Name: Period: Date: Testing for Biological Macromolecules Lab
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1 Testing for Biological Macromolecules Lab Introduction: All living organisms are composed of various types of organic molecules, such as carbohydrates, starches, proteins, lipids and nucleic acids. These substances, contained in the foods we eat, provide us with the energy and cellular building blocks necessary for life. Several of these molecules can be detected by simple chemical tests. In today s lab, you will perform tests to determine carbohydrates, lipids and proteins, and apply these tests to unknowns. Carbohydrates Background: Sugars are small carbohydrate molecules used as a source of energy by all organisms and manufactured in the chloroplasts of plants. There are many kinds of carbohydrate molecules, all of which are made up of various arrangements of carbon, hydrogen, and oxygen atoms. The simplest carbohydrates are monosaccharides, or simple sugars, such as glucose and fructose. Disaccharides consist of two bonded monosaccharides. Examples of disaccharides include lactose, the sugar contained in milk; sucrose, common table sugar; and maltose. Polysaccharides are large macromolecules comprised of hundreds or thousands of linked monosaccharides. Some of these polysaccharides are broken down as needed for energy. A common polysaccharide, starch, is a storage polysaccharide found in plants. Other polysaccharides serve a structural function, such as cellulose, a polysaccharide that is the main component in plant cell walls and is the most abundant organic compound on earth. Testing for Monosaccharides (Simple Sugars): To test for the presence of simple sugars (monosaccharides) you will use a solution known as Benedict s Reagent. Benedict s reagent is a clear blue solution due to the cupric copper ion (Cu ++ ) present, but when combined and heated to boiling with a substance containing a monosaccharide, the cupric ions are reduced to insoluble red cuprous ions (Cu + ). Therefore, a positive reaction in a Benedict s solution test will change the clear light blue solution to a green and then to an orange-brown color. Disaccharides do not react with the Benedict s reagent. 1. Prepare the test tubes according to Table Place all test tubes in a boiling water bath for approximately 3-5 minutes 3. Remove all test tubes and record results Table 1 Test Solutions Benedict s Reagent 1. Water 5ml 5 drops 2. 1% Glucose 5ml 5 drops 3. 1% Lactose 5 ml 5 drops 4. 1% Starch 5ml 5 drops Color of solution Before heating After heating Presence of monosaccharides?
2 Testing for Polysaccharides: To test for the presence of starch you will use a solution known as Iodine (IKI). If a polysaccharide such as starch is present in the presence of iodine, the iodine will lodge itself in the polysaccharide chain and give it a black-blue color. Therefore, if solution turns black-blue when iodine is added to it, then starch is present. If the solution remains the color of iodine, reddish-orange, there is no starch. 1. Prepare the well plates according to Table Record results Table 2 Test Solutions Iodine Color of solution Before adding iodine After adding iodine 1. Water 5 ml 5 drops 2. 1% Glucose 5 ml 5 drops 3. 1% Lactose 5 ml 5 drops 4. 1% Starch 5 ml 5 drops Presence of polysaccharide? Testing Carbohydrate Breakdown: To test for the breakdown of carbohydrates, you need to use an enzyme called amylase. Amylase is an enzyme found in your saliva that helps to break down polymers into monosaccharides. You are to perform the following procure to observe this process. 1. Put 6 ml of starch solution into test tube A 2. Add about 3 ml of amylase (saliva) to test tube A 3. Cover the open end of the test tube with parafilm and shake test tube 5 times to mix the substances BEGIN TIMING IMMEDIATELY AFTER STEP 3 AND EVERY 2 MINUTES YOU WILL. 4. Take small sample from test tube A and add to a clean test tube 5. Add 4 drops of Benedicts solution to the test tube with the small sample 6. Heat the test tube in a water bath for 1minute 7. Record color change 8. Clean the used test tube to be used again. 9. Repeat steps 4-8 every 2 minutes until you reach 14 minutes. Time Color Starch or simple sugar present
3 Testing For Proteins The Biuret Test is often used to determine the presence of the peptide bonds in protein. Peptide bonds are the bonds that are formed between amino acids (the monomers of proteins). 1. Add 3 ml of the sample A to a clean, dry test tube. 2. Add 2 ml of Biuret Reagent. 3. Shake well 4. Repeat steps 1-4 with sample B and C. 5. Allow the mixtures to stand for 5 minutes. 6. Observe any color change. Color of original Biuret Solution: Substance Color After Adding Biuret Test Result ( + or - ) For Protein A B C What substance was the protein solution? How do you know?
4 Testing Unknowns: Use the tests that you have just learned and mastered to determine the biological molecules present in the following unknowns A, B, C, D and E. You will determine which unknown is a monosaccharide, which is a disaccharide, which is a polysaccharide and which is water. You will complete the data table below as you complete your testing. Sample Macromolecule present Rationale A B C D E Additional Questions to be answered. 1. What is the role of water in this activity? 2. Summarize what you learned about testing for starch and simple sugars.
5 3. Explain the relationship between starches and simple sugars and the role of the amylase when added to starches. 4. What did you learn about testing for proteins?
For example, monosaccharides such as glucose are polar and soluble in water, whereas lipids are nonpolar and insoluble in water.
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