Thiamine EP Impurity D (CAS 490-82-4) is a defined pharmacopoeial related substance of thiamine, with molecular formula C12H16N4O2S and molecular weight 280.3 g/mol. Its structural identity reflects a thiamine-derived heteroaromatic degradation product rather than a nitrosamine or classical genotoxic impurity. Formation is typically associated with oxidative or hydrolytic transformation of the thiazolium-containing vitamin scaffold during synthesis, storage, or stress conditions. Because trace levels may persist in complex matrices, characterization commonly relies on LC-MS/MS, high-resolution LC-MS, and orthogonal HPLC-UV methods, with spectral confirmation used to differentiate it from closely related thiamine analogues and process-related impurities.
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