Binary Thiol-Capped Gold Nanoparticle Monolayer Films for Quantitative Surface-Enhanced Raman Scattering Analysis

Surface-enhanced Raman scattering (SERS) can provide fingerprint information of analyte molecules with unparalleled sensitivity. However, quantitative analysis using SERS has remained one of the major challenges owing to the difficulty of obtaining reproducible SERS substrates with high-density hotspots. Here, we report the rational design and fabrication of a binary thiol-capped gold nanoparticle (AuNP) monolayer film (MLF) as a substrate for highly sensitive and quantitative SERS analysis. The two thiol ligands chemically bonded to the AuNPs play different roles: dodecanethiol with a long alkyl chain controls the interparticle gaps and electromagnetic coupling among AuNPs and 4-mercaptopyridine works as a Raman internal standard (IS). The binary thiol-capped AuNPs can self-assemble into an ordered MLF with high-density hotspots and uniformly distributed IS. The as-prepared MLF has been demonstrated as a reliable SERS substrate for quantitative detection of fungicide malachite green in aqueous solution, with a high enhancement factor (up to 3.3 × 10⁷) and a low detection limit (100 pM). Moreover, the MLF SERS substrate is flexible and transparent, which has enabled in situ detection of trace fungicide residues in a shrimp tissue.