Method of Cysteine/Cystine Ratio Assessment Suitable for Physiological Applications

A fluorimetric method for the determination of cysteine (CSH) and cystine (CSSC) at nanomolar concentrations is reported. The detection limit was 2.46x10 -11 M for CSH when derivatisation is performed by 5-iodoacetamidofluorescein (5-IAF) and respectively 7.2x10 -10 M when monobromobimane was used as fluorescent reagent. The dynamic range of analytical response was 3x10 -10 - 3x10 -8 M (in the case of CSH-5-IAF). The method was further applied to determine the (CSH)/(CSSC) ratio at nanomolar concentration levels as potential marker of aminoacids oxidative status. An original algorithm to calculate the exact concentrations for each form was established, being provided even the experimental data to support it. Zinc influence on avoiding CSH oxidation was studied. Data were confirmed by high-performance liquid chromatography with fluorescence detection.

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