Color reactions of carbohydrate:

Experiment Observation Remarks

1. Fehlings Test Formation of yellow or To 1mL of Fehlings solution A, add 1mL of brownish-red precipitate Fehlings solution B and a few drops of the test solution. Boil for a few minutes.

The blue alkaline cupric hydroxide present in solution, when heated in the presence of reducing sugars, gets reduced to yellow or red cuprous oxide and it gets precipitated. Hence, formation of the colored precipitate indicates the presence of reducing sugars in the test solution. 2. Benedicts Test Formation of red, yellow or As in Fehlings test, the reducing To 2mL of Benedicts reagent add five drops green color/precipitate. sugar because of having potentially of the test solution. Boil for five minutes in a free aldehyde or keto group reduce water bath. Cool the solution. cupric hydroxide in alkaline solution to red colored cuprous oxide. Depending on the sugar concentration yellow to green color is developed. Barfoeds Test Formation of brick-red Only monosaccharides answer this 3. To 1mL of the test solution add about 2mL precipitate. test. Since Barfoeds reagent is of Barfoeds reagent. Biol it for one minute weakly acidic, it is reduced only by and allow to stand for a few minutes. monosaccharides.

How is the brick-red coloration formed by Benedict's reagent in the prescence of reducing sugars?
One of the components of Benedict's reagent is copper (II) oxide.when it is placed in solution with a reducing sugar, the copper (II) oxide is reduced to copper (I) oxide ions. This has a brick red color. If there a small or large amount of the reducing sugar present, the color would range from green to brick red.

Fehlings test:
Fehling I consists of 7 g of hydrated copper (II) sulfate dissolved in 100 mol of dist. water. Fehling II is made by dissolving 35 g of potassium sodium tartrate and 10 g of sodium hydroxide in 100 mol of dist. water. Fehling's reagent: Equal volumes of Fehling I and Fehling II are mixed to form a deep blue solution.

Experimental procedure:
5 g of glucose, fructose and sucrose, respectively, are dissolved in 100 mL of dist. water. Three 500 ml conical measures are each filled with the sugar solutions. Each solution is made up to the 500 mL mark with dist. water warmed up to 60 C. Afterwards 8 mL of Fehling reagent are poured

into each of the sugar solutions while stirring.

Results:
In two conical measures a yellow-red precipitate is formed. At first in the blue solution the precipitate may appear greenish. Finally the muddy green suspension yields brick red precipitate. Fructose reacts faster than glucose. In the third glass no change can be observed. conical measure 1 glucose solution brick red precipitate brick red precipitate no change

conical measure fructose solution 2 conical measure 3 sucrose solution

Discussion:
glucose, a reducing sugar, reacts with Fehling's reagent to form an orange to red precipitate. Fehling's reagent is commonly used for reducing sugars but is known to be not specific for aldehydes. For example, fructose gives a positive test with Fehling's solution too, because fructose is converted to glucose and mannose under alkaline conditions. The conversion can be explained by the keto-eno The sucrose does not react with Fehling's reagent. Sucrose is a
disaccharide of glucose and fructose. Most disaccharides are reducing sugars, sucrose is a notable exception, for it is a non-reducing sugar. The anomeric carbon of glucose is involved in the glucose-fructose bond and hence is not free to form the aldehyde.

Table: Phytochemical constituents of exudates of Pycanthus angolensis Test Test for reducing sugars in glycosides (Fehling's solutions test) Frothing test for Saponins Borntrager's reagent test for anthracene derivatives Keller Kiliani test for deoxysugars in cardiac Observation Blue color of Fehling's solutions turned brick red ppt. Only bubbles seen. No frothing Absence of Saponins Same shade of pink as the Absence of Anthracene exudates No brown ring color derivatives Absence of deoxysugars, No cardiac glycosides. Inference Presence of reducing sugar

glycosides Hydrolysable FeCL3 solution test for Tannins Test for flavonoids NaOH Test Blue-black ppt Flavonoids dissolved giving Yellow color. solution turns colorless with HCI test Tests with Dragendorff's, alkaloidal Reagents for Alkaloids HCI No Coloured ppt with any Absence of Alkaloids Flavonoids present. Tannins present

Hager's,Mayer's, Wagner's reagent