Experiment 6: Carbonyl Compounds and Carbohydrates Results and Discussions The carbonyl group, C=O is common to many families

of compounds. The carbonyl carbon atom bears a partial positive charge and the oxygen bears a partial negative charge. On the first part of the experiment, benzaldehyde (an aldehyde), acetone (a ketone), glucose (hemiacetal) and starch (polymer of glucose) were used. Aldehydes have at least one hydrogen bonded to the C=O while ketones have two carbons bonded to the C=O. Glucose exists as hemiacetals due to an intermolecular reaction involving the C=O group and one of the OH groups in the chain. Since all of these groups contain the carbonyl group, they are all polar and are therefore soluble in water. Benzaldehyde in fact however is slightly soluble with the polar solvent. This slight solubility of benzaldehyde with water is accounted from the stability acquired by the said compound due to electron delocalization or electronegativity sharing within its cyclic structure. In the hydrolysis of Di- and Polysaccharide, water molecule is introduced to the glycosidic bond, thus breaking the polysaccharide into its monosaccharide units. The three samples were sucrose, cellulose and starch. Upon addition of water, they were broken into their monosaccharide units. Sucrose is common table sugar. It is composed of a - D- Glucose and - D- fructose.

Cellulose is the chief structural materal of plants. It is a linear polymer of - D- glucose.

Starch is the storage form of glucose in plants. It is a polymer of glucose but the type of glycosidic linkage differs from that of cellulose. Starch is made up of two components: amylase which has a linear structure and amylopectin, having a highly-branched structure. Amylose and amylopectin contain a-D-glucose units.

Aldehydes are easily oxidized to yield carboxylic acids, RCHO RCO2H but ketones are unreactive towards oxidation. This reactivity difference is a consequence of structure; aldehydes have a CHO proton that can be removed during oxidation, but ketones do not.

One of the simplest methods for oxidizing an aldehyde is to use silver ion, Ag+, in dilute aqueous ammonia, a mixture called the Tollens reagent. As the oxidation proceeds, a shiny mirror of silver metal is deposited on the walls of the reaction flask, forming the basis of the differentiation between aldehydes and ketones. In the experiment, a silver mirror formed on the acetaldehyde but not on the acetone.

The next part involves the iodoform test. The basis of this test is the ability of certain compounds to form a preciptate of iodoform when treated with basic solution of iodine. This is usually performed to distinguish methyl ketones from other ketones. The methyl ketone is treated with iodine in a basic solution. Introduction of the first iodine atom increases the acidity of the remaining methyl protons. A subsequent proton transfer gives iodoform, a yellow crystalline solid. In the experiment, cyclohexanone and ketone, which are both ketones were subjected to this test. Comparison of their structures show however that acetone is a methyl ketone.

Thus, it is expected that a yellowish precipitate will form when acetone is reacted with the I2/KI solution. CH3COCH3 + I2 --> CH3COCH2I + H+ +IAs can be observed from the results, there was a precipitate in the acetone but none in the cyclohexanone.

After this, acetone and acetaldehyde are reacted with 2,4 DNP. -test for aldehydes and ketones. The positive result for this test is the formation of a yellow orange precipitate. From the data, both the acetaldehyde and acetone formed a precipitate. The last part of the experiment was the reactions of carbohydrates. The carbohydrates have structural as well as metabolic functions in both animal and plant tissues. In animal tissues they are in the form of glucose and glycogen while in plant tissues they occur as starch, sugars and cellulose. Chemically they are aldehyde or ketone derivatives of the polyhydric alcohols and compounds which can be converted into such aldehydes or ketones by hydrolysis. A number of reactions are used in the characterization of carbohydrates. They are grouped according to the chemical reaction involved, namely, the fragmentation of carbohydrates by strong mineral acids, the reduction of certain metabolic ions by sugars, the oxidation of carbohydrates by concentrated mineral acids and the hydrolysis of carbohydrates by weak acids and enzymes. The Molisch Alpha Naphthol reaction is a general test for carbohydrates in free or combined from. In the presence of concentrated H2SO4, glycosidic bonds are hydrolyzed giving monosaccharides which are then dehydrated to furfural, hydroxymethyl- furfural and other decomposition products. These react the alpha naphthol forming purple-colored condensation products at the zone junction of the two liquids. Concentrated solutions of organic may give a red instead of a violet color due to the charring action of H2SO4. In such a case, the reaction should be repeated on amore dilute solution of te specimen. The test is not a specific test for carbohydrates since it is also given by furfural and other furfural-yielding substances. However, a negative result is good evidence of the absence of carbohydrate (The Biochemistry Faculty, 1980). The reaction with a pentose is represented in these equations:

Basing on our data, starch, glucose and sucrose formed a purple layer between the junctions of the two liquids. There was none in the cyclohexanone and this means that there is no carbohydrate. The reducing properties of sugars are dependent on the presence of an actual or potential, aldehyde or ketone group in the molecule. When solutions of such sugars as the monosaccharides and the disaccharides maltose and lactose are heated in the presence of certain metallic ions are reduced to a lower valence state. The reagents used are solutions of metallic salts of copper, mercury, bismuth, iron, gold, or silver, usually in an alkaline medium, mixed with organic compounds containing alcoholic groups. The most reliable and the most widely used salt is copper. Benedicts reagent contains CuSO4, Na2CO3 and Ca3(C6H5O7)2 4H2O (calcium nitrate). The reactions are shown in the following equations:

The cupric ion is reduced and the resulting cuprous oxide precipitates out of the alkaline solution as a brick red solid. From the data, it can be inferred that the reagent Cu2+ reacted with the carbonyl group of carbohydrates to form a red precipitate of cuprous oxide. The ketoses (fructose) also reacted with benedicts reagent. Reducing sugars which have either a free aldehyde or a ketone group will react with phenylhydrazine to form osazones. With an excess of phenylhydrazine, the carbon adjacent to the aldehyde or ketone group is oxidized to an aldehyde or ketone which also condenses with phenylhydrazine to form a yellow crystalline compound or osazone. These yellow compounds are valuable for the identification of sugars for the following reasons: a. the time formation of each osazone is quite definite, b. some osazones are relatively insoluble in hot water while others are soluble, c. each osazone, if carefully purified by recrystallization and dried, has a definite melting point and , d. osazones crystallize in characteristic forms which could be appreciated by microscopic examination and reference to standard osazones. Although each individual sugar should give rise to an osazone typical for that sugar, glucose, glucosamine, fructose and mannose yield the same osazone because of similarities in their molecular structure. The reaction with glucose and maltose resulting in the formation of osazones are represented thus:

In the experiment the first to form osazone is the fructose, glucose and lastly the sucrose. The lactose took a very long time to form a precipitate. III. Conclusion The experiment included various tests for the identification and differentiation of aldehydes, ketones and carbohydrates. The Tollens test is the the basis of the differentiation between aldehydes and ketones. Aldehydes forms a shiny mirror of silver metal at the walls of the tube while ketone does not. Iodoform on the other hand, is performed to distinguish methyl ketones from other ketones. Methyl ketones can form a preciptate of iodoform, a yellow crystalline solid when treated with basic solution of iodine. Reaction with 2,4- DNP differentiates aldehydes and ketones from other compounds through the formation of a yellow to orange precipitate. In carbohydrates, the Molisch Alpha Naphthol reaction is a general test for carbohydrates in free or

combined from. The formation of purple-colored condensation at the zone junction of the two liquids, indicate the presence of a carbohydrate. Benedicts reagent tests the reducing properties of sugars are dependent on the presence of an actual or potential, aldehyde or ketone group in the molecule. Also, these reducing sugars which have either a free aldehyde or a ketone group will react with phenylhydrazine to form osazones which is also a factor in differentiating compounds. IV. Answers to questions 1. The carbonyl group, C=O is common to many families of compounds. The carbonyl carbon atom bears a partial positive charge and the oxygen bears a partial negative charge. On the first part of the experiment, benzaldehyde (an aldehyde), acetone (a ketone), glucose (hemiacetal) and starch (polymer of glucose) were used. Aldehydes have at least one hydrogen bonded to the C=O while ketones have two carbons bonded to the C=O. Glucose exists as hemiacetals due to an intermolecular reaction involving the C=O group and one of the OH groups in the chain. Since all of these groups contain the carbonyl group, they are all polar and are therefore soluble in water. 2. Only carbonyl compounds possessing the structure can undergo the haloform reaction. Only these compounds tend to give a positive result to the haloform reaction. 3. Basing on the structure of aldehydes and ketones, aldehydes actually have the advantage of releasing the H atom attached to the carbonyl group. Thus, aldehydes are reducing agents, hence easily oxidized. Ketones in one aspect are also oxidizable only that they require stronger oxidizing agents because they do not have H atom bonded to the carbonyl group. The oxidation in ketones also involve breaking of the C=C bond. Thus, ketones rather are oxidizable compounds. 4. Benedicts test is a chemical test for reducing sugars or those sugars which reduce metallic oxidizing agent. Among the sugars, sucrose yielded the negative result since it is not itself a reducing sugar. That is, the potential carbonyl group of the second monosaccharide unit of this disaccharide is tied up in the glycosidic linkage, making reactions due to the free carbonyl group not possible. The hydrolysate of sucrose however is positive with the Benedicts test because hydrolysis freed its constituent and bound glucose (-D-glucose) and fructose, both of which are reducing sugars. Starch on one aspect is a reducing sugar. As seen from its structure, the potential carbonyl group of the second monosaccharide unit is not attached or involved in the glycosidic linkage. Thus, it exists in equilibrium with its free carbonyl form and can undergo reactions due to such group. Hydrolysis of starch actually results to -D-glucose units, which are reducing sugars and thus gives a positive result for the starch hydrolysate.

V. References Laboratory Manual in Chemistry. Department of Nutrition. College of Public Health. UP Manila. Philippines. pp. 3-14. McMurry, J. Fundamentals of Organic Chemistry. 6th edition. 2008. Thomson Brooks Cole. USA.