HPLC Definition , Principle , Advantage and Disadvantages :

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Definition

Molecules seperate with terrible pressure High performance liguid chromatography. High performance liguid chromatography due to working with high pressure. The polarity base is the separation of molecules.

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Hplc
Hplc

Components : Normal Phase And Reverse Phase

Analyte Soluble sample Soluble sample
Stationary Phase Polar Hydrophilic Liquid / Solid

( Silica )

Non Polar Hydrophobic Ligid / Solid.

( Silica Coasted with long Chains of hydrocarbons ).

Mobile Phase Nonpolar Liquid Polar Liquid ( Water / Methanol )

 

Principle :

  • Polar molecue dont react with non polar molecule.
  • Non Polar molecule react with non polar molecule.

Hplc

Many peak available is the chromatogram. There are two types, Qualitative analysis and Quantitative analysis.

Qualitative Analysis

Qualitative analysis is the discovery of how many molecules a sample has on hand.

Quantitative Analysis

Quantitative analysis is the act of knowing how much there is.

Advantages :

  • Efficient in analysis biological samples such proteins , lipids etc.
  • Fast, sensitive, automated and precise.
  • Both Qualitative and Quantitative analysis can be done.

Disadvantages :

  • Expensive
  • Detector is complex.
  • Difficult for novices ( need skill to handle ).

Video :

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