色谱

色谱 ›› 0, Vol. ›› Issue (): 0-0.DOI: 10.3724/SP.J.1123.2012.04010

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直接竞争酶联免疫吸附分析法、高效液相色谱-紫外检测法和高效液相色谱-串联质谱法测定污水中残留克伦特罗

蔡一超,刘曙照   

  1. 扬州大学
  • 收稿日期:2012-04-09 修回日期:2012-05-15 出版日期:2012-05-16 发布日期:2012-05-16
  • 通讯作者: 刘曙照

Determination of clenbuterol residue in waste water by direct competitive enzyme-linked immunosorbent assay,high performance liquid chromatography-UV andhigh performance liquid chromatography-tandem mass spectrometry

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  • Received:2012-04-09 Revised:2012-05-15 Online:2012-05-16 Published:2012-05-16

摘要： 禽畜排泄物中克伦特罗可造成环境水体的污染，本实验分别应用包被抗体直接竞争酶联免疫吸附分析（ELISA）法，高效液相色谱-紫外检测 (HPLC-UV) 法和高效液相色谱串联质谱（HPLC-MS/MS）法测定了污水中克伦特罗的残留量。在优化条件下，ELISA法对克伦特罗标样检测的线性范围为0.001～10 mg/L，线性中浓度为33.20 μg/L，5次重复测定的相对标准偏差(RSD)为8.54%，最低检出浓度为0.763 μg/L。在水样中分别添加克伦特罗浓度水平为0.2、0.02﹑0.002 mg/L，ELISA法测定的平均回收率分别为101.4%、97.6%和105.3%，RSD(n=5)分别为7.9%、6.7%和8.7%。同样前处理条件下，ELISA法、HPLC-UV法和HPLC-MS/MS对污水中残留克伦特罗的最小检出量分别为3.6×10-11 g、2.0×10-9 g和1.0×10-12 g。ELISA法和HPLC-MS/MS法测定屠宰场污水处理池水中克伦特罗残留量分别为0.970 μg/L(n=5，RSD=3.1% )和0.827 μg/L（n=4，RSD=6.2%）。三种测定方法测定结果比较：ELISA法样品前处理和检测方法简单、快速、成本低，可多个样品同时检测，检测灵敏度和精密度符合克伦特罗残留测定要求，特别适合大量样品的快速筛查检测。

关键词: 高效液相色谱, 高效液相色谱-质谱/质谱, 克伦特罗, 污水, 直接竞争酶联免疫吸附分析

Abstract: Clenbuterol in the farm animal would cause pollution to the environmental water. The methods of antibody-immobilized direct competitive Enzyme-Linked Immunosorbent Assay (ELISA), High Performance Liquid Chromatography-UV detector (HPLC-UV) and High Performance Liquid Chromatography-Tandem Mass Spectrometry (HPLC-MS/MS) were employed to determine residual clenbuterol in waste water. Under the optimized conditions, the liner concentrations of ELISA ranged from 0.001 mg/L to 10 mg/L, the middle linerary concentration was 33.20 μg/L with relative standard deviation (RSD) of 8.54%, the minimum detectable concentration was 0.763 μg/L. The clenbuterol average recoveries of ELISA were 101.4%, 97.6% and 105.3% with the RSDs of 7.9%, 6.7% and 8.7% at the spiked level of 0.2, 0.02 and 0.002 mg/L in waste water, the limit of detection (LOD) of ELISA for clenbuterol was 3.6×10-11 g. With the similar sample preparation, the LOD of HPLC-UV and HPLC-MS/MS were 2.0×10-9 g and 1.0×10-12 g. In waste water sample, the residual rlenbuterol was 0.970 μg/L determined by ELISA with RSD of 3.1%, and 0.827 μg/L determined by HPLC-MS/MS with RSD of 6.2%, the result of ELISA was well correspond to that of HPLC-MS/MS. Compare with HPLC-UV and HPLC-MS/MS, ELISA is simple, fast and low cost, it is suitabal for the detection of residual clenbuterol, and especially suitable for the large number of samples screening.

Key words: clenbuterol, direct competitive Enzyme-Linked Immunosorbent Assay (ELISA), High Performance Liquid Chromatography-Tandem Mass Spectrometry (HPLC-MS/MS), High Performance Liquid Chromatography-UV detector (HPLC-UV), Waste water