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In this study, GC\u00d7GC-TOFMS was combined with an image-based chromatographic fingerprinting strategy to characterize the fecal volatilome of subjects affected by non-celiac gluten\/wheat sensitivity (NCGWS) under dietary intervention. Fifty participants underwent a dietary protocol consisting of a gluten-free diet followed by gluten reintroduction while receiving probiotics or placebo. Headspace solid-phase microextraction (HS-SPME) conditions were specifically optimized to maximize the extraction coverage of volatiles, while a combined untargeted\u2013targeted fingerprinting approach (\n                    <jats:italic>UT fingerprinting<\/jats:italic>\n                    ) enabled the mapping of approximately 1000 volatile features in fecal samples. Chemometric analyses, including principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA), highlighted informative differences in VOC profiles induced by gluten reintroduction and probiotic administration. Around 270 VOCs were annotated, expanding the existing knowledge on the human fecal volatilome by 20%, predominantly comprising microbial fermentation-derived metabolites such as short-chain fatty acid esters, aldehydes, alcohols, terpenes, and aromatic hydrocarbons. PLS-DA models discriminated probiotic-treated individuals from placebo recipients with 89\u201390% accuracy, confirming the impact of probiotic supplementation on gut metabolic signatures. The integration of GC\u00d7GC-TOFMS and image-based chromatographic fingerprinting effectively captured subtle metabolic variations, demonstrating its suitability in biomarker discovery for dietary interventions and personalized nutrition studies.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Graphical abstract<\/jats:bold>\n                  <\/jats:p>","DOI":"10.1007\/s00216-025-06280-6","type":"journal-article","created":{"date-parts":[[2026,1,8]],"date-time":"2026-01-08T09:12:58Z","timestamp":1767863578000},"page":"3037-3058","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Advancing fecal volatilome profiling by comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry and image pattern recognition"],"prefix":"10.1007","volume":"418","author":[{"given":"Fulvia","family":"Trapani","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrea","family":"Caratti","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Erica","family":"Liberto","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luca","family":"Cocolin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ilaria","family":"Goitre","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Valentina","family":"Ponzo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Simona","family":"Bo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3201-0775","authenticated-orcid":false,"given":"Chiara","family":"Cordero","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ilario","family":"Ferrocino","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2026,1,8]]},"reference":[{"key":"6280_CR1","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1016\/0021-9673(95)00249-M","volume":"703","author":"JC Giddings","year":"1995","unstructured":"Giddings JC. 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