Thiamine triphosphate (ThTP) was chemically synthesized in 1948 at a time when the only organic triphosphate known was ATP. The first claim of the existence of ThTP in living organisms was made in rat liver, followed by baker’s yeast. Its presence was later confirmed in rat tissues and in plants germs, but not in seeds, where thiamine was essentially unphosphorylated. In all those studies, ThTP was separated from other thiamine derivatives using a paper chromatographic method, followed by oxidation in fluorescent thiochrome compounds with ferricyanide in alkaline solution. This method is at best semi-quantitative, and the development of liquid chromatographic methods suggested that ThTP represents far less than 10% of total thiamine in animal tissues.