winCATS – a new tool for Planar Chromatography

winCATS - Planar Chromatography Manager is a completely new approach to modern Thin-Layer Chromatography. The winCATS Software is based on a variety of modules to manage, monitor, and control all steps of the planar chromatographic procedure from sample application, to TLC plate development, classical densitometry and image documentation.

winCATS is designed to manage and supervise all software-driven CAMAG instruments. For non-software driven equipment or chromatography tasks e.g. sample application via Nanomat and pre- or post chromatographic derivatization, the user can manually enter all related parameters and winCATS will archive and report them in compliance with GMP/GLP.

April 2003: winCATS - Planar Chromatography Manager is now highly appreciated by our customers as a user friendly software. This shows that our approach to monitor all steps of the Planar Chromatographic procedure from one single software was right.

Currently available are EquiLinks for the following CAMAG instruments and winCATS Software options:

• Linomat 5
• Automatic TLC Sampler III
• Automatic TLC Sampler 4
• Automated Multiple Development 2
• TLC Scanner 3
• Digital camera (Canon Power Shot G2)
• Video camera HV-C20
• Option Quantitative Evaluation
• Option Scanner 3 Qualification
• Option Scanner 3 Dual Wavelength scan
• Option Scanner 3 Track Optimization
• Option Spectrum Library
• Option ATS4 FreeMode

winCATS basic version features:

• Input of TLC plate specific data
• Input of the sample(s) and substance(s) data
• Manual entry of the application sequence for manual sample application devices, such as the CAMAG Nanomat 4 or the CAMAG Linomat IV
• Manual entry of the developing parameters for manual / semi automatic developing devices, such as Twin Trough Chamber, Automatic Developing Chamber, and AMD
• Manual entry of pre- and post-chromatographic derivatization parameters
• Documentation step: Import of VideoStore images into the analysis file
• Generate a customer configurable report in conformity with GxP
• Instrument properties offering information about the connected instruments
• Extensive context sensitive HELP functions and an interactive tutorial
• If a TLC Scanner 3 is available, the basic version also supports scanning, spectra recording, baseline correction, peak detection, manual peak editing, spectra display and export of peak data to i.e. Excel.

The winCATS Planar Chromatography Manager organizes your workflow in the following steps: definition of plate, samples and standards, sample application, development, derivatization, detection, evaluation, and documentation.

Workflow of Planar Chromatography:

Sample preparation

Planar Chromatography does not require the same sample preparation effort as other chromatographic techniques. Simple sample filtering is, in most cases, all that is needed because the plate is used only once.
winCATS does not incorporate the sample preparation step, but the TLC/HPTLC user knows the importance of sample preparation and that his analytical results depend on the preparation of samples & standards.

The TLC step <Stationary phase> and <Definitions>

All required components necessary to run an analytical task are usually defined in the appropriate SOP but the user still needs to enter all observations and details into his lab journal so that such information can be referenced together with the results of the analysis. winCATS offers pre-defined dialogs, selections and free text input fields designed for completely documenting all these details during TLC/HPTLC analysis.

Important information, like plate type, plate manufacturer, plate size, solvents, pre-washing, etc. can be entered in the 'Stationary Phase' dialog window of winCATS.

Stationary plate dialog window

Definitions – Screening

The <Definition - Screening> dialog covers all sample related parameters, such as sample name(s), substance name(s), Rf of the substance(s), manufacturer, expiry dates, etc.

Substances definitions in "screening" mode

Sample definitions in "screening" mode

The TLC step <Sample Application>

Sample application is the most crucial step in Planar Chromatography. Errors that occur during sample application cannot be compensated for in chromatogram development or detection and evaluation.
CAMAG offers a range of devices for good sample application. The range of applicators starts with a simple low cost hand operated Nanomat 4 for spotwise application, a semi-automatic bandwise applicator like the new Linomat 5 and the fully automated Automatic TLC Sampler 4.

Sample application sequence table for the Nanomat 4

Sample Application by Nanomat 4

The Nanomat 4 is a hand operated application device, which cannot be controlled by winCATS. However, to comply with GMP/GLP, all application parameters for the Nanomat 4 can be entered, thus completing the analysis report.
The screenshot shows the sequence table for spotwise sample application with the Nanomat 4 in combination with Capillary Dispenser. Due to the size of the glass capillaries, only fixed volumes 0.5, 1.0, 2.0 and 5.0 µL can be selected.

Sample application sequence table for the Nanomat 4

Sample Application by Linomat IV

The high number of Linomat's in use worldwide shows, that the Linomat band applicator is the workhorse in many laboratories. Although the Linomat IV cannot be controlled by winCATS, the application parameters entered are used for calculation of results.
The screenshot shows the sequence table for sample application with the Linomat IV. In this table the user enters all parameters necessary for applying the sample: application volumes, the sample vials, application positions on the TLC plate, dosage speed, etc.

Sample application sequence table for the Linomat IV

Sample Application by Linomat 5

The new Linomat 5 is the successor of the Linomat IV and can be used as a stand-alone unit or controlled by winCATS. The semi automatic functionality of all Linomat's remains, i.e. the user fills the syringe manually and inserts it into the Linomat 5 turret. The Linomat 5 can be managed and controlled from winCATS were the dosing parameters and application sequence can be entered.

Sample Application by Automatic TLC Sampler III

Although this instrument is no longer in production, we have still a large number of satisfied ATS3 users. This fully automatic applicator can be connected to winCATS via an EquiLink. The dosage parameters and application sequence can be entered in winCATS, which will then monitor the sample application.

Sample application sequence table for the Automatic TLC Sampler III

Sample Application by Automatic TLC Sampler 4

The Automatic TLC Sampler 4 is the successor model of the ATS3 and is always controlled by winCATS. The dosage and rinsing parameters, the application sequence for up to 66 vials, application positions on the TLC plate, application type (spotwise or bandwise), dosage speed, etc. are entered in winCATS, which then monitors the progress during sample application.

Sample application sequence table for Automatic TLC Sampler 4

The TLC step < Development >

Presently winCATS is unable to monitor the development step. The parameters for the development devices Twin Trough Chamber, Horizontal Developing Chamber and Automatic Development Chamber (ADC) can be entered manually into winCATS and are saved together with the analysis. The screenshot shows the parameter of Twin Trough Chamber.

Developing parameters using a 20 x 10 cm twin trough tank

Development by Automatic Developing Chamber (ADC)

Many ADC users asked us to implement the ADC into winCATS. It has always been CAMAG's intention to implement full control of the ADC but the concept of communication in winCATS does not allow full external control of this device. To fulfill our customers request as well as possible, we implemented an ADC specific dialog.

winCATS developing parameters for ADC

Development by Automated Multiple Development (AMD)

Although the Automated Multi Developing chamber (AMD2) is not yet implemented in winCATS, it is now possible to enter the AMD developing parameters. All relevant parameters, like solvents, drying time, etc. can be entered.

winCATS developing parameters for AMD

Development by Automated Multiple Development (AMD2)

The parameters for the gradient development with the Automated Multiple Development chamber (AMD2) can be entered into winCATS and are saved together with the analysis. The screenshots are showing parameters like solvents, drying time, etc. and the gradient for the AMD2

winCATS developing parameters for AMD2

Gradient editor

Gradient in graphic format

The TLC step < Pre- / Post Chromatographic Derivatization >

None of the derivatization devices or tools can be controlled by winCATS. The derivatization parameters, like reagent, treatment type (spraying or plate immersion), plate heater, temperature and time are entered manually into winCATS to make the documentation complete

winCATS derivatization parameters

The TLC step < Detection >

For densitometric evaluation of a TLC/HPTLC chromatogram, the light reflected by the plate is measured by means of a photo sensor. Densitometric measurements can be performed in absorbance or in fluorescence mode.

Detection by means of the TLC Scanner 3

The CAMAG TLC Scanner 3 measures the diffusely reflected light from the plate, illuminated by a light beam with selectable length and width. A photo sensor transforms the light into current, which can be further processed in winCATS.
All useful parameters from previous TLC steps, such as positions from sample application and chromatogram development, can be automatically used to define the scan pattern. If the correct wavelength and slit size has been set, the scanner can start scanning the TLC plate.
winCATS permits scanning the plate using one single wavelength, two wavelengths (for subtraction) and in multi wavelength scan mode. The possible wavelength range is 190~800 nm with automatic lamp changing in case the wavelength passes the boundaries of a lamp.
Spectra from all detected peaks can be recorded automatically and used for peak identity or purity comparisons.

Scanner 3, 1WL dialog for selection of lamp, slit, wavelength, filter, scanning mode, etc.

Peak display after Integration

Automatically recorded Spectra of the integrated peaks

Option: Dual Wavelength Scan (027.6346)

With some samples the "Dual Wavelength" scan mode can achieve a better result because it will subtract the data of the 2nd WL from those of the 1st WL. The plate is scanned twice with different wavelengths to achieve this. Data from the first and second scan are both shown in the background correction view. The program automatically calculates the difference between the two data curves and draws the new analog curve.

Scanner 3 – Scan parameter for Dual wavelength

Integration after Dual-wavelength scan. The blue curve was measured at 254nm and the red curve at 450 nm

Option: Multi Wavelength Scan

Multi Wavelength Scan allows scanning of the entire plate with up to 31 different wavelengths, freely selectable between 190~800nm. The user can select between a) fixed increments, b) free selectable wavelengths or c) wavelengths as they are individually assigned to each substance in the substance table.

Scanner 3 - MWL Detection

3D view of one track at all wavelengths

3D view of all tracks at one wavelength

After the MWL scan winCATS will / can evaluate / calculate the results for each substance using the peak data generated at the wavelength assigned to that substance. Result calculation using the optimal wavelength for each individual peak improves the accuracy and is a new feature in winCATS. No other TLC evaluation software package available today offers this advantage.

Substance assignment in MWL mode

Option: Track optimization (027.6344)

By means of this option distorted chromatograms can be measured and evaluated. Each track is measured several times at slightly different x-positions and the software calculates the optimized path of the measurement.

Trace of a track after optimization. The black line shows the path of the final analog data used for this track.

Option: Scanner Qualification (027.6340)

The software option "Scanner qualification" monitors all mechanical, electronic and optical parts of the CAMAG TLC Scanner 3. Automatic re-adjustment of lamps and grating can be performed and will bring performance back to factory specifications, which may save costs for service calls.
Note: The Scanner Qualification option 027.6340 is a prerequisite if IQ-OQ is to be performed on the specified TLC Scanner 3.

Screenshot of the TLC Scanner 3 qualification report

The TLC step < Evaluation >

During quantitative evaluation, the difference between the optical signal from the sample free background and that from a sample zone (fraction) is correlated with the amount of the respective fractions of calibration standards chromatographed on the same plate.

Option: Quantitative Evaluation (027.6315)

The option 027.6315 "Quantitative Evaluation" permits calibration of up to 100 substances against their standards chromatographed on the same plate. Several different calculation routines, like Reproducibility, Single- or Multi-Level calibration are available.
With Single-Level calibration a single standard level, preferably applied in duplicate or triplicate, is used for reference. The peak height and/or area of the unknowns are calculated against this calibration standard. winCATS provides, besides the results of the unknowns, the correlation factor of the standard curve (if possible), the sdv% (residual standard deviation between standard points and curve) and CV% (coefficient of variance between replica of one sample).

Multi-Level calibration can use of up to 10 different standard levels (again if possible in duplicate or triplicate). The program calculates for each substance the selected calibration curve. All results from calibration and for unknowns are reported as with single level calibration.
Besides the common linear- and polynomial regression routines winCATS offers two Michaelis-Menten regression modes. Michaelis-Menten 1 is always running through the origin and thus the only non linear curve available with only two standard levels, while Michaelis-Menten 2 is an optimized version, needs three standard levels and does not necessarily run through the origin.
In winCATS you can use absolute amounts for each substance or, in the stock solution mode, you can enter the amounts as they appear on the balance. In stock solution mode you also enter the solvent this amount was dissolved in, dilution volume(s) or factor(s) etc. winCATS will use all these parameters to calculate the results.

Absolute mode

Stock solution mode
For the unknown samples you can enter the dilution and a target weight (e.g. average tablet weight) and winCATS will calculate the result relative to this target weight. The software also permits the calculation of sub-components (related compounds).

Multi level calibration curve with 5 standard levels.

Calculation of % active component in a sample can be calculated by referring the result to the reference amount of the sample (Sample reference amount mode only).

If the option Quantitative Evaluation is available you can also calculate Identity and Purity of peaks if the respective spectra have been measured.

Peak-Identity is calculated by correlating the spectra of the unknown peaks to the spectra of co-chromatographed standards. Peak-Identity graphic display. Spectrum of an unknown peak is overlaid with the spectra of the known standard(s). A correlation graphic can be shown for evidence (see below).

Peak-Purity is calculated by correlating the spectra measured within a peak. In this case spectra are automatically measured in both the peak-flanks and at peak-maximum. Peak-Purity correlation display. Spectra measured at 3 positions within an unknown peak are overlaid for "purity" for comparison. (see above).

Option: Spectrum Library (027.6344)

The spectrum library option permits to save measured spectra into a library, which can then be used to identify unknown substances on other plates. Several search criteria are built-in and enables you to find the best hit. The screenshot shows the unknown substance spectra from the TLC plate on the left, while all spectra from the library matching the search criteria are shown on the right. The hit list is shows 5 matching spectra with the best match in first position - here codeine.

Spectrum Library Spectrum of an unknown peak is overlaid with a spectrum from the library

The TLC step < Documentation >

One of the merits of Planar Chromatography is the possibility to take a picture of the result of the analysis. A vivid image of a TLC plate gives more information then 1000 words in writing.
In the early days of documentation ordinary photography was widely used and was slowly replaced around 1975 by Polaroid instant photography. In the early nineties CAMAG introduced TLC documentation based on Video Technology using a video camera. Equipment for digital photography became more and more affordable and in the year 2000 CAMAG introduced the Reprostar 3 with Digital camera for TLC documentation.

Option: Documentation with Digital camera (027.6370)

winCATS is able to control all functions like aperture, zoom, exposure time, sensitivity, etc. of the digital CANON PowerShot camera. This means you can take a picture of the TLC plate using predefined settings and the captured image is automatically transferred to winCATS and becomes part of the analysis, in compliance with GMP/GLP.
Exporting the image to VideoScan or a 3rd party software e.g. Word, is possible

Camera control box with image preview and all camera settings - Zoom, Exposure time, Aperture, etc.

Option: Documentation with Video camera (027.6375)

winCATS is able to control all functions of the HV-C20 video camera. This means you can capture an image of the TLC plate with the HV-C20 video camera and it becomes a part of the analysis, in compliance with GMP/GLP.

Camera dialog with image preview.

The captured image can also be exported to a .cpf file and evaluated in VideoScan or exported to clipboard for pasting into 3rd party software e.g. Word or PowerPoint.
Existing VideoStore 2 users can order the EquiLink 027.6375 from CAMAG and use their HV-C20 Video camera from the winCATS platform.

Image import

winCATS allows import of images captured or modified by other 3^rd party software but all these images are automatically supplied with a GMP/GLP warning, which is always shown and printed with the image properties.

Exceptions from this rule are the images captured with the CAMAG VideoStore system as these images comply with GxP.

An image captured by VideoStore and imported into winCATS

Image annotation

winCATS can annotate the images, no matter if these are captured or imported. Any horizontal or vertical text, boxes, circles, freehand objects etc. can be added to the image.
The application position and solvent front lines can be set and allows display of Rf value for any position of the plate.

Annotated image in winCATS

Showing Rf value

Report

No job is finished without the paperwork. This means only the final printed report shows the result of the analysis. What should be reported? The report setup window shows a copy of the TreeView, the items to be printed can be selected individually. The report shows the logged in operator's name, date & time, page number & total pages and an individual report ID number automatically generated by the system.

Report setup, a copy of the TreeView is shown in the Report Setup window and allows selection of items to be printed

Save to disk

winCATS uses two file types, the method file .cme and the analysis file .cna. The method [.cme] contains all parameters to perform the analytical task - in accordance with the SOP, while the analysis file [.cna] contains the parameters actually used AND all the data acquired during the actual analysis.

Option: 21 CFR part 11 'compliance ready' (027.6390)

CAMAG has implemented this FDA requirement in version 1.3.0 released in Nov. 2003. For details please check our standardized answer to vendor checklists.

Additional features, tools, information:

New 3D graphic tool
A 3D graphic tool showing all tracks has been implemented. The angle of view can be rotated at will by simultaneously holding the CTRL-key and moving the mouse with pressed left mouse button.
Additional display features, that can be accessed via a right mouse click and selection from the context menu, are: export to clipboard, export as a .bmp data file, print the graphic, data point representation, changing the order of view e.g. track 1 in the front or track 1 as last displayed track, display the migration normal from left to right or reversed from right to left, etc..

Export to clipboard, saved as BMP file or send directly to a printer

A winCATS 3D-grapic copied into a Word document.

New 3D-View with "tool tip"

This "tool-tip" shows the actual signal strength [AU], X-position on the plate [mm] and migration distance [mm or Rf]. In the MWL mode, it will display in addition the actual wavelength [nm]. Just click on one of the analog curves to see information of this position. With the left / right cursor keys you can move along the selected curve. Jump from track to track with the up / down cursor keys

The "tool-tip" shows important values for each position of the analog curve

User login and user rights

In winCATS exists the possibility to enter new users and assign certain user rights including a password. The possible user privileges are: administrator, method editor and method user. The administrator can assign/delete users, install/remove equipment and software options. The method editor can create and execute any method and analysis and the method user can call up an existing method and start his analysis according the parameters given in the method.

Assignment of a new user with his own password.

Extensive context sensitive HELP

Included in winCATS is a comprehensive HELP, which should be able to answer all your winCATS and some TLC related questions. Wherever you need HELP in the program - press the <F1> key to get context sensitive HELP on this part of the program.

Also included in the HELP are all the manuals of the different CAMAG instruments, a glossary, theoretical aspects around modern TLC, mathematical formulas, error messages, etc. in other words a wealth of information.
With the implementation of new features, also the winCATS HELP has been extended. HELP required for entering the Linomat IV parameters, MWL evaluation, Main- and Sub-component calculation are called-up by just pressing the F1 button.

winCATS Context sensitive "HELP"

Easy software update
During the warranty period of 12 months all winCATS users qualify for free update to the latest winCATS version by downloading the necessary files from the Internet. After 12 months, this software update service can be renewed at moderate cost.

Upgrade of existing CAMAG instruments for winCATS
Upgrade packages are available for existing CAMAG TLC Scanner 3 with DOS based CATS software and for CAMAG Automatic TLC Sampler III with older ATS3 version 2.xx
CAMAG offers upgrade prices for (DOS) CATS software options purchased at list price to the corresponding winCATS successor products.

winCATS Demo CD
A winCATS Demo CD-ROM is available free-of-charge. Instruments cannot be controlled by the Demo-Version, but all the features of this new software can be tried out. The HELP system and the interactive tutorial are fully functional.
Do you have an older Demo version of winCATS? You can download the current version from the CAMAG Internet web page and update your existing Demo version at no charge.

OFFLINE license and multiple licenses

Besides ONLINE licenses CAMAG can now also offer so-called OFFLINE Licenses.
In an ONLINE mode, there is a direct communication between winCATS and the device. The controlling PC with winCATS is mostly located in a direct neighborhood of the instrument(s) and takes over the control of that particular device. For this online mode winCATS together with the respective EquiLink is required. Data can be saved via a Network on to a server.
In the OFFLINE mode, the user can access the data from his office via the network. Now he can prepare the methods or perform re-calculation of the analyses from a PC not directly linked to the device. Methods, which have been prepared 'offline' can be send via the Network to the 'online' workstation. Analyses of an 'online' station can be evaluated on a PC with appropriate 'Offline' licenses.
Multiple licenses: CAMAG can provide multiple winCATS licenses with quantity discount for the same customers.
Contact CAMAG and request a quotation for your OFFLINE and/or multiple licenses. Inform us which instruments and which software options you need to use and we will provide you a tailor made quotation.

Service and support
In case you need assistance, call your local distributor or contact info@camag.com
Further information on winCATS is available on www.camag.com

Training
Training courses for customers and prospects are offered by CAMAG Muttenz / Switzerland, by CAMAG USA / Wilmington NC, by Anchrom / Mumbai India and in Chinese language by CCSU in Guangzhou or Shanghai. Also in-house courses at customer sites can be arranged. Request details by fax or visit our web site www.camag.com