Identifying Biological Molecules with Colorimetric Chemical Tests
Objectives
Perform a series of accurate tests on biological molecules to detect the presence of carbohydrates and proteins, as well as the action of an enzyme on specific molecules.
Introduction
Unknown biological molecules can be correctly identified using various tests, including colorimetric chemical tests. Colorimetric tests are when a chemical reagent, such as Benedict’s, is used on a substance or solution and a certain color change occurs as a response. Carbohydrates and proteins, two of the larger types of biological molecules, also called macromolecules, are actually chains of molecules called polymers. The smaller molecules that build up these chains are
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The results for Benedict’s test for reducing sugars before hydrolysis, the control dH2O had no color change, as well as sucrose and raffinose. Gelatin became a dark blue and egg albumin a light greyish blue. Glucose yielded a dark orange, milk albumin turned orange, and starch had a yellow precipitate. In the Benedict’s test for reducing sugars by hot acid hydrolysis, the control dH2O was blue or no reaction. Glucose turned a brownish-orange; sucrose, a reddish-brown, raffinose, light pink; starch, a dark yellowish-orange; gelatin, violet; milk powder, a light yellow; and egg albumin, a greyish-violet. In the Lugol’s test for polysaccharides, the control dH2O turned yellow or no reaction, as well as glucose, sucrose, raffinose, milk powder and albumin. Starch was clear with blue-violet on the bottom (precipitate) and gelatin turned a slightly opaque white. In the test for polysaccharides remaining after hydrolysis with Lugol’s solution the control dH2O was yellow or no reaction. Glucose, sucrose, raffinose, starch and gelatin also had no reaction. Milk powder had a white precipitate and particle suspension. Egg albumin had an even greater white precipitate and particle suspension. In Biuret’s test for proteins there was no reaction for dH2O, glucose, sucrose, raffinose, and starch. Gelatin, milk powder and egg albumin all turned slightly violet with bubbles. In part 2 of the lab, we tested the effect of the enzyme invertase on dH2O (the control), sucrose, and
4. There are other types of reagents used to determine what type of biomolecule a substance is. For example, copper ions present in Benedict’s reagent reacts with the free end of any reducing sugars, such as glucose, when heated. Originally blue in color, these copper ions are reduced by the sugar, and produce an orange-red colored precipitate. Alternatively, iodine-potassium iodide (IKI) may also be used when working with starch. IKI contains special tri-iodine ions which interact with the coiled structure of a starch
1. Benedict’s solution is added to white grape juice and heated. The color changes from blue to orange. Based on this result, what biological molecules are
The purpose of this lab was to test different substances using various procedures to see what biomolecules were present and ultimately find out what restaurant Anna Lyza had eaten at before she died. For the first control test, we used vegetable oil to test for lipids. So, if the solution does not contain lipids, it does not become translucent when placed onto a paper bag square and held up to a light. So, it is a negative result. However, in the presence of lipids, the solution will become translucent when placed onto a paper bag square and held up to a light. Therefore in this case, the result is positive. On the other hand, we used albumin egg to test for proteins in another control test. If the solution does not contain proteins, it will not experience any color change and so it is a negative result. When there are proteins existing in the solution, it will turn bluish/purplish and for this reason it is a positive result. Furthermore in the third control test, we used dextrose to test for simple carbohydrates such as glucose. If the solution does not contain simple carbohydrates, it will not undergo any color change and will remain a blue color. So, it is a negative result in this circumstance. If there are simple carbohydrates present in the solution, the solution will turn reddish and so the result is positive. For the last control test, we used starch solution to test
All living things contain some form of organic macromolecules including: Lipids, proteins, carbohydrates and nucleic acids. All of these organic molecules are alike in the sense that is they are made up of bonded elements such as carbon, hydrogen, oxygen, and to smaller quantities nitrogen, phosphorus and sulfur. The macromolecules each contain large long chains of carbon and hydrogen atoms and often consists of repeating smaller molecules bonded together in a repeating pattern (polymers). To test whether a specific solid white substance is a protein is simple due to the unique chains found in the respective type of macromolecule. To test if the substance is a carbohydrate, two tests could be performed on separate samples of the object. First,
The use of multiple test tubes and Parafilm was used for each experiment. Catechol, potato juice, pH 7 phosphate buffer, and stock potato extract 1:1 will be used to conduct the following experiments: temperature effect on enzyme activity, the effect of pH on enzyme action, the effect of enzyme concentration, and the effect of substrate concentration on enzyme activity. For the temperature effect on enzyme activity, three test tube were filled with three ml of pH 7 phosphate buffer and each test tube was labels 1.5 degrees Celsius, 20 °C, and 60 °C. The first test tube was placed in an ice-water bath, the second test tube was left at room temperature, and the third test tube was placed in approximately 60°C of warm water. After filling the test tubes with three ml of the
The most common macromolecules found in living organisms are lipids, carbohydrates, proteins and nucleic acids. Briefly, the meaning of macromolecules is that they normally contain two or more molecules in them and their main functions are to store energy, information and much more. Most foods are known to be combinations of macromolecules. While some of these compounds can be detected by taste tests, many cannot. Scientists then use certain tests to determine the presence of macromolecules.
A cell, the building block of all living organisms, is composed of four fundamental biomolecules: proteins, carbohydrates, sugars and lipids. Proteins provide a vast amount of functions cells such as they serve as enzymes, provide structural support to cells, and act as antibodies. Reagents are used to spark a chemical reaction. The reagent used to detect protein traces in a substance is Biuret’s. Biuret’s will turn purple if proteins are present and blue if they are none. Biuret’s copper particles, have a charge of +2, are diminished to a charge of +1 when peptide bonds, which are in proteins, are present, creating the color change. Polysaccharides, which are carbohydrates, are most notably known to provide energy to the body, but they also help in breaking down fatty acids. Iodine is the reagent used to determine whether a substance has starch in it. The iodine/starch complex has energy levels that are only for retaining unmistakable light, giving the complex its extraordinarily dark black-blue shade. If there is no starch found, iodine will remain its natural yellowish-brownish color, but if starch is present, iodine will turn blue-black. Monosaccharides, which are sugars, like polysaccharides, provide the body with energy. To detect monosaccharides, the reagent, Benedict’s, is used. Benedict’s reagent is added to a test tube, then it is placed in
3. State the name and structure of the functional group for each type of biologically
b) Benedict test the solution color will change from blue to pink/orange red, indicating simple sugars are present. Lugols test the solution color will change from yellowish brown to dark purple, indicating starch and polysaccharides are present. Sudan iV test the lipid content will turn into red, indicating lipids are
In this lab experiment the action of the enzyme Amylase was observed on starch (the substrate). Amylase changed the starch into a simpler form, the sugar maltose, which is soluble in water. Maltose then breaks down the glucose chains of starch in the pancreas and intestines. Amylase is present in human saliva, and begins to act on the starch in the food while still in the mouth. Exposure to heat or extreme PH (acid or base) will denature proteins. Enzymes, including amylase, are proteins; if denatured enzymes can no longer act as a catalyst for the reaction. In the presence of potassium iodide, starch turns a dark purple color; however maltose does not react with I2KI. The rate of fading of starch allows a quantitative measurement of reaction rate.
enzymes that will be used during this lab to test the ability of amylase to break down starch ,a
In part II of the lab six small glass tubes were obtained in a test tube rack. Ten drops of distilled water were then added to test tube 1, five drops to tubes 2-4, and no drops in tubes 5 and 6. Five drops of 0.1M HCl were added to test tube 5 and five drops of 0.1M NaOH to test tube 6. Five drops of enzyme were then added to all tubes except tube 1. Tube 3 was then placed in the ice bucket and tube 4 was placed in the hot bucket at 80-900C for five minutes, the remaining tubes were left in the test tube rack. After the five minutes five drops of 1% starch was added to every tube and left to sit for ten minutes. After ten minutes five drops of DNSA were then added to all the tubes. All the tubes were then taken and placed in the
The guiding question of this ADI lab was, “What are the identities of the unknown compounds?” The goal of this lab was to understand the relationships between moles and molar mass to find the identity of unknown compounds. The mole can be used to measure small amounts of a substance or is used to convert from unit to unit using dimensional analysis. One mole is equivalent to the molar mass in grams of that substance. If you start with the moles of an unknown substance, multiply it by a given compound’s molar mass, and then divide it by however many moles are in the compound of your choice, you will get the mass of the compound. With that answer you can then compare with mass of the compound in the bag to determine its identity. We first started
This experiment consisted of setting up a control group of starch in various temperature and then placing both fungal amylases and bacterial amylases in a mixture of starch and placing the solution of amylase and starch in various temperatures of water. After a certain amount of time- different amount of time needs to be used in order to have reliable results- iodine is added in a well on spot plates, then two drops of the mixture of amylase-starch is added from each temperature used, by adding iodine into the plates the mixture will show how much starch was hydrolyzed, this is used to calculate the amount of
B) Proteins Proteins are made up of chains of amino acids. Each protein folds into a characteristic three dimensional shape that is essential to its function. They perform a variety of functions in cells and are probably the most diverse macromolecule in living systems. The Biuret test detects the presence of proteins and short peptides (short chains of amino acids). Biuret reagent contains a strong solution of sodium hydroxide (NaOH) and a very small amount of dilute copper sulfate solution. The reagent changes color in the presence of a protein because the amino groups in the amino acids react with the copper ions producing a violet color. A negative result is blue.