HPTLC-High performance thin layer chromatography

HPTLC introduction

HPTLC is an abbreviation for High-Performance Thin layer Chromatography or High-Pressure Thin Layer Chromatography.

This is a sophisticated advancement of Thin-Layer chromatography (TLC).

It has added advantages like better resolution, faster development of spots and also easy detection and quantification of separated compounds.

The advancement concerning better separation and resolution and faster resolution is due to

1) Use of ready-made HPTLC plates with optimized absorbent layers having smaller particles size with uniform particle size distribution as the stationary phase.

2) The use of pressure enables for faster development of chromatograms in even complex mixtures.

The HPTLC system is further automated to a greater extent than its precursor TLC as.

a) Auto sampling for application of samples on the plates avoiding manual sampling errors.

b) Online mixing of solvents for either gradient mode (i.e., a mixture of solvents in a predefined ratio) or isocratic mode ( single solvent) required during chromatogram development.High performance thin layer chromatography

c) Auto-detection of compounds from chromatogram using built-in UV and fluorescent detectors detector system.

d) Recording and storage of data from chromatograms utilizing an HPTCL data software in a computer.

Advantages of HPTLC over TLC:

♦ Samples in minute quantities like in nano-gram range can be detected using HPTLC.

♦ Handling and human errors are minimum due to automation.

♦ Better accuracy and sensitivity than TLC.


♣ The system is many folds expensive than TLC.

♣ Bulky instrumentation and large space requirement.

♣ Requires stringent condition of operation like dust free environment and temperature controlled conditions.

♣ A technically skilled person with the knowledge to run the system is needed.

Application of HPTLC:

1) For detection and analysis of components of phytochemistry, medicinal chemistry & organic chemistry.

2) In TLC one can identify the elements of an extract. Whereas using this method, one can even estimate it concentration.

2) Compounds having a complicated structure or those available in very scarce quantities can be analyzed.

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