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Behavior of Different Color Dyes When Diluted

Author’s Name: Aiden Dunn

General Chemistry for Engineers Lab

Section Number: 5

Instructor's name: Zachary Magnuson

Due: October 28, 2022

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Introduction:

Spectrophotometry can be used to determine the distance and age of specific stars. This is done

by studying how radiation and matter interact with each other in order to find objects structure

and its molecules behavior. For example, in the article “Hubble Space Telescope

Spectrophotometry and Models for Solar Analogs”, the author discusses how the Hubble Space

Telescope uses spectrophotometry in order to measure stars' distances from Earth. This report

covers the principles of spectrophotometry for chemical analysis and how to explain the

interaction between UV-Visible radiation and molecules in commercial products. In this

experiment, several different dyes will be measured for their dilution and concentration of each

dilution to find their specific absorbance by using a spectrophotometer. By creating a calibration

plot, the calibration curve will then be used to correlate it to Beer’s Law to find the unknown

concentration. By using Beer’s Law, the absorbance capacity of a substance will be found.

Experimental Methods:

Part one:

The group first chose three different color dyes to conduct parallel dilutions for all dyes. The

group created a stock solution that had an absorbance rating around .95. The group then

completed a parallel dilution starting from 8 mL of the stock solution to 2 mL of di water, 6 mL

of stock solution to 4 mL of di water and so on until it the last dilution was 2 mL of stock

solution to 8 mL of di water. The rating of absorbance was then measured using the

spectrophotometer for each different level of dilution. Between each dilution creation, every

flask and mixing tool was properly rinsed with di water to ensure no cross contamination. The

group then repeated the same steps for the other two dyes. The group then created the calibration
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curves on Excel from all of the dyes. This helped to obtain the concentration of the dyes and the

R^2 of the curve.

Part two:

The group brought in three different commercial products with the same color dyes as the ones

previously measured in the lab. The group then diluted the three different commercial products,

if needed, just enough to have an absorbance level around .95 at the same wavelength at which

the previous three dyes were measured at. The absorbance rate was measured by using the

spectrophotometer. The group then found the concentration of each commercial product by using

y=mx+b. Where y=absorbance, m=the slope, x=concentration, and b=the y-value on the

calibration curve.

Results:

Table: 1. Commercial Food Products Analysis

Absorbance Wavelength (nm) Concentration (M)

Green #3: Listerine 0.249 618.4 0.00026

Yellow #6: orange 0.842 437.5 0.000429

Mio

Blue #1: blue 0.535 618.4 0.000246

Gatorade
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Table 2: Dye dilution with different lambda max

Commercial Concentration V2 (mL) M1 (stock, M) V1 (mL)

Products (M2)

Orange Mio 0.000291 51 mL 0.021879 1 mL

Table 3: Blue Dye #1

concentration absorption

0.0005 0.992

0.0004 0.798

0.0003 0.677

0.0002 0.465

0.0001 0.238
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Table 4: Green Dye #3

concentration absorption

0.0005 0.947

0.0004 0.845

0.0003 0.64

0.0002 0.472

0.0001 0.215

Table 5: Yellow Dye #5

concentration absorption

0.0005 0.969

0.0004 0.767

0.0003 0.606

0.0002 0.393

0.0001 0.179

Graph 1: Blue Dye #1

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Table 2: Green Dye #3

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Table 3: Yellow Dye #5

Calculations:

y=mx+b
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Listerine: 0.249=1837x+0.727=0.00026 x=0.00026 M

Orange Mio: 0.842=1954x+0.0034=0.000429 x=0.000429 M

Blue Gatorade: 0.535=1841x+0.0817 x=0.000246 M

Discussion:

The linear equation for blue dye #1 is y=1841x+0.0817, for green dye #3 is y=1837x+0.0727,

and for yellow dye #5 the linear equation is y=1954x+0.0034. The y variable is the absorbance

value, the x variable is the unknown concentration, the m variable is the slope of the best fit line,

and the b variable is the y intercept. The R-squared values for the graphs are blue dye #1=.9902,

green dye #3=.9815, and yellow dye #5=.9977. The R-squared numbers mean how close the

values are to the line of best fit and is between the range of 0 and 1 meaning 1 is a perfect fit to

the line of best fit. For instance, in the research article “Adjusted R-squared Type Measure for

Exponential Dispersion Models’, it says how the R-squared value is valuable variable when

considering the validity of a group of specific values. The linear equation relates to Beer’s Law

because they both include the absorbance and concentration. The concentration can be found

when using Beer’s Law by using the absorbance found for the unknown and the slope of the best

fit line, to find the concentration of the unknown solution. Some potential errors that can occur

when conducting this experiment are not properly rinsing every piece of glassware, which could

cause cross contamination of the different rates of dilution. Another error that could happen is

when using the spectrophotometer, someone could contaminate the outside of the cuvette with

another type of material that could cause the spectrophotometer to get a miss read. While most of

the data values are very accurate, there could still possibly be some form of human errors. Also,

in the article “Using Human Error Information for Error Prevention”, it is discussed that when

the students have a better understanding of how human errors occur, the number of human errors
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when conducting an experiment can be drastically reduced. This is why if I was to repeat this

experiment, I would conduct multiple trials with the same dye in order to get more accurate data

values.

Conclusion:

In conclusion, the results of the experiment are very accurate because of how the R-squared

value is for all three graphs. This can be seen in the discussion where it states that the R-squared

value is only accurate when the value is close to 1. Another reason why the experiment was a

success is because all the calculations seem to line up to the corresponding values that the group

was looking for. For example, the absorption values in tables 3 through 5 are linearly decreasing

from a value starting around .95 and ending around a value of .2. Also, the linear equation for all

three graphs matches up with the three commercial products that the group brought in to be

measured. The results show that each commercial product was correctly measured and calculated

as seen in the calculations for the M value.

References:

Bohlin, R. C. Hubble Space Telescope spectrophotometry and models for solar analogs.

https://www.osti.gov/biblio/21301202-hubble-space-telescope-spectrophotometry-models-

solar-analogs (accessed Oct 28, 2022).

Hu, W.; Carver, J. C.; Anu, V.; Walia, G. S.; Bradshaw, G. L. Using human error information for

error prevention - empirical software engineering.

https://link.springer.com/article/10.1007/s10664-018-9623-8 (accessed Oct 28, 2022).

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Ricci, L. Adjusted R-squared type measure for exponential dispersion models.

https://www.sciencedirect.com/science/article/pii/S0167715210001288 (accessed Oct 28,

2022).