Before we start the discussion about HPLC, we need to understand what kind of problem we can solve with HPLC.

Problem

In column 1 there are four components, component A, B, C, and D. If all the materials are present separately it is easy to determine the quantity of each material by simply weighing. However, when all the components are present as a mixture, separation and quantification by physical methods become challenging.

Technique

Here, advanced analytical techniques like HPLC come into the picture. In this technique, components are separated, detected, and quantitated by HPLC.

Solution

After the sample injection, a chromatogram is obtained which gives the result. In the chromatogram, retention time serves as a qualitative parameter, indicating which compound is present in the sample. Another parameter, peak area/peak height, serves as a quantitative parameter and provides information about how much quantity of compound is present.