Development and validation of an UPLC-UV method for determination of a novel triazine coccidiostat ethanamizuril and its metabolite M3 in chicken tissues

- Ethanamizuril (EZL) is a novel anti-coccidial triazine research and developed in our laboratory.
- Metabolite (M3) was one of EZL's metabolites which mainly was metabolized in kidney.
- We developed a UPLC-UV method for measurements of EZL and M3 in chicken tissue samples.
- Recoveries ranging from 87.3% to 103.2% between all chicken tissue samples were obtained.

Ethanamizuril (EZL) is a novel anti-coccidial triazine synthesized by the Shanghai Veterinary Research Institute of the Chinese Academy of Agricultural Sciences. As a novel coccidiostat, it is essential to study its residue in food animal tissues. In this study, a reliable UPLC method for the determination of ethanamizuril (EZL) and one of its metabolites (M3) in chicken muscle, skin and fat, liver, and kidney samples was developed. Analytes were extracted using acetonitrile and 10% sodium carbonate solution, defatted by n-hexane, and further purified by using an Oasis MCX cartridge. The calibration curves for both test compounds were linear in all tissue matrices we tested: muscle, skin and fat, liver and kidney. The mean relative recoveries of EZL and M3 were from 87.3% to 103.2% between all tissue samples. The inter-day and intra-day relative standard deviations were within 15% and 20%, respectively. This method is accurate, reproducible and is ready to be field-tested.