Wang Bo, Nam Soyeon, Kim Eunyeong, Jeon Hayoung, Lee Kiho, Xie Kaizhou
College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China.
College of Pharmacy, Korea University, Sejong 30019, Korea.
Foods. 2021 Jun 29;10(7):1504. doi: 10.3390/foods10071504.
Nontargeted analysis can be used for the rapid screening and confirmatory analysis of veterinary drugs and their metabolites, which are important for the comprehensive safety evaluation of animal-derived foods. Here, a novel nontargeted screening approach based on liquid chromatography coupled with electrospray ionization-high-resolution mass spectrometry (LC/ESI-HR-MS) was developed to determine erythromycin, clarithromycin, and their metabolites in chicken liver microsomes. Erythromycin and clarithromycin were incubated in vitro in the presence of NADPH for 60 min to generate metabolites in chicken liver microsomes. After the incubation, the supernatant was extracted using ultrasonic shaking, orbital shaking, and centrifugation before analysis using LC/ESI-HR-MS in positive ion mode on an Agilent Eclipse Plus C18 column (100 mm × 2.1 mm; i.d. 3.5 µm) with 0.1 percent formic acid-water and acetonitrile as the mobile phases for gradient elution at 0.4 mL/min. The results show that erythromycin can produce N-desmethyl-erythromycin A in chicken liver microsomes, but clarithromycin cannot produce N-desmethyl-clarithromycin in chicken liver microsomes. The N-desmethyl-erythromycin A and N-desmethyl-clarithromycin were tentatively identified in chicken liver microsomes using the established quick analytic method, which will provide a theoretical foundation for future research on pharmacokinetics and drug elimination in poultry.
非靶向分析可用于兽药及其代谢物的快速筛查和确证分析，这对于动物源性食品的全面安全性评估非常重要。在此，开发了一种基于液相色谱与电喷雾电离-高分辨率质谱联用（LC/ESI-HR-MS）的新型非靶向筛查方法，用于测定鸡肝微粒体中的红霉素、克拉霉素及其代谢物。将红霉素和克拉霉素在NADPH存在下于体外在鸡肝微粒体中孵育60分钟以生成代谢物。孵育后，通过超声振荡、回旋振荡和离心提取上清液，然后在Agilent Eclipse Plus C18柱（100 mm×2.1 mm；内径3.5 µm）上以0.1%甲酸-水和乙腈作为流动相进行梯度洗脱，流速为0.4 mL/min，采用正离子模式通过LC/ESI-HR-MS进行分析。结果表明红霉素在鸡肝微粒体中可产生N-去甲基红霉素A，但克拉霉素在鸡肝微粒体中不能产生N-去甲基克拉霉素。使用已建立的快速分析方法在鸡肝微粒体中初步鉴定出N-去甲基红霉素A和N-去甲基克拉霉素，这将为未来家禽体内药代动力学和药物消除研究提供理论基础。
