2/3/24, 10:28 AM Thin Layer Chromatography

Type in Product Names, Product Numbers, or CAS Numbers to see suggestions.

Home Applications Analytical Chemistry Thin Layer Chromatography

Thin Layer Chromatography

Thin layer chromatography (TLC) is an affinity-based method used to separate compounds in a mixture. TLC is a highly versatile separation method
that is widely used for both qualitative and quantitative sample analysis. TLC can be used to analyze virtually any substance class, including pesticides,
steroids, alkaloids, lipids, nucleotides, glycosides, carbohydrates, and fatty acids.

In TLC, the stationary phase is a thin adsorbent material layer, usually silica gel or aluminum oxide, coated onto an inert plate surface, typically glass,
plastic, or aluminum. The sample is spotted onto one end of the TLC plate and placed vertically into a closed chamber with an organic solvent (mobile
phase). The mobile phase travels up the plate by capillary forces and sample components migrate varying distances based on their differential
affinities for the stationary and mobile phases. When the solvent reaches the top of the plate, the plate is removed from the developing chamber and
dried. The separated components appear as spots on the plate and the retention factor (Rf) of each component is assessed.

TLC Process and Principles

TLC Retention Factor: R f
Applications of TLC
Feedback

FIND CHROMATOGRAMS
Chromatogram Search

TOP

RELATED TECHNICAL ARTICLES RELATED PROTOCOLS

High Performance Thin Layer Chromatography: A Fast and Efficient Thin Layer Chromatography Process
https://www.sigmaaldrich.com/BD/en/applications/analytical-chemistry/thin-layer-chromatography?psafe_param=1&utm_source=google&utm_me… 1/3
2/3/24, 10:28 AM Thin Layer Chromatography
High-Performance Thin-Layer Chromatography: A Fast and Efficient… Thin-Layer Chromatography Process

In this article we present several HPTLC applications and analytical standards What causes a dissolved mixture to separate when applied to a TLC plate? The
for ginsenosides. thin-layer chromatography process relies on capillary forces. During…

History of Thin-Layer Chromatography
Thin layer chromatography
Determination of Stevioside and Rebaudioside A in Stevia leaves,…
HPTLC-MS was used to analyze stevioside and rebaudioside in artificial
sweeteners, stevia plants, cola and isotonic drinks with a minimum of sample…
TLC Plate Selection Guide
Thin layer chromatography (TLC), an analytical technique often used to
separate and identify compounds present in a given mixture, can also be use…
TLC Silica Gel 60 G Plates with Gypsum
TLC plates, with silica gel, gypsum as binder, International Pharmacopoeia
compliant, for identification of Atenolol and Chlorthalidone in tablets
See All (9)
Chromatographic Testing of Black Pepper According to the United States…
Powdered black pepper was chosen as example for dietary supplements, being
a source for pierine. It is tested according to the current United States…
Determination of Caffeine in Coffee using High Performance Thin Layer…
HPTLC was used to analyze caffeine in coffee. Filtration of coffee through a
PTFE syringe filter was the only sample preparation. Quantitation was…
Thin Layer Chromatography Identification Test for Dextromethorphan…
Silica gel G 254 plates are suitable for analysis of Dextromethorphan following
the European pharmacopeia monograph.
Detection and Determination of Caffeine, Taurine and Arginine in…
The modern thin-layer chromatography (TLC) offers advantages also in the
field of cosmetics to quickly and simultaneously analyze samples with…
See All (13)

Find More Articles and Protocols

Enter Keywords

Search

TLC PROCESS AND PRINCIPLES

TLC is based on the classic chromatography principle where mixture components are separated between a fixed stationary phase and a liquid mobile
phase by differential affinities between the two phases.

TLC RETENTION FACTOR (R f )

The retention factor (R f ) is used to measure the movement of compounds along the TLC plate. R f is defined as the distance travelled by an individual
component divided by the total distance travelled by the solvent. Its value is always between zero and one.

distance traveled by component

Rf =
distance traveled by solvent

In general, the stronger a compound binds to the stationary phase adsorbent, the slower it migrates up the TLC plate. As TLC adsorbents are typically
polar, non-polar compounds tend to travel more rapidly up the plate, resulting in a higher R f values, whereas polar compounds tend to move slowly
and have lower R f values.

APPLICATIONS OF TLC
TLC is widely used by many industries and research fields, including pharmaceuticals, clinical testing, environmental toxicology, food, water and
pesticide analysis, and cosmetics. Typical applications of TLC include:

Analysis of drug residues and antibiotics in food and environmental samples

Identification and quantification of colors, ingredients, preservatives, and sweetening agents in food and cosmetic products
Quality control and purity testing of pharmaceutical formulations
Rapid, high-throughput screening prior to HPLC
Examination of chemical reactions for completion

RELATED APPLICATIONS
Large Molecule HPLC
High performance liquid chromatography (HPLC) can be used to separate and identify different large biomolecules such as protein and peptides in a sample. It is based on
the pumping of a sample with a solvent (mobile phase) through a column packed with sorbent material (stationary phase) at a high pressure.

Gas Chromatography (GC)

Gas chromatography is a common analytic technique used to separate and analyze volatile compounds in the gas phase. GC is applied in many industries for quality control,
and to identify and/or quantify compounds in a mixture.

Mass Spectrometry
https://www.sigmaaldrich.com/BD/en/applications/analytical-chemistry/thin-layer-chromatography?psafe_param=1&utm_source=google&utm_me… 2/3
2/3/24, 10:28 AM Thin Layer Chromatography
Mass Spectrometry

Mass spectrometry (MS) is an analytical tool used to identify compounds, determine chemical structure, and assess isotopic abundance. In MS, samples are ionized, and the
resulting ions are identified based on their mass-to-charge (m/z) ratios.

Wet Chemical Analysis

The wet chemical analysis involves identifying and quantifying the desired elements present in a liquid sample using several measuring techniques. With chemical reagents,
the analyte converts a dye proportionally, which can be read-out visually or photometrically.

Low Pressure Liquid Chromatography

Low pressure liquid chromatography (LPLC) is a chromatographic technique that operates at low pressures to drive the mobile phase onto the column containing a
stationary phase by the action of a pump.

Small Molecule HPLC

Small molecules are ions and compounds of molecular weight typically less than 900 daltons. These compounds can be effectively separated and analyzed by HPLC, UHPLC
and LC-MS using mainly silica particles or monolithic stationary phases with a broad range of column chemistries (modifications).

RELATED PRODUCTS

TLC plates, Silica gel 60

pkg of 25 plates, plate L × W 20 cm × 20 cm, glass support

RELATED PRODUCT CATEGORIES

Thin Layer Chromatography (TLC)
We offer a variety of TLC plates including classical TLC plates with silica, aluminum, glass, and plastic backings in various dimensions as per your needs. Also available are
the HPTLC plates, MS-Grade TLC and HPTLC plates, and PLC plates.

LPLC Media & Accessories

Browse through our portfolio to choose from a wide selection of products required for LPLC applications such as columns, a variety of adsorbents, ion exchange resins,
separation media, and accessories, designed to fit your unique application demands.

HPLC Columns
Our wide selection of HPLC columns for small and large molecule analysis allows you to enhance the separation process in terms of retention, resolution, selectivity, and
analysis time, all while reducing the time for method development.

Solvents
https://www.sigmaaldrich.com/BD/en/applications/analytical-chemistry/thin-layer-chromatography?psafe_param=1&utm_source=google&utm_me… 3/3