水产品中甲砜霉素、氟苯尼考和氟苯尼考胺酶联免疫多残留测定

Multi-residue Detecting of Thiamphenicol、Florfenicol and Florfenicol-amine in aquatic product by ELISA

关键词:  甲砜霉素  氟苯尼考  氟苯尼考胺  多残留  酶联免疫法

Key words:  Thiamphenicol  Florfenicol  Florfenicol-amine  multi-residue  ELISA

摘要:

【研究目的】建立一种甲砜霉素、氟苯尼考和氟苯尼考胺多残留酶联免疫检测方法。【方法】将氟苯尼考胺与载体蛋白相连作为抗原免疫动物，获得抗甲砜霉素等药物的抗体，建立了水产品中甲砜霉素、氟苯尼考和氟苯尼考胺残留的ELISA检测方法。【结果】人工抗原中氟苯尼考胺与蛋白质分子的结合比约为12∶1，血清效价达到1∶2000倍以上，50%抑制浓度(IC50)为3 礸?L-1左右，3种酰胺醇类药物交叉反应率在30%~156%之间，鱼和虾肌肉组织中的检测限分别为6.3 礸?kg-1和6.1 礸?kg-1, 在鱼和虾肌肉组织中三种药物的回收率在40%~120%之间。【结论】该方法检测范围为0.2~125 礸?L-1，可同时测定甲砜霉素、氟苯尼考及其标示残留物氟苯尼考胺等3种化合物残留量。方法灵敏度、精密度和准确度均能满足兽药残留筛选检测要求。

Abstract:

【Objective】Immunoassay for detecting of Thiamphenicol、Florfenicol and Florfenicol-amine multi-residues was developed.【Method】Immunizing animals with Florfenicol-amine protein combination, anti- thiamphenicol and two others antibody was gotten. An ELISA method for detecting thiamphenicol、Florfenicol and Florfenicol-amine residues in aquatic product was developed. 【Result】The conjugate rate was 12 Florfenicol-amine mole to one protein mole in artificial antigen. Antiserum valence was more than 1∶2000 and the IC50 was about 3 礸?L-1. The cross-reactive rate range for 3 kinds of Amphenicols was from 30% to 156%. The limit of detection were 6.3 礸?kg-1and 6.1 礸?kg-1 in fish and shrimp muscle respectively. The recovery rate range was from 40% to 120% in fish and shrimp muscle. 【Conclusion】The measuring range was from 0.2 to 125 礸?L-1. It could determine Thiamphenicol、Florfenicol and Florfenicol’s marker residue Florfenicol-amine at the same time. The sensibility, precision and accuracy of the method could satisfy the need of screening veterinary residues.