Description
High performance liquid chromatography (HPLC)-Introduction, theory, instrumentation, advantages and applications
The obvious way to increase the flow rate and get efficient separation is to force the liquid by a positive displacement pump or by gas pressure. This versatili;y can be achieved by
making certain modifications in columns and by using smaller diameter and smaller surface area of column particles and by using other suitable packing structure. This is HPLC i.e. High Pressure/Performance Liquid Chromatography.
Thus, HPLC is a high resolution and high speed liquid chromatography. It has Several times more resolving power than open column liquid chromatography. Hence it is used for speedy resolution of complex mixtures.
ADVATvTAGES OF HPLC
1. It provides a specific, sensitive and precise method for analysis of different
complicated samples.
2. There is ease of sample preparation and sample intrtduction.
3. There is speed of analysis.
4. The analysis by HPLC is specific, accurate and precise.
5. It offers advantage over gas chromatography in analysis of many polar, ionic
substances, metabolic products and thermolabile as well non-volatile substances.
INSTRUMENTATION OF HPLC
A line diagram of HPLC unit is shown in
In order to attain reasonable high flow rates and yet keep particle size ofpacking very low (3-10 pm), pumping pressures of several hundred atmospheres (2000 – 8000 psi) are required. Thus, the equipment for HPLC is quite elaborate though simple.
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