Food dye lab report 2nd submission
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Identifying and Replicating the
Concentration of Food Dyes in Purple
Powerade Through the Use of
Spectroscopy, Beer’s Law, and the
Dilution Formula
Chemistry 1065 Fall 2023
Alex Wu,
,
,
Lucas Harrison
Anna Dittrich
Katie Reimer
Abstract
This experiment aimed to determine the concentration of various food dyes in a purple
Powerade. In this experiment, FD&C Blue #1, FD&C Blue #2, FD&C Red #3, and FD&C Red
#40 at different dilutions, along with the purple Powerade, were tested with an Ocean Optics
spectrophotometer in order to compare and contrast the peak wavelength and absorbance levels
to those found in the purple Powerade. Through the use of a spectrophotometer, it was
determined that Blue #1(628nm) and Red #40(498.3nm) were used in the drink as the peak
wavelengths matched closely with those of the purple Powerade, which had red and blue
wavelengths 628nm and 494nm respectively. Calibration curves were then created to find the
concentration using Beer’s Law. The resulting calculations found a concentration of
8.75*10^-6M of Red #40 and 3.37*10^-6 concentration of Blue #1 in the drink. Using the
dilution formula, trivially, 4.86ml of Red#40, 8.43ml of Blue #1 and 36.7ml of distilled water are
needed to recreate the Powerade. The recreated Powerade was then retested and compared to the
original to verify accuracy.
Introduction
Color spectroscopy and spectrophotometry have become essential tools in many scientific
fields due to their versatility and ability to accurately determine the composition of the
atmosphere and other tissues and objects. A 2020 meta-analysis published in the journal
“Progress of Biomedical Engineering,” showed the applications of optical spectroscopy in
diagnosing the
in vivo
disease states of certain tissues without the need for invasive biopsies.
When light is delivered in a highly localized state on tissues, certain combinations of optical
processes occur
1
. Analysis of these optical processes can rapidly provide in-depth insight into the
current pathological state of the tissue
1
, leading to faster, less invasive diagnosis and more
favorable patient outcomes. The tools and applications in this meta-analysis are much more
advanced than those used in this paper; however, the fundamental theory and purpose are similar.
Color spectroscopy is particularly useful in astronomy and searching for extraterrestrial
life in the universe. Every chemical element or molecule produces a specific light spectrum;
color spectroscopy equipment allows for the identification and analysis of these wavelengths
from distant celestial bodies. NASA’s exoplanet exploration department uses a type of color
spectroscopy called transmission spectroscopy in order to find life on other planets. The
molecules, methane, oxygen, and water, are all signs of potential life on other planets
2
; each of
these molecules has its own unique color signature, giving astronomers an unmistakable sign of
potential life
2
. Color spectroscopy's versatile and easy application provides a fundamental basis
for discovering life on other planets and propels the understanding of the nature of the universe
forward.
The purpose of this experiment is to identify and replicate the concentration of food dyes
in an artificially colored beverage using a spectrophotometer, Beer’s Law, and the dilution
formula. Due to the nature of this experiment, a hypothesis cannot be provided; however, if the
procedure is pertinently followed, the concentration of dyes will be found, and the color of
Powerade will be replicated. The contents of this report will elucidate the the procedures
followed in the experiment, the calculations done to find the concentration of food dyes in the
beverage, and the final result of the experiment.
Experimental
An artificially colored beverage with 2 different food dyes was obtained. The beverage
used in this experiment was a purple Powerade. The Powerade contained red #40 and blue #1
dyes in order to create its light purple color. An Ocean Optics spectrophotometer was used in
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order to determine the wavelength and absorbance levels of the dyes in the Powerade. The
spectrophotometer was connected to a laptop with LoggerPro software installed. The
spectrophotometer was calibrated using a cuvette filled with DI water. After calibration, the
cuvette was cleaned, dried, and filled with the Powerade. The software was then run with the
Powerade until two peaks occurred on the screen. The data was recorded in a spreadsheet.
After calibration and running the drink through the spectrophotometer, 4 dyes, Red #40,
Blue #1, Blue #2, and Red #3 at a pre-dilution concentration of 9*10
-5
M were tested. 5 different
concentrations of each dye were created. For each dye, a 25ml volumetric flask was filled with
water and poured into a beaker, then 25ml of the dye was poured into the beaker in order to
create a 50% concentration. This concentration was then measured by filling a cuvette and
running it through the spectrophotometer and LoggerPro until a peak occurred, with the data
recorded into a spreadsheet. Then, 25ml 50% concentration was disposed of into the sink and
replaced with 25ml of DI water to create a 25% concentration. The steps were repeated until the
concentration got down to 3.125%.
After measuring the wavelength and absorbance values of each concentration, the =max()
function in google docs was used in order to find the peak wavelength and absorbance values of
each dilution. The peak values of the Red #40 and the Blue #1 dilutions were then plotted to
form a linear regression. A trivial manipulation of Beer’s Law leads to the concentration value of
the two dyes. A simple bijection of the dilution formula leads to the amount needed for each dye
in order to replicate the color of the beverage. The beverage was replicated using 4.86ml of Red
#40, 8.43ml of Blue #1, and 36.7ml of water. The replicated color was then tested in the
spectrometer and compared to the original color.
Data
Table 1: Peak wavelength and absorbance values of Powerade and food dyes
Sample Tested
Peak Absorbance(AU)
Wavelength (nm)
Purple Drink(Red peak)
0.39
628.
Purple Drink(Blue peak)
0.18
494.
Blue #1
1.1
628.
Blue #2
0.39
608.
Red #40
0.32
498.
Red #3
0.04
528.
The Powerade was tested to show its peak red and blue wavelength and absorbance levels. All
dyes were then tested at 5 different concentrations in order to determine the peak absorbance and
wavelength.
Graph #1: Absorbance vs Wavelength in Powerade
The figure above shows two peaks in the Powerade, indicating the presence of different dyes in
the Powerade.
Graph #2: Concentration Curve of Red #40
The table above shows the linear regression of Red #40’s absorbance at its average wavelength
of all the dilutions of Red #40. The linear regression equation for this curve was
Y=2223X+0.09928.
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Graph #3: Concentration curve of Blue #1
The table above shows the linear regression of Blue #1’s absorbance at its average wavelength of
all the dilutions of Blue #1. The equation for this graph was Y= 110000364X.
Equation 1: Beer’s Law
A = (εl)(C)
Where:
In this equation, A is equal to absorbance, εl is equal to calibration curve slope, and C is equal
to concentration. Trivially, C = A/εl. This equation allows for the calculation of the
concentration of the red and blue dyes in the Powerade.
Equation 2: Dilution formula
C
1
V
1
=C
2
V
2
Where:
In this equation, C
1
is equal to the concentration of dye in Powerade, V
1
is equal to the volume of
Powerade, C
2
is equal to the concentration of dye used in replication Powerade V
2
is equal to the
volume of replicated Powerade. From Beer’s law, C
1
= A/εl. V
1
, V
2
are given. Trivally, C
2
=
((A/εl)(V
1
))/V
2
. This equation allows for the calculation of the concentration needed to recreate
the solution.
Discussion
A UV/vis spectrophotometer was used on a cuvette filled with Powerade to determine the
Powerade's wavelength and absorbance peaks. This testing showed that the Powerade had a red
peak at 628nm and a blue wavelength of 494nm, leading to the conclusion that Blue #1 and Red
#40 were the dyes used in the drink as their wavelengths(628nm and 498nm respectively) were
nearly identical to those in the Powerade. The peak values in each graph are able to be measured
as each wavelength has a specific energy level associated with it, meaning that when light hits
molecules, a wavelength with enough energy will be absorbed by the electrons of a molecule,
causing electrons to be promoted to a higher energy level. The graphs show this as the Y axis
shows the absorbance value, meaning that when wavelengths with enough energy to promote an
electron are absorbed, the graph spikes up, thereby showing the peak absorbance and wavelength
values. The accuracy of these peak points is of high importance in this lab as they ensure the
accuracy of the concentration values.
The concentration curves were created by plotting and running a linear regression on
each concentration's peak absorbance and wavelength values for the two dyes. The specific
molar concentrations of these two dyes were 4.5E^-5mol/L, 2.3E^-5mol/L, 1.1E^-5mol/L,
6.25E^-6 mol/L, and 2.8E^-6mol/L. The slope of these graphs allows for the concentration of the
two dyes to be calculated through Beer’s Law. In this experiment, the extinction coefficient is the
slope of the concentration curves. The value of the extinction coefficient in this experiment for
Red #40 was 22230molcm, and the extinction coefficient for Blue #1 was 110000364molcm.
The absorbance values for the dyes were determined by finding the corresponding absorbance
values. These values were 0.32AU for Red #40 and 1.1AU for Blue #1. Knowing these values,
an elementary application of algebra leads us to concentration values of 3.37E-6mol/L of Blue
#1 in the drink and a concentration of 8.75E-6mol/L of Red #40.
After finding the concentration values through Beer’s law, 4.86ml of Red #40, 8.43ml of
Blue #1 and 36.7ml of DI water were used in order to recreate the Powerade. While the
calculations that were made were all accurate, the visual appearance of the recreated drink was
visually a lot bluer than the original drink. This error can be attributed to the other chemicals
present in the Powerade; these chemicals could change the drink's color by clouding up the
sample, making it seem lighter or darker, affecting the spectra of the Powerade.
Other errors during this experiment can be attributed to the spectrophotometer's heat or
the cuvettes' quality and handling. In this experiment, the spectrophotometers were run for very
long periods of time, potentially causing the system to heat up, so when cuvettes were inserted
into the system, they could have absorbed this heat, subtly changing the properties of the
molecules in the solution, leading to an inaccurate reading. Improper handling or poor quality of
the cuvettes could skew results by potentially clouding up the outside of the cuvettes, preventing
the light from passing through easily and increasing the absorption reading as it would prevent
light from easily arriving at the sensor. This could lead to the spectrophotometer reading this as
absorption
3
, skewing the results.
Conclusion
This experiment aimed to determine the concentration of different dyes in an artificially
colored drink and replicate it. This was done using a spectrometer, Beer’s Law, and the dilution
formula. The spectrophotometer determined that the Powerade solution contained the dyes Blue
#1(498nm) and Red #40(628nm), as their wavelengths matched those seen in the
Powerade(494nm, 628nm) nearly perfectly. After plotting the concentration curves and applying
Beer’s Law, the concentration of the dyes in the Powerade was found to be 8.75E^-6M for Red
#40 and 3.37E^-6M for Blue #1. This information created a solution with 4.86ml of Red #40,
8.43ml of Blue #1 and 36.7ml of water. The sample solution was similar but not completely
identical to the original Powerade. This discrepancy could be caused by the other chemicals
present in the Powerade, heating up of the spectrophotometer, improper handling or the poor
quality of the cuvettes used.
This experiment can be expanded upon by using a different type of spectroscopy, such as
infrared spectroscopy. UV/Vis spectroscopy uses much higher energy and smaller wavelengths in
order to cause electron transitions in orbitals
3
, whereas infrared spectroscopy uses much lower
energy and longer wavelengths to affect protons within a molecule. The difference in the use of
these two techniques is that UV/Vis spectroscopy is often used for measuring concentrations of
known compounds in a solution, whereas infrared spectroscopy is useful in providing
information on the unknown compounds in the solution
4
. While UV/Vis spectroscopy is enough
for the scope of this lab, infrared spectroscopy can expand upon the scope by allowing for both
quantitative and qualitative understanding of solutions with unknown compounds.
This lab is important as it provides the fundamental basis for how UV/Vis spectroscopy
determines the various concentrations of compounds in a solution. UV/Vis spectroscopy is
extremely important in modern pharmaceutical quality control. With the mass production of
drugs such as ibuprofen, it is an efficient way to ensure that these drugs have the advertised
dosages
5
.
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References
(1) Kim, J. A.; Wales, D. J.; Yang, G.-Z. Optical Spectroscopy for in Vivo Medical
Diagnosis—a Review of the State of the Art and Future Perspectives.
Prog. Biomed. Eng.
2020
,
2
(4), 042001. https://doi.org/10.1088/2516-1091/abaaa3.
(2)
Why We Search | The Search For Life
. Exoplanet Exploration: Planets Beyond our Solar
System. https://exoplanets.nasa.gov/search-for-life/why-we-search (accessed 2023-11-28).
(3)
4.4: UV-Visible Spectroscopy
. Chemistry LibreTexts.
https://chem.libretexts.org/Bookshelves/Analytical_Chemistry/Physical_Methods_in_Chemi
stry_and_Nano_Science_(Barron)/04%3A_Chemical_Speciation/4.04%3A_UV-Visible_Spe
ctroscopy (accessed 2023-11-29).
(4)
Infrared Spectroscopy
. Chemistry LibreTexts.
https://chem.libretexts.org/Bookshelves/Physical_and_Theoretical_Chemistry_Textbook_Ma
ps/Supplemental_Modules_(Physical_and_Theoretical_Chemistry)/Spectroscopy/Vibrational
_Spectroscopy/Infrared_Spectroscopy/Infrared_Spectroscopy (accessed 2023-11-29).
(5) Al Ktash, M.; Stefanakis, M.; Boldrini, B.; Ostertag, E.; Brecht, M. Characterization of
Pharmaceutical Tablets Using UV Hyperspectral Imaging as a Rapid In-Line Analysis Tool.
Sensors (Basel)
2021
,
21
(13), 4436. https://doi.org/10.3390/s21134436.
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Which compound has the strongest binding to the μ receptor? Justify your answer. DJelsim Suppost 2² Tina, me 3 Course Hero' Rewards 2 OSHC Claims (Allian. 7 warded messa N Telsim Prepaid Pack [04933442 invoice and referral - Good morni Good news: you have new rewards o + bly Re-Further information Regi 8 Google recommends Chrome 24℃ 대체로 화창 A 9. 2023arrow_forward-OH -ова Ч OB₁ OTBS !.... OTBS ? n. мож .OTBS OTBI ова B₂ В Этски iPr Netarrow_forwardX Bb General Chemistry II CHEM 14 X G what does pka mean in chemis X ry II CHEM 1412, Richland College Dallas, Texas -us-east-1-prod-fleet01-xythos.content.blackboardcdn.com/blackboard.learn.xythos.prod/584b1d8497c84/143418394?X-Blackboard-S3-Bucket-blackbo F1 Q Practice Exercise 28. In the titration of 375 mL of 0.400 M HNO₂ (K₂ = 5.6 × 10-4) with 0.250 M NaOH, calculate the pH at each of these points: (a) before the addition of NaOH (b) after the addition of 375 mL of NaOH (c) at the equivalence point (d) after the addition of 650 mL of NaOH -0.³ 2 I W S 80 F3 $ ABAG 3 4 E 000 000 F4 D R F % 25/45 - 100% + 5 F5 T G Answer Key Chapter 14 - Chen X MacBook Air 6 F6 Y & 7 H F7 U * CO 8 DII F8 J in the titration of 375 ml of 0.4 X 9 1 L DD F9 K 0 0 B F10 L Parrow_forwardI'm not sure how to solve 115. I saw one solution that used the ka to solve it, but I don't understand how to find the ka to use or where they were able to get the ka from with the given information in the problem. So if you could please make this clear in the answer. Thank you for your help.arrow_forwardplease use the coverting table to answer #2arrow_forwardG neural crest tissue - Google Sear x A CHM 112 112-1 and 112-E1 A Presentation Session Student + 8 https://app.peardeck.com/student/twkkhzvar For quick access, place your favorites here on the favorites bar. Manage favorites now Peardeck Exercises Use #3. Nitol6) + 3 Fa(6)Z NF(1) + 3 HF(G) [1: 5,0 x10°n 0.10M Calculat K 2.0M 3.5 x 10°M Pear Deck Interactive Slide Students, draw anywhere on this slide! Do not remove this bar Slide 1/2arrow_forwardPlease answer all and post below the answer the source from where you got the answer thank you!arrow_forwardJust part b!!!arrow_forwardssignment/takeCovalent Activity.do?locator=assignment-take d... Communication s... 1.00 x 10-8 +1.71 x 10- -4 [References] Use the References to access important values if needed for this question. It is often necessary to do calculations using scientific notation when working chemistry problems. For practice, perform each of the following calculations. 2.37 x 104 +2.12 × 105 9.60 × 104 (1.00 QUT_Teamwork_... 28 7.00 × 10-5 0-8) (1.71 x 10-4) x 10 Submit Answer Retry Entire Group Aa Developing effecti... MacBook Pro Time managemen 3 more group attempts remaining Previous Nextarrow_forwardWhich of the following explains why a more concentrated dye solution has a higher absorbance? A. There are more dye molecules in the solution so more isotopes reach the detector. b. There are more isotopes in solution to emit photons. c. There are more dye molecules in solution to emit isotopes. d. There are more isotopes in solution to emit protons. e. There are more dye molecules in the solution to absorb photons.arrow_forwardB. Quantitative Spectroscopy Prepare two graphs, %T vs. Concentration and Absorbance vs. Concentration, using the data for the cobalt standards. The concentrations of Cobalt standards 2-4 can be calculated using simple dilution calculations. Concentration should be on the x-axis for each of these graphs. The first should clearly show some curvature while the second should be a linear curve. Using the absorbance graph you can determine the concentration of the unknown. Take the absorbance value for the unknown and find that value on the y-axis. Draw a horizontal line from that point on the y-axis to the plotted line. From the point where the horizontal line intersects the plotted line, draw a vertical line straight down to the x-axis. Record the value from the point where you hit the x-axis as the unknown concentration.arrow_forwardarrow_back_iosSEE MORE QUESTIONSarrow_forward_ios
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