HPLC | High performance liquid chromatography

HPLC is also known as high performance liquid chromatography or high pressure liquid chromatography. HPLC is usually a modified column chromatography. In HPLC, the a high pressure pump is attached with with the column. This high pressure pump can generate pressure upto 40Mp. The column is further filled with the adsorbent material that has very small particle size.
The small particle size gives a very large surface area for the sample molecules to interact. As a result the efficiency of separation increases giving high resolution.

Following are the different components of HPLC.
The column
The column is made up of stainless steel and can withstand very high pressure upto 50MP. The length of column can vary from 5 to 25cm and has an internal diameter of 4.5mm.

The stationary phase
The stationary phase is made uo of an adsorbent material that has very small particle size. The particle size is kept uniform to obtain better performance.

The mobile phase
Mixture of different solvents is used as a mobile phase. The solvents used depends on the type of sample molecules to be separated. The mobile phase solvents can be polar or non polar.

The detector
In order to detect the eluite (ie sample coming out of the column) the HPLC column is attached with the detector.

Видео HPLC | High performance liquid chromatography канала Quick Biochemistry Basics