Do you really know the High-pressure liquid chromatography?

(HPLC) is a type of chromatography in which the liquid transfer phase passes through a solid fixed phase column under high pressure. HPLC is a powerful analytical tool used to purify, separate, identify, and quantify each component of a mixture.

High-pressure liquid chromatography equipment consists of mobile phases, pumps, injectors, separation columns, and detectors. The sample material is analyzed by pumping the liquid phase (solvent) mixed with the sample into a column filled with solid-phase material. Each component of the mixture reacts differently to the absorbent material, separating the components of the sample material and letting it out of the column at different times. First of all, the material which binds well to the liquid phase comes out.
Next, the material that binds well to the solid phase comes out. High-pressure liquid chromatography is used to measure and purify the content of various components of the sample material. As an example, it can be used in the pharmaceutical industry. In this industry, HPLC is very useful for measuring the level of active drugs in certain materials.

Almost 90% of all High-pressure liquid chromatography systems use an UV-detector to measure the concentration through absorbance spectroscopy. The concentration is found through the Beer-Lambert’s Law: A = ε c L. The detector measures the absorbance versus time at one or more wavelengths. There are three types of detectors.

A single wavelength detector with a fixed optical filter, a selectable wavelength detector with a rotating grating and a multiple wavelength Diode-Array-Detector (DAD). Ibsen offer DAD spectrometers which are the fastest and most versatile form of spectrometer with UV-detectors.

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