Fig. 6.

Time-dependent changes in the relative amounts of the AM and the AM-GSH conjugate after termination of the metabolic reactions. The AM and the AM-GSH conjugate were formed through incubation at 37°C for 30 min in a reaction mixture containing 1 mg/ml HLMs, 80 μM 2-oxo-clopidogrel, and 1 mM GSH, as described under Materials and Methods. Aliquots of the reaction mixture were analyzed with ESI-LC/MS at designated times, and the amounts of metabolites M3 to M5 were calculated as the ratios of the areas for the metabolites to that for the IS. The area for the IS was integrated from the EIC at m/z 322.0, the areas for M3 to M5 were integrated from the EIC at m/z 356.1, and the area for AM-GSH was integrated from the EIC at m/z 661.0. ●, area ratio for M3; ■, area ratio for M4; ▴, area ratio for M5; ○, area ratio for AM-GSH.