Estimation of carbohydrate (By anthrone reagents)

The detection of carbohydrate material by means of anthrone (9-Keto anthra- cene) was introduced by Dreywood in 1946
*Principle:
Carbohydrates are dehydrated with concentrated H2SO4 to form “Furfural”, which condenses with anthrone to form a green color complex which can be measured by using colorimetrically at 620nm (or) by using a red filter. Anthrone reacts with dextrins, monosaccharides, disaccharides, polysaccharides, starch, gums, and glycosides. But they yields of color where is to form carbohydrate to carbohydrate.
Anthrone is a tricyclic sweet-smelling ketone. It is utilized for a well-known cellulose measure and in the colorimetric determination of carbohydrates.
Anthrone is used for a common cellulose assay and in the colorometric determination of carbohydrates. Derivatives of anthrone are used in pharmacy as laxative. They stimulate the motion of the colon and reduce water reabsorption.
Anthrone test is also another general test for all carbohydrates. In this test also, carbohydrate gets dehydrated when react with conc. H2SO4 to form furfural. This furfural reacts with anthrone to give bluish green colored complex.

*Protocol :-
Weigh 100mg of the sample into a boiling tube.
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Hydrolyse by keeping it in boiling water bath for 3 hours with 5mL of 2.5 N-HCl and cool to room temperature.
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Neutralise it with solid sodium carbonate until the effervescence ceases.
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Make up the volume to 100mL and centrifuge.
                 ⬇️
Collect the supernatant and take 0.5 and 1mL aliquots for analysis.
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Prepare the standards by taking 0, 0.2, 0.4, 0.6, 0.8 and 1mL of the working standard. ‘0’ serves as blank.
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Make up the volume to 1mL in all the tubes including the sample tubes by adding distilled water.
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Then add 4mL of anthrone reagent.
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Heat for eight minutes in a boiling water bath.
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Cool rapidly and read the green to dark green color at 630nm.

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