A new direct HPLC procedure for the simultaneous determination of sulbutiamine (Arcalion) and other thiamine disulfides in human plasma has been developed. The method involves an automated solidphase extraction on octadecylsilyl (C18) cartridges and chromatographic separation of the compounds on an RP Select B column with gradient elution using methanol and phosphate buffer. Detection was by fluorescence of the resulting thiochromes obtained from two on-line post-column reactors. Optimization of post-column reaction parameters has been achieved. This method has been proved to be highly selective for the determination of the thiamine disulfide derivatives and quantitation limits of 5 ng·ml−1were obtained for each compound in human plasma. Linearity was in the range 5–200 ng·ml−1. Precision and accuracy were also demonstrated by within-day and between-day assays, and showed the good reliability of the