Comprehensive Profiling of Mangiferin Metabolites In Vivo and In Vitro Based on the "Drug Metabolite Clusters" Analytical Strategy

Hongyan Zhou; Shuyi Song; Xianming Lan; Yanan Li; Xiaoqing Yuan; Jingyi Yang; Min Li; Ting Cao; Jiayu Zhang

doi:10.1021/acsomega.2c07089

Comprehensive Profiling of Mangiferin Metabolites In Vivo and In Vitro Based on the "Drug Metabolite Clusters" Analytical Strategy

ACS Omega. 2023 Mar 9;8(11):9934-9946. doi: 10.1021/acsomega.2c07089. eCollection 2023 Mar 21.

Authors

Hongyan Zhou^{1

2}, Shuyi Song¹, Xianming Lan¹, Yanan Li^{1

2}, Xiaoqing Yuan¹, Jingyi Yang¹, Min Li³, Ting Cao⁴, Jiayu Zhang¹

Affiliations

¹ School of Pharmacy, Binzhou Medical University, Yantai, Shandong 264003, China.
² School of Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan, Shandong 250300, China.
³ Yantai Yuhuangding Hospital, Yantai 264001, Shandong, China.
⁴ Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences, Beijing 100190, China.

Abstract

Mangiferin, a natural flavonoid compound with multiple biological activities (e.g., anti-inflammatory, anti-oxidant, anti-diabetic, and anti-tumor), has gained increased research interest in recent years. Nevertheless, the metabolic processing of mangiferin has not been fully investigated. In this study, a rapid and efficient analytical strategy named "Drug Metabolite Clusters" was applied for comprehensive profiling of mangiferin metabolites in rat plasma, urine, and feces samples in vivo following oral administration and liver microsomes in vitro. First, the biological samples were pretreated with methanol, acetonitrile, and solid phase extraction (SPE) for further liquid chromatography-mass spectrometry (LC-MS) analysis. Second, the raw data were acquired using ultra-high performance liquid chromatography quadrupole exactive orbitrap high-resolution mass spectrometry (UHPLC-Q-Exactive Orbitrap HRMS) under the positive and negative full-scan/dd MS² modes. Third, mangiferin and its basic metabolites (norathyriol, trihydroxyxanthone, and dihydroxyxanthone) were selected as mangiferin metabolite cluster centers by referring to the relevant literature. Subsequently, according to the pyrolysis law of mass spectrometry, literature reports, and reference material comparison, especially the diagnostic product ions (DPIs), the candidate metabolites were accurately preliminarily identified, and mangiferin metabolite clusters based on metabolite cluster center changes were formed. As a result, a total of 67 mangiferin metabolites (mangiferin included) were detected, including 29 in plasma, 48 in urine, 12 in feces, and 6 in liver microsomes. Among them, trihydroxyxanthones were first detected in rat urine samples after oral mangiferin. We found that mangiferin mainly underwent deglucosylation, dehydroxylation, methylation, glucuronidation, sulfation, and other composite reactions in rats. Herein, we have elucidated the metabolites and metabolic pathways of mangiferin in vivo and in vitro, which provided an essential theoretical basis for further pharmacological studies of mangiferin and a comprehensive research method for the identification of drug metabolites.