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13D: Determining the Amount of Vitamin C

How can we determine the amount of ascorbic acid in vitamin C tablets and in different foods?
Ascorbic acid, or vitamin C, is very important for our bodies. Vitamin C is required by our bodies to produce collagen in our connective tissues. Vitamin C thus helps us to maintain healthy teeth, blood vessels, and bones. Vitamin C has also been found to function as an antioxidant, and it aids in the prevention of cancer. Antioxidants are molecules that can protect our cells from being damaged. Antioxidants such as vitamin C are able to absorb harmful reactive molecules before they can damage our tissues. In this investigation, a titration will be used to determine the amount of vitamin C that is contained in vitamin C tablets and in different foods and juices. The recommended daily amount of vitamin C is 60 mg.

Materials
250 mL beaker or Erlenmeyer flask Vitamin C solution (2 mg/mL), obtained by grinding one 500 mg tablet and dissolving in 250 mL of distilled water Dropper bottle 10 mL graduated cylinder 10 g of starch Distilled water bottle 20 mL of 0.010 M KIO3 solution 1.0 g KI 5.0 mL 1.0 M HCl Orange juice and grapefruit juice Citrus fruits and juicer Cheesecloth Medium funnel

We will determine the amount of vitamin C using a simple titration method. The chemical name for vitamin C is ascorbic acid. This procedure relies on a series of chemical reactions that cause an observable color change to occur. To help you better understand what you are seeing when the color change occurs, it is helpful to review some relevant chemical reactions. In equation two, Vitamin C reacts with iodine (I2) and becomes oxidized. This is the most important chemical reaction taking place. However, to produce the iodine (I2) we must first carry out another reaction. Adding solid potassium iodide (KI) and hydrochloric acid (HCl) and the titrant, potassium iodate (KIO3), creates iodine and water according to the following reaction:

1)

IO3 + 5I + 6H+ 3I2 + 3H2O

As soon as the iodine (I2) is created it reacts with the vitamin C, causing the vitamin C to become oxidized. 2)

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After all of the vitamin C has reacted, the I2 concentration builds up and then begins to react with I in solution to form the triiodide ion (I3). The triiodide ion complexes with the starch to give a deep bluishblack color that we can see: I2 + I I3 and I3 + starch iodidestarch complex deep blue color When this color appears, we know all of the vitamin C has reacted. It is the formation of the triiodide ion that signals the endpoint of the titration. Once we know how much titrant (KIO3) is required to reach the endpoint, we can use the volume of KIO3 to determine the moles of KIO3. Using stoichiometry we can use the moles of KIO3 to find the moles of iodine (I2) and therefore the moles of vitamin C present in our experiment.

Part 1: Calibrate your dropper bottle or pipette

1. We need to accurately determine the number of drops in 1.0 mL. To accomplish this, use either a dropper bottle or a plastic beral pipet. [In Part 2, be sure you add the titrant (KIO3) using the same dropper and method]. 2. To calibrate your dropper, slowly add drops of water to a small 10.0 mL graduated cylinder. Stop adding drops when the meniscus is on the 1.0 mL mark. Record the number of drops you added. 3. Next count how many drops it takes to get to the 2.0 mL mark. Record your measurement. If the number of drops is in close agreement, take the average. If your number of drops does not agree, repeat Steps 2 and 3.

Part 2: Verifying the amount of vitamin C in a commercial tablet

1. Measure 25.0 mL of the vitamin C stock solution, and add it to a 250 mL Erlenmeyer flask. Your instructor will have prepared this ahead of time. Record the concentration of the stock solution in your lab notebook. 2. Prepare a 1% starch solution. 3. Measure 1.0 g of KI solid and add it to the Erlenmeyer flask. Swirl to dissolve. 4. Add 5.0 mL of 1.0 M HCl to the Erlenmeyer flask. 5. Add 23 mL of the fresh 1.0% starch solution indicator to the Erlenmeyer flask. Swirl to mix. 6. Obtain a dropper bottle of 0.010 M KIO3 titrant solution. This dropper bottle should be the same type as the one you used in Part 1. If your instructor has the titrant in a stock bottle, measure about 25.0 mL of the 0.010 M KIO3 titrant solution in a clean, dry beaker and add it to a dropper bottle yourself. 7. Now you are ready to titrate. Add 0.010 M KIO3 solution drop by drop to the Erlenmeyer flask. Swirl after every few drops. Initially you will see the deep blue color appear in the area that the drop is added. Once you begin to see a deep blue color that persists throughout the solution, add the titrant more slowly and swirl after every drop. Continue adding titrant until you obtain a

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Investigation 13D: Determining the AmountBack of Vitamin C of Contents Print Start Menu Table permanent blue color that lasts about 60 s. Record the total number of drops you added. The solution may lighten after the 60 s, but it should remain fairly deep blue. Determining the right color takes practice and is somewhat objective. Do your best, and then repeat the experiment. After two or three trials, you will feel more confident at determining your endpoint.

Part 3: How much vitamin C is there in different fruits and juices?

Here we will determine the vitamin C content of a food of your choice. You may use citrus fruits, fresh squeezed or prepared, apple juice, or any drink containing ascorbic acid. Real lemon juice also makes a nice sample to test, and the amount of vitamin C is on the label. 1. To begin you may use fresh or prepared juice. You will need about 25 mL of juice to have enough vitamin C for a good measurement. If you use freshly squeezed juice, it is helpful to strain out the pulp with cheesecloth. An easy way to strain out the pulp is to use a funnel lined with two or three layers of cheesecloth. Pour the sample of juice through the cheesecloth and collect the juice. Record the exact volume of juice you will titrate. 2. Add your juice to a 250 mL Erlenmeyer flask. 3. Titrate your sample using the same steps you used to titrate the vitamin C tablet. Be sure to add 1.0 g of solid KI, 5.0 mL of 1.0 M HCl and 23 mL of starch solution. Note: You may not see the deep blue color because of the color of your fruit juice. For instance, it may appear brownish instead of blue. This is fine; just be sure you look for the same color change in each trial. That is the key. 4. Repeat your experiment two or three times to obtain an accurate sense of the reproducibility of your results. Record your results and find the average amount of vitamin C in your sample.

Part 4: Thinking about it

1. Make a data table that records the number of drops of 0.010 M KIO3 needed to titrate 25 mL of the vitamin C stock solution. 2. Convert your drops to milliliters using the conversion you determined in Part 1. Record this in your data table. Here is an example: 3. Does it matter how much distilled water was added to your sample? Explain. 4. Which contained more vitamin C: the tablet or the food? How do you know? 5. Look carefully at the first equation on page 113. Copy this equation into your lab notebook. Which substance was the reducing agent? Explain. Which substance was the oxidizing agent? Explain.
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Part 5: What did you obtain for results?

1. Use the volume of your KIO3 titrant added in liters and the known molarity (0.010 M) to calculate the moles of KIO3 required to titrate the vitamin C sample, as shown in this example: 2. Using the second equation on page 113 and stoichiometry, convert the moles of KIO3 to moles of I2 produced. See the example calculation shown in Step 3. 3. Calculate your moles of vitamin C, from the amount of I2 produced in Step 2. Use the first equation on page 113, stoichiometry, and the following example to help you with this calculation:

4. Calculate the grams of vitamin C from the moles of vitamin C you determined. 5. Convert your grams to milligrams by dividing by 1,000. This number tells you how many milligrams of vitamin C were in your 25.0 mL sample of the vitamin C stock solution, or your food. 6. Which food contained the highest amount of vitamin C? Explain. 7. The stock solution contained a 500 mg tablet of vitamin C and 250 mL of distilled water. Combine your results with those of your classmates and see how close your results came to this value. For example, if each group titrated 25.0 mL of stock solution, then the total milligrams in 10 trials would give you a total volume of 250 mL of solution, and this should give you a value close to 500 mg of vitamin C. To estimate, you could also take your trial average and multiply the number of milligrams by 10 to see if you are close to 500 mg. 8. Based on your result, does a 500 mg tablet really contain 500 mg of vitamin C? 9. If the recommended daily allowance of vitamin C is 60 mg, how much of your food would you need to consume to get your daily allowance? Show your calculation.

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