Hybrid Machine Learning-Assisted Raman Spectroscopy with Generative Feature Augmentation for Pharmaceutical Identification

Raman spectroscopy enables label-free molecular characterization across materials science, analytical chemistry, biomedicine, and industrial process monitoring. However, machine learning for high-dimensional spectroscopy remains constrained by limited labelled data and a mismatch between the physical organization of spectra and feature-agnostic models. Channel coverage alone does not ensure that related bands share a common inference context. Here we present RamanPFN, a general-purpose spectral foundation framework that enables unified in-context inference through physics-guided spectral learning. It captures full-spectrum compositional covariation via Global Compositional Unmixing (GCU), which decomposes distributed, multi-band mixture signatures into shared non-negative latent bases. Simultaneously, it resolves local vibrational structure through Local Vibrational Subspace Encoding (LVSE), which preserves fine-grained peak morphology, intensity fluctuations, and peak shifts within contiguous spectral neighborhoods. Extensive evaluation across 74 diverse public Raman datasets covered 129 regression targets and was further extended to 21 classification tasks. RamanPFN achieved state-of-the-art performance across all reported aggregate metrics against 28 independently reproduced methods spanning chemometrics, spectral neural networks, deep tabular learners and tabular foundation models. RamanPFN establishes a physics-guided paradigm for scientific spectroscopy, enabling data-efficient predictive learning across diverse chemical systems.

Access to sufficiently large biomedical datasets remains a major obstacle for machine learning in Raman spectroscopy-based diagnostics. In particular, for glioma analysis, datasets are typically small and heterogeneous, affected by acquisition-specific variability. This work investigates the utility of deep generative augmentation in such a small-cohort setting. We analyze glioma biopsy spectra acquired from 58 tumor samples and consider both binary IDH-status classification and 6-class methylation subtype classification problems. To address the limited size and imbalance of the dataset, we develop a conditional variational autoencoder ($β$-CVAE) capable of generating class-conditioned synthetic Raman spectra. The generated data are evaluated in Train-on-Synthetic, Test-on-Real (TS/TR) and Train-on-Synthetic+Real, Test-on-Real (TSR/TR) settings under a strict patient-isolated cross-validation protocol. Models trained exclusively on synthetic data underperform models trained on real spectra, indicating a substantial domain gap between synthetic and real distributions. However, augmenting the real training data with synthetic spectra consistently improves classification performance across multiple models. These findings indicate that, even with a limited number of independent patient samples, generative models can capture sufficient structure to provide useful regularization for downstream classifiers. We also investigate a reconstruction-based inference strategy, termed Classification by Reconstruction (CbR), in which class prediction is based on reconstruction error under different class conditions. Overall, the results support the use of deep generative augmentation as a practical strategy for improving machine learning robustness in Raman spectroscopy applications characterized by limited biomedical datasets.

Antimicrobial resistance is expected to claim 10 million lives per year by 2050, and resource-limited regions are most affected. Raman spectroscopy is a novel pathogen diagnostic approach promising rapid and portable antibiotic resistance testing within a few hours, compared to days when using gold standard methods. However, current algorithms for Raman spectra analysis 1) are unable to generalize well on limited datasets across diverse patient populations and 2) require increased complexity due to the necessity of non-trivial pre-processing steps, such as feature extraction, which are essential to mitigate the low-quality nature of Raman spectral data. In this work, we address these limitations using Sharpness-Aware Minimization (SAM) to enhance model generalization across a diverse array of hyperparameters in clinical bacterial isolate classification tasks. We demonstrate that SAM achieves accuracy improvements of up to 10.5% on a single split, and an increase in average accuracy of 2.7% across all splits in spectral classification tasks over the traditional optimizer, Adam. These results display the capability of SAM to advance the clinical application of AI-powered Raman spectroscopy tools.