Food Colorant Lab Report

Aaron Mata

Sutton Vargo

Lab Section 1215 - 015

10/29/2019
Introduction

Absorption Spectroscopy is used in order to determine the colorants in the name brand

beverage. Different atoms will absorb different wavelengths of light and that will allow for the

absorbance to be recorded. Because absorbance and concentration are directly proportional, by

comparing absorptions it will be possible to determine the concentrations of the dyes that are

present in the name brand beverage in order to create a brand-new generic beverage with the

same color as the name brand beverage.

Light absorption is created when electrons start from their lowest state and jump to a

higher energy level while absorbing energy from photons of light. This energy can be calculated

using the equation E=hc/λ where h is Planck’s constant (6.626E- 34Js), c is the speed of light

(3.00E8m/s), and λ is the wavelength of the photon absorbed (m). Each atom has different spaces

in between these energy levels therefore resulting in different wavelengths that will then result in

different absorbance values from each given atom.

The Beer-Lambert Law help understand the fact that the relationship between

concentration and absorption are directly proportional. Depending on the ions within a solution,

it will determine how much light is absorbed in the solution and will lead to the concentration of

those ions within the solution. The equation of Beer-Lambert Law is A= εbc, where A is the

absorbance, ε is the molar absorptivity, b is the path length, and c is the concentration. Because

the absorbing species and the cuvettes will be the same, both constants b and ε will form one
constant leaving concentration left to solve, given that the absorption of the species is

determined.

Experimental

Reagents used include the following:

Deinonized Water

Solution of FD&C Blue #1 (6.75 x 10-6 M)

Solution of FD&C Red #40 (4.00 x 10-5 M)

Solution of FD&C Yellow #5 (4.00 x 10-5 M)

Name Brand Beverage

Calculate 4 solution concentrations, M, of every dye as well as the beverage use the

formula M1V1 = M2V2. Begin by coming up with 4 separate concentrations and using the 10mL

Volumetric flask to create each solution with the proper volume, of the given dyes as well as the

water to dilute it, calculated by the formula mentioned above.

Once each solution has been created proceed to record an absorption spectrum from the

solutions. Set up the MeasureNet station to record the absorption of each solution and transfer

the solutions created into a clean and dry cuvette at least ¾ full. Proceed to the spectrometer and

press the given buttons associated with the proper station number. Furthermore, zero out the

spectrometer and place the light block cuvette given, after cleaning it with a Kimwipe, into the
sample holder and then press “zero.” Then replace the light block cuvette with one cuvette of

simple distilled deionized water to serve as the reference cuvette.

After creating the reference cuvette, run the sample created from the solutions mentioned

above. Repeat this step for the rest of the other solutions/samples as well as the solution with the

name brand beverage. Save every data collected to the proper station number assigned. Proceed

to clear up the station and empty the sample cuvettes into the proper liquid waste container.

Email the data files to each lab partner in order to create absorption plots.

Using the data collected, 8 total absorption lined scatter plots will be created while

properly labeling the titles, axis titles and units. Then the lambda max will be determined along

with the associated absorbance value from each solution in order to create a beer-lambert plot.

Furthermore, a linear regression analysis on the plot will be created including the R^2 value and

the linear equation on the plot. The absorbance value given from the unknown solution and the

linear equation on the Beer-Lambert plot will allow for the concentration of the solution to be

determined.
 Results

Absorbance of Concentration of Name Brand Unknown

Yellow Yellow (M) Beverage Concentration of
Absorbance of Yellow (M) in name
Yellow brand beverage
0.461 3.00E-05 0.509 9.00E-05(.509) – 2.58E-05
2.00E-05 =
0.448 2.00E-05
0.392 1.00E-05
0.242 5.00E-06
Lambda Max = 424.29 nm

Absorbance of Concentration of Name Brand Unknown

Blue Blue (M) Beverage Concentration of
Absorbance of Blue (M) in name
Blue brand beverage
0.691 5.00E-06 0.209 4.00E-06(.209) + 2.84E-06
2.00E-06 =
0.300 4.00E-06
0.138 3.00E-06
0.204 2.00E-06
Lambda Max = 628.86 nm

Blue Sample 0.716

R^2=

Yellow Sample 0.726

R^2=
 BLUE SAMPLE BEER-LAMBERT PLOT (LAMBDA MAX
y = 4E-06x + 2E-06
OF 628.86 NM) R² = 0.7161

6.00E-06

5.00E-06

4.00E-06
ABSORBANCE

3.00E-06

2.00E-06

1.00E-06

0.00E+00
0.13 0.23 0.33 0.43 0.53 0.63
CONCENTRATION (M)

YELLOW SAMPLE BEER-LAMBERT PLOT (LAMBDA

y = 9E-05x - 2E-05
MAX OF 424.29 NM) R² = 0.7265
3.50E-05

3.00E-05

2.50E-05
ABSORBANCE

2.00E-05

1.50E-05

1.00E-05

5.00E-06

0.00E+00
0.24 0.29 0.34 0.39 0.44
CONCENTRATION (M)
 Absorbance Absorbance Absorbance

-0.2
-0.1
0
0.1
0.2
0.3
0.4
0.5
0.6
-0.1
0
0.1
0.2
0.3
0.4
0.5
0.6
-0.2
0
0.2
0.4
0.8
1

-0.2
0.6
180.28 180.28 180.28
207.6 207.6 207.6
236.24 236.24 236.24
264.67 264.67 264.67
292.89 292.89 292.89
320.89 320.89 320.89
348.68 348.68 348.68
376.24 376.24 376.24
403.58 403.58 403.58
430.68 430.68 430.68
457.56 457.56 457.56
484.19 484.19 484.19
510.58 510.58 510.58
536.72 536.72 536.72
562.61 562.61 562.61
588.25 588.25 588.25
613.63 613.63 613.63

Wavelength (nm)
Wavelength (nm)
Wavelength (nm)

638.75 638.75 638.75

663.6 663.6 663.6
688.17 688.17 688.17
712.48 712.48 712.48

Absorbance Spectrum of Blue #300

Absorbance Spectrum of Blue #200
Absorbance Spectrum of Blue #100

736.5 736.5 736.5

760.24 760.24 760.24
783.7 783.7 783.7
806.86 806.86 806.86
829.73 829.73 829.73
852.3 852.3 852.3
 Absorbance Absorbance Absorbance

-0.2
-0.1
0
0.1
0.2
0.3
0.4
0.5
0.6
-0.1
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8

-0.2
-0.1
0

-0.2
180.28 180.28 180.28
206.09 206.09 206.09
233.23 233.23 233.23
260.19 260.19 260.19
286.96 286.96 286.96
313.54 313.54 313.54
339.93 339.93 339.93
366.11 366.11 366.11
392.1 392.1 392.1
417.87 417.87 417.87
443.44 443.44 443.44
468.8 468.8 468.8
493.94 493.94 493.94
518.86 518.86 518.86
543.56 543.56 543.56
568.03 568.03 568.03
592.28 592.28 592.28
Wavelength (nm)

Wavelength (nm)

Wavelength (nm)
616.29 616.29 616.29
640.06 640.06 640.06
663.6 663.6 663.6
686.89 686.89 686.89
Absorbance Spectrum of Blue #400

709.93 709.93 709.93

Absorbance Spectrum of Yellow #101

Absorbance Spectrum of Yellow #102

732.73 732.73 732.73
755.27 755.27 755.27
777.55 777.55 777.55
799.58 799.58 799.58
821.34 821.34 821.34
842.83 842.83 842.83
864.06 864.06 864.06
 Absorbance Absorbance

-0.2
0
0.1
0.2
0.3
0.4
0.5
0.6
-0.2
-0.1
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7

-0.1
180.28 180.28
203.06 203.06
227.22 227.22
251.23 251.23
275.09 275.09
298.8 298.8
322.36 322.36
345.76 345.76
369.01 369.01
392.1 392.1
415.02 415.02
437.78 437.78
460.37 460.37
482.79 482.79
505.04 505.04
527.12 527.12
549.02 549.02
570.74 570.74
592.28 592.28

Wavelength (nm)
Wavelength (nm)

613.63 613.63
634.8 634.8
655.78 655.78
676.57 676.57
697.16 697.16
Absorbance Spectrum of Yellow #103

Absorbance Spectrum of Yellow #999

717.56 717.56
737.76 737.76
757.76 757.76
777.55 777.55
797.14 797.14
816.53 816.53
835.7 835.7
854.66 854.66
 Absorbance Spectrum of Name Band Beverage
0.6

0.5

0.4

0.3
Absorbance

0.2

0.1

618.94
180.28
200.03
221.19
242.24
263.18
284
304.7
325.29
345.76
366.11
386.34
406.44
426.42
446.27
465.99
485.58
505.04
524.37
543.56
562.61
581.53
600.31

637.43
655.78
673.98
692.03
709.93
727.68
745.28
762.73
780.01
797.14
814.12
830.93
847.57
864.06
-0.1

-0.2
Wavelength (nm)

Discussion

In the initial graph of the absorbance of the name band beverage with an unknown

concentration, it is clear that the composition is mostly that of blue dye, with some yellow, and

the tiniest bit of red when compared to the absorbance reference given by the instructor.

Therefore, it was decided to direct the experiment based around the colors blue and yellow. The

Beer-Lambert plots created from the multiple concentrations and absorbance recordings in each

individual dye allowed for a linear regression equation to be made. With this equation, the

absorbance reading from the initial graph of the unknown concentration, was used to

approximate the concentrations of each dye in the name brand beverage. The approximate

concentrations of each dye needed to create a brand-new generic beverage, were 2.58E-05M of

the yellow dye and 2.84E-06M of the blue dye.

 After doing the experiment, we are fairly confident with our results, however, there were

definitely errors that could have been made. With the tools that were given, it was impossible to

measure out each dye to the nearest thousandth. Given that, there was definitely room for some

error. We ended up doing our best to measure the dyes to the nearest tenth. I’m not sure that this

could have been avoided given our resources, but we think we measured with the best of our

abilities given the tools at hand.

In order to make a 10 mL solution with a color exactly matching that of the name brand

beverage a concentration of 2.58E-05M of the yellow dye and 2.84E-06M of the blue dye are

required. For a 10 mL solution, the equation M1V1=M2V2 is needed. For the yellow dye it

would be M2 = 2.58E-05M, V2=10mL and M1= 4.00 E1-05 M, therefore solving for V1 (in mL)

by multiplying M2 and V2 then dividing that number by M1, so there would need to be

approximately 6.45 mL of yellow dye in the solution. For the blue dye it would be M2 = 2.84E-

06M, V2 = 10mL and M1= 6.75E-06M, therefore solving for V1 (in mL) by multiplying M2

and V2 then dividing that number by M1, there would need to be approximately 4.21 mL of blue

dye for the solution to have the exact color that matches the name brand beverage.

Conclusion

The experiment yielded results that were helpful in answering the central question of what

concentrations of dyes present in the name brand beverage are needed in order to create a brand-

new generic beverage with the same color as the name brand beverage. The most important

results from the experiment were from the linear regression line of the Beer-Lambert plots
associated with both the blue and yellow concentrations. Those equations allowed us to use the

absorbance of each dye in the sample beverage recorded and determine the approximate

concentration of each dye in the name brand beverage.

Bibliography

David Thomas, D. S. (2019). Experiment C - Using Absorption Spectroscopy to Determine

Concentration. Salt Lake City.