Description
For the determination of glutathione the sample is divided in two aliquots. One is reduced and the total amount of glutathione is measured. The other aliquot is treated without reduction solution, which determines only the reduced glutathione. During the derivatisation reaction glutathione is converted into a fluorescent probe. The following precipitation step removes high molecular substances. After centrifugation the fluorescent probe is cooled (2-8°C) and injected into the HPLC system. The isocratic separation via HPLC at 30°C uses a reversed phase column in two runs. One run lasts 4 minutes. The chromatograms are recorded by a fluorescence detector. The quantification is performed with the delivered EDTA-blood calibrator; the concentration is calculated by the internal standard method