1. Hyphenation of planar chromatography with chemometrics Petar Ristivojević, Dušanka Milojković-Opsenica, Gertrud Morlock

2. Column chromatography with different detections Planar chromatography
1. Convert to numerical data
2. Image analysis
Multivariate analysis
Multivariate analysis

3. Signal preprocessing in HPTLC research
Scaling and centering
Auto-scaling
Mean centering
Warping image procedure
Correlation optimized warping (COW)
Dynamic time warping
(DTW)
RGB
Normalization
Standard normal variate (SNV) R G B

4. Principal Component Analysis
PCs score plots of German propolis according to the red channel.

5. Image analysis procedures
PCA based on peak area of characteristic bands for blue channel: A) Image analysis B1) score plot, B2) loading plot for PC1, B3) loading plot for PC2.
PCA based on intensities of pixels along the solvent front for red, green and blue channels: A) Image analysis B1, B4, B7) score plots for red, green and blue channels, B2, B5, B8) loading plot for PC1, B3, B6, B9) loading plot for PC2.
PCA based on mean value of selected zone for gray scale: A) Image analysis, B1, B4) score plot; B2, B5) loading plot for PC1, B3, B6) loading plot for PC2.

6. Phenolic profile of German propolis
HPTLC fingerprint of German propolis

7. Pattern recognition of German propolis
Clustering on a 3D PC-score based on zone intensities of the HPTLC image

8. Prediction of biological activity using HPTLC and chemometrics?

9. Statistical parameters for the six PLS models
Bioautography assay of propolis samples against six bacterial strains. A) phenolic profile of propolis extract, B) E. faecalis,C) B. subtilis,D) S. aureus,E) L. monocytogenes,F) A. hydrophila andG) S. flexneri.
Regression coefficients of PLS models obtained from six bacterial strains.

10. Pattern recognition of German beer samples
The HPTLC phenolic profile of German beer sample: A) without derivatization, B) after derivatization with Neus reagent and PEG, C) after neutralization with phosphate/citrate buffer, D) after DPPH assay, E) after Aliivibrio fischeri assay, F) after B.subtilis assay, G) after AChE assay.

11. PCA 3D score plot based on: A) DPPH assay, B) A. fisheri ssay, C) B
PCA 3D score plot based on: A) DPPH assay, B) A. fisheri ssay, C) B. subtilis assay, D) AChE assay.

12. RGB channels (A) and HPTLC chromatograms of the phenolic profiles of the blue-type (B) and orange-type (C) German propolis samples.
Multivariate analysis using rTLC software

13. Thank you for your attention!
German Academic Exchange Service (DAAD)
Ministry of Education, Science and Technological Development of Serbia, Grant No