Department of Pharmacy, The First Affiliated Hospital of Kangda College of Nanjing Medical University, The First People's Hospital of Lianyungang, Lianyungang, Jiangsu Province, China.
Department of Clinical Pharmacy, Jingjiang People's Hospital, The Seventh Affiliated Hospital of Yangzhou University, Jingjiang, Jiangsu Province, China.
Rapid Commun Mass Spectrom. 2022 Jun 30;36(12):e9311. doi: 10.1002/rcm.9311.
Ophiopogonanone A (OPA) is one of the representative homoisoflavonoids isolated from Ophiopogonis Radix. The aim of this study was to identify and characterize the metabolites of OPA generated in the liver microsomes and hepatocytes of rats and humans.
The metabolites were generated by incubating OPA (5 μM) with liver microsomes or hepatocytes at 37°C. To trap the reactive metabolites, glutathione (GSH, 5mM) was added into microsomal incubations. The metabolite identification and profiling were performed using ultra-high-performance liquid chromatography combined with photo-diode array detector and quadrupole time-of-flight tandem mass spectrometry (LC-Q/TOF-MS). The acquired mass data were processed by MetaboLynx software. The structures of the metabolites were tentatively characterized in terms of their accurate masses, product ions, and retention times.
Under the present conditions, a total of nine metabolites were detected and their structures were tentatively identified. Among these metabolites, M8 (OPA catechol) was the most abundant metabolite both in rat and human liver microsomes. M7 (glucuronidation product of M8) was the major metabolite both in rat and human hepatocytes. The metabolic pathways of OPA include demethylenation, dehydrogenation, hydroxylation, methylation and glucuronidation and GSH conjugation.
Our results provided valuable information regarding the in vitro metabolism of OPA, which would help us understand the mechanism of the elimination of OPA and in turn the effectiveness and potential toxicity.
川麦冬酮 A（OPA）是从麦冬中分离得到的代表性异黄酮之一。本研究旨在鉴定和表征大鼠和人肝微粒体和肝细胞中生成的 OPA 的代谢物。
将 OPA（5 μM）在 37°C 下与肝微粒体或肝细胞孵育以生成代谢物。为了捕获反应性代谢物，将谷胱甘肽（GSH，5mM）加入到微粒体孵育中。使用超高效液相色谱法结合光电二极管阵列检测器和四极杆飞行时间串联质谱法（LC-Q/TOF-MS）进行代谢产物的鉴定和分析。使用 MetaboLynx 软件处理获得的质谱数据。根据准确质量、产物离子和保留时间，初步推断代谢物的结构。
在本研究条件下，共检测到 9 种代谢物，并初步推断其结构。在这些代谢物中，M8（OPA 儿茶酚）是大鼠和人肝微粒体中最丰富的代谢物。M7（M8 的葡萄糖醛酸化产物）是大鼠和人肝细胞中的主要代谢物。OPA 的代谢途径包括去甲基化、脱氢、羟化、甲基化和葡萄糖醛酸化以及 GSH 缀合。
本研究结果提供了 OPA 体外代谢的有价值信息，有助于我们了解 OPA 消除的机制，从而了解其有效性和潜在毒性。
