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    High Performance Liquid Chromatography (HPLC)

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    Jitender Yadav
    The document summarizes an experiment using high performance liquid chromatography to separate and analyze sugars. The student aims to learn HPLC operation, optimize sugar separation using isocratic elution, obtain data on retention times, peak widths and areas, calculate resolution and plate number efficiency, and create a calibration curve to quantify unknown concentrations. Key results are resolutions between glucose, xylose and arabinose peaks, and plate numbers for each sugar. The student concludes they gained familiarity with HPLC techniques and parameters.

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    High Performance Liquid Chromatography (HPLC)

    Uploaded by

    Jitender Yadav
    12 views3 pages
    The document summarizes an experiment using high performance liquid chromatography to separate and analyze sugars. The student aims to learn HPLC operation, optimize sugar separation using isocratic elution, obtain data on retention times, peak widths and areas, calculate resolution and plate number efficiency, and create a calibration curve to quantify unknown concentrations. Key results are resolutions between glucose, xylose and arabinose peaks, and plate numbers for each sugar. The student concludes they gained familiarity with HPLC techniques and parameters.

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    HPLC Jitender 10001033

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    The document summarizes an experiment using high performance liquid chromatography to separate and analyze sugars. The student aims to learn HPLC operation, optimize sugar separation using isocratic elution, obtain data on retention times, peak widths and areas, calculate resolution and plate number efficiency, and create a calibration curve to quantify unknown concentrations. Key results are resolutions between glucose, xylose and arabinose peaks, and plate numbers for each sugar. The student concludes they gained familiarity with HPLC techniques and parameters.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd
    Flag for inappropriate content
    Download as pdf or txt
    12 views3 pages

    High Performance Liquid Chromatography (HPLC)

    Uploaded by

    Jitender Yadav
    The document summarizes an experiment using high performance liquid chromatography to separate and analyze sugars. The student aims to learn HPLC operation, optimize sugar separation using isocratic elution, obtain data on retention times, peak widths and areas, calculate resolution and plate number efficiency, and create a calibration curve to quantify unknown concentrations. Key results are resolutions between glucose, xylose and arabinose peaks, and plate numbers for each sugar. The student concludes they gained familiarity with HPLC techniques and parameters.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd
    Flag for inappropriate content
    Download as pdf or txt
    of 3

    Jitender Kumar 10001033

    Experiment No. 6

    High Performance liquid chromatography (HPLC)


    Aim:
    1. Learn the basics of liquid chromatography and to learn how to operate the WATERS 600E HPLC. 2. Optimize the separation of two compounds (sugars) using isocratic elution methods. 3. Obtain following data from EMPOWER (HPLC software): retention time (Tr), peak Widths (w), and peak areas for each peak. 4. Calculate following parameters on the basis of data obtained from HPLC: Resolution between peaks (Rs), and Plate number or efficiency (N) for each peak. 5. Plot the calibration curve for mixture of both compounds and quantify the Concentration of unknown compound

    Observations:

    Jitender Kumar 10001033

    Experiment No. 6

    Jitender Kumar 10001033

    Experiment No. 6

    Results and discussion:


    Resolution between the peaks: Between Glucose and Xylose: 0.013288 Between Xylose and arabinose: 0.015449 Between Arabinose and Glucose: 0.029451 Using the formula:

    Where tr2 and tr1 are the retention times for second component and first component and w 1 and w2 are the width of the peaks. Plate number of efficiency: Glucose: 0.872429 Xylose: 0.4742 Arabinose: 0.364115 The absorption Chromatograph is shown below

    Conclusion:
    Have got familiar with HPLC technique and calculated the peak area and retention time which are basic parameters in HPLC experiment. We got some additional peaks in chromatograph, it may be due to impurities present in solution.

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