Takamitsu Sekine, Kimiyo Ando, Minoru Machida
Aug 1, 1972
Citations
0
Influential Citations
43
Citations
Journal
Analytical biochemistry
Abstract
Abstract 1. 1. N-(p-(2-Benzimidazolyl)phenyl)maleimide (BIPM) has been used for the quantitative determination of thiol compounds. The method is based on the particular fluorescence characteristics of this reagent: its adducts to thiol compounds have strong fluorescence (λmax, ex 315 mμ, λmax, em 365 mμ), while the reagent itself has no appreciable fluorescence. 2. 2. The addition reaction of 2 × 10−6M BIPM to thiols is completed within 30 min at pH 6.85 and 0°C. A linear relation was observed up to 10−5M of the reagent between concentration of adduct and intensity of fluorescence. 3. 3. A very sensitive fluorometric determination of thiol compounds has been developed based on the above results. The detection limit was 10−6M concentration of thiols, with 0.1 ml of sample, corresponding to 0.03 μg reduced glutathione, with an accuracy of 3%. 4. 4. Using the fluorometric method, the oxidation rate of glutathione catalyzed by metal ions was examined and the contents of thiols in human lacrimal fluid and seurm were determined.