Simultaneous determination of glycyrrhizin metabolites formed by the incubation of glycyrrhizin with rat feces by semi-micro high-performance liquid chromatography.
A method for semi-micro high-performance liquid chromatography (HPLC) has been established for the simultaneous determination of 3alpha-hydroxyglycyrrhetic acid and 3-dehydroglycyrrhetic acid together with glycyrrhizin, glycyrrhetic acid and glycyrrhetic acid mono-glucuronide formed by incubation of glycyrrhizin with rat feces. The analysis was accomplished within 25 min with a TSKgel ODS-80TsQA (150 x 2.0 mm i.d.) column by linear gradient elution using a mobile phase containing aqueous phosphoric acid and acetonitrile at a flow rate of 0.2 ml.min(-1), a thermostatic oven at 25 degrees C, and detection at 254 nm. The detection limits of these compounds were 0.2 pmol per injection (5 microl). The metabolites of glycyrrhizin, by anaerobic or aerobic incubation with rat fecal suspension over 48 h, were determined. Glycyrrhizin was almost completely converted to metabolite glycyrrhetic acid, and metabolites 3alpha-hydroxyglycyrrhetic acid and 3-dehydroglycyrrhetic acid in negligible amounts in anaerobic conditions. However, the metabolic time courses of 3-dehydroglycyrrhetic acid when incubated in aerobic conditions revealed that it apparently continued increasing during the whole incubation period.
Okamura
Nobuyuki
N
Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, Gakuen-cho, Hiroshima, Japan. okamura@fupharm.fukuyama-u.ac.jp
Miyauchi
Hidetoshi
H
Choshi
Tominari
T
Ishizu
Takashi
T
Yagi
Akira
A
D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000032:analysis* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D005243:Feces / Q000737:chemistry*; D006034:Glycyrrhetinic Acid / Q000031:analogs & derivatives / Q000032:analysis / Q000378:metabolism; D019695:Glycyrrhizic Acid / Q000032:analysis* / Q000378:metabolism; D008297:Male; D020835:Medicine, Kampo; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
C482687:3-alpha-hydroxyglycyrrhetic acid; D000894:Anti-Inflammatory Agents, Non-Steroidal; C482685:glycyrrhetic acid mono-glucuronide; D019695:Glycyrrhizic Acid; D006034:Glycyrrhetinic Acid