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The main focus of this chapter is the theoretical and instrumental processes that underpin densitometric methods widely used in thin-layer chromatography (TLC). Densitometric methods include UV–vis, luminescence and fluorescence optical measurements as well as infrared and Raman spectroscopic measurements. The chapter is divided in two general parts: a theoretical part and a practical part. The systems for direct radioactivity measurements and the combination of TLC with mass spectrometry are also discussed. All these systems allow measuring an intensity distribution directly on a TLC plate. We call this “in situ detection” because no analyte is removed from the plate.
The main focus of this chapter is the theoretical and instrumental processes that underpin densitometric methods widely used in thin-layer chromatography (TLC). Densitometric methods include UV–vis, luminescence, and fluorescence optical measurements as well as infrared and Raman spectroscopic measurements. The chapter is divided in two general parts: a theoretical part and a practical part. The systems for direct radioactivity measurements and the combination of TLC with mass spectrometry are also discussed. All these systems allow measuring an intensity distribution directly on a TLC plate. We call this “in situ detection” because no analyte is removed from the plate.
Thin-layer chromatography (TLC) is a well-established and widely used separation technique. Most undergraduate students of chemistry or food science used TLC as a primitive separation tool, which does not need more than small pieces of TLC plates, a glass jar and some solvents. TLC has evolved from a simple separation method of the past into an instrumental technique that offers automation, reproducibility and accurate quantification for a wide variety of applications [1]. The use of modern 10*10 cm TLC plates with narrow particle size distribution is called high performance thin layer chromatography (HPTLC), to distinguish the method from the use of traditional 20 20 cm TLC plates.