High-Performance Thin Layer Chromatography (HPTLC)

High-Performance Thin Layer Chromatography

HPTLC is an automated form of Thin layer chromatography (TLC) which works on the principle of adsorption. It is used for qualitative and quantitative analysis. In this pre-coated plates with particle size of 5-6 um are used. There is a wide variety of stationary phases like silica gel is used for normal phase chromatography and C18 & C8 are used in reverse phase chromatography. This technique acquires the use of very less amount of sample with a very high accuracy of sample spotting with different apparatus. The sample spot used is less than 1 mm in diameter in this technique. Because of small and higher uniformity of size than TLC it has better efficiency.

The HPTLC instrumentation consists of sample applicators, a development chamber, scanner, and visualizer.

Derivatization can be done with the use of immersion device which leads to the quick and uniform spreading of reagent so that better detection can be there, auto reagent sprayer which provides fine and uniform spray. For spraying, there is new technology introduced called as micro-droplet technology, where spraying is done in a closed chamber with spraying nozzles.

Steps involved in HPTLC is shown in the figure below:

HPTLC has been advanced by its successful hyphenation with High-performance liquid chromatography (HPLC), Mass spectroscopy (MS) and with Fourier transform infrared spectroscopy (FT-IR) which helps in providing better analytical data of separated components.

 

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