高效液相色谱串联质谱法同时测定人血清8种叶酸循环代谢物  

Simultaneous detection of 8 folate metabolites in human serum using LC-MS/MS

作　　者：李慧敏 刁嘉茵 郑康帝 张召 林炳耀 林东子 LI Hui-min;DIAO Jia-yin;ZHENG Kang-di;ZHANG Zhao;LIN Bin-yao;LIN Dong-zi(Dongguan Binhaiwan Central Hospital,Dongguan 523900,China;Guangdong Provincial Center of Human Microecology Engineering and Technology,Guangzhou 524000,China;Guangdong Longsee Biomedical Corporation,Guangzhou 524000,China;Foshan Fourth People’s Hospital,Foshan 528000,China)

机构地区：[1]东莞市滨海湾中心医院,广东东莞523900 [2]广东省人体微生态工程技术研究中心,广东广州524000 [3]广东南芯医疗科技有限公司,广东广州524000 [4]佛山市第四人民医院,广东佛山528000

出　　处：《广东医科大学学报》2021年第4期395-399,共5页Journal of Guangdong Medical University

基　　金：国家自然科学基金青年项目(No.82003786);广州市科技计划项目(No.202002030200);佛山市自筹经费类科技计划项目(医学类科技攻关)(No.1920001000648)。

摘　　要：目的建立同时测定人血清8种叶酸循环代谢物的高效液相色谱串联质谱(LC-MS/MS)法。方法人血清样本沉淀蛋白后挥发干,有机试剂复溶,采用Waters ACQUITY UPLC HSS T3色谱柱、有机试剂梯度洗脱法进行分离,电喷雾正负离子模式下以多反应检测方式检测,内标法定量。结果8种叶酸循环代谢物标准曲线的线性相关系数为0.994~0.999,精密度RSD值为3.9%~12.3%,回收率RSD值为85.4%~113.1%,最低检出限为0.128μg/L。结论LC-MS/MS法具有重现性好、灵敏度高、通量高的特点,可同时测定多种人体叶酸循环代谢物。Objective To establish a LC-MS/MS method for simultaneous detection of 8 folate metabolites in human serum.Methods The protein precipitation extracted from human serum was evaporated and redissolved in organic reagents.The analytes were separated using waters ACQUITY UPLC HSS T3 column and then eluted by gradient elution with organic reagents.The folate metabolites were detected using positive and negative ion electrospray in multiple reaction monitoring(MRM)mode,and the quantification was determined by internal standard method.Results The linear coefficients of 8 folate metabolites were 0.994-0.999.Relative standard deviations of precision and recovery were 3.9%-12.3%and 85.4%-113.1%respectively,and minimum detection limit was 0.128μg/L.Conclusion LC-MS/MS is reproducible,sensitive,and high-throughput for simultaneous detection of multiple folate metabolites in human blood.

关 键 词：高效液相色谱串联质谱 叶酸 循环代谢物 

分 类 号：Q56[生物学—生物化学]