论文部分内容阅读
目的建立快速测定人血浆和全血中还原型谷胱甘肽浓度的液相色谱-串联质谱法。方法以沉淀法快速处理全血及血浆样品,采用阿昔洛韦为内标,SymmetryShieldTM RP18色谱柱,流动相为乙腈-0.1%甲酸水溶液(6∶94,V∶V),流速为0.3 mL·min-1,进样量为10μL;采用电喷雾(ESI)离子源,正离子电离模式,多反应离子监测(MRM):还原型谷胱甘肽,[M+H]+m/z 308.2→179.1;阿昔洛韦,[M+H]+m/z 226.2→152.0。结果还原型谷胱甘肽血浆浓度线性范围为0.050～2.00 mg·L-1和全血浓度线性范围为50.0～500 mg·L-1,血浆和全血中的最低定量限分别为0.050 mg·L-1和50.0 mg·L-1;提取平均回收率在90%以上,批内、批间精密度均小于15%,准确度均在85%～115%范围内。采集的全血样品在4℃可保持1 h稳定,样品处理后在-70℃可保存1 wk左右待测。结论本实验建立的方法操作简单快速、灵敏度高、准确度好,临床应用方便,可用于人血浆和全血中还原型谷胱甘肽浓度的测定。
OBJECTIVE To establish a liquid chromatography-tandem mass spectrometry method for rapid determination of reduced glutathione in human plasma and whole blood. Methods The whole blood and plasma samples were rapidly processed by precipitation method using acyclovir as internal standard. SymmetryShieldTM RP18 column was used with the mobile phase consisted of acetonitrile-0.1% formic acid aqueous solution (6:94, V:V) and the flow rate was 0.3 mL · min-1, injection volume 10 μL; electrospray (ESI) ion source, positive ionization mode, MRM: reduced glutathione, [M + H] + m / z 308.2 → 179.1; Acyclovir, [M + H] + m / z 226.2 → 152.0. Results The linear range of plasma concentration of reduced glutathione was 0.050-2.00 mg · L-1 and the linear range of whole blood concentration was 50.0-500 mg · L-1. The lowest limit of quantification in plasma and whole blood was 0.050 mg · L-1 and 50.0 mg · L-1. The average recoveries were over 90%. The intra- and inter-batch precision was less than 15% and the accuracy was within the range of 85% -115%. The collected whole blood samples can be kept stable at 4 ℃ for 1 h, and can be stored at -70 ℃ for about 1 wk after the sample is processed. Conclusion The method established in this experiment is simple, rapid, sensitive, accurate and easy to use. It can be used to determine the concentration of reduced glutathione in human plasma and whole blood.