Chemisty 101 Last Name Last Name (Partner) ____________ ____________ First Name First Name (Partner) Lab

Time ____________ ____________ ____________ ___________ (Locker No.) ___________ (Locker No.)

Lab Day ____________ (M,T,W,R,F)

VITAMIN C IN A VITAMIN TABLET & IN FRUIT JUICE

Revised 3/24/2006

INTRODUCTION Vitamin C is one of the essential vitamins required for good health. Most people obtain the necessary amount of vitamin C in their diet (primarily in fresh fruits, fruit juices, and vegetables), but vitamin C tablets are also widely used as nutritional supplements. In this experiment, you will analyze a vitamin C tablet to determine the amount of vitamin C it contains. You will also be given data in order to calculate how much vitamin C is contained in a sample of fruit juice. Vitamin C, or ascorbic acid (C6H806), is chemically similar to the simple sugar, glucose (C6H1206), which is plentiful in our bodies. Most animals possess an enzyme needed for making ascorbic acid from glucose, but humans and a few other species lack that enzyme and therefore must secure ascorbic acid directly from foods they consume. The best-known function of ascorbic acid is the prevention of scurvy; a minute amount (10 mg per day) is adequate for this purpose. However; it also plays several other important roles in human health. Most of its functions are related to the fact that it has a strong tendency to transfer electrons to other chemical substances (chemists say that it is a "reducing agent" or "antioxidant"). It acts to prevent other, potentially harmful, reactions that involve the transfer of electrons.
B B B B B B B B B B B B

To maintain good health, the current recommended daily allowance (U.S. RDA) for vitamin C is 60 mg. Foods like fruits and vegetables with a high water content often contain large amounts of vitamin C. Vitamin C is very soluble in water, which means that, if high doses of vitamin C are ingested, much of it is rapidly excreted in urine. The experimental method that you will be using to determine the amount of Vitamin C in a vitamin tablet is called a titration. A titration is a laboratory procedure for doing a quantitative analysis (a determination of the exact amount of a substance in a given sample). In a titration, two reagents are mixed, one with a known concentration (the standard reagent) and one with an unknown concentration (the unknown). At the end point (or equivalence point) of the titration, where the chemical reaction has been completed, it is possible to calculate the exact amount of substance present in the sample of unknown concentration. In a titration, there must be some way to indicate when the two reagents have been mixed in the correct amounts to allow a complete chemical reaction between them. For this purpose, an indicator is added. This is a chemical substance that has one color before the chemical reaction 1

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has been completed and a different color when an excess of the standard reagent has been added. At the end point, we observe a transition from the first color to the second color. In a titration, the standard reagent is added carefully until one reaches the exact point when the color transition of the indicator is observed. At the end point, the number of molecules of the standard reagent is exactly equal to the number of molecules in the solution that we are analyzing.
TPF

The titration analysis for ascorbic acid (vitamin C) is based on its tendency to lose electrons. A suitable electron acceptor substance, in this case iodine (I2), is used as the reagent to "titrate" the ascorbic acid. Although it is not necessary to understand the reaction in order to do the analysis, the chemical equation for the reaction is the following:
B B

C6H806 + I2 C6H606 + 2 I- + 2 H+
B B B B B B B B B B B B B B P P P P

Since it is difficult to prepare iodine solutions with an accurately known concentration, the iodine concentration is determined first by titrating a reference sample of ascorbic acid. The "end point" of the titration is signaled by the appearance of the color of unreacted iodine, which occurs just past the endpoint. The color is made more intense by adding starch solution, which forms a deep blue color with iodine. OVERVIEW OF THE EXPERIMENT (SEE FLOW CHART BELOW FOR STEPS 2-4) 1. Assemble all of the necessary materials (see p. 3). 2. Titrate the ascorbic acid reference solution with iodine solution. 3. Prepare a solution of a vitamin C tablet in a volumetric flask. 4. Titrate portions of the vitamin C tablet solution with iodine solution. 5. Calculate the amount of ascorbic acid in the vitamin C tablet. 6. Calculate the amount of ascorbic acid in fruit juice with data given in lab. Titrate with ascorbic acid solution (concentration known): Step #2

Iodine solution (concentration unknown)

Iodine solution (concentration known) Titrate with (Step #4) Vitamin C solution (concentration known)

Vitamin C solution (concentration unknown) Step #3

Important note: In order to facilitate the calculation of mg vitamin C in the tablet and fruit juice, we will be using the unit mg/mL instead of mol/L (M) to describe concentrations of the solutions in this experiment. 2

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MATERIALS NEEDED Chemicals vitamin C tablet, 100 mg ascorbic acid reference solution iodine solution starch solution deionized (DI) water Equipment 4 15x125 test tubes 2 graduated-stem plastic pipettes 100-mL volumetric flask with cap 50-mL and 100-mL beakers

During the titrations, place a piece of white paper under the test tubes so that it will be easier to see the color changes as you do the experiment.

EXPERIMENTAL PROCEDURE I. Preparing the Vitamin C Tablet Sample 1. Weigh a vitamin C tablet on an analytical balance and record its mass to the nearest 0.0001 g. 2. Place the tablet inside a folded piece of smooth notebook or printer paper. Use the curved edge of the padlock from your lab drawer to crush it to a fine powder. 3. Carefully pour the entire powdered tablet from the paper to a clean 100-mL volumetric flask. You can use your funnel to assist in transferring the powder. Fill the flask about half-full with DI water. Swirl the flask well to dissolve the vitamin C tablet. Label the flask and set it aside to continue dissolving while you go on to the next procedure. II. Titration of the Ascorbic Acid Reference Solution 1. Using a clean, dry 50-mL beaker, obtain 10 mL of the ascorbic acid reference solution. Record its exact concentration (given in mg/mL) from the label. 2. Prepare four 15 x 125 test tubes, by labeling them #1 through #4. 3. Fill a graduated-stem plastic pipette with exactly 1 mL of the reference ascorbic acid solution. (It may take a little practice to fill the pipette exactly to the l-mL mark.) Then slowly dispense the l-mL sample into test tube #1. 4. In the same fashion, add exactly 1 mL of ascorbic acid reference solution to each of the remaining test tubes. 5. Add 3 drops of starch solution to each of the four test tubes. 6. Obtain 10 mL of iodine solution in a clean, dry 100-mL beaker. 7. Fill the other pipette with iodine solution. 8. Proceed to titrate the first ascorbic acid reference sample with the iodine solution, counting drops and stirring (use your spatula) as you go. You will observe a momentary gray/blue color that disappears upon stirring but lingers longer as more iodine is added. The end point occurs when one drop gives a pale grey/blue color that does not disappear upon stirring. Read the tips below before titrating. Record your data in the table on page 5. 3

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Designate a large beaker for waste to collect partial drops and to dump test tubes after completing sets of titrations. Remember that the most consistent drop sizes are obtained by holding the pipette vertically. It is best if you insert the tip of the pipette deep into the test tube in order to avoid getting the iodine solution on the side walls of the test tube. If you miss the endpoint or lose count of drops, simply go on to another sample. It is easy and fast to titrate additional portions of the solution as needed.

9. In the same fashion, titrate at least 3 more ascorbic acid reference samples. Record the results in the table on page 6. 10. Dump your titration waste and the remaining reference solution into your waste beaker. Carefully clean the four test tubes and the 50-mL beaker so that they can be reused. It is not necessary to dry them, but be sure to allow the excess water to drain out of them.

III. Titration of the Vitamin Tablet Solution 1. At this point, the vitamin C tablet in the volumetric flask should be mostly dissolved except for the insoluble "filler" material in the tablet. 2. Add DI water to fill the flask up to the bottom of the neck, then slowly add more DI water to bring the water level up the line etched on the neck of the flask. The last milliliter or so should be added one drop at a time. The bottom of the curved meniscus should just touch the line as shown in the following figure.
Water surface ("meniscus")

Etched line

Neck of volumetric flask

3. Put the cap on the flask securely and mix the contents of the flask thoroughly by repeatedly turning the flask upside down and swirling. (Hold the cap with a finger.) 4. Take the graduated-stem pipette that was used for the reference solution. Rinse it thoroughly with DI water (filling and emptying several times) and then rinse it with the solution from the volumetric flask, emptying the solution into your waste beaker, not back into the volumetric flask. 5. Use this pipette to add exactly 1 mL of the vitamin tablet solution to each of the four clean test tubes, as in Part II. 4

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6. Add 3 drops of starch solution to each of these samples. 7. Using the same iodine solution and plastic pipette that you used in Part II, titrate the first sample to the pale grey/blue endpoint. Note: Sometimes this endpoint may appear peach in color. Record the result in the table on page 7. 8. Repeat the titration of the vitamin C tablet solution at least 3 more times and record the results in the table on page 7. 9. Dump your titration waste and any excess solutions into your waste beaker, and dump them into the appropriate waste container in the waste hood. 10. Be sure to clean and return the 100-mL volumetric flask and the mortar and pestle along with the plastic pipettes.

VI. Data & Calculations show all work for each calculation!! Before analyzing your data, look carefully at the sets of titration data to see whether the results show consistency or whether any individual result should be eliminated because it appears to be very different from the other data in the set. If so, make a note beside that value and omit it from your calculations.

Titrations of the Reference Solution (Part II) Concentration of reference ascorbic acid ______________mg/mL (from label of bottle) Trial mL of reference solution Drops of iodine Note: Extra columns are for optional additional titrations. Average drops of iodine solution = _______________per 1 mL of reference solution Calculation: #1 1.00 #2 1.00 #3 1.00 #4 1.00 #5

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Calibration factor (see below) = _______________mg of ascorbic acid per drop of iodine Calculation: divide the ascorbic acid reference solution concentration (milligrams of ascorbic acid per milliliter from label) by the average number of drops of iodine calculated above (Equation 1). This calculation gives the mg of ascorbic acid corresponding to 1 drop of iodine solution. This is called the calibration factor for this analysis. Enter this number above. It is unique for the particular iodine solution and your equipment.

mg of ascorbic acid 1 mL of reference solution mg of ascorbic acid = 1 mL of reference solution drops of iodine 1 drop of iodine

( Eq. 1)

Titration of Vitamin Tablet (Part III)

Mass of tablet:________________ Titration mL of sample Drops of iodine Average drops of iodine solution per 1 mL of sample: _______________ Calculation: #1 1.00 #2 #3 #4 #5

mg of ascorbic acid per 1 mL sample (see below): _______________ Calculation: Multiply the average number of drops of iodine used to titrate the vitamin C solution by the calibration factor determined above to obtain the mg of ascorbic acid per mL (Equation 2). Enter the answer above. drops of iodine mg of ascorbic acid mg of ascorbic acid = 1 mL of sample 1 drop of iodine 1 mL of sample (Eq. 2)

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mg of ascorbic acid per tablet: _______________ Calculation: Use the milligrams ascorbic acid in 1 mL of the vitamin tablet solution and the total volume of the tablet solution to calculate the milligrams ascorbic acid in the whole tablet. Include units to help you keep track of your calculations.

Analysis of a Juice Sample

Three 1-mL portions of cranberry juice were titrated the same way you titrated the vitamin C tablet solution. The drops of iodine solution used are given in the table below. Using this data, complete the calculations below. Titration mL of sample Drops of iodine #1 1.00 7 #2 1.00 8 #3 1.00 7

Average drops of iodine solution per 1 mL of juice: ______________ Calculation:

mg of ascorbic acid per 1 mL: ______________ Calculation: Use Equation 2 above to calculate the milligrams of ascorbic acid per milliliter of juice. This time use the average number of drops of iodine used to titrate your juice sample.

mg of ascorbic acid per 8 ounces: ________________ Calculation: If a standard serving of juice is 8 fluid ounces and 1 fluid ounces is 30 mL, calculate the milligrams of ascorbic acid obtained from an 8-ounce serving of this juice.

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VIII. Questions (to be answered after completing the experiment)

1. Based on the experimental procedure used in this experiment, do you think vitamin C is a water-soluble or fat-soluble vitamin? Explain your answer and how it relates to the procedure.

2. What percent of the mass of the vitamin tablet was ascorbic acid? Show your calculation below.

3. Calculate the percentage difference between the milligrams ascorbic acid per tablet reported on the Vitamin C bottle label and the milligrams per tablet that you found experimentally (calculated at the top of page 7). % difference = mg ascorbic acid ( from VitC label ) mg ascorbic acid ( from analysis ) 100% mg ascorbic acid ( from VitC label )

4. The current (2002) U.S. RDA (recommended dietary allowance) for vitamin C is 75 mg/day for young adult women and 90 mg/day for young adult men. How much of the juice analyzed would you have to drink to meet the U.S. RDA for vitamin C? (Note: some nutritionists think the RDAs for vitamin C are too low and that 200 mg/day is more nearly optimal.)