Development of radioactive tracing coupled with LC/MS-IT-TOF methodology for the discovery and identification of diaveridine metabolites in pigs

Liye Wang; Lihua Wen; Yuanhu Pan; Zhenli Liu; Chenyun Zhang; Zonghui Yuan; Lingli Huang

doi:10.1016/j.foodchem.2021.130200

Development of radioactive tracing coupled with LC/MS-IT-TOF methodology for the discovery and identification of diaveridine metabolites in pigs

Food Chem. 2021 Nov 30:363:130200. doi: 10.1016/j.foodchem.2021.130200. Epub 2021 May 25.

Authors

Liye Wang¹, Lihua Wen², Yuanhu Pan³, Zhenli Liu³, Chenyun Zhang⁴, Zonghui Yuan³, Lingli Huang⁵

Affiliations

¹ National Reference Laboratory of Veterinary Drug Residues (HZAU), Huazhong Agricultural University, Wuhan, Hubei 430070, PR China; College of Food and Drug, Luoyang Normal University, Luoyang, Henan 471934, PR China.
² National Reference Laboratory of Veterinary Drug Residues (HZAU), Huazhong Agricultural University, Wuhan, Hubei 430070, PR China.
³ National Reference Laboratory of Veterinary Drug Residues (HZAU), Huazhong Agricultural University, Wuhan, Hubei 430070, PR China; MAO Key Laboratory for Detection of Veterinary Drug Residues, Huazhong Agricultural University, Wuhan, Hubei 430070, PR China; MOA Laboratory of Risk Assessment for Quality and Safety of Livestock and Poultry Products, Huazhong Agricultural University, Wuhan, Hubei 430070, PR China.
⁴ College of Food and Drug, Luoyang Normal University, Luoyang, Henan 471934, PR China.
⁵ National Reference Laboratory of Veterinary Drug Residues (HZAU), Huazhong Agricultural University, Wuhan, Hubei 430070, PR China; MAO Key Laboratory for Detection of Veterinary Drug Residues, Huazhong Agricultural University, Wuhan, Hubei 430070, PR China; MOA Laboratory of Risk Assessment for Quality and Safety of Livestock and Poultry Products, Huazhong Agricultural University, Wuhan, Hubei 430070, PR China. Electronic address: huanglingli@mail.hzau.edu.cn.

PMID: 34120054
DOI: 10.1016/j.foodchem.2021.130200

Abstract

We developed a sensitive and reliable method by coupling radiotracing with LC/MS-IT-TOF to identify diaveridine metabolites. Tritium-labeled diaveridine was orally administered to pigs and their organs, blood, bile, and excreta were collected. Under optimized conditions, radioactive recovery was >90% and the highest numbers of metabolites were detected. MCX-based solid-phase extraction was conducted for urine, plasma, and bile purification. Methanol-chloroform 1:1 (v/v), methanol-chloroform 6:1 (v/v), methanol, methanol-chloroform 1:1 (v/v), and methanol were used as solvents to extract feces, liver, kidney, fat and muscle, respectively. The method validation confirmed satisfactory ³H-H exchange efficiency (<5%), chromatographic column efficiency (≥97.5%), LOQ (10.73 μg/kg), and analytical accuracy (97.6-107.8%) and precision (RSD < 5%). Moreover, novel in vivo metabolites were detected in the pigs, including D2 (3'-desmethyl-diaveridine monoglucuronide), D3 (diaveridine monoglucuronide). Hence, the analytical method developed herein lays an empirical foundation for further systematic studies of the diaveridine metabolism.

Keywords: Diaveridine; Isotopic tracing; Metabolite; Sample preparation.

MeSH terms

Animals
Chromatography, High Pressure Liquid
Chromatography, Liquid
Feces
Pyrimidines
Solid Phase Extraction*
Swine
Tandem Mass Spectrometry*

Substances

Pyrimidines
diaveridine