Quantification of a-Thujone and its Metabolites in Human Urine after Consumption of a Sage Infusion Using Stable Isotope Dilution Assays

The metabolism of the monoterpene α-thujone was investigated in humans after consumption of sage tea, by analyses of its metabolites 2-hydroxythujone, 4-hydroxythujone, and 7-hydroxythujone in urine. For the quantitation of α-thujone and its metabolites, stable isotope dilution assays were developed. Using d ₆-α-thujone as internal standard, we quantified α-thujone by solid phase microextraction GC-MS and the hydroxythujones with d ₆-2-hydroxythujone, d ₆-4-hydroxythujone, and d ₆-7-hydroxythujone after glucuronide/sulfate deconjugation by LC-MS/MS in urine. After the consumption of 575.0 µg α-thujone, the 4-hydroxythujone and 7-hydroxythujone were detectable in the urine of the volunteers under study, after liberation from their glucuronides/sulfates. The 2-Hydroxythujone was not present in any of the volunteer samples above its detection limit. α-Thujone was detectable in a low amount, with a maximum concentration of 94 ng/L for the volunteer with the highest dose of 14.3 µg/kg bw.