液相色谱-串联质谱法检测血清25-羟基维生素D的方法建立和性能评价  

Establishment on a LC-MS/MS and its performance characteristic evaluation for the determination of serum 25-hydroxyvitamin D

作　　者：宋斌斌[1] 秦嘉倩 彭颖斐[1] 张春燕 吴炯[1] 王蓓丽[1] 郭玮[1] 潘柏申[1] 

机构地区：[1]复旦大学附属中山医院检验科,上海200032

出　　处：《检验医学》2015年第5期416-421,共6页Laboratory Medicine

基　　金："十二五"国家科技支撑计划课题资助项目(2012BAI37B01);国家临床重点检验专科建设项目资助项目

摘　　要：目的：建立定量检测血清25－羟基维生素 D[25（OH）D]的液相色谱－串联质谱（LC－MS／MS）方法，并对此方法进行基本分析性能验证。方法使用 Waters？ XevoTM TQ MS ACQUITY UPLC？液质联用仪（LC－MS／MS）同时定量检测血清中的25（OH）D2和25（OH）D3含量。参照美国食品和药品管理局（FDA）的生物分析方法验证导则标准对所建方法进行线性、检出限、精密度和准确性等基本性能的验证。结果LC－MS／MS检测25（OH）D2和25（OH）D3的线性范围为6．25～500．00 nmol／L；25（OH）D2和25（OH）D3的定量检出限分别为2．50和1．25 nmol／L；批内、批间变异系数（CV）分别为＜4％、＜6％；回收率为93．26％～112．16％；维生素D 国际室间质量评价计划（DEQAS）室间质评结果偏倚＜10％。结论建立的 LC－MS／MS 基本性能符合评价标准，能够灵敏且准确的检测出血清中25（OH）D2和25（OH）D3的浓度。Objective To establish a liquid chromatography-tandem mass spectrometry （ LC-MS/MS） for the quantitation determination of serum 25-hydroxyvitamin D [25（OH） D] and perform the general method performance verification.Methods The LC-MS/MS was used for the quantitation determination of 25（OH）D2 and 25（OH）D3 in serum by Waters？ XevoTM TQ MS ACQUITY UPLC？ mass spectrometry instrument.According to US Food and Drug Administration （ FDA）′s guideline （ Guidance for Industry Bioanalytical Method Validation）, the general method performance verification was performed for linearity, determination limit, precision and accuracy.Results The linear ranges of 25（OH）D2 and 25（OH）D3 were 6.25-500.00 nmol/L.The LC-MS/MS had quantitation determination limits of 2.50 nmol/L for 25（OH） D2 and 1.25 nmol/L for 25（OH） D3.The within-run and between-run coefficients of variation （CV） of 25（OH）D2 and 25（OH）D3 were 〈4% and 〈6%, respectively.The recovery rates were 93.26%-112.16%.The result of the Vitamin D External Quality Assessment Scheme （ DEQAS） interlaboratory quality assessment had a bias 〈10%.Conclusions The basic performance of LC-MS/MS meets the evaluation standards, and LC-MS/MS is sensitive and accurate for detecting the concentrations of 25（OH）D2 and 25（OH）D3 in serum.

关 键 词：25羟基维生素D 液相串联质谱 性能验证 

分 类 号：Q565[生物学—生物化学]