Metabolites identification of glycycoumarin, a major bioactive coumarin from licorice in rats

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 98; pp. 287 - 295
• Main Authors: Wang, Qi, Qiao, Xue, Liu, Chun-fang, Ji, Shuai, Feng, Lin-min, Qian, Yi, Guo, De-an, Ye, Min
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 01.09.2014
• Subjects: Animals; Chromatography, Liquid - methods; Coumarins - chemistry; Coumarins - metabolism; Glucuronidase - metabolism; Glucuronides - chemistry; Glucuronides - metabolism; Glycycoumarin; Glycyrrhiza - chemistry; Glycyrrhiza - metabolism; Glycyrrhiza uralensis; Glycyrrhiza uralensis - chemistry; Glycyrrhiza uralensis - metabolism; Hydroxylation; LC/MS; Male; Mass Spectrometry - methods; Metabolites identification; Microsomes, Liver - metabolism; NMR; Rats; Rats, Sprague-Dawley; LC/MS; Glycycoumarin; Glycyrrhiza uralensis; NMR; Metabolites identification
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2014.06.001

•We studied the metabolism of glycycoumarin, a bioactive compound of licorice.•4 and 10 metabolites were characterized in rats plasma and urine by LC/MS.•Two hydroxylated metabolites were obtained by microbial transformation.•The hydroxylated metabolites were proved to be catalyzed by P450 enzymes.•GCM mainly undergoes isoprenyl hydroxylation and glucuronidation in rats. Glycycoumarin is a major bioactive coumarin of licorice (Glycyrrhiza uralensis), one of the most popular herbal medicines worldwide. In this work, the metabolism of glycycoumarin in rats was investigated. After oral administration of 40mg/kg glycycoumarin, 4 and 10 metabolites were respectively detected in rats plasma and urine samples by liquid chromatography coupled with mass spectrometry (LC/MS). These metabolites were tentatively characterized by analyzing their tandem mass spectra and high-resolution mass spectra, and the structures of glucuronides were confirmed by β-glucuronidase hydrolysis. Glycycoumarin mainly undertakes hydroxylation and glucuronidation metabolism, accompanied by hydrogenation and dehydrogenation as minor reactions. Two hydroxylated metabolites, 4′′-hydroxyl glycycoumarin and 5′′-hydroxyl glycycoumarin, were obtained by microbial transformation of Syncephalastrum racemosum AS 3.264, and their structures were fully identified by 1D and 2D NMR. Both metabolites are new compounds. Furthermore, they were proved to be catalyzed by P450 enzymes by rat liver microsomes incubation experiments. Finally, a metabolic pathway of glycycoumarin in rats was proposed. This is the first systematic study on metabolites identification of glycycoumarin.