Integration of mass spectrometry and molecular biotechnology to study bioaerosols

Conventional culture-based and microscopic approaches yield limited information about the diversity, content, and real-time behaviour of biological aerosols. In recent years, mass spectrometry (MS) and molecular biotechnology have evolved as powerful and complementary analytical methods for detecting, identifying, and characterising air biological particles. This study critically reviews recent improvements in MS-based techniques for analysing bioaerosol chemical markers, proteins, metabolites, and toxins, including MALDI-TOF MS, GC-MS, LC-MS/MS, and real-time aerosol mass spectrometry. In parallel, contemporary advances in molecular biotechnology, including as PCR-based assays, metagenomics, and MS-driven proteomics and metabolomics, are described, with a focus on atmospheric applications. Special emphasis is placed on integrated analytical workflows that combine MS with molecular techniques to improve specificity, sensitivity, and source attribution. The current issues of low biomass concentrations, sampling artefacts, data interpretation, and standardisation are discussed, and future perspectives on portable MS systems, multi-omics integration, and AI-assisted data processing are presented. This study offers a thorough analytical chemistry viewpoint on next-generation methodologies for monitoring bioaerosols and promotes the development of enhanced instruments for assessing air quality and protecting human health.