色谱

色谱

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高效液相色谱-离子阱质谱法测定尿液中β2-受体激动剂及β-受体阻断剂

苗虹1, 邹建宏3, 范赛1,2, 甘乐文3, 赵云峰1, 吴永宁1,2*   

  1. 1. 中国疾病预防控制中心营养与食品安全所, 北京 100021; 2. 中国农业大学动物医学院, 北京 100193; 3. 中国人民解放军第二炮兵总医院, 北京 100088
  • 收稿日期:2010-01-07 修回日期:2010-03-07 出版日期:2010-06-28 发布日期:1980-06-25
  • 通讯作者: 吴永宁

Determination of β2-agonists and β-blockers in urine using high performance liquid chromatography-ion trap mass spectrometry

MIAO Hong1, ZOU Jianhong3, FAN Sai1,2, GAN Lewen3, ZHAO Yunfeng1, WU Yongning1,2*   

  1. 1. Institute of Nutrition and Food Safety, Chinese Centre for Disease Control and Prevention, Beijing 100021, China; 2. College of Veterinary Medicine, China Agricultural University, Beijing 100193, China; 3. General Hospital of the Second Artillery, the Chinese People’s Liberation Army, Beijing 100088, China
  • Received:2010-01-07 Revised:2010-03-07 Online:2010-06-28 Published:1980-06-25
  • Contact: WU Yongning

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摘要： 建立了尿液中23种β2-受体激动剂及5种β-受体阻断剂的高效液相色谱-离子阱质谱(HPLC-IT-MS)测定方法。尿液样品采用冷冻高速离心沉淀蛋白,上清液过ExtrelutTM硅藻土柱,用乙酸乙酯洗脱后,洗脱液经旋转蒸发仪浓缩并复溶待测。HPLC分离采用AtlantisT3-150 mm色谱柱,以甲醇和含0.1%甲酸的水溶液为流动相梯度洗脱,IT-MS采用电喷雾离子源在多反应离子监测模式下测定。定量分析选择9种经过氘代同位素标记的β2-受体激动剂为内标。各化合物的线性范围为0.005～0.16 mg/L,尿液中的检出限均能达到0.2 μg/L。空白尿液样品中不同加标水平的回收率为57.1%～127.1%,相对标准偏差为1.1%～31.1%。该方法简便快速,灵敏度高,适用于人或动物尿液中23种β2-受体激动剂及5种β-受体阻断剂的定性和定量分析。

关键词: β2-受体激动剂, β-受体阻断剂, 高效液相色谱-质谱法, 尿液

Abstract: A method has been developed for the determination of 23 β2-agonists and 5 β-blockers in urine samples using high performance liquid chromatography-ion trap mass spectrometry (HPLC-IT-MS). Urine samples were first deproteinized by high-speed frozen centrifugation, and the supernatants were loaded on an ExtrelutTM diatomite column for clean-up. The analytes were eluted by ethyl acetate and concentrated for further analysis. The analytical separation was performed on an AtlantisT3-150 mm chromatographic column with the gradient elution using methanol and water (containing 0.1% formic acid). The detection was carried on a linear ion trap mass spectrometer under multiple reaction monitoring (MRM) mode with the source operated in positive mode of electrospray ionization (ESI+). Nine deuterium labeled β2-agonists were used as internal standards for quantitative analysis. The results showed that the linear ranges for 23 β2-agonists and 5 β-blockers were 0.005－0.16 mg/L, and the limits of detection were all around 0.2 μg/L. The mixed standard solution was added into the blank urine samples, and the recoveries of 23 β2-agonists and 5 β-blockers were ranged from 57.1% to 127.7% with the relative standard deviations of 1.1%-31.1%. The results demonstrate that the method is easy, fast, sensitive, and suitable for the confirmation and quantification of 23 β2-agonists and 5 β-blockers in urine samples.

Key words: urine , β2-agonists, β-blockers, high performance liquid chromatography-mass spectrometry (HPLC-MS)