Determination of urinary 18 beta-glycyrrhetinic acid by gas chromatography and its clinical application in man.

A sensitive and quantitative gas chromatographic assay for the determination of 18 beta-glycyrrhetinic acid (18 beta-GA), the main metabolite of glycyrrhizin after oral licorice consumption in human urine, has been developed and validated. For the extraction of 18 beta-GA from urine two Sep-Pak C18 extractions, hydrolysis with Helix pomatia and three liquid-liquid extractions were performed, using 18 alpha-glycyrrhetinic acid (18 alpha-GA) as internal standard. Both 18 beta-GA and internal standard were converted into their pentafluorobenzyl-ester/trimethylsilyl-ether derivatives and detected by flame ionization detection using a WCOT-fused-silica capillary column. Good quality control data were obtained in precision and accuracy tests. The detection limit of the gas chromatographic method was 10 micrograms/l with a urine volume of 10 ml. A detection limit of 3 micrograms/l was obtained by performing GC-MS. The GC method was used to monitor the urinary excretion of 18 beta-GA after licorice consumption by two healthy volunteers and a patient suspected of licorice abuse. Furthermore, it was shown that this GC assay enables to detect other metabolites related to licorice consumption.