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  • Reverse Phase Chromatography

    Încărcat de

    Raphael Caballero
    36 vizualizări7 pagini
    Reverse phase chromatography involves separating molecules based on their hydrophobicity using a stationary phase that is hydrophobic or non-polar and a mobile phase that is hydrophilic or polar. Molecules are separated in steps - first the sample binds to the stationary phase, then hydrophilic molecules are eluted followed by hydrophobic molecules as the polarity of the mobile phase is increased. Reverse phase chromatography has advantages like good selectivity and reproducibility but disadvantages like higher costs and time compared to normal phase chromatography which uses a polar stationary phase and nonpolar mobile phase.

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    QC2 Report

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    Reverse phase chromatography involves separating molecules based on their hydrophobicity using a stationary phase that is hydrophobic or non-polar and a mobile phase that is hydrophilic or polar. Molecules are separated in steps - first the sample binds to the stationary phase, then hydrophilic molecules are eluted followed by hydrophobic molecules as the polarity of the mobile phase is increased. Reverse phase chromatography has advantages like good selectivity and reproducibility but disadvantages like higher costs and time compared to normal phase chromatography which uses a polar stationary phase and nonpolar mobile phase.

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    36 vizualizări7 pagini

    Reverse Phase Chromatography

    Încărcat de

    Raphael Caballero
    Reverse phase chromatography involves separating molecules based on their hydrophobicity using a stationary phase that is hydrophobic or non-polar and a mobile phase that is hydrophilic or polar. Molecules are separated in steps - first the sample binds to the stationary phase, then hydrophilic molecules are eluted followed by hydrophobic molecules as the polarity of the mobile phase is increased. Reverse phase chromatography has advantages like good selectivity and reproducibility but disadvantages like higher costs and time compared to normal phase chromatography which uses a polar stationary phase and nonpolar mobile phase.

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    din 7

    REVERSE PHASE

    CHROMATOGRAPHY
    BY:
    CABALLERO, RAPHAEL
    TURNO, SHIELA MAE
    2

    • involves the separation of molecules


    “Hydrophobic on the basis of hydrophobicity.
    Chromatograph
    y”
    3

    Stationary
    Hydrophobic
    Phase or non-polar

    Mobile
    Phase Hydrophilic or
    Polar
    4
    5

    Step 1:
    Starting
    Condition

    Step 5: Step 2:
    Conditioning Binding of
    Phase the Sample

    Step 4: Step 3:
    Elution of Elution of
    Hydrophobic Hydrophilic
    molecules molecules
    6

    ADVANTAGES DISADVANTAGES

    REVERSE PHASE NORMAL PHASE


    •Costly
    •Time consuming
    It uses nonpolar stationary It uses polar stationary • Good selectivity •Requires large amount of
    solvent
    phase and a polar mobile phase and nonpolar mobile • Easily reproducible
    phase phase
    Polar compounds elute Nonpolar compounds elute
    faster than nonpolar faster than polar compounds
    compounds
    When we increase the When we increase the
    polarity of mobile phase polarity of mobile phase
    elution time will decrease elution time will increase
    Stabilize quickly Stabilize slowly and prone to
    fluctuations in retention time
    7

    Good choice for peptides and proteins


    separations when short chain alkyl
    stationary phases are use

    Analysis of Pharmaceuticals,
    Biomolecules & Food products

    First choice for neutral and non-polar


    compounds w/ a MW <2000 Dalton.

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