FLUOROMETRIC METHOD FOR DETERMINATION OF OXIDIZED AND REDUCED GLUTATHIONE IN TISSUES
FLUOROMETRIC METHOD FOR DETERMINATION OF OXIDIZED AND REDUCED GLUTATHIONE IN TISSUES
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DOI：
10.1016/0003-2697(76)90326-2
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发表时间：
1976-01-01
影响因子：
2.9
通讯作者：
HILF, R
中科院分区：
文献类型：
--
作者：
HISSIN, PJ;HILF, R
建立了以邻苯二醛（OPT）为荧光试剂测定氧化谷胱甘肽（GSSG）和还原谷胱甘肽（GSH）的方法。该方法利用了谷胱甘肽与OPT在pH 8和GSSG与OPT在pH 12下的反应；GSH可以与n -乙基马来酰亚胺络合，以防止GSH对GSSG测量的干扰。方法对谷胱甘肽和谷胱甘肽的加样回收率为91 ~ 110%，对谷胱甘肽有较好的特异性；这些操作似乎都不会影响大鼠组织中谷胱甘肽的含量。谷胱甘肽水平的结果与早期报告一致，但这里估计的GSSG水平高于早期报告的值。讨论了GSSG水平明显较高的原因。
A method for measurement of both oxidized (GSSG) and reduced (GSH) glutathione was developed, with use of o-phthalaldehyde (OPT) as a fluorescent reagent. The method takes advantage of the reaction of GSH with OPT at pH 8 and of GSSG with OPT at pH 12; GSH can be complexed to N-ethylmaleimide to prevent interference of GSH with measurement of GSSG. The method gave recoveries of 91-110% for both GSH and GSSG and was quite specific for glutathione; none of the manipulations appeared to influence the amount of glutathione present in the rat tissue. Results for GSH levels agreed well with earlier reports but levels of GSSG estimated here were higher than earlier reported values. The reasons for the apparently higher levels of GSSG are discussed.