How Blue is Your Sports Drink?
05/09/17, 2*22 PM
How Blue is Your Sports Drink?
Experimental Procedure
The following procedure can be broken into three parts:
1. Build and test the spectrophotometer that converts the
concentration of dye in a solution into electrical resistance, which you
can read off a multimeter;
2. Make a set of standard solutions, so that you know how to convert
between the data you have (resistance) and the data you want
(concentration); and
3. Determine the amount of dye in your sports drink samples with
unknown concentrations of Blue 1.
Part 1: Building and Testing the Spectrophotometer
In this section, you will assemble a circuit on a breadboard. If you have
never used a breadboard before, you should refer to the the Science
Buddies resource How to Use a Breadboard before you proceed. You can
follow a step-by-step slideshow that will show you how to put
components in the breadboard one at a time. Alternatively, Table 1 lists
each component and its location on the breadboard. Important: Read
these notes before you proceed:
Resistors are marked with colored bands. These colors do matter.
Make sure you pick the right resistors for each step according to the
markings.
It matters in which direction some of the components are facing.
Make sure you read the slideshow captions for any special notes
about inserting each part.
This section only shows you how to assemble the circuit. For a
detailed explanation of how the circuit works, see the Help section.
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How Blue is Your Sports Drink? 05/09/17, 2*22 PM
Click through the slideshow to see the colorimeter procedure.
Table 1. Components for the spectrophotometer circuit (Image credits:
Jameco and Fritzing).
After you have finished building your circuit, testing the
spectrophotometer is necessary to ensure that all the electronic
components are connected correctly and your device works as expected.
1. Place one empty cuvette, upside-down, over the LED; and another
empty, upside-down cuvette over the photoresistor. If the cuvettes
are not clear on all sides, but have two grooved or frosted sides,
make sure that you put the clear side facing toward the LED as well
as the photoresistor. Bend the LED and photoresistor as needed to fit
underneath the cuvettes.
2. Place two empty cuvettes between the LED and the photoresistor.
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Again, make sure that you always face the clear sides of the cuvette
toward the LED and the photoresistor. The four cuvettes should
touch each other and form a straight line. You can use clear tape to
hold the cuvettes over the LED and the photoresistor in place. But do
not block the light path!
3. The light from the LED should now shine directly onto the
photoresistor, as shown in Figure 4. Bend the wires on the LED and
photoresistor for adjustment, if needed.
Figure 4. Make sure that the LED and the photoresistor are properly
aligned. Note that in this picture, the cuvettes are not yet placed on top of
the photoresistor and LED.
4. Set up the multimeter to measure the resistance of the photoresistor.
a. Plug the black multimeter probe into the port labeled COM.
b. Plug the red multimeter probe into the port labeled VMA.
c. Turn the dial setting to 200 ohms (the green "200" at the bottom
of the dial near the symbol).
d. Use alligator clips to attach the red and black multimeter probes
to the red and black jumper wires connected to the
photoresistor coming from A28 and J28.
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5. Turn on the LED by connecting the jumper wire from B15 to the
power (+) bus.
6. Cover the circuit (but not the multimeter) with the cardboard box to
block ambient light.
7. Read the resistance across the photoresistor and record it in your lab
notebook.
a. Note the units of the resistance. A "k" indicates kilo-ohms (k)
and an "M" indicates mega-ohms (M).
b. If your multimeter screen displays a "1 .", that means the
resistance is too high for the dial setting. Turn the dial up to the
next highest range (for example, from 200 to 2000) and check
again.
c. If this is your first time using a multimeter, refer to the Science
Buddies resource How to Use a Multimeter, specifically the
section How do I measure resistance?, to learn more.
8. Remove the box and turn off the LED by removing the jumper wire
from the power (+) bus.
9. Cover the circuit with the box again. In the dark, the resistance
should be in the mega-ohm range. Remember that you may need to
adjust the dial setting to get a measurement. Record the resistance in
your lab notebook. Note: Stray light will cause problems with the
data. Perform the readings in a dimly lit room if stray light is a
problem and/or use a black permanent marker to shield the
photoresistor from light from the sides and back of the cuvette.
10. Remove the box and turn off the multimeter to conserve battery
power.
Part 2: Calibrating the Spectrophotometer
Now that you know that your spectrophotometer is working, the next step
is to make the standard solutions to calibrate it. You will make a series of
dilutions of blue dye, as shown in Figure 5, with known concentrations,
and measure them with your spectrophotometer to create a calibration
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curve. Each dilution is made by consecutively diluting your solution by
half. It is essential to use dye-free utensils and cups to get an accurate set
of standards.
Figure 5. Standard solutions of Blue 1 for creating the calibration curve.
1. Set out eight clean cups and label them 18.
2. The solutions will be diluted as follows:
1 (most concentrated)
1/2
1/4
1/8
1/16
1/32
1/64
Water only
3. Pour 8 oz. of water into the first cup (#1) and 4 oz. of water into the
remaining cups (28).
4. Mix 1/8 teaspoon (tsp.) of blue dye with the 8 oz. of water in cup #1.
Note: The concentration of blue dye in the commercial package is
approximately 0.026 M (mol/L). After dilution (1/8 tsp in 1 cup =
1j384), the concentration is 68 M (mol/L).
5. Stir the contents of cup #1 with a clean spoon.
6. Using the measuring cup, pour 4 oz. from cup #1 into cup #2 and mix
with a clean spoon.
7. Thoroughly rinse the measuring cup and spoon and mix 4 oz. from
cup #2 with the water in cup #3.
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8. Repeat the two-fold dilutions for cups 47. Cup #8 will be your
"blank," and should not contain any dye.
9. Transfer the blank and the standard solutions into eight clean and
labeled cuvettes. Use the eyedropper, a transfer pipette, or pour
carefully. Note: The cuvettes hold approximately 3 mL of solution.
10. Prepare your orange absorption filter by adding 120 mL or 1/2 cup of
water into a clean cup. Add two drops of red and two drops of yellow
liquid food coloring and mix the solution well with a clean spoon.
11. Transfer the orange solution into a clean cuvette and place the
cuvette next to the LED so that the clear sides face the LED and the
photoresistor.
12. Attach the red and black multimeter probes to the red and black
wires in contact with the photoresistor (coming from A28 and J28)
using the cables with the alligator clips, if they are not yet connected.
13. Set the multimeter to read resistance again. Remember that you
might have to adjust the range as you take different readings.
14. First, place your blank sample without dye in between the orange
cuvette and the photoresistor, as shown in Figure 6. Again, the clear
sides of the cuvettes should face toward the LED and the
photoresistor.
Figure 6. Spectrophotometer setup for measuring your blank (left),
standard, and samples (right). Note that in these pictures, the LED is not
yet switched on. For your measurements, you also have to cover the
spectrophotometer with a cardboard box to block out surrounding light.
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15. Plug in the wire to turn on the LED and cover the breadboard with a
small cardboard box. Read the resistance on the multimeter and
record the data in your lab notebook.
16. Remove the blank cuvette and replace it with the cuvette containing
the next standard solution, starting with the lowest concentration.
Cover the breadboard again with the cardboard box and write down
the resistance for this solution. Continue the measurements for each
of your seven standards.
17. Repeat steps 1416 with the entire set of standards, including the
blank, two more times.
18. Make a data table in your lab notebook, showing the dilutions and the
concentrations of blue dye in all your standards (#1 = 68 M, #2 = 34
M, etcetera) together with all three recorded resistance
measurements for each solution. The resistance should be higher as
the solutions get darker.
Part 3: Measuring Your Sports Drink Samples
You are now ready to take readings from your spectrophotometer with
real sports drink samples.
1. Start with a visual evaluation of each of your blue beverages. Which
one do you think contains the most amount of blue dye? Write down
your assumptions in your lab notebook.
2. Label as many clean cuvettes as you have sports drinks that you
would like to test. Make sure the label is explicit for each drink.
3. Using the clean eyedropper or a transfer pipette, fill each clean
cuvette with one of the blue-colored beverages, such as in Figure 7.
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Figure 7. Several sports drink samples with unknown concentrations of
Blue 1 prepared for measurement on the spectrophotometer.
4. Check that your spectrophotometer setup is still in measuring mode
with the leads attached to the multimeter, and both the LED and
multimeter switched on. Place the cuvette with the orange solution
next to the LED if it is not already there.
5. Place one of your sports drink samples on the device in between the
orange filter solution and the photoresistor. The clear sides of the
cuvette need to face the LED and the photoresistor.
6. Cover the spectrophotometer with the cardboard box and record the
resistance on the multimeter in your lab notebook. Note: If the
resistance of your solution exceeds the maximum resistance of your
calibration curve, dilute your sample and measure again. You can do
a 1j2 dilution in a fresh cuvette (1.5 mL water + 1.5 mL sample
solution) or a 1j6 dilution (2.5 mL water + 0.5 mL sample solution).
7. Measure your sample two more times.
8. Continue measuring all your sports drink samples on the
spectrophotometer and record the resistance for each in your lab
notebook. Be sure to measure each sample a total of three times.
Analyzing Your Results
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1. Open a spreadsheet and enter the resistance data for your calibration
curve. Calculate the average for your three resistance readings for
each standard. Subtract the resistance that you measured for the
blank from all of the readings you made for samples with dye. This
step subtracts the light loss due to the plastic, the water, and other
factors.
2. Graph the average resistance of your three readings on the y-axis
versus the concentration of the standard solutions in M on the x-
axis. Note: If you are using Microsoft Excel, use the "Scatterplot"
chart. Excel also has tools for adding trend lines.
3. More-advanced students can add a trend line to the data and display
its equation and its correlation factor R2.
4. Graph the average resistance of each of your blue beverages on the
chart.
5. Determine the concentration of blue dye in your sports drink
samples, based on where they are on the graph or use your
calibration curve to calculate the concentration of blue dye in your
blue beverages. Remember to account for your dilutions if a sample
had to be diluted.
6. Which of the beverages had the highest concentration of blue dye?
Do your results agree with your visual evaluation of the sports drinks?
Troubleshooting
For troubleshooting tips, please read our FAQ: How Blue is Your Sports
Drink?.
Communicating Your Results: Start Planning Your Display
Board
Create an award-winning display board with tips and design ideas from
the experts at ArtSkills.
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