Application of Optimized and Validated Agar Overlay TLC–Bioautography Assay for Detecting the Antimicrobial Metabolites of Pharmaceutical Interest
Abstract
The agar overlay TLC-bioautography is one of the crucial methods for simultaneous in situ detection and separation of antimicrobial metabolites of pharmaceutical interest. The main focus of this research relies on the dereplication of an antimicrobial metabolite coriloxin derived from mycoendophytic Xylaria sp. NBRTSB-20 with a validation of agar overlay TLC-bioautography technique. This polyketide metabolite coriloxin was purified by column chromatography, and its purity was assessed by HPLC, UPLC-ESI-QTOF-MS, FT-IR and NMR spectral analysis. The antimicrobial capability of ethyl acetate extract and the purified compound coriloxin was determined by disc diffusion, minimal inhibitory concentration and agar overlay TLC-bioautography assay. The visible LOD of coriloxin antimicrobial activity was found at 10 μg for Escherichia coli and 20 μg for both Staphylococcus aureus and Fusarium oxysporum. Inter- and intra-day precision was determined as the relative standard deviation is less than 6.56%, which proved that this method was precise. The accuracy was expressed as recovery, and the values were found ranging from 91.18 to 108.73% with RSD values 0.94-2.30%, respectively. The overall findings of this investigation suggest that agar overlay TLC-bioautography assay is a suitable and acceptable method for the in situ determination of antimicrobial pharmaceuticals.
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TLC-bioautography is highly effective and widely employed in natural product research for screening and assessment of bioactive compounds including antimicrobial agents.
“TLC-bioautography is a highly effective technique widely employed in natural product research for the screening and assessment of bioactive compounds including antimicrobial agents [77, 78].”
From Methods for screening and evaluation of antimicrobial activity: A review of protocols, advantages, and limitations (2024) · cites this paper as [77]
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Hossain, T. J. (2024) · European Journal of Microbiology and Immunology · doi:10.1556/1886.2024.00035 · Cited by 392 · see citing works
- Authors
- Nuthan, B. R., Rakshith, D., Marulasiddaswamy, K. M., Rao, H. C. Y., Ramesha, K. P., Mohana, N. C., Siddappa, S., Darshan, D., Kumara, K. K. S., & Satish, S.
- Affiliations
- University of Mysore (IN)
- Indian Institute of Science Bangalore (IN)
- Central Food Technological Research Institute (IN)
- Davangere University (IN)
- Corresponding author
- Satish, Sreedharamurthy
- ORCID
- Venue
- Journal of Chromatographic Science
- Publisher
- Oxford University Press (OUP)
- Published
- 2020-08-07
- Volume
- 58
- Issue
- 8
- Pages
- 737-746
- ISSN
- 0021-9665 (ISSN-L)
- 1945-239X
- OpenAlex
- W3048131226
- Cited by
- 25
- References
- 0
- Citations by year
- 2026: 3 · 2025: 3 · 2024: 3 · 2023: 4 · 2022: 4 · 2021: 6 · 2020: 2
- FWCI
- 2.08
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model (vor)
- Locations
- Journal of Chromatographic Science (publishedVersion)
- PubMed (publishedVersion)
- MyPrints@UOM (Mysore University Library) (acceptedVersion)
- NOT FOUND REPOSITORY (Indian Institute of Science Bangalore) (acceptedVersion)
- Funders
- Department of Science and Technology, Ministry of Science and Technology, India
- Indian Council of Medical Research
- University of Mysore
- Topics
- Physical Sciences › Chemistry › Analytical Chemistry › Chromatography in Natural Products
- Health Sciences › Medicine › Pharmacology › Microbial Natural Products and Biosynthesis
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical Methods in Pharmaceuticals
- Keywords
- antimicrobial metabolites, dereplication, Xylaria, polyketide metabolites, method validation, Escherichia coli, Staphylococcus aureus, Fusarium oxysporum, minimum inhibitory concentration, drug metabolites
- MeSH terms
- Agar, Anti-Bacterial Agents, Ascomycota, Bacteria, Biological Assay, Biological Products, Chromatography, High Pressure Liquid, Chromatography, Thin Layer, Fusarium, Reproducibility of Results, Linear Models, Limit of Detection, Endophytes, Polyketides
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