What is basic principle of hplc?

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The separation behind HPLC is based on the polarity of the compounds.
{Polarity refers to : the greater the difference in electron affinity i.e. electronegativity between atoms in a covalent bond, the more polar the bond.partial negative charges are found on the most electronegative atoms, the others are partially positive.The moleacular electrostatic potential energy of a hydrogen ion at a particular location near a molecule. Negative electrostatic potential corresponds to : partial negative charges , Positive electrostatic potential corresponds to : partial positive charges.} It is used to analyze,identify,purify and quantify componds. It has a mobile phase,a stationary phase,detector.The mobile phase is continuously pumped at a fixed flowrate through the system and mixed by the pump.The injector is used to introduce a plug of a sample into the mobile phase without having to stop the mobile phase flow,& without introducing air into the system. The mixture of components is carried in a narrow band to the top of the column.Some compounds in the sample mixture will have greater preference for stationary phase than the mobile phase and will be retained in the column longer. POINT TO REMRMBER : "Like attracts Like" i.e if the column is non-polar the compound to elute first will be the most polar one. Those components that are not retained as strongly as are carried by the mobile phase down the column. The detector is then used to respond to a physico-chemical property of analyte,this response is digitally amplified and sent to a data system where it is recorded as a 'chromatogram'.


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