Валідація методики кількісного визначення флорфеніколу у зразках м’язів методом імуноферментного аналізу

Abstract

Амфеніколи – група хімічних сполук з антибактеріальною активністю, до складу входять хлорамфенікол (ХАФ), тіамфенікол (ТАФ), флорфенікол (ФФ) та їх похідники. Встановлені валідаційні характеристики щодо визначення залишків флорфеніколу у зразках м’язів, такі як: здатність виявлення (ССβ) становить 5,0 мкг/кг, рівень відсічення – 3,62 мкг/кг. Найменший вміст флорфеніколу, що може бути визначеним – 0,2 мкг/кг.

Introduction. Amphenicols are a group of chemical compounds with antibacterial activity, including chloramphenicol (HAF), thiamphenicol (TAF), florfenicol (FF) and their derivatives. Florfenicol (FF) is a synthetic antimicrobial agent with a broad spectrum of action and is one of the most commonly used drugs in poultry, and was developed specifically for veterinary medicine. Given the wide range of activity of florfenicol, the high therapeutic effect combined with low toxicity makes it important for use in animal husbandry. The known method of enzyme-linked immunosorbent assay (ELISA) differs favorably from other screening methods by high sensitivity, specificity, simplicity and speed of performance, availability and stability of reagents, the ability to computer processing of measurement results and automation of test steps, which provides high test efficiency. The aim of the work. To validate the method of enzyme-linked immunosorbent assay to determine the residues of florfenicol in the samples of months of different species of animals (cattle, pigs, chickens, geese, turkeys, rabbits) and fish. Materials and methods. The research was conducted on the basis of the State Research Institute for Laboratory Diagnostics and Veterinary Sanitary Expertise. The material for the study was a solution of florfenicol with a concentration of 1 mg/liter. Results of research and discussion. It was found that the highest value (highest response) for control samples is 0.24 μg/kg and the lowest value for enriched samples (lowest response) - 3.62 μg/kg. According to the obtained results, none of the answers for the enriched samples coincides with the range of answers for the control samples. It follows that the detection ability (CCβ) for this screening method reduces or decreases 5.0 μg/kg The cut-off rate of this test is 3.62 μg/kg. Conclusions and prospects for further research. Validation characteristics have been established for the determination of florfenicol residues in muscle samples, such as: detection ability (CCβ) is 5.0 μg/kg, cut-off level is 3.62 μg/kg. The lowest content of florfenicol that can be determined is 0.2 μg/kg. The percentage of return for enriched samples of both groups is 93 %, which corresponds to the specified production.