液相色谱串联质谱法检测微量血维生素D含量  

The Measurement of Vitamin D in Trace Amount of Blood by Liquid Chromatography Tandem-Mass Spectrometry

作　　者：战思恩[1] 李幽然 刘颖 贾妍 翟燕红[1] 曹正 ZHAN Si-en;LI You-ran;LIU Ying;JIA Yan;ZHAI Yan-hong;CAO Zheng(Department of Clinical Laboratory,Beijing Obstetrics and Gynecology Hospital,Capital Medical University,Beijing 100026,China)

机构地区：[1]首都医科大学附属北京妇产医院检验科

出　　处：《标记免疫分析与临床》2019年第8期1366-1369,1427,共5页Labeled Immunoassays and Clinical Medicine

基　　金：首都医科大学附属北京妇产医院中青年学科骨干培养专项(编号:FCYY201708)

摘　　要：目的建立液相色谱串联质谱法(LC- MS/MS法)测定微量血维生素D含量的方法。方法采用非衍生处理方法,利用正离子电喷雾离子化(ESI+)、多反应监测模式(MRM)、同位素内标法检测微量血维生素D含量并进行相关方法学验证。结果 vitD2在5~100 ng/mL、vitD3在5~150 ng/mL范围内线性良好,线性相关系数分别为0.9988、0.9984。vitD2和vitD3的批内精密度和批间精密度变异系数均小于10%。vitD2低浓度回收率为87.58%,高浓度为94.68%;vitD3的低浓度回收率为92.62%,高浓度为87.65%。vitD2、vitD3的LOD分别为0.36 ng/mL和1.33 ng/mL,LOQ分别为2.5 ng/mL和2.5 ng/mL。患者样本在4℃的稳定性较好,可至少保存7d。vitD2和vitD3的携带污染分别为0.06%和 0.07%,满足要求。结论此LC- MS/MS检测法不仅样本用量少,而且操作简单,无污染,更适合临床检测分析。Objective To establish a method for the measurement of vitamin D in trace amount of blood by liquid chromatography- tandem mass spectrometry(LC- MS/MS). Methods A non- derivative method was used to detect the content of vitamin D by LC- MS / MS through the positive electrospray ionization(ESI+)and multiple reaction monitor(MRM)mode. Results The linearity was good for vitD2 within range of 5-100ng/mL and for vitD3 within 5-150ng/mL,and the linear correlation coefficients were 0.9988 and 0.9984 ,respectively.All the intra- and inter- batch precision variation coefficients for both vitD2 and vitD3 were less than 10 %.The low and high concentration recovery rates of vitamin vitD2 were 87.58 % and 94.68%,respectively;The low and high concentration recovery rate of vitamin vitD3 were 92.62 % and 87.65% respectively.The LODs of vitD2 and vitD3 were 0.36 ng/mL and 1.33 ng/mL,respectively,and the LOQ of vitD2 and vitD3 were 2.5 ng/mL and 2.5 ng/mL,respectively.The patient samples were stable at 4℃ and could be preserved for at least 7 days.The carryover of vitD2 and vitD3 were 0.06% and 0.07%, respectively. Conclusion The LC- MS/MS method in this study only needs micro- volume sample,and it is simple to operate and pollution- free.So it is suitable for the clinical analysis.

关 键 词：维生素D2 维生素D3 液相色谱串联质谱 微量血 

分 类 号：R446.1[医药卫生—诊断学]