High-performance liquid chromatography assay of cysteine and homocysteine using fluorosurfactant-functionalized gold nanoparticles as postcolumn resonance light scattering reagents

Herein, a new postcolumn resonance light scattering (RLS) detection approach coupled with high-performance liquid chromatography (HPLC) was developed to detect cysteine and homocysteine. In the established system, the fluorosurfactant-capped gold nanoparticles (AuNPs) were first employed as postcolumn RLS reagents. The detection principle was based on the enhancement of RLS intensity of AuNPs upon the addition of cysteine/homocysteine. The RLS signals were detected by a common fluorescence detector at λEX = λEM = 560 nm. The linear ranges for both cysteine and homocysteine were in the range of 5.0–50 μM. The detection limits were 5.9 pmol for cysteine and 12 pmol for homocysteine at a signal-to-noise ratio of 3. HPLC separation and RLS detection conditions were optimized in detail. The applicability of the proposed method has been validated by detecting cysteine and homocysteine in human urine samples. Recoveries from spiked urine samples were 95.0–103.0%.

► Gold nanoparticles were first employed as postcolumn RLS reagents. ► Optimization of all important HPLC-RLS method parameters was discussed in detail. ► Simultaneous determination of cysteine and homocysteine in human urine samples was successfully achieved. ► The developed method has much potential in a wide variety of analytes by tuning ligands of nanoparticles.