Sunday, 30 September 2012

What Is HPLC? High performance liquid chromatography.


Brief explanation about what is HPLC

HPLC (High-performance liquid chromatography) is also referred as High-pressure liquid chromatography.

It is a technique (chromatographic) used for the separation of a mixture of various compounds in biochemistry as well as in analytical chemistry. In this process the separation technique involves the injection of very small volume of a sample to be separated into a tube which is packed with a tiny particles (particle size in 3 to 5 micron in diameter) which is known as stationary phase. Now the mixture to be separated  or the unknown sample is passed or moved down the packed tube known as column, with the help of a liquid known as mobile phase, is forced down through the column by providing high pressure and then delivered by a pump.

These components are then separated (based on the polarity of the compounds) from each other with the help of  column packing which involves various physical and/or chemical  interactions between their molecules and the packing particles. Now the separated components are detected by the detector attached at the exit of the tube or column. The output or the signal which is detected by the detector is known as chromatogram and the entire technique is known as chromatography and the instrument which is used to separate unknown mixture is known as High performance liquid chromatography (HPLC).

To know "What is HPLC" it is necessary to know the components of HPLC:

  1. Pump
  2. Injector
  3. Column
  4. Detector
  5. Recorder
  6. Degasser
  7. Column Heater

Types Of HPLC is must to understand What is HPLC:

  1. Partition Chromatography
  2. Reverse phase Chromatography
  3. Normal phase Chromatography
  4. Displacement Chromatography
  5. Size-exclusion Chromatography
  6. Ion exchange Chromatography
  7. Affinity Chromatography

 I hope you might have got some of the idea regarding "What is HPLC", the basic principle of HPLC , components as well as types of HPLC.

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