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反相液相色谱-串联质谱法测定尿儿茶酚胺和甲氧基肾上腺素

[Determination of urine catecholamines and metanephrines by reversed-phase liquid chromatography-tandem mass spectrometry].

作者信息

Cheuk Man-Yung, Lo Yun-Chuen, Poon Wing-Tat

机构信息

Department of Pathology, Pamela Youde Nethersole Eastern Hospital, Hong Kong 999077, China.

出版信息

Se Pu. 2017 Oct 8;35(10):1042-1047. doi: 10.3724/SP.J.1123.2017.06011.

DOI:10.3724/SP.J.1123.2017.06011
PMID:29048800
Abstract

The measurement of urine catecholamine and metanephrine concentrations is important for biochemical screening and diagnosis of pheochromocytoma. The goal of this work was to develop a simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determining catecholamines and metanephrines in urine to replace an existing liquid chromatographic method using electrochemical detection. Urine samples were prepared using Oasis weak-cation-exchange cartridges. The eluate was analyzed on an Agilent ZORBAX Eclipse Plus Phenyl-Hexyl column in 3 min. Adrenaline, noradrenaline, dopamine, metanephrine, normetanephrine, and their deuterated internal standards were monitored in positive electrospray ionization mode by multiple reaction monitoring (MRM). No evidence of ion suppression was observed. The assay was linear up to 5μmol/L for adrenaline, 5μmol/L for noradrenaline, 6.1μmol/L for dopamine, 5.6μmol/L for metanephrine, and 34.6μmol/L for normetanephrine, with lower limits of quantification of 5, 5, 12, 6 and 7 nmol/L, respectively. The intra-day and inter-day precisions for all analytes ranged from 0.59% to 4.64% and 1.98% to 4.80%, respectively. External quality assurance samples were assayed and showed excellent agreement with the target values. This simple method provides an improved assay for determining urine catecholamines and metanephrines.

摘要

尿儿茶酚胺和甲氧基肾上腺素浓度的测定对于嗜铬细胞瘤的生化筛查和诊断至关重要。本研究的目的是开发一种简单的液相色谱-串联质谱(LC-MS/MS)方法来测定尿中的儿茶酚胺和甲氧基肾上腺素,以取代现有的电化学检测液相色谱法。使用Oasis弱阳离子交换柱制备尿样。洗脱液在Agilent ZORBAX Eclipse Plus苯基己基柱上3分钟内进行分析。通过多反应监测(MRM)在正电喷雾电离模式下监测肾上腺素、去甲肾上腺素、多巴胺、甲氧基肾上腺素、去甲氧基肾上腺素及其氘代内标。未观察到离子抑制现象。该测定法对肾上腺素的线性范围高达5μmol/L,对去甲肾上腺素为5μmol/L,对多巴胺为6.1μmol/L,对甲氧基肾上腺素为5.6μmol/L,对去甲氧基肾上腺素为34.6μmol/L,定量下限分别为5、5、12、6和7 nmol/L。所有分析物的日内和日间精密度分别为0.59%至4.64%和1.98%至4.80%。对外部质量保证样品进行了分析,结果与目标值高度一致。这种简单的方法为测定尿儿茶酚胺和甲氧基肾上腺素提供了一种改进的检测方法。

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