Expt.

5: Analysis of Carbohydrates

Janine Manjares Mendato Marcaban* Carl Monzon Department of Psychology, University of Santo Tomas, Manila, Philippines

Abstract The objectives of this experiment is to classify a given sugar as monosaccharide or dissacharide, to to described the structural feature common to reducing sugars , and to be able to deduce the identity of an unknown sugar samples. There are six set-ups; they are the Barfoeds test, Benedicts test , Bials Orcinol test , Seliwanoff test and Iodine test. The first set up is the Barfoeds test because it is a test for determining the monosaccharide for disaccharide, then the Bials Orcinols test is for determining the pentose sugars, Seliwanoff is for classifying ketohexoses then the benedicts test is for classifying the reducing sugars , and Iodine test for helical polysaccharide. In the end the 3 unknowns that given to us are the xylose , glucose and sucrose.

Introduction Carbohydrates occur in every living organism. The sugars and starch infood and the cellulose in wood, paper, and cotton are nearly pure carbohydrate. The word carbohydrate derives historically from the that glucose, the first carbohydrate to be obtained pure and because of that it originally thought it is to be a hydrate of carbon but the term is abandon today. Carbohydrates are referring loosely in sugars. Carbohydrates are generally classed into two groups: simple and complex. Simple sugars or monosaccharide, are carbohydrates like glucose and fructose that cant be converted into smaller sugars by hydrolysis. Complex carbohydrates are

composed of two or more sugars linked together. Sucrose (table sugar), for example is a disaccharide made up of one glucose molecule linked to one fructose molecule. The objectives in the end is to classify a given sugar either as monosaccharide or disaccharide, to describe the structural feature common to reducing sugars, and to be able to deduce the identity of an unknown sugar sample.

Results and Discussions The experiment is compose of 5 set-ups which are the Barfoeds test , Bials Orcinols test , Benedicts test , Seliwanoff test n and the Iodine test. In the Benedicts Test a reducing sugar (a sugar with a free or potentially free, i.e., a cyclic hemiacetal, aldehyde group) reacts with the blue-colored Cu2+ ion in the presence of base The copper (II) ion is reduced to a red-orange Cu2O precipitate whereas the aldehyde group is oxidized to the carboxylic acid functional group. In addition to all aldose monosaccharides giving a positive Benedicts Test, ketose monosaccharides, though lacking an aldehyde group,react due to the presence of an hydroxyl group next to the ketone group. Thus -hydroxy ketones give positive tests. If there is no potential free aldehyde group, i.e., the aldehyde group is tied up in a glycosidic bond (an acetal bond), the sugar is referred to as a non-reducing sugar. The Barfoeds Test is a variation of the redox reaction mentioned previously. Copper (II) acetate in acetic acid is not as reactive as the Cu2+ Benedicts reagent. Thus, one can distinguish monosaccharides from disaccharides based on how fast the red-orange precipitate forms. Typically, monosaccharides react within 2-3 minutes, whereas disaccharides take longer. To distinguish pentoses from hexoses on can use the Bials Test. Pentoses react with orcinol in the presence of FeCl3 and concentrated HCl to give a characteristic bluegreen color. Nonreacting sugars may produce a brown precipitate but the solution usually remains the yellow color of the FeCl3. The Seliwanoff Test is used to distinguish ketoses from aldoses using the aromatic alcohol in the presence of concentrated hydrochloric acid. This is useful for both monosaccharide ketoses as well as disaccharide ketoses. A positive test is noted by a red colored solution; a yellowish straw or apricot color is not indicative of a positive test. In this experiment we are given 3 unknowns and we label it as A(25),B(26), C(27). In the first step of the experiment we do the Barfoeds test to be able to classify the monosaccharides from the disaccharide and the results were the test tube A is

negative and the test tube B and C is positive. Then we do the the Bials Orcinols test to determine which is the pentose sugar and the results were test tube B is only positive in this test and we simply identify it as a xylose sugar and the others were negative on this test. The third step we do the seliwanoffs test to determine the ketohexoses and the results were sample A is negative and sample C is positive. And the last step to determine the two products was the Benedicts test to identify which are the reducing sugars and in the results were sample A is positive and sample C is negative. Table of results of the test Sample Barfoeds test (-) (+) (+) Bials Orcinols test (-) (+) (-) Seliwanoffs test (-) (+) Benedict test (+) (-) Mucic Acid N/A N/A N/A Iodine test

1.Sample A 2.Sample B 3.Sample C Legend

N/A N/A N/A

(+)-positive in the test (-)- negative in the test In the test given test we identife the 3 samples that given to us. Sample A is glucose, Sample B is xylose and Sample C is sucrose. Chemical formula of the unknown

Figure 1. Glucose

Figure 2. Xylose

Figure 3. Sucrose

In this experiment common error is that in other test is that you will have the false result like in the Barfoeds test because if you didnt get it early in the boiling you may get even the maltose positive because the maltose will give positive result if it is boiled longer. Then in the seliwanoffs test you should wait until the color becomes cherry red to have good results.

Refferences
Brown T., Lemay H., Chemistry the Central Science, (2003) pp. 181

Mcmurry J., Fundamentals of Organic Chemistry, (2003) pp 442

Wade L., Organic Chemistry, (1991) pp. 1044 Retrieved on Jan 8/2012 at http://facstaff.bloomu.edu/pugh/Experiment%209.pdf