Determination of l -glutathione by spot test and spectrophotometric methods based on its interaction with phenazine

• Published in: Analytical methods Vol. 13; no. 34; pp. 3779 - 3784
• Main Authors: Kalyniukova, Alina, Studenyak, Yaroslav, Cziáky, Zoltán, Jekő, József, Balogh, József
• Format: Journal Article
• Language: English
• Published: Cambridge Royal Society of Chemistry 14.09.2021
• Subjects: Color; Correlation coefficient; Correlation coefficients; Fourier analysis; Fourier transforms; Glutathione; Histograms; Image analysis; Image processing; Linearity; Mass spectroscopy; Phenazine; Scientific papers; Spectrophotometry; Thin layer chromatography
• ISSN: 1759-9660; 1759-9679
• DOI: 10.1039/d1ay00581b

This research paper presents simple and quick eco-friendly spot test and spectrophotometric methods for the determination of l -glutathione. The spot test assay is based on the formation of a color complex with phenazine methosulphate and l -glutathione on a thin-layer chromatography plate followed by image analysis using a scanner as a detector. For analysis, the image was converted into red, green, and blue (RGB) histograms. A series of parameters that influenced the color formation were investigated, and under the optimal conditions, a good linearity was observed in the range of 200–1000 μg mL −1 and 249–1000 μg mL −1 of l -glutathione with correlation coefficients of 0.9907 for B and 0.9903 for G channels. For the spectrophotometric method, a good linearity was obtained in the range of 2.1–60 μg mL −1 of l -glutathione concentration with a correlation coefficient of 0.9961. A mechanism of the reaction of l -glutathione with phenazine was proposed and confirmed by Fourier transform infrared and mass spectroscopy.