基于液相色谱串联质谱技术对血清同型半胱氨酸常规检测系统的正确度评价

doi:10.3969/j.issn.1673-8640.2018.08.003

检验医学 ›› 2018, Vol. 33 ›› Issue (8): 679-682.DOI: 10.3969/j.issn.1673-8640.2018.08.003

基于液相色谱串联质谱技术对血清同型半胱氨酸常规检测系统的正确度评价

钟巧玲¹, 孙骥², 翁旭琦³, 冯萍³, 谢丹芸³

1. 重庆市巴南区第二人民医院检验科,重庆 400054
2.宁波出入境检验检疫局技术中心,浙江宁波 315000
3. 美康生物参考实验室,浙江宁波 315104

收稿日期:2018-03-20 出版日期:2018-08-10 发布日期:2018-09-07
作者简介:null
作者简介：钟巧玲,女,1976年生,学士,副主任技师,主要从事临床免疫检验工作。

Accuracy evaluation of routine measurement procedure for serum homocysteine based on liquid chromatography-tandem mass spectrometry

ZHONG Qiaoling¹, SUN Ji², WENG Xuqi³, FENG Ping³, XIE Danyun³

1. Department of Clinical Laboratory,the Second People's Hospital of Banan District,Chongqing 400054,China
2. Technique Center,Ningbo Entry-Exit Inspection and Quarantine Bureau,Ningbo 315000,Zhejiang,China
3. Reference Laboratory,Medical System Biotechnology Co.,Ltd.,Ningbo 315104,Zhejiang,China

Received:2018-03-20 Online:2018-08-10 Published:2018-09-07

摘要/Abstract

摘要：

目的以同位素稀释液相色谱串联质谱（ID-LC/MS/MS）为比对方法,评价酶循环法测定血清同型半胱氨酸（Hcy）的正确度。方法采用ID-LC/MS/MS和酶循环法同时检测20份新鲜单人份血清中的Hcy浓度,对测定结果进行方法学比对和评价。采用ID-LC/MS/MS和酶循环法同时测定5个水平的工作校准品,根据比对结果调整酶循环法校准品值,直至酶循环法的测定结果与比对方法的测定结果一致。结果采用ID-LC/MS/MS检测SRM 1950标准物质,测定结果与证书标示值一致,相对偏移<1.0%。酶循环法与ID-LC/MS/MS的线性拟合方程为Y=1.204 1X+0.364 6（r²=0.987 9）,平均偏移为24.0%。调整后的酶循环法与ID-LC/MS/MS的线性拟合方程为Y=0.988 6X+0.260 3（r²=0.999 7）,平均偏移为-0.09%。结论 ID-LC/MS/MS能准确测定血清Hcy水平,或许可作为Hcy常规检测系统正确度评价的比对方法。

关键词: 同型半胱氨酸, 同位素稀释液相色谱串联质谱, 酶循环法, 正确度评价

Abstract:

Objective To evaluate the accuracy of enzymatic cycling method for serum homocysteine（Hcy） based on isotope-dilution liquid chromatography-tandem mass spectrometry（ID-LC/MS/MS） as a contrast method.Methods A total of 20 fresh serum samples were determined by enzymatic cycling method and ID-LC/MS/MS,and the results were compared. The levels of serum Hcy were adjusted until the results were consistent,according to the results of 5 working calibrations,which were determined by enzymatic cycling method and ID-LC/MS/MS.Results The results of standard reference material（SRM） 1950 determined by ID-LC/MS/MS were consistent with certificate value,and the relative bias was <1.0%. The linear equation of ID-LC/MS/MS and enzymatic cycling method was Y=1.204 1X+0.364 6（r²=0.987 9）,and the average bias was 24.0%. After adjusting,the linear equation was Y=0.988 6X+0.260 3（r²=0.999 7）,and the average bias was -0.09%.Conclusions The levels of serum Hcy determined by ID-LC/MS/MS are accurate,which can be used for the accuracy evaluation of routine measurement procedure for serum Hcy.

Key words: Homocysteine, Isotope-dilution liquid chromatography-tandem mass spectrometry, Enzymatic cycling method, Accuracy evaluation

中图分类号:

R446.1

钟巧玲, 孙骥, 翁旭琦, 冯萍, 谢丹芸. 基于液相色谱串联质谱技术对血清同型半胱氨酸常规检测系统的正确度评价[J]. 检验医学, 2018, 33(8): 679-682.

ZHONG Qiaoling, SUN Ji, WENG Xuqi, FENG Ping, XIE Danyun. Accuracy evaluation of routine measurement procedure for serum homocysteine based on liquid chromatography-tandem mass spectrometry[J]. Laboratory Medicine, 2018, 33(8): 679-682.

图/表 6

表1 ID-LC/MS/MS检测NIST SRM 1950标准物质的结果

批次	实测值（μmol/L）	变异系数（%）	相对偏移（%）
1	8.55	1.65	0.59
2	8.49	1.35	0.12

图1 酶循环法与ID-LC/MS/MS测定结果的比较

图2 酶循环法与ID-LC/MS/MS的Bland-Altman偏移图

表2 酶循环法调整后与ID-LC/MS/MS测定结果的比较

工作校准品编号	酶循环法（μmol/L）	ID-LC/MS/MS（μmol/L）	偏移（%）
Ⅰ	9.05	9.19	-1.58
Ⅱ	17.94	17.78	0.87
Ⅲ	23.02	22.72	1.30
Ⅳ	34.03	34.06	-0.10
Ⅴ	48.42	48.89	-0.96

图3 酶循环法调整后与ID-LC/MS/MS法结果比较

图4 调整后的酶循环法与ID-LC/MS/MS的Bland-Altman偏移图

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基于液相色谱串联质谱技术对血清同型半胱氨酸常规检测系统的正确度评价

Accuracy evaluation of routine measurement procedure for serum homocysteine based on liquid chromatography-tandem mass spectrometry

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