[1]李 敏,于洪远,李会强.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[J].现代检验医学杂志,2024,39(01):136-140.[doi:10.3969/j.issn.1671-7414.2024.01.025]
　LI Min,YU Hongyuan,LI Huiqiang.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[J].Journal of Modern Laboratory Medicine,2024,39(01):136-140.[doi:10.3969/j.issn.1671-7414.2024.01.025]
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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()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第39卷
期数:
2024年01期
页码:
136-140
栏目:
研究简报·实验技术
出版日期:
2024-01-15

文章信息/Info

Title:
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
文章编号:
1671-7414（2024）01-136-05
作者:
李 敏1; 于洪远1; 李会强2
（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）
Author(s):
LI Min1;  YU Hongyuan1;  LI Huiqiang2
（1. Beijing Aerospace General Hospital, Beijing 100076, China; 2. School of Laboratory Medicine, Tianjin Medical University, Tianjin 300203, China）
关键词:
25 羟维生素D3; 候选参考测量程序; 性能评价
分类号:
R446.112；Q503
DOI:
10.3969/j.issn.1671-7414.2024.01.025
文献标志码:
A
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 andLaboratory 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 interbatch 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.

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备注/Memo

备注/Memo:
作者简介： 李敏（1991-），女，本科，主管技师，研究方向：临床生化检验标准化，E-mail：1140368364@qq.com。
通讯作者：李会强，男，博士生导师，E-mail：lihuiqiang1965@163.com。
更新日期/Last Update: 2024-01-15