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Daiva Kuncaite Bio 201 Prof. Wijesinghe 09/29/2009 Week II Title: Organic Molecules Objectives: carry out the experiment to determine if the substance is organic or inorganic. Use various regents and methods to identify organic substances class. Identify organic compounds found in the assigned unknown solution. Introduction: The key factor that distinguishes organic substances from inorganic is the element carbon. The main organic compound find in living organisms are protein, lipids, carbohydrates and nucleic acids. Carbohydrates ratio of C:H:O is 1:2:1. They are polar molecules and soluble in water. The simplest carbohydrates are monosaccharide (ex. Glucose). Two monosaccharide forms disaccharides (ex. Sucrose, lactose) and more than two molecules of monosaccharide form polysaccharide (ex. Starch). Lipids in their structure contains less O than in carbohydrates, therefore lipids are nonpolar and water insoluble. Proteins are polymers of the amino acids. Proteins have complex structure but also have a vast variety of functions. Procedure: Since we carried many experiments I will explain each experiment procedure individually. To determine if the substance is organic we took a small amount of each desired substance in the test tube and using a Bunsen burner checked if the substance burned and blackened. Each test tube we hold using test tube holder and away from the face in order to avoid any damage. To carry on further experiments we had always prepare positive control and negative control test tubes. The negative control always indicated that molecules we are looking for are not presented. The positive control always indicates the presence of desired molecule. For each experiment water served as negative control. However, positive control changed accordingly to the experiment. To determine presence of reducing sugar, which is mainly mosaccharides, we took each desired substance about 2 ml in the test tubes and added 2-3 drops of Benedict’s reagent. Then we brought the water to boiling in the beaker on the heating plate. Placed each test tube in the hot water bath for no longer than one minute and observed the changes. The positive control was glucose solution. To determine the presence of starch we used iodine reagent. We pour about 2 ml of each desired substance in the test tube and added one drop of iodine in each. We observed the appearance of the blue-black color. The positive control was starch solution. To determine the presence of fats we did test using Sudan IV reagent, which stains the fats. We took about 5 ml of desired substance in the test tubes and added 5 drops of the Sudan IV reagent, which stained the fats red. The positive control was vegetable oil. To determine the presence of fats we also used Greasy Spot Test. On the white paper towel we lay out the desired substances, crush them, and labeled them. After the substances dried we recorded our observations. To determine the presence of proteins we used Biuret reagent. We took 2 ml of the desired substances in the test tube and added 2-3 drops of Biuret reagent and recorded the appearance of purple or lavender color which indicated the presence of protein. The positive control was Albumin solution. The professor assigned the unknown solution # 2 to test for the Daiva Kuncaite Bio 201 Prof. Wijesinghe 09/29/2009 Week II presence of any organic substance(s). We divided assigned solution in 4 test tubes. First test tube we test for reducing sugar by adding few drops of Benedict’s reagent’s in the solution and placing the test tube in the hot water bath. The second test tube we tested for starch by adding few drops iodine. The third test tube we tested presence of protein by adding Biuret reagent. The last test tube we test for lipids by adding few drops of Sudan IV reagent. Results and Conclusions: Determination if the substance is organic Since gelatin, sugar and milk had burned and blackened we concluded that these three substances are organic, whereas salt and chalk dust are inorganic compound and didn’t changed color during the experiment. Reducing Sugar The negative control test tube with water after adding Benedict’s reagent turned sky blue and remain the same after the hot bath, whereas the positive control test tube with the glucose solution turned orange after the hot water bath. Therefore the color change to yellow/orange indicated the presence of reducing sugar. The positive results appeared in milk, Apple juice, dilute molasses, onion juice, lactose and sucrose solution. Sucrose solution supposed to give negative results if we use pure sucrose. However, the industrial sucrose we used in the experiment might have contained glucose contaminations which lead to the positive results of the test. Since lactose gave positive result, milk also showed the positive results sign in yellowish appearance, because it contains lactose as component. Therefore lactose is a reducing sugar. Powder cracker in water and starch solution showed no presence of the reducing sugar as it was expected. Starch The negative control test tube after adding iodine turned amber-brown color, which indicated no presence of starch. The positive control test tube with starch solution had turned black-blue color after adding iodine. In addition, we received positive results indicating the presence of starch in paper, potato, and bread. However, paper contains cellulose, a different type of polysaccharide then starch, therefore we got positive results. Fat test using Sudan IV The negative control test tube with water after adding reagent turned pink, evenly distributed color. The positive control test tube was mixing water with oil and few drops of reagent, which colored in red the fat layer on the surface of water. However, when we added the reagent in the Daiva Kuncaite Bio 201 Prof. Wijesinghe 09/29/2009 Week II third test tube which contained water, oil and detergent we observed emulsification. The red droplets of oil appeared instead of layer and the boundaries between oil and water diminished. Greasy Spot Test Water stain as negative control had dried without leaving oily stain on the paper towel. The vegetable oil, a positive control had left an oily stain after drying. In addition potato chip and peanuts gave positive results which indicated the presence of fats. Sucrose solution gave negative result as it was expected. Protein The negative control test tube containing water gave a blue color after adding Biuret reagent. The positive control test tube containing Albumin, which is a protein, after adding reagent, gave purple color. Milk and crushed peanuts showed slightly purple color, which indicated that they have some amount of protein present. However, sucrose solution, powdered cracker and vegetable oil remained blue colored, which indicated no presence of the proteins. Unknown #2 The test for starch and protein gave positive results, which lead us to the conclusion that the unknown #2 contains two organic substances: starch and protein. Test for reducing sugar and lipids gave negative results. Homework: 1. What reagent do you use to test for a monosaccharide? Benedict’s reagent. 2. Do all of the tests require heating? Which one do? Not all tests require heating. Only test for reducing sugar using Benedict’s reagent and in determination if the substance is organic. 3. What color is a positive test for protein? Purple/lavender 4. Why does sucrose (table sugar) give a negative result for Benedict’s test? Sucrose is non reducing sugar and doesn’t react with the Benedict’s reagent. However, the industrial sucrose we used in the experiment might have contained glucose contaminations which lead to the positive results of the test. Daiva Kuncaite Bio 201 Prof. Wijesinghe 09/29/2009 Week II 5. What are the monomers of proteins called? Amino acids. 6. Most lipids fall into the category of triglycerides. What is the term for a triglyceride that is a liquid at room temperature? Unsaturated A solid? Saturated 7. What simple test can you run to tell if a substance is organic or not? Burn the material. If the material burns and blackens the material is organic. 8. What element do all organic substances contain? Carbon