Adamson University

College of Engineering

Chemical Engineering Department

Laboratory Report

Spectrophotometry
Experiment 1

Experiment run on: February 13, 2021

Submitted by:

Andallo, Jan Arvee V. 201912478

Austria, Jeremy Kyle Edson A. 201914365

Castro, Sean Carlo S.A. 201914624

Chua, Ma. Angelica B. 201914283

Reyes, Joserie Joice P. 201914413

Group 7

Submitted to:

Pinky Joy Janaban

Physical Chemistry for Engineers 1 Laboratory

51010

February 17, 2021

 1

ABSTRACT

Spectrophotometry is a technique for determining how much light is

absorbed by a chemical substance by measuring the amount of transmittance and
absorbance as the light beam moves through the sample solution (Chemistry
LibreText, 2020). The visible spectrum of red and blue dyes were used in this
experiment, at a concentration of 20 parts per million (ppm), to determine their
absorbance at various wavelengths. The spectrophotometer can only measure
wavelengths from 400 nm up to 680 nm where any wavelengths beyond and
below makes it not within range and gave us the maximum absorbance of 0.996
AU at 520 nm for the red dye and 0.886 AU at 620 nm for the blue dye. A high
percentage error occurred in the analysis of theoretical values for the absorbance
of red dye, 7.4% at concentrations 50 ppm and 100 ppm, and blue dye, 11.4% at
concentrations 10 ppm and 80 ppm. Concentration and path length is directly
proportional to the absorbance in Beer-Lambert’s Law.and 100 ppm, and blue
dye, 11.4% at concentrations 10 ppm and 80 ppm.
Keywords: absorbance, Beer’s Law, Spectrophotometry, transmittance,
wavelength

I. INTRODUCTION

Spectrophotometry is a method that measures the amount of light that

is absorbed in a chemical substance. This method is performed by an
instrument called a spectrophotometer which allows the observer to measure
the intensity of light as function of wavelength. It is commonly performed to
determine and analyze the presence of other chemical substances from the
sample.

This experiment can be beneficial for scientists and chemists observing

the chemical samples and determining the presence of an unknown chemical
compound from the collective medium. Several uses of spectroscopy include
determination of concentration of substances present, detection of impurities,
characterization of embedded proteins and functional groups in organic
 2

compounds, analysis of respiratory gas in hospitals, etc. Spectrophotometry is

indeed a valuable method for every researcher within the field of sciences and
engineering.

In this experiment, we will explore the fundamental principle that

applies concepts present in a theory named Beer – Lambert’s Law. This theory
defines the relationship concerning how an absorbing medium absorbs the
radiant energy project by the spectrophotometry. This experiment intends to
give insights and determine the following:

 To identify the different colors that can be perceived at different

wavelengths
 To determine the extent absorption from a spectrum of a solution with
the use of spectrophotometer
 To prepare Beer’s Law plot and determine the concentration of an
unknown solution through this plot.

II. THEORETICAL BACKGROUND

Every chemical substance is capable of absorbing, transmitting and

reflecting light of various wavelengths over the electromagnetic spectrum. To
determine the absorbance and transmittance of a chemical sample or solution,
spectrophotometry is usually the analytical technique or method used. This is
done through the use of a spectrophotometer, an instrument that passes a beam
of light through the sample and measures the light intensity absorbed. With
this, the concentrations of the substance can also be measured. (Chemistry
LibreText, 2020)

The concept behind the absorbance and reflection of light can easily be
determined and observed by the unaided eye. In a lit up surrounding, the colors
that the human eyes receive are constantly the wavelengths of light that are
reflected. In turn, colors from an object that are invisible to the eye are
 3

wavelengths of light that are absorbed. Each color has its corresponding
wavelengths in the electromagnetic spectrum. Longer wavelengths lean
toward the warmer colors of the spectrum like red and orange, while shorter
wavelengths lean toward the cooler colors of the spectrum like violet and blue.
Black absorbs all wavelengths while white reflects all wavelengths.

The spectrophotometer has two main components: the spectrometer

and the photometer, with the sample placed in between. The spectrometer
produces the desired wavelength while the photometer detects and measures
the intensity of light, converting it into a digital display. In a short overview of
this mechanism, the light from a lamp is separated into its wavelength
components through a prism and a specific wavelength is chosen to travel
through the sample. The amount of light that completely passes through the
sample is transmittance. Its opposite is absorbance which is the amount of light
that is absorbed (Chemistry LibreText, 2020). Thus, they are indirectly
proportional: the more the absorbance, the lesser the transmittance.

Two major factors affecting the absorbance and transmittance of light

is the concentration of the solution and the path length that light travels through
it. This is expressed through the Beer-Lambert Law which states that
absorbance has a directly proportional relationship to the concentration and
path length. When light passes through a highly concentrated solution, the
amount of molecules hinder its transmittance, increasing its absorbance. In the
same way, a longer path length makes light pass through more air molecules
which also increases the absorbance. (AAT Bioquest, 2020)
 4

BEER – LAMBERT’S LAW:

Abs = εlc

Where:
Abs = Absorbance
ε = Molar Absorptivity coefficient
l = Path length
c = Concentration

𝐼 𝑻 𝐼𝑜
T = 100 (𝐼𝑜 ) ; A = -log10 (
100
) = log10 ( 𝐼 )

Where:
T = Transmittance
A = Absorbance
I = Intensity of incident light
Io = Intensity of transmitted light

III. PROCEDURE

In this experiment, the researchers have identified the colors at different

wavelengths in the spectrum and determined the relationship of absorption with
the use of a spectrophotometric simulation. The following simulation has been
used to demonstrate the relationships between the components of Beer-Lambert’s
Law consisting of wavelength, concentration, and absorbance. Here are the
following procedures that are undertaken to complete this experiment:

a. Color of Visible Radiation. This part of the experiment aims to

determine the color that is perceived by the naked eye in different
wavelengths within the spectrum. With the aid of previous literature by
 5

other researchers, we have determined the color present at varying

wavelengths. Furthermore, we have determined how the color emitted
or perceived can change with a corresponding change in wavelength.
With this inference, we can estimate the theoretical wavelengths of
several chemicals that project a particular type of color.
b. Visible Spectrum of Red and Blue Dye. In this part of the experiment,
the researcher will determine the behavior of absorbance in varying
wavelengths. Here are the following steps that are executed to collect
and analyze the data:
1. Turn on the spectrophotometer to start the operation.
2. Click the mode button until the option presents ABS
(Absorbance).
3. Click the upward red button in order to get a dye sample of a
specific concentration. In this experiment 20 ppm of red dye
and 10ppm of blue dye were obtained and transferred from the
bottle into a cuvette.
4. Click on the cuvette in position 0 or the first cuvette in the
rack, and then bring it into the cell compartment. (An initial
reading will be read which displays at what wavelength the
spectrophotometer is currently set at).
5. Set the desired wavelength by clicking on the set wavelength
button below the spectrophotometer. The range is set to 400-
680 nm. Start with 400 nm then click the OK button.
6. Click the auto zero button in order to reset the reading.
7. Remove the cuvette in the cell compartment and click on the
cuvette on position #1 or where the 20 ppm red dye was placed
and put it inside the cell compartment. Do the same for the
blue dye.
8. Record the obtained absorbance value.
 6

9. Repeat steps 5 to 8 with the wavelength changed in increments

of 10 until 680 nm is reached. (The scan button can also be
used in order to obtain the absorbance values)
10. Graph the values obtained.
c. Determination of Concentration using Absorbance Values (For
Red and Blue Dye). This part explores how to determine the
concentration from the assumed absorbance values. The empirical
formula will be determined from initial experimentation. Here are the
following procedures that is done to complete the data:
1. Set the mode to Transmittance, click on the mode button to
change it. The cell compartment should be empty and closed
and click on the left knob, to turn on the instrument.
2. Operate the left knob to bring the Transmittance reading to 0.0.
3. Operate the large knob on the upper top deck of the instrument
to set the desired wavelength. Alternatively, click on set
Wavelength and type the desired value.
4. Click the mode button to change the mode to Absorbance.
5. Click the reference cuvette in position 0 to put it on the cell
compartment. Operate the right knob to bring the Absorbance
reading to 0.000. More easily, this can be done by clicking the
AutoZero button. The instrument must be zeroed with the
reference solution at each wavelength.
6. Click the cell compartment to remove the reference cuvette.
Click on one of the other cuvettes to put it on the cell
compartment. Record the Absorbance reading and the
Wavelength.
7. Repeat this step to measure the absorbance for each of the
cuvettes with different concentrations at this wavelength.
8. When done executing the experiment for the first dye, repeat
steps A to G for the second dye.
 7

d. Data Analysis. For this part, the researcher will use the initial
experimental results from part C for the determination of concentration
using absorbance values will be plotted into a linear regression curve
to obtain an empirical equation. This equation will be true for all the
values in the particular graph based on data points gathered from the
experiment. From this method, we can determine any data with a given
assumed value. Furthermore, to ensure the accuracy of the simulation
used for this experiment, we will test the comparison of the theoretical
and experimental values to determine for lapses or errors in the results
of the experiment.
 8

IV. RESULTS AND DISCUSSION

A. VISIBLE SPECTRUM OF RED DYE (CONCENTRATION, 20ppm)

WAVELENGTH ABSORBANCE
(nm) (AU)
400 0
410 0.001
420 0.004
430 0.01
440 0.028
450 0.064
460 0.119
470 0.178
480 0.242
490 0.346
500 0.541
520 0.996
530 0.927 Mean 0.17692
540 0.547 Median 0.029
550 0.208 Mode 0.001
560 0.076
570 0.043
580 0.029
590 0.014
600 0.008
620 0.001
640 0.001
650 0.004
670 0.013
680 0.023
Table 1. Analysis of the Visible Spectrum of Red Dye
 9

Figure 1. Graphical Representation of the Absorbance of Red Dye

The spectrophotometer used for this experiment only covers wavelengths of

light ranging from 400 nm to 680 nm. Any other wavelengths outside this range are
excluded. In addition, only two samples of dyes, red and blue, were used for this
spectrophotometry. With a constant variable of concentration at 20 ppm, every 10
nm or 20 nm from 400 nm until 680 nm, the absorbance is recorded in a table in
absorbance units (AU).
For the first sample of red dye, the data gathered is tabularized and
accompanied by its calculated mean, median, and mode as shown in Table 1. It has
a mean or average absorbance of 0.17692 AU and a median of 0.029 AU at 580 nm.
Its mode, the most occurring absorbance recorded, is 0.001 AU at the wavelengths
of 410 nm, 620 nm and 640 nm.
Following the table is Figure 1, a graphical representation of the red dye’s
absorbance based on the Table 1. The wavelength and absorbance are plotted against
each other wherein the independent variable is wavelength on the x-axis and the
dependent variable is absorbance on the y-axis. The graph shows positive kurtosis
and a leptokurtic distribution of the data meaning that most of the values are located
in the tails. Upon analysis, the graph illustrates the red dye’s behavior towards these
 10

wavelengths of light. It shows more absorbance in the range 450 nm to 560 nm and
displays a maximum 0.996 AU at 520 nm. It also shows a minimum absorbance of
0 AU at 400 nm. This means that the red dye is able to absorb colors ranging from
blue to green on the electromagnetic spectrum end of shorter wavelengths.
 11

b. VISIBLE SPECTRUM OF BLUE DYE (CONCENTRATION, 20ppm)

WAVELENGTH ABSORBANCE
(nm) (AU)
Not within the
375
range
400 0.126
410 0.112
420 0.077
430 0.044
440 0.019
450 0.007
460 0.002
470 0.007
480 0
490 0.001 Mean 0.18464
500 0.002 Median 0.065
520 0.014 Mode 0.007, 0.002
530 0.025
540 0.039
550 0.065
560 0.12
570 0.211
580 0.311
590 0.397
600 0.504
620 0.886
640 0.858
650 0.6
670 0.141
680 0.048

Table 2. Analysis of the Visible Spectrum of Blue Dye

 12

Figure 2. Graphical Representation of the Absorbance of Blue Dye

The same procedure was performed for the second sample of blue dye and
recorded similarly as shown in Table 2. This set of data has an average absorbance
(mean) of 0.18464 A, a median of 0.065 AU at 550 nm, and a repetitive occurrence
(mode) of absorbance 0.002 AU at wavelengths 460 nm and 500 nm and absorbance
0.007 AU at wavelengths 450 nm and 470 nm.
Following this table is Figure 2, a graphical representation of the blue dye’s
absorbance based on the Table 2. The plotting of independent and dependent
variables are the same as Figure 1 which is the wavelength (x-axis) versus its
absorbance (y-axis). The graph also shows a positive kurtosis and leptokurtic
distribution with most of the data along its tails. Based on the plot’s trend, it can be
inferred that the blue dye becomes more absorbent in the ranges 550 nm to 680 nm,
and more transmittant in other wavelengths. According to the data, it reaches a peak
or maximum absorbance of 0.886 AU at 620 nm, but it can also be deduced from
the graph that the actual peak lies in between 620 nm to 640 nm. The minimum or
least absorbance is found at 480 nm where there is 0 AU or no absorbance at all.
 13

This means that the blue dye is able to absorb colors ranging from green to red on
the electromagnetic spectrum end of longer wavelengths.

c. DETERMINATION OF CONCENTRATION USING ABSORBANCE

VALUES

WAVELENGTH, 520 nm
RED DYE
CONCENTRATION (PPM) ABSORBANCE
0 0
10 1.073
50 1.074
80 1.073
100 1.074

Mean 0.8588
Median 1.073
Mode 1.073, 1.074
Margin of Error ± 0.001

Table 3.1 Analysis of Absorbance using Concentration Values of Red Dye

In this part of the experiment, the wavelength of light is kept as the constant
variable. The wavelength is set to 520 nm where the red dye had its maximum
absorbance. This time the absorbance is measured against various concentrations
namely, 0 ppm,10 ppm, 50 ppm, 80 ppm and 100 ppm. Also, the margin of error is
added to the table of mean, median and mode. This helps in perceiving the slight
variances of absorbance values between the concentrations, also in contrast to the
known absorbance of red dye at 20 ppm. It can be seen that the average absorbance
is 0.8588 AU with a median of 1.073 AU and mode of 1.073 AU and 1.074 AU. It
also contains a margin of error of ± 0.001. Based on the Beer’s Law, absorbance
must increase as the concentration increases. This, however, is not visible in the
information that Table 3.1 provides; it does not align with the Beer’s Law.
 14

Figure 3. Beer’s Law Plot for the Concentration of Red Dye

Table 3.2. Analysis of Theoretical Values for the Absorbance of Red Dye
Empirical equation: Abs = 0.0136(Conc)
Absorbance Absorbance Percentage Error
Concentration
(Theoretical) (Experimental) (% error)
0 0 0 undefined
10 0.136 1.073 7.3
50 0.68 1.074 7.4
80 1.088 1.073 7.3
100 1.36 1.074 7.4

The Beer’s Law Plot is shown in Figure 3 where a linear regression is

made to show the proper direct and linear relationship of absorbance and
concentration. This led to the finding of the empirical equation, Abs =
0.0136(Concentration), to produce theoretical values based on the experimental
data. Starting from (0,0), only one point of the plotted data came close to the
linear function. The other three values significantly deviate from the function,
already visibly implying a large percentage error from the experimental data.
 15

This is ascertained in Table 3.2 in which the theoretical data,

experimental data and percentage error are tabularized. Unlike the experimental
data that fluctuates with a ± 0.001 margin of error, the theoretical data of
absorbance values over increasing concentration adheres to the Beer’s Law. It
increases as the concentration increases and is more likely to be situated close to
or around the linear function in Figure 3 when plotted. The percentage errors
calculated for the absorbance of red dye resulted in 7.3% and 7.4%.

WAVELENGTH, 620 nm
BLUE DYE
CONCENTRATION (PPM) ABSORBANCE
0 0
10 0.886
50 0.889
80 0.886
100 0.888

Mean 0.7098
Median 0.886
Mode 0.886
Margin of Error ± 0.003

Similar to the tabulated data for the red dye, the wavelength for the blue
dye is set as a constant variable at 620 nm where it had its maximum absorbance.
The same procedure is performed yielding an average absorbance of 0.7098 AU
with a median and mode of 0.886 AU. Its margin of error is ± 0.003 which is
slightly larger than the red dye’s in Table 3.1. Here, it is also visible that the
absorbance values remain consistent and do not adhere to Beer’s law as the
concentration increases.
 16

Figure 4. Beer’s Law Plot for the Concentration of Blue Dye

Empirical equation: Abs = 0.0112(Conc)

Absorbance Absorbance Percentage Error (%
Concentration (Theoretical) (Experimental) error)
0 0 0 #DIV/0!
10 0.112 0.886 11.4
50 0.56 0.889 11.1
80 0.896 0.886 11.4
100 1.12 0.888 11.2

Table 4.2. Analysis of Theoretical Values for the Absorbance of Blue Dye

Figure 4 is the Beer’s Law Plot for the blue dye where the tabularized
data is plotted and linear regression is applied to get the empirical equation,
Abs = 0.0112(Concentration). Only one point aligns with the linear function
like Figure 3 and the theoretical absorbance values are calculated. The
theoretical data exhibits the directly proportional relationship of absorbance
 17

and concentration and follows the Beer’s Law more accurately than the
experimental data. The information is arranged in Table 4.2 and it can be
seen that the percentage errors are 3% more than the percentage in the red
dye namely, 11.1%, 11.2% and 11.4%.
These large percentage errors can potentially be due to the
spectrophotometer simulation used for the experiment. Errors in the
simulation may have caused significant changes in the results. The
discrepancies present in the experiment proves a slight inaccuracy on the
reliability of the instrument or simulation used for this experiment.
 18

V. CONCLUSION AND RECOMMENDATIONS

Colors were seen, at different wavelengths, on a visible radiation
spectrum which is proven to be useful in estimating the wavelengths of
several chemicals that project a particular type of color. Longer wavelengths
showed warmer colors while shorter wavelengths leaned towards cooler
colors. Red and blue dyes tested in the experiment showed that the
relationship between absorbance and concentration is directly proportional
to each other. The presence of various wavelengths of light shone on
chemical substances can affect the value of the absorbance. Upon testing in
a constant wavelength, different concentrations gave almost the same
absorbance value with minimal margin of error but did not adhere to the
Beer’s Law causing a large percentage error. Overall, this experiment
successfully observed the principle of Beer’s Law which helped to determine
how the wavelengths, concentrations and absorbance values affect each other
when one variable is held constant. The Beer’s law was more evident in the
theoretical values in contrast to the experimental values.
It is highly recommended that for this type of experiment, the instructor
needs to provide a theoretical basis conducted by other researchers or
scientists in order to write laboratory reports faster and with accurate results.
Also, simulations are designed to imitate the real equipment in a real
laboratory setting, spectrophotometer, so the virtual equipment should be
able to avoid errors in any way.
 19

VI. LITERATURE CITED

AAT Bioquest. (2020, October 30). What factors will affect absorbance?
Retrieved from https://www.aatbio.com/resources/faq-frequently-
asked-questions/What-factors-will-affect-
absorbance#:%7E:text=The%20two%20main%20factors%20that,con
centration%2C%20the%20higher%20its%20absorbance.&text=This
%20increases%20the%20absorbance

Beer-Lambert Law. (n.d.). Retrieved from

http://life.nthu.edu.tw/%7Elabcjw/BioPhyChem/Spectroscopy/beersla
w.htm

Den, B. (2020, February 4). Spectrophotometer Instrumentation : Principle and

Applications. Biochemistry Den. Retrieved from
https://biochemden.com/spectrophotometer-instrumentation-
principle/

Derivation of Beer Lambert Law. (n.d.). Retrieved from

https://byjus.com/physics/derivation-of-beer-lambert-law/

Fadak A. & Mazahernasab R. (2013, June 24). Spectrophotometry : Instruments

& Applications. slideshare.net. Retrieved from
https://www.slideshare.net/rey216/spect

Libretexts. (2020, August 15). 2.1.5: Spectrophotometry. Chemistry LibreTexts.

https://chem.libretexts.org/Bookshelves/Physical_and_Theoretical_C
hemistry_Textbook_Maps/Supplemental_Modules_(Physical_and_Th
eoretical_Chemistry)/Kinetics/02%3A_Reaction_Rates/2.01%3A_Ex
perimental_Determination_of_Kinetics/2.1.05%3A_Spectrophotomet
ry

Walker, K. (2017, August 1). Spectrometer Technology and Applications.

AZoM.Com. https://www.azom.com/article.aspx?ArticleID=10245

Spectrophotometer Principle. (n.d.). Retrieved from

https://byjus.com/chemistry/spectrophotometer-principle/
 20

VII. NOMENCLATURE
In this experiment, the researchers have determined the relationship
between several factors that affect the absorbance and concentration of a sample.
Analyzing this experiment, we have utilized the Beer-Lambert equation which
was used to determine the given unknown. This following equation consists of
the following variables and terminology that are presented and defined in this
table below:

SYMBOL QUANTITY DEFINITION SI UNIT

Abs Absorbance Amount of light absorbed by AU

the solution

c Concentration Number of molecules per mol L-1

unit volume

Io Initial Light Intensity of incident light Unitless

Intensity upon the solution

I Light Intensity Intensity of transmitted light Unitless

leaving the solution

ε Molar Efficiency of the sample to L mol-1 cm-1

Absorptivity absorb light at a given
wavelength

l Path length Distance light travels cm

through the solution

T Transmittance Amount of light transmitted Unitless

by the solution
 21

APPENDICES
 22

APPENDIX A.1
RAW DATA
 23

APPENDIX A.1. I.

COLOR OF VISIBLE RADIATION

WAVELENGTH COLOR WAVELENGTH COLOR

400 Violet 650 Red

450 Blue 700 Red

500 Cyan 750 Red

550 Green 800 Red

600 Yellow
 24

APPENDIX A.1. II.

VISIBLE SPECTRUM OF RED DYE (CONCENTRATION, 20ppm)

WAVELENGTH ABSORBANCE WAVELENGTH ABSORBANCE WAVELENGTH ABSORBANCE

Not within
375 520 0.996 670 0.013
range

400 0.000 530 0.927 680 0.023

Not within
410 0.001 540 0.547 690
range

Not within
420 0.004 550 0.208 700
range

Not within
430 0.010 560 0.076 710
range

Not within
440 0.028 570 0.043 720
range

Not within
450 0.064 580 0.029 730
range

Not within
460 0.119 590 0.014 740
range

Not within
470 0.178 600 0.008 750
range

Not within
480 0.242 620 0.001 760
range

Not within
490 0.346 640 0.001 770
range

Not within
500 0.541 650 0.004 780
range
 25

APPENDIX A.1.III.

VISIBLE SPECTRUM OF BLUE DYE (CONCENTRATION, 20ppm)

WAVELENGTH ABSORBANCE WAVELENGTH ABSORBANCE WAVELENGTH ABSORBANCE

Not within
375 520 0.014 670 0.141
range

400 0.126 530 0.025 680 0.048

Not within
410 0.112 540 0.039 690
range

Not within
420 0.077 550 0.065 700
range

Not within
430 0.044 560 0.120 710
range

Not within
440 0.019 570 0.211 720
range

Not within
450 0.007 580 0.311 730
range

Not within
460 0.002 590 0.397 740
range

Not within
470 0.007 600 0.504 750
range

Not within
480 0.000 620 0.886 760
range

Not within
490 0.001 640 0.858 770
range

Not within
500 0.002 650 0.600 780
range
 26

APPENDIX A.1. IV.

DETERMINATION OF CONCENTRATION USING ABSORBANCE

VALUES

WAVELENGTH, 520 nm

RED DYE
CONCENTRATION (PPM) ABSORBANCE
0 0.000
10 1.073
50 1.074
80 1.073
100 1.074

WAVELENGTH, 620 nm

BLUE DYE
CONCENTRATION (PPM) ABSORBANCE
0 0.000
10 0.886
50 0.889
80 0.886
100 0.888
 27

APPENDIX A.2

ANALYSIS OF DATA AND SAMPLE CALCULATIONS

 28

APPENDIX A.2.V.

ANALYSIS OF THE DETERMINATION OF CONCENTRATION USING

ABSORBANCE VALUES

WAVELENGTH, 520 nm
RED DYE
CONCENTRATION (PPM) ABSORBANCE
0 0
10 1.073
50 1.074
80 1.073
100 1.074

Mean 0.8588
Median 1.073
Mode 1.073
Confidence Interval ± 0.001

Empirical equation:
Abs = 0.0136(Conc)
Percentage
Absorbance Absorbance Error
Concentration (Theoretical) (Experimental) (% error)
0 0 0 #DIV/0!
10 0.136 1.073 7.3
50 0.68 1.074 7.4
80 1.088 1.073 7.3
100 1.36 1.074 7.4
 29

SAMPLE CALCULATIONS

Concentration Values Percentage error

Formula:
Concentration = 0 % 𝑒𝑟𝑟𝑜𝑟
Abs = 0.0136(Conc) 𝑎𝑐𝑡𝑢𝑎𝑙 𝑣𝑎𝑙𝑢𝑒 − 𝑒𝑥𝑝𝑒𝑟𝑖𝑚𝑒𝑛𝑡𝑎𝑙 𝑣𝑎𝑙𝑢𝑒
Abs = 0.0136(0) =| | 𝑥 100
𝑒𝑥𝑝𝑒𝑟𝑖𝑚𝑒𝑛𝑡𝑎𝑙 𝑣𝑎𝑙𝑢𝑒
Abstheo = 0
Concentration = 0
Concentration = 10
Abs = 0.0136(Conc) 0−0
% 𝑒𝑟𝑟𝑜𝑟 = | | 𝑥 100
Abs = 0.0136(10) 0
Abstheo = 0.136 = undefined
Concentration = 50 Concentration = 10
Abs = 0.0136(Conc) 0.136 − 1.073
Abs = 0.0136(50) % 𝑒𝑟𝑟𝑜𝑟 = | | 𝑥 100
1.073
Abstheo = 0.68
=7.3%
Concentration = 80
Abs = 0.0136(Conc) Concentration = 50
Abs = 0.0136(80) 0.68 − 1.074
Abstheo = 1.088 % 𝑒𝑟𝑟𝑜𝑟 = | | 𝑥 100
1.074
Concentration = 100 =7.4%
Abs = 0.0136(Conc)
Abs = 0.0136(100) Concentration = 80
Abstheo = 1.36
1.088 − 1.073
% 𝑒𝑟𝑟𝑜𝑟 = | | 𝑥 100
1.073
=7.3%

Concentration = 100

1.36 − 1.074
% 𝑒𝑟𝑟𝑜𝑟 = | | 𝑥 100
1.074
=7.4%
 30

WAVELENGTH, 620 nm
BLUE DYE
CONCENTRATION (PPM) ABSORBANCE
0 0
10 0.886
50 0.889
80 0.886
100 0.888

Mean 0.7098
Median 0.886
Mode 0.886
Confidence Interval ± 0.003

Empirical equation
Abs = 0.0112(Conc)
Percentage
Absorbance Absorbance
Concentration Error (%
(Theoretical) (Experimental)
error)
0 0 0 undefined
10 0.112 0.886 11.4
50 0.56 0.889 11.1
80 0.896 0.886 11.4
100 1.12 0.888 11.2
 31

SAMPLE CALCULATIONS

Absorbance Values Percentage error

Formula:
Concentration = 0 % 𝑒𝑟𝑟𝑜𝑟
Abs = 0.0112(Conc) 𝑎𝑐𝑡𝑢𝑎𝑙 𝑣𝑎𝑙𝑢𝑒 − 𝑒𝑥𝑝𝑒𝑟𝑖𝑚𝑒𝑛𝑡𝑎𝑙 𝑣𝑎𝑙𝑢𝑒
Abs = 0.0112(0) =| | 𝑥 100
𝑒𝑥𝑝𝑒𝑟𝑖𝑚𝑒𝑛𝑡𝑎𝑙 𝑣𝑎𝑙𝑢𝑒
Abstheo = 0

Concentration = 10 Concentration = 0
Abs = 0.0112(Conc)
Abs = 0.0112(10) 0−0
% 𝑒𝑟𝑟𝑜𝑟 = | | 𝑥 100
Abstheo = 0.112 0

Concentration = 50 = undefined
Abs = 0.0112(Conc) Concentration = 10
Abs = 0.0112(50) 0.112 − 0.886
Abstheo = 0.56 % 𝑒𝑟𝑟𝑜𝑟 = | | 𝑥 100
0.886
Concentration = 80 =11.4%
Abs = 0.0112(Conc)
Abs = 0.0112(80) Concentration = 50
Abstheo = 0.896 0.56 − 0.889
% 𝑒𝑟𝑟𝑜𝑟 = | | 𝑥 100
Concentration = 100 0.889
Abs = 0.0112(Conc) =11.1%
Abs = 0.0112(100)
Abstheo = 1.12 Concentration = 80

0.896 − 0.886
% 𝑒𝑟𝑟𝑜𝑟 = | | 𝑥 100
0.886
=11.4%

Concentration = 100

1.12 − 0.888
% 𝑒𝑟𝑟𝑜𝑟 = | | 𝑥 100
0.888
=11.2%
 32

APPENDIX A.3

SIMILARITY REPORT
33