ENZYMATIC-SYNTHESIS, OPTICAL CHARACTERISTICS, AND QUANTITATIVE-DETERMINATION OF DIHYDROXYETHYLTHIAMINE PYROPHOSPHATE

Dihydroxyethylthiamine pyrophosphate (DHETP), an intermediate of the t ransketolase reaction, was synthesized and purified by a modification of methods reported previously by Holzers group. The absorption spectr um, the CD spectrum, and the value of the molar CD absorption coeffici ent (at 273 nm) of the intermediate are presented. A rapid and conveni ent quantitative assay for DHETP is described, allowing its quantitati ve determination in solutions (whether it is present as a single compo nent or as a mixture with thiamine pyrophosphate and other optically n eutral compounds). A method for quantitative determination of thiamine pyrophosphate in the presence of DHETP is also proposed.