High-Pressure Liquid Chromatography (HPLC)

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High-Pressure Liquid Chromatography (HPLC) is a separation technique in which a liquid or properly dissolved solid sample is passed through a column at high pressure.

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It is also referred to as High-Performance Liquid Chromatography

The basic principle of HPLC is that it separates a sample into its constituent parts based on the relative affinities of distinct molecules for the mobile phase and the stationary phase used in the separation.

The molecules are retarded while passing through the stationary phase, depending on the chemical structure of the analyte.

High Performance Liquid Chromatography Instrumentation

1. Solvent reservoir 2. Pump 3. Injector 4. HPLC column 5. Detector 6. Computer

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Application of HPLC

– Pharmacetical applications for the analysis of drugs. – Purification of water – Separation and analysis of non-volatile or thermally unstable compounds. – Ion-exchange Chromatography of proteins

Advantages of High Performance Liquid Chromatography

– High resolution – Quick analysis – Separation of volatile and non-volatile components

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Disadvantages

– High cost – Relatively difficult to operate

FAQs

What is the difference between high pressure and low-pressure liquid chromatography?

There’s no relative comparison between high pressure and low-pressure liquid chromatography. The term “high” in high-pressure liquid chromatography is used as the high pressure is required to pass the mobile phase through a column packed with the stationary phase.

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