Surface-enhanced Raman spectroscopy (SERS) is a powerful tool with high potential for multiplexed detection of dilute analytes. However, quantitative SERS of kinetic assays can be difficult due to the variation in enhancement factors caused by changing reaction conditions. We report a method for quantitative SERS kinetic analysis using colloidal Ag-Au core-shell nanocubes (Ag@AuNCs) as the SERS substrate. This substrate is mass producible, possesses large SERS enhancement, and is resistant to degradation in most environments. The SERS enhancement of the Ag@AuNCs was evaluated both experimentally and computationally. Quantitation was achieved by covalently attaching a non-reactive internal standard (IS) to substrate surfaces and normalizing SERS spectra to the IS signal. We demonstrated that IS normalization corrects for temporal variations in enhancement factor and particle concentration. Quantitation was demonstrated by monitoring the base-catalyzed aldol condensation of surface-bound 4-(methylthio)benzaldehyde with free acetone. The kinetic model of this reaction was fitted to IS normalized SERS data, resulting in kinetic parameters that agreed well with published values. This SERS platform is a robust and sensitive method for quantitative analysis of kinetic assays, with potential applications in many fields.
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