Thin Layer Chromatography - Sorbent Technologies · PDF fileThin Layer Chromatography Sorbent...

112www.sorbtech.com 770-936-0323 Sorbent Technologies, Inc.

Thin Layer ChromatographySorbtech offers all chemistries currently possible to manufacture in a TLC format – silica gel, bonded silica gel, alumina, cellulose, and so on - in all sizes ranging from microscope slide sized to full 20cm x 20cm plates.

• TLC Plates

• TLC Plate Accessories

• TLC Glass Plate Cutter

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TLC Plates

Thin Layer ChromatographySorbent Technologies offers the highest performance and widest selection of TLC plates on glass, plastic, and aluminum supports. Analytical and preparative layers with different sorbent coatings: silica gel, alumina, cellulose, bonded phases, polyamide, chiral, and more are available. TLC plates are available in a wide array of sizes from 20 x 20 cm to 2.5 x 7.5 cm with or without UV254 indicator. HPTLC plates provide higher performance and better resolution. Inquire about custom plates. We also carry TLC accessories.

QualityOur TLC plates are guaranteed by stringent production control including standardized lot tests, surface checks for roughness or cracks as well as hardness and adherence checks. Plates and sheets are immediately ready for chromatographic separation and require no coatings or impregnations. TLC layers are smooth, homogenous and well-adhering to produce reproducible quantitative evaluations.

TLC/HPTLC Advantages:• High sample throughput in a short time• Suitable for screening tests • Pilot Procedure for HPLC and flash chromatography• After separation the analytical information can be stored for a longer period of time (the TLC plate acts as a storage medium for data)• Separated substances can be subjected to subsequent analytical procedures (e.g. IR, MS) at a later date.• Rapid and cost effective optimization of the separation due to easy change of mobile and stationary phase.

TLC Plate and Sheet Characteristics:• Classical Adsorbents - for ~80% of all TLC separations silica 60 (mean pore diameter of 60 Å = 6 nm) is used. Other classical adsorbents are aluminium oxide, cellulose, diatomaceous earth, ion exchangers and polyamide. • Special Phases - reversed phases, mainly C18 (octadecyl) modified silica, but also cyano-, amino-, diol and C2 modified silica are available. Special layers for specific separations, like our Chiral TLC plate for enantiometer separation complete the versatile range of TLC plates. • Particle Size Distribution and Thickness of Layer – Particle size distribution and thickness of layer are chosen to fit the given type of application (e.g. HPTLC, standard or preventative separations). Most ready-to-use layers are available with or without fluorescent indicator.

TLC Plates Standardized Conditioning MethodThe following conditioning procedure allows proper standardization of TLC plate performance for reproducibility from chemist to chemist and plate to plate.1. Pre-wash your plate to remove impurities: this will result in a more reproducible separation and purer extracted components.

a. General pre-washing: use methanol or methanol/chloroform (1:1). Develop overnight to top of plate. Let air dry in a ventilation hood.b. Extraction pre-washing: use the solvent system that you are going to use for the extraction of the fractions after separation. c. Develop overnight to top of plate. Let air dry in a ventilation hood.

2. Activation strengthens the layer and increases reproducibility.d. Dry in a vacuum oven at 110° C for an houre. Let the plates cool under vacuum until room temperaturef. Store in a desiccator until use

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Cutting TipsFlexible Plastic and Aluminum Backed Plates:1. Keep your plates as dry as possible, storing in a desiccator if possible (if not in a desiccator, keep the box closed at all times).2. Using very sharp scissors, cut at a 45° angle leaning to the outside (therefore, if you are right handed, the cut should lean to the right and from the back)Glass Backed Plates:1. Buy the Sorbtech TLC Plate Glass Cutter (Cat.# TLC-Cutter) to enable you to optimize plate cutting for analytical and prep TLC plates. This will save both time and money from material waste and labor.

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Cat. # Layer Backing Thickness Indicator Size (cm) Qty/pkg4115026 Silica XHL glass 250 μm - 20 x 20 504115056 Silica XHL glass 250 μm - 10 x 10 254115067 Silica XHL glass 250 μm - 5 x 10 254115126 Silica XHL glass 250 μm UV254 20 x 20 254115126PS Silica XHL -Pre-Scored glass 250 μm UV254 20 x 20 254115137 Silica XHL glass 250 μm UV254 10 x 20 504115137PS Silica XHL -Pre-Scored glass 250 μm UV254 10 x 20 504115148 Silica XHL glass 250 μm UV254 5 x 20 1004115156 Silica XHL glass 250 μm UV254 10 x 10 254115167 Silica XHL glass 250 μm UV254 5 x 10 504115198 Silica XHL glass 250 μm UV254 2.5 x 7.5 1004214137 Nano-Silica XHL glass 200 μm UV254 10 x 20 504214156 Nano-Silica XHL glass 200 μm UV254 10 x 10 254214178 Nano-Silica XHL glass 200 μm UV254 5 x 5 100

Extra Hard Layers - Silica XHL TLCSilica Gel Extra Hard Layer (XHL) plates feature a newly developed binder system, resulting in an outstanding hardness of the silica layer. Due to its excellent abrasion resistance, the formation of dust on the surface of XHL Silica is minimized.

Advantages of the Silica XHL TLC: • The plate can be easily labeled with a lead pencil for clear identification. • The high abrasion resistance prevents undesired changes or damage to the layer (thickness) during transportation or handling. • A dust-free plate allows convenient handling and avoids cross-contamination of the TLC chamber. • The UV indicator of the XHL Silica features an increased detection sensitivity. • The increased sensitivity allows for successful applications in trace analysis.

Compared to Hard Layer (HL) and Soft Layer Silica G plates, Extra Hard Layer (XHL) Silica demonstrates improved separation efficiency, since the particle size distribution of the silica is optimized. The selectivity of XHL Silica is different compared to HL and Silica G. Thus the chromatographer now can choose from 3 different types of Silica plates (XHL, HL, and Silica G) in order to optimize a given chromatographic experiment.

Additionally a Nano-Silica XHL HPTLC plate is available with the advantage of higher resolution and shorter development time due to a finer particle distribution.

For more selections and sizes, visit our website or contact your personal Product Specialist.

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Soft Layers - Silica XG TLCSilica XG layers have a newly developed binding agent that makes it adhere to the aluminum substrate more strongly than the Silica G layers. This equates to less flaking of the coating.

For these TLC plates a silica gel with a 60 Å mean pore diameter, a 500 m2/g specific surface area, a 0.75 ml/g specific pore volume, and a particle size of 5-17 μm is used. Fluorescent indicator (UV254) and pre-cut sizes are available.

Silica XG: This is a soft layer plate that contains Gypsum.

Cat. # Layer Backing Thickness Indicator Size (cm) Qty/pkg2115126 Silica HL glass 250 μm UV254 20 x 20 252115137 Silica HL glass 250 μm UV254 10 x 20 502115148 Silica HL glass 250 μm UV254 5 x 20 1002115169 Silica HL glass 250 μm UV254 5 x 10 2002115167 Silica HL glass 250 μm UV254 5 x 10 502115026 Silica HL glass 250 μm - 20 x 20 252115048 Silica HL glass 250 μm - 5 x 20 1002115037 Silica HL glass 250 μm - 10 x 20 502115067 Silica HL glass 250 μm - 5 x 10 50

Cat. # Layer Backing Thickness Indicator Size (cm) Qty/pkg4434126 Silica XG aluminum 200 μm UV254 20 x 20 254434135 Silica XG aluminum 200 μm UV254 10 x 20 204434147 Silica XG aluminum 200 μm UV254 5 x 20 504434167 Silica XG aluminum 200 μm UV254 5 x 10 5044341C5 Silica XG aluminum 200 μm UV254 5 x 7.5 204434187 Silica XG aluminum 200 μm UV254 4 x 8 504434026 Silica XG aluminum 200 μm - 20 x 20 254434047 Silica XG aluminum 200 μm - 5 x 20 504434067 Silica XG aluminum 200 μm - 5 x 10 5044340C5 Silica XG aluminum 200 μm - 5 x 7.5 20

Hard Layers - Silica HL TLCSilica Hard Layer (HL) is a family of pre-coated glass plates with unmodified silica. It is available in two types as Silica Hard Layer for TLC and Silica Hard Layer for HPTLC. Silica Hard Layer plates differ from the well-known Silica Soft Layer TLC and HPTLC by a special binder system, resulting in the formation of hard, water-resistant and wettable layers with outstanding separation properties and consistent reproducibility from lot to lot.

Like our proven Silica Soft Layer TLC plates, HL TLC Plates are coated with unmodified silica 60. They are available with or without fluorescent indicator (green fluorescence, 254 nm).

If one compares the performance of the HL TLC plates with the conventional Silica Soft Layer TLC plates for the separation of anthraquinone dyes, one observes a similar selectivity, but with different Rf values. A closer observation reveals the slight differences in selectivity for the different layers. For a normal phase separation of anthraquinone dyes, HL TLC and Silica Soft Layer TLC plates behave quite similarly. Under reversed phase conditions, the different characteristics of the two plates become obvious. While the Silica Soft Layer TLC Plate shows some reversed phase properties due to the still lower polarity of the binder system, HL TLC is a true normal phase TLC plate.

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Cat. # Layer Backing Thickness Indicator Size (cm) Qty/pkg1624126 Silica G polyester 200 μm UV254 20 x 20 251624147 Silica G polyester 200 μm UV254 5 x 20 501624187 Silica G polyester 200 μm UV254 4 x 8 501624199 Silica G polyester 200 μm UV254 2.5 x 7.5 2001634126 Silica G aluminum 200 μm UV254 20 x 20 251634147 Silica G aluminum 200 μm UV254 5 x 20 501634187 Silica G aluminum 200 μm UV254 4 x 8 501634199 Silica G aluminum 200 μm UV254 2.5 x 7.5 2001615026 Silica G glass 250 μm - 20 x 20 251615126 Silica G glass 250 μm UV254 20 x 20 251615148 Silica G glass 250 μm UV254 5 x 20 1001615167 Silica G glass 250 μm UV254 5 x 10 501615190 Silica G glass 250 μm UV254 2.5 x 7.5 5001616125 Silica G-Prep glass 500 μm UV254 20 x 20 201617124 Silica G-Prep glass 1000 μm UV254 20 x 20 151618123 Silica G-Prep glass 2000 μm UV254 20 x 20 12

Soft Layers - Silica G TLC• Silica Gel, 60 Å, 5 - 17 μm• Analytical and Preparative Layers (slightly larger particles are used)• Indicator: UV254 - manganese activated zinc silicate with green fluorescence• Polymeric binder which is stable in almost all organic solvents and resistant towards aggressive visualization reagents• Binder for polyester backed plates is completely stable in purely aqueous eluents.


Analysis of Edible Fats Plate: Silica G, w/UV254, polyester backed, 200 μm, 20 x 20 cm, 25/pkPhase: Silica GCatalog #: 1624126Substances: cholesterol triglycerides cholesterol esterInformation: be used without further treatment.Conditions:Eluents: Dichloromethane-toluene 5:2 (v/v) Sample Volume: 0.5 μl Migration Distance: 6 cmDetection: Spray with a ethanolic solution of molybdatophosphoric acid, then dry with hot air. Heat the sheet with a hot air gun for 3-4 min.

Separation of Anthraquinone Dyes Plate: Silica G, w/UV254, glass backed 250 μm, 20 x 20 cm, 25/pk Catalog #: 1615126Eluent: a) Normal Phase TLC: toluene/cylcohexane (2:1. v/v) b) Reversed Phase TLC: acetone/water (1:1. v/v)Migration Distance: a) 4.8 cm in 8 min b) 3.8 cm in 10 minDetection: TLC scanner, UV 254 nmPeaks: 1. Blue 2. Violet 2 3. Blue 1 4. Secondary Spot of Blue

Legend:Compounds Rf Cholesterol 0.16Triglycerides 0.54-0.66 Cholesterol esters 0.95

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Cat. # Layer Backing Thickness Indicator Size (cm) Qty/pkg2315126 Silgur (Silica 60 w/

Diatomaceous earth zone)glass 250 μm UV254 20 x 20 25

2315137 Silgur (Silica 60 w/ Diatomaceous earth zone)

glass 250 μm UV254 10 x 20 50


glass 250 μm - 20 x 20 25


glass 250 μm - 10 x 20 50

Soft Layers - Silica Concentrating Zone TLC PlatesThe concentrating zone serves as a “quick application zone”, which allows quantitative evaluation of chromatograms, even if samples have been applied with an unsteady hand. Since careful sample application is time-consuming or requires special equipment, silica plates with concentrating zone are real time-savers. This includes economy in preparation time because of a reduction in evaporation time: larger volumes of dilute solutions can be applied instead of smaller quantities of concentrated solutions. This is often of advantage especially in biochemical investigations.

Cellulose - Cellulose Layers for TLC PlatesCellulose 300Properties: • Fibre length (95 %) 2 – 20 μm, average degree of polymerisation 400 – 500, specific surface acc. to Blaine• 15000 cm2/g• ≤ 20 ppm Fe, 6 ppm Cu, 7 ppm P; CH2Cl2 extract ≤ 0.25 %; residue on ignition at 850° C ≤ 1500 ppmApplication: • Partition chromatography of polar substances such as amino acids, carboxylic acids or carbohydrates

Cat. # Layer Backing Thickness Indicator Size (cm) Qty/pkg0314026 Cellulose 300 glass 0.10 mm - 20 x 20 250314126 Cellulose 300 glass 0.10 mm UV254 20 x 20 250315026 Cellulose 300 glass 0.25 mm - 20 x 20 250315126 Cellulose 300 glass 0.25 mm UV254 20 x 20 250322126 Cellulose 300 polyester 0.25 mm UV254 20 x 20 250332126 Cellulose 300 aluminum 0.25 mm UV254 20 x 20 25

Cellulose 400Properties: • Prepared: by hydrolysis of high purity cellulose with HCL; mean degree of polymerization 40-200Application: • Carboxylic acids, lower acids, lower alcohols, urea and purine derivatives

Cat. # Layer Backing Thickness Indicator Size (cm) Qty/pkg0412126 Cellulose 400 glass 0.10 mm UV254 20 x 20 250422126 Cellulose 400 polyester 0.10 mm UV254 20 x 20 25

Cellulose 300 PEIProperties: • Fibrous cellulose impregnated with polyethyleneimineApplication: • Analysis of nucleic acids, and of mutagenic substances with the 32P post labelling procedure (see application 402260 at www.sorbtech.com)

Cat. # Layer Backing Thickness Indicator Size (cm) Qty/pkg1122026 Cellulose 300 PEI polyester 0.10 mm - 20 x 20 251122126 Cellulose 300 PEI polyester 0.10 mm UV254 20 x 20 25

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C2 Silica TLC• Base material: Silica Gel, 60 Å, particle size 5 - 17 μm, pH stability 2 - 10• Indicator: UV254 - Acid-resistant product with a pale blue fluorescence; UV-absorbing substances appear as dark-blue to black spots on a light-blue background• Silanized silica with dimethyl modification, carbon content 4%• Recommended applications: active plant constituents, steroids

Cat. # Layer Backing Thickness Indicator Size (cm) Qty/pkg2715126 C18-W Silica glass 250 μm UV254 20 x 20 252715137 C18-W Silica glass 250 μm UV254 10 x 20 502715147 C18-W Silica glass 250 μm UV254 5 x 20 502715156 C18-W Silica glass 250 μm UV254 10 x 10 252717124 C18-W Silica-Prep glass 1000 μm UV254 20 x 20 152733126 C18-W Silica aluminum 150 μm UV254 20 x 20 252733147 C18-W Silica aluminum 150 μm UV254 5 x 20 502733156 C18-W Silica aluminum 150 μm UV254 10 x 10 252733167 C18-W Silica aluminum 150 μm UV254 5 x 10 502733187 C18-W Silica aluminum 150 μm UV254 4 x 8 50

Reverse Phase Layers C18 - W Silica TLC• Base material: Silica Gel, 60 Å, mean particle size 9 μm, pH stability 2 - 10• Indicator: UV254 - Acid-resistant product with a pale blue fluorescence; UV-absorbing substances appear as dark-blue to black spots on a light-blue background• Partial octadecyl (C18) modification, wettable with water, carbon content 14%• Normal phase or reversed phase separation modes with eluents from anhydrous solvents to mixtures with high concentrations of water• Recommended applications: aminophenols, barbiturates, preservatives, nucleobases, polycyclic aromatic hydrocarbons, steroids, tetracyclines, plasticizers (phthalates)

Cat. # Layer Backing Thickness Indicator Size (cm) Qty/pkg2815126 C2 Silica glass 250 μm UV254 20 x 20 250314126 C2 Silica glass 250 μm UV254 10 x 10 250315026 C2 Silica aluminum 150 μm UV254 20 x 20 250315126 C2 Silica aluminum 150 μm UV254 4 x 8 50


Separation of Salicyl DerivativesLayer: C18 w/UV254Sample Volume: 250 nlEluent: methanol/1 N acetic acid (25:75, v/v)Migration distance: 9 cm in 30 minDetection: TLC scanner, UV254 nmPeaks: 1. Salicyluric acid (1.0%) 2. Acetylsalicylic acid (1.0%) 3. Salicyluric acid (0.4%)

Separation of Steroids Layers: C2 w/UV254 Eluent: dichloromethane/methanol (98.5:1.5)Detection: TLC scanner, UV254 nmPeaks: 1. Cortisone 2. Corticosterone 3. Testosterone 4. Deoxycorticosterone 5. Progesterone

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Cat. # Layer Backing Thickness Indicator Size (cm) Qty/pkg3114126 Cyano Nano-Silica glass 200 μm UV254 20 x 20 253114136 Cyano Nano-Silica glass 200 μm UV254 10 x 20 253114156 Cyano Nano-Silica glass 200 μm UV254 10 x 10 253133126 Cyano Nano-Silica aluminum 150 μm UV254 20 x 20 253133187 Cyano Nano-Silica aluminum 150 μm UV254 4 x 8 50

Polar Phase LayersCyano Nano-Silica HPTLC• Base material: Silica Gel, 60 Å, particle size 2 - 10 μm, pH stability 2 - 8• Indicator: UV254 - Acid-resistant product with a pale blue fluorescence; UV-absorbing substances appear as dark-blue to black spots on a light-blue background• Cyanopropyl modification, carbon content 5.5%• Normal phase or reversed phase separation modes depending on the polarity of the developing solvent• Recommended applications: steroid hormones, phenols, preservatives

Amino Nano-Silica HPTLC• Base material: Silica Gel, 60 Å, particle size 2 - 10 μm, pH stability 2 - 8• Indicator: UV254 - Acid-resistant product with a pale blue fluorescence; UV-absorbing substances appear as dark-blue to black spots on a light-blue background• Aminopropyl modification, carbon content 3.5%• Layer can be wetted equally well by pure water as by organic solvents• Recommended applications: vitamins, sugars, steroids, purine derivatives, xanthines, phenols, nucleotides and pesticides

Cat. # Layer Backing Thickness Indicator Size (cm) Qty/pkg3014136 Amino Nano-Silica glass 200 μm UV254 10 x 20 253014156 Amino Nano-Silica glass 200 μm UV254 10 x 10 253033187 Amino Nano-Silica aluminum 200 μm UV254 4 x 8 50

Separation of preservativesLayer: Cyano w/UV254Sample Volume: 400 nlEluent: ethanol/water/glacial acetic acid 20:80:0.2 with 0.1 mol/l tetraethylammonium chlorideMigration distance: 7.3 cm in 30 minDetection: TLC scanner, UV254 nmPeaks: 1. Propyl p-hydroxybenzoate 2. Ethyl p-hydroxybenzoate 3. Methyl p-hydroxybenzoate 4. Benzoic acid 5. Sorbic acid

Separation of preservativesLayer: Amino w/UV254Sample Volume: 500 nlEluent: ethyl acetate/pyridine/water/glacial acetic acid (60:30:10:5, v/v/v/v)Migration distance: 8 cm in 45 min, double developmentDetection: dry layer at 160° C for 5 min, TLC scanner, UV254 nmPeaks (0.1% each): 1. Lactose 2. Saccharose 3. Galactose 4. Glucose 5. Fructose 6. Arabinose 7. Xylose 8. Ribose

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Diol Nano-Silica HPTLC• Base material: Silica Gel, 60 Å, particle size 2 - 10 μm, pH stability 2 - 8• Indicator: UV254 - Acid-resistant product with a pale blue fluorescence; UV-absorbing substances appear as dark-blue to black spots on a light-blue background• Diol modification, carbon content 5.5%• Layer can be wetted equally well by pure water as by organic solvents• Recommended applications: steroids, pesticides and plant constituents; for more critical separations an alternative to silica, since it is less sensitive to water content in the environment; leads to more reproducible results compared to silica

Alumina TLC• Aluminum oxide• Indicator: UV254 - manganese activated zinc silicate with green fluorescence• Recommended applications: terpenes, alkaloids, steroids, aliphatic and aromatic compounds• We recommend heating the plates for 10 min. at 120° C before usage to activate the alumina layer.

Cat. # Layer Backing Thickness Indicator Size (cm) Qty/pkg2914156 Diol Nano-Silica glass 200 μm UV254 10 x 10 252933187 Diol Nano-Silica aluminum 150 μm UV254 4 x 8 50

Cat. # Layer Backing Thickness Indicator Size (cm) Qty/pkg0115126 Alumina glass 250 μm UV254 20 x 20 250115148 Alumina glass 250 μm UV254 5 x 20 1000117134 Alumina-Prep glass 1000 μm UV254 20 x 20 150224126 Alumina polyester 200 μm UV254 20 x 20 250224147 Alumina polyester 200 μm UV254 5 x 20 500224187 Alumina polyester 200 μm UV254 4 x 8 500224199 Alumina polyester 200 μm UV254 2.5 x 7.5 2000234126 Alumina aluminum 200 μm UV254 20 x 20 250234147 Alumina aluminum 200 μm UV254 5 x 20 50


Separation of pesticidesLayer: Diol w/UVA254Sample Volume: 2 μlEluent: petroleum ether (40 - 60° C)/acetone (80 +20, v/v)Migration distance: 7 cm Detection: TLC scanner, 238 nmPeaks (0.07% each in MeOH): 1. Metoxuron 2. Monuron 3. Metobromuron

Separation of lipophilic dyesLayer: Alumina w/UVA254Sample Volume: 1000 nlEluent: toluene/cyclohexane (2:1,v/v) Migration distance: 10.8 cm in 15 minDetection: TLC scanner, 238 nmPeaks: 1. Indophenol 2. Sudan red G 3. Sudan blue II 4. Butter yellow

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Chiral TLCSorbtech offers TLC Chiral plates: reverse phase silica gel impregnated with a chrial selector and copper ions. This allows for multiple simultaneous separations – an ideal plate for routine applications and production control. The separation of optically active isomers is based on ligand exchange. The chrial selector on the plate and the enantiomers to be separated form diastereomeric mixed chelate complexes with the transition metal ion. Complexes for the different antipodes have different stabilities thus achieving chromatographic separation.

Cat. # Layer Backing Thickness Indicator Size (cm) Qty/pkg3415126 Chiral glass 250 μm UV254 20 x 20 253415136 Chiral glass 250 μm UV254 10 x 20 253415131 Chiral glass 250 μm UV254 10 x 20 43415147 Chiral glass 250 μm UV254 5 x 20 503415156 Chiral glass 250 μm UV254 10 x 10 25

Quantitative determination (remission location curves) of TLC-separated enantiomers of tert.-leucine:

Layer: CHIRAL TLC Eluent: methanol – water (10:80, v/v) Detection: dip in 0.3% ninhydrin solution quantification with scanner, 520 nm

a) L-tert.-leucine,b) L-tert.-leucine + 0.1% D-tert.-leucine, c) L-tert.-leucine + 1 % D-tert.-leucine, d) external reference sample

TLC Accessories

Every lab needs a TLC Glass Plate Cutter.

Save Time • Just score and snap off • High quality carbide scriber is mounted into a movable plastic head for optimal, one-finger scoring• Virtually maintenance-free, a standard screwdriver disassembles cutter for easy cleaning and care

Save Money• No need to share experiments in the classroom• Use only what you need, small runs are now cost effective• Benefit from bulk purchases then select your size

Cat.# TLC-Cutter

Cat.# 814019 Cat.# 814022 Cat.# 814023

Developing Chamber Glass Capillaries

TLC Spotting Guide

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Thin Layer Chromatography - Sorbent Technologies · PDF fileThin Layer Chromatography Sorbent...

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Transcript of Thin Layer Chromatography - Sorbent Technologies · PDF fileThin Layer Chromatography Sorbent...