STUDIES THIAMINE DISULFIDE:III. DETERMINATION OF O-BENZOYLTHIAMINE DISULFIDE

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Reducting conditions of BTDS by cysteine and thiosulfate were investigated and the basal assay procedure was established.1. The optimum condition for reduction with cysteine is the use of 5mg of cysteine hydrochloride for 1 to 100μg BTDS at pH 8.4 to 8.8 and 37° for 60 minutes. The lower the pH of the solution, the more cysteine is required. At pH 7.0 more than 10mg of cysteine is necessary.2. O-Benzoylthiamine disulfide (BTDS) is converted into O-benzoylthiamine and thiamine by cysteine reduction, while O-benzoylthiamine alone is produced by thiosulfate reduction. O-Benzoylthiamine thus formed is converted into thlochrome by alkaline oxidation with cyanogen bromide. The fluorescence intensity produced from thiamine was practically the same as that from O-benzoylthiamine.3. The optimum condition for reduction with thiosulfate is the use of 40mg of sodium thiosulfate for 1 to 100μg of BTDS at pH 4.0 to 5.0, heating at 60° for 20 minutes.4. Adsorbability of O-benzoylthiamine to permutit is poor. However, it become satisfactory after Takadiastase treatment.5. The thiamine formed from BTDS by reduction and Takadiastase treatment is well adsorbed on permutit but has a tendency to be eluted less. 25ml of the eluting solution is not enough but 40ml is sufficient for the recovery more than 95 per cent.
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