Machine learning-assisted surface-enhanced raman spectroscopy for the rapid determination of the glutathione redox ratio

Abstract

Rapid and accurate detection of glutathione in its reduced (GSH) and oxidized (GSSG) forms is essential for monitoring oxidative stress in biological systems. Oxidative stress is a key indicator of various diseases, and glutathione plays a vital role in maintaining the balance between oxidative and anti-oxidative processes. Surface-enhanced Raman spectroscopy (SERS) offers a highly sensitive and selective analytical approach for detecting biomolecules. However, complex biological matrices and molecules with similar chemical structure (such as GSH and GSSG) often result in overlapping vibrational signatures, making it challenging to quantify the GSH : GSSG ratio. To address this challenge, we integrated machine learning (ML) algorithms with SERS to accurately quantify the GSH : GSSG ratio in aqueous solutions. Three machine learning algorithms – support vector regression (SVR), extreme gradient boosting (XGBoost), and multilayer perceptron (MLP) were trained and evaluated using preprocessed SERS spectra of mixtures of various GSH : GSSG ratios. Among these models, MLP exhibits the highest accuracy and robustness with correlation coefficient for the test set (Q²) value of 0.966. This study highlights a practical protocol for leveraging machine learning and SERS to achieve rapid, and accurate determination of glutathione redox ratios.