Abstract:

【Objective】 Immunoassay kit for detecting of tetracyclines multi-residues was developed. 【Method】 Anti-tetracyclines antibody was gotten by immunizing animals with tetracycline-protein combination. An ELISA method for detecting tetracyclines residues in animal tissues was developed. This method was compared with R-biopharm Company Immunoassay Kit and LC-MS/MS method. 【Result】 The conjugate rate was 15 tetracycline mole to one protein mole in artificial antigen. Antiserum valence was more than 1﹕2 000 and the IC50 was about 3 μg?L-1. The cross-reactive rate range for 7 kinds of tetracyclines was from 58% to 100%. The limit of detection were 6.6 μg?L-1and 6.5 μg?kg-1 in milk and chicken muscle, respectively. The recovery rate range was from 40% to 120% in chicken muscle. 【Conclusion】 The lowest detection limit of this test kit was 0.3 μg?L-1 and the measuring range was from 0.3 to 30 μg?L-1. It can determine tetracycline, chlortetracycline, oxytetracycline and deoxycycline usually used in veterinary clinic at the same time. The sensibility, precision and accuracy of the method can satisfy the need of detecting veterinary residue.

Key words: tetracyclines, multi-residue, ELISA, test kit