[Skip to Content](https://www.sigmaaldrich.com#main-content) [](https://www.sigmaaldrich.com/US/en) Products Cart0 USEN Products [Login / Register](https://www.sigmaaldrich.com/oidc-sign-in) [Order Lookup](https://www.sigmaaldrich.com/US/en/order-lookup) [Quick Order](https://www.sigmaaldrich.com/US/en/quick-order) Cart0 [Home](https://www.sigmaaldrich.com/US/en)[Applications](https://www.sigmaaldrich.com/US/en/applications)[Analytical Chemistry](https://www.sigmaaldrich.com/US/en/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.   * * * ## Related Products Slide 1 of 12 1 of 3 [ \ Supelco \ 1.05721 \ TLC plates, Silica gel 60](https://www.sigmaaldrich.com/US/en/product/mm/105721) Quick View [ \ Supelco \ 1.05715 \ TLC plates, Silica gel 60 F₂₅₄](https://www.sigmaaldrich.com/US/en/product/mm/105715) Quick View [ \ Supelco \ 1.16485 \ TLC plates, Silica gel 60 W F₂₅₄s](https://www.sigmaaldrich.com/US/en/product/mm/116485) Quick View [ \ Supelco \ 1.05802 \ LuxPlate™ TLC plates, Silica gel 60 F254](https://www.sigmaaldrich.com/US/en/product/mm/105802) Quick View [ \ Supelco \ 1.05570 \ TLC Silica gel 60 F254](https://www.sigmaaldrich.com/US/en/product/mm/105570) Quick View [ \ Supelco \ 1.05748 \ TLC plastic sheets, Silica gel 60](https://www.sigmaaldrich.com/US/en/product/mm/105748) Quick View [ \ Supelco \ 1.05582 \ TLC aluminum sheets, Silica gel 60 with concentrating zone 20 x 2.5 cm](https://www.sigmaaldrich.com/US/en/product/mm/105582) Quick View [ \ Supelco \ 1.13728 \ HPTLC plates, Silica gel 60 F₂₅₄ with concentrating zone 20 x 2.5 cm](https://www.sigmaaldrich.com/US/en/product/mm/113728) Quick View [ \ Supelco \ 1.13792 \ PLC plates, Silica gel 60 F₂₅₄, 1 mm with concentrating zone 20 x 4 cm](https://www.sigmaaldrich.com/US/en/product/mm/113792) Quick View [ \ Supelco \ 1.05635 \ HPTLC plates, Silica gel 60 F₂₅₄ Multiformat pre-scored to 5 x 5 cm](https://www.sigmaaldrich.com/US/en/product/mm/105635) Quick View [ \ Supelco \ 1.05586 \ LiChrospher® HPTLC aluminum sheets, Silica gel 60 F₂₅₄s](https://www.sigmaaldrich.com/US/en/product/mm/105586) Quick View [ \ Supelco \ 1.05646 \ LiChrospher® HPTLC plates, Silica gel 60 RP-18 WF₂₅₄s](https://www.sigmaaldrich.com/US/en/product/mm/105646) Quick View * * * ## Featured Categories [](https://www.sigmaaldrich.com/US/en/products/analytical-chemistry/analytical-chromatography/tlc-plates-and-adsorbants) [TLC Plates & Adsorbants](https://www.sigmaaldrich.com/US/en/products/analytical-chemistry/analytical-chromatography/tlc-plates-and-adsorbants) Choose from classical TLC plates with silica, aluminum, glass, and plastic backings as well as HPTLC plates, MS-Grade TLC and HPTLC plates, and PLC plates. [Shop Products](https://www.sigmaaldrich.com/US/en/products/analytical-chemistry/analytical-chromatography/tlc-plates-and-adsorbants) [](https://www.sigmaaldrich.com/US/en/products/analytical-chemistry/analytical-chromatography/hplc-columns) [HPLC Columns](https://www.sigmaaldrich.com/US/en/products/analytical-chemistry/analytical-chromatography/hplc-columns) Achieve precise separations with our extensive HPLC column collection. Enhance retention, resolution, and selectivity. Order today.   [Shop Products](https://www.sigmaaldrich.com/US/en/products/analytical-chemistry/analytical-chromatography/hplc-columns) [](https://www.sigmaaldrich.com/US/en/products/analytical-chemistry/analytical-chromatography/solvents) [Solvents](https://www.sigmaaldrich.com/US/en/products/analytical-chemistry/analytical-chromatography/solvents) Your Solvent Source: Find the right fit with Supelco®, SigmaAldrich®, & SAFC® brands, covering analytical, lab, & biopharmaceutical uses. Order online.   [Shop Products](https://www.sigmaaldrich.com/US/en/products/analytical-chemistry/analytical-chromatography/solvents) [](https://www.sigmaaldrich.com/US/en/products/chemistry-and-biochemicals/lab-chemicals/acids) [Acids](https://www.sigmaaldrich.com/US/en/products/chemistry-and-biochemicals/lab-chemicals/acids) Uncover a wide acid range: Supelco® for analysis, Sigma-Aldrich® for labs, SAFC® for biopharma. Tailor solutions for varied needs.   [Shop Products](https://www.sigmaaldrich.com/US/en/products/chemistry-and-biochemicals/lab-chemicals/acids) * * * Overview Related Articles & Protocols Support 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. ## [](https://www.sigmaaldrich.com)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.  ## [](https://www.sigmaaldrich.com)TLC Retention Factor (Rf) The retention factor (Rf) is used to measure the movement of compounds along the TLC plate. Rf 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. | | | |------|--------------------------------| | Rf = | distance traveled by component | | | 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 Rf values, whereas polar compounds tend to move slowly and have lower Rf values. ## [](https://www.sigmaaldrich.com)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 [](https://www.sigmaaldrich.com/documents-search) [Looking for More Specific Information?](https://www.sigmaaldrich.com/documents-search) Visit our document search for data sheets, certificates and technical documentation. [Find Documents](https://www.sigmaaldrich.com/documents-search) ## Related Articles - [High-Performance Thin-Layer Chromatography: A Fast and Efficient Fingerprint Analysis Method for Medicinal Plants](https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/analytical-chemistry/thin-layer-chromatography/high-performance-thin-layer-chromatography) In this article we present several HPTLC applications and analytical standards for ginsenosides. - [History of Thin-Layer Chromatography](https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/analytical-chemistry/thin-layer-chromatography/tlc-history) Discover the history of thin-layer chromatography (TLC) and its key milestones that have shaped analytical chemistry over the past century. - [Determination of Stevioside and Rebaudioside A in Stevia leaves, beverages and food additives using HPTLC-MS](https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/analytical-chemistry/thin-layer-chromatography/tlc-ms-determination-of-stevioside-and-rebaudioside) HPTLC-MS was used to analyze stevioside and rebaudioside in artificial sweeteners, stevia plants, cola and isotonic drinks with a minimum of sample preparation. - [TLC Plate Selection Guide](https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/analytical-chemistry/thin-layer-chromatography/tlc-plate-selection) Thin layer chromatography (TLC), an analytical technique often used to separate and identify compounds present in a given mixture, can also be used to determine the purity of a particular substance within that mixture - [TLC Silica Gel 60 G Plates with Gypsum](https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/analytical-chemistry/thin-layer-chromatography/tlc-silica-gel-60g-plates-with-gypsum) TLC plates, with silica gel, gypsum as binder, International Pharmacopoeia compliant, for identification of Atenolol and Chlorthalidone in tablets. - [See All (19)](https://www.sigmaaldrich.com/US/en/search/facet-search?focus=sitecontent&term=facet-search) ## Related Protocols - [Thin-Layer Chromatography Process](https://www.sigmaaldrich.com/US/en/technical-documents/protocol/analytical-chemistry/thin-layer-chromatography/tlc-process) TLC separation relies on capillary forces to transport substances on a stationary phase after mobile phase transport. - [Chromatographic Testing of Black Pepper](https://www.sigmaaldrich.com/US/en/technical-documents/protocol/analytical-chemistry/thin-layer-chromatography/chromatographic-testing-of-black-pepper-according-to-the-usp) Powdered black pepper tested for pierine content follows USP guidelines using HPTLC and HPLC for dietary supplements. - [Determination of Caffeine in Coffee Using High Performance Thin Layer Chromatography](https://www.sigmaaldrich.com/US/en/technical-documents/protocol/analytical-chemistry/thin-layer-chromatography/quantification-of-caffeine) HPTLC was used to analyze caffeine in coffee. Filtration of coffee through a PTFE syringe filter was the only sample preparation. Quantitation was conducted with a TLC scanner. - [Thin Layer Chromatography Identification Test for Dextromethorphan following European Pharmacopeia Monograph on Silica Gel G 254 Plates](https://www.sigmaaldrich.com/US/en/technical-documents/protocol/analytical-chemistry/thin-layer-chromatography/thin-layer-chromatography-identification-test-for-dextromethorph) 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 Shampoos by HPTLC](https://www.sigmaaldrich.com/US/en/technical-documents/protocol/analytical-chemistry/thin-layer-chromatography/caffeine-taurine-and-arginine-in-shampoos-by-hptlc) The modern thin-layer chromatography (TLC) offers advantages also in the field of cosmetics to quickly and simultaneously analyze samples with densitometric detection. - [See All (13)](https://www.sigmaaldrich.com/US/en/search/facet-search?focus=sitecontent&term=facet-search) ### Find More Articles and Protocols Enter Keywords Search ## How Can We Help In case of any questions, please submit a [customer support request](https://www.sigmaaldrich.com/US/en/support/customer-support) or talk to our customer service team: Email [custserv@sial.com](mailto:custserv@sial.com) or call +1 (800) 244-1173 ## Additional Support - [Chromatogram Search](https://www.sigmaaldrich.com/chromatogram-search) Use the Chromatogram Search to identify unknown compounds in your sample. - [Calculators & Apps](https://www.sigmaaldrich.com/US/en/support/calculators-and-apps) Web Toolbox - science research tools and resources for analytical chemistry, life science, chemical synthesis and materials science. - [Customer Support Request](https://www.sigmaaldrich.com/US/en/support/customer-support) Customer support including help with orders, products, accounts, and website technical issues. - [FAQ](https://maestro.my.site.com/knowledgeportal/s/) Explore our Frequently Asked Questions for answers to commonly asked questions about our products and services. * * * __Featured Articles__ [High-Performance Thin-Layer Chromatography: A Fast and Efficient Fingerprint Analysis Method for Medicinal Plants](https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/analytical-chemistry/thin-layer-chromatography/high-performance-thin-layer-chromatography) In this article we present several HPTLC applications and analytical standards for ginsenosides. [Determination of Stevioside and Rebaudioside A in Stevia leaves, beverages and food additives using HPTLC-MS](https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/analytical-chemistry/thin-layer-chromatography/tlc-ms-determination-of-stevioside-and-rebaudioside) HPTLC-MS was used to analyze stevioside and rebaudioside in artificial sweeteners, stevia plants, cola and isotonic drinks with a minimum of sample preparation. [TLC Plate Selection Guide](https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/analytical-chemistry/thin-layer-chromatography/tlc-plate-selection) Thin layer chromatography (TLC), an analytical technique often used to separate and identify compounds present in a given mixture, can also be used to determine the purity of a particular substance within that mixture [TLC Silica Gel 60 G Plates with Gypsum](https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/analytical-chemistry/thin-layer-chromatography/tlc-silica-gel-60g-plates-with-gypsum) TLC plates, with silica gel, gypsum as binder, International Pharmacopoeia compliant, for identification of Atenolol and Chlorthalidone in tablets. Top __Sign In To Continue__ To continue reading please sign in or create an account. Sign In__Don't Have An Account?__Register An unknown error has occured. - English - EN - Español - ES [Learn More](https://www.sigmaaldrich.com)