Food Colorant Experiment

Davis Chap

Aidan Dietzler

Lab Section #019

Date: 2/14/22
Introduction
The experiment chosen will use absorption spectroscopy to allow you to find out
what type of dyes are used in beverages. We are using the dyes that are present and
constructing beer lambert plots. This will allow you to see which dyes and how
much dye are used in the creation of the original beverage. The graphs will be Beer-
Lambert plots as well as absorbance spectrum plots. Through absorption
spectroscopy, a sample of the name-brand beverage will be analyzed to determine
any dyes and the concentrations of the dyes that are present so that a generic
version can be produced.

The following equations will be used throughout the course of our experiment:
M1*V1=M2*V2
M= (mol of solute)/(L of solute)
E=(hc)/(wavelength)

Through this experiment, you will be identifying the relationship between

concentration and light absorption and utilizing that to develop a system for
determining the exact concentrations of any dyes that are present in the name-
brand beverage. Imagine a solution in a cuvette. When light passes through the
cuvette some of that light is absorbed in the species in the solution (atom, ion, or
molecule). This will result in a lesser amount of light leaving the solution than
originally entered. A device called a spectrophotometer will be used to record the
absorbance spectrum of the solutions we will be testing. The spectrophotometer
will record the quantity of light that reaches the other side as well as the wavelength
and amount of light absorbed by the sample. The wavelength that is correspondent
to the maximum absorbance band is known as the lambda max. The lambda max is
where all absorbance values should be recorded. We will be recording three known
concentrations of each dye and recording an absorbance spectrum for each. You will
then record the absorbance values at the lambda max. The lambda max can be
determined for a single spectrum but must be used for all other absorbance values
once determined. The absorbances values that you have recorded will then be
plotted versus the solution concentrations creating a Beer-Lambert Plot for each
dye. A linear regression analysis will be used on the plot. The equation and R^2
values for the linear fit should be shown on the plot. Using the equation from the
Beer-Lambert plot you can plug in the absorbance value for “y” and solve for the
concentration “x”. This will allow you to see where the name-brand solution’s
absorbance falls on the line locating the corresponding concentration.

Experimental
Sample of Name Brand Beverage
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)

Obtain cuvettes, four sheets of parafilm, 10 mL volumetric flask, cuvettes, beral

pipettes, and all of the previously listed solutions from the stockroom.
Prepare three different concentrations for each of the solutions, low (.05 mol of
solution to water), medium (.1 mol of solution to water), and high (.15 mol of
solution to water). Turn on your MeasureNet station and select Spectroscopy and
then F2 Absorption. Using a beral pipette, put one concentration of a solution into
one of the cuvettes, filled 75% of the way. Similar to the last step, add the sample of
a name-brand beverage to another cuvette, filled 75% of the way. Zero the
spectrometer by holding a light block cuvette to the sample holder and pressing the
‘zero’ button on your MeasureNet. You may use a DI water cuvette as your reference
during the spectroscopy. Take this absorption spectroscopy first and press the “R”
button on the MeasureNet. Repeat this process for each concentration of each
solution and save them under a file name. (e.g. 1001, 1002, 1003, etc.) The results
will be recorded in a graph of wavelength and absorption.

Results

The formula used is:

absorbance= wavelength*pathlength*concentration

We will use the following equation in order to obtain our different concentrations
for each dye:
M1V1=M2V2

Blue Dye Concentrations:

Show calculations for how you found each standard solution below:

Concentration Solution Water

0.00000675M 10ml 0ml

0.00000500M 7.4ml 2.6ml

0.00000250M 3.7ml 6.3ml

Red Dye Concentrations:

Concentration Solution Water

0.00004M 10ml 0ml

0.00003M 7.5ml 2.5ml

0.00002M 5ml 5ml

Yellow Dye Concentrations:

Concentration Solution Water

0.00004M 10ml 0ml

0.00003M 7.5ml 2.5ml

0.00002M 5ml 5ml

The absorbance spectrum of each solution will be graphed and should look similar
to the following samples below:
We will compare each of the solution’s absorption spectrum graphs containing the
concentration of each solution to the name-brand beverage solution and determine
what dyes the beverage contains as well as each of their concentrations through the
beer lambert plot.

Concentration Absorption

Bibliography

Burand, M. W. (2014). Food Colorant Specialist Position at YUME Food Products Inc.
Self-Directed Experiment [PDF]. Corvallis: Oregon State University.