色谱

色谱 ›› 2010, Vol. 28 ›› Issue (08): 759-764.DOI: 10.3724/SP.J.1123.2010.00759

• 研究论文 • 上一篇    下一篇

高效液相色谱-串联质谱法检测动物尿液中的15种β-受体激动剂

聂建荣*, 朱铭立, 连槿, 潘云山, 邓香连, 胡翠萍   

  1. 广州市农业标准与监测中心, 广东 广州 510315
  • 收稿日期:2010-04-16 修回日期:2010-06-21 出版日期:2010-08-28 发布日期:2010-08-28
  • 通讯作者: 聂建荣,硕士,高级工程师,研究方向为兽药残留分析.

Determination of fifteen β-agonists in animal urine by high performance liquid chromatography-tandem mass spectrometry

NIE Jianrong*, ZHU Mingli, LIAN Jin, PAN Yunshan, DENG Xianglian, HU Cuiping   

  1. Guangzhou Agriculture Standard and Supervisory Center, Guangzhou 510315, China
  • Received:2010-04-16 Revised:2010-06-21 Online:2010-08-28 Published:2010-08-28

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摘要： 建立了高效液相色谱-串联质谱检测动物尿液中克仑特罗、莱克多巴胺、沙丁胺醇、西马特罗、马布特罗、妥布特罗、班布特罗、马贲特罗、塞布特罗、齐帕特罗、福莫特罗、氯丙那林、特布他林、喷布特罗和溴布特罗等15种β-受体激动剂的分析方法。样品用高氯酸溶液酸解及沉淀蛋白质,经HLB固相萃取小柱净化、富集后,以甲醇和0.1%(体积分数,下同)甲酸水溶液作为流动相进行梯度洗脱,采用多反应监测(MRM)模式进行定性和定量分析。15种β-受体激动剂在0.25～20 μg/L的范围内线性关系良好(r≥0.9995)。在0.25、1.0、10 μg/L添加水平下的回收率为62.1%～107%,相对标准偏差(n=10)为3.5%～9.9%,定量限(以信噪比＞10计)为0.25 μg/L。该方法精密度好,灵敏度高,能简便、快速、准确地测定动物尿液中的15种β-受体激动剂。

关键词: β-受体激动剂, 动物尿液, 高效液相色谱-质谱法

Abstract: A high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was established for the determination of fifteen β-agonists (clenbuterol, ractopamine, salbutamol, cimaterol, mabuterol, tulobuterol, bambuterol, mapenterol, cimbuterol, zilpaterol, formoterol, clorprenaline, terbutaline, penbutolol and brombuterol) in animal urine. Perchloric acid solution was used to acidify the sample and precipitate protein in the sample. The sample was purified and concentrated by an HLB mini-column. The separation of the β-agonist was performed on an Agilent 1100 HPLC system with a Eclipse XDB-C18 column by using gradient elution with methanol and water (containing 0.1% (v/v) formic acid) as the mobile phases at a flow rate of 1 mL/min. Qualitative and quantitative analysis of the fifteen β-agonists, which were ionized by electrospray ionization interface (ESI), were carried out in multiple reaction monitoring (MRM) mode with API 4000 tandem mass spectrometry. The calibration curves showed good linearity in the mass concentration range of 0.25-20 μg/L with the correlation coefficients r≥0.9995. The recoveries of the fifteen β-agonists ranged from 62.1% to 107% at the spiked levels of 0.25, 1.0 and 10 μg/L. The relative standard deviations (n=10) were between 3.5% and 9.9%. The limits of quantification (S/N＞10) were 0.25 μg/L for all the analytes. This method is simple, rapid, sensitive and accurate.

Key words: β-agonists, animal urine, high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS)