STCH 205L Qualitative with Quantitative Chemistry Laboratory II

Date performed: 02/7/24

Date submitted: 02/20/24

Determining the wavelength of the maximum absorbance of

Bromocresol Green (BCG)
Justine Andrew P. Matias
Abstract
The objective of this study is to understand the principles of spectrophotometry and to determine
the maximum wavelength and absorbance of bromocresol green (BCG) using a UV-Vis
spectrophotometer. The standard BCG solution was prepared using 30mL of 0.0001 M stock
solution. Using a UV-Vis spectrophotometer, the 𝜆max was determined and a series of dilution
was prepared using a test tube, having a ratio of BCG: H2O. The λmax of BCG was determined
to be 610 nm, which is consistent with the values of the literature. Furthermore, the absorbance
of the dilution series of BCG solutions decreased as the concentration decreased, revealing a
clear relationship between the two variables. the relationship between absorbance and
concentration was studied, which confirmed the validity of Beer-Lambert's Law.

Keywords: Spectrophotometry, bromocresol green, UV-Vis spectrophotometer, Beer-Lambert’s

Law,

Introduction
Spectroscopy is a broad area of Chemistry
that deals with the study of the interaction
between light and matter. Electromagnetic
radiation is an energy that can be in the form
of light, heat, radiation, etc. that includes
radio waves, microwaves, infrared, visible
light, ultraviolet light, x-ray, and gamma
rays. Upon the interaction of EM waves with
an atom, the absorbed energy causes one or
more electrons to shift positions within the
atom. When the electron goes back to its
original position, electromagnetic energy is
produced (Skoog, 2013). A pierces through a prism, it emits the colors
spectrophotometer is a device that precisely
measures electromagnetic energy at certain
wavelengths of light. It utilizes the intensity
of light and energy properties to determine
the maximum absorbance of the sample
(Phillips, 2024). Spectrophotometry is a
quantitative measurement method that
enables researchers to explore the optical
characteristics of materials throughout a
broad wavelength range, from ultraviolet to
visible and infrared. This method can be
used in determining the properties,
concentrations, materials, and even the
functional group present in the molecule
with the use of absorbance, transmittance,
and emittance (Qiu et al., 2010).
The human eye can perceive a part of the
electromagnetic spectrum called the visible
region or visible light where it has a
wavelength of 380 to 700 nm. As white light
of the rainbow that have its corresponding
wavelength. The principle of color is
generally about the absorption of selective
components of the wavelengths of the
visible light spectrum and emitting the
remaining wavelength that was not absorbed
(Harris, 2016).
 Figure 2: Color wheel wavelength (nm)
Figure 1: Structure of Bromocresol Green
As shown in figure 1, Bromocresol Green Methodology
(BCG) is composed of a methane group and Table 1: Materials and Instruments
three phenyl groups attached to a central Latex gloves 50-mL beaker
carbon atom. BCG is an odorless molecule Test tubes Cuvette
and when it is solid, it appears as yellowish 100-mL volumetric 10-mL Mohr pipet
purple in color and blue when it's diluted. It flask
belongs to a class of dyes called 3-mL volumetric UV-Vis
sulfonepthtaleins, a pH-sensitive indicator pipet spectrophotometer
that has a pH range of 3.8 - 5.4 that is useful 25-mL volumetric
in the medical field (NCBI, 2023). pipet
Table 2: Reagents
The objective of this study is to understand
0.001 M Distilled water
the theory and principles of
Bromocresol Green
spectrophotometry to determine the
(BCG)
wavelength of the maximum absorbance of
bromocresol green (BCG) using the UV-Vis
spectrophotometer. According to Djebbar et
A. Solution preparation for Standard
al., the maximum wavelength absorbance of
BCG solution
Bromocresol green is 616 nm. The
After the calculation, 30-mL of 0.01 M BCG
bromocresol green prepared in this study
stock solution was needed to dilute it to
was diluted, emitting a blue-green color as
3.0x10^-4 in a 100-mL volumetric flask. 25-
shown in Figure 2. The complementary of
mL and 5-mL volumetric pipet was used to
blue green is orange and has a wavelength of
transfer the required volume to produce the
590-630 nm. Thus, the possible wavelength
desired concentration.
that this experiment will get should fall in
the range of 590-630 nm wavelength as
B. Determination of Lamda max of BCG
shown in Figure 3. If the concentration is
standard solution
directly proportional to absorbance, then the
The standard solution was transferred to a
series of dilutions performed in this study
cuvette using a dropper and was loaded into
should emit a blue (standard BCG) to faint
the spectrophotometer. The visible range
blue (diluted BCG solution).
spectrum was set to 400-700 nm. The
absorbance was recorded for every 10 nm
and inserted into the Excel to create a chart.
a quantitative measurement method that
enables researchers to explore the optical
characteristics of materials throughout a
broad wavelength range, from ultraviolet to
visible and infrared.
indicates the absorption of orange and
emittance of the color blue. The value is also
close to the maximum wavelength
absorbance of the literature with some
potential errors during the study.

C. Relationship between absorbance and

concentration
Four (4) test tubes were prepared. Using a
Mohr pipet, 10-mL of stock solution was
transferred in the first test tube, 8-mL for the
second, 6-mL for the third, and 4-mL for the
fourth test tube. Distilled water is added
until the test tubes reach 10-mL in total. The
absorbance of each test tube was determined
using the spectrophotometer, data were
collected and recorded to create a calibration
graph.
Figure 4: Standard BCG solution
Results and Discussions
Table 3: Absorbance of different
concentrations

Ratio Absorbance Concentration

(10mL)
BCG: H2O
10:0 0.282 1.5x10-4
8:2 0.212 1.2x10-4
6:4 0.162 9.0x10-4
4:6 0.123 6.0x10-4
Figure 3: 𝜆max of Bromocresol Green
The four (4) test tubes were filled with a
(BCG)
ratio of the following: 10:0 mL, 8:2 ml, 6:4
mL, and 4:6 mL using a standard solution
Based on the figure 2, the complementary of
and diluting until it reaches 10-mL in total
blue is orange and has a wavelength range of
volume. In terms of qualitative results, the
590-630 nm. As shown in figure 4,
appearance of the test tubes shows the 1 st
bromocresol green emits a tint color of blue,
test tube having a blue appearance and as we
which means that it absorbs orange to emit
go towards the 4th test tube, the color slowly
the color blue. The literature states that
fades, turning into a faint blue. The
bromocresol green has a wavelength of 614
quantitative result shows the relationship
nm. According to the graph in Figure 3, the
between absorbance and concentration
result of the UV-Vis spectrophotometer
because as the concentration decreases, the
gave a maximum wavelength value of 610
absorbance value will also decrease as
nm and 0.281 for its absorbance. The
shown in Table 1. Therefore, the
experimental value falls under the range of
concentration is directly proportional to the
590-630 nm in the color wheel, which
absorbance value. This direct proportionality reasons for the experimental value to deviate
is the Beer-Lambert’s Law. a bit towards the value of one of the
literature’s values about bromocresol green.

Figure 5: Concentration and Absorbance

relationship

Conclusion
Spectrophotometry is a method that involves
the molecule’s capability to absorb light,
emitting the color that was not absorbed. It
can be measured by the intensity of light that
was absorbed by the molecule using a
spectrophotometer. This method helped this
study to determine the wavelength and
absorbance of the standard and series of
dilutions of bromocresol green (BCG). The
result obtained from this study after
determining the wavelength of standard
bromocresol green is 610 nm and has an
absorbance value of 0.281. Comparing to the
value obtained in one of the literatures, the
experimental value obtained from this study
was significantly close. The concentration of
the test tubes are as follows: #1: 1.5x10-4 M,
#2: 2.12x10-4 M, #3: 9.0x10-4 M, and #4:
4.60x10-4 M with an absorbance value of #1:
0.282, #2: 0.212, #3: 0.162, and #4: 0.123.
These values obtained after the series means
that as the concentration of the standard
BCG decreases, the absorbance value will
also decrease. This indicates the relationship
of concentration and absorbance to be
directly proportional with each other.
Personal errors such as personal
measurements, transfer of standard BCG to
the test tubes for dilution are possible
References
Fassi, S., Bousnoubra, I., Sehili, T., & Djebbar, K. (2012). Degradation of ’’ Bromocresol

Green’’ by direct UV photolysis, Acetone/UV and advanced oxidation processes (AOP’s)

in homogeneous solution (H 2 O 2 /UV, S 2 O 8 2-/UV).comparative study. J. Mater.

Environ. Sci, 3(4), 732–743.

https://www.jmaterenvironsci.com/Document/vol3/vol3_N4/74-JMES-215-2011-

Fassi.pdf
Harris, Daniel C. (2016). Quantitative Chemical Analysis Ninth Edition (9th). New
York: W.H.Freeman and Company.

HZDG. (2022). Hunterlab.com.

https://www.hunterlab.com/blog/what-is-spectrophotometer/#:~:text=A

%20spectrophotometer%20is%20a%20device

PubChem. (n.d.). Bromocresol green. Nih.gov. Retrieved February 3, 2023, from https://pub-

131chem.ncbi.nlm.nih.gov/compound/Bromocresol-green

Skoog, Douglas A., West, Donald M., Holler, F. James, Crouch, Stanley

R.. (2014). Fundamentals of Analytical Chemistry (Ed. 9th). Singapore: Cengage

Learning.

ultraviolet-visible uv-vis spectrophotometry: Topics by Science.gov. (n.d.). Www.science.gov.

Retrieved February 19, 2024, from https://www.science.gov/topicpages/u/ultraviolet-

visible+uv-vis+spectrophotometryFassi, S., Bousnoubra, I., Sehili, T., & Djebbar, K.

(2012). Degradation of ’’ Bromocresol Green’’ by direct UV photolysis, Acetone/UV and

advanced oxidation processes (AOP’s) in homogeneous solution (H 2 O 2 /UV, S 2 O 8

2-/UV).comparative study. J. Mater. Environ. Sci, 3(4), 732–743.

https://www.jmaterenvironsci.com/Document/vol3/vol3_N4/74-JMES-215-2011-

Fassi.pdf
Appendices
Calculations:

A. Solution preparation for standard BCG solution

M 1 V 1= M 2 V 2
1. M1 = 0.001 M
V1 = 100.0-mL
M2 = 3.0x10-4
V2 = ?
−4
(3.0 x 10 M )(100.0 mL)
V 1= =30.0 mL
0.001 M

B. Relationship between absorbance and concentration

1. Test tube #1 (10:0)

−4
(3.0 x 10 M )(10.0 mL )
M 1= =3.0 x 10−4 M
10.0 mL
2. Test tube #2 (8:2)
−4
(3.0 x 10 M )(8.0 mL) −4
M 1= =2.4 x 10 M
10.0 mL
3. Test tube #3 (6:4)
−4
(3.0 x 10 M )(6.0 mL)
M 1= =1.8 x 10− 4 M
10.0 mL
4. Test tube #4 (4:6)
−4
(3.0 x 10 M )(4.0 mL) −4
M 1= =1.2 x 10 M
10.0 mL
Table 4: 400-700 nm wavelength and their absorbances

Wavelength (nm) Absorbance

400 0.052
410 0.047
420 0.036
430 0.025
440 0.016
450 0.012
460 0.012
470 0.015
480 0.021
490 0.027
500 0.036
510 0.046
520 0.058
530 0.073
540 0.091
550 0.111
560 0.136
570 0.136
580 0.191
590 0.223
600 0.256
610 0.281
620 0.281
630 0.249
640 0.191
650 0.129
660 0.08
670 0.045
680 0.025
690 0.014
700 0.009