Zhou Hongyan, Song Shuyi, Lan Xianming, Li Yanan, Yuan Xiaoqing, Yang Jingyi, Li Min, Cao Ting, Zhang Jiayu
School of Pharmacy, Binzhou Medical University, Yantai, Shandong 264003, China.
School of Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan, Shandong 250300, China.
ACS Omega. 2023 Mar 9;8(11):9934-9946. doi: 10.1021/acsomega.2c07089. eCollection 2023 Mar 21.
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 MS 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.
芒果苷是一种具有多种生物活性（如抗炎、抗氧化、抗糖尿病和抗肿瘤）的天然黄酮类化合物，近年来受到了越来越多的研究关注。然而，芒果苷的代谢过程尚未得到充分研究。在本研究中，一种名为“药物代谢物簇”的快速高效分析策略被应用于口服给药后大鼠体内血浆、尿液和粪便样本以及体外肝微粒体中芒果苷代谢物的全面分析。首先，生物样本用甲醇、乙腈和固相萃取（SPE）进行预处理，以进行进一步的液相色谱 - 质谱（LC - MS）分析。其次，在正、负离子全扫描/dd MS模式下，使用超高效液相色谱四极杆精确轨道阱高分辨率质谱（UHPLC - Q - Exactive Orbitrap HRMS）采集原始数据。第三，参考相关文献，选择芒果苷及其基本代谢物（去甲氧基矢车菊素、三羟基氧杂蒽酮和二羟基氧杂蒽酮）作为芒果苷代谢物簇中心。随后，根据质谱裂解规律、文献报道和对照品比较，特别是诊断产物离子（DPI），对候选代谢物进行准确的初步鉴定，并形成基于代谢物簇中心变化的芒果苷代谢物簇。结果，共检测到67种芒果苷代谢物（包括芒果苷），其中血浆中29种，尿液中48种，粪便中12种，肝微粒体中6种。其中，三羟基氧杂蒽酮首次在口服芒果苷后的大鼠尿液样本中被检测到。我们发现芒果苷在大鼠体内主要经历了去糖基化、去羟基化、甲基化、葡萄糖醛酸化、硫酸化等复合反应。在此，我们阐明了芒果苷在体内和体外的代谢物及代谢途径，这为芒果苷的进一步药理研究提供了重要的理论基础，也为药物代谢物的鉴定提供了一种全面的研究方法。
