Establishment of Enhanced Chemiluminescent Immunoassay Formats for Stanozolol Detection in animal-derived foodstuffs and Other Matrices

Establishment of Enhanced Chemiluminescent Immunoassay Formats for Stanozolol Detection in animal-derived foodstuffs and Other Matrices
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建立用于动物源性食品和其他基质中康力龙检测的增强化学发光免疫分析方法

DOI:
10.1007/s12161-015-0307-3
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发表时间:
2016-05
影响因子:
2.9
通讯作者:
Xi Rimo*
Xi Rimo*
中科院分区:
农林科学3区
文献类型:
--
作者:
Song, ZhaoRui;Yin, Yongmei;Meng, Meng;Xi Rimo*

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司他唑尔是应用最广泛的合成代谢类固醇之一,但也有许多副作用。有必要开发一种快速、灵敏的斯坦诺唑尔筛选方法来监测其滥用情况。本研究制备了抗斯坦诺唑尔的多克隆抗体,并建立了一种新的间接竞争化学发光酶免疫分析法(CLEIA)。首先将斯坦诺唑尔半抗原与蛋白偶联制备免疫原和包被抗原。为了提高该方法的性能,对化学发光检测系统进行了优化。研究了4-碘酚、过氧化氢和鲁米诺的最佳浓度和最佳检测时间。在最佳条件下,化学发光免疫分析法在磷酸盐缓冲盐水(PBS)中的ic50值为0.34,检测限为0.07 ng/mL。并将该方法与ELISA和HPLC的准确度进行了比较。采用不同的营养药品、食品和生物基质加标斯坦诺唑尔,研究了该方法的应用,平均回收率为92 ~ 113%,变异系数为8 ~ 13.5%。结果表明,基于多克隆抗体的化学发光免疫分析法是一种可行、可靠的斯坦诺唑尔定量方法。
Stanozolol is one of the most widely used synthetic anabolic steroids but also presents a host of negative side effects. It is necessary to develop a rapid and sensitive method for stanozolol screening to monitor its abuse. In this study, the polyclonal antibody against stanozolol was prepared and a novel indirect competitive chemiluminescence enzyme immunoassay (CLEIA) for stanozolol was developed. The stanozolol hapten was conjugated to proteins to prepare immunogen and coating antigens, firstly. To improve the performances of the method, the chemiluminescent detection system was optimized. The optimal concentration of 4-iodophenol, hydrogen peroxide and luminol was studied and the proper detection time was also investigated. Under the optimal conditions, the IC50value and the limit of detection (IC20) of the chemiluminescence immunoassay is 0.34 and 0.07 ng/mL in phosphate-buffered saline (PBS), respectively. The accuracy performances of the proposed CLEIA were compared with that of ELISA and HPLC. Different nutraceuticals, foodstuffs, and biological matrices spiked with stanozolol were applied to study the application of the proposed assays, and the average recoveries locate at 92–113 % with the coefficients of variation of 8–13.5 %. The polyclonal antibody-based chemiluminescence immunoassay formats are proved to be a feasible and reliable quantificational method for stanozolol.
DOI: 10.1109/icbbe.2009.5162231
发表时间: 2009-06
期刊: 2009 3rd International Conference on Bioinformatics and Biomedical Engineering
影响因子: --
作者:
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发表时间: 2009-09
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DOI: 10.1016/s0021-9673(02)01801-0
发表时间: 2003-01
期刊: Journal of chromatography. A
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作者:
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DOI: 10.1016/s0003-2670(02)00672-4
发表时间: 2002-11-25
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通讯作者: Van Peteghem, C
DOI: 10.1016/s0076-6879(72)25042-x
发表时间: 1972-01-01
影响因子: --
作者:
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