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Thin-Layer Chromatography (TLC): Overview

Thin-Layer Chromatography (TLC): Overview

Thin-layer chromatography (TLC) is a chromatography technique that separates compounds based on their polarity. TLC typically uses polar silica gel, a form of silicon dioxide, as the stationary phase. The silica gel contains hydroxyl (OH) groups on its surface, which form hydrogen bonds with polar compounds, influencing their adhesion to the stationary phase.
To begin the analysis, a mixture of compounds is spotted on the starting line on the TLC plate using a thin capillary. The bottom of the...

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Related Experiment Video

Updated: Jun 13, 2026

Thin-layer Chromatographic TLC Separations and Bioassays of Plant Extracts to Identify Antimicrobial Compounds

Thin-layer Chromatographic TLC Separations and Bioassays of Plant Extracts to Identify Antimicrobial Compounds

Published on: March 27, 2014

Thin Layer Chromatography Bioautography Scanning Electron Microscopy (TLCBA SEM) method for antimicrobial fraction

Anna du Toit¹, Chaitali Dekiwadia¹, Lisa Cornell¹

¹School of Science & Microscopy & Microanalysis Facility, Royal Melbourne Institute of Technology University, Melbourne, Australia.

Journal of Microbiological Methods

|August 29, 2025

Summary

Thin Layer Chromatography Bio-autography combined with Scanning Electron Microscopy offers a faster method to assess antibacterial effects on bacterial morphology. This technique efficiently evaluates anti-staphylococcal compounds, reducing compound and time requirements.

Keywords:

Antibacterial Scanning electron microscopy Structure damage Target Thin layer chromatography-bioautography

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Last Updated: Jun 13, 2026

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Published on: March 27, 2014

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Area of Science:

Microbiology
Analytical Chemistry

Background:

Bacterial morphology is crucial for understanding infection and treatment response.
Traditional methods for assessing antibacterial activity can be time-consuming and require substantial test compounds.

Purpose of the Study:

To develop and validate a combined Thin Layer Chromatography Bio-autography and Scanning Electron Microscopy (TLCBA-SEM) method.
To evaluate the effect of chromatographed fractions on bacterial morphology efficiently.

Main Methods:

Thin Layer Chromatography Bio-autography (TLCBA) was coupled with Scanning Electron Microscopy (SEM).
Bacterial morphology was observed directly on coverslips treated with specific chromatographic fractions.
Fraction CS1, known for anti-staphylococcal activity, was tested using the developed method.

Main Results:

The TLCBA-SEM method allowed direct observation of bacterial morphology changes induced by chromatographic fractions.
Coverslips treated with the anti-staphylococcal fraction CS1 yielded results comparable to traditional plate-dilution methods.
The combined technique demonstrated reduced requirements for test compounds and processing time.

Conclusions:

TLCBA-SEM is an effective and efficient approach for evaluating the impact of bioactive compounds on bacterial morphology.
This method offers a valuable alternative to conventional techniques, saving time and resources in antibacterial drug discovery.