检验医学 ›› 2015, Vol. 30 ›› Issue (5): 433-436.DOI: 10.3969/j.issn.1673-8640.2015.05.006

• 质谱技术在临床生化检验中的应用专题 • 上一篇    下一篇

尿儿茶酚胺LC-MS/MS检测方法的建立

彭颖斐, 吴炯, 郭玮, 陈方俊, 秦嘉倩, 徐雯, 潘柏申   

  1. 复旦大学附属中山医院检验科,上海 200032
  • 收稿日期:2015-01-31 出版日期:2015-05-30 发布日期:2015-06-17
  • 作者简介:null

    作者简介:彭颖斐,女,1985年生,学士,技师,主要从事临床检验工作。

    通讯作者:潘柏申,联系电话:021-64041990-2376。

  • 基金资助:
    “十二五”国家科技支撑计划资助项目(2012BAI37B01);国家临床重点检验专科建设资助项目

Establishment of a LC-MS/MS for the determination of urinary catecholamines

PENG Yingfei, WU Jiong, GUO Wei, CHEN Fangjun, QIN Jiaqian, XU Wen, PAN Baishen   

  1. Department of Clinical Laboratory, Zhongshan Hospital, Fudan University, Shanghai 200032, China
  • Received:2015-01-31 Online:2015-05-30 Published:2015-06-17

摘要: 目的 建立尿液儿茶酚胺[包括肾上腺素(E)、去甲肾上腺素(NE)和多巴胺(DA)]的液相色谱-串联质谱(LC-MS/MS)检测方法。方法 以E-d3、NE-d6和DA-d4盐酸盐为内标,采用WATERS ACQUITY UPLC® HSS T3色谱柱(2.1 mm×100 mm,1.8 μm)进行分离。流动相A1为2 ‰甲酸水溶液,流动相B1为甲醇(纯度为99.9%),流速为0.5 mL/min,按A1∶B1=95∶5等度洗脱。采用电喷雾(ESI)正离子模式进行质谱分析。对方法的线性、准确度(回收率)、不精密度及最低检出限等进行性能验证。选取表面健康人群65名,建立E、NE和DA的参考区间。结果 采用建立的LC-MS/MS检测尿液E、NE和DA,保留时间分别为0.58、0.89和0.63 min,检测范围为0.50~800.00 ng/mL。E、NE和DA的加标平均回收率分别为97.95%、97.78%和101.03%,最低检出限分别为0.25、2.50和2.50 ng/mL。E、NE和DA的参考区间分别为5.4~25.8、 28.5~73.1和269.1~420.5 μg/24 h。结论 建立的LC-MS/MS方法能同时检测尿液中的E、NE和DA,可为嗜铬细胞瘤的诊断提供可靠的信息。

关键词: 肾上腺素, 去甲肾上腺素, 多巴胺, 尿液, 液相色谱-串联质谱

Abstract: Objective To establish a liquid chromatography-tandem mass spectrometry(LC-MS/MS) for the determination of urinary catecholamines, including epinephrine(E), norepinephrine(NE) and dopamine(DA). Methods E, NE and DA were separated by WATERS ACQUITY UPLC® HSS T3 (2.1 mm×100 mm,1.8 μm column) using E-d3, NE-d6 and DA-d4 as internal standards. The mobile phase consisted of 2‰ formic acid aqueous solution (A1) and methyl alcohol (B1, purity=99.9%). The flow rate of 0.50 mL/min was applied to the chromatographic column. Flow starting was performed with A1∶B1=95∶5. Electrospray ionization (ESI) was operated in positive ion mode. The LC-MS/MS was used, with linearity, accuracy (recovery rate), imprecision and lower limit of determination being evaluated. A total of 65 healthy subjects were enrolled, and the reference ranges for E, NE and DA were established. Results The retention times of E, NE and DA were 0.58, 0.89, 0.63 min, respectively. The determination range was from 0.50 ng/mL to 800.00 ng/mL. The mean recovery ranges were 97.95%, deter mination range 97.78% and 101.03%. The lower limits of determination for E, NE and DA were 0.25, 2.50 and 2.50 ng/mL. The reference ranges were 5.4-25.8 μg/24 h for E, 28.5-73.1 μg/24 h for NE and 269.1-420.5 μg/24 h for DA. Conclusions The LC-MS/MS for the simultaneous determination of E, NE and DA has been established and is suitable for clinical application. It can provide reliable information for the diagnosis of phaeochromocytoma.

Key words: Epinephrine, Norepinephrine, Dopamine, Urine, Liquid chromatography-tandem mass spectrometry

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