4.7 Article

Identification of Erythromycin and Clarithromycin Metabolites Formed in Chicken Liver Microsomes Using Liquid Chromatography-High-Resolution Mass Spectrometry

Journal

FOODS
Volume 10, Issue 7, Pages -

Publisher

MDPI
DOI: 10.3390/foods10071504

Keywords

nontargeted screening; LC/ESI-HR-MS; erythromycin; clarithromycin; metabolites; chicken liver microsomes

Funding

  1. Yangzhou University International Academic Exchange Foundation
  2. Yangzhou University Excellent Doctoral Dissertation Foundation
  3. Postgraduate Research and Practice Innovation Program of Jiangsu Province [SJCX20_1360]
  4. National Research Foundation (NRF) - Ministry of Education of Korea [2019R1A6A1A03031807]
  5. China Agriculture Research System of MOF and MARA [CARS-41]
  6. Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD)
  7. Yangzhou University High-End Talent Support Program

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A novel nontargeted screening approach based on LC/ESI-HR-MS was developed to determine erythromycin, clarithromycin, and their metabolites in chicken liver microsomes. The results revealed that erythromycin can produce N-desmethyl-erythromycin A in chicken liver microsomes, while clarithromycin cannot produce N-desmethyl-clarithromycin in chicken liver microsomes.
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 x 2.1 mm; i.d. 3.5 mu m) 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.

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