{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,5]],"date-time":"2026-09-05T05:17:36Z","timestamp":1788585456120,"version":"build-2803163510"},"reference-count":30,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T00:00:00Z","timestamp":1788393600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100008736","name":"Fondo de Fomento al Desarrollo Cient\u00edfico y Tecnol\u00f3gico","doi-asserted-by":"crossref","award":["IT24I0045"],"award-info":[{"award-number":["IT24I0045"]}],"id":[{"id":"10.13039\/501100008736","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biosensors"],"abstract":"<jats:p>Background: Rapid screening of acetylcholinesterase (AChE) inhibition is essential for monitoring exposure to organophosphate compounds, particularly in field settings where access to laboratory infrastructure is limited. This study aimed to develop a portable paper-based colorimetric biosensor for the semi-quantitative detection of AChE activity in blood samples. Methods: The biosensor was based on a pH-dependent color change adapted from the modified Edson method. The platform was first optimized using experimental models and then evaluated in human capillary blood samples collected under real field conditions. Reference serum cholinesterase activity was determined by a certified clinical laboratory and used for comparison with the colorimetric response of the biosensor. RGB (red, green, and blue) image analysis was performed in a subset of samples to digitally characterize the chromatic response of the device. Results: Samples with preserved AChE activity showed a visible color shift associated with substrate hydrolysis, whereas samples with reduced or inhibited activity maintained darker blue-dominant tones. RGB analysis of human samples revealed significant associations between AChE activity and the R and G channels, supporting the ability of the platform to distinguish between chromatic patterns associated with different ranges of enzymatic activity. Conclusions: The proposed platform demonstrated the feasibility of translating a pH-based AChE assay into a portable paper-based format for semi-quantitative visual and RGB-assisted screening. Its evaluation in human samples demonstrated the feasibility of its use under field conditions as a proof-of-concept. Further studies are needed to optimize its analytical performance and validate its application in larger and more diverse populations.<\/jats:p>","DOI":"10.3390\/bios16090490","type":"journal-article","created":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T10:10:33Z","timestamp":1788430233000},"page":"490","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Portable Paper-Based Colorimetric Biosensor for Rapid Screening of Organophosphate Exposure via Acetylcholinesterase Activity and RGB Image Analysis"],"prefix":"10.3390","volume":"16","author":[{"given":"Carlos E.","family":"Zambra","sequence":"first","affiliation":[{"name":"Department of Industrial Technologies, Faculty of Engineering, University of Talca, Curic\u00f3 3640000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5257-4511","authenticated-orcid":false,"given":"Jorge","family":"Morales-Ferreiro","sequence":"additional","affiliation":[{"name":"School of Science, Engineering and Technology, School of Engineering, Universidad Mayor, Santiago 7500994, Chile"},{"name":"Center for Applied Nanotechnology (CNAP), School of Science, Engineering and Technology, Universidad Mayor, Santiago 7500994, Chile"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]},{"given":"Francisca","family":"Herrera Vielma","sequence":"additional","affiliation":[{"name":"Pharmacology Laboratory, Biomedical Sciences Department, Faculty of Health Sciences, University of Talca, Talca 3465548, Chile"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2930-991X","authenticated-orcid":false,"given":"Jessica","family":"Z\u00fa\u00f1iga-Hern\u00e1ndez","sequence":"additional","affiliation":[{"name":"Pharmacology Laboratory, Biomedical Sciences Department, Faculty of Health Sciences, University of Talca, Talca 3465548, Chile"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"40802","DOI":"10.1039\/D5RA05552K","article-title":"Organophosphate pesticides: A review on classification, synthesis, toxicity, remediation and analysis","volume":"15","author":"Saad","year":"2025","journal-title":"RSC Adv."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"96002","DOI":"10.1289\/EHP9934","article-title":"Health effects of pesticide exposure in Latin American and the Caribbean populations: A scoping review","volume":"130","author":"Hyland","year":"2022","journal-title":"Environ. 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