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    Determination of Total Carbohydrates by Anthrone Method

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    Guru Raj PN
    This document describes the anthrone method for determining total carbohydrates in a sample. It involves hydrolyzing carbohydrates into simple sugars like glucose using acid hydrolysis. Glucose then reacts with anthrone reagent to form a green colored compound that can be measured spectrophotometrically. By comparing to a standard glucose curve, the amount of original carbohydrates in the sample can be calculated. The procedure involves hydrolyzing the sample, neutralizing it, taking aliquots, preparing standards, adding anthrone reagent, heating, measuring absorbance, and using the standard curve to determine total carbohydrates in the original sample.

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    Determination of Total Carbohydrates by Anthrone Method

    Uploaded by

    Guru Raj PN
    153 views2 pages
    This document describes the anthrone method for determining total carbohydrates in a sample. It involves hydrolyzing carbohydrates into simple sugars like glucose using acid hydrolysis. Glucose then reacts with anthrone reagent to form a green colored compound that can be measured spectrophotometrically. By comparing to a standard glucose curve, the amount of original carbohydrates in the sample can be calculated. The procedure involves hydrolyzing the sample, neutralizing it, taking aliquots, preparing standards, adding anthrone reagent, heating, measuring absorbance, and using the standard curve to determine total carbohydrates in the original sample.

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    Determination of Total Carbohydrates

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    Determination of total carbohydrates by anthrone method

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    This document describes the anthrone method for determining total carbohydrates in a sample. It involves hydrolyzing carbohydrates into simple sugars like glucose using acid hydrolysis. Glucose then reacts with anthrone reagent to form a green colored compound that can be measured spectrophotometrically. By comparing to a standard glucose curve, the amount of original carbohydrates in the sample can be calculated. The procedure involves hydrolyzing the sample, neutralizing it, taking aliquots, preparing standards, adding anthrone reagent, heating, measuring absorbance, and using the standard curve to determine total carbohydrates in the original sample.

    Copyright:

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

    Determination of Total Carbohydrates by Anthrone Method

    Uploaded by

    Guru Raj PN
    This document describes the anthrone method for determining total carbohydrates in a sample. It involves hydrolyzing carbohydrates into simple sugars like glucose using acid hydrolysis. Glucose then reacts with anthrone reagent to form a green colored compound that can be measured spectrophotometrically. By comparing to a standard glucose curve, the amount of original carbohydrates in the sample can be calculated. The procedure involves hydrolyzing the sample, neutralizing it, taking aliquots, preparing standards, adding anthrone reagent, heating, measuring absorbance, and using the standard curve to determine total carbohydrates in the original sample.

    Copyright:

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

    Determination of total carbohydrates by anthrone method

    Carbohydrates are the important components of storage and structural materials in the plants.
    They exist as free sugars and polysaccharides. The basic units of carbohydrates are the
    monosaccharides which cannot be split by hydrolysis into more simpler sugars. The
    carbohydrate content can be measured by hydrolyzing the polysaccharides into simples sugars by
    acid hydrolysis and estimating the resultant monosaccharides.

    Principle
    Carbohydrates are first hydrolysed into simple sugars using dilute hydrochloric acid. In hot
    acidic medium glucose is dehydrated to hydroxymethyl furfural. This compound forms with
    anthrone a gree colored product with an absorption maximum at 630nm.

    Materials
    ⇒ 2.5 N-HCl

     Anthrone Reagent: Dissolve 200mg anthrone in 100mL of ice cold 95% H2SO4. Prepare
    fresh before use.
     Standard Glucose: Stock – Dissolve 100mg in 100mL water. Working standard – 10mL
    of stock diluted to 100mL with distilled water. Store refrigerated after adding a few drops of
    toluene.

    Procedure

    1. Weigh 100mg of the sample into a boiling tube.


    2. Hydrolyse by keeping it in boiling water bath for 3 hours with 5mL of 2.5 N-HCl and
    cool to room temperature.
    3. Neutralise it with solid sodium carbonate until the effervescence ceases.
    4. Make up the volume to 100mL and centrifuge.
    5. Collect the supernatant and take 0.5 and 1mL aliquots for analysis.
    6. Prepare the standards by taking 0, 0.2, 0.4, 0.6, 0.8 and 1mL of the working standard. ‘0’
    serves as blank.
    7. Make up the volume to 1mL in all the tubes including the sample tubes by adding
    distilled water.
    8. Then add 4mL of anthrone reagent.
    9. Heat for eight minutes in a boiling water bath.
    10. Cool rapidly and read the green to dark green color at 630nm.
    11. Draw a standard graph by plotting concentration of the standard on the X-axis versus
    absorbance on the Y-axis.
    12. From the graph calculate the amount of carbohydrate present in the sample tube.

    Calculation
    Amount of carbohydrate present in 100mg of the sample =(mg of glucose ÷ Volume of test
    sample) X 100
    Note
    Cool the contents of all the tubes on ice before adding ice-cold anthrone reagent.

    Reading

    1. Hedge, J E and Hofreiter, B T (1962) In: Carbohydrate Chemistry 17 (Eds Whistler R L


    and Be Miller, J N) Academic Press New York.

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