HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY

Separates, identifies & quantifies molecules of differing polarities in the liquid phase

Sample Injection: A small amount of the sample mixture is injected into a column containing a stationary phase material.

01

02

Mobile Phase: A liquid solvent, called the mobile phase, is used to carry the sample through the column. The mobile phase can be a mixture of solvents with varying proportions to optimize separation.

Stationary Phase: The column is packed with a stationary phase material that interacts differently with various components of the

03

sample. This interaction leads to separation STOP

based on factors like polarity, size, and chemical affinity.

04 Separation Process: As the mobile phase passes through the column, the different components of the sample interact with the stationary phase to varying degrees. This interaction causes the components to elute out of the column at different times, creating distinct peaks in the chromatogram.

Detection: The eluted components are detected as they exit the column. Common detectors include UV-visible spectrophotometers, fluorescence detectors, and mass spectrometers.

06

05

Data Analysis: The resulting chromatogram, which shows the intensity of the detected components over time, can be analyzed to identify and quantify the different substances present in the sample.