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What is the principal of HPLC?

Updated: 8/10/2023
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11y ago

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HPLC - High Performance Liquid Chromatography

Principle of hplc -

HPLC seperates mixture of compounds on the basis of polarity.

{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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9y ago

High performance liquid chromatography (HPLC) is used to separated components in a mixture, identify them, and quantify them. Pumps push pressurized liquid solvent w/ mixture though a column with adsorbent. Each component will interact with adsorbent differently and have different flow rates, causing separation.

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14y ago

The separation behind HPLC is based on the polarity of the compounds. "Like attracts Like" i.e if the column is non-polar the compound to elute first will be the most polar one.

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11y ago

as the eqbm. attained in between mobile and stationary phase pigments get adsorbed on column and seprated

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HPLC means Hipressure liqied chromatogaraphy &Hiperformence liqued chromatography

PRINCIPAL:the main principal of HPLC is adsorbtion the princepal of separation is highly

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NP-HPLC is "Normal Phase" HPLC, wherein the solvents used are less polar than the substrate in the HPLC column (e.g. using hexane or dichloromethane with a silica HPLC column). RP-HPLC is "Reverse-Phase" HPLC, wherein the solvents used are more polar than the substrate in the HPLC column (e.g. using Water and Methanol with a octadecylsilane (ODS or C18) column).


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