TLC PDF

Thin layer chromatography
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Basics
Thin layer chromatography (TLC) and high performance thin the user to carry out some additional steps for preparing a sam-
layer chromatography (HPTLC), also called planar chromatogra- ple, thus introducing the user to techniques often performed in
phy, are, like all chromatographic techniques, based on a multi- industrial laboratories.
stage distribution process involving Thorough preparation of samples is an important prerequisite for
· Suitable adsorbents (the stationary phase) coated as a thin the success of a TLC separation. For our range of products for
layer onto a suitable support (e.g., glass plate, polyester or more demanding sample pretreatment please see the chapter
aluminum sheet; also see page 272) “SPE” from page 10.
· Solvents or solvent mixtures (the mobile phase or eluent) Sample application
· Sample molecules The most frequent technique is application with a glass capillary
The principle of TLC is known for more than 100 years [11]. as spot or short streak.
The real break-through as an analytical method, however, came Application as streak will yield better results especially for instru-
about 50 years ago as a consequence of the pioneering work mental quantification. For both types of application some man-
of Egon Stahl [12]. ual skill is required to obtain reproducible results. Substance
Today TLC has gained increasing importance as an analytical zones which are too large from the beginning will cause poor
separation technique, which is probably due to effects of instru- separation since during chromatography they will become even
mentation and automation [13]. At the same time the applicabil- larger and more diffuse.
ity of thin layer chromatography was enhanced by development A valuable aid for manual application especially of large vol-
of new adsorbents and supports. umes of very dilute samples is the concentrating zone (e.g.,
Today MACHEREY-NAGEL offers a versatile range of ready-to- SILGUR-25 UV254), which consists of a chromatographically in-
use layers, which are the result of 50 years of continuous re- active adsorbent (kieselguhr). The substances to be separated
search and development. are concentrated to a small band in the concentrating zone and
the separation starts at the beginning of the chromatographically
Features of modern TLC / HPTLC active adsorbent silica.
The success of thin layer chromatography as a highly efficient

microanalytical separation method is based on a large number
of advantageous properties:
· High sample throughput in a short time Silica layer
· 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 ready-to-use layer acts as
Concentrating zone
storage medium for data)
· Separated substances can be subjected to subsequent ana- Arbitrary application Developed
lytical procedures (e.g., IR, MS) at a later date in the concentrating chromatogram
zone
· Rapid and cost-efficient optimization of the separation due to
easy change of mobile and stationary phase Another method for sample concentration is a short pre-elution
(few mm) with a solvent, in which all substances have a high Rf
Principle steps of a TLC separation value.
If a quantitative evaluation with a TLC scanner is to follow the
Sample preparation separation we recommend to use commercially available sam-
For separation the sample must meet several requirements to ple applicators for spotting. These range from simple spotting
obtain good results. Since the TLC plate is a disposable prod- guides via nanoapplicators to completely automated spotting
uct, sample preparation in general is not as demanding as for devices. Application as streak can be performed automatically
other chromatographic methods. However, eventually several by spraying of the sample without touching the layer of the TLC
steps for sample pretreatment may be necessary. These include plate. Application as band over the whole width of the TLC plate
sampling, mechanical crushing, extraction steps, filtration and is especially important for preparative TLC. After application al-
sometimes enrichment of interesting components or clean-up, low the solvent of the samples to evaporate completely (about
i.e. removal of undesired impurities. 10 min) or blow with cold or hot air. Development of a chro-
Our TLC micro-sets introduce some simple methods of sample matogram should never start before the solvent of the applied
pretreatment. The dyes or dye mixtures of the beginner’s set do samples is evaporated completely.
not require complicated procedures. The advanced sets require
266 www.mn-net.com
Basics
Developing a chromatogram – separation techniques Quantitative evaluation is possible by suitable calibration mea-
The most frequently used separation technique is ascending surements. For this purpose either the area of a substance spot
TLC in a trough chamber (standard method, linear develop- is measured or a photometric evaluation is performed directly
ment). Usually it is applied as single development. However, on the layer. The latter procedure, however, requires a higher
multiple development, with or without change of eluent (step instrumental expense.
technique) can improve separation results. For 2-dimensional The following paragraphs describe the most frequently used
development only 1 spot of the sample is applied in one edge methods for evaluation in TLC.
of a plate. After chromatography in the first direction the plate is
dried, turned by 90° and developed in the 2nd dimension with Solvent front
another eluent. Thus complicated mixtures give 2-dimension-
al chromatograms taking advantage of the different separating
properties of two eluents.
Substance spot
For selection and optimization of the eluent numerous publica-
a
tions are available. A generally applicable standardized optimiza-
tion method is described by H. Keuker et al. [14]. b
It is important to pay attention to the atmosphere in the de-

veloping chamber. If reproducible migration distances are re-
quired, saturation of the chamber atmosphere with eluent vapor Starting line
is necessary. For this purpose the developing chamber is lined
with well absorbing chromatography paper (e.g., MN 260) and
Qualitative detection
charged with a correspondingly larger volume of eluent.
Qualitative evaluation is generally made directly on the TLC plate
via characteristic Rf values of substances, i.e. the ratio of di-
stance start – substance zone to distance start – solvent front
and specific chemical reactions.
Support with
adsorbent layer Visualization of separated substances
First of all it is necessary to recognize the position of a sub-
stance spot. Only in very few cases the sample is a dye which
Solvent vapor
can be seen with the naked eye. Much more often for unspecific
visualization substances can be viewed under UV light, since
many substances show a UV absorption. If a fluorescent indi-
Filter paper
cator is added to the layer, all substances absorbing in the re-
spective region of wave length cause a quenching of the fluores-
cence, i.e. they appear as dark spots on the fluorescent layer.
Eluent
Customary fluorescent indicators are excited at 254 nm or (less
A) Normal saturation, arrows
show evaporation of the
B) Chamber lined with filter
paper, saturated with frequently) at 366 nm with a mercury lamp. For our program of
eluent from the layer eluent vapor fluorescent indicators for TLC please see page 296.
Evaluation of a thin layer chromatogram

Evaluation depends on the purpose of the chromatographic
analysis. For qualitative determination often localization of sub-
stances is sufficient. This can be easily achieved by parallel runs
with reference substances.
A parameter often used for qualitative evaluation is the Rf value
(retention factor) or the 100-fold value hRf. The Rf value is defi-
ned as follows: Quenching of the fluorescence
Identification of separated substances is possible via the Rf val-

distance starting line – middle of spot b
Rf = = ue compared to the pure compound, which is often applied si-
distance starting line – solvent front a multaneously on the same plate.
i.e. the Rf values are between 0 and 1, best between 0.1 and 0.8 For a number of compounds their native fluorescence can be
(i.e. 10–80 for hRf). If reproducible Rf values are to be obtained, it used for visualization, which is excited by UV light (mostly long-
is essential that several parameters such as chamber saturation, wave UV) (e.g., aflatoxins). This allows not only determination of
composition of solvent mixtures, temperature etc. are strictly the Rf value, but often enables a further qualitative assignment.
controlled.
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Basics
If these methods do not allow localization or characterization evaluation of circular or anti-circular chromatograms with very
of a substance, post-chromatographic detection methods can high ease of operation. In addition to manual operation control
be applied, chemical reactions on the TLC plate [15]. Quite un- by a computer is possible with respective data collection and
specific reactions are iodine adsorption and the charring tech- storage. Usually wavelengths from 200 to 700 nm are available
nique (spraying with sulfuric acid and heat treatment). (visible and UV), e.g., all post-chromatographic (and of course
More reliable results are possible with specific reagents for all pre-chromatographic) visualization procedures are evaluated
spraying or dipping, which form colored or fluorescent com- with the proper wavelength, which is determined with the instru-
pounds with the substances to be detected. Depending on the ment. Time requirements for all these possibilities are extremely
sensitivity of these reactions they are not only used for group or low. Interlaboratory experiments with standard deviations of 2 %
substance specific characterization (in addition to the Rf value) show how excellent results are obtainable [16].
but also for quantification down to trace levels. As example take
the ninhydrin reaction. Formation of a (usually red) zone with this
detection method yields the information, that a certain group
of substances, e.g., α-amino acids, are present. The Rf value
allows further assignment to one or several single compounds.
For identification of a substance a combination of different de-
tection methods can be useful. Thus almost all lipids can be
converted to products with light green fluorescence by reaction
with 2’,7’-dichlorofluorescein. Adsorption of iodine vapor en-
ables a differentiation between saturated and unsaturated lipids
or lipids containing nitrogen. And finally the Rf value is a third
means of identification.
Here are some general remarks concerning spraying: use all
spray reagents under a fume hood. The developed, dried TLC
plate or sheet is placed on a sheet of filter paper for spraying.
Usually it is sufficient to fill the sprayer with about 5–10 mL solu-
tion. Spray from a distance of about 15 cm with the aid of a rub-
ber ball or – if available – with pressurized air. It is always better
to spray a layer twice very thinly and evenly (with intermediate
drying), than to saturate the layer with excessive spray reagent.
In the latter case spots tend to become diffuse. After visualiza-
tion mark outlines of zones with a lead pencil, because some
spots tend to fade after a while.
Especially for quantitative evaluation short dipping of the layer in
the respective reagent solution is recommended. For this pur-
pose automatic instruments are commercially available, which
allow reproducible dipping.
When a substance is localized on the TLC plate (e.g., under
UV), but not yet identified, TLC scanners allow recording of UV
spectra of individual substance zones directly on the layer, or
the zone is removed by scratching or cutting (for sheets), elut-
ed and further analyzed, e.g., by FT-IR, RAMAN, NMR or mass
spectroscopy.
Quantitative evaluation
Often TLC is considered to be only a semiquantitative analytical
procedure. This is true for visual evaluation of spots, since the eye
can only compare but not measure absolute values. If, however,
a direct optical evaluation (“in situ” measurement) is performed
on the TLC plate with a thin layer scanner, after measurement
of calibration functions, exact quantitative results are possible.
Commercial scanners offer many features such as evaluation
in absorption and fluorescence, unattended programmed scan-
ning of lanes, multi-wave length measurement, background cor-
rection, selectable base line for integration, recording of spectra,
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Introductory kits
TLC micro-sets introductory kits for science education

Beginner’s set Advanced sets F1, F2 and F3
· Features separations with simple developing solvents; sam- · Require some experience and skill from the user: some of
ples are colored thus eliminating the need for visualization. the samples have to be pretreated before separation, and
· All equipment needed is contained in the set. for identification of substances spray reagents have to be
used
TLC micro-set A for beginners Contents of TLC micro-set A for beginners

This kit contains all chemicals and accessories for the following 1 manual
separations: 3 developing chambers
· Separation of the fat-soluble (lipophilic) 50 glass capillaries 1 μL
Test dye mixture 1: butter yellow, indophenol, sudan blue II,
1 spotting guide
sudan red G
2 felt tip pens
· Separation of a mixture of anthraquinone dyes
Test dye mixture 2: blue 1, blue 3, green, green blue, red, 1 measuring cylinder 10 mL
violet 1, violet 2 50 polyester sheets 4 x 8 cm each of POLYGRAM®:
· Separation of a mixture of food dyes SIL G/UV254, Alox N/UV254 and CEL 300
Test dye mixture 3: brilliant black BN (E151), fast red E, 8 mL each of test dye mixture 1 (4 lipophilic dyes),
erythrosine (E127), yellow orange S (sunset yellow CFC, test dyes sudan red G, and sudan blue II
E110), naphthol red S, ponceau 4 R (E124), tartrazine (E102) 8 mL each of test dye mixture 2 (7 anthraquinone dyes),
· Separation of dyes from felt tip pens test dyes blue 1 and violet 2
8 mL each of test dye mixture 3 (7 food dyes),
test dyes yellow orange S, and brilliant black BN
100 mL each of toluene, toluene – cyclohexane (2:1, v/v),
ethanol, 2.5 % sodium citrate solution,
25 % ammonia solution – 2-propanol (5:3, v/v)
Ordering information
Designation Pack of REF
TLC micro-set A for beginners* 1 kit 814000
Replacement parts for TLC micro-set A
Test dye mixture 1*, solution of 4 lipophilic dyes in toluene (components see above) 8 mL 814001
Test dye mixture 2*, solution of 7 anthraquinone dyes in toluene – cyclohexane (2:1, v/v) (components see above) 8 mL 814002
Test dye mixture 3, aqueous solution of 7 food dyes (components see above) 8 mL 814003
Collection of 4 individual components of test dye mixture 1* 4 x 8 mL 814011
Collection of 7 individual components of test dye mixture 2* 7 x 8 mL 814012
Collection of 7 individual components of test dye mixture 3 7 x 8 mL 814013
Sodium citrate, 2.5 g in 100 mL bottle to fill up with distilled water 2.5 g 814029
* These products contain harmful substances which must be specially labeled as hazardous. For detailed information please see SDS.
Information about the advanced sets F1, F2 and F3 can be found on page 270 and page 271.
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Introductory kits
TLC micro-set F1 Contents of TLC micro-set F1
This kit contains all chemicals required for the separation of 1 manual, 50 glass capillaries 1 μL
· Amino acids (test mixture, consisting of alanine, arginine, 50 polyester sheets 4 x 8 cm each of POLYGRAM®:
tryptophan and valine) SIL G/UV254 and CEL 300
· Amino acids in urine 100 mL each of n-butanol, ninhydrin spray reagent (0.2 %
· The heavy metal cations copper(II) and manganese(II) in ethanol), acetone, 25 % ammonia solution, rubeanic acid
spray reagent
50 mL each of 50 % acetic acid, 18 % hydrochloric acid
8 mL each of the amino acid test mixture (see left), tryptophan
and arginine reference solutions
8 mL each of the heavy metal cation test mixture (see left),
Cu2+ and Mn2+ reference solutions

This kit contains all chemicals required 1 manual, 50 glass capillaries 1 μL
· For analysis of edible fats 50 polyester sheets 4 x 8 cm POLYGRAM®:
· For analysis of fats and cholesterol in blood SIL G/UV254
5 disposable pipettes 25 μL
5 sample vials N 11 (1.5 mL) with PE caps and seals
3 sample vials 30 mL (for butter, margarine and edible oil)
100 mL each of cyclohexane and molybdatophosphoric acid
spray reagent
2 x 50 mL acetone with calibrated pipette
25 mL butan-2-one
8 mL cholesterol reference solution

This kit contains all chemicals required 1 manual, 50 glass capillaries 1 μL
· For separation of analgetics (pain relievers) 50 polyester sheets 4 x 8 cm POLYGRAM®:
· For drug analysis as shown for cinchona bark SIL G/UV254
5 Aspirin® tablets, 5 Thomapyrin® tablets
20 folded filters MN 615 1/4, 11 cm diameter
3 sample vials 8 mL (for Aspirin® sample, Thomapyrin® sam-
ple, cinchona bark extract), 5 g cinchona bark
100 mL each of ethanol, 2-propanol, toluene – diethyl ether
je 100 mL Ethanol, 2-Propanol, Toluol – Diethylether (61:39,
v/v), spray reagent for caffeine and spray reagent according
to Dragendorff-Munier
50 mL each of iron(III) chloride solution and potassium hexa-
cyanoferrate(III) solution, 30 mL ethyl acetate
25 mL each of 12.5 % ammonia solution and diethylamine
8 mL each of caffeine, paracetamol, quinine reference solu-
tions
All experiments with TLC micro-sets F1–F3 require the materials kit (see TLC micro-set M on page 271).
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Introductory kits
TLC micro-set F1* 1 kit 814200
Refill reagents for TLC micro-set F1
Amino acid test mixtures (components see previous page) 8 mL 814201
Collection of 4 individual components of the amino acid test mixture 4 x 8 mL 814202
Cation test mixture (components see previous page) 8 mL 814204
Collection of 2 individual components of the cation test mixture (Cu2+, Mn2+) 2 x 8 mL 814205
Cholesterol reference solution* 8 mL 814301
Quinine reference solution* 8 mL 814405
Paracetamol reference solution* 8 mL 814406
Caffeine reference solution* 8 mL 814407
Refill packs TLC sheets for all TLC micro-sets
TLC polyester sheets POLYGRAM® SIL G/UV254, 4 x 8 cm 4 x 50 814025
TLC polyester sheets POLYGRAM® Alox N/UV254, 4 x 8 cm 4 x 50 814026
TLC polyester sheets POLYGRAM® CEL 300, 4 x 8 cm 4 x 50 814027
TLC polyester sheets POLYGRAM® 4 x 8 cm: 100 x SIL G/UV254; 50 x Alox N/UV254; 50 x CEL 300 1 kit 814028
* These products contain harmful substances which must be specially labeled as hazardous. For detailed information please see SDS.
Accessories for TLC micro-sets can be found under TLC accessories on page 295.
Spray reagents can be found on page 296.
TLC micro-set M Contents of TLC micro-set M (materials kit)

This kit is prerequisite for the separations with kits F1 to F3. In 2 x 50 glass capillaries 1 μL, 2 spotting guides
addition, it serves as basic equipment for the individual study of 1 rubber cap for capillaries
further thin layer chromatographic experiments.
1 measuring cylinder 10 mL
1 beaker 25 mL
2 developing chambers
1 glass laboratory sprayer with rubber bulb
1 plastic syringe 1 mL
20 sheets filter paper MN 713 (15 x 21 cm)
50 polyester sheets 4 x 8 cm each of POLYGRAM®:
SIL G/UV254, Alox N/UV254 and CEL 300
TLC micro-set M (materials kit) 1 kit 814100
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Summary of MN ready-to-use layers
Advantages of MN plates and sheets for TLC Adsorbents for MN plates and sheets for TLC
Continuous high quality Classical adsorbents
· Guaranteed by stringent production control including stan- · For ~ 80 % of all TLC separations silica 60 (mean pore diam-
dardized lot tests, surface checks for roughness or cracks as eter 60 Å = 6 nm) is used
well as hardness and adherence checks · Other classical adsorbents are aluminum oxide, cellulose,
Comprehensive range of phases for TLC / HPTLC kieselguhr, ion exchangers and polyamide
· There is no universal TLC plate which meets all possible Special phases
types of analyses · Modified silica, like C18 (octadecyl-) cyano-, amino-, diol-,
· Our versatile range of TLC ready-to-use layers covers many RP-2
different types of applications · Special layers for specific separations, like PAH- or enantio-
Immediately ready for chromatographic separation mer separation
· Coatings or impregnations are not necessary Particle size distribution and thickness of layer
Homogeneous, smooth, well adhering layers · Are chosen to fit the given type of application (e.g., HPTLC,
standard or preparative separations)
· An important criterion especially for reproducible quantitative
evaluation · Most MN ready-to-use layers are available with or without
fluorescent indicator
Electron microscope photograph of a cross section through a glass plate Electron microscope photograph of a cross section through an aluminum
with silica layer (magnification x 500) sheet with silica layer (magnification x 500)
Supports for ready-to-use layers for TLC
Glass plates G POLYGRAM® P ALUGRAM® A / ALUGRAM® Xtra Ax

Physical properties of support materials
Material glass polyester aluminum
Thickness (approx.) 1.3 mm 0.2 mm 0.15 mm
Weight, packaging and storage requirements high low low
Torsional strength ideal low relatively high
Temperature stability high max. 185 °C high
Susceptible to breakage yes no no
Can be cut with scissors no yes yes
Chemical resistance of support materials
Against solvents high high high
Against mineral acids and conc. ammonia high high low
Stability of the binder system of NP plates in water
Suitability for aqueous detection reagents depending on very suitable ALUGRAM®: limited suitability;
phase ALUGRAM® Xtra: very suitable
272 www.mn-net.com
Summary of MN ready-to-use layers
Summary
Phase Support* Layer Page
Standard silica particle size 5–17 µm
ADAMANT G silica 60, improved binder system, optimized particle size distribution 274
SIL G G P A Ax silica 60, standard grade 276
DURASIL G silica 60, special binder system 277
SILGUR G Ax silica 60 with kieselguhr concentrating zone 279
Unmodified silica for HPTLC particle size 2–10 µm

Nano-SILGUR G Ax nano silica 60 with kieselguhr concentrating zone 279
Nano-ADAMANT G nano silica 60, improved binder system, optimized particle size distribution 281
Nano-SIL G A Ax nano silica 60, standard grade 281
Nano-DURASIL G nano silica 60, special binder system 282
Modified silica for HPTLC particle size 2–10 µm

Nano-SIL C18-50 /
G nano silica with partial or complete C18 modification 283
Nano-SIL C18-100
RP-18 W/UV254 G A nano silica with partial octadecyl modification, wettable with water 284
RP-2/UV254 G A silanized silica = dimethyl-modified nano silica 60 284
Nano-SIL CN G A cyano-modified nano silica 285
Nano-SIL NH2 G A amino-modified nano silica 286
Nano-SIL DIOL G diol-modified nano silica 287
Aluminum oxide
Alox-25 / Alox N G P A aluminum oxide 288
Cellulose, unmodified and modified

CEL 300 G P A native fibrous cellulose MN 300 289
CEL 400 G P microcrystalline cellulose MN 400 (AVICEL®) 289
CEL 300 PEI P polyethyleneimine-impregnated cellulose ion exchanger 290
CEL 300 AC P acetylated cellulose MN 300 290
POLYAMID-6
POLYAMID-6 P perlon = ε-polycaprolactame 290
Layers for special separations

CHIRALPLATE G RP silica with Cu2+ ions and chiral reagent, for enantiomer separation of amino acids 291
SIL N-HR P high purity silica 60, special binder system, higher gypsum content 291
SIL G-25 HR G high purity silica 60 with gypsum, recommended for aflatoxin analysis 292
SIL G-25 Tenside G silica G with ammonium sulfate for separation of surfactants 292
Nano-SIL PAH G nano silica with special impregnation for PAH analysis 292
IONEX-25 SA-Na P mixed layer of strongly acidic cation exchanger and silica 293
IONEX-25 SB-AC P mixed layer of strongly basic anion exchanger and silica 293
Alox / CEL-AC-Mix G mixed layer of aluminum oxide and acetylated cellulose 293
SILCEL-Mix G mixed layer of cellulose and silica 293
* G = Glass plates P = POLYGRAM® polyester sheets A = ALUGRAM® aluminum sheets Ax = ALUGRAM® Xtra aluminum sheets
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Unmodified TLC silica layers
ADAMANT G unmodified standard silica layers
Key features Technical characteristics

· Outstanding hardness and abrasion resistance due to an · Silica 60, mean pore size 60 Å, specific surface (BET)
optimized binder system ~ 500 m2/g, specific pore volume 0.75 mL/g, particle size
· Increased separation efficiency due to an optimized particle 5–17 μm
size distribution
· High suitability for trace analysis resulting from a UV indicator
with increased brilliance and a lownoise background of the
layer
Separation of steroids Separation of barbiturates

MN Appl. No. 402930 MN Appl. No. 402950
Layers: ADAMANT UV254, SIL G/UV254 Layer: ADAMANT UV254
Sample: 0.1 % solution in CHCl3 Sample volume: 1 μL
Eluent: chloroform – methanol (97:3, v/v) Eluent: chloroform – acetone (95:5, v/v)
Migration distance: ADAMANT 50 mm in 10 min, SIL G 57 mm in 10 min Migration distance: 70 mm in 20 min
Detection: UV Detection: UV
ADAMANT UV254
ADAMANT UV254 SIL G/UV254
Substance Rf ADAMANT Rf SIL G Substance Rf

Cortisone 0.37 0.27 Thiamylal (0.5 %) 0.69
Corticosterone 0.43 0.30 Thiopental (1.0 %) 0.65
Testosterone 0.50 0.39 Hexobarbital (5.0 %) 0.41
Deoxycorticosterone 0.55 0.46 Pentobarbital (1.0 %) 0.26
Progesterone 0.73 0.62 Phenobarbital (1.0 %) 0.18
Plate size [cm] 2.5 x 7.5 5 x 10 5 x 10 5 x 20 10 x 10 10 x 20 20 x 20 Thickness of layer Fluorescent indicator
Pack of [plates] 100 50 200 100 25 50 25
Glass plates
ADAMANT 821040 821040.200 821050 821060 0.25 mm –
ADAMANT UV254 821005 821010 821010.200 821015 821020 821025 821030 0.25 mm UV254
274 www.mn-net.com
ALUGRAM® Xtra SIL G Ax unmodified standard silica layers on aluminum

· Outstanding wettability for precise colorization results, even · Silica 60, mean pore size 60 Å, specific surface (BET)
with 100 % aqueous detection reagents ~ 500 m2/g, specific pore volume 0.75 mL/g, particle size
· Excellent separation efficiency and reproducibility from lot to 5–17 μm
lot · Binder: highly polymeric product, which is stable in almost
· Easy and reliable cutting due to an optimized binder system, all organic solvents and resistant towards aggressive visu-
no flaking of silica alization reagents, also completely stable in purely aqueous
eluents
Separation of nutmeg ingredients

MN Appl. No. 403590
Anethole
Layer: ALUGRAM® Xtra SIL G UV254 Myristicin
Sample: shake 1.0 g freshly powdered drug for 3 min with
4 mL methanol and filter;
apply 10 μL
Eluent: toluene – ethyl acetate (95:5, v/v) Eugenol
Migration distance: 15 cm
Detection: 254 nm: underivatized Linalool
daylight and 366 nm: spray with 5 % ethanolic
sulfuric acid, 1 % vanillic acid and heat to 105 °C
The chromatograms show the following zones with increasing Rf values:

linalool (bluish grey), eugenol (yellowish brown), myristicin (reddish
brown), and anethole (pink-violet). Other colored zones may appear. Daylight 254 nm 366 nm
Plate size [cm] 2.5 x 7.5 4x8 5 x 7.5 5 x 10 5 x 20 10 x 20 20 x 20 Thickness of layer Fluorescent indicator
Pack of [plates] 200 50 20 50 50 20 25
ALUGRAM® Xtra aluminum sheets
SIL G 818230.20 818261 818232 818233 0.20 mm –
SIL G/UV254 818329 818331 818330.20 818360 818332 818362 818333 0.20 mm UV254
Further application examples can be found online in our application database at www.mn-net.com/apps
www.mn-net.com 275
SIL G G P A unmodified standard silica layers
Technical characteristics
· Silica 60, mean pore size 60 Å, specific surface (BET) · Indicators: manganese activated zinc silicate with green
~ 500 m2/g, specific pore volume 0.75 mL/g, particle size fluorescence for short-wave UV (254 nm); special inorganic
5–17 μm fluorescent pigment with blue fluorescence for long-wave
· Thickness of layer for analytical plates 0.25 mm, for prepar- UV (366 nm)
ative plates 0.5 and 1 mm; for 2 mm preparative layers a · Binders: highly polymeric products, which are stable in
slightly coarser material is used almost all organic solvents and resistant towards aggres-
sive visualization reagents; binder system for POLYGRAM®
sheets is also completely stable in purely aqueous eluents
Glass plates
Plate size [cm] 2.5 x 7.5 5 x 10 5 x 10 5 x 20 10 x 10 10 x 20 20 x 20 Thickness of layer
Pack of [plates] 100 50 200 100 25 50 25
SIL G-25 809017 809017.200 809011 809012 809013 0.25 mm
SIL G-25 UV254 809028.100 809027 809027.200 809021 809020 809022 809023 0.25 mm
SIL G-25 UV254+366 809121 809122 809123 0.25 mm
Glass plates
Pack of [plates] (preparative TLC) 20
SIL G-50 809051 0.50 mm
SIL G-50 UV254 809053 0.50 mm
Glass plates
SIL G-100 809061 1.00 mm
SIL G-100 UV254 809063 1.00 mm
Glass plates
SIL G-200 809073 2.00 mm
SIL G-200 UV254 809083 2.00 mm
POLYGRAM® polyester sheets
Plate size [cm] 2.5 x 7.5 4x8 5 x 20 20 x 20 40 x 20
Pack of [plates] 200 50 50 25 25
SIL G 805902 805032 805012 805013 805014 0.20 mm
SIL G/UV254 805901 805021 805022 805023 805024 0.20 mm
SIL G/UV254 roll 500 x 20 cm 805017 0.20 mm
ALUGRAM® aluminum sheets
Plate size [cm] 2.5 x 7.5 4x8 5 x 7.5 5 x 10 5 x 20 10 x 20 20 x 20
Pack of [plates] 200 50 20 50 50 20 25
SIL G 818030.20 818161 818032 818163 818033 0.20 mm
SIL G/UV254 818129 818131 818130.20 818160 818132 818162 818133 0.20 mm
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DURASIL G unmodified standard silica layers
· Silica 60, mean pore size 60 Å, specific surface (BET) · Hard, water-resistant and wettable layers due to a special
~ 500 m2/g, specific pore volume 0.75 mL/g, particle size binder system
5–17 μm
Plate size [cm] 5 x 10 5 x 10 5 x 20 10 x 20 20 x 20 Thickness of layer Fluorescent indicator
Pack of [plates] 50 200 100 50 25
Glass plates
DURASIL-25 812003 812004 0.25 mm –
DURASIL-25 UV254 812005 812005.200 812006 812007 812008 0.25 mm UV254
The most TLC layers are available as glass plate, polyester- or aluminum sheet (also see page 272 and 273).
www.mn-net.com 277
Silica layers with concentrating zone
MN TLC pre-coated layers

– qualitative and individual tailored
Kieselguhr zone
· For rapid sample application
· Because kieselguhr is completely inert towards a large number of compounds, the samples always form a narrow band
at the interface of the two adsorbents, irrespective of shape, size or position of the spots in the concentrating zone.
Separation then takes place in the silica layer.
Silica layer
Concentrating zone
Arbitrary application Applied substances Applied substances Developed

in the concentrating are concentrated at the phase chromatogram
zone boundary
278 www.mn-net.com
Silica layers with concentrating zone
SILGUR G Ax unmodified standard silica layers with concentrating zone
· Silica 60, mean pore size 60 Å, specific surface (BET) · Channel-plate with 19 channels help to prevent cross con-
~ 500 m2/g, specific pore volume 0.75 mL/g, particle size tamination by separating several samples
5–17 μm · More samples can be separated on a plate, and spot areas
· Kieselguhr zone for rapid sample application (see page can be more easily determined
278)
Plate size [cm] 10 x 20 20 x 20 Thickness of layer Fluorescent indicator
Glass plates
Pack of [plates] 50 25
SILGUR-25 810012 810013 0.25 mm –
SILGUR-25 UV254 810022 810023 0.25 mm UV254
Channel-Plates
Pack of [plates] 25
SILGUR-25-C UV254 810123 0.25 mm UV254
ALUGRAM Xtra aluminum sheets
®
SILGUR 818412 818413 0.20 mm –
SILGUR UV254 818422 818423 0.20 mm UV254
Nano-SILGUR G Ax unmodified HPTLC silica layers with concentrating zone
· Nano silica 60, pore size 60 Å, specific surface (BET) · Kieselguhr zone for rapid sample application
~ 500 m2/g, mean specific pore volume 0.75 mL/g, particle (see page 278)
size 2–10 μm
Plate size [cm] 10 x 10 Thickness of layer Fluorescent indicator
Pack of [plates] 25
Glass plates
Nano-SILGUR-20 811032 0.20 mm –
Nano-SILGUR-20 UV254 811042 0.20 mm UV254
Nano-SILGUR 818432 0.20 mm –
Nano-SILGUR UV254 818442 0.20 mm UV254
www.mn-net.com 279
Unmodified HPTLC silica layers
Sharper separation
by nano silica
Nano silica for HPTLC

· Narrow fractionation of the silica particles allows theoreti-
2–10 µm
cal plate heights, which are one order of magnitude smal-
ler than on standard silica layers.
Advantages
· Shorter migration distances 5–17 µm
· Lower amount of samples required Nano silica
· Increased detection sensitivity with equal selectivity
· Less developing time
Standard silica
Comparison of ADAMANT and Nano-ADAMANT plates for separation of anthraquinone dyes

Layers: A) ADAMANT A)
1
B) Nano-ADAMANT
Sample: 1 μL, about 0.1 %
Eluent: toluene – cyclohexane (4:3, v/v)
Migration time: A) 30 min, B) 15 min
2
Peaks:
1. Blue 3 4 5
3 7
2. Violet 2
6
3. Red
4. Green
5. Blue 1 10 20 30 40 50 mm
6. Greenish blue
7. Violet 1
B)
1
5
4
3 7
6
10 20 30 40 mm
280 www.mn-net.com
Nano-ADAMANT G unmodified HPTLC silica layers

· Outstanding hardness and abrasion resistance due to an · Nano silica 60, mean pore size 60 Å, specific surface (BET)
optimized binder system ~ 500 m2/g, specific pore volume 0.75 mL/g, particle size
· Increased separation efficiency due to an optimized particle 2–10 μm
size distribution
· High suitability for trace analyses resulting from a UV indica-
tor with increased brilliance and a lownoise background of
the layer
Glass plates
Nano-ADAMANT 821140 821150 0.20 mm –
Nano-ADAMANT UV254 821110 821120 0.20 mm UV254
Nano-SIL G Ax A unmodified HPTLC silica layers
· Nano silica 60, mean pore size 60 Å, specific surface (BET) · Binder: highly polymeric product, which is stable in almost
~ 500 m2/g, specific pore volume 0.75 mL/g, particle size all organic solvents and resistant towards aggressive visual-
2–10 μm ization reagents
· Indicator: manganese activated zinc silicate with green fluo-
rescence for short-wave UV (254 nm)
Plate size [cm] 5x5 5 x 20 10 x 10 10 x 20 20 x 20 Thickness of layer Fluorescent indicator
Pack of [plates] 100 50 25 50 25
Glass plates
Nano-SIL-20 811011 811012 811013 0.20 mm –
Nano-SIL-20 UV254 811021 811022 811023 0.20 mm UV254
Nano-SIL G 818240 818241 0.20 mm –
Nano-SIL G/UV254 818342 818343 0.20 mm UV254
Nano-SIL G 818141 0.20 mm –
Nano-SIL G/UV254 818143 0.20 mm UV254
www.mn-net.com 281
Nano-DURASIL G unmodified HPTLC silica layers
· Nano silica 60, mean pore size 60 Å, specific surface (BET) · Hard, water-resistant and wettable layers due to a special
~ 500 m2/g, specific pore volume 0.75 mL/g, particle size binder system
2–10 μm · Different selectivity compared to ADAMANT and SIL-G
· Indicator: manganese activated zinc silicate with green fluo- plates no reversed phase tendency, more polar than
rescence for short-wave UV (254 nm) Nano-SIL
Glass plates
Nano-DURASIL-20 812010 812011 0.20 mm –
Nano-DURASIL-20 UV254 812013 812014 0.20 mm UV254
MACHEREY-NAGEL
CHROMABOND® SPE and Flash products
High-performance products for sample preparation

· Comprehensive range of RP- and normal phases as well as
ion exchangers
· Polymer and silica based phases
· Phases for special applications like food or environmental
analysis
· SPE polypropylene columns and cartridges,
MULTI 96 plates and SPE accessories
· High throughput SPE
· Flash chromatography cartridges
More information from page 9 onwards as well as online at www.mn-net.com/chroma
282 www.mn-net.com
Modified silica layers
Nano-SIL C18 G octadecyl-modified HPTLC silica layers
Technical characteristics Modification Recommended application

· Nano silica 60, mean pore size 60 Å, · Partial (50 %) or complete (100 %) oc- · Reversed phase separation mode
specific surface (BET) ~ 500 m2/g, tadecyl modification, carbon content with eluents from anhydrous solvents
specific pore volume 0.75 mL/g, pH 7.5 and 14 %, respectively to mixtures with high concentrations
stability 2–10, particle size 2–10 μm · Order of polarity: of water (see table and figure below)
· Indicator: acid-resistant product with a silica > DIOL > NH2 > CN > RP-2 > · Alkaloids, amino acids, preservatives,
pale blue fluorescence for short-wave C18-50 > RP-18 W > C18-100 optical brighteners, barbiturates, poly-
UV (254 nm), UV-absorbing substanc- cyclic aromatic hydrocarbons (PAH),
es appear as dark-blue to black spots drugs, peptides, flavonoids, phenols,
on a light-blue background indole derivatives, steroids
Pack of [plates] 25
Glass plates
Nano-SIL C18-50 50 % silanized 811054 0.20 mm –
Nano-SIL C18-50 UV254 50 % silanized 811064 0.20 mm UV254
Nano-SIL C18-100 100 % silanized 811052 0.20 mm –
Nano-SIL C18-100 UV254 100 % silanized 811062 0.20 mm UV254
Eluent v/v Migration distances [mm/15 min]

C18-50 C18-100 RP-18 W
Methanol – H2O 2:1 57 45 44
1:1 52 21 40
60
1:2 50 0 43
1:3 40 0 45 c)
for a distance of 7 cm
Developing time [min]
1:4 30 0 46
0:1 0 0 54
Acetonitrile – H2O 2:1 62 46 66 40 a) b)
1:1 52 30 54
1:2 51 27 46
1:3 48 15 44
1:9 20 0 42
Trichloromethane 68 64 71 20
Migration of C18-50 and C18-100 silica layers as compared to RP-18 W

plates
0
100 80 60 40 20 0 % H2O
a) RP-18 W, b) Nano-SIL C18-50, c) Nano-SIL C18-100
all plates with UV indicator
methanol – water; acetonitrile – water
Elution properties of MN RP plates in mixtures of methanol – water and
acetonitrile – water
www.mn-net.com 283
RP-18 W/UV254 G A octadecyl-modified HPTLC silica layers

· Nano silica 60, mean pore size 60 Å, · Partial octadecyl (C18) modification, · NP or RP separation with eluents
specific surface (BET) ~ 500 m2/g, wettable with water, carbon content from anhydrous solvents to mixtures
specific pore volume 0.75 mL/g, 14 % with high concentrations of water (see
particle size 2–10 μm, for prepara- · Order of polarity: table and figure on previous page),
tive plates (1 mm thickness of layer) silica > DIOL > NH2 > CN > RP-2 > relative polarity of the eluent deter-
standard silica 60, pH stability 2–10, C18-50 > RP-18 W > C18-100 mines the polarity of the layer
particle size 5–17 μm · Aminophenols, barbiturates, pre-
· Indicator: acid-resistant product with a servatives, nucleobases, polycyclic
pale blue fluorescence for short-wave aromatic hydrocarbons, steroids,
UV (254 nm), UV-absorbing substanc- tetracyclines, plasticizers (phthalates)
es appear as dark-blue to black spots
on a light-blue background
Plate size [cm] 4x8 5 x 10 5 x 20 10 x 10 10 x 20 20 x 20 Thickness of layer Fluorescent indicator
Glass plates
Pack of [plates] 50 25 50 25
RP-18 W/UV254 811073 811075 811072 811071 0.25 mm UV254
RP-18 W/UV254 811074 1.00 mm UV254
ALUGRAM aluminum sheets
®
Pack of [plates] 50 50 50 25 25
RP-18 W/UV254 818144 818152 818145 818147 818146 0.15 mm UV254
RP-2/UV254 G A “silanized silica” = dimethyl-modified standard silica layers

· Silica 60, mean pore size 60 Å, specif- · Silanized silica with dimethyl modifi- · Normal phase or reversed phase se-
ic surface (BET) ~ 500 m2/g, specific cation, carbon content 4 % paration modes with purely organic,
pore volume 0.75 mL/g, pH stability · Order of polarity: organic - aqueous or purely aqueous
2–10, particle size 5–17 μm silica > DIOL > NH2 > CN > RP-2 > eluents
· Indicator: acid-resistant product with a C18-50 > RP-18 W > C18-100 · Active plant constituents, steroids
pale blue fluorescence for short-wave
UV (254 nm), UV-absorbing substanc-
Glass plates
RP-2/UV254 811081 811082 0.25 mm UV254
RP-2/UV254 818171 0.15 mm UV254
284 www.mn-net.com
Nano-SIL CN G A cyano-modified HPTLC silica layers

· Nano silica 60, mean pore size 60 Å, · Cyanopropyl modification, carbon · NP or RP separation modes depend-
specific surface (BET) ~ 500 m2/g, content 5.5 % ing on the polarity of the developing
specific pore volume 0.75 mL/g, pH · Order of polarity: solvent (see figure below)
stability 2–8, particle size 2–10 μm silica > DIOL > NH2 > CN > RP-2 > · Steroid hormones, phenols, preser-
· Indicator: acid-resistant product with a C18-50 > RP-18 W > C18-100 vatives
pale blue fluorescence for short-wave
UV (254 nm), UV-absorbing substanc-
Rf values of different steroids as a function Separation of preservatives

of eluent composition
MN Appl. No. 401440
Layer: Nano-SIL CN/UV
Sample volume: 400 nL
Cholesterol Cortisone Eluent: ethanol – water – glacial acetic acid (20:80:0.2) with
Rf Androsterone 0.1 mol/L tetraethylammonium chloride
1.0 Migration distance: 73 mm in 30 min
Partition
Detection: TLC scanner, UV 254 nm
(reversed phase)
0.8
Peaks:
0.6 1. Propyl p-hydroxybenzoate
2. Ethyl p-hydroxybenzoate
Adsorption 3. Methyl p-hydroxybenzoate
0.4
(normal phase)
4. Benzoic acid
0.2 5. Sorbic acid
2
0.0 5
0 20 40 60 80 100 80 60 40 20 0 4
% Acetone 1 3
100 80 60 40 20 0 20 40 60 80 100
% Petroleum ether % Water
Layer: Nano-SIL CN/UV

start
Polarity of the eluent governs the type of separation mechanism:

Eluent system petroleum ether (PE) – acetone (NP mode)
the higher the concentration of PE, the stronger are the adsorptive inter-
actions of the steroids with the stationary phase
Eluent system acetone – water (RP mode)
the sequence of elution of the steroids is reversed, the most nonpolar
compounds are most strongly retained
73 mm
Plate size [cm] 4x8 10 x 10 10 x 20 Thickness of layer Fluorescent indicator
Pack of [plates] 50 25 25
Glass plates
Nano-SIL CN/UV 811115 811116 0.20 mm UV254
®
Nano-SIL CN/UV 818184 0.15 mm UV254
www.mn-net.com 285
Nano-SIL NH2 G A amino-modified HPTLC silica layers

· Nano silica 60, mean pore size 60 Å, · Aminopropyl modification, carbon · Vitamins, sugars, steroids, purine
specific surface (BET) ~ 500 m2/g, content 3.5 % derivatives, xanthines, phenols,
specific pore volume 0.75 mL/g, pH · Order of polarity: nucleotides and pesticides
stability 2–8, particle size 2–10 μm silica > DIOL > NH2 > CN > RP-2 >
· Indicator: acid-resistant product with a C18-50 > RP-18 W > C18-100
pale blue fluorescence for short-wave · Layer can be wetted equally well with
UV (254 nm), UV-absorbing substanc- pure water as with organic solvents
Influence of eluent composition Separation of sugars

on the separation of nucleotides MN Appl. No. 401590
Layer: Nano-SIL NH2/UV
Rf Sample volume: 0.5 μL
1.0 Eluent: ethyl acetate – pyridine – water – glacial acetic acid
Adenosine
c-AMP (60:30:10:5, v/v/v/v)
Migration distance: 80 mm in 45 min, double development
0.8 Detection: dry layer at 160 °C for 5 min, TLC scanner, UV
254 nm
0.6 Peaks (0.1 % each):

AMP
1. Lactose
2. Saccharose
0.4
3. Galactose
4. Glucose 1
ADP
0.2 5. Fructose
ATP 6. Arabinose
7. Xylose 2
5
0.0 8. Ribose
3 6 7
0 20 40 60 80 100 % H2O 4
100 80 60 40 20 0 % MeOH
Layer: Nano-SIL NH2/UV 8

Eluent: MeOH – H2O according to fig.
+ 0.18 mol/L NaCl
Migration distance: 7 cm
c-AMP, AMP: adenosine monophosphate

ADP: adenosine diphosphate
ATP: adenosine triphosphate 8 50 mm
Glass plates
Nano-SIL NH2/UV 811111 811112 0.20 mm UV254
®
Nano-SIL NH2/UV 818182 0.15 mm UV254
286 www.mn-net.com
Nano-SIL DIOL G diol-modified HPTLC silica layers

· Nano silica 60, mean pore size 60 Å, · Diol modification, carbon content · Steroids, pesticides and plant con-
specific surface (BET) ~ 500 m2/g, 5.5 % stituents
specific pore volume 0.75 mL/g, pH · Order of polarity: · For critical separations an alternative
stability 2–8, particle size 2–10 μm silica > DIOL > NH2 > CN > RP-2 > to silica
· Indicator: acid-resistant product with a C18-50 > RP-18 W > C18-100 · Since it is less sensitive to the water
pale blue fluorescence for short-wave · Layer can be wetted equally well with content of the environment, leads to
UV (254 nm), UV-absorbing substanc- pure water as with organic solvents more reproducible results compared
es appear as dark-blue to black spots to silica
Separation of herbicides
MN Appl. No. 401950
Layer: Nano-SIL DIOL/UV
Sample volume: 2 μL
Eluent: petroleum ether (40–60 °C) – acetone (80:20, v/v)
Migration distance: 70 mm
Detection: TLC scanner, 230 nm
Peaks:
2
(0.07 % each in methanol)
1. Metoxuron
2. Monuron 1 3
3. Metobromuron
12.0 45.3 85.0 mm
Pack of [plates] 25
Glass plates
Nano-SIL DIOL/UV 811120 0.20 mm UV254
www.mn-net.com 287
Further layers
Alox G P A aluminum oxide layers
Technical characteristics Recommended application

· Aluminum oxide, mean pore size 60 Å, specific surface (BET) · Terpenes, alkaloids, steroids, aliphatic and aromatic com-
~ 200 m2/g pounds
· Inert organic binder · We recommend to activate aluminum oxide layers before
· Indicator: manganese-activated zinc silicate use by heating 10 minutes at 120 °C
Separation of bisadducts of fullerenes Separation of lipophilic dyes

MN Appl. No. 401930 MN Appl. No. 403010
F. Djojo, A. Hirsch, Chem. Eur. J. 4 (1998), 344–356 Layer: Alox-25 UV254
Layer: ALUGRAM® Alox N/UV254 Sample volume: 1000 nL
Eluent: toluene – ethyl acetate (95:5, v/v) Eluent: toluene – cyclohexane (2:1, v/v)
2
Detection: UV, 254 nm Migration distance: 108 mm in 15 min
Detection: TLC scanner, UV 254 nm
Compound Rf values Peaks:

Bis[bis(4-phenyloxazolin)methane]fullerene 1 0.14 1. Indophenol
Bis[bis(4-phenyloxazolin)methane]fullerene 2 0.26 2. Sudan red G
3. Sudan blue II
1
4. Butter yellow
Ph Ph Ph
Ph
N N N
N
O O O
O
3
O
O
Ph
N 4
N Ph
N
O N O
1 2
Ph
Ph
0.0 25.0 50.0 75.0 100.0 125.0 mm
Glass plates
Alox-25 UV254 807021 807023 0.25 mm UV254
Alox-100 UV254 807033 1.00 mm UV254
POLYGRAM polyester sheets
®
Alox N/UV254 802021 802022 802023 0.20 mm UV254
®
Alox N/UV254 818024 818023 0.20 mm UV254
288 www.mn-net.com
Further layers
Cellulose MN 300 G P A native fibrous cellulose layers

· Fiber length (95 %) 2–20 μm, average degree of polymeriza- · Partition chromatography of polar substances such as ami-
tion 400–500, specific surface acc. to Blaine 15 000 cm2/g, no acids, carboxylic acids or carbohydrates
≤ 20 ppm Fe, 6 ppm Cu, 7 ppm P; CH2Cl2- extract ≤
0.25 %; residue on ignition at 850 °C ≤ 1500 ppm
Glass plates
Pack of [plates] 25
CEL 300-10 808013 0.10 mm –
CEL 300-10 UV254 808023 0.10 mm UV254
CEL 300-25 808033 0.25 mm –
CEL 300-25 UV254 808043 0.25 mm UV254
CEL 300-50 808053 0.50 mm –
CEL 300-50 UV254 808063 0.50 mm UV254
CEL 300 801011 801013 0.10 mm –
CEL 300 UV254 801022 801023 0.10 mm UV254
CEL 300 818155 818153 0.10 mm –
CEL 300 UV254 818157 818156 0.10 mm UV254
Cellulose MN 400 (AVICEL®) G P microcrystalline cellulose layers

· Prepared by hydrolysis of high purity cellulose with HCl, · Carboxylic acids, lower alcohols, urea and purine deriva-
average degree of polymerization 40–200 tives
Glass plates
CEL 400-10 808072 808073 0.10 mm –
POLYGRAM polyester sheets
®
CEL 400 801113 0.10 mm –

CEL 400 UV254 801123 0.10 mm UV254
www.mn-net.com 289
Further layers
Cellulose MN 300 PEI P PEI-impregnated cellulose ion exchange layers

· Fibrous cellulose impregnated with polyethyleneimine · Analysis of nucleic acids, and of mutagenic substances with
the 32P postlabelling procedure
Pack of [plates] 25
CEL 300 PEI 801053 0.10 mm –
CEL 300 PEI/UV254 801063 0.10 mm UV254
Cellulose MN 300 AC P acetylated cellulose layers

· Fibrous cellulose with 10 % content of acetylated cellulose · Reversed phase chromatography
for reversed phase chromatography
Plate size [cm] Acetyl content 20 x 20 Thickness of layer Fluorescent indicator
Pack of [plates] 25
CEL 300 AC-10 % 10 % 801033 0.10 mm –
Polyamid-6 P ε-polycaprolactame layers

· Polyamide 6 = nylon 6 = perlon = ε-aminopolycaprolactame · Natural compounds, phenols, carboxylic acids, aromatic
· Separation mechanism based on hydrogen bonds to amide nitro compounds and especially amino acids
groups of the polymer matrix as well as on ionic, dipole and
electron donor-acceptor interactions
POLYAMID-6 803012 803013 0.10 mm –
POLYAMID-6 UV254 803022 803023 0.10 mm UV254
290 www.mn-net.com
Layers for special TLC separations
CHIRALPLATE G special layer enantiomer separation

· Reversed phase nano silica impregnated with Cu2+ ions and · Enantiomer separation of amino acids, N-methylamino
a chiral selector (proline derivative) acids, N-formylamino acids, α-alkylamino acids, thiazolidine
· Separation based on ligand exchange, i.e. formation of derivatives, dipeptides, lactones, α-hydroxycarboxylic acids
ternary mixed-ligand complexes with the Cu(II) ions, differ-
ences in the stability of the diastereomeric complexes cause
chromatographic separation
Enantiomer separation of amino acids

MN Appl. No. 400520
Quantitative determination (remission location curves) of TLC-separated enantiomers of tert.-leucine:
Layer: CHIRALPLATE a) b) c) d)
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
Plate size [cm] 5 x 20 10 x 10 10 x 20 20 x 20 Thickness of layer Fluorescent indicator
Glass plates
Pack of [plates] 4
CHIRALPLATE 811056 0.25 mm UV254
Pack of [plates] 50 25 25 25
CHIRALPLATE 811057 811059 811055 811058 0.25 mm UV254
SIL N-HR P unmodified standard silica layers
· High purity silica 60, mean pore size 60 Å, specific surface · A special feature of the POLYGRAM® SIL N-HR is a higher
(BET) ~ 500 m2/g, specific pore volume 0.75 mL/g, particle gypsum content
size 5–17 µm, different binder system compared to SIL G
results in different separation characteristics
SIL N-HR/UV254 804022 804023 0.20 mm UV254
www.mn-net.com 291
SIL G-25 HR G special layer for aflatoxin separation

· High purity silica 60 with gypsum and a very small quantity · Aflatoxins
of a polymeric organic binder; softer than the standard silica
layer, i.e. spots can be scratched and the layer absorbs
faster
Pack of [plates] 25
Glass plates
SIL G-25 HR 809033 0.25 mm –
SIL G-25 HR/UV254 809043 0.25 mm UV254
SIL G-25 Tenside G special layer for separation of surfactants

· Silica G impregnated with ammonium sulfate · Detergents, alkanesulfonates, polyglycols
Pack of [plates] 25
Glass plates
SIL G-25 Tenside 810063 0.25 mm –
Nano-SIL PAH G special HPTLC silica layer for PAH analysis

· Nano silica 60, mean pore size 60 Å, specific surface (BET) · 6 PAHs according to German drinking water specifications
~ 500 m2/g, specific pore volume 0.75 mL/g, particle size (TVO) in accordance with German standard DIN 38407
2–10 μm part 7
· Impregnated with caffeine, an electron acceptor for PAH
analysis based on charge-transfer complexes
Pack of [plates] 50
Glass plates
Nano-SIL PAH 811051 0.20 mm –
292 www.mn-net.com
IONEX P special mixed layers of silica with ion exchange resins
IONEX-25 SA-Na: Recommended application

· Mixture of silica and a strongly acidic cation exchanger coat- · Amino acids, e.g., in protein and peptide hydrolyzates, in
ed to polyester sheets seeds and fodder, in biological fluids; for racemate separa-
IONEX-25 SB-AC: tion in peptide syntheses, for the separation of nucleic
acid hydrolyzates, aminosugars, amino acids, antibiotics,
· Mixture of silica and a strongly basic anion exchanger coat-
inorganic phosphates, cations and other compounds with
ed to polyester sheets
ionic groups
· Both layers contain an inert organic binder
Pack of [plates] 25
IONEX-25 SA-Na strongly acidic cation exchanger 806013 0.20 mm –
IONEX-25 SB-AC strongly basic anion exchanger 806023 0.20 mm –
Mixed layers for TLC G
Alox/CEL-AC-Mix-25: SILCEL-Mix-25:
· Mixed layer of aluminum oxide G and acetylated cellulose, · Mixed layer of cellulose and silica, recommended for sepa-
recommended for separation of PAH ration of preservatives and other antimicrobial compounds
Pack of [plates] 25
Glass plates
Alox/CEL-AC-Mix-25 810053 0.25 mm –
SILCEL-Mix-25 UV254 810043 0.25 mm UV254
www.mn-net.com 293
Chromatography papers
Chromatography papers
Chromatography papers Please note Direction
· Paper chromatography is the oldest · Always treat chromatography papers · Chromatography papers possess a
chromatographic technique separa- with care preferred direction of the fibers with
tion due to partition of the analytes · Never touch them with fingers, higher absorption properties (with our
between special paper grades and the because this will contaminate the sheets 58 x 60 cm, the longer edge)
mobile phase, which penetrates the surface · We recommend to use them in the
paper by capillary action ascending. direction of higher absorption
· Do not bend them sharply, because
· Descending and circular techniques this will decrease the capillary action
are possible (preferably store them flat)
Code Weight [g/m2] Thickness [mm] Description Flow rate Size [cm] Pack of REF
MN 214 140 0.28 smooth 90–100 mm/30 min 58 x 60 100 sheets 817001
MN 827 270 0.70 soft carton 130–140 mm/10 min 58 x 60 100 sheets 817005
MN 214 ff 140 0.28 MN 214 defatted * 90–100 mm/30 min 56 x 58 100 sheets 817008
* This paper is extracted with organic solvents.
For further papers, filters and membranes, feel free to ask for our catalog “Filtration”.
294 www.mn-net.com
Accessories
Accessories
· Beside ready-to-use layers for thin layer chromatography · Selection of accessories for reliable separation in TLC
also accessories are required
Simultaneous developing chamber for TLC, 20 x 20 cm 1 814019
Simultaneous developing chamber for TLC, 10 x 10 cm 1 814018
Developing chambers for TLC micro-sets 4 814021
Glass laboratory sprayer with rubber bulb 1 814101
Glass capillaries 1 μL 3 x 50 814022
Rubber caps for capillaries 2 814102
Plastic syringe, 1 mL content with graduation 1 814104
Spotting guides 2 814023
Measuring cylinders, glass, 10 mL content 2 814024
MN ALUGRAM® scissors, ground blade, black handle 1 818666
Filter paper MN 713, 15 x 21 cm 100 814103
Folded filters MN 615 1/4, 11 cm diameter 100 531011
Chromatography paper MN 260, 7.5 x 17 cm (for chamber saturation) 100 814030
www.mn-net.com 295
Reagents
Visualization reagents
· Small selection of frequently used spray reagents for post · A detailed description of many more detection procedures
chromatographic detection reactions in TLC suited for spray- for TLC is available on request
ing or dipping TLC plates
Spray reagent Solvent Detection of Pack of REF
Aniline phthalate 2-propanol – ethanol (1:1) reducing sugars, oxohalic acids 100 mL 814919
Bromocresol green 2-propanol organic acids 100 mL 814920
Reagent for caffeine detection water – acetone caffeine 100 mL 814401
2’,7’-Dichlorofluorescein 2-propanol lipids (saturated, unsaturated) 100 mL 814921
4-(Dimethylamino)-benzaldehyde 2-propanol terpenes, sugars, steroids 100 mL 814922
Reagent according to water alkaloids and other nitrogen compounds 100 mL 814402
Dragendorff-Munier
Iron(III) chloride water phenolic compounds e.g., acetylsalicylic acid, para- 100 mL 814403
Potassium hexacyanoferrate(III) water cetamol 100 mL 814404
Molybdatophosphoric acid ethanol lipids, sterols, steroids, reducing compounds 100 mL 814302
Ninhydrin ethanol amino acids, amines and amino sugars 100 mL 814203
Rhodamine B ethanol lipids 100 mL 814923
Rubeanic acid ethanol heavy metal cations 100 mL 814206
These products contain harmful substances which must be specially labeled as hazardous. For detailed information please see SDS.
Fluorescent indicators
UV indicators with efficient radiation for short-wave as well as long-wave UV ranges
· UV254: manganese-activated zinc silicate with absorption · UV366: inorganic fluorescent pigment with absorption maxi-
maximum at 254 nm, green fluorescence, relatively sus- mum at 366 nm, blue fluorescence
ceptible towards acids: its fluorescence can be completely
quenched by acidic solvents
Composition Absorption maximum Color of fluorescence Pack of 100 g
Fluorescent indicator UV254 manganese-activated zinc silicate 254 nm green 816710.01
Fluorescent indicator UV366 inorganic fluorescent pigment 366 nm blue 816720.01
296 www.mn-net.com
Adsorbents
Silica adsorbent for TLC

Pore size 60 Å, pore volume 0.75 mL/g, specific surface (BET) ~ 500 m2/g,
pH 7 for a 10 % aqueous suspension
· Silica G: standard grade, particle size 2–20 μm, Fe < 0.02 %, · Silica P: preparative grade, particle size 5–50 μm,
Cl < 0.02 %, 13 % gypsum as binder Fe < 0.02 %, Cl < 0.02 %, organic binder
· Silica N: standard grade, particle size 2–20 μm, Fe < 0.02 %, · Silica P with gypsum: preparative grade, particle size
Cl < 0.02 %, no binder 5–50 μm, Fe < 0.02 %, Cl < 0.02 %, gypsum as binder
· Silica G-HR: high purity grade, particle size 3–20 μm,
Fe < 0.002 %, Cl < 0.008 %, gypsum as binder
Designation Fluorescent indicator 1 kg 5 kg
Silica G – 816310.1 816310.5
Silica G/UV254 UV254 816320.1 816320.5
Silica N – 816330.1 816330.5
Silica N/UV254 UV254 816340.1 816340.5
Silica G-HR – 816410.1 816410.5
Silica P/UV254 UV254 816380.1 816380.5
Silica P/UV254 with gypsums UV254 816400.1 816400.5
Polyamid adsorbent for TLC

Polyamide 6 = nylon 6 = perlon = ε-polycaprolactame
Designation Fluorescent indicator 1 kg
Polyamid-DC 6 – 816610.1
Polyamid-DC 6 UV254 UV254 816620.1
Cellulose MN 301 native fibrous cellulose

· Standard grade, fiber length (95 %) 2–20 μm · ≤ 20 ppm Fe, 6 ppm Cu, 7 ppm P, CH2Cl2 extract
· Average degree of polymerization 400–500, specific surface ≤ 0.25 %, residue on ignition at 850 °C ≤ 1500 ppm
acc. to Blaine 15 000 cm2/g
Designation 1 kg 5 kg
Cellulose MN 301 816250.1 816250.5
www.mn-net.com 297

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Thin layer chromatography

The success of thin layer chromatography as a highly efficient

It is important to pay attention to the atmosphere in the de-

Evaluation of a thin layer chromatogram

Identification of separated substances is possible via the Rf val-

TLC micro-sets introductory kits for science education

TLC micro-set A for beginners Contents of TLC micro-set A for beginners

TLC micro-set F2 Contents of TLC micro-set F2

TLC micro-set F3 Contents of TLC micro-set F3

TLC micro-set M Contents of TLC micro-set M (materials kit)

Supports for ready-to-use layers for TLC

Glass plates G POLYGRAM® P ALUGRAM® A / ALUGRAM® Xtra Ax

SIL G G P A Ax silica 60, standard grade 276

DURASIL G silica 60, special binder system 277

SILGUR G Ax silica 60 with kieselguhr concentrating zone 279

Unmodified silica for HPTLC particle size 2–10 µm

Nano-SIL G A Ax nano silica 60, standard grade 281

Nano-DURASIL G nano silica 60, special binder system 282

Modified silica for HPTLC particle size 2–10 µm

RP-2/UV254 G A silanized silica = dimethyl-modified nano silica 60 284

Nano-SIL CN G A cyano-modified nano silica 285

Nano-SIL NH2 G A amino-modified nano silica 286

Nano-SIL DIOL G diol-modified nano silica 287

Cellulose, unmodified and modified

CEL 400 G P microcrystalline cellulose MN 400 (AVICEL®) 289

CEL 300 PEI P polyethyleneimine-impregnated cellulose ion exchanger 290

CEL 300 AC P acetylated cellulose MN 300 290

Layers for special separations

SILCEL-Mix G mixed layer of cellulose and silica 293

ADAMANT G unmodified standard silica layers

Key features Technical characteristics

Separation of steroids Separation of barbiturates

Substance Rf ADAMANT Rf SIL G Substance Rf

ALUGRAM® Xtra SIL G Ax unmodified standard silica layers on aluminum

Key features Technical characteristics

Separation of nutmeg ingredients

The chromatograms show the following zones with increasing Rf values:

SIL G G P A unmodified standard silica layers

DURASIL G unmodified standard silica layers

MN TLC pre-coated layers

Arbitrary application Applied substances Applied substances Developed

SILGUR G Ax unmodified standard silica layers with concentrating zone

Nano-SILGUR G Ax unmodified HPTLC silica layers with concentrating zone

Nano silica for HPTLC

Comparison of ADAMANT and Nano-ADAMANT plates for separation of anthraquinone dyes

Nano-ADAMANT G unmodified HPTLC silica layers

Key features Technical characteristics

Nano-SIL G Ax A unmodified HPTLC silica layers

Nano-DURASIL G unmodified HPTLC silica layers

High-performance products for sample preparation

More information from page 9 onwards as well as online at www.mn-net.com/chroma

Nano-SIL C18 G octadecyl-modified HPTLC silica layers

Technical characteristics Modification Recommended application

Eluent v/v Migration distances [mm/15 min]

Migration of C18-50 and C18-100 silica layers as compared to RP-18 W

RP-18 W/UV254 G A octadecyl-modified HPTLC silica layers

Technical characteristics Modification Recommended application

RP-2/UV254 G A “silanized silica” = dimethyl-modified standard silica layers

Technical characteristics Modification Recommended application