Wang Xiaoyang, Zhao Juan, Wei Shuya, Wang Chunmei, Zhang Lifang, Wang Mi, Liu Yingchun, Fei Chenzhong, Xue Feiqun, Zhang Keyu
Key Laboratory of Veterinary Chemical Drugs and Pharmaceutics, Ministry of Agriculture and Rural Affairs, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China.
School of Biological Engineering, Wuhan Polytechnic, Wuhan, China.
Biomed Chromatogr. 2019 Nov;33(11):e4652. doi: 10.1002/bmc.4652. Epub 2019 Aug 20.
Ethanamizuril is a new triazine compound that has the potential to be a novel anticoccidial drug. Toxicological studies in experimental rats were performed to understand the safety profile of ethanamizuril for drug product development. In this study, a novel, selective and accurate ultra-performance liquid chromatography tandem mass spectrometry method has been developed for the determination of ethanamizuril concentrations in rat plasma. With 4-nitro-o-cresol as an internal standard, sample pretreatment involved a one-step extraction with acetonitrile of 100 μL plasma. The detection was carried out by electrospray ionization mass spectrometry in negative ion mode with selected ion recording. The standard curves were linear (r ≥ 0.999) over the concentration range of 0.1-100 μg/mL. The relative standard deviations of intra- and inter-day precisions were less than 8.4 and 8.87%, respectively. The mean extraction recovery of ethanamizuril from rat plasma was 97.68-102.57%. The method was fully validated and successfully applied to monitor plasma concentrations of ethanamizuril in a short-term toxicity study and two-generation reproduction toxicity study. The result of the study confirmed that the elimination of ethanamizuril in rats is slow.
乙胺嘧啶是一种新型三嗪化合物，有潜力成为一种新型抗球虫药。为了解乙胺嘧啶在药品研发中的安全性，对实验大鼠进行了毒理学研究。在本研究中，开发了一种新颖、选择性好且准确的超高效液相色谱串联质谱法，用于测定大鼠血浆中乙胺嘧啶的浓度。以4-硝基邻甲酚为内标，样品预处理包括用乙腈对100 μL血浆进行一步萃取。检测采用电喷雾电离质谱法，在负离子模式下进行选择离子记录。标准曲线在0.1-100 μg/mL浓度范围内呈线性（r≥0.999）。日内和日间精密度的相对标准偏差分别小于8.4%和8.87%。乙胺嘧啶从大鼠血浆中的平均萃取回收率为97.68-102.57%。该方法经过充分验证，并成功应用于短期毒性研究和两代繁殖毒性研究中乙胺嘧啶血浆浓度的监测。研究结果证实，大鼠体内乙胺嘧啶的消除速度较慢。
