Simultaneous quantitative analysis of multiple metabolites using label-free surface-enhanced Raman spectroscopy and explainable deep learning

Abstract

Metabolites serve as vital biomarkers, reflecting physiological and pathological states and offering insights into disease progression and early detection. This study introduces an advanced analytical technique integrating label-free Surface-Enhanced Raman Spectroscopy (SERS) with deep learning, and leverages SHAP (SHapley Additive exPlanations) to provide a visual interpretative analysis of the predictive rationale of the deep learning model, facilitating simultaneous detection and quantitative analysis of multiple metabolites. Monolayer silver nanoparticle SERS substrates were fabricated via a triple-phase interfacial self-assembly method, which captured complex spectral information of target metabolites in mixed solutions. A custom-built deep neural network model with multi-channel feature extraction was employed to predict the concentrations of uric acid (R² = 0.976), xanthine (R² = 0.971), hypoxanthine (R² = 0.977), and creatinine (R² = 0.940). The method’s scalability was validated as the performance remained consistent with an increasing number of simultaneous targets. This approach offers a sensitive, cost-effective, and rapid alternative for metabolite analysis, with significant implications for clinical diagnostics and personalized medicine.