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Understanding an HPLC system

In analytical chemistry, a High Performance Liquid Chromatography (HPLC) system is used for the purpose of separating, identifying, and the quantifying each of the various components that make up a mixture. A pressurized liquid solvent that contains the sample mixture uses pipes to pass through a column filled that is with a solid adsorbent material. An HPLC system is built in such a fashion that each component that goes into the sample has its own way of interacting with the adsorbent material. Each of these could be slightly different, as a result of which different components have different flow rates. As a result, components are separated in the process of flowing out of the column.

An HPLC is used in the following applications:

  • Legal, in which it helps law enforcement authorities to carry out critical tasks such as detecting the levels of banned substances in athletes' bodies
  • Medical, in which it is used for tasks such as knowing the content level of substances such as vitamin D
  • Manufacturing, where it is used in various production processes of many medical products
  • Pharmaceutical, biological and life sciences research

How is an HPLC system built?

Typically, an HPLC system consists of these core parts:

  • A sampler
  • A set of pumps
  • A detector

This is how these parts function in an HPLC system:

  • The sampler is used to deliver the sample mixture into the phase stream, which is mobile and is used to carry the mixture to the column;
  • The function of the pumps is to supply the mobile phase's required and set composition and flow through the column;
  • The detector is used to produce a signal that is relational to the quantity of the sample component that comes out of the column. This enables a quantitative analysis of the components of the sample.

The HPLC system is usually controlled by a user software and digital microprocessor, which assist in data analysis. A few types of mechanical pumps in an HPLC system are built to mix different kinds and samples of solvents together with their desired ratios made to change over time. This helps to generate a composition gradient right at the mobile phase. Most HPLC systems come with a column oven, which is used to adjust the temperature at which the separation is performed.



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