An Entire Process Optimization Strategy for Comprehensive In Vivo Metabolite Profiling of Prucalopride in Rats Based on Ultra-Performance Liquid Chromatography With Q-Exactive Hybrid Quadrupole-Orbitrap High-Resolution Mass Spectrometry

Lihua Zuo; Liwei Liu; Yantao Yang; Jie Yang; Min Chen; Huafeng Zhang; Jian Kang; Xiaojian Zhang; Jiabo Wang; Zhi Sun

doi:10.3389/fphar.2021.610226

An Entire Process Optimization Strategy for Comprehensive In Vivo Metabolite Profiling of Prucalopride in Rats Based on Ultra-Performance Liquid Chromatography With Q-Exactive Hybrid Quadrupole-Orbitrap High-Resolution Mass Spectrometry

Front Pharmacol. 2021 May 7:12:610226. doi: 10.3389/fphar.2021.610226. eCollection 2021.

Authors

Lihua Zuo^{1

2}, Liwei Liu^{1

2}, Yantao Yang^{1

2}, Jie Yang^{1

2}, Min Chen³, Huafeng Zhang⁴, Jian Kang^{1

2}, Xiaojian Zhang^{1

2}, Jiabo Wang⁵, Zhi Sun^{1

2}

Affiliations

¹ Department of Pharmacy, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
² Henan Engineering Research Center of Clinical Mass Spectrometry for Precision Medicine, Zhengzhou, China.
³ Department of Neurology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
⁴ Department of Orthopedics, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
⁵ School of Traditional Chinese Medicine, Capital Medical University, Beijing, China.

Abstract

Prucalopride was widely used for chronic constipation, which is difficult to be adequately relieved by laxatives in adult patients in clinic. Due to the difficulty in metabolite identification, metabolic process of prucalopride had not been investigated in vivo. In this study, an efficient strategy was proposed for comprehensive metabolite profiling of prucalopride after oral administration in rat plasma, urine, and feces samples. This strategy was composed of five steps. First, the samples at multiple time points after oral administration were collected to increase the representativeness of the samples. Second, different sample preparation methods were investigated to obtain superior extraction efficiency. Third, the raw data of test sample and blank sample were acquired using ultra-performance liquid chromatography with Q-Exactive hybrid quadrupole-orbitrap high-resolution accurate mass spectrometry under the positive and negative full-scan/dd MS² mode. Fourth, combined mass defect filter with background subtraction model in soft of compound discovery, all peaks were constructed to filter potential metabolites after retention time alignment and ion filtration, which could remove large amounts of interference ions. Besides, it can predict potential biotransformation, promoting to understand how to metabolize the drug. This provides multiple possibilities and prevents us conjecturing the potential metabolites blindly. Finally, the verification procedure was implemented through exporting the structure and MS² spectrum to the analytical tool of Mass Frontier. The proposed strategy significantly improved the targeted detection and identification for metabolites in vivo. A total of 47 metabolites were tentatively characterized in the plasma, urine, and feces samples after oral administration of prucalopride. This study could provide a valuable reference for systematic metabolite profile of drug in vivo.

Keywords: high-resolution mass spectrometry; metabolic network; metabolite identification; prucalopride; the entire and novel strategy.