Food Dye Analysis of Commercial Products

CHM 2045L Section 003

T.A. Timothy Vasquez
4/27/15

Introduction
Many people do not know that food dyes can potentially be damaging to the body. The
dyes used in this experiment Red #40, Yellow #5, and Yellow #6, are actually found to be tainted
with contaminants (Curran). These dyes, which are found in common food products, were tested
for. Red #40, the most-widely used dye, causes hypersensitivity (allergy-like) reactions in a small
number of consumers and might trigger hyperactivity in children (Jacobson and Kobylewski).
Yellow #5 may be contaminated with several cancer-causing chemicals. Also, it may cause
sometimes-severe hypersensitivity reactions in a small number of people and might trigger
hyperactivity and other behavioral effects in children (Jacobson and Kobylewski). Yellow #6
may also be contaminated with cancer-causing chemicals and occasionally causes severe
hypersensitivity reactions. Yellow #6 adds an unnecessary risk to the food supply (Jacobson and
Kobylewski).
In this experiment, spectrophotometry was utilized as an essential tool.
Spectrophotometry and UV-Visible radiation can be used in combination to determine
concentrations of food dyes. Food dyes absorb electromagnetic radiation in the visible spectrum
(Sandi-Urena et al). In this experiment, the interaction of UV-Visible radiation and organic
molecules in commercial food products were studied to determine concentration of certain food
dyes. From the given variety of dye colors, Red #40, Yellow #6, and Yellow #5 were compared
using an OceanOptics spectrophotometer. In combination with the use of spectrophotometry,
Beers Law was important in this experiment as well. Beers Law is described as A=bc where A
is absorbance, is the molar absorptivity that is, the path length of the cuvette in which the
sample is contained, and c is the concentration of the compound in solution (Blaunch). The main

goal of this lab was to find the unknown concentration of commercial food products containing
Red #40, Yellow #6, and Yellow #5.
Methods
Preparation of Stock Solution
First, the solution of Red #40 was prepared. 0.02 g of the solid dye was obtained and
mixed with 100 mL of Deionized water in a 150 mL beaker. The same amounts and procedures
were followed for Yellow #6 and Yellow #5. In order to determine which solution was to be used
as the stock, the absorbance was observed in a Spectrophotometer. If the absorbance was over 1,
the solution needed to be diluted until it reached an appropriate level of absorbance. After setting
up the spectrophotometer, separate cuvettes were prepared. One was filled with just deionized
water to be used as a blank when changing wavelengths. The other cuvettes were filled with each
according dye solution. For the dye solutions, they were diluted using serial dilution. 10 mL of
the original solution was taken and combined with 10 mL of deionized water. The solution was
continuously diluted until the absorbance was at or below 1, which would then be the stock
solution. The same procedure was performed on each dye until a stock solution was found.
Creating a Calibration Curve
After the stock solutions had been prepared, a calibration curve was created using 5
points. First, the maximum absorbance of each solution was needed to be found. Using the stock
solution of the specified dye, serial dilution was performed by taking 10 mL of the stock
(Solution 1) in combination with 10 mL of deionized water. This new solution (Solution 2)
would then be similarly tested in the spectrophotometer to find the maximum absorbance. 10 mL
of Solution 2 was then added with 10 mL of deionized water to make Solution 3. The procedures

were repeated using serial dilution until there were 5 solutions. Each solution was then tested for
maximum absorbance and recorded. When the Red #40 solution was put in the
spectrophotometer, the absorbance read high above 1. In order to bring it closer to 1, it was
diluted 3 times. Once the readings had fallen below one, that solution was used as the stock.
Similarly, with the Yellow #5 solution, it was diluted 3 times to find the stock solution. The
Yellow #6 solution was diluted once. The concentrations of the stock solutions were found by
halving the original concentration with each dilution. Then, the concentration of each solution
needed to be recorded. First the mass of the solid dye, 0.02 g, was divided by the molar mass
then divided by the volume used, 0.1 L, and the concentration was found. For each solution that
was diluted, the original concentration of the dye was divided in half. Afterwards, the calibration
curve was created by plotting the maximum absorbance against the concentration of each
solution.
Determining the Unknown
Different household food products containing at least the three given food dyes (Red #40,
Yellow #5, and Yellow #6) were brought for the experiment, which were M&Ms and Tic-Tacs.
The concentration of each product was needed to be found. By comparing the maximum
absorbance of the designated food product to the previously created calibration curves, the
unknown concentration could be found. First, the M&Ms, which contained both Red #40 and
Yellow #5, were tested by dissolving the color from the shell for 10 seconds while stirring in 100
mL of deionized water. This had to be done carefully so that no solid pieces broke off into the
solution. This method was repeated again for the second dye solution. Then, using the Tic-Tacs
for dye Yellow #6, one Tic-Tac was dissolved similarly to the M&Ms in 100 mL of deionized

water. Each food product solution was tested for maximum absorbance and then plotted along
the corresponding food dyes calibration curve to find the unknown concentration.
Results
Table 1: Calibration Curve of Red #40

Table 2: Calibration Curve of Yellow #6

Table 3: Calibration Curve of Yellow #5

Table 4: Comparison of class results

When creating the stock solutions, the original solution of 0.02 g of the solid dye in 100
mL of water was diluted. Before finding the stock solution, the original concentrations were
found using the molarity formula:
40=

0.02 g
1 mol

=0.0004
496.92 g /mol 0.1 L

5=

0.02 g
1 mol

=0.00037
535.3 g /mol 0.1 L

6=

0.02 g
1 mol

=0.00044
452.37 g /mol 0.1 L

The concentration of the stock solutions were determined after serial dilution had been

performed:

40 Stock Solution=

0.0004 0.0002 0.0001

=
=
=0.00005
2
2
2

5 Stock Solution=

0.00037 0.00019 0.00009

=
=
=0.000046
2
2
2

6 Stock Solution=

0.00044
=0.00022
2

In determining the unknown concentration (x) of the food products, the known
absorbance (y) was plugged into the linear equation of the calibration curve graph and solved
for:
Concentration of 40 M M ' s :0.2=16150 x0.0486, x=0.000015
Concentration of 5M M ' s : 0.27=13099 x+ 0.0784, x=0.000015

Concentration of 6TicTacs :0.18=4235.1 x +0.0183, x=0.000038

Discussion
In comparison of the three dyes used in this experiment to the class results (Table 4),
there were varying outcomes. The concentration of Red #40, while found to be 0.00005 M in this
experiment, Group 1 had a concentration of 0.00001496 M and Group 3 had a concentration of
0.00008508 M. In the experiment, Yellow #5 was found to be 0.000046 M. The other groups
found that at the stock solution, Group 3 had a concentration 0.00007488 M and Group 1 had
0.000161 M. For Yellow #6, the concentration was found to be 0.000022 M and Group 4 had
found 0.00010615 M and Groups 5&6 had 0.000631 M. The reason there could be such variation
in the concentrations across the different groups is that when diluting to find the stock solution,
different degrees of dilution were performed or general lab errors occurred.
The theoretical and experimental maximum wavelength (max) of the various dyes were
also compared (Table 4). Red #40 had a theoretical max ranging from 504-506 nm. The
experimental max was found around 505-509 nm, which is fairly close to the theoretical
wavelength. Yellow #5 had a theoretical max of 426-452 nm, the experimental max from 426-496
nm. Yellow #6 had a theoretical max of 421-484 nm, the experimental max was found to be 421490 nm. The reason for such variation in both the theoretical and experimental max could be due
to improper use or reading of the spectrophotometer results or general lab error.
In relation back to Beers Law, A= bc, where the absorbance varies linearly with both
the cell path length and the analyte concentration (Blaunch), the molar absorptivity was observed
after the calibration curves were created. In relation back to Table 4, the molar absorptivity for
Red #40 was 16150. In Group 1, a molar absorptivity of 22041 and Group 3 had 12278. For

Yellow #5, a molar absorptivity of 13099 was found; Group 1 had a molar absorptivity of 12196
and Group 3 had 28530. For Yellow #6, the molar absorptivity found was 4235.1. In Group 4,
there was a molar absorptivity of 8498 and Groups 5&6 had 5330. There may have been
difference among the results because molar absorptivity varies with the wavelength of light used
in measurement (Blaunch). So, if groups used different wavelengths to find the results, the molar
absorptivity will vary.
By creating a calibration curve for each food dye, it was able to be determined the
concentration of food dye in each commercial food product. The first food product tested was
M&Ms, which contained both Red #40 and Yellow #5. The concentration of Red #40 in M&Ms
was found to be 0.000015 M at a maximum absorbance of 0.2. The concentration of Yellow #5
was also found to be 0.000015 M at a maximum absorbance of 0.27. Next, for the Tic-Tacs, the
concentration of Yellow #6 was found to be 0.000038 M at a maximum absorbance of 0.18.
Conclusion
From the results, it is obvious that spectrophotometry can determine concentrations of
possibly unsafe dyes. The concentrations found of Red #40, Yellow #5, and Yellow #6 in the
commercial food products can allow insight into what manufacturers might not be telling
consumers. There is plenty of evidence that shows food dyes may be harmful to customers.
Research Connection
A recent study by researchers at the Slovak University of Technology was published in
the journal Food and Chemical Toxicology finding that blue dye used in edible products might be
doing more to the human body than what was thought (Oaklander). Study co-author Jarmila
Hojerov, an associate professor at the Slovak University of Technology and president of the

Slovak Society of Cosmetology, along with her research team, studied two blue dyes: Patent
Blue and Brilliant Blue. Hojerov and her colleagues have shown that the dyes can actually enter
the bloodstream via the skin or through the digestive tract (Oaklander). In comparison to the
experiment performed on dyes Red #40, Yellow #5, and Yellow #6, similar evidence has shown
the harmful effects of commercial dyes.

10

Works Cited
Blaunch, David N. "Spectrophotometry." Beer's Law. N.p., n.d. Web. 20 Apr. 2015.
Curran, Laurel. "Food Dyes Linked to Cancer, ADHD, Allergies." Food Safety News. N.p., 07
July 2010. Web. 20 Apr. 2015.
Jacobson, Michael F., Ph. D., and Sarah Kobylewski. "Food Dyes: A Rainbow of Risks." Center
for Science in Public Interest (2010): n. pag. Web. 19 Apr. 2015.
Oaklander, Mandy. "New Fear about Food Dyes." Fox News. FOX News Network, 17 Jan. 2013.
Web. 26 Apr. 2015.
Sandi-Urena, Santiago, Adrian Villalta-Cerdas, Todd Gaitlan, and Vailiki Lykourinou. General
Chemistry I Lab Manual. Tampa: Pro-Copy, 2015. Print.