食品科学 ›› 2010, Vol. 31 ›› Issue (12): 169-173.doi: 10.7506/spkx1002-6630-201012039

• 分析检测 • 上一篇    下一篇

特布他林残留的酶联免疫检测方法的建立

孙海新1,凌红丽1 ,* ,张玉兰2,郗日沫2,李志中1,高亚东1,董瑞娥1,赵玉惠1   

  1. 1.青岛康地恩药业有限公司
    2.山东大学化学与化工学院
  • 收稿日期:2009-09-21 出版日期:2010-06-15 发布日期:2010-12-29
  • 通讯作者: 凌红丽 E-mail:hollymxx@163.com
  • 基金资助:

    青岛市科技计划项目- 食品安全专项(06-3-2-4-nsh);国家“863”计划项目(07AA10Z435)

Development of an Indirect Competitive ELISA for Determination of Terbutaline Residue in Pork

SUN Hai-xin1,LING Hong-li1,*,ZHANG Yu-lan2,XI Ri-mo2,LI Zhi-zhong1,GAO Ya-dong1,DONG Rui-e1,ZHAO Yu-hui1   

  1. 1. Qingdao Continent Pharmaceutical Group, Qingdao 266111, China;
    2. School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100, China
  • Received:2009-09-21 Online:2010-06-15 Published:2010-12-29
  • Contact: LING Hong-li1,*, E-mail:hollymxx@163.com

摘要:

采用自主制备的特布他林特异性抗体建立特布他林残留的间接竞争酶联免疫(ELISA)检测方法,对该检测体系的灵敏度、准确度、精密度、特异性进行测定,并将该检测方法与高效液相色谱(HPLC)法进行对比分析。结果表明:特布他林残留检测体系的检测范围为1~100ng/mL,灵敏度为0.47ng/mL,检测限为1ng/mL,回收率80%~99%,与盐酸克伦特罗、硫酸沙丁胺醇的交叉反应率分别为94.9%、93.9%,与盐酸莱克多巴胺及肾上腺素的交叉反应率小于0.01%。采用HPLC 法进行沙丁胺醇残留的检测时,其检测范围为10~200μg/mL,检测限为1μg/mL,回收率为79.2%~94.8%。ELISA 法与HPLC 法相比,灵敏度较高、特异性强、检测结果准确度相近,但在检测结果稳定性方面逊于HPLC 法。

关键词: 特布他林, 药物残留, 酶联免疫, 高效液相色谱

Abstract:

The specific antibodies against terbutaline prepared in our previous studies were used to develop an indirect competitive ELISA for the determination of terbutaline residue in pork. The sensitivity, accuracy, precision and specificity of the method were measured, followed by a comparison with HPLC method. The developed ELISA exhibited good linearity over the concentration range from 1 to 100 ng/mL, a sensitivity of 0.47 ng/mL, a limit of detection of 1 ng/mL and a spike recovery range from 80% to 99%. Cross-reactivities between the antibodies against terbutaline prepared and clenbutarol or salbutamol were 94.9% and 93.9%, respectively, and those between the antibodies against terbutaline prepared and ractopamine or adrenaline were both less than 0.01%. The developed HPLC method exhibited a linear range from 10 to 200μg/mL, a limit of detection of 1μg/mL and a spike recovery range from 79.2% to 94.8%. The developed ELISA exhibited higher sensitivity and specificity than the developed HPLC, whereas the stability of the former was inferior to that of the latter. Their accuracies were similar.

Key words: terbutaline, residue, ELISA, HPLC

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