Piccoli G, Fiorani M, Biagiarelli B, Palma F, Potenza L, Amicucci A, Stocchi V
Istituto di Chimica Biologica Giorgio Fornaini, Università di Urbino, Italy.
J Chromatogr A. 1994 Jul 29;676(1):239-46. doi: 10.1016/0021-9673(94)80465-6.
This paper describes a high-performance capillary electrophoretic (HPCE) method which allows a quick, simultaneous and quantitative determination of reduced (GSH) and oxidized (GSSG) glutathione in mammalian red blood cells using a Supelco-bonded hydrophilic phase capillary CElect-P150. The extraction procedure of GSH and GSSG from erythrocytes using Microcon-10 membranes is very simple and allows a correct evaluation of these compounds present in the red blood cells. Furthermore, the HPCE method does not require removal of the excess N-ethylmaleimide used to block the glutathione in its reduced state, making the simultaneous evaluation of GSH and GSSG possible in a very short time (ca. 4 min), with a sensitivity at femtomole level.
本文描述了一种高效毛细管电泳(HPCE)方法,该方法使用Supelco键合亲水性毛细管CElect-P150,能够快速、同时且定量地测定哺乳动物红细胞中还原型谷胱甘肽(GSH)和氧化型谷胱甘肽(GSSG)。使用Microcon-10膜从红细胞中提取GSH和GSSG的过程非常简单,能够正确评估红细胞中这些化合物的含量。此外,HPCE方法无需去除用于阻断还原态谷胱甘肽的过量N-乙基马来酰亚胺,从而能够在极短时间内(约4分钟)同时评估GSH和GSSG,灵敏度可达飞摩尔水平。