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Assessment of reduced glutathione: Comparison of an optimized fluorometric assay with enzymatic recycling method
Authors: Roušar Tomáš | Kučera Otto | Lotková Halka | Červinková Zuzana
Year: 2012
Type of publication: článek v odborném periodiku
Name of source: Analytical Biochemistry
Publisher name: Academic Press (Elsevier)
Place: London
Page from-to: 236-240
Titles:
Language Name Abstract Keywords
cze Stanovení redukovaného glutathionu: srovnání optimalizované fluorimetrické metody a enzymatické metody Glutathion je důležitý tripeptid, který se podílí na celé řadě procesů. Cílem naší práce byla optimalizace spektrofluorometrického stanovení redukovaného glutathionu založeného na reakci mezi glutathionem a o-ftaldialdehydem. Nalezené parametry metody byly: intraassay CV <3%, interassay CV = 5,1%, recovery = 98–102%, linearita = 0–1000 μM GSH, kalibrace R2 = 1,00. Také jsme vyšetřovali vliv přítomnosti aminokyselin na fluorescenci, ale žádná interference nebyla nalezena. Nakonec jsme srovnali optimalizovanou fluorometrickou metodu s enzymatickým stanovením glutathionu a nalezli jsme velmi silnou míru korelace (r = 0,99, n = 45). Glutathion; stanovení glutathionu; spectrofluorometrická metoda
eng Assessment of reduced glutathione: Comparison of an optimized fluorometric assay with enzymatic recycling method Glutathione is an important tripeptide involved in a variety of cellular processes. The aim of our work was to optimize a spectrofluorometric assay for reduced glutathione based on the reaction between glutathione and o-phthalaldehyde. We found these parameters of the method: CV <3%, interassay CV = 5.1%, recovery = 98–102%, linearity = 0–1000 μM GSH, calibration R2 = 1.00. We also anticipated possible effect of various amino acids on the fluorescence signal, but no interference was found. We compared the optimized fluorometric method with a popular enzymatic recycling glutathione assay and found very strong correlation of results (r = 0.99, n = 45). Glutathione; Glutathione assay; Spectrofluorometric method

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