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    Estimation of Reducing Sugar by Nelson-Somogyi Method

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    liska ramdanawati
    The document describes the Nelson-Somogyi method for quantitatively determining reducing sugars. It involves using reducing sugars' ability to reduce copper sulfate in an alkaline solution, followed by a colorimetric reaction between the reduced copper and arsenomolybdate to form a blue complex, which is measured spectrophotometrically. Reagents and procedures are provided for preparing copper and Nelson reagents, running standards and samples by boiling with copper reagent then adding Nelson reagent, and measuring absorbance at 520nm.

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    Estimation of Reducing Sugar by Nelson-Somogyi Method

    Uploaded by

    liska ramdanawati
    4K views2 pages
    The document describes the Nelson-Somogyi method for quantitatively determining reducing sugars. It involves using reducing sugars' ability to reduce copper sulfate in an alkaline solution, followed by a colorimetric reaction between the reduced copper and arsenomolybdate to form a blue complex, which is measured spectrophotometrically. Reagents and procedures are provided for preparing copper and Nelson reagents, running standards and samples by boiling with copper reagent then adding Nelson reagent, and measuring absorbance at 520nm.

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    Estimation of Reducing Sugar by Nelson-somogyi Method

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    Estimation of Reducing Sugar by Nelson-somogyi Method

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    The document describes the Nelson-Somogyi method for quantitatively determining reducing sugars. It involves using reducing sugars' ability to reduce copper sulfate in an alkaline solution, followed by a colorimetric reaction between the reduced copper and arsenomolybdate to form a blue complex, which is measured spectrophotometrically. Reagents and procedures are provided for preparing copper and Nelson reagents, running standards and samples by boiling with copper reagent then adding Nelson reagent, and measuring absorbance at 520nm.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    4K views2 pages

    Estimation of Reducing Sugar by Nelson-Somogyi Method

    Uploaded by

    liska ramdanawati
    The document describes the Nelson-Somogyi method for quantitatively determining reducing sugars. It involves using reducing sugars' ability to reduce copper sulfate in an alkaline solution, followed by a colorimetric reaction between the reduced copper and arsenomolybdate to form a blue complex, which is measured spectrophotometrically. Reagents and procedures are provided for preparing copper and Nelson reagents, running standards and samples by boiling with copper reagent then adding Nelson reagent, and measuring absorbance at 520nm.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    of 2

    Estimation of reducing sugar by NelsonSomogyi method

    Micha Kaczmarek

    Keywords: reducing sugars, Somogyi-Nelson assay,


    equipment required:
    spectrophotometer capable of measuring
    absorbance in the 520 nm region.
    cuvettes for spectrophotometer
    water bath 100C
    required reagents:
    carbohydrate standards (glucosamine) at the concentration ranging from 0
    mol/ml to 0,50 mol/ml
    Copper reagent :
    180 g Na2SO4
    12 g potassium sodium tartrate
    24 g Na2CO3 (anhydrous)
    4 g CuSO4 x 5 H2O lub 2,6 g CuSO4
    16 g NaHCO3
    Preparation:
    Dissolve 12 g of potassium sodium tartrate and 24 g Na 2CO3 in 250 ml H2O.
    Followed by continuous stirring add 4 g CuSO4 x 5 H2O i 16 g NaHCO 3.
    Dissolve 180g of Na2SO4 in 500ml of boiling water H2O and boil to remove
    air bubbles.
    The two solutions were mixed and filtered hot to obtain a clear solution.
    Nelson reagent:
    25 g (NH4)2MoO4 x 2H2O or 25,75 g (NH4)6Mo7O24 x 4H2O
    21 cm3 H2SO4
    3g AsHNa2O4 x 7 H2O
    Preparation :
    Dissolve 25 g (NH4)2MoO4 x 2H2O or 25,75 g (NH4)6Mo7O24 x 4H2O in 450
    cm3 H2O and add 21 cm3 H2SO4 (concentrated)
    Dissolve 3g AsHNa2O4 x 7 H2O in 25 cm3 H2O
    Mix both reagents and incubate 30 min. in 55C
    execution time: from 10 min
    Procedure Description:
    The Nelson-Somogyi method is one of the classical and widely used
    methods for the quantitative determination of reducing sugars. This method
    utilizes the reducing properties (because of the presence of a potential aldehyde
    or keto-groups) of certain types of carbohydrates. Determination of reducing

    sugars using Somogyi-Nelson is based on the absorbance at 520 nm of a


    coloured complex between a copper oxidized sugar and arsenomolybdate.

    Procedure:
    1. Prepare carbohydrate standards in distilled water ranging from 00,50 mol/ml.
    2. Add 1 ml of each standard to separate tubes. To the tubes used as
    the blanks, add 1 ml of distilled water.
    3. Prepare the unknown samples in an appropriate dilution.
    4. To each tubes, add 1 ml of the Copper Reagent and mix.
    5. Place the tubes in a boiling water for 10 minutes
    6. Cool the tubes to room temperature and add 1mL of
    arsenomolybdate
    reagent Nelson Reagent to all the tubes.
    7. Measure the absorbance at 520 nm.

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