Stearns D M, Wetterhahn K E
Department of Chemistry, Burke Laboratory, Dartmouth College, Hanover, New Hamsphire 03755-3564.
Chem Res Toxicol. 1994 Mar-Apr;7(2):219-30. doi: 10.1021/tx00038a016.
Reaction of potassium dichromate with sodium ascorbate was studied by EPR spectroscopy at room temperature, in 0.10 M N-[2-hydroxyethyl]piperazine-N'-[2-ethanesulfonic acid] (HEPES), phosphate, cacodylate, and tris(hydroxymethyl)aminomethane hydrochloride (Tris.HCl) buffers at pH 7.0, in the presence of 0.10 M spin trap [5,5-dimethyl-1-pyrroline 1-oxide or 2-methyl-N-(4-pyridinylmethylene)-2-propanamine N,N'-dioxide]. Chromium(V), ascorbate radical, CO2-, and other carbon-based spin trap-radical adducts were observed. Chromium(V), CO2-, and the carbon-based radicals were observed at low ratios of ascorbate to chromium, and ascorbate radical was observed at high ratios of ascorbate to chromium. The presence of Cr(IV) was detected indirectly by reaction with Mn(II) and a subsequent decrease in the Mn(II) EPR signal. More Cr(IV) was found for the higher reaction ratios of ascorbate to Cr(VI). The only buffer effect observed was a relative decrease of the Cr(V) signal in Tris.HCl vs HEPES, phosphate, and cacodylate buffers, no change in the radical adducts was observed. There was no evidence for reactive oxygen species an intermediates in this reaction. Addition of the singlet oxygen trap 2,2,6,6-tetramethyl-4-piperidone hydrochloride showed no 2,2,6,6-tetramethyl-1-piperidinyloxy radical formation. The Cr(V) species did not react with dioxygen, and dioxygen did not affect the formation of carbon-based radicals. A mechanism consistent with these observations is discussed.
在室温下,于pH 7.0的0.10 M N-[2-羟乙基]哌嗪-N'-[2-乙磺酸](HEPES)、磷酸盐、二甲胂酸盐和三(羟甲基)氨基甲烷盐酸盐(Tris.HCl)缓冲液中,在0.10 M自旋捕获剂[5,5-二甲基-1-吡咯啉1-氧化物或2-甲基-N-(4-吡啶基亚甲基)-2-丙胺N,N'-二氧化物]存在的情况下,通过电子顺磁共振光谱研究了重铬酸钾与抗坏血酸钠的反应。观察到了铬(V)、抗坏血酸自由基、CO2-以及其他碳基自旋捕获剂-自由基加合物。在抗坏血酸与铬的低比例下观察到了铬(V)、CO2-和碳基自由基,在抗坏血酸与铬的高比例下观察到了抗坏血酸自由基。通过与Mn(II)反应以及随后Mn(II)电子顺磁共振信号的降低间接检测到了Cr(IV)的存在。在抗坏血酸与Cr(VI)的反应比例较高时发现了更多的Cr(IV)。观察到的唯一缓冲液效应是Tris.HCl缓冲液中Cr(V)信号相对于HEPES、磷酸盐和二甲胂酸盐缓冲液的相对降低,未观察到自由基加合物的变化。没有证据表明该反应中有活性氧作为中间体。添加单线态氧捕获剂2,2,6,6-四甲基-4-哌啶酮盐酸盐未显示有2,2,6,6-四甲基-1-哌啶氧基自由基形成。Cr(V)物种不与氧气反应,氧气也不影响碳基自由基的形成。讨论了与这些观察结果一致的机制。