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    Chromatography HPLC

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    Bodea Doru
    This document discusses liquid chromatography and high performance liquid chromatography (HPLC). It provides a brief history of chromatography, describing how it was discovered in 1903 and used to separate plant pigments. It then explains that HPLC provides high speed, efficiency, and sensitivity compared to liquid chromatography. The key components of an HPLC system are described as the pump, injector, column, detector, and computer. Different types of separation columns and general applications of HPLC are also summarized.

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    Chromatography HPLC

    Uploaded by

    Bodea Doru
    140 views13 pages
    This document discusses liquid chromatography and high performance liquid chromatography (HPLC). It provides a brief history of chromatography, describing how it was discovered in 1903 and used to separate plant pigments. It then explains that HPLC provides high speed, efficiency, and sensitivity compared to liquid chromatography. The key components of an HPLC system are described as the pump, injector, column, detector, and computer. Different types of separation columns and general applications of HPLC are also summarized.

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    Liquid chromatography and High Performance Liquid Chromatography

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    This document discusses liquid chromatography and high performance liquid chromatography (HPLC). It provides a brief history of chromatography, describing how it was discovered in 1903 and used to separate plant pigments. It then explains that HPLC provides high speed, efficiency, and sensitivity compared to liquid chromatography. The key components of an HPLC system are described as the pump, injector, column, detector, and computer. Different types of separation columns and general applications of HPLC are also summarized.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    140 views13 pages

    Chromatography HPLC

    Uploaded by

    Bodea Doru
    This document discusses liquid chromatography and high performance liquid chromatography (HPLC). It provides a brief history of chromatography, describing how it was discovered in 1903 and used to separate plant pigments. It then explains that HPLC provides high speed, efficiency, and sensitivity compared to liquid chromatography. The key components of an HPLC system are described as the pump, injector, column, detector, and computer. Different types of separation columns and general applications of HPLC are also summarized.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 13

    Liquid chromatography and High

    Perfomance Liquid Chromatography


    (HPLC)

    By
    LEBECHI A.K.
    Chromatography
     Chromatography was discovered in 1903 and was used
    to separated plant pigments.

     It is a popular method used to separate mixtures to their


    individual components.

    Separate
    Mikhail Semenovich Tswett

    Mixture
    Components
    TYPES OF CHROMATOGRAPHIC TECHNIQUES
    Thin layer chromatography
    COLUMN CHROMATOGRAPHY
    HPLC
     The history of HPLC began in the 60’s and was known as high
    pressure liquid chromatography.

     The instrumentation and columns improved over the time and it is


    now known as high performance liquid chromatography.

     HPLC separation technique provides high speed, efficiency and high


    sensitivity as compared to liquid chromatography.

     A very small volume of sample is injected into a tube packed with a


    stationary phase (3-5 microns particles).

     The sample is pushed down the column with a mobile phase by


    applying high pressure and the components are detected and
    quantified as they exit the column.
    Components of HPLC
    1. Pump: pumps the mobile phase at a specific flow rate in mL/min. The
    pump pressure is normally between 400-600 bar.

    1. Injector: Introduces the sample into the column (about 5-20 μL).

    1. Column: provides separation through high pressure created by the small


    particles.

    2. Detector: it quantifies and identify the sample components and provides


    information to the computer.

    3. Computer: takes the signals from the detector and displays the retention
    times and quantity of the components.
    HPLC system
    Separation columns
     Normal phase: Column packing is polar (e.g silica) and the mobile phase is non
    polar. It is used for water-sensitive compounds, geometric isomers, cis-trans
    isomers and chiral compounds.

     Reverse phase: the column packing is non-polar (e.g C18) , mobile phase is
    water+ miscible solvent (e.g methanol). It can be used for polar, non polar,
    ionizable and ionic samples.

     Ion exchange: Column packing contains ionic groups and the mobile phase is
    buffer. It is used to separate anions and cations.

     Size exclusion: molecules diffuse into pores of a porous medium and are
    separated according to their relative size to the pore size. Large molecules elute
    first and smaller molecules elute later.
    General applications
     HPLC is used in diagnosis of diseases and disorders.
    It is used to estimate metabolites in urine samples and plasma.

     For quality assurance purposes in both food and pharmaceutical


    industries.

     In scientific research for analysis.


    Conclusions
    Chromatography is an effective method of purification.

    The use of HPLC as a support method of analysis could


    explain the fluorescence behavior of the compounds.

     HPLC is not for decoration, lets be more analytical!

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