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Cady Bright
4.12.18
Chemistry 11

## How Much Sugar is in my Drink?

Absorption Spectroscopy Lab

Pre-Lab Questions
1. Define the following terms in the context of this investigation
A standard solution is a substance with a precise known concentration that, in the context of this lab,
can be used to create a calibration curve based on relative light absorbance as measured by a
spectrophotometer, which is a device that measures light absorbance. A calibration curve is essentially a
scatter plot made up of points representing known concentrations, and a linear line of best fit that reflects
them. To calibrate a spectrophotometer, a blank, or a control substance containing no concentration of the
solute, must be inserted into the device.

2. Define solute, solvent, solution, and concentration in the context of this investigation. Give a clear contextual
example of each.
A solution is made up of two parts, the solute and the solvent. The solute, in this case Kool-Aid
powder, is what is being dissolved into the solvent. The solvent, in this case water, is therefore what the solute
is dissolving into. A solution can be described using concentration, or mass of the solute divided by volume of
the total solution. In this investigation, we used grams of Kool-Aid powder divided by mL of the total solution.

3. What are the independent variable and dependent variable in this investigation?
The independent variable in this investigation was the Kool-Aid concentration - we changed the
grams of powder while the mL stayed the same, resulting in a change of concentration. The dependent was
then the absorbance of light being transmitted through the solution, which we measured using the
spectrophotometer.

4. When plotting a calibration curve, which variable goes on which axis?
In most investigations, the independent variable falls on the x axis and the dependent variable goes
on the y axis. This investigation was no different. As can be seen in figures 1-3, the independent variable -
concentration - lies on the x axis while the dependent variable - absorbance, lies on the y axis.

5. Explain how a calibration curve can be used to determine the concentration of an unknown solution. Be specific.
A calibration curve is created by plotting known concentrations against their respective known
absorbance values and creating a line of best fit that reflects them. Unknowns can then be analysed using a
spectrophotometer to find the light absorbance value, and placed on the line of best fit using that value. The
value would be the y-coordinate, and one could either use the equation for the line of best fit or one of various
computer graphing programs to come up with the corresponding x-coordinate, or concentration value.

6. Explain the basic operation of the spectrophotometer.
To operate a spectrophotometer, a cuvette, or small, clear container that resembles a rectangular
prism, filled with a blank substance must first be put in to calibrate it. The spectrophotometer sends a specific
amount of light through it, measures how much of the light was retrieved on the other side of the cuvette, and
sends that information to the computer. In the LoggerPro computer program, the concentration can be set to
zero, so the first point on the graph is the 0 concentration x, and whatever tiny number the spectrophotometer
records y. Then, solutions with higher concentrations are placed in a cuvette and from there into the
spectrophotometer, where light is sent through it, and the light retrieved is placed on the graph as the y value
to the manually inserted concentration (x) value.

Results and Analysis
1. Display the plot created using LoggerPro or create an equivalent plot using Excel or Google Sheets. Make sure
the axes contain proper labels with units. Make sure that the line of best fit is plotted and that the equation for
the line is present. If using LoggerPro state the RMSE and correlation value. If using Excel or Google Sheets state
the r​2​ value.
Figure 1: Concentration (g/dL) vs Absorbance at 380.2 nm

y = 0.05196x + 0.009724 , correlation value = .9998, RMSE = 0.008621

## Figure 2: Unknown A  Figure 3: Unknown B

0.638  0.398

2. The concentration of the unknown falls within the range of the concentrations of the standard solutions.
Therefore, the concentration of the unknown can be determined by interpolating from the calibration curve. To
do this, you would solve for the x-value (concentration of sugar in solution in g/dL) in the equation of the line of
best fit using your measured y-value (absorption). Show all work.
y = 0.05196x + 0.009724

Unknown A:
0.638 = 0.05196x + 0.009724
0.638 − 0.009724 = 0.05196x
0.628276 = 0.05196x
0.628276/0.05196 = x
12.0915319 = x
12.09g/dL

Unknown B:
0.398 = 0.05196x + 0.009724
0.398 − 0.009724 = 0.05196x
0.388276 = 0.05196x
0.388276/0.05196 = x
7.472594 = x
7.47g/dL

3. Compare your experimentally determined concentration value to the accepted value of the unknown solution
concentration that will be provided by your instructor using a percent error calculation. Show all work.
Unknown A:
|(12.09 − 12)/12| * 100 = .75%

Unknown B:
|(7.47 − 8)/8| * 100 = 6.625%

4. Describe the precision and accuracy of this method for determining the concentration of the Kool-Aid. Your
answer should consider the equation line of best fit, its measures of consistency (correlation value, r​2​ value and
RMSE), the methods employed in making standard solutions, how accurately your measured values of
unknowns match the expected values and the precision of the instruments used to measure absorption.
Overall, this method has shown to be quite accurate and precise in determining concentration using
absorbance measured by a spectrophotometer. Macroscopically, our procedure, while by human nature not
perfect, had little to no noticeable error. Our greatest difficulties were mixing the solvent and solute together
completely, and making the meniscus be where we wanted it to. The line of best fit appears to reflect the data
set well, when compared to others I have seen in this investigation.
Numerically, this method also appears very precise. Our correlation value was .9998, as measured on a
scale from -1 to 1 with 1 being a perfect positive association. Our RMSE value was 0.008621, which is also quite a
small margin of error. Perhaps the most accessible measure of our accuracy came when we tested the
unknowns. By using our previously created calibration curve to come up with the values for the unknowns and
then comparing them to the actual values utilising the experimental error equation, it can be again seen that
this method was quite precise. For Unknown A, our experimental error was .75%, while for Unknown B, it was
6.625%. Both of which are very low, however it would have been interesting to look further into why the
experimental error for Unknown B was higher.
A lot of our success lay in the equipment we were using, as the scales and spectrophotometers we
used are also used in college-level and higher settings. In fact, we had the most trouble being accurate with
getting exactly 1oo mL of solution in the flasks we were using. In the end, our struggles didn’t seem to affect
the investigation that much, and our methods proved reliable.