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High Performance Liquid Chromatograph, HPLC, is a sophisticated instrument widely used in the food, pharmaceutical, and medical industries as well as in education and research.

Classical HPLC exploits the differences in the affinities of the components of a mixture for two different liquids to achieve the separation of the components and eventually their identification and quantification. One of the liquids is held on a solid support structure and is referred to as the stationary phase while the other liquid (usually a mixture of solvents) is allowed to move along the length and breadth of the solid support structure at a controlled flow rate.

Basically, an HPLC system consists of the following components:

  1. Bottles for holding and supplying the mobile phase solvents
  2. A pump that can deliver the mobile phase through the entire system at a determined and controlled flow rate. The pump should be programmable

3. A means of introducing the sample into the system; this could be manual or automatic

4. The column, which holds the stationary phase liquid and the solid support that holds the stationary phase. This is where the separation of components take place

5. A means of identifying and quantifying the separated components. This is called the detector

6. A means of processing and storing the results obtained. This is usually by a suitable software which also serves to automate the entire system

High Performance Liquid Chromatography is a well developed and highly robust technique and the HPLC instrument is basically easy to operate. However, there are recommended practices that should be followed to get reliable and high-quality results as well as preserve the life of the instrument.

These recommended practices are what we will be discussing in this series starting with basic housekeeping all through to how to get the best performance from the various components of the HPLC instrument.

So hang in there and I will see you next time with Best Practices on Housekeeping and how to handle solvents for HPLC

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Posted by Muyiwa Adebola

Muyiwa@aasnig.com