Title: Macromolecules Objective(s):1. To define monosaccharide, disaccharide, and polysaccharide and give example of each. 2.

To name the monosaccharide components of sucrose and starch. 3. To describe the test that indicates the presence of most small sugars. 4. To describe the test that indicates the presence of starch. 5. To define hydrolysis and give an example of the hydrolysis of carbohydrate. Introduction There are four classes of macromolecules that constitute all living matter: carbohydrates, lipids, proteins, and nucleic acids. While they have different structures and functions, they are all composed of long complex chains of molecules (polymers) made up of simpler, smaller subunits (monomers). They are joined together in a process known as dehydration synthesis, in which a covalent bond is formed between two monomers by releasing a water molecule.Carbohydrates are made up of sugars and their polymers. Simple sugars (monosaccharides) are hydrocarbon chains of varying length that possess a hydroxyl (OH) group on each carbon. Lipids fall into three main categories: fats, steroids, and phospholipids. A fat is composed of a glycerol molecule (a short hydrocarbon) bonded to three fatty acids through dehydration synthesis. These fatty acids are long, nonpolar, hydrocarbon chains, responsible for fats' insolubility in water. Proteins have many levels of structure. Their primary level of structure is the sequence of amino acids linked together in a peptide chain. There are only 20 amino acids, each with a hydrogen, an amino group (NH2 -), a carboxyl group (COO -), and an R group. Nucleic acids include DNA (deoxyribonucleic acid) and RNA (ribonucleic acid). Each is composed of a nitrogenous base, a fvie-carbon sugar (a pentose), and a phosphate group. There are five nitrogenous bases: adnine, guanine, thymine, uracil, and cytosine. It is important to remember that cytosine will only bond with guanine and that adnine will only bond with thymine.

Part 1: Carbohydrates Investigation 1: Monosaccharides and Disaccharides Materials Benedicts solution, 1 % solutions of glucose, fructose, sucrose, lactose and starch, test tube, beaker, measuring cylinder (5ml), Sample A= Potatoes, Sample B= Soft drink, Sample C= Onions, Sample D= Instant noodles, Sample E= Soya beans Method :

1. 1ml of 1% of glucose solution and 5ml of Benedicts solution was added in a test tube. 2. A control was prepared by adding 1ml of distilled water and 5ml of Benedicts solution in a test tube. 3. The test tubes were placed in the water bath at 67C for 2-3 minutes. 4. The color and whether there is precipitate formed was observed. 5. Step 1-4 was repeated with 1% solutions of fructose, lactose, sucrose, starch, and food sample A to E. 6. All of the results were recorded in a table. Investigation 2: Starch Materials Starch solution, Iodine reagent, test tubes, Sample A= Potatoes, Sample B= Soft drink, Sample C= Onions, Sample D= Instant noodles, Sample E= Soya beans Methods :

1. 2ml of starch was added in a test tube. 2. 2 drops of iodine solution was added in the test tube. 3. The change of the color in the solution was noted. 4. 2ml of distilled water was added in another test tube and 2 drops of iodine solution was added in the test tube. 5. The color obtained was noted. 6. Step 1 to 5 was repeated with food sample A to E. 7. Results were recorded.

Investigation 3: Hydrolysis of Carbohydrates Material Starch solution, sucrose solution, 2N HCl solution, Benedicts reagent, Iodine solution, test Tubes, boiling tubes, pipettes, water bath Methods :

1. 5ml of sucrose solution and 1ml of HCl was added in a boiling tube. The tube was labeled as tube A. 2. 6ml of starch solution and 3 ml of HCl solution was added in a second boiling tube and labeled as tube B. 3. 8 test tubes were lined up in the test tube rack and labeled as tube 1-8 respectively. 4. 1ml of solution from tube A was added in tube 1. 5. 1ml of solution from tube B was added in tube 3 and 4. 6. Both tube A and B was kept in the water bath. 7. After 2-3 minutes, tube A was taken out from the water bath and 1ml of the solution was added in tube 2. Tube A was kept in the water bath again. 8. After 6-7 minutes, tube B was taken out from the water bath and 1ml of solution was added in tube 5 and 6. Tube B was kept in the water bath again. 9. After 10 minutes, tube B was taken out again and 1ml of solution was added in tube 7 and 8. 10. 5ml of Benedicts solution was added in tube 1, 2, 3, 5, and 7. These tubes were kept in the water bath for 2-3 minutes. Color changes were observed. 11. 3 drops of iodine solution was added in tube 4, 6 and 8. The color changes were observed. 12. All results were recorded in a table.

Part 2: Proteins Materials 1% egg albumin, concentrated KOH, 0.5% CuSO4 , test tubes, Sample A= Potatoes, Sample B= Soft drink, Sample C= Onions, Sample D= Instant noodles, Sample E= Soya beans Method :

1. Two test tubes were labeled 1 and 2. 2. Tube 1 was added with 3ml of 1% albumin. 3. 3ml of KOH was added in both test tubes. 4. 2ml of CuSO4 was added slowly in both test tubes and mixed. 5. The color change was noted after 2 minutes. 6. Step 1 to 6 was repeated with 3ml of distilled water, and food sample A to E.

Part 3: Lipids Materials Brown paper, vegetable oil, water, Sample A= Potatoes, Sample B= Soft drink, Sample C= Onions, Sample D= Instant noodles, Sample E= Soya beans Method :

1. One half was written oil and the other was written water in a brown paper. 2. A tiny drop of vegetable oil was dropped at the oil and a drop of water was dropped at water. It was gently rubbed with fingertip. 3. The spots were allowed to dry. 4. The paper was hold up to the light when the spots dried. 5. Step 1 to 5 was repeated with food sample A to E. 6. Observations were recorded.

Results: Part 1: Carbohydrates Investigation 1: Monosaccharides and Disaccharides

Solution Glucose

Results Orange colour with orange precipitate (positive)

Distilled water(control) Fructose

Blue solution (negative) Orange solution with orange precipitate (positive)

Sucrose Lactose

Blue solution ( negative) Orange solution with orange precipitate (positive)

Starch Sample A

Blue solution ( negative) Orange solution with orange precipitate (positive)

Sample B Sample C Sample D Sample E

Blue solution (negative) Blue solution (negative) Blue (negative) Yellow greenish solution (positive)

Investigation 2: Starch Solution Starch Distilled water(control) Sample A Sample B 7nSample C Sample D Sample E Results Blue black solution (positive) Yellow solution (negative) Yellow solution (negative) Yellow solution (negative) Blue black solution (positive) Blue black precipitate (positive) Blue black solution (positive)

Investigation 3: Hydrolysis of Carbohydrate Tube Number Sucrose 1 Time (min) Benedicts Blue Reagent (no change) Iodine reagent Orange Blue (no change) Yellow solution with black precipitate Blue (no change) Yellow solution with black precipitate Blue (no change) Blue black 0 2 2-3 Starch 3 0 4 0 5 6-7 6 6-7 7 10 8 10

Part 2: Investigation 4: Proteins Solution Albumin Distilled water(control) Sample A Sample B Sample C Sample D Sample E Results Dark purple solution (positive) Blue solution (negative) Blue solution (negative) Purple solution (positive) Blue solution (negative) Purple solution (positive) Blue solution (negative)

Part 3: Investigation 5 :Lipids Solution Vegetable oil Distilled water Sample A Sample B Sample C Sample D Sample E Results Translucent (positive) No changes (negative) No changes (negative) No changes (negative) Translucent (positive) Translucent (positive) No changes (negative)

Discussion From the experiment, sucrose (table sugar) contains two sugars (fructose and glucose) joined by their glycosidic bond in such a way as to prevent the glucose isomerizing to aldehyde, or the fructose to alpha-hydroxy-ketone form. Sucrose is thus a non-reducing sugar which does not react with Benedict's reagent. Sucrose indirectly produces a positive result with Benedict's reagent if heated with dilute hydrochloric acid prior to the test, although after this treatment it is no longer sucrose. The acidic conditions and heat break the glycosidic bond in sucrose through hydrolysis. Amylose in starch is responsible for the formation of a deep blue color in the presence of iodine. The iodine molecule slips inside of the amylose coil.Iodine - KI Reagent: Iodine is not very soluble in water, therefore the iodine reagent is made by dissolving iodine in water in the presence of potassium iodide. This makes a linear triiodide ion complex with is soluble. The triiodide ion ion slips into the coil of the starch causing an intense blue-black color. Starches do not react or react very poorly with Benedict's reagent, due to the relatively small number of reducing sugar moieties, which occur only at the ends of carbohydrate chains.Inositol (myo-inositol) is another carbohydrate which produces a negative test. The hydrolysis of starch takes a longer time compared to the hydrolysis of sucrose because starch has a whole bunch of carbons that need to go through your hydrolysis process, where sucrose only has a few. it just takes a while to break those high number of bonds versus breaking a couple. Other than that, it is because fructose increases starch viscosity more rapidly and achieves a higher final viscosity than sucrose because fructose lowers the temperature required during gelatinizing of starch, causing a greater final viscosity. Carbohydrate can be classified into three categories which are the

monosaccharides, dissacharides and also polysaccharides.Monosaccharides are single sugar carbohydrates which is the simplest kind. In appearance, they tend to be colorless and crystalline, and dissolve easily in water. Some, but not all, are sweet-tasting. Glucose, dextrose, fructose, galactose, and ribose are all monosaccharides; so is formaldehyde. Most monosaccharides share the generic chemical formula CH2O, and the chemical structure (CHOH)nC=O(CHOH)mH. When n or m is zero, the monosaccharide is an aldose but when they are not zero, it is a ketose. Monosaccharides tend to form cyclic structures, especially in aqueous solution. This happens when a carbon links to the hydroxyl group of another carbon in a weak bond. Monosaccharides are often classified by the number of carbon atoms they contain. Monose, Diose, Triose, Tetrose, Pentose, Hexose, Heptose, Octose, and Nonose are the first nine monosaccharides. They are also classified by whether they are a ketose or aldose, and what their configuration is at the carbon-2 spot. Glucose, mannose, galactose, fructose, sedoheptulose are the exmaples of monosaccharides. Disaccharides are

carbohydrates that are created when two monosaccharides are joined. The creation of a disaccharide involves the union of two monosaccharides that undergo a process in which a water molecule is removed as part of the union. Like the two monosaccharides that combine to form the single disaccharide, the carbohydrate is sweet to the taste and tends to dissolve in water with relative ease. Maltose, cellobiose, lactose, sucrose are the examples of dissacharides. Maltose is made up of 2 glucose molecules. Lactose is made up of galactose and glucose. Sucrose is made up of glucose and fructose. Polysaccharides are the complex carbohydrates. They are made up of chains of monosaccharides (the sugars) which are linked together by glycosidic bonds, which are formed by the condensation reaction. The linkage of monosaccharides into chains creates chains of greatly varying length, ranging from chains of just two monosaccharides, which makes a disaccharide to the polysaccharides which consisted of many thousands of sugars. Starch, glycogen and dextran are the examples of polysaccharides. Starch is a mixture of polymeric glucose which are the amylose and amylopectin. Benedicts reagent test is the most common test which indicates the presence of most small sugars. Benedict's solution contains copper sulfate that reacts with sugar to form copper oxide, a reddish brown powder. The more red the solution looks, the more copper oxide there is, and therefore the more sugar is in the solution. Reducing sugar is oxidized when heated with Benedict's solution, which means the sugar looses electrons. Benedict's solution contains copper sulphate. The copper 2+ ions in copper sulfate gain an electron that the sugar has lost

and become copper 1+ ions - this is called reduction. The copper ions immediately react with oxygen to form copper oxide which forms a reddish brown precipitate. The more red the solution looks, the more copper oxide there is, and therefore the more sugar is in the solution. Iodine solution is used to indicate the presence of starch. Starch is a coiled polymer of glucose. Iodine reacts with the coiled molecules and turns bluish-black when added to a solution. A solution that remains a yellowish-brown color is a negative test for starch, whereas one that turns bluish-black is a positive test for starch. Hydrolysis chemical reaction in which water reacts with a compound to produce other compounds. It involves the splitting of a bond and the addition of the hydrogen cation and the hydroxide anion from the water. Below is one of the example of hydrolysis process which occurs in carbohydrate.

If the light blue solution changed to an orange-brown solution in a boiling water bath, that means, its a positeve Benedicts reagent test. This indicating the presence of glucose in the solution. Sucrose is a non-reducing sugar which does not react with Benedict's reagent. But, sucrose produced a positive result with Benedict's reagent after the solution heated with dilute hydrochloric acid prior to the test. The acidic conditions plus the heat given is able to break the glycosidic bond in sucrose through hydrolysis process. Iodine is used to indicate the presence of starch. This is a simple test where a few drops of iodine is added to a given solution. Iodine is added to test-tube 4, 6, and 8. Only the solution in test tube 8 turned blackblue which means there is a presence of starch. The solution in test tube 4 and 6 remained the color of iodine, reddish-orange, which means there is no starch present, a negative test. Starch didnt react with Benedict's reagent in test tube 3, 5 and 7 due to the relatively small number of reducing sugar which occur only at the ends of carbohydrate chains.

The hydrolysis of starch is longer than the hydrolysis of sucrose. This is because starch is a very long chain of polysaccharide which has a whole bunch of carbons that need to go through the hydrolysis process, where sucrose only has a few. It just takes short time to break those high number of bonds versus breaking a couple. Besides that, starch also has a high viscosity compared to sucrose due to the presence of fructose in starch.

References 1. http://www.wisegeek.com/topics/monosaccharides.htm# 2. http://www.suite101.com/content/polysaccharides-a28515 3. http://au.answers.yahoo.com/question/index?qid=20071022112730AA58yJU 4. http://www.scientificpsychic.com/fitness/carbohydrates.html 5. http://www2.chemistry.msu.edu/faculty/reusch/VirtTxtJml/carbhyd.htmhttp://ww w.biology-online.org/dictionary/Monosaccharide 6. http://biology.unm.edu/ccouncil/Biology_124/Summaries/Macromol.html 7. http://terpconnect.umd.edu/~nsw/ench485/lab5.htm 8. http://www.elmhurst.edu/~chm/vchembook/548starchiodine.html 9. http://www.docstoc.com/docs/26034865/Hydrolysis-of-Starch