LC-MS/MS法测定人体血清中脂溶性维生素浓度  

Determination of fat-soluble vitamins in human serum by LC-MS/MS

作　　者：陈敏纯 张婧一 郑洁[1] 王新[1] 刘生元 曹青[1] 闫抗抗 Chen Minchun;Zhang Jingyi;Zheng Jie;Wang Xin;Liu Shengyuan;Cao Qing;Yan Kangkang(Department of Pharmacy,Xi’an No.3 Hospital,the Affiliated Hospital of Northwest University,Xi'an 710018,China)

机构地区：[1]西北大学附属医院/西安市第三医院药剂科,西安710018

出　　处：《分析仪器》2021年第6期135-139,共5页Analytical Instrumentation

摘　　要：目的:建立LC-MS/MS法同时测定人体血清中脂溶性维生素A、25-OH-D_(2)、25-OH-D_(3)、E的质量浓度。方法:以同位素标记的维生素A-d4、25-OH-D_(2)-d6、25-OH-D_(3)-d6、VE-d6作为内标,4%BSA溶液作为血清的替代基质溶液,正己烷进行萃取,质谱采用ESI正离子MRM模式,色谱柱选用Zorbax Extend-C_(18)柱(150mm×4.6mm,5μm,),流动相为0.1%甲酸水和0.1%甲酸甲醇,流速以0.6mL/min进行梯度洗脱。结果:维生素A、25-OH-D_(2)、25-OH-D_(3)、E的质量浓度在0.057~2.536μg/mL、0.004~0.200μg/mL、0.004~0.200μg/mL、0.409~20.480μg/mL范围内线性关系良好,其测定的日内和日间精密度RSD为0%~15%,回收率分别为95.04%~103.72%、93.00%~95.04%、105.99%~112.34%、101.32%~104.30%,基质效应分别为106.27%~109.07%、104.99%~108.47%、103.72%~108.15%、95.93%~99.20%。4%BSA替代基质溶液与血清基质溶液的一致性验证,检测均值和理论值相对误差<15%,变异系数<15%,均在可接受范围之内。结论:建立LC-MS/MS法测定人体血清中脂溶性维生素A、25-OH-D_(2)、25-OH-D_(3)、E的质量浓度具有精确度高,重现性好,特异性好,且线性关系良好,可满足高通量临床治疗药物浓度监测。Isotope labeled VA-d4,25-OH-D_(2)-d6,25-OH-D_(3)-d6,VE-d6 were used as internal standards,4%BSA solution was used as substitute for serum,the sample was extracted with N-hexane.The residuals were analyzed by LC-MS/MS system on Zorbax Extend-C_(18)column(150 mm×4.6 mm,5μm).The identification of the fat-soluble vitamins was performed under positive ions mode with multi-ion monitoring(MRM).The mobile phase was 0.1%formic acid-water(A)and 0.1%formic acid methanol(B)in gradient elution,and the flow rate was 0.6 mL/min.The linear range of the fat-soluble vitamin A,25-OH-D_(2),25-OH-D_(3),E was 50.057μg/mL—2.536μg/mL,0.004μg/mL—0.200μg/mL,0.004μg/mL—0.200μg/mL,0.409μg/mL—20.480μg/mL and the linear relationship was good.The intraday and interday precision were within 0%—15%.The recoveries were within 95.04%—103.72%,93.00%—95.04%,105.99%—112.34%,101.32%—104.30%,and the matrix effects were within 106.27%—109.07%,104.99%—108.47%,103.72%—108.15%,95.93%—99.20%,respectively.The consistency of 4%BSA solution and serum were verified.The comparison results showed that the discrepancy was within the acceptable range.The method is sensitive,accurate,reproducible and has good linear relationship,and is suitable for high-throughput analysis of therapeutic monitoring.

关 键 词：脂溶性维生素 定量检测 液质联用仪 

分 类 号：O657.63[理学—分析化学] R969[理学—化学]