Seidler E, van Noorden C J
Institut für Anatomie, Medizinische Fakultät (Charité), Humboldt-Universität Berlin, Germany.
Acta Histochem. 1994 Mar;96(1):43-9. doi: 10.1016/S0065-1281(11)80007-3.
Thin layer chromatography (TLC) was performed to analyze which products are formed when unstable tetrazolinyl radicals are generated during the reduction of tetrazolium salts under physiological conditions of biochemical and histochemical assays. It was found in aqueous media that irrespective the assay conditions, reduction of 2,3-di(4-nitrophenyl)-5-phenyl-2H-tetrazolium chloride (p-DNTTC) or 2,3-di(4-nitrophenyl)-5-t-butyl-2H-tetrazolium chloride (t-butyl-DNTC) always lead to production of formazan and never to generation of the corresponding 2(4-nitrophenyl)-5-phenyl- (respectively t-butyl-) tetrazoles by splitting off a nitrophenyl side chain from the tetrazolium ring as a cation. This reaction is known to occur in non-aqueous media (Neugebauer, 1973). Because p-DNTTC is analogous to tetrazolium salts that are used in biochemical and histochemical assays, it is concluded that reduction of tetrazolium salts leads to formation of formazans only under these conditions.
进行薄层色谱法（TLC）分析，以确定在生化和组织化学分析的生理条件下，四唑盐还原过程中产生不稳定的四唑啉基自由基时会形成哪些产物。在水性介质中发现，无论分析条件如何，2,3-二（4-硝基苯基）-5-苯基-2H-四唑氯化物（p-DNTTC）或2,3-二（4-硝基苯基）-5-叔丁基-2H-四唑氯化物（叔丁基-DNTC）的还原总是导致生成甲臜，而不会通过从四唑环上作为阳离子分裂出硝基苯基侧链而生成相应的2（4-硝基苯基）-5-苯基-（分别为叔丁基-）四唑。已知该反应在非水性介质中发生（Neugebauer，1973）。由于p-DNTTC类似于生化和组织化学分析中使用的四唑盐，因此得出结论，四唑盐的还原仅在这些条件下导致甲臜的形成。
