基于同位素稀释液相色谱串联质谱技术检测血清25(OH)D3的候选参考测量程序的建立及性能评价
Establishment and Performance Evaluation of Candidate Reference Measurement Procedures for the Detection of Serum 25(OH)D3 Based on Isotope Dilution Liquid Chromatography Tandem Mass Spectrometry
李敏 1于洪远 1李会强2
作者信息
- 1. 北京航天总医院检验科,北京 100076
- 2. 天津医科大学医学检验学院,天津 300203
- 折叠
摘要
目的 建立基于同位素稀释液相色谱串联质谱技术(liquid phase chromatography tandem mass spectrometry,LC-MS/MS)的血清 25 羟维生素D3[25-hydroxy vitamin D3,25(OH)D3]的候选参考测量程序.方法 采用同位素标准溶液为内标,液液萃取进行前处理,正离子电喷雾模式进行监测.参照美国临床实验室标准化协会(Clinical and Laboratory Standards Institute,CLSI)C62-A和EP15-A3 等文件,对方法的正确度、精密度、线性范围、定量限、检测下限、相对基质效应等进行验证.采用候选参考测量程序和质谱常规程序检测 40 份临床血清样本,评价两种方法的一致性.结果 候选方法的分析时间为15 min,通过色谱的等度洗脱可有效分离同分异构体3-epi-25(OH)D3,特异度好.测定参考实验室室间比对(RELA)样本,偏倚低于1.5%.批内精密度和批间精密度分别为0.75%～2.31%和1.28%～2.01%.定量限和检测下限分别为0.85 ng/ml和1.84 ng/ml.在2.5～220ng/ml浓度范围内线性良好.无相对基质效应和携带污染.质谱常规程序与候选参考程序的相关性较好(r=0.982),但低浓度样本处的偏差超出卫健委临检中心室间质评的允许总误差±25%.结论 成功建立了基于LC-MS/MS技术的血清 25(OH)D3 的候选参考测量程序,分析性能符合要求,可用于临床常规方法的量值溯源.
Abstract
Objective To establish a candidate reference measurement procedure for the detection of serum 25-hydroxy vitamin D3[25(OH)D3]based on isotope dilution liquid chromatography tandem mass spectrometry(LC-MS/MS).Methods Isotope standard solution was used as internal standard,liquid-liquid extraction was used for pre-treatment,and positive ion electrospray ionization mode was used for monitoring.The accuracy,precision,linear range,limit of quantitation,detection limit and relative matrix effect of method were verified based on documents of the America Clinical and Laboratory Standards Institute(CLSI)such as C62-A and EP15-A3.Candidate reference measurement procedure and mass spectrometry routine procedure were used to detect 40 clinical serum samples,and to evaluate the consistency of the two methods.Results The analysis time of the candidate method was 15 min.Isometric elution of chromatography was used to effectively separate the isomer 3-epi-25(OH)D3,with good specificity.RELA comparison sample was measured,with a bias of less than 1.5%.The intra-batch precision and inter-batch precision ranged from 0.75%to 2.31%and 1.28%to 2.01%,respectively.The limits of quantification and detection were 0.85 ng/ml and 1.84 ng/ml.It had good linearity in the concentration range of 2.5～220 ng/ml,and there was no relative matrix effect and carrier contamination.The correlation between the mass spectrometry routine procedure and candidate reference procedure was good(r=0.982),while the deviation at low concentration samples exceeded the allowable total error±25%in the external quality assessment of the National Center for Clinical Laboratories.Conclusion A candidate reference measurement procedure for serum 25(OH)D3 technology based on LC-MS/MS was successfully established,and the analytical performance met the requirements,which could be used for quantitative traceability by clinical conventional methods.
关键词
25羟维生素D3/候选参考测量程序/性能评价Key words
25-hydroxyvitamin D3/candidate reference measurement procedures/performance evaluation引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
维普
万方数据