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    Instrumentation of HPLC

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    umar farooq
    HPLC instrumentation includes a pump, injector, column, detector and data collection system. The pump forces a mobile phase through the column where separation occurs. The injector introduces the sample into the mobile phase stream. Common detectors measure properties like UV absorption, fluorescence or mass. A data collection system integrates the detector response to produce chromatographs for analysis.

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    Instrumentation of HPLC

    Uploaded by

    umar farooq
    3K views2 pages
    HPLC instrumentation includes a pump, injector, column, detector and data collection system. The pump forces a mobile phase through the column where separation occurs. The injector introduces the sample into the mobile phase stream. Common detectors measure properties like UV absorption, fluorescence or mass. A data collection system integrates the detector response to produce chromatographs for analysis.

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    instrumention and the component of hplc

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    instrumentation of hplc

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    HPLC instrumentation includes a pump, injector, column, detector and data collection system. The pump forces a mobile phase through the column where separation occurs. The injector introduces the sample into the mobile phase stream. Common detectors measure properties like UV absorption, fluorescence or mass. A data collection system integrates the detector response to produce chromatographs for analysis.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    3K views2 pages

    Instrumentation of HPLC

    Uploaded by

    umar farooq
    HPLC instrumentation includes a pump, injector, column, detector and data collection system. The pump forces a mobile phase through the column where separation occurs. The injector introduces the sample into the mobile phase stream. Common detectors measure properties like UV absorption, fluorescence or mass. A data collection system integrates the detector response to produce chromatographs for analysis.

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    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
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    Instrumentation of HPLC

    As shown in the schematic diagram in Figure above, HPLC instrumentation includes a pump,
    injector, column, detector and integrator or acquisition and display system. The heart of the
    system is the column where separation occurs.

    1. Solvent Resorvoir
    Mobile phase contents are contained in a glass resorvoir. The mobile phase, or solvent, in
    HPLC is usually a mixture of polar and non-polar liquid components whose respective
    concentrations are varied depending on the composition of the sample.

    2. Pump
    A pump aspirates the mobile phase from the solvent resorvoir and forces it through the
    system’s column and detecter. Depending on a number of factors including column
    dimensions, particle size of the stationary phase, the flow rate and composition of the mobile
    phase, operating pressures of up to 42000 kPa (about 6000 psi) can be generated.
    3. Sample Injector
    The injector can be a single injection or an automated injection system. An injector for an
    HPLC system should provide injection of the liquid sample within the range of 0.1-100 mL
    of volume with high reproducibility and under high pressure (up to 4000 psi).

    4. Columns
    Columns are usually made of polished stainless steel, are between 50 and 300 mm long and
    have an internal diameter of between 2 and 5 mm. They are commonly filled with a
    stationary phase with a particle size of 3–10 µm.

    Columns with internal diameters of less than 2 mm are often referred to as microbore
    columns. Ideally the temperature of the mobile phase and the column should be kept constant
    during an analysis.

    5. Detector
    The HPLC detector, located at the end of the column detect the analytes as they elute from
    the chromatographic column. Commonly used detectors are UV-spectroscopy, fluorescence,
    mass-spectrometric and electrochemical detectors.

    6. Data Collection Devices


    Signals from the detector may be collected on chart recorders or electronic integrators that
    vary in complexity and in their ability to process, store and reprocess chromatographic data.
    The computer integrates the response of the detector to each component and places it into a
    chromatograph that is easy to read and interpret.

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