensitive and the preprocessing is simple, which can be used for determination of vitamin A and four active forms of vitamin E in human serum effectively.
建立超高效液相色谱-串联三重四极杆质谱法（UPLC-MS/MS）测定人血清中维生素A（视黄醇）及维生素E的4种活性形式（α-生育酚、β-生育酚、γ-生育酚和δ-生育酚）的方法。
样品用甲醇进行蛋白沉淀，然后用正己烷萃取，氮气吹干后再用甲醇复溶。采用C30柱（3 mm×150 mm，2.6 μm）进行分析，以甲醇中0.1%甲酸溶液和0.1%甲酸溶液中5 mmol/L甲酸铵溶液作为流动相进行等度洗脱。采用正离子模式、多反应监测模式进行质谱测定，内标法定量。
维生素A及维生素E的4种活性形式在42分钟内实现分离，β-生育酚和γ-生育酚可区分。视黄醇在0.00502.5 μg/mL范围内线性良好，α-生育酚在0.03620 μg/mL范围内线性良好，β-生育酚在0.0428.0 μg/mL范围内线性良好，其他生育酚在0.02010 μg/mL范围内线性良好。视黄醇和生育酚的检测限在5.7631.6 ng/mL范围内。不同水平视黄醇和生育酚的回收率在84.4%118.6%之间，相对标准偏差为1.22%~8.50%（n = 6）。
该方法快速、准确、灵敏，预处理简单，可有效用于人血清中维生素A及维生素E的4种活性形式的测定。
