Selective and sensitive detection of L-Cysteine via fluorometric assay using gold nanoparticles and Rhodamine B in aqueous medium

Abstract

An assay method based on fluorometric and colorimetric change was developed for selective sensing of important thiol containing amino acid L-Cysteine (L-Cys) by using gold nanoparticles (Au NPs) and Rhodamine B (RhB) in aqueous environment. This fluorometric assay is basically relies on the competitive binding between RhB and Au NPs via electrostatic interaction as well as strong thiol(-SH)-Au NPs bonding via chemisorption. Citrate stabilized Au NPs (diameter ∼27 nm) was synthesized by soft chemical method and characterized by UV–Vis absorption spectroscopy and Transmission electron microscopic (TEM) techniques. The change in non-radiative energy transfer between RhB and Au NPs are responsible for the observed drastic fluorescence quenching of RhB via FRET process. The recovery of fluorescence from the assay solution of RhB/Au NPs after addition of L-Cys was found linear over the concentration range 0.01 μM–1000 μM with experimental limit of detection (LOD) of 0.01 μM. The selective interaction of L-Cys with the mixed solution of RhB/Au NPs was reflected by the color change from wine to bluish-black of the final solution. The proposed fluorometric assay method accompanied with the observed colorimetric change could successfully differentiate other interfering amino acids including thiol (-SH) containing compounds namely L-Methionine, L-Homocysteine and antioxidant glutathione with the high degree of accuracy. Also the LOD is comparable to the concentration of L-Cys present in the blood plasma. The proposed sensing mechanism is also confirmed with pure human urine sample to detect the response of L-Cys in the urine. Therefore, this fluorometric assay technique for detection of L-Cys has great potential with the diagnostic impact for biomedical interest.