Identifying Organic Compounds

 

Introduction: (great tutorial: http://www.occc.edu/biologylabs/Documents/Organic%20Compounds/Organic%20Compounds.htm )

 

The most common organic compounds found in living organisms are lipids, carbohydrates, proteins, and nucleic acids.  Common foods, which often consist of plant materials or substances derived from animals, are also combinations of these organic compounds.  Substance called indicators can be used to test for the presence of organic compounds.  An indicator is a substance that changes color in the presence of a particular compound.  In this investigation, you will use several indicators to test for the presence of lipids, carbohydrates, and proteins in various foods.

 

Problem:  What are the major types of organic compounds in some common foods?

 

Pre-lab Discussion:

 

  1. What is an indicator?  How are indicators used in this experiment?
  2. What is the purpose of using distilled water as one of your test substances?
  3. What is the controlled variable for each group of testing?
  4. What is the purpose of washing the equipment thoroughly?
  5. What is the IV? DV?
  6. You have added Sudan IV stain to each substance.  What change indicates the presence of lipids?

 

Substances being tested:

 

  1. honey solution
  2. egg white solution
  3. corn oil
  4. lettuce solution
  5. gelatin solution
  6. yolk solution
  7. potato solution
  8. apple juice
  9. distilled water

 

Part A:  Testing for Carbohydrates: Monosaccharides

 

Glucose, commonly called “blood sugar,” and fructose, “fruit sugar,” are two of the most common monosaccharides.  Ribose, an important component of nucleic acids, is also a monosaccharide and will be discussed during our study of DNA and RNA.  Monosaccharides can be identified in a substance using an indicator called Benedict’s solution.  When Benedict’s is heated in the presence of a monosaccharide, the color changes from blue to green to yellow to reddish-orange, depending on the amount of monosaccharide present.

 

 Procedure:  You can start step 1. and then proceed to another test while the water heats up.

  1. Turn on the hot plate to high and fill a beaker half full of tap water.  Place the beaker on the hot plate.  While this is heating, add 5mL of each sample to a test tube.  Add 10 drops of Benedict’s solution to each test tube. 
  2. Place the test tubes in the hot-water bath.  Heat the test tubes for 3 to 5 minutes, or until the indicator changes color, if it changes color. 
  3. With a test tube holder, remove the tubes from the hot-water bath and place them back in the test tube rack. 
  4. Record any changes in your data table.

 

Part B: Testing for Carbohydrates: Polysaccharides

 

Monosaccharides may join together to form long chains called polysaccharides that may be either straight or branched.  Starch is an example of a polysaccharide formed entirely of glucose monosaccharides.  Polysaccharides can be tested for using the indicator Lugol’s reagent (iodine/potassium iodine).  Lugol’s will change color from yellow-brown to blue-black-purple in the presence of starch.

 

Procedure:

  1. Add one dropper full of each solution to a well in the well tray. 
  2. Add 2-3 drops of Lugol’s to each sample. 
  3. Record any changes in your data table.

 

 

Part C: Testing for Lipids

 

Lipids can be generally categorized into two subgroups: saturated fats and unsaturated fats.  In saturated fats, the carbon chains are bonded together with only single covalent bonds and the remaining electrons are bonded to hydrogen.  This structure keeps the hydrocarbon tails relatively straight, so they readily align with each other, pack closely together, and thus tend to be solid at room temperature.  Saturated fats are most common in animals.  Unsaturated fats has double bonds between the carbons and therefore do not have the maximum number of links to hydrogen.  These regions of double bonding allow kinks to form in the hydrocarbon tails, keeping them from packing closely together.  For this reason, unsaturated fats are usually liquid at room temperature.  Unsaturated fats are common in plants.  Because fats are nonpolar, we will test for their presence using Sudan IV, a nonpolar dye that will readily mix with lipids but form small bubbles or separate with non-fats.  Lipids can also be tested for using the “sack lunch” method: the lipid is rubbed into a brown bag.  If a grease spot appears (the brown bag appears translucent), then a lipid is present in the food.

 

Procedure: Sudan IV

  1. Add one dropper full of each solution to a clean well in the well tray.
  2. Add 3-5 drops of Sudan IV to each sample.
  3. Look for small bubbles or separation.
  4. Record results in data table.

 

Procedure: Sack lunch

  1. Add one dropper full of each solution to a small piece of brown bag.
  2. Rub the solution until a “wet” spot appears on the paper.  Rub off any excess with a paper towel.
  3. Set the paper aside until the spot appears to dry, about 10-15 minutes.
  4. Hold the brown paper up to a bright light.
  5. Record observations in data table.

 

Part D: Testing for Proteins

 

Proteins are made up of one or more polypeptides, which are linear polymers of monomers called amino acids.  Amino acids derive their name from the amino group and the carboxyl group (which is acidic).  Polypeptides are formed when amino acids are joined together by peptide bonds between the amino group of one amino acid and the carboxyl group of another amino acid.  Proteins can be tested for using Biuret’s reagent, which reacts with the peptide bond between amino acids in the polypeptide.  Biuret’s reagent turns from blue to purple-violet if peptides bonds are present.

 

 

 

Procedure:

 

  1. Add one dropper full of each solution to a clean well in the well tray.
  2. Add 3-5 drops of Biuret’s reagent to each sample.
  3. Record any changes in color.

 

 

Rubric for Identifying Organic Molecules  (24 points)

Title: Must be descriptive (includes the IV and DV of the experiment) 1 point

Results:

Data Table of class results:  (8 points)

a. includes a descriptive title (remember you can most likely use the regular title again).1 point

b. only one horizontal line between the column headings and the data  (column headings include the name of the indicator and what the indicator detects in parenthesis... example: Benedict's Solution (monosaccharides)... (5 points)

c. Key for explaining the symbols you might use... example "+" = positive; also and colors or changes that a positive test indicates (example: Benedict's solution positive test includes a green, red, orange, or yellow solid) (2 points)

Post Lab Questions  make sure you pick one version of questions to answer and provide any citations requested for full credit. Also remember incomplete sentences, sentences starting with "it" or "because", or sentences using pronouns will not receive credit and you will be asked to redo the assignment.

Answer questions  1-5 to earn a C on this section: (5 points), answer questions 6-9 to earn a B (8 points), to earn an A (up to 11 points answer 6-10).

  1. Why is a monosaccharide called a monomer?
  2. Why is a polysaccharide called a polymer?
  3. List which chemical detects which molecule.
  4. A lipid refers to three different types of substances, a _____________, a _____________ and __________.
  5. Why is water a good substance for a control in this lab activity?
  6. People with diabetes are instructed to avoid foods that are rich in carbohydrates.  How could your observations in this investigation help you decide whether a food should be served to a person with diabetes?  ( explain the relationship between diabetes and monosaccharides and there connection to this lab activity... include citations to references) (2 points)
  7. What conclusion could you make if a positive test for any of the organic compounds occurred in the distilled water?  (what is in distilled water normally... what could the source of this "positive" test come from during the lab testing process). (2 points)
  8. Why can’t Biuret’s reagent be used to test for amino acids? (what does Biuret's actually detect on a  protein, what is the relationship between an amino acid and a protein? cite your sources at the end of your answer..) (2 points)
  9. The leaves of many plants are coated with a waxy substance that causes them to shed water.  How would you expect this substance to react in the Sudan IV test?  ( what type of biomolecule is detected by Sudan's what is the relationship to wax... what is the name of the waxy substance... cite your sources at the end of your answer.) (2 points)
  10. In winter, plants exchange the saturated lipids in their cell membranes for unsaturated lipids.  Unsaturated lipids are “bent” and keep the membranes more fluid, or pliable, because they cannot be stacked closely together.  Of what advantage would this be for green stemmed (herbaceous) plants that live through the winter?  (Hint: what happens to bacon grease or the grease on top of chicken soup when you put it in the refrigerator?) ( Which is a liquid at room temperature, a solid at room temperature.. why would a plant want to switch the types of fatty acids in the winter... what could happen if the cell membrane only had saturated fatty acids during the winter, or only unsaturated fatty acids in the summer... don't forget to cite your sources properly at the end of your answer.)_(3 points)

Helpful articles to answer #10 article #1  http://tea.armadaproject.org/atwood/12.2.1998.html  (Especially see section F)

Use correct format (see http://www.srvhs.org/staff/teachers/rgroch/bio/bio_home_page/biolabinstr.htm  see Section 8 for help).