« FROM THE CHROMATOGRAM TO THE QUALITATIVE AND QUANTITATIVE RESULT »

Chromatogram Evaluation

Chromatogram evaluation with electronic image acquisition


For electronic image acquisition visible polychromatic light is captured. When long-wave UV light is used for object illumination, the camera captures the light emitted by fluorescent substances. When short-wave UV light is used for illumination of a chromatogram layer containing fluorescence indicator, the camera – like the human eye – registers the visible light emitted from the layer background. The quantification of image data is possible via the so called grey scale. Spectral selectivity is restricted to the colors of fluorescence. The strength of the electronic image acquisition is the overview of the complete chromatogram.

Chromatogram under white light
Chromatogram under white light
Chromatogram under UV 254 nm
Chromatogram under UV 254 nm
Chromatogram under UV 366 nm
Chromatogram under UV 366 nm

Chromatogram evaluation with classical densitometry

In classical densitometry the tracks of the chromatogram are scanned with monochromatic light in the form of a slit selectable in length and width. The spectral range of the CAMAG TLC Scanner 4 is 190–900 nm. Reflected light is measured either in the absorbance or in the fluorescence mode. From the acquired data quantitative results are computed with high precision and spectral selectivity.

With the TLC Scanner 4 absorption and fluorescence excitation spectra can be recorded. The strengths of classical densitometry as compared with image evaluation are spectral selectivity and the higher precision of quantitative determinations.

Recommendations
Applying samples in the form of narrow bands allows densitometric evaluation by aliquot scanning, i.e. scanning with a slit about 2/3 of the track width. This improves reproducibility as the center portion of the sample zone is homogeneous and positioning errors, which can occur with samples applied as spots, are avoided.

For quantification sample zones should always be scanned with the wavelength of maximum absorbance which can be determined by spectra recording or by multi-wavelength scanning.

Products

BioLuminizer®
The CAMAG BioLuminizer® is a detection system specifically designed to detect bioluminescence on HPTLC plates.
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TLC-MS Interface 2
The elution-based CAMAG TLC-MS Interface 2 is a highly convenient and versatile instrument allowing for rapid and contamination-free elution of TLC/HPTLC zones with online transfer to a mass spectrometer.
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TLC Visualizer 2
Professional imaging and documentation system for TLC/HPTLC chromatograms with a new digital CCD camera, connected by USB 3.0
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TLC Scanner 4
The CAMAG TLC Scanner 4 is the most advanced workstation for densitometric evaluation of TLC/HPTLC chromatograms and other planar objects.
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UV Lamp 4
The CAMAG UV Lamp 4 is designed for use in a TLC laboratory.
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UV Cabinet 4
The UV Cabinet 4 is designed for inspecting thin-layer chromatograms or other objects under UV light in absence of ambient light.
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