Principle & working of HPLC

Question

Accepted Answer

HPLC is a technique used to separate, identify, and quantify components in a mixture based on their interactions with a stationary phase and a mobile phase.

HPLC stands for High-Performance Liquid Chromatography.
It involves the use of a stationary phase (usually a column packed with particles) and a mobile phase (liquid solvent).
The sample mixture is injected into the HPLC system and carried by the mobile phase through the column.
Different components in the mixture interact differently with the stationary phase, leading to separation.
Retention time is the time it takes for a component to elute from the column and is used for identification.
Detection can be done using various techniques such as UV-Vis spectroscopy or mass spectrometry.
HPLC is widely used in pharmaceutical, environmental, and forensic analysis.
Example: HPLC can be used to analyze the concentration of drugs in blood samples.

Accepted Answer

The separation principle of HPLC is based on the distribution of the analyte (sample) between a mobile phase (eluent) and a stationary phase (packing material of the column). Depending on the chemical structure of the analyte, the molecules are retarded while passing the stationary phase.

Principle & working of HPLC

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Principle & working of HPLC