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Although several clinical methods are available for the diagnosis of candidiasis, they have limitations related to response time and laboratory complexity. In this study, spectroscopic techniques based on excitation\u2013emission matrix (EEM) fluorescence were explored in combination with higher-order chemometric methods, such as PARAFAC and Tucker3, coupled with classification algorithms, including Quadratic Discriminant Analysis (QDA), for the identification of seven Candida classes: C. albicans, C. auris, C. glabrata, C. hemeulonii, C. krusei, C. parapsilosis, and C. tropicalis. The analyses were conducted using a limited data set consisting of seven Candida species measured in ten independent replicates, resulting in 70 data points. PARAFAC-QDA and Tucker3-QDA models were developed for multiclass classification. Although evaluated on a small-scale sample size, both models reached 100% classification accuracy, sensitivity, and specificity for training and test set, highlighting the potential of multiway analysis for early exploration. The selected excitation and emission wavelengths associated with the latent factors were mainly distributed in the excitation region of 260, 270, and 280 nm, and in the emission range of 280 to 350 nm, which are strongly related to aromatic amino acids, such as tyrosine, tryptophan, and phenylalanine, reflecting the structural composition of these microorganisms. These results demonstrate the potential of the proposed approach as a proof-of-concept for the discrimination of reference Candida strains under controlled culture conditions, supporting future validation using independent clinical isolates.<\/jats:p>","DOI":"10.1021\/acsomega.6c03805","type":"journal-article","created":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T17:02:47Z","timestamp":1786726967000},"page":"51180-51190","source":"Crossref","is-referenced-by-count":0,"title":["Second-Order Chemometric Classification of\n                    <i>Candida<\/i>\n                    Species Using Excitation\u2013Emission Matrix\nFluorescence Spectroscopy: A Proof-of-Concept Study"],"prefix":"10.1021","volume":"11","author":[{"given":"Lavi\u0301nia H. S.","family":"Pereira","sequence":"first","affiliation":[{"name":"Federal University of Rio Grande do Norte , , , ,","place":["Natal, RN, Brazil, 5072-970"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria R. C.","family":"Ina\u0301cio","sequence":"additional","affiliation":[{"name":"Federal University of Rio Grande do Norte , , , ,","place":["Natal, RN, Brazil, 5072-970"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francimara C. S.","family":"Silvestre","sequence":"additional","affiliation":[{"name":"Federal University of Rio Grande do Norte , , , ,","place":["Natal, RN, Brazil, 5072-970"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rafael W.","family":"Bastos","sequence":"additional","affiliation":[{"name":"Federal University of Rio Grande do Norte , , , ,","place":["Natal, RN, Brazil, 5072-970"]},{"name":"National Institute of Science and Technology in Human Pathogenic Fungi , , ,","place":["Ribeira\u0303o Preto, Brazil, 14040-900"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3827-3800","authenticated-orcid":true,"given":"Ka\u0301ssio M. G.","family":"Lima","sequence":"additional","affiliation":[{"name":"Federal University of Rio Grande do Norte , , , ,","place":["Natal, RN, Brazil, 5072-970"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,14]]},"reference":[{"issue":"165","key":"2026090307191305400_cit1","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.3004404","article-title":"Hidden killers:\nhuman fungal infections","volume":"4","author":"Brown","year":"2012","journal-title":"Sci. Transl Med."},{"issue":"3","key":"2026090307191305400_cit2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00449-20","article-title":"Threats posed by the\nfungal kingdom to humans, wildlife, and agriculture","volume":"11","author":"Fisher","year":"2020","journal-title":"mBio"},{"issue":"1","key":"2026090307191305400_cit3","doi-asserted-by":"publisher","DOI":"10.1038\/nrdp.2018.26","article-title":"Invasive candidiasis","volume":"4","author":"Pappas","year":"2018","journal-title":"Nat. Rev.\nDis Primers."},{"key":"2026090307191305400_cit4","doi-asserted-by":"publisher","first-page":"1198","DOI":"10.1056\/NEJMoa1306801","article-title":"Multistate point-prevalence\nsurvey of health careassociated infections","volume":"370","author":"Magill","year":"2014","journal-title":"N. Engl. J. Med."},{"issue":"9","key":"2026090307191305400_cit5","doi-asserted-by":"publisher","first-page":"1172","DOI":"10.1086\/378745","article-title":"Attributable mortality\nof nosocomial candidemia, revisited","volume":"37","author":"Gudlaugsson","year":"2003","journal-title":"Clin. Infect.\nDis."},{"key":"2026090307191305400_cit6","doi-asserted-by":"publisher","first-page":"174","DOI":"10.3201\/eid0702.010203","article-title":"The impact of hospital-acquired bloodstream infections","volume":"7","author":"Wenzel","year":"2001","journal-title":"Emerg. Infect. Dis."},{"issue":"9","key":"2026090307191305400_cit7","doi-asserted-by":"publisher","first-page":"3640","DOI":"10.1128\/AAC.49.9.3640-3645.2005","article-title":"Delaying the empiric treatment of\nCandida bloodstream infection until positive blood results are obtained:\na potential risk factor for hospital mortality","volume":"49","author":"Morrell","year":"2005","journal-title":"Antimicrob. Agents Chemother."},{"issue":"1","key":"2026090307191305400_cit8","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1086\/504810","article-title":"Time to initiation of\nflucanazole therapy impacts mortality in patients with candidemia:\na multi-institutional study","volume":"43","author":"Garey","year":"2006","journal-title":"Clin. Infect. Dis."},{"issue":"1","key":"2026090307191305400_cit9","doi-asserted-by":"publisher","DOI":"10.1117\/1.JBO.22.1.016002","article-title":"Hyperspectral\nfluorescence microscopy\ndetects autofluorescent factors that can be exploited as a diagnostic\nmethod for Candida species differentiation","volume":"22","author":"Graus","year":"2017","journal-title":"J. Biomed. 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Biomol.\nSpectrosc."},{"key":"2026090307191305400_cit27","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1016\/j.foodchem.2018.04.112","article-title":"Determination\nof toxic elements in\nyerba mate by ICP-MS after diluted acid digestion under O2 pressure","volume":"263","author":"Pardinho","year":"2018","journal-title":"Food Chem."},{"issue":"1\u20133","key":"2026090307191305400_cit28","doi-asserted-by":"publisher","first-page":"71","DOI":"10.1016\/S0003-2670(97)00065-2","article-title":"Genetic\nalgorithms as a method for variable selection in multiple linear regression\nand partial least squares regression, with applications to pyrolysis\nmass spectrometry","volume":"348","author":"Broadhurst","year":"1997","journal-title":"Anal. Chim. Acta"},{"issue":"3","key":"2026090307191305400_cit29","doi-asserted-by":"publisher","first-page":"279","DOI":"10.1007\/BF02289464","article-title":"Some Mathematical\nNotes on Three-mode Factor Analysis","volume":"31","author":"Tucker","year":"1966","journal-title":"Psychometrika"},{"issue":"1","key":"2026090307191305400_cit30","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/0169-7439(93)E0086-J","article-title":"N-way principal\ncomponent analysis theory, algorithms and applications","volume":"25","author":"Henrion","year":"1994","journal-title":"Chemom. Intell. Lab. Syst."},{"key":"2026090307191305400_cit31","doi-asserted-by":"crossref","DOI":"10.1002\/0470863242","volume-title":"Chemometrics: Data\nAnalysis for the Laboratory and Chemical Plant","author":"Brereton","year":"2003"},{"issue":"1\u20132","key":"2026090307191305400_cit32","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1016\/S0169-7439(01)00143-5","article-title":"A robust version\nof the Tucker3 model","volume":"59","author":"Pravdova","year":"2001","journal-title":"Chemom. Intell. Lab. Syst."},{"issue":"5","key":"2026090307191305400_cit33","doi-asserted-by":"publisher","first-page":"910","DOI":"10.1590\/S0100-40422005000500032","article-title":"PARAFAC: uma ferramenta\nquimiome\u0301trica\npara tratamento de dados multidimensionais. Aplicac\u0327o\u0303es\nna determinac\u0327a\u0303o direta de fa\u0301rmacos em plasma\nhumano por espectrofluorimetria","volume":"28","author":"Sena","year":"2005","journal-title":"Qui\u0301mica\nNova"},{"key":"2026090307191305400_cit34","volume-title":"Kronecker Products and\nMatrix Calculus: With Applications","author":"Graham","year":"1981"},{"key":"2026090307191305400_cit35","doi-asserted-by":"publisher","first-page":"46","DOI":"10.1016\/j.chemolab.2019.03.007","article-title":"TTWD-DA:\nA MATLAB toolbox for discriminant\nanalysis based on trilinear three-way data","volume":"188","author":"Morais","year":"2019","journal-title":"Chemom. Intell. Lab. Syst."},{"key":"2026090307191305400_cit36","doi-asserted-by":"publisher","first-page":"151","DOI":"10.1016\/j.trac.2018.07.022","article-title":"Modeling\nsecond-order data for classification issues:\nData characteristics, algorithms, processing procedures and applications","volume":"107","author":"Azcarate","year":"2018","journal-title":"TrAC, Trends Anal. Chem."},{"issue":"1","key":"2026090307191305400_cit37","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.chemolab.2008.07.010","article-title":"Comparison of performance\nof five common classifiers\nrepresented as boundary methods: Euclidean Distance to Centroids,\nLinear Discriminant Analysis, Quadratic Discriminant Analysis, Learning\nVector Quantization and Support Vector Machines, as dependent on data\nstructure","volume":"95","author":"Dixon","year":"2009","journal-title":"Chemom. Intell. Lab. Syst."},{"key":"2026090307191305400_cit38","doi-asserted-by":"publisher","DOI":"10.1016\/j.actatropica.2022.106779","article-title":"Detection of terbufos in cases of\nintoxication by means of entomotoxicological analysis using ATR-FTIR\nspectroscopy combined with chemometrics","volume":"238","author":"Silva","year":"2023","journal-title":"Acta\nTrop."},{"issue":"1","key":"2026090307191305400_cit39","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.chemolab.2008.07.010","article-title":"Comparison of performance\nof five common classifiers\nrepresented as boundary methods: Euclidean Distance to Centroids,\nLinear Discriminant Analysis, Quadratic Discriminant Analysis, Learning\nVector Quantization and Support vector Machines, as dependent on data\nstructure","volume":"95","author":"Dixon","year":"2009","journal-title":"Chemom. Intell. Lab. 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Rep."},{"issue":"10","key":"2026090307191305400_cit42","doi-asserted-by":"publisher","first-page":"869","DOI":"10.1080\/05704928.2023.2165091","article-title":"A review of biospectroscopy coupled\nwith chemometrics for Alzheimer\u2019s disease diagnosis","volume":"58","author":"Santos","year":"2023","journal-title":"Appl. Spectrosc. Rev."},{"key":"2026090307191305400_cit43","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.chemolab.2017.09.001","article-title":"Comparing unfolded and two-dimensional\ndiscriminant\nanalysis and support vector machines for classification of EEM data","volume":"170","author":"Morais","year":"2017","journal-title":"Chemom. Intell. Lab. Syst."},{"key":"2026090307191305400_cit44","doi-asserted-by":"publisher","first-page":"74","DOI":"10.1016\/j.trac.2012.02.005","article-title":"Multivariate class\nmodeling for the verification of\nfood-authenticity claims","volume":"35","author":"Oliveri","year":"2012","journal-title":"TrAC, Trends Anal.\nChem."},{"issue":"10","key":"2026090307191305400_cit45","doi-asserted-by":"publisher","DOI":"10.3390\/photonics9100692","article-title":"Excitation-Dependent\nFluorescence Helps to Indicate Fungal Contamination of Aquatic Environments\nand to Differentiate Filamentous Fungi","volume":"9","author":"Fedoseeva","year":"2022","journal-title":"Photonics"},{"key":"2026090307191305400_cit46","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2023.1131122","article-title":"A whole cell fluorescence\nquenching-based approach for the investigation of polyethyleneimine\nfunctionalized silver nanoparticles interaction with Candida albicans","volume":"14","author":"Tiwari","year":"2023","journal-title":"Front. Microbiol."},{"key":"2026090307191305400_cit47","doi-asserted-by":"publisher","DOI":"10.21577\/0103-5053.20260027","article-title":"Excitation-Emission Matrix Fluorescence\nof Blood Plasma Combined with Multi-Way Chemometric Analysis for HIV\nDetection in Pregnant Women","author":"Silva","year":"2026","journal-title":"J. Braz. Chem.\nSoc."},{"issue":"5","key":"2026090307191305400_cit48","doi-asserted-by":"publisher","first-page":"10271","DOI":"10.3390\/s150510271","article-title":"Fluorescence Spectroscopy\nand Chemometric Modeling\nfor Bioprocess Monitoring","volume":"15","author":"Faassen","year":"2015","journal-title":"Sensors"},{"issue":"1","key":"2026090307191305400_cit49","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-020-70811-7","article-title":"Molecular fluorescence\nspectroscopy with Multi-way analysis techniques detects spectral variations\ndistinguishing uninfected serum versus dengue or chikungunya viral\ninfected samples","volume":"10","author":"Santos","year":"2020","journal-title":"Sci. Rep."},{"key":"2026090307191305400_cit50","doi-asserted-by":"publisher","first-page":"7","DOI":"10.1016\/B978-0-444-63527-3.00001-1","article-title":"Fundamentals of PARAFAC","volume-title":"Data Handling\nin Science and Technology","author":"Marti\u0301","year":"2015"},{"issue":"1","key":"2026090307191305400_cit51","doi-asserted-by":"publisher","DOI":"10.1117\/1.JBO.22.1.016002","article-title":"Hyperspectral Fluorescence\nMicroscopy\nDetects Autofluorescent Factors that can be Exploited as a Diagnostic\nMethod for Candida species differentiation","volume":"22","author":"Graus","year":"2017","journal-title":"J. Biomed. Opt."},{"issue":"5","key":"2026090307191305400_cit52","doi-asserted-by":"publisher","first-page":"910","DOI":"10.1590\/S0100-40422005000500032","article-title":"PARAFAC:\nUma ferramenta quimiome\u0301trica\npara tratamento de dados multidimensionais. Aplicac\u0327o\u0303es\nna determinac\u0327a\u0303o direta de fa\u0301rmacos em plasma\nhumano por espectrofluorimetria","volume":"28","author":"Sena","year":"2005","journal-title":"Qui\u0301m.\nNova"},{"key":"2026090307191305400_cit53","doi-asserted-by":"publisher","DOI":"10.1016\/j.jfca.2024.106212","article-title":"Evolving trends\nin fluorescence spectroscopy techniques for food\nquality and safety: A review","volume":"131","author":"Gu","year":"2024","journal-title":"J. Food Compos.\nAnal."}],"container-title":["ACS Omega"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acsomega.6c03805","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/acsodf\/article-pdf\/11\/34\/51180\/66554754\/acsomega.6c03805.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/acsodf\/article-pdf\/11\/34\/51180\/66554754\/acsomega.6c03805.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T11:19:22Z","timestamp":1788434362000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/acsodf\/article\/11\/34\/51180\/5256013\/Second-Order-Chemometric-Classification-of-Candida"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,14]]},"references-count":53,"journal-issue":{"issue":"34","published-online":{"date-parts":[[2026,8,14]]},"published-print":{"date-parts":[[2026,9,1]]}},"URL":"https:\/\/doi.org\/10.1021\/acsomega.6c03805","relation":{},"ISSN":["2470-1343"],"issn-type":[{"value":"2470-1343","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,14]]}}}