液相色谱-串联质谱检测血浆3-甲氧酪胺方法的建立

doi:10.3969/j.issn.1673-8640.2017.011.024

检验医学 ›› 2017, Vol. 32 ›› Issue (11): 1046-1050.DOI: 10.3969/j.issn.1673-8640.2017.011.024

• 技术研究与评价_论著 • 上一篇下一篇

液相色谱-串联质谱检测血浆3-甲氧酪胺方法的建立

秦嘉倩, 彭颖斐, 陈方俊, 邵文琦, 吴炯, 张春燕, 王蓓丽, 郭玮, 潘柏申

复旦大学附属中山医院检验科,上海 200032

收稿日期:2017-02-13 出版日期:2017-11-30 发布日期:2017-12-07
作者简介:null
作者简介：秦嘉倩,女,1991年生,技师,主要从事临床生化检验工作。
基金资助:
国家自然基金科学面上项目（81572064）;上海市卫生计生系统重要薄弱学科建设项目（2015ZB0201）

Establishment of a liquid chromatography-tandem mass spectrometry for the determination of plasma 3-methoxytyramine

QIN Jiaqian, PENG Yingfei, CHEN Fangjun, SHAO Wenqi, WU Jiong, ZHANG Chunyan, WANG Beili, GUO Wei, PAN Baishen

Department of Clinical Laboratory,Zhongshan Hospital,Fudan University,Shanghai 200032,China

Received:2017-02-13 Online:2017-11-30 Published:2017-12-07

摘要/Abstract

摘要：

目的建立一种稳定的检测血浆3-甲氧酪胺（3-MT）的液相色谱-串联质谱（LC-MS/MS）方法。方法对LC-MS/MS的分离条件（色谱柱、柱温、pH值）进行优化,建立检测血浆3-MT的方法,并对该方法的线性范围、回收率、精密度、最低检测下限和稳定性进行评价。结果以同位素氘代作为内标,采用BEH HILIC色谱柱进行分离。流动相为100 mmol/L甲酸铵缓冲液（pH值为3）和纯乙腈,梯度洗脱;柱温为35 ℃。LC-MS/ MS检测3-MT的线性范围为5~1 000 pg/mL,定量检出限为5 pg/mL,天间和批间变异系数（CV）分别为<6%和<7%,回收率为97.1%~109.3%。由于3-MT在室温中稳定性较差,因此需冰浴送检。结论建立了检测3-MT的LC-MS/MS方法,能够灵敏、准确地检测血浆3-MT水平。

关键词: 3-甲氧酪胺, 液相色谱-串联质谱法, 嗜铬细胞瘤

Abstract:

Objective To establish a liquid chromatography-tandem mass spectrometry（LC-MS/MS） for plasma 3-methoxytyramine（3-MT） determination. Methods The separation condition of LC-MS/MS（chromatographic column,column temperature and pH value）was improved. The linear range,recovery rate,precision,lower limit of quantitation and stability of LC-MS/MS were evaluated. Results 3-MT was separated by BEH HILIC chromatographic column using 3-MT-d4 as internal standard. The mobile phase consisted of 100 mmol/L ammonium formate aqueous solution（pH value was 3）and acetonitrile. The column temperature was 35 ℃. The linear range of 3-MT determination were 5-1 000 pg/mL. The limit of quantitation was 5 pg/mL. The inter-day and between-run coefficients of variation（CV） were <6% and <7%. The recovery rate was 97.1%-109.3%. As 3-MT was unstable in room temperature,it should be transported under 4 ℃. Conclusions A reliable LC-MS/MS for the determination of 3-MT has been established,with sensitivity and accuracy.

Key words: 3-Methoxytyramine, Liquid chromotography-tandem mass spectrometry, Pheochromocytoma

中图分类号:

R446.1

秦嘉倩, 彭颖斐, 陈方俊, 邵文琦, 吴炯, 张春燕, 王蓓丽, 郭玮, 潘柏申. 液相色谱-串联质谱检测血浆3-甲氧酪胺方法的建立[J]. 检验医学, 2017, 32(11): 1046-1050.

QIN Jiaqian, PENG Yingfei, CHEN Fangjun, SHAO Wenqi, WU Jiong, ZHANG Chunyan, WANG Beili, GUO Wei, PAN Baishen. Establishment of a liquid chromatography-tandem mass spectrometry for the determination of plasma 3-methoxytyramine[J]. Laboratory Medicine, 2017, 32(11): 1046-1050.

图/表 8

参考文献 9

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[5]	EISENHOFER G,LENDERS J W,SIEGERT G,et al.Plasma methoxytyramine:a novel biomarker of metastatic pheochromocytoma and paraganglioma in relation to established risk factors of tumour size,location and SDHB mutation status[J]. Eur J Cancer,2012,48(11):1739-1749.
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[7]	PEITZSCH M,PREJBISZ A,KROIB M,et al.Analysis of plasma 3-methoxytyramine,normetanephrine and metanephrine by ultraperformance liquid chromatography-tandem mass spectrometry:utility for diagnosis of dopamine-producing metastatic phaeochromocytoma[J]. Ann Clin Biochem,2013,50(Pt 2):147-155.
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检测阶段	时间	流速（mL/min）	流动相A（%）	流动相B（%）
1		0.45	5.00	95.00
2	1.00	0.45	30.00	70.00
3	2.00	0.45	30.00	70.00
4	3.00	0.45	40.00	60.00
5	5.00	0.45	5.00	95.00

检测阶段	时间	流速（mL/min）	流动相A（%）	流动相B（%）
1		0.45	5.00	95.00
2	1.00	0.45	30.00	70.00
3	2.00	0.45	30.00	70.00
4	3.00	0.45	40.00	60.00
5	5.00	0.45	5.00	95.00

分析物	转换离子对（m/z）	锥孔电压（kV）	碰撞能量（eV）
3-MT定性	151→119	30	15
3-MT定量	151→91	30	15
3-MT-d4定量	155→95	30	15

分析物	转换离子对（m/z）	锥孔电压（kV）	碰撞能量（eV）
3-MT定性	151→119	30	15
3-MT定量	151→91	30	15
3-MT-d4定量	155→95	30	15

3-MT （ng/mL）	反复冻融（%）	室温放置24 h （%）	自动进样器放置36 h （%）
11.9	6.8	23.7	0.5
18.6	5.5	29.3	0.2

液相色谱-串联质谱检测血浆3-甲氧酪胺方法的建立

Establishment of a liquid chromatography-tandem mass spectrometry for the determination of plasma 3-methoxytyramine

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