Academic Journal
Quantitative TLC-based assay for the detection of urease inhibitors in complex mixtures.
| Τίτλος: | Quantitative TLC-based assay for the detection of urease inhibitors in complex mixtures. |
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| Συγγραφείς: | Cotacachi-Latacumba PE; Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Argentina. Electronic address: pcotacachi@fbioyf.unr.edu.ar., Salazar MO; Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Suipacha 531, Rosario, 2000, Argentina. Electronic address: msalazar@fbioyf.unr.edu.ar., Ramallo IA; Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Suipacha 531, Rosario, 2000, Argentina. Electronic address: aramallo@fbioyf.unr.edu.ar., Furlan RLE; Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Suipacha 531, Rosario, 2000, Argentina. Electronic address: rfurlan@fbioyf.unr.edu.ar. |
| Πηγή: | Analytica chimica acta [Anal Chim Acta] 2026 Nov 15; Vol. 1423, pp. 346178. Date of Electronic Publication: 2026 Aug 27. |
| Τύπος έκδοσης: | Journal Article |
| Γλώσσα: | English |
| Στοιχεία περιοδικού: | Publisher: Elsevier Country of Publication: Netherlands NLM ID: 0370534 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1873-4324 (Electronic) Linking ISSN: 00032670 NLM ISO Abbreviation: Anal Chim Acta Subsets: MEDLINE |
| Imprint Name(s): | Publication: Amsterdam : Elsevier Original Publication: Amsterdam. |
| Ιατρικοί όροι (MeSH): | Urease*/antagonists & inhibitors , Urease*/metabolism , Enzyme Inhibitors*/analysis , Enzyme Inhibitors*/pharmacology, Chromatography, Thin Layer/methods |
| Περίληψη: | Background: Urease inhibitors are increasingly relevant in agricultural and food chemistry because urease activity influences nitrogen-use efficiency, ammonia emissions, microbial survival, and food quality. Although numerous solution-based assays are available, no quantitative thin-layer chromatography (TLC)-based bioautographic method has been reported for detecting and localizing urease inhibitors directly within complex mixtures. This study aimed to develop and validate a quantitative TLC bioautographic assay suitable for effect-directed analysis of food-related matrices. Results: The assay combines chromatographic separation, urease immobilization in an agar layer, and ammonia detection through the Berthelot reaction to quantitatively visualize urease inhibition zones while avoiding false-positive responses associated with pH indicator-based detection. Validation showed linearity for thiourea over 0.47-15 μg/spot (R2 > 0.999), with LOD and LOQ values of 0.15 ± 0.08 and 0.46 ± 0.25 μg/spot, respectively, and intra-plate and intraday RSDs of 7.6-11.8% and 9.9%, respectively. Parallel screening of essential oils enabled rapid comparative evaluation of urease-inhibitory activity, while direct gas chromatography-mass spectrometry analysis of active chromatographic zones allowed microscale identification of eugenol as the principal bioactive constituent without preparative isolation. Significance: This work presents the first quantitative TLC-based bioautographic assay for urease inhibitors and establishes an integrated effect-directed analytical workflow that combines quantitative activity localization with direct molecular identification. The methodology provides a simple and cost-effective platform for screening and characterizing urease inhibitors in complex food- and agriculture-related matrices, facilitating the discovery of bioactive compounds with potential applications in food preservation, quality control, and sustainable agricultural practices. (Copyright © 2026 Elsevier B.V. All rights reserved.) |
| Competing Interests: | Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. |
| Contributed Indexing: | Keywords: Bioactivity-guided identification; Bioautography; Effect-directed analysis; Mass spectrometry; Natural products |
| Substance Nomenclature: | EC 3.5.1.5 (Urease) 0 (Enzyme Inhibitors) |
| Entry Date(s): | Date Created: 20261006 Date Completed: 20261006 Latest Revision: 20261006 |
| Update Code: | 20261007 |
| DOI: | 10.1016/j.aca.2026.346178 |
| PMID: | 42838678 |
| Βάση Δεδομένων: | MEDLINE |
| ISSN: | 1873-4324 |
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| DOI: | 10.1016/j.aca.2026.346178 |