Single-step controlled synthesis of flower-like gold nanoparticles stabilized by chitosan for sensitive detection of heparin using a surface-enhanced Raman scattering method.

A novel single-step and template-free procedure, including controlled synthesis of gold flowers (AuNFs), conjugation to a 4-MBA reporter, and stabilization with chitosan, is proposed to develop the SERS tags-based nanoparticles for trace detection of heparin. This SERS detection assay is based on the aggregation/non-aggregation balance of AuNFs-4-MBA@chitosan nanoparticles, which was induced by adding a very low concentration of heparin in the as-synthesized colloidal solutions. SERS-tag colloids are prepared by mixing chitosan with HAuCl4 and 4-mercapto benzoic acid before being reduced with ascorbic acid under appropriate pH conditions. The formed AuNFs-4-MBA@chitosan nanoparticles were positively charged with high stability and well-dispersed in aqueous media. Based on understanding each reaction component's role in the preparation of the SERS tag colloid, we aim to simplify the controlled synthesis and Raman probe conjugation process. The average size of AuNFs is below 90 nm, fine-tuned in shape and effectively conjugated to the Raman reporter molecules 4-MBA. These as-prepared SERS tag-based AuNFs have good biocompatibility and are virtually non-toxic, as studied with fibroblast and MCF-7 cells. Through these SERS-tag colloids, the trace detection of heparin is improved, with a wide detection window (0.01 to 100 ppm), high reproducibility (RSD value of 3.56%), limit of detection (LOD) of 0.054 ppm, and limit of quantification (LOQ) of 0.17 ppm. Comparison experiments show that the SERS-tag colloids possess good selectivity over other ions, and organic and amino acid substances. The results provide the capability and the potential for application under complex biological conditions and future biosensing based on SERS signal amplification.