血清醛固酮候选参考测量程序的建立及性能评估

doi:10.3969/j.issn.1673-8640.2020.08.016

摘要/Abstract

摘要：

目的建立基于同位素稀释液相色谱-串联质谱（ID-LC-MS/MS）的血清醛固酮候选参考方法。方法以甲基叔丁基醚为萃取剂,采用液液萃取方式进行样本前处理。以醛固酮-²H₇为内标,采用电喷雾离子源（ESI）负离子模式建立检测血清醛固酮的ID-LC-MS/MS方法,并对方法的准确度、精密度、灵敏度、线性等基本分析性能进行验证。结果 ID-LC-MS/MS测定血清醛固酮的线性范围为0.08~3.55 nmol/L,检测限和定量限（LOQ）分别为0.012 nmol/L和0.045 nmol/L。批内和批间变异系数（CV）分别为<5.0%和<4.1%。平均加标回收率为98.52%~100.14%。测定国际临床化学和检验医学联合会参考实验室外部质量评价计划（RELA）样本的结果偏移<1.6%。结论建立的测定血清醛固酮的ID-LC-MS/MS方法准确、精密,有望作为测定血清醛固酮的参考方法。

关键词: 醛固酮, 同位素稀释液相色谱-串联质谱, 参考方法

Abstract:

Objective To establish a candidate reference measurement procedure for serum aldosterone by isotope dilution liquid chromatography-tandem mass spectrometry（ID-LC-MS/MS）. Methods The sample was prepared by liquid-liquid extraction with methyltert-bytylether as extractant. The aldosterone-²H₇ was selected as internal standard. Samples were prepared by liquid-liquid extraction,and aldosterone was detected by ID-LC-MS/MS in negative ion mode. The basic analysis performance of the established candidate reference measurement procedure,including accuracy,precision,sensitivity and linearity,was evaluated. Results The linear range of aldosterone was 0.08-3.55 nmol/L. The limit of detection and the limit of quantitation were （LOQ）0.012 nmol/L and 0.045 nmol/L,respectively. The within-run coefficients of variation（CV） was <5.0%,and the between-run CV was <4.1%. The average recovery rates were 98.52%-100.14%. The bias of the International Federation of Clinical Chemistry and Laboratory Medicine External Quality Assessment Scheme for Reference Laboratories in Laboratory Medicine（RELA） was <1.6%. Conclusions The established ID-LC-MS/MS for detecting aldosterone in serum is accuracy and precise,and may be used as a candidate reference measurement procedure.

Key words: Aldosterone, Isotope dilution liquid chromatography-tandem mass spectrometry, Reference method

中图分类号:

R446.1

邹继华, 李全乐, 沈敏, 张曼, 杨晓东, 屠敏敏, 冯东方, 孙宏娜. 血清醛固酮候选参考测量程序的建立及性能评估[J]. 检验医学, 2020, 35(8): 811-817.

ZOU Jihua, LI Quanle, SHEN Min, ZHANG Man, YANG Xiaodong, TU Minmin, FENG Dongfang, SUN Hongna. Establishment and performance evaluation of candidate reference measurement procedure for serum aldosterone by liquid chromatography-tandem mass spectrometry[J]. Laboratory Medicine, 2020, 35(8): 811-817.

图/表 12

参考文献 12

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名称	测定方式	母离子/（m/z）	子离子/（m/z）	驻留时间/ms	去簇电压/V	碰撞电压/V
醛固酮	定量	359.2	189.1	270	-110	-25
	定性	359.2	331.2	270	-110	-22
醛固酮-²H₇	定量	366.2	194.1	270	-110	-25
	定性	366.2	338.2	270	-110	-21

名称	测定方式	母离子/（m/z）	子离子/（m/z）	驻留时间/ms	去簇电压/V	碰撞电压/V
醛固酮	定量	359.2	189.1	270	-110	-25
	定性	359.2	331.2	270	-110	-22
醛固酮-²H₇	定量	366.2	194.1	270	-110	-25
	定性	366.2	338.2	270	-110	-21

样本类型	样本数	醛固酮/（nmol/L）	CI/QI比值平均值（范围）
标准溶液	63	0.08~3.55	0.71（0.62~0.77）
血清	90	0.11~3.15	0.69（0.61~0.78）

样本类型	样本数	醛固酮/（nmol/L）	CI/QI比值平均值（范围）
标准溶液	63	0.08~3.55	0.71（0.62~0.77）
血清	90	0.11~3.15	0.69（0.61~0.78）

样本编号	理论值/（nmol/L）	样本数	测定均值/（nmol/L）	平均R_SN	CV/%	偏移/%
1	0.075	10	0.083	39	6.32	5.60
2	0.056	10	0.064	27	8.69	10.28
3	0.045	10	0.052	15	13.88	12.56