Pañak K C, Ruiz O A, Giorgieri S A, Díaz L E
Cátedra de Química Analítica Instrumental, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Argentina.
Electrophoresis. 1996 Oct;17(10):1613-6. doi: 10.1002/elps.1150171021.
A reproducible, rapid procedure for the determination of glutathione in human blood by micellar electrokinetic chromatography has been developed. Whole blood samples were deproteinized with 5% w/v sulfosalicylic acid (final concentration). After centrifugation, the supernatant was directly injected for analysis, without further derivatization. Separations were performed by using an uncoated capillary of 30 cm effective length and 50 microns internal diameter (ID), 50 mM Tris-HCl, 30 mM sodium dodecyl sulfate (SDS), pH 7.00, as running buffer, and 10-20 kV. On-line detection was carried out at 214 nm and a detection limit in the range of femtomoles was achieved. Under the same experimental conditions, we resolved a mixed standard solution containing glutathione in its oxidize and reduced forms, lipoamide and alpha-lipoic acid. The corresponding migration times were reproducible. The present method allows rapid determination of these compounds, which play a critical role in oxidative stress, in cellular defense against injurious agents and whose levels are related to the toxicology and metabolism of several toxins and drugs, such as antineoplastic agents.
已开发出一种通过胶束电动色谱法测定人血中谷胱甘肽的可重现、快速的方法。全血样品用5%（w/v）磺基水杨酸（终浓度）进行脱蛋白处理。离心后，上清液直接进样分析，无需进一步衍生化。使用有效长度为30 cm、内径（ID）为50微米的未涂层毛细管，以50 mM Tris-HCl、30 mM十二烷基硫酸钠（SDS）、pH 7.00作为运行缓冲液，在10 - 20 kV下进行分离。在214 nm处进行在线检测，实现了飞摩尔范围内的检测限。在相同的实验条件下，我们分离了一种含有氧化型和还原型谷胱甘肽、硫辛酰胺和α-硫辛酸的混合标准溶液。相应的迁移时间具有可重复性。本方法可快速测定这些在氧化应激、细胞对损伤因子的防御中起关键作用且其水平与几种毒素和药物（如抗肿瘤药物）的毒理学和代谢相关的化合物。
