Despite decades of progress in machine learning applications for predicting molecular structures from MS/MS spectra, the development of new methods is severely hindered by the lack of standard datasets and evaluation protocols. To address this problem, we propose MassSpecGym - the first comprehensive benchmark for the discovery and identification of molecules from MS/MS data.

Tandem mass spectrometry has played a pivotal role in advancing proteomics, enabling the high-throughput analysis of protein composition in biological tissues. Despite the development of several deep learning methods for predicting amino acid sequences (peptides) responsible for generating the observed mass spectra, training data biases hinder further advancements of \emph{de novo} peptide sequencing. Firstly, prior methods struggle to identify amino acids with Post-Translational Modifications (PTMs) due to their lower frequency in training data compared to canonical amino acids, further resulting in unsatisfactory peptide sequencing performance. Secondly, various noise and missing peaks in mass spectra reduce the reliability of training data (Peptide-Spectrum Matches, PSMs). To address these challenges, we propose AdaNovo, a novel and domain knowledge-inspired framework that calculates Conditional Mutual Information (CMI) between the mass spectra and amino acids or peptides, using CMI for robust training against above biases. Extensive experiments indicate that AdaNovo outperforms previous competitors on the widely-used 9-species benchmark, meanwhile yielding 3.6\% - 9.4\% improvements in PTMs identification. The supplements contain the code.

Matrix Assisted Laser Desorption/Ionization Mass Spectrometry (MALDI-MS) is a cornerstone in biomolecular analysis, offering precise identification of pathogens through unique mass spectral signatures. Yet, its reliance on labor-intensive sample preparation and multi-shot spectral averaging restricts its use to laboratory settings, rendering it impractical for real-time environmental monitoring. These limitations are especially pronounced in emerging aerosol MALDI-MS systems, where autonomous sampling generates noisy spectra for unknown aerosol analytes, requiring single-shot detection for effective analysis. Addressing these challenges, we propose the Mass Spectral Dictionary-Guided Transformer (MS-DGFormer): a data-driven framework that redefines spectral analysis by directly processing raw, minimally prepared mass spectral data. MS-DGFormer leverages a transformer architecture, designed to capture the long-range dependencies inherent in these time-series spectra. To enhance feature extraction, we introduce a novel dictionary encoder that integrates denoised spectral information derived from Singular Value Decomposition (SVD), enabling the model to discern critical biomolecular patterns from single-shot spectra with robust performance. This innovation provides a system to achieve superior pathogen identification from aerosol samples, facilitating autonomous, real-time analysis in field conditions. By eliminating the need for extensive preprocessing, our method unlocks the potential for portable, deployable MALDI-MS platforms, revolutionizing environmental pathogen detection and rapid response to biological threats.

Retrieving molecular structures from tandem mass spectra is a crucial step in rapid compound identification. Existing retrieval methods, such as traditional mass spectral library matching, suffer from limited spectral library coverage, while recent cross-modal representation learning frameworks often encounter modality misalignment, resulting in suboptimal retrieval accuracy and generalization. To address these limitations, we propose GLMR, a Generative Language Model-based Retrieval framework that mitigates the cross-modal misalignment through a two-stage process. In the pre-retrieval stage, a contrastive learning-based model identifies top candidate molecules as contextual priors for the input mass spectrum. In the generative retrieval stage, these candidate molecules are integrated with the input mass spectrum to guide a generative model in producing refined molecular structures, which are then used to re-rank the candidates based on molecular similarity. Experiments on both MassSpecGym and the proposed MassRET-20k dataset demonstrate that GLMR significantly outperforms existing methods, achieving over 40% improvement in top-1 accuracy and exhibiting strong generalizability.

A common approach to the de novo molecular generation problem from mass spectra involves a two-stage pipeline: (1) encoding mass spectra into molecular fingerprints, followed by (2) decoding these fingerprints into molecular structures. In our work, we adopt MIST (Goldman et. al., 2023) as the encoder and MolForge (Ucak et. al., 2023) as the decoder, leveraging additional training data to enhance performance. We also threshold the probabilities of each fingerprint bit to focus on the presence of substructures. This results in a tenfold improvement over previous state-of-the-art methods, generating top-1 31% / top-10 40% of molecular structures correctly from mass spectra in MassSpecGym (Bushuiev et. al., 2024). We position this as a strong baseline for future research in de novo molecule elucidation from mass spectra.

Mass spectrometry (MS) plays a critical role in molecular identification, significantly advancing scientific discovery. However, structure elucidation from MS data remains challenging due to the scarcity of annotated spectra. While large-scale pretraining has proven effective in addressing data scarcity in other domains, applying this paradigm to mass spectrometry is hindered by the complexity and heterogeneity of raw spectral signals. To address this, we propose MS-BART, a unified modeling framework that maps mass spectra and molecular structures into a shared token vocabulary, enabling cross-modal learning through large-scale pretraining on reliably computed fingerprint-molecule datasets. Multi-task pretraining objectives further enhance MS-BART's generalization by jointly optimizing denoising and translation task. The pretrained model is subsequently transferred to experimental spectra through finetuning on fingerprint predictions generated with MIST, a pre-trained spectral inference model, thereby enhancing robustness to real-world spectral variability. While finetuning alleviates the distributional difference, MS-BART still suffers molecular hallucination and requires further alignment. We therefore introduce a chemical feedback mechanism that guides the model toward generating molecules closer to the reference structure. Extensive evaluations demonstrate that MS-BART achieves SOTA performance across 5/12 key metrics on MassSpecGym and NPLIB1 and is faster by one order of magnitude than competing diffusion-based methods, while comprehensive ablation studies systematically validate the model's effectiveness and robustness.