An automatic method for the determination of thiamine andits phosphates by the application of immobilized thiaminase I was examined.
Thiaminase I catalyzed the base-exchange reaction of the thiamine phosphates as well as thiamine with pyridine, forming heteropyrithiamine(HPT). The contents of thiamine and its phsphates were determined by automatically analyzing HPT formed by the enzyme reaction. It was considered that this system was suitable for the determination of total thiamine.