Chromatography in chemistry is the science of separating
mixtures. To do this, the sample goes through two distinct phases, named the
mobile phase and the stationary phase. In which, the stationary phase remains
fixed, while the mobile phase flows from the system. The mobile phase is the
liquid or gas that runs through the system separates the molecules in the
stationary phase at different rates. The stationary phase is either the solid
or liquid (e.g. Column, silica, paper, TLC plate) at which molecules are to be
separated.
The mobile phase is an important parameter in the
chromatographic separation process, and many of its other properties are
significant in HPLC analysis. The mobile phase is a developing liquid or solution
combination that travels up to the stationary phase, moving samples with it.
The analytes of the sample solution separate, based on how much they adsorb
onto the stationary phase versus how easily they dissolve in the system of the
mobile phase. This is an important role of the mobile phase in HPLC and other
chromatography. The retention time of each molecule will contrast, it depends
on the interaction between the stationary phase, molecules, and solvents are
being used in the analysis. As the sample travels through the HPLC column, it
interacts at different rates between the two phases, mainly due to different
polarities in the molecules. Generally in RP-HPLC consists of a mixture of
methanol, acetonitrile, tetrahydrofuran (THF), propanol, and water sometimes
buffer (e.g. Acetate buffer, phosphate buffer) used as a mobile phase.
The mobile phase of HPLC should have the following
important features to meet the essential objectives:
- The sample to be analyzed has to be soluble in the mobile phase.
- The components of the solvent system have to be non-toxic and non-hazardous.
- The mobile phase must be inert; they should not react with the sample components and the stationary phase.
- When it passes through the detector it should not give the response.
- The mobile phase should be inexpensive and make the proportion of solvents used cost-effectively viable.
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