A COMPARATIVE STUDY CONCERNING THE IMAGE ANALYSIS IN THIN LAYER CHROMATOGRAPHY OF FLUORESCENT COMPOUNDS

Abstract

The use of image analysis software for the detection and quantitative evaluation of fluorescent compounds in thin layer chromatography is investigated. Four different image analysis software packages (Sorbfil TLC, Biodit, JustTLC, and Melanie) were applied and compared. The image analysis methods were developed and validated for the simultaneous determination of quercetin and kaempferol in food supplements. The chromatographic separation used Silica gel 60 TLC plates as the stationary phase and carbon tetrachloride:acetone:formic acid (35:11:3, v/v/v) as the mobile phase. The plate images were recorded by a digital camera and imported in the image analysis software. The final results were validated by evaluating the linearity, precision, and accuracy of the methods used to quantify synthetic and real samples. All the results suggest that the image analysis software packages are proper to investigate the fluorescent compounds in quantitative thin layer chromatography.

Keywords:

• image analysis
• fluorescence
• kaempferol
• quantitative determination
• quercetin
• TLC

Notes

*SD, standard deviation.

**SEM, standard error of mean.

*Tablet weight: #1, 0.532 g; #2, 0.530 g; #3, 0.530 g.