{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T14:17:11Z","timestamp":1785421031307,"version":"3.56.0"},"reference-count":31,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T00:00:00Z","timestamp":1782950400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001807","name":"S\u00e3o Paulo Research Foundation","doi-asserted-by":"crossref","award":["2020\/07451-1"],"award-info":[{"award-number":["2020\/07451-1"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"crossref"}]},{"award":["2020\/07451-1"],"award-info":[{"award-number":["2020\/07451-1"]}],"id":[{"id":"https:\/\/ror.org\/02ddkpn78","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxics"],"abstract":"<jats:p>The increasing prohibition of animal testing for cosmetic products has driven the development of alternative approaches to ensure consumer safety. Skin sensitization is one of the most critical toxicological endpoints to evaluate, requiring rigorous assessment to ensure the safety of cosmetic ingredients. This study proposes an Integrated Testing Strategy (ITS) that combines in chemico (Direct Peptide Reactivity Assay\u2014DPRA) and in silico approaches (a six-platform computational panel) to evaluate the sensitization potential of substances. Initially, the in chemico methodology was validated through a partial proficiency demonstration to ensure experimental reliability. Subsequently, this ITS was applied to the Baccharis trimera extract and its major marker, 3-Caffeoylquinic acid (chlorogenic acid\/3-CQA). Our results demonstrate that the DPRA alone is insufficient to classify the sensitization potential of complex mixtures, as recommended by OECD guidelines. The integration of in silico data proved essential to interpret the reactivity of the botanical matrix, revealing that the sensitization potential observed in the extract does not stem solely from 3-CQA, but likely results from the synergistic contribution of more lipophilic caffeoylquinic acid isomers. This approach demonstrates that integrating experimental and computational methods is fundamental for a robust safety assessment, offering an efficient, animal-free strategy for the early screening of cosmetic ingredients and for refining the interpretation of toxicological data in complex chemical environments.<\/jats:p>","DOI":"10.3390\/toxics14070585","type":"journal-article","created":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T14:51:25Z","timestamp":1783003885000},"page":"585","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Animal-Free Skin Sensitization Testing: In Chemico and In Silico Integrated Approach"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6984-375X","authenticated-orcid":false,"given":"Gabriella","family":"Lisboa dos Santos","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Sciences, Federal University of S\u00e3o Paulo, Diadema Campus, Diadema 09913-030, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1786-4756","authenticated-orcid":false,"given":"Gabriela de Oliveira Prado","family":"Corr\u00eaa","sequence":"additional","affiliation":[{"name":"Pre-Clinical & Product Safety-Grupo Botic\u00e1rio, R. Alfredo Pinto, 2258-Parque da Fonte, S\u00e3o Jos\u00e9 dos Pinhais 83050-320, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Franciane de Oliveira","family":"Cortez","sequence":"additional","affiliation":[{"name":"Pre-Clinical & Product Safety-Grupo Botic\u00e1rio, R. Alfredo Pinto, 2258-Parque da Fonte, S\u00e3o Jos\u00e9 dos Pinhais 83050-320, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tugst\u00eanio","family":"Souza","sequence":"additional","affiliation":[{"name":"Pre-Clinical & Product Safety-Grupo Botic\u00e1rio, R. Alfredo Pinto, 2258-Parque da Fonte, S\u00e3o Jos\u00e9 dos Pinhais 83050-320, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-4974-8832","authenticated-orcid":false,"given":"Bruna","family":"Bosquetti","sequence":"additional","affiliation":[{"name":"Pre-Clinical & Product Safety-Grupo Botic\u00e1rio, R. Alfredo Pinto, 2258-Parque da Fonte, S\u00e3o Jos\u00e9 dos Pinhais 83050-320, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jo\u00e3o Antonio Dassie","family":"Felippi","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, Federal University of S\u00e3o Paulo, Diadema Campus, Diadema 09913-030, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1170-9678","authenticated-orcid":false,"given":"Gabriela Trindade de Souza","family":"e Silva","sequence":"additional","affiliation":[{"name":"Faculdade de Ci\u00eancias Farmac\u00eauticas, State University of Campinas, Rua C\u00e2ndido Portinari, 200, Cidade Universit\u00e1ria Zeferino Vaz, Campinas 13083-871, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7507-8393","authenticated-orcid":false,"given":"Pamela Ferreira","family":"do Nascimento","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, Federal University of S\u00e3o Paulo, Diadema Campus, Diadema 09913-030, Brazil"},{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Medicina Translacional, Departamento de Medicina, Escola Paulista de Medicina, Universidade Federal de S\u00e3o Paulo, S\u00e3o Paulo 04021-001, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6372-3677","authenticated-orcid":false,"given":"Marcio Adriano","family":"Andr\u00e9o","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, Federal University of S\u00e3o Paulo, Diadema Campus, Diadema 09913-030, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4080-9862","authenticated-orcid":false,"given":"Marcelo Dutra","family":"Duque","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, Federal University of S\u00e3o Paulo, Diadema Campus, Diadema 09913-030, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carolina Motter","family":"Catarino","sequence":"additional","affiliation":[{"name":"Pre-Clinical & Product Safety-Grupo Botic\u00e1rio, R. Alfredo Pinto, 2258-Parque da Fonte, S\u00e3o Jos\u00e9 dos Pinhais 83050-320, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrezza di Pietro Micali","family":"Canavez","sequence":"additional","affiliation":[{"name":"Pre-Clinical & Product Safety-Grupo Botic\u00e1rio, R. Alfredo Pinto, 2258-Parque da Fonte, S\u00e3o Jos\u00e9 dos Pinhais 83050-320, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1353-9825","authenticated-orcid":false,"given":"Patricia Santos","family":"Lopes","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, Federal University of S\u00e3o Paulo, Diadema Campus, Diadema 09913-030, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1111\/cod.13294","article-title":"Evaluation of in Vitro Testing Strategies for Hazard Assessment of the Skin Sensitization Potential of \u201cReal-Life\u201d Mixtures: The Case of Henna-Based Hair-Colouring Products Containing p-Phenylenediamine","volume":"81","author":"Veloso","year":"2019","journal-title":"Contact Dermat."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1152","DOI":"10.1111\/j.1398-9995.2011.02652.x","article-title":"Mechanisms of Chemical-Induced Innate Immunity in Allergic Contact Dermatitis","volume":"66","author":"Martin","year":"2011","journal-title":"Allergy"},{"key":"ref_3","unstructured":"OECD (2025). 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Pharmacol."}],"container-title":["Toxics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2305-6304\/14\/7\/585\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T10:02:01Z","timestamp":1783418521000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2305-6304\/14\/7\/585"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,2]]},"references-count":31,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["toxics14070585"],"URL":"https:\/\/doi.org\/10.3390\/toxics14070585","relation":{},"ISSN":["2305-6304"],"issn-type":[{"value":"2305-6304","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,2]]}}}