Specific postcolumn detection method for HPLC assay of homocysteine based on aggregation of fluorosurfactant-capped gold nanoparticles

Abstract

Gold nanoparticles (GNPs) capped with nonionic fluorosurfactant molecules (Zonyl FSN) were synthesized, and with the colloidal solution as a probe reagent, a new postcolumn colorimetric detection method for HPLC assay of homocysteine (Hcy) has been developed. The FSN-capped GNPs exhibited excellent stability in aqueous solutions, even in the presence of high salt. The aggregation of the GNPs could be induced by either Hey or cysteine, resulting in an absorption decrease of the colloidal solution at 525 nm and an absorption increase at longer wavelengths (600-700 nm); however, the GNPs did not respond to other amino acids and biomolecules such as glutathione, cysteinylglycine, and glucose. Under optimal conditions (i.e., high salt, neutral pH, and ∼70 °C), the color change of the GNP solution could almost complete (∼90%) within ∼30 s upon the addition of Hcy. The high selectivity and very fast kinetics of the reaction make it a promising system for HPLC postcolumn detection. The new technique has been employed to determine total Hey levels in human urine and plasma samples, and the results are satisfactory. © 2007 American Chemical Society.