Quantitation of urinary 1,2,3,4-tetrahydro-beta-carboline and 1-methyl-1,2,3,4-tetrahydro-beta-carboline by high-performance liquid chromatography.

A high-performance liquid chromatographic method was developed to quantify 1,2,3,4-tetrahydro-beta-carboline (TBC) and 1-methyl-1,2,3,4-tetrahydro-beta-carboline (MTBC) in human urine. Urine samples with added internal standard were subjected to a reaction with fluorescamine and solvent extractions to remove the precursor tryptamine, which readily condenses with aldehydes in samples and reagents. Such a pretreatment completely suppressed the artifactual formation of TBC and MTBC during analytical procedures. The purified original tetrahydro-beta-carbolines and the internal standard were separated by reversed-phase ion-pair chromatography with fluorescent detection. Their simultaneous separation was automatically completed in a short time (< 12 min). Both TBC and MTBC were quantified at ng/mL concentrations. The quantitative results revealed a wide variation in urinary levels of TBC and MTBC, possibly indicating that their considerable amounts excreted in the urine originate from dietary sources.