Lab 1: Enzymatic Reactions

Teacher Preparation:

• If possible, it is recommended that both Parts A and B of this lab be completed in one class period.
• Review information found in the Background Agricultural Connections section of the lesson, lesson Procedures, and the attached Essential Files.
• Prepare the following for each group:
  • Part A: Cut thin slices for students to taste and place them on paper plates. In addition, slice apples, oranges, and bananas for the investigation in half. Each group will be given a 1/4 piece of each fruit type. If possible, this should be started at the beginning of the lesson so your students can observe the process.
    • Tip: The extra apples are needed for the second lab in this lesson. It is important these apples remain in whole form until you are ready to begin the second lab to prevent enzymatic browning from occurring. Additionally, classroom temperature can affect how quickly the apple will brown. If your classroom is generally cold, oxidation effects may be less obvious. You may need to increase the amount of time students wait prior to observing a large change in the color of the apple’s flesh.
    • Timesaver: Prepare fruit for the students ahead of time. The apples will be the only exception, as they should brown quickly. For quicker preparation of apples, use an apple corer. You may also consider assigning fewer fruits to each group and allow data to be shared. However, be sure to provide each group with one oxidizing fruit and one non-oxidizing fruit (e.g. apple and orange, or banana and orange). Be sure to adjust materials needed for this adaptation.
  • Part B:
    • Students will choose 2 treatments (e.g. vitamin C, baking soda, etc.) to include in their investigation. Organize materials in one location for easy distribution. Students will prepare their own solutions (if applicable). Have these substances available at different areas/stations in the room to maintain student flow and prevent crowding.
      • Tip: If desired, some student groups may need to be asked to test certain substances to be sure all substances are tested.
      • Timesaver: Prepare solutions ahead of time. Keep in mind each group will need to be able to test their prediction. This may require students to complete their predictions prior to the planned lab or the preparation of multiple samples for each possible substance.

Procedures:

1. Distribute lab materials. It is recommended that materials are organized into stations for easier distribution. Students should be arranged in small groups of 4-5. Each group should receive the lab supplies outlined in the Materials section as well as 1 copy of the Enzymatic Reactions lab sheet and the Oxidation Transformation handout.
2. Before beginning the lab investigation:
   • Ask students to read Oxidation Transformation and complete the focus questions for this lab investigation.
   • Require students to wash their hands.
   • Allow students to taste a sample of apple, orange, and banana prior to beginning any investigation procedures. This process is important for increasing student exposure to healthy foods and decreasing the likelihood that students will be tempted to taste foods included as investigation materials.
   • Emphasize the importance of practicing good food safety behaviors by not consuming substances during the lab investigation.
3. Show the Enzymatic Reactions Part A video to demonstrate the lab.
4. Launch Part A of the lab by asking students to prepare their fruit. Students should then observe and make a prediction about their unknown substances. Instruct students to pay special attention to the timer as they collect their data. Students should see the following:
   • Apple: Students should observe the apple’s flesh turning a brown color. As time passes, the brown color will become more distinct.
   • Orange: Students will not observe any browning on the flesh of the orange.
   • Banana: Students should observe browning of the banana slices. Browning will likely be less intense compared to the apples. The peel of the banana will also brown.
5. Allow students to work in small groups on the Enzymatic Reactions lab sheet to further explore the topic and respond to lab questions.
6. After completing the observation of enzymatic browning among three types of fruit and the investigation’s conclusions, students should be prepared to begin Part B. Be sure student groups set aside the apple sample from Part A of the lab investigation to serve as the control in Part B. If completing the lab over two class periods, store apple slices in sealed plastic bags.
7. Distribute student lab materials for Part B with students organized into stations for easier distribution. Students should continue in groups of 4-5.
8. Watch the Enzymatic Reactions Part B demonstration video.
9. As a group, students should choose two substances they predict would prevent enzymatic oxidation.
10. Students will prepare the chosen substances and test their prediction using an apple.
11. Allow students to work in small groups on the Enzymatic Reactions lab sheet to further explore the topic and respond to lab questions.
12. Use the following information for a breakdown of expected reactions:
    • Vitamin C: Little to no browning should occur due to the presence of vitamin C (antioxidant).
    • Cream of Tartar: Little to no browning should occur. Cream of Tartar is an acid and will lower the pH of the fruit’s flesh.
    • Lemon Juice: Little to no browning should occur due to the presence of vitamin C (antioxidant).
    • Sugar-water: Some browning should be expected. Sugar water may coat the surface of the fruit’s flesh to prevent extensive browning.
    • Baking Soda: Browning should be expected. Baking soda will increase the pH of the fruit’s flesh and hasten the browning effect.
    • Vinegar: Little to no browning should occur. Vinegar is an acid and will lower the pH of the fruit’s flesh.
    • Salt Water: Some browning should be expected. Salt water may coat the surface of the fruit’s flesh to prevent extensive browning.
    • Water: Water can temporarily prevent browning if the fruit is submerged. It works by decreasing exposure of the fruit’s flesh to oxygen. If fruit is simply dipped in water, oxidation will occur. This sample can be used as an investigation control.

Lab 2: Hidden Antioxidants

Teacher Preparation:

1. Review information found in the Background Agricultural Connections section of the lesson, lesson Procedures, and the attached Essential Files.
2. Prepare materials for each group. Place approximately 20 drops of iodine into each 2 oz. or similar size cup. Set up stations from which students can obtain the test substance chosen.
   • Tip: Groups may be assigned their test substance if it is desired that all substances be tested.
   • Timesaver: Prepare solutions ahead of time. Keep in mind that each group will need to be able to test their prediction. This may require students to complete their predictions prior to the planned lab or the preparation of multiple samples for each possible substance.
   • Tip: If sufficient amount of materials are not available, the testing of the substances may be done as a demonstration.

Procedures:

1. Before beginning the lab investigation:
   • Be sure students read Oxidation Transformation and completed the focus questions in lab one.
   • Require students to wash their hands.
   • Allow students to taste a sample of the food highlighted in the lesson prior to beginning any investigation procedures. This process is important for increasing student exposure to healthy foods and decreasing the likelihood that students will be tempted to taste foods included as investigation materials.
   • Emphasize the importance of practicing good food safety behaviors by not consuming substances used as part of the lab investigation.
2. Distribute lab materials. It is recommended that materials are organized into stations for easier distribution. Students should be arranged in small groups of 4-5. Each group should receive the lab supplies outlined in the Materials section as well as 1 copy of the Hidden Antioxidants lab sheet.
3. Launch the lab by asking students to make a prediction about the antioxidant potential of various acidic substances.
4. Students should choose one substance to test their prediction. After obtaining and recording observational data within each group, allow the groups to share their findings with the class. This step is necessary for each student to complete Table A.
   • Tip: Groups may be assigned their test substance if it is desired that all substances be tested.
5. Refer to the following information for a list of expected reactions:
   • Cream of Tartar: Iodine should change the color of the solution to an opaque golden yellow (pH ~ 4).
   • Vinegar: Iodine should change the color of the solution to a medium clear, dark, golden yellow (pH ~ 3).
   • Vitamin C: Iodine should disappear when added to the solution. The solution should remain clear. However, if iodine is continually added, eventually the iodine will completely oxidize the vitamin C present, potentially resulting in a color change (pH ~ 3, slightly lower than vinegar).
   • Lemon Juice: Iodine should disappear when added to the solution. The solution should remain its original color. However, if iodine is continually added, eventually the iodine will completely oxidize the vitamin C present, potentially resulting in a color change (pH ~ 2).
6. Show students the video lab demonstration, Part II: Hidden Antioxidants.
7. Iodine can be used to detect vitamin C. When iodine is added to a vitamin C containing solution, ascorbic acid (vitamin C) is oxidized to dehydroascorbic acid and iodine is reduced to iodide ions.
   • C6H8O6 (Ascorbic acid) + I2 (Iodine) → 2 I- (Iodide) + C6H6O6 (dehydroascorbic acid)
8. Vitamin C tablets and lemon juice are the two substances with antioxidant properties. Vitamin C is an antioxidant and is contained in both substances. Oranges are also high in Vitamin C. In Part I-B of this lab, students should have observed the vitamin C solution and lemon juice prevent oxidative browning in apples. Other substances (e.g. sugar) can prevent oxidation browning; however, they work through other mechanisms such as reducing exposure of the fruit’s flesh to oxygen or lowering the pH.
9. Allow students to work in small groups to complete the Hidden Antioxidants lab sheet.
10. Follow-up with a class discussion about antioxidants and their role in human health.

Investigating Your Health: Amazing Antioxidants

1. Give each student one copy of the Amazing Antioxidants handout.
2. Using the information provided in the Background Agricultural Connections section and/or information learned from researching antioxidants, students should examine the food labels for a single fruit packaged in three different ways (i.e. frozen, dried, and canned).
3. Students can find food labels in the grocery store, USDA’s FoodData Central, or use the labels provided.
4. If you choose to use the provided pineapple food labels, see the attached teacher key for answers to the Investigating Your Health lab questions. Answers to questions based on other food labels will vary.
5. This assignment can be completed in-class or as homework. If completed in-class, allow students to work in small groups on the Investigation worksheet to further explore the topic and respond to questions.
6. Follow-up with a class discussion about student findings related to the health benefits of fruits and student generated ideas for increasing fruit consumption.