Hamadouche D. auteur aut IRD Diarra Adama Zan auteur aut IRD Fohrer F. auteur aut IRD Bérenger J. M. auteur aut IRD Benakhla A. auteur aut IRD Almeras L. auteur aut IRD Parola P. auteur aut IRD text journalArticle eng text/pdf reformatted digital access When heritage properties and contents are threatened by various harmful insects, especially coleopterans, rapid and reliable pest identification is required for effective management. While traditional methods have limitations, MALDI-TOF MS has recently emerged as a groundbreaking identification tool. In this study, our aim was to assess the effectiveness of MALDI-TOF MS in identifying cultural heritage insect pests belonging to the Coleoptera order. To do so, adult, larva, and exuviae from eight species of Coleoptera (Lasioderma serricorne (n = 92), Attagenus smirnovi (n = 87), Trogoderma versicolor (n = 97), Reesa vespulae (n = 74), Anthrenus verbasci (n = 68), Anthrenus pimpinellae (n = 34), Anthrenus flavipes (n = 30), and Thylodrias contractus (n = 19), were subjected to MALDI-TOF MS after optimisation of the sample preparation parameters. This optimisation was based on three key parameters influencing spectrum quality: the choice of body part, the buffer volume for protein extraction, and the method and duration of sample homogenisation. Reproducible intra-species MS spectra for each developmental stage were obtained. The species specificity of MS spectra for each developmental stage supported the use of MALDI- TOF MS for specimen classification. Spectra from 68 samples, including four specimens per species and stage, were added to our homemade reference arthropod MS database (DB). Among spectra classified as being consistent, 97.10% (335/345) of the samples were correctly classified with a relevant identification score. The morphological identification of all adult specimens in the reference MS spectra DB was confirmed by molecular biology, and the sequences were deposited in GenBank. This study is the first demonstration of the effective application of MALDI-TOF MS as a reliable tool for identifying various developmental stages of Coleoptera pests which pose a threat to the cultural heritage of France. specialized Coleopteran Heritage MALDI-TOF MS Entomology Pest FRANCE 080 020 084 199 106033 [11 ] 2025 0964-8305 https://www.documentation.ird.fr/hor/fdi:010092868 10.1016/j.ibiod.2025.106033 0964-8305 [F B010092868] https://www.documentation.ird.fr/hor/fdi:010092868 https://horizon.documentation.ird.fr/exl-doc/pleins_textes/2025-04/010092868.pdf IRD - Base Horizon / Pleins textes 2025-04-02 2025-11-04 fdi:010092868 fre