Figure 1

Reversed phase separation of thiochrome derivatives of thiamine and thiamine phosphate esters. Panel A. A mixture of derivatized standards (50 pmol each) was injected on a C18 amide column as described in Materials and Methods. The vitamins eluted in the following order: TPP (6.4 min), TMP (7.2 min) and thiamine (10.4 min). Panel B. Chromatogram of the thiochromes generated from a peroxisome enriched fraction (3.1 mg protein/ml) obtained from the Nycodenz gradient (black line), documenting the presence of TPP. Some peaks apparently originate from Nycodenz, as revealed by comparison with the profiles of the corresponding blank Nycodenz fractions, both derivatized (red line) and not derivatized (blue line).