CAMAG

TLC SCANNER 4

WORLD LEADER IN PLANAR

CHROMATOGRAPHY
The TLC Scanner 4 is the most advanced workstation All functions of the scanner are controlled by the software winCATS.
for densitometric evaluation of TLC/HPTLC Only the positioning of the object to be scanned is performed manu-
ally. If desired the operator can switch on the internal illumination to
chromatograms. It can also be used for densitometric control or adjust the scan start position.
measurements of other planar objects. Optimal settings for electronic amplification are automatically selected
for measurements in absorption and fluorescence mode.

Perfect evaluation with winCATS

The well structured and easy to use winCATS software controls and
monitors all functions of the scanner and processes data up to the
final result.
The winCATS standard program for the TLC Scanner 4 features:
s Short measurement times
s Measurement of up to 36 tracks with up to 100 substances per track
s Integration either with automatic or manual baseline correction /
peak assignment
s Automatic or easy manual assignment of substance names to peaks
s Automatic recording of spectra of all detected or selected peaks
s Printout or export of color graphics of measurement data
s Report printout of the entire analysis including all measurement
data and images of the TLC/HPTLC plate
A number of winCATS program options allow the user to fit For quantitation, data from the optimum wavelength scan of each
the evaluation system to the respective need. The following fraction can be selected. This software function is unique in Thin-
options are available: Layer Chromatography!
s Quantitative evaluation s Scanner qualification (Selftest)
supports the following calibration modes: single level calib- This option offers automatic monitoring of the mechanical, opti-
ration, multi level calibration with linear or nonlinear regres- cal and electronic functions of the scanner. Results are evaluated,
sion using internal or external standards. Statistics are documented and saved. If appropriate, lamp positions and mono-
available as relative standard deviation (cv) or confidence chromator alignment can be automatically adjusted.
interval (ci). s Track optimization
s Sub-component evaluation Each track of a chromatogram is scanned several times with small
(included in the option “Quantitative Evaluation”) can be used lateral offset. From this data the optimum track following the peak
to quantify unknown peaks by relating them to the main com- maxima is calculated and used for quantitation. Thereby distorted
ponent as is prescribed by European or US pharmacopoeias chromatograms can be correctly evaluated.
(“Related Compounds”). s Spectrum library
s Dual-wavelength scan Enables the user to create his own library files, which provides
The chromatogram is scanned at two individually selectable wave- the only way the user can save and compare spectra recorded on
lengths. During integration the signal from the second wavelength different plates.
is subtracted from that of the first wavelength to eliminate matrix
effects. Dual-wavelength scanning is also useful for the quantita-

I
tion of incompletely resolved peaks.
Note
s Multi-wavelength scan
The chromatogram can be scanned automatically at up to The TLC Scanner 4 with winCATS is compliant with the requirements
36 different wavelengths. This permits multiple measurements be- of GMP/GLP and can be IQ/OQ qualified.
tween 190 and 900 nm in order to achieve optimum selectivity. If you want to use the instrument in a 21 CFR Part 11 environment,
the option 21 CFR Part 11 “compliance ready” is required for each
winCATS workstation.

CAMAG TLC Scanner 4 2|3

CAMAG TLC SCANNER 4

Key features
s Measurement of reflection, either in absorbance or fluorescence
mode
s Object formats up to 200 × 200 mm
s Spectral range from 190 to 900 nm
s Automatic start of all lamps: deuterium, halogen-tungsten, and
high pressure mercury lamp
s Data step resolution 25 – 200 µm
s Scanning speed 1 – 100 mm/s Planar chromatogram
s Spectra recording speed up to 100 nm/s
s Automatic selection of electronic amplification
s Rapid data transfer

Analog curve

Calibration function
User-friendly operation I
All functions of the scanner are controlled by winCATS software.

The electronic amplification is set automatically. In fluorescence mode this is performed by a

“quick scan”, which measures the complete plate, e.g. 18 tracks in 20 seconds, and determines
the optimized settings from the result. In absorbance mode only one track is “quick scanned”;
either track 1 (automatic mode) or a track of choice (manual mode).
The 16 bit A/D converter ensures optimum resolution of the signal. This is particularly useful
for the measurement of both low absorbance and weak fluorescence signals.
Scanner operation with winCATS software is convenient and easy to learn.

The object, here a 20 × 10 cm HPTLC plate, is

simply positioned on the scanning table.

The coordinates are displayed during manual

positioning of the stage and can be transfer-
red into the program by mouse-click.

CAMAG TLC Scanner 4 4|5

 Lamp selector Entrance lens Monochromator Monochromator grating
system entry slit


THE OPTICAL SYSTEM:

Mirror
FEATURES AND TECHNICAL DETAILS

The Optical System Light sources

s Any of the three light sources, high pressure mercury lamp, deute- s Deuterium lamp, usable continuum 190 – 450 nm
rium lamp, or halogen-tungsten lamp can be positioned in the light s Halogen-tungsten lamp, usable continuum 350 – 900 nm
path by a motor drive. 
s High-pressure mercury lamp, line spectrum 254 – 578 nm
s The signal of the measuring photomultiplier is continuously offset
against the signal of the reference photomultiplier.   The lamp, which is positioned in the light path, is automatically ignited.
This compensates for lamp aging and short-time fluctuations. It also All lamps are current stabilized.
reduces the warm-up time required to reach lamp stabilization.
s All components of the optical system, lamps, monochromator, scan- Pilot lamp and compartment illumination
ning stage, and photomultiplier are mounted on one sturdy metal The slit is automatically illuminated with visible light when the com-
support. This ensures high precision of the detector signal. partment illumination is switched on. The scanning compartment is
s For scanning at wavelengths below 200 nm it is advisable to flush illuminated with a 4 watt fluorescent tube UV 254 nm which the user
the monochromator with nitrogen. The scanner is equipped to do can replace by a UV 366 nm or a white light tube.
this.
s A monochromator bandwidth of 5 nm or 20 nm can be selected. Optical system
5 nm bandwidth is used for spectra recording, multi-wavelength Apochromatic suprasil-fluorite lens system, transmission range
scanning, and always when spectral selectivity is required.  190 – 900 nm, astigmatic entry lens for optimal slit illumination;
20 nm bandwidth offers higher light intensity (improves the signal- automatic switching between micro and macro position for optimal
to-noise ratio and thus the reproducibility of the measurement) and light intensity.
enables measurement of several fractions with slightly different
absorption maxima in one scan.
Monochromator
s The lens system with 190 – 900 nm transmission range features
automatic positioning for micro and macro slit sizes. This ensures Concave holographic grating, 1200 lines/mm, bandwidth selectable 5
that the light energy available with small slits in the micro position or 20 nm, wavelength range 190 – 900 nm; monochromator driven by
is almost the same as that for the corresponding slit in the macro stepper motor, reproducibility of wavelength setting better than 0.2 nm,
position, which is four times larger.  accuracy better than 1 nm; connector for flushing with nitrogen.

s The light beam strikes the object at right angle. The photomultiplier Maximum speed of spectra recording 100 nm/s, positioning at
is aligned at an angle of 30°.  200 nm/s.
Disk with slit Lens system, can
apertures be positioned
for micro and macro Mirror
slit image

Reference
photomultiplier

Beam splitter

 Measuring
photomultiplier

Scanning object

Secondary filter Mains voltage

Motor-driven filter wheel with three automatically selected filters for 115 V and 230 V selectable; 50/60 Hz; maximum energy use 180 W
the elimination of second order wavelengths; 400 nm cut-off filter for (tungsten and mercury lamp ignited).
fluorescence measurements; three positions for user selected filters.
A/D converter
Scanning slit 16 bit, 2-channel A/D converter, 100 ms per double conversion.
Revolving disk with 20 fixed apertures; length of slit images selec-
table between 0.2 and 12 mm, width between 0.1 and 1.2 mm in Connections/interfaces
42 combinations.
Serial interface RS232 for communication to the computer, EquiLink
for connection to winCATS software.
Detector
Two matched broad band photo multipliers, multi alkali type, spectral Dimensions
sensitivity 185 – 900 nm.
Width = 590 mm, depth = 650 mm, height = 367 mm; net weight 39 kg.

Stage drive
Independent in both directions by stepper motors, micro step dri-
ven for smooth movement; reproducibility of positioning better than
50 µm in Y-direction, better than 100 µm in X-direction; maximum
scanning speed 100 mm/s, positioning at 150 mm/s.

CAMAG TLC Scanner 4 6|7

 Manual Input Horizontal
Developing Chamber Immersion Device

Stationary Phase Nanomat Developing Chamber Spray

Sample Sample Chromatogram

Preparation Definition Application Development Derivatization

Automatic TLC Automated Multiple

Samples Sampler ATS 4 Development AMD 2
Automatic Developing
Software Controlled Standards Linomat 5 Chamber ADC 2

winCATS SOFTWARE

winCATS for all steps of instrumental winCATS features

Thin-Layer Chromatography The winCATS standard program for the TLC Scanner 4 supports:
CAMAG has been developing software for densitometric evaluation
of TLC/HPTLC chromatograms since 1980. s Short measurement times
s Measurement of up to 36 tracks with up to 100 substances per track
winCATS is the result of a unique integrated software concept s Integration either with automatic or manual baseline correction /
covering all steps of Instrumental Thin-Layer Chromatography. The peak assignment
user can combine the individual modules to create a complete solu-
tion that meets all requirements with respect to instrument control, s Automatic or easy manual assignment of substance names to peaks
data acquisition, evaluation and documentation. s Automatic recording of spectra of all detected or selected peaks
The modular design of winCATS allows selecting or disabling the in- s Printout or export of color graphics of measurement data
dividual steps of Thin-Layer Chromatography according to the task s Report printout of the entire analysis including all measurement
at hand. data and images of the TLC/HPTLC plate
The program offers a unique combination of features:
s winCATS combines high performance with easy handling
s winCATS offers a two-stage on-screen help system, a brief info line
and a detailed explanatory help text
s winCATS features a structured data management and high data
safety with easy access to network and project directories
s winCATS stores the complete set of parameters used together with
all data – from scanning raw data to documentation – in one single
file, and prints everything if desired
s winCATS complies with the rules of GMP/GLP and 21 CFR part 11
 Image Import

Image Annotation

Detection Evaluation Documentation Report

21 CFR Part 11
Quantitative
TLC Visualizer (FDA Regulation)
TLC Scanner Evaluation Image
Enhancement
Scanner Selftest Spectrum
Image
Dual-Wavelength Scan Library Comparison

Multi-Wavelength Scan
Track Optimization

Modular design Workflow

The modular design of winCATS allows selecting or disabling Using the explorer type view the operator at all times remains
the individual steps of Thin-Layer Chromatography according in control over all active steps of the workflow. These are:
to the task at hand.

s Selection of plate material and its pre-treatment

s Definition of samples, standards, and the calibration method
if applicable
s Sample application
s Chromatogram development
s Derivatization (pre and/or post chromatographic)
s Detection
s Spectra recording
s Quantitative evaluation
s Documentation

winCATS program options for TLC Scanner 4 I

Quantitative evaluation page 12 Multi-wavelength scan page 15
Spectrum library page 14 Scanner selftest page 16
Track optimization page 14 21 CFR Part 11 “compliance ready” page 16
Dual-wavelength scan page 15

CAMAG TLC Scanner 4 8|9

TLC SCANNER 4 AND winCATS

Data input Measurement

For routine analysis winCATS starts off with a method file con- The TLC Scanner 4 scans the plate and transfers raw data back
taining all relevant data for the current task such as instruments, to winCATS where baseline correction and peak recognition is
parameters, etc. This method can be validated for GxP work. performed automatically in the background.

From this method an analysis file is generated and parameters unique

to the actual analysis, i.e. sample names, amounts, etc. are adapted.
After all steps in the method leading up to densitometric evaluation
are completed, the plate is positioned on the stage and scanning is
started.
Slit dimensions, scanning speed, light source, wavelength, etc. have
already been defined within the winCATS method.

A display of integrated peaks with substance names. This display mode ena-
bles manual baseline correction and peak integration by mouse-click.

Data input of scan parameters Spectra list

Evaluation Recording of spectra
By clicking the right mouse key on an analog curve the display winCATS can automatically record spectra as soon as all peak
can be swivelled and tilted, and additional information can be positions are known. Spectra can be displayed individually or
displayed. overlaid in one diagram.

Analog curves can be displayed either individually or in a 3D diagram. Spectra can be measured from 190 to 900 nm. If the emission range of
Both can be printed in color together with the analysis report. the deuterium lamp is exceeded, the scanner automatically switches
to the halogen-tungsten lamp. Both lamps remain ignited.

All 3D diagrams can be swivelled and tilted for easy evaluation.

Excerpts from a winCATS report (can be personalized by user)

CAMAG TLC Scanner 4 10 | 11

QUANTITATIVE EVALUATION

Quantitative evaluation is based on the comparison of peak Flexible input screens for calibration data
heights or peak areas of the unknowns with those of calibra- winCATS offers a wide choice for data input: from entering “amount
tion standards chromatographed on the same plate. per fraction” to entering amounts from balance readings or from
stock solutions.
Depending on the task at hand single-level or multi-level calibration According to the European and the US Pharmacopoeia, related com-
can be selected. pounds can be quantified by comparison with small amounts of the
main component in case their identification is not required. This main-/
Calibration modes subcomponent evaluation is part of the standard quantitative evalu-
s Single-level calibration is suitable for analyses where the concentra- ation program.
tion of the unknowns shall be checked within narrow limits. Single-
level calibration requires reduced calibration efforts. Identity and purity checking by spectra comparison
s Multi-level calibration is used when the target values are expec- The software option quantitative evaluation includes identity and pu-
ted in a comparatively wide range. winCATS offers a choice of four rity checking of substances by spectra comparison. For identity che-
calibration modes , the selection is made based on which mode cking spectra recorded at the peak maxima are compared with those
gave the best result during method validation. of standard substances. The user can define a limit for the correlation
Linear regression is a useful function when the calibration range coefficient, or let winCATS calculate identities by statistical criteria.
is narrow or the absolute amounts per fraction are small, which is For purity checking spectra are recorded at peak maximum and at
often the case by fluorescence scanning. both slopes. These spectra are then compared by winCATS either to
match user specifications or according to statistical criteria.
Non-linear regression becomes necessary when a wider calibration
range is needed and/or the absolute amounts per fraction are high.
Polynomial regression is suitable for calibration over a wide
concentration range.
Michaelis-Menten regression is suitable for calibration over a
wide concentration range and for high amounts of substances. The
Michaelis-Menten function type 1 passes through the origin. In ca-
ses where this behavior is not suitable, Michaelis-Menten type 2 may
be chosen.

Ordering information I
027.6315 Quantitative chromatogram evaluation
Application example
Analysis of 25 water-soluble food dyes in food

HPTLC is the most effective, rapid and cost-efficient solution for

the analysis of water-soluble food dyes in food. Up to 20 runs
can be chromatographed under identical conditions (by use of
the Automatic Developing Chamber ADC2) within 12 minutes
(see CBS 105 or www.camag.com).

25 food dyes (split in 3 mixtures, application volumes of 1, 2, and 3 µL)

and 6 food samples (application volumes of 2 µL; energy drink (ED), Multi-wavelength scan of mixture 1 (superposition of absorption
yoghurt (Jog), fruit drink (FD), bakery ink formulation (BI)) curves at 6 wavelengths).

Calibration function of the red food dye E122 (x) and determination in Calibration function of the blue food dye E131 (x) and determination
fruit drink, energy drink and bakery ink formulation (+). in the yoghurt sample (+).

CAMAG TLC Scanner 4 12 | 13

SPECTRUM LIBRARY TRACK OPTIMIZATION

Recording, displaying and comparing spectra of substances With this software option a distorted chromatogram can be
chromatographed on one plate is included in the standard corrected.
Scanner software.
Each track is scanned several times with small lateral offset. The num-
In order to compare spectra of substances chromatographed on dif- ber of scans and the distance in between is selected according to the
ferent plates or to compare spectra with those of a spectra collection, requirements of the particular chromatogram. When all scans of one
the software option spectrum library is required. track are completed, the software selects the maximum signal for all
The spectrum library can be used during substance assignment for peaks. Only those data are then used for the calculation of the result
validation of assigned substances as well as for identification of from the corrected chromatogram.
unknown fractions.
Note:
During the search process for identification the spectrum library shows
a hit list of the closest substances including their spectra. s When used with chromatograms with proper track alignment, the results
obtained with or without track optimization are about the same
s In case of distorted chromatograms, results obtained with track
optimization are comparable to those of a good chromatogram
scanned without track optimization

Ordering information I Ordering information I

027.6342 Spectrum library 027.6344 Track optimization
DUAL-WAVELENGTH SCAN AND MULTI-WAVELENGTH SCAN

The software option dual-wavelength scan serves for back- The option multi-wavelength scan is a useful tool for the
ground correction. The chromatogram is scanned with two quantitation of analyte mixtures with components differing in
wavelengths, the measuring and the reference wavelength, absorption maxima.
which each can be selected between 190 and 900 nm.
The plate tracks can be scanned consecutively at up to 36 different
As measuring wavelength one will usually select the wavelength wavelengths between 190 and 900 nm and the peak data is stored
of the absorption maximum of the substance to be calibrated. The in one analysis file. During the consecutive quantitative evaluation
reference wavelength should be sufficiently apart from this maximum each component can now be automatically evaluated at its maximum
absorbance in order to achieve good sensitivity, however, not too absorbance. This way the user is relieved from manually evaluating
far away as otherwise irregularities in the layer will not be reliably the same plate at different wavelengths.
compensated. A preceding recording of the spectrum is helpful for For identity checking the 3D view of the multi-wavelength scan is
selecting the right combination of wavelengths. another very helpful tool. The 3D views can be scaled, swivelled and
tilted, and then copied to clipboard or saved as a bitmap file for use
in other software, e.g. Word.

Ordering information I Ordering information I

027.6346 Dual-wavelength scan 027.6348 Multi-wavelength scan

CAMAG TLC Scanner 4 14 | 15

 21 CFR PART 11
SCANNER SELFTEST “COMPLIANCE READY”

With this option an automatic selftest of the TLC Scanner 4 and This option is required for compliance with the FDA regulation
a report can be generated. regarding the recording of electronic data and signatures.

In the scanner selftest procedure the following checks are Features :

performed: s Safety of data acquisition and operation by user identification with
s Wavelength accuracy of the monochromator password
s Stage positioning s Secure storage of all results including raw data ensures complete
s Condition and alignment of all lamps data integrity and tracking

s Condition and alignment of the optical system s Documentation of all activities in History Log/Audit Trial for se-
cure tracking of electronic signatures according to 21 CFR Part 11
s Condition of the electronic system requirements
The complete qualification procedure can be carried out automatically
or manually in sections. Certain deficiencies in lamp alignment and
monochromator adjustment can be automatically corrected.

Example
The stage with the test pattern (straight line) is moved multiple
times at 0.1 mm increments in Y direction across the slit. The re-
sulting analog curve is evaluated by the program. This test gives
information about:
s Uniformity of slit illumination with the respective lamp, and
s Correct alignment of the lamp and the optical system

The result of each test is displayed on screen and can be printed as

part of the qualification report. The report contains the target values
and the values actually found together with a pass/fail judgment.
The option scanner selftest is required when the instrument shall be
operated in a GxP environment.

Ordering information I Ordering information I

027.6340 Scanner selftest 027.6380 21 CFR Part 11 “compliance ready”
INSTRUMENT QUALIFICATION INSTRUMENT SERVICE

For customers working in a GMP/GLP environment, CAMAG For all CAMAG instruments for which we offer IQ/OQ qualifi-
offers Installation Qualification (IQ) and Operation Qualifica- cation we also offer service contracts.
tion (OQ) as service.

A service contract includes:

The Installation Qualification (IQ) is performed at the site and at the
time of installation. It documents that all specifications and parame- s Preventive maintenance once per year or as required
ters comply with the manufacturer’s specifications, environmental s Adjustments and tests
parameters and safety requirements. s Optional performance of the qualification (OQ) at the same time
The Operation Qualification (OQ) is initially performed subsequent to
IQ and is repeated at certain intervals recommended by the manu-
facturer or defined by the customer. It documents that all modules
of the qualified system function properly within the specified ope-
rating ranges.
The Performance Qualification (PQ) is an ongoing process which docu-
ments that the instrument or system is suitable for the given task. Thus
only the user can perform PQ, employing his substances, following
his task description and his test procedures (SOPs).

CAMAG TLC Scanner 4 16 | 17

TECHNICAL DATA

Light sources Scanning slit

s Deuterium lamp, usable continuum 190 – 450 nm Revolving disk with 20 fixed apertures; length of slit images selec-
s Halogen-tungsten lamp, usable continuum 350 – 900 nm table between 0.2 and 12 mm, width between 0.1 and 1.2 mm in
s High-pressure mercury lamp, line spectrum 254 – 578 nm 42 combinations.
The lamp, which is positioned in the light path, is automatically ignited.
All lamps are current stabilized. Detector
Two matched broad band photo multipliers, multi alkali type, spectral
Pilot lamp and compartment illumination sensitivity 185 – 900 nm.
The slit is automatically illuminated with visible light when the com-
partment illumination is switched on. The scanning compartment is Stage drive
illuminated with a 4 watt fluorescent tube UV 254 nm which the user Independent in both directions by stepper motors, micro step driven
can replace by a UV 366 nm or a white light tube. for smooth movement; reproducibility of positioning better than 50
µm in Y-direction, better than 100 µm in X-direction; maximum scan-
Optical system ning speed 100 mm/s, positioning at 150 mm/s.
Apochromatic suprasil-fluorite lens system, transmission range
190 – 900 nm, astigmatic entry lens for optimal slit illumination; auto- Mains voltage
matic switching between micro and macro position for optimal light 115 V and 230 V selectable; 50/60 Hz; maximum energy use 180 W
intensity. (tungsten and mercury lamp ignited).

Monochromator A/D converter

Concave holographic grating, 1200 lines/mm, bandwidth selectable 5 16 bit, 2-channel A/D converter, 100 ms per double conversion.
or 20 nm, wavelength range 190 – 900 nm; monochromator driven by
stepper motor, reproducibility of wavelength setting better than 0.2 nm,
accuracy better than 1 nm; connector for flushing with nitrogen. Maxi- Connections/interfaces
mum speed of spectra recording 100 nm/s, positioning at 200 nm/s. Serial interface RS232 for communication to the computer, EquiLink
for connection to winCATS software.
Secondary filter
Motor-driven filter wheel with three automatically selected filters for Dimensions
the elimination of second order wavelengths; 400 nm cut-off filter for Width = 590 mm, depth = 650 mm, height = 367 mm; net weight 39 kg.
fluorescence measurements; three positions for user selected filters.
ORDERING INFORMATION

027.6200 CAMAG TLC SCANNER 4 I

for scanning by absorbance and fluorescence, equip-
ped for objects up to 200 × 200 mm, wavelength range
190 – 900 nm, complete with deuterium lamp, tungsten-
halogen lamp, and mercury vapor lamp, including Equilink
(027.6312) to winCATS, but without winCATS license.

winCATS Software and options:

027.6300 winCATS license including one year of internet
update service
027.6315 Quantitative chromatogram evaluation
027.6342 Spectrum library
027.6344 Track optimization
027.6346 Dual-wavelength scan
027.6348 Multi-wavelength scan
027.6340 Scanner selftest
027.6380 21 CFR Part 11 “compliance ready”

You can order the CAMAG TLC Scanner 4 as a complete

system:

027.6288 CAMAG TLC Scanner 4, complete system

027.6200 CAMAG TLC Scanner 4, for scanning by
absorbance and fluorescence, equipped for
objects up to 200 × 200 mm, wavelength
range 190 – 900 nm, complete with deu-
terium lamp, tungsten-halogen lamp, and
mercury vapor lamp
027.6312 Equilink to establish a 2-way communication
between the scanner and winCATS
027.6300 winCATS license including one year of
internet update service
027.6315 winCATS option “Quantitative chromato-
gram evaluation”
027.6340 winCATS option “Scanner selftest”

CAMAG TLC Scanner 4 18 | 19