通过毛细管区带电泳同时分析还原型谷胱甘肽和氧化型谷胱甘肽。
Simultaneous analysis of reduced glutathione and glutathione disulfide by capillary zone electrophoresis.
作者信息
Hempe James M, Ory-Ascani Jeannine
机构信息
Research Institute for Children, Children's Hospital, New Orleans, LA, USA; Department of Pediatrics, Louisiana State University Health Sciences Center, New Orleans, LA, USA.
出版信息
Electrophoresis. 2014 Apr;35(7):967-71. doi: 10.1002/elps.201300450. Epub 2014 Feb 8.
This report describes modifications to a CZE method developed by Serru et al. (Clinical Chemistry 2001, 47, 1321-1324) for the simultaneous analysis of reduced glutathione (GSH) and glutathione disulfide (GSSG). Lowering the pH of the run buffer (75 mmol/L boric acid, 25 mmol/L bis-Tris) from pH 8.4 to 7.8 markedly improved GSH peak area reproducibility and allowed multiple samples to be analyzed without changing run buffers due to ion depletion. Sample preparation using red blood cells (RBC) instead of whole blood, combined with glutathione extraction at a lower concentration of metaphosphoric acid (5%), increased assay sensitivity and decreased interference. CZE assay results for clinical samples containing 1000 to 3200 μmol GSH/L RBC and 100 to 400 μmol GSSG/L RBC were highly correlated (r(2) ≥ 0.95) with results obtained using a commercial dithionitrobenze-based glutathione assay. The modified CZE assay has proven useful for the analysis of glutathione in both mouse and human RBC.
本报告描述了对Serru等人（《临床化学》，2001年，47卷，1321 - 1324页）开发的一种毛细管区带电泳（CZE）方法的改进，用于同时分析还原型谷胱甘肽（GSH）和谷胱甘肽二硫化物（GSSG）。将运行缓冲液（75 mmol/L硼酸，25 mmol/L双三羟甲基氨基甲烷）的pH从8.4降至7.8，显著提高了GSH峰面积的重现性，并允许在不由于离子耗尽而更换运行缓冲液的情况下分析多个样品。使用红细胞（RBC）而非全血进行样品制备，同时在较低浓度的偏磷酸（5%）下进行谷胱甘肽提取，提高了检测灵敏度并减少了干扰。对于含有1000至3200 μmol GSH/L RBC和100至400 μmol GSSG/L RBC的临床样品，CZE检测结果与使用基于二硫代硝基苯的商业谷胱甘肽检测方法获得的结果高度相关（r(2)≥0.95）。改良的CZE检测方法已被证明可用于分析小鼠和人类红细胞中的谷胱甘肽。
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