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Thin-Layer Chromatography (TLC): Overview01:11

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...
Chromatographic Methods: Terminology01:18

Chromatographic Methods: Terminology

Chromatography is an analytical technique widely used in fields such as chemistry, biology, environmental science, and pharmaceuticals to separate the components of a mixture and identify substances between them. The process of chromatography is based on the interactions between two distinct phases: the stationary phase and the mobile phase. The stationary phase is fixed in place by a supporting material, while the mobile phase moves over it, carrying the solutes. As the mobile phase travels,...
Differential Staining Technique01:26

Differential Staining Technique

Differential staining is an essential microbiological technique that exploits variations in cell wall structures to classify and identify microorganisms. It facilitates the distinction of bacteria, aiding in diagnostic and research applications. Two of the most widely used differential staining methods are Gram staining and acid-fast staining, both of which rely on the chemical and structural differences in bacterial cell walls.Gram Staining TechniqueGram staining differentiates bacteria by...
Fixation and Sectioning01:03

Fixation and Sectioning

Two basic types of preparation are used to visualize specimens with a light microscope: wet mounts and fixed specimens.
The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...
Chromatographic Methods: Classification01:12

Chromatographic Methods: Classification

Chromatographic techniques are classified in three ways: the classification is based on the physical state of the stationary and mobile phases, how the mobile phase and the stationary phase contact each other, or through the chemical or physical processes that isolate the components of the sample. Typically, the mobile phase is either a liquid or gas, while the stationary phase is either a solid or a liquid layer applied to a solid surface.
Chromatographic techniques are typically named by...
Simple Staining Technique01:24

Simple Staining Technique

OverviewStaining techniques in microscopy enhance the visualization of microorganisms by increasing contrast and allowing the differentiation of cellular structures. Simple staining is one of the fundamental methods used to observe the basic morphological characteristics of microorganisms, including their size, shape, and arrangement. This method relies on the application of a single dye to stain the entire cell, producing a clear contrast between the cell and the background.FixationFixation is...

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Related Experiment Video

Updated: Jul 13, 2026

Estimating the Yield of Compounds on the TLC Plate via the Blue-LED Illumination Technique
06:14

Estimating the Yield of Compounds on the TLC Plate via the Blue-LED Illumination Technique

Published on: October 6, 2022

Evaluation of quantitative thin layer chromatography using staining reagents.

Richard Johnsson1, Gustav Träff, Martin Sundén

  • 1Organic Chemistry, Lund University, SE-221 00, Lund, Sweden.

Journal of Chromatography. A
|August 7, 2007
PubMed
Summary

Quantitative thin-layer chromatography (TLC) using staining reagents offers reliable analysis for organic compounds lacking chromophores. Modern image analysis enhances accuracy, making it suitable for high-throughput screening and physical organic chemistry research.

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

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Arabidopsis thaliana Polar Glycerolipid Profiling by Thin Layer Chromatography (TLC) Coupled with Gas-Liquid Chromatography (GLC)
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Arabidopsis thaliana Polar Glycerolipid Profiling by Thin Layer Chromatography (TLC) Coupled with Gas-Liquid Chromatography (GLC)

Published on: March 18, 2011

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Last Updated: Jul 13, 2026

Estimating the Yield of Compounds on the TLC Plate via the Blue-LED Illumination Technique
06:14

Estimating the Yield of Compounds on the TLC Plate via the Blue-LED Illumination Technique

Published on: October 6, 2022

Thin-layer Chromatographic (TLC) Separations and Bioassays of Plant Extracts to Identify Antimicrobial Compounds
12:04

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

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Arabidopsis thaliana Polar Glycerolipid Profiling by Thin Layer Chromatography (TLC) Coupled with Gas-Liquid Chromatography (GLC)
13:02

Arabidopsis thaliana Polar Glycerolipid Profiling by Thin Layer Chromatography (TLC) Coupled with Gas-Liquid Chromatography (GLC)

Published on: March 18, 2011

Area of Science:

  • Analytical Chemistry
  • Organic Chemistry
  • Chromatography

Background:

  • Thin-layer chromatography (TLC) is crucial for analyzing organic compounds, especially those without chromophores.
  • Traditional TLC methods can be limited in quantitative accuracy.

Purpose of the Study:

  • To investigate the quantitative capabilities of TLC using staining reagents combined with modern image analysis.
  • To assess the reliability and error sources in quantitative TLC for various applications.

Main Methods:

  • Quantitative thin-layer chromatography (TLC) utilizing staining reagents.
  • Application of modern image analysis software for data processing.
  • Analysis of error ranges and detection limits throughout the process.

Main Results:

  • Demonstrated the feasibility of obtaining reliable quantitative measurements using stained TLC.
  • Identified the staining process as a primary source of error.
  • Showed that measuring compound ratios can significantly diminish errors.

Conclusions:

  • Quantitative TLC with staining reagents and image analysis is a viable method for analyzing non-chromophoric organic compounds.
  • The technique is suitable for high-throughput screening and physical organic chemistry.
  • Error reduction strategies, such as ratio measurements, enhance quantitative accuracy.