A High Sensitive Determination of Glutathione and Glutathione Disulfide by HPLC with Post-Label Fluorometric Detection.

Abstract

A high sensitive high-performance liquid chromatographic method for determination of pmol levels glutathione (GSH) and glutathione disulfide (GSSG) in biological samples is described. GSH and GSSG were separated isocratically on a reversed-phase column (ODS column). The mobile phase consisted of 50mM sodium phosphate buffer (pH 3.0). After chromatographic separation, GSH and GSSG were converted to fluorescent derivatives by post-label reaction with o-phthalaldehyde under the alkaline condition. A 0.5pmol quantity of GSH and GSSG can be determined and the analytical recoveries were found to be almost 100%. The procedure is rapid, completed within 15 min, and it has. been successfully applied to various biological samples.