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    Assignment No. 2
    CHE301 – Analytical Chemistry & Instrumentaions
    Fall 2019

    Total Marks: 10
    Due date: 16-01-2020
    • Make sure that you upload the solution file before due date. No assignment will be accepted through e-mail after the due date.
    Formatting guidelines
    • Use the font style “Times New Roman” and font size “12”.
    • Compose your document in MS-Word, any file created in any other format will not be accepted and assigned zero mark.
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    Solution guidelines
    • To solve this assignment, you should have good command over lectures 64-74.
    • This is not a group assignment, it is an individual assignment so be careful and avoid copying others’ work
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    Please note that your assignment will not be graded if:
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    Question No. 1
    Discuss Instrumentation & working of HPLC? (10)

    ---------------------------------------------------GOOD LUCK----------------------------------------------

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    @Arbaz-Kaleem Solution idea posted

  • Cyberian's Gold

    @zareen said in CHE301 Assignment 2 Solution and Discussion:


    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.

  • Cyberian's Gold

    @zareen said in CHE301 Assignment 2 Solution and Discussion:

    Discuss Instrumentation & working of HPLC?

  • @zareen assignment solution plz

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