筛查食品中违法添加奥沙普嗪荧光免疫层析方法研究
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摘要
目的本研究建立了一种筛查食品中违法添加的奥沙普嗪胶体金免疫检测试纸条(ICA);方法以改良活泼酯法将奥沙普嗪发表连接到BSA和OVA,合成免疫抗原和检测抗原,免疫家兔获得针对抗奥沙普嗪抗体;以胶金垫、NC膜、样品垫和吸水垫组装IAC试剂条,以标记抗体的40nm胶体金,分别与NC膜上的检测抗原和山羊抗兔IgG二抗进行免疫反应,形成检测线(T线)和质控线(C线)。结果在ELISA体系检测奥沙普嗪的IC_(50)值为13.2ng·mL~(-1),与8种相关药物交叉反应率都小于0.1%,优化IAC试剂条对奥沙普嗪的最低检测限为2.0ng·mL~(-1)。IAC试剂对30个市售样品的检测结果,与HPLC验证结果吻合。结论本方法灵敏、特异、简便、快速,是对违法添加奥沙普嗪的食品进行筛查的有效工具。
OBJECTIVE An immunochromatographic assay( ICA) for screening oxaprozin as illicit adulterant in food was developed in this research. METHODS Artificial antigens were prepared by conjugating oxaprozin to OVA and BSA using modified active ester mechanism. Antibodies against oxaprozin were raised from rabbits by immunizing with BSA-oxaprozin conjugate. RESULTS Results indicated that the IC_(50) value for oxaprozin was 13. 2 ng·mL~(-1) in the ELISA. The cross-reactivities for 8 associate drugs were lower than 0. 1%. A consequent ICA showed detection limit of 2. 0 ng·mL~(-1) for oxaprozin. The current ICA showed good agreements with HPLC in detecting oxaprozin in 30 food samples from local market. CONCLUSION The ICA is suitable for screening illegal adulterant of oxaprozin in foodstuffs due to highly sensitive and specific.
OBJECTIVE An immunochromatographic assay( ICA) for screening oxaprozin as illicit adulterant in food was developed in this research. METHODS Artificial antigens were prepared by conjugating oxaprozin to OVA and BSA using modified active ester mechanism. Antibodies against oxaprozin were raised from rabbits by immunizing with BSA-oxaprozin conjugate. RESULTS Results indicated that the IC_(50) value for oxaprozin was 13. 2 ng·mL~(-1) in the ELISA. The cross-reactivities for 8 associate drugs were lower than 0. 1%. A consequent ICA showed detection limit of 2. 0 ng·mL~(-1) for oxaprozin. The current ICA showed good agreements with HPLC in detecting oxaprozin in 30 food samples from local market. CONCLUSION The ICA is suitable for screening illegal adulterant of oxaprozin in foodstuffs due to highly sensitive and specific.
引文
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[12]王仕平,黄炳泉,刘卿,等.中成药中非法添加奥沙普嗪钠与尼美舒利的快速检测[J].中国药品标准,2011,12(3):211-214.
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[14]Peiwu Li,Qi Zhang,Wen Zhang,et al.Development of a class-specic monoclonal antibody-based ELISA for a atoxins in peanut[J].Food Chemistry,2009,115(2):313-317.
[15]Chunmei Wang,Yihua Liu,Yirong Guo,et al.Development of a Mc Ab-based immunoassay for parathion and inuence of the competitor structure[J].Food Chemistry,2009,115(2):365-370.
[2]宋辉,陈为飞.冯了性风湿跌打药酒假药中奥沙普嗪钠定性、定量分析[J].中国现代应用药学,2002,19(5):405.
[3]苏戈,王培松,王颖.奥沙普嗪钠不良反应[J].中国误诊学杂志,2008,8(16):4014-15.
[4]王文森,李永申,陈敬然.奥沙普嗪钠凝胶引起全身药敏性皮炎一例[J].中国误诊学杂志,2008,7(16):1527-28.
[5]张洁.奥沙普嗪钠致过敏性休克1例[J].医药导报,2007,26(6):648.
[6]宁素云,郭兴杰,张虹,等.HPLC法同时检测清热解毒类中成药中非法添加的9种化学药品[J].中国药事,2009,23(9):1067-69.
[8]赵凤菊,来国防,孙刚,等.UPLC/MS/MS法检测抗风湿类制剂中添加醋酸泼尼松、醋酸地塞米松、奥沙普嗪钠和布洛芬[J].药物分析杂志,2010,30(6):1035-37.
[7]刘福艳,姜俊,谢元超,等.液相色谱-质谱联用法检测中药制剂中掺入的奥沙普嗪、醋酸泼尼松[J].药物分析杂志,2009,29(6):2131-33.
[10]燕朝丽.液质联用检测消肿止痛中成药的氨基比林及奥沙普嗪钠[J].中国药师,2010,13(12):1773-75.
[11]刘晓哲,闻京伟.液质联用检测中成药中添加的双氯酚酸钠和布洛芬[J].药物分析杂志,2011,31(4):779-782.
[12]王仕平,黄炳泉,刘卿,等.中成药中非法添加奥沙普嗪钠与尼美舒利的快速检测[J].中国药品标准,2011,12(3):211-214.
[13]邓树勇,马启龙,丁燕,等.TLC法快速筛选抗风湿类中成药中的化学药品[J].药物分析杂志,2009,29(10):1719-21.
[14]Peiwu Li,Qi Zhang,Wen Zhang,et al.Development of a class-specic monoclonal antibody-based ELISA for a atoxins in peanut[J].Food Chemistry,2009,115(2):313-317.
[15]Chunmei Wang,Yihua Liu,Yirong Guo,et al.Development of a Mc Ab-based immunoassay for parathion and inuence of the competitor structure[J].Food Chemistry,2009,115(2):365-370.