Thin Layer Chromatography (TLC)

Thin layer chromatography (TLC) is a method that uses a thin layer made of a stationary phase such as silica gel, polyamide, alumina, etc. on a glass or aluminum plate, separates the mixture sample into each component with a mobile phase. TLC is widely used for substance identification, purity testing, and for examining separation conditions when using column chromatography. Fujifilm Wako provides TLC plates for simple analysis, preparative separation and purification, coated with various stationary phases according to the purpose. Fujifilm Wako also offers various related products such as carriers for TLC plate preparation and ready-made TLC stains.

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How to Select TLC Plate

(1) Type of carrier

Unmodified silica gel	Used in adsorption chromatography mode, mainly suitable for separation of acidic or neutral compound.
Chemically modified silica gel	Various functional groups are bonded to the silanol groups on the surface of unmodified silica gel to change the function. C18 modified silica gel, which is octadecyl group (ODS) bonded silica gel, suitable for reversed-phase chromatography mode. NH₂ modified silica gel, which is aminopropyl group bonded silica gel, suitable for separation of sugars, catecholamines, etc.
Polyamides	Used in adsorption chromatography mode, suitable for separation of phenols and carboxylic acids.
Alumina	Used in adsorption chromatography mode, suitable for separation of basic compounds.
Cellulose	Used in Normal phase chromatography mode, suitable for separation of hydrocarbon compounds.

(2) Pore size/specific surface area

The smaller the pore size, the greater the specific surface area of the silica gel and the higher the resolution of small molecules. Conversely for macromolecules such as proteins, the larger the pore size, the higher the resolution. It is necessary to select an appropriate pore size so that sample molecules can easily enter and exit the pores.

(3) Type of support

Glass has excellent chemical resistance and is the most widely used. Aluminum and paper can be cut easily with scissors, but the carrier peels off from support or the TLC bends depending on the developing solvent. In addition, strong acids/bases cannot be used as developing solvents or coloring reagents.

(4) Type of fluorescent substance

Not contain	It is suitable for detection of fluorescing samples and spot detection using coloring TLC stains (sulfuric acid, ninhydrin, iodine, etc.) because it does not contain fluorescent substances.
Mono fluorescent substance	254 nm UV irradiation causes the entire plate to fluoresce green, and samples with UV absorption turn black colored spots.
Mixture fluorescent substance	Red, green and blue fluorophores substance are added. Under broadband UV (250-400 nm) irradiation causes the entire plate to white, and samples with UV absorption turn the characteristic colored spot such as, red, blue, etc.

Physical Properties of TLC Plates

Silica gel (unmodified)

Product name	Pore size	Specific surface area	Pore volume	Layer thickness	Support	Sticking agent	Fluorescent substance	Detection
Silicagel 70 TLC Plate-Wako	7 nm	450 m²/g	0.8 mL/g	190-290 μm	Glass plate	Polymer	Not contain	Color TLC stains
Silicagel 70 F₂₅₄ PLC Plate-Wako							Mono fluorescent substance	UV (254 nm)
Silicagel 70 FM TLC Plate-Wako							Mixture fluorescent substance	Broad UV (250-400 nm)
Silicagel 70 PF₂₅₄ Plate-Wako				700-900 μm			Mono fluorescent substance	UV (254 nm)
Silicagel 70 F₂₅₄ TLC Plate-Wako				900-1200 µm			Mono fluorescent substance	UV (254 nm)
Wakogel^®FM Plate				180-250 μm		Cast/starch	Mixture fluorescent substance	Broad UV (250-400 nm)
Chromato Sheet	-	-	-	300-400 μm	Paper	-	Mono fluorescent substance	UV (254 nm)

NH₂ modified silica gel

Product name	Pore size	Specific surface area	Pore volume	Layer thickness	Support	Sticking agent	Fluorescent substance	Detection
NH₂ Silica Gel 60 F₂₅₄ Plate-Wako (layer thickness 0.25 mm)	6 nm	450 m²/g	0.7 mL/g	190-250 μm	Grass plate	Polymer	Mono fluorescent substance	UV (254 nm)
NH₂ Silica Gel 60 F₂₅₄ Plate-Wako (layer thickness 0.5 mm)				400-550 μm
NH₂ Silica Gel 60 F₂₅₄ Plate-Wako (layer thickness 0.75 mm)				650- 850 μm

Polyamide

Product name	Pore size	Specific surface area	Pore volume	Layer thickness	Support	Sticking agent	Fluorescent substance	Detection
Polyamide FM Plate	-	-	-	70-110 µm	Glass plate	Cast	Mixture fluorescent substance	Broad UV (250-400 nm)

TLC Plate Outer Box

- Outer box made of paper for easy disposal.
- Also can be used for storage!
- Smart design without bulk (vertical 234 mm × width 226 mm × thickness 58 mm).

How to use

What is Thin Layer Chromatography (TLC)?

Thin-Layer Chromatography (TLC) is chromatography using a thin layer plate in which a carrier is fixed to a support. When one end of the TLC is immersed in solvent, the solvent moves upward through the gaps in the carrier. If sample is present on the plate, the sample will move by the solvent movement. At this time, the distance that the sample moves (Rf value) varies depending on the degree of adsorption to the carrier and the difference/balance in affinity to the moving solvent (mobile phase). From this principle, Rf value is used to separate and identify organic compounds in TLC.

Author: Y. Suto

For research use or further manufacturing use only. Not for use in diagnostic procedures.

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