345

Indonesian Journal of Science & Technology 8(2) (2023) 345-362

Indonesian Journal of Science & Technology

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How to Calculate and Measure Solution Concentration

using UV-Vis Spectrum Analysis: Supporting Measurement
in the Chemical Decomposition, Photocatalysis,
Phytoremediation, and Adsorption Process
Asep Bayu Dani Nandiyanto1,*, Risti Ragadhita1, Muhammad Aziz2

1
Universitas Pendidikan Indonesia, Jl. Setiabudi No. 229, Bandung, Indonesia
2
The University of Tokyo, Tokyo, Japan
Correspondence: E-mail: [email protected]

ABSTRACT ARTICLE INFO

Article History:
UV-visible (UV-Vis) spectroscopy is a powerful instrument for Submitted/Received 20 Jan 2023
First Revised 10 Mar 2023
qualitative investigation and quantitative detection of Accepted 22 May 2023
pollutants in water. UV-Vis spectrophotometry is an First Available online 23 May 2023
analytical method using the concept of transmission of light Publication Date 01 Sep 2023

in UV and Visible wavenumber. Generally, compounds can be ____________________

Keyword:
identified using UV-Vis Spectrophotometry, based on the Absorbance,
concept of light absorption, specifically for compounds with Calibration curve,
a chromophore group and an auxochrome group. Although Concentration,
Photocatalyst,
the utilization of UV-Vis spectrum analysis has been well- Quantitative analysis,
documented, no information regarding detailed step-by-step Spectrophotometry,
measurement for examining detailed quantitative analysis, UV-Vis,
Wastewater treatment,
particularly in determining the concentration of an analyte in Wavelength.
an aqueous solution sample. Here, this study explores the
idea and application of UV-Vis technology in water quality
detection, including guidelines for determining the
concentration of the sample in an aqueous solution. To
support the analysis, we also added practical examples for
understanding concentration during the organic
decomposition. This paper is intended to be useful for
researchers and students in understanding UV-Vis
spectrophotometry when analyzing chemical composition
during chemical decomposition, photocatalysis,
phytoremediation, and adsorption analysis.

© 2023 Tim Pengembang Jurnal UPI

 Nandiyanto et al., How to Calculate and Measure Solution Concentration using UV-Vis … | 346

1. INTRODUCTION

In recent years, the degradation of various determining pollutant content in the

types of organic and inorganic pollutants in laboratory.
water using photocatalytic and adsorption However, these methods need some
methods have been studied extensively (Yu & instruments with large, complicated, and
Huang, 2023; Behera et al., 2021; Gautam et expensive apparatuses as well as a
al., 2020; Tufail et al., 2021; Hu et al., 2020). requirement of a high amount of reagents
Photocatalytic and adsorption methods have that sometimes create issues in the
been studied extensively due to their production of secondary pollution as a
relatively high activity, biological and byproduct. Further, the results are
chemical stability, low cost, simplicity, and somewhat not of a real-time process.
non-toxic (Behera et al., 2021; Sanakousar et Biological methods primarily include
al., 2022). enrichment analysis and biosensor
The determination and trace of organic technology; however, detection accuracy
and inorganic content in water are and sensitivity are significantly lower than in
interesting. For example, some inorganic other methods. Physical methods primarily
compounds in water are important and some include hyperspectral remote sensing
are toxic (Saravanan et al., 2021). Metals technology and molecular spectroscopy
such as zinc, manganese, copper, chromium, technology (Olaniran et al., 2013; Guo et al.,
and iron-cobalt are essential elements for 2020).
humans, animals, and plants. On the other Among the above methods,
hand, lead, cadmium, nickel, arsenic, and spectrophotometry is a well-recognized
mercury are toxic even at low levels method for identifying substances. It
(Mehrandish et al., 2019). Then, the organic conducts quantitative determinations using
content causes a lack of dissolved oxygen the emission or absorption spectra of
which can cause death in aquatic biota due to substances. It has been widely used in the
lack of oxygen (Mahaffey et al., 2020). field of rapid water quality determination in
Therefore, it is necessary to estimate the recent years.
organic and inorganic content in the solution, Recently, due to the advantages of high
particularly in the aqueous sample. precision, high detection efficiency, non-
Many analytical techniques exist in the destructive sampling, environmental
literature for being utilized in the protection, low cost, and portability,
measurement and traces of heavy metals ultraviolet-visible (UV-Vis) technology has
and organic compounds in mixed solutions, been developed into an advantage and an
especially in the aqueous solution. The effective tool for detecting pollutants in
classification of methods for the aqueous environments (Guo et al., 2020;
determination of water quality is shown in Pratiwi & Nandiyanto, 2021). Some
Figure 1. Chemical, biological, and physical researchers have attempted to summarize
methods are currently used to determine the application of UV-VIS spectroscopy for
water quality parameters. Chemical oxygen determining the chemical structure of
demand (COD), heavy metal content, nitrate compounds previously. Some researchers
nitrogen (NO3-N), dissolved organic carbon also deepened UV-Vis spectroscopy for
(DOC), and turbidity are the most important determining the chemical structure of
water quality parameters. Titration analysis compounds. However, discussion in the field
and electrochemical analysis are the two of water quality detection, research objects,
most common chemical methods for and research methods is still limited.

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Figure 1. Methods for Determining Water Quality (adopted from Guo et al., 2020).
Therefore, this study introduces the (Albert et al., 2012). This absorbance value is
theoretical basis for determining various used in determining the amount of
parameters of water quality by UV-Vis concentration (for quantitative analysis) and
spectroscopy and outlines the complete type of component contained in a sample
spectral data analysis process, including data (for qualitative analysis) (Bardik et al., 2020).
pre-processing, characteristics of wavelength Light absorbed by a substance is different
extraction, and absorbance formation. All from the light captured by the human eye.
explanations were introduced step by step to Visible light (or light seen in our daily life) is
make clear and easily understand called complementary colors. For example, a
researchers and students for understanding substance will be orange if it absorbs blue
solution concentration in aqueous solution, from the visible light spectrum and a
especially when analyzing chemical substance will be black if it absorbs all colors
composition during chemical decomposition, contained in the visible light spectrum. For
photocatalysis, phytoremediation, and more details, the absorbed colors and
adsorption analysis. complementary colors are shown in Table 1
(Akash et al., 2020).
2. BASIC PRINCIPLES
Based on the Lambert-Beer law, the
2.1. Theoretical of UV-VIS
working principle of the UV-Vis
Spectrophotometer in Water
spectrophotometer is that polychromatic
Treatment
light from a light source will be converted
UV-Vis spectrophotometer is a into monochromatic light using a
spectrophotometric technique in the area of monochromator. The light is partially
ultraviolet and visible light. This tool is used absorbed by the cell in the sample and some
to measure the absorption of light ultra- light will be passed through the photocell
violet and visible light in a sample. This type based on a certain wavelength. The amount
of spectrophotometer is more widely used in of light that passes through this will be
analyzing a material quantitatively by counted by the detector. The absorbance
measuring the light absorbance value of a value of the light that is passed will be
sample based on a specified wavelength proportional to the concentration of the
solution in the cuvette (Guo et al., 2020).

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Table 1. The Absorbed Colors and Complementary Colors.

Wavelength (nm) Absorbed colors Complementary colors (visible colors)
390-450 Violet Yellow
450-495 Blue Orange
495-570 Green Red
570-590 Yellow Violet
590-620 Orange Blue
620-750 Red Green

In detecting the concentration of analyte which the transmission beam of light will
in the aqueous solution, UV-Vis spectroscopy increase. Indeed, the absorption also
is based on the fact that at certain increases. The Lambert-Beer Law must
wavelengths, pollutant molecules in water comply with the requirements in its use. The
can absorb UV-Vis light. Light at certain terms of the Lambert-Beer Law, namely the
wavelengths is absorbed due to the amount of concentration used is not extreme
movement of electrons from the ground (the sample used is not highly concentrated),
state to the excited state, which reduces the the sample to be analyzed must not
amount of light transmitted. Pollutants dissociate, associate, or react with solvents
primarily absorb light in the UV-Vis range. As that will create other products. Or, the color
a result, we can detect the concentration of formed must be stable. The absorption of
pollutants in water using the Lambert-Beer light by the specified substance and the light
law as a theoretical foundation. Organic measured is monochromatic light (Akash et
matter and turbidity are effectively adsorbed al, 2020). In short, the Lambert-Beer Law,
in the 380-750 nm wavelength range (Guo et namely the concentration of the solution
al., 2020). The range of absorption spectra being analyzed is directly proportional to the
and the substance characteristics of the amount of light (absorbance) absorbed by
various substances are shown in Table 2. the substance contained in the solution. The
wavelength used in this tool ranges from 200
2.2. Lambert-Beer Law
to 700 nm. The color absorbed by a
According to Lambert, absorption is compound or element is complementary. A
directly proportional to the thickness of the schematic diagram of the Lambert-Beer law
irradiated cell, while according to Beer, is shown in Figure 2.
absorption is directly proportional to Based on Figure 2, the outgoing light (I)
concentration. These two statements are put correlates to solution turbidity and depends
together in the Lambert-Beer Law, namely, on the intensity of the incident light (I0). K is
absorption is directly proportional to the the molar absorption coefficient related to
concentration and thickness of the cell. This the nature of the absorbing substance and
is because if the cell increases, the the wavelength λ of the incident light. a is the
absorption will increase. If the concentration concentration of the absorbing substance in
increases, the number of molecules through mol/L. l is the thickness of the absorbing layer
in cm.
Table 2. The Range of Absorption Spectra and the Substance Characteristics of the Various
Substances.
Wavelength Range (nm) Material Characteristics
200-220 Nitrate, nitrite
220-250 Conjugated diene, unsaturated aldehyde, unsaturated ketone
250-750 Organic matter and turbidity

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Figure 2. Schematic Diagram of the Lambert-Beer Law.

Absorbance is the amount of light or 𝐴 = 𝑎 × 𝑙 × 𝑐 𝑜𝑟 𝐴 = ɛ × 𝑙 × 𝑐 (4)
energy absorbed by the particles in the By rearranging Eq. (4), the Lambert-Beer
solution (the relative amount of light Law makes it possible to determine the
absorbed by the sample). The transmittance concentration of a sample from a measured
is the portion of the light that is transmitted absorbance value. Molar absorptivity
(the relative amount of light that passes constant (ε) and path length (l) are known.
through the sample). The relationship The concentration of c can be calculated
between absorbance and transmittance is from the absorbance of A using Eq. (5).
inversely proportional. The higher 𝐴
transmittance results in the lower the 𝑐 = ɛ×𝑙 (5)
absorbance obtained, and vice versa. This is where 𝐴 is absorbance, 𝑙 is the thickness of
found in the Lambert-Beer Law, that is, if light the cuvette is also generally 1 cm, 𝑐 is the
passes through a solution without concentration of the solution measured.  is
experiencing absorption, the absorption will the molar absorptivity constant (if the
be zero. If all light is absorbed, the concentration of the solution measured is in
transmittance is zero and the absorbance is molar terms) or the molar absorptivity
infinity. The absorbed light is measured as constant (if the concentration of the solution
absorbance (A), while the scattered light is measured is in mg/L terms).
measured as transmittance (T). All light The absorbance must be within the linear
phenomena are expressed by Lambert-Beer's range of the instrument for optimal
law (Aljamali, 2015; Akash et al., 2020). measurement results following the Lambert-
Based on Lambert-Beer's law, the formula Beer's Law. The optimal measurement range
used to calculate the amount of light (i.e. the measurement range over the
scattered is shown in Eq. (1) and Eq. (2). absorbance) is directly proportional to the
𝐼
𝑇 = 𝐼𝑡 (1) concentration. It is given as 0.3 < A < 2.5
0
(Mousa et al., 2017).
Or
𝐼
Therefore, it is advisable to avoid very high
%𝑇 = 𝐼𝑡 × 100% (2) absorbance values (A > 2.5) as well as very
𝑜

and absorbance is expressed by Eq. (3). low absorbance values (A < 0.3), which may
𝐼 result in a non-linear behavior of the
𝐴 = − log 𝑇 = −𝑙𝑜𝑔 𝐼𝑡 (3) calibration line. This is illustrated in Figure 3,
𝑜

The formula derived from the Beer's Law can where the measured values of above A = 2.5
be written by Eq. (4). and below A = 0.3 (red dotted line) deviate
from the theoretical calibration line (green):
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Figure 3. Non-Linearity: The Red Measured Values Outside the Linear Range Deviate from
the Green Theoretical Calibration Line.
2.3. Measurement Principle • The intensity of the transmitted light
at different wavelengths is then
A UV/VIS spectrophotometer compares
measured by a detector positioned
the intensity of light passing through a
after the cuvette with the solvent and
sample solution in a cuvette to the intensity
recorded.
of light before it passes through the sample.
(ii) Sample determination. There are steps
A light source, a sample holder, a dispersive
for sample determination:
device (e.g., a monochromator), and a
• A sample dissolved in the solvent is
suitable detector are the main components
added to the cuvette.
of a V/VIS spectrophotometer (Akash et al.,
• A light beam emitted by the light
2020). Measurement principle in UV/VIS
source passes through the cuvette
spectroscopy shown in Figure 4. The working
with the sample.
principle of a spectrophotometer is based on
blank measurement and sample • When passing through the cuvette,
determination. The following steps are: the light is partially absorbed by the
(i) Blank (a measure of the intensity of light sample molecules in the solution.
transmitted through the solvent). Blank • The transmitted light is then
measurement is needed for the sample measured by the detector.
concentration determination. There are • The light intensity change at different
steps for blank measurement: wavelengths is calculated by dividing
• The solvent (e.g. water or alcohol) is the transmitted intensity of the
added into a suitable, transparent, sample solution by the
and not absorbing container (a corresponding values of the blank.
cuvette). This ratio is finally stored by a
• A light beam emitted by the light recorder.
source passes through the cuvette
with the solvent.

Figure 4. Measurement Principle in UV/VIS Spectroscopy.

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2.4. Important Aspects to be Considered A standard solution is any chemical

During the Measurements solution that has a precisely known
concentration. Similarly, a solution with
The aspects that need to be considered in
known concentration needs to be
spectrophotometer measurements, namely
standardized. To prepare a standard
the sample used must be colored. If the
solution, we dissolve a known mass of solute
solution to be analyzed is colorless, the
and then dilute it to the correct volume of
sample must first be colored. It is intended
solution. Standard solution concentration is
that the sample can be analyzed based on its
usually expressed in terms of molarity (M) or
color-forming substances (Yang et al., 2017).
moles per liter (mol/L). Usually, the solution
The wavelength used also needs to be
is diluted with distilled water or another
considered. The level of error in the
solvent (such as methanol, ethanol, etc.) to
measurement will be less if the maximum
the limit and homogenized until a standard
wavelength is used. This is because, at the
solution of 100 ppm is obtained. Not all
maximum wavelength, the level of sensitivity
substances are suitable solutes for standard
of a sample will be maximum. In addition, by
solutions. The reagent must be stable, pure,
using the maximum wavelength, the
and preferably of high molecular weight.
absorbance curve obtained will comply with
(ii) Step 2: Determination of the maximum
the Lambert-Beer law. In addition to these
wavelength
two aspects, the calibration of the
Before calculating the sample
wavelength and absorbance needs attention.
concentration using a UV-Vis
A spectrophotometer is used to measure the
spectrophotometer, we determine the
intensity of light. Each sample used has a
maximum wavelength with the aim to be
different light absorption depending on the
able to provide maximum sensitivity of
compound formed. Therefore, it is necessary
samples containing analytes. Detailed
to calibrate to obtain more accurate results
determination of the maximum wavelength
(Guo et al., 2020).
is carried out as follows:
2.5. Step-by-Step Quantitative Analysis • The spectro instrument is set to quantity
using UV-VIS using Standard Curves mode and the wavelength is set.
• The wavelength is set and adjusted to the
One of the most common quantitative
color of the complementary adsorbate
methods of analysis in water treatment is the
solution to be measured.
determination of an analyte's concentration
• Before measuring the absorbance of the
based on the absorption of ultraviolet or
standard adsorbate solution, the blank
visible radiation that accumulates in water.
solution is measured first at a
One reason is that many organic and
predetermined wavelength. The blank
inorganic compounds have a strong
solution is a solution that does not
electromagnetic spectrum absorption band
contain analytes or a solution without a
in the UV/visible region (such as organic and
sample. It is usually only solute or 0% of
inorganic compounds). Furthermore,
the analyte.
analytes that do not absorb or weakly absorb
• Then, the absorption of approximately 3
UV/Vis radiation can frequently be
mL of standard solution was measured
chemically combined with species that do
using a UV-Vis spectrophotometer at
absorb ultraviolet and visible light.
maximum wavelength. Usually, the
Quantitative analysis steps such as
cuvette contains about 3 mL.
determining the concentration of analytes in
• After that, the absorbance value of each
a sample are generally described as follows:
measured wavelength is recorded and
(i) Step 1: Preparation of standard solutions
tabulated (see Table 3).
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Table 3. Example of Tabulated Wavelength vs Absorbance.

Wavelength (nm) Absorbance
200 0.125
250 0.273
300 0.294
And so on And so on

• Finally, the determination of the diluted solution, and M2 is the

maximum wavelength is taken from the concentration of the solution resulting
measured wavelength to produce from the dilution. Detailed calculations
maximum absorbance. for the manufacture of serial
(iii) Step 3: Determination of the calibration concentrations of standard solutions are
curve as follows:
Making a calibration curve is used to find 𝑉1 𝑀1 = 𝑉2 𝑀2
a linear regression equation. Thus, it can be 𝑉1 100 𝑝𝑝𝑚 = 100 × 2
used to find sample levels of measured 𝑉1 = 2 𝑚𝐿
absorbance. To create a detailed calibration Thus, to make a 2-ppm standard solution
curve, the steps are as follows: series with a volume of 100 mL, 100-ppm
• Making a calibration curve is done by stock solution is taken as much as 2 mL.
preparing a series of standard solutions • A series of standard solutions with a
with a certain concentration. As certain concentration has been prepared.
previously explained, the calibration
Then, the absorbance is measured at the
curve aims to find a linear regression
wavelength determined in the previous
equation. This linear regression equation
step in determining the maximum
is determined from the relationship wavelength. Finally, the absorbance
between the analyte concentration series values of each series of standard
and the absorbance of the analyte. A
solutions are recorded.
series of standard solutions with a certain
• After the absorbance values of a series of
concentration is prepared by the dilution
standard solutions are obtained, the
process of standard solutions. For
absorbance values are plotted to become
example, working standards are made by
a curve of the relationship between the X
making a series of six solutions. Each
and Y-axis curve. The series of analyte
solution has a concentration of 2, 3, 4, 5,
levels is plotted on the Y-axis and the
and 6 ppm. Each set of standard solutions
absorbance of the analyte is plotted on
is made by taking as much as 2; 3; 4; 5;
the X-axis. Figure 5 is a calibration curve
and 6 mL of standard solution
determined by relating the series of
respectively, then transferred to a 100-
analyte levels to the absorbance of the
mL volumetric flask and dissolved with
analyte. Figure 5 has an intercept value
distilled water to the limit and
(a) = 0.062 and a slope value (b) = 0.017
homogenized. To obtain a series of
with a correlation value (r) = 0.998.
standard solutions with concentrations of
Therefore, the linear equation is y =
2, 3, 4, 5, and 6 ppm from a 100-ppm
0.062x – 0.017. The resulting absorbance
standard solution, it is calculated using
data is quite good because all series
the dilution formula as in Eq. (6).
contents from the smallest to the largest
𝑉1 𝑀1 = 𝑉2 𝑀2 (6) values have an absorbance value of 0.2 –
where V1 is the volume of stock solution 0.8, while the correlation value obtained
taken, M1 is the concentration of the is 0.998. This curve is very good because
diluted solution, V2 is the volume of the a good correlation value is close to 1.
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Figure 5. Example of The Calibration Curve.

(iv) Step 4: Determination of Sample previously obtained from the calibration
Concentration using Calibration Curve curve determination step (step 3). The
To determine the concentration of the sample absorbance data is then
analyte in the sample, the details of the steps substituted into the linear equation (y =
are as follows: mx + c) which has been obtained from the
• Before measuring the absorbance of the calibration curve based on the
sample, the blank solution is measured absorbance of the standard solution
first at a predetermined wavelength. The series. Assuming the sample absorbance
blank solution is a solution that does not data is the y variable, the x value is
contain analytes or a solution without a obtained as the concentration value.
sample. For example, in the case of Based on the linear equation obtained
analyzing the concentration of curcumin from the calibration curve (step 3), the
in the aqueous solution, because detailed sample concentration is
curcumin is dissolved in pure water, the calculated as follows:
blank solution is pure water. 𝑦 = 0.062𝑥 − 0.017
• The sample (or example a water sample 0.269 = 0.062𝑥 − 0.017
containing organic or inorganic 0.269 + 0.017
contaminants) is put into a cuvette and 𝑥= = 4.61 𝑝𝑝𝑚
0.062
its absorbance is measured at the
Thus, the analyte concentration in the
maximum wavelength determined in the
sample is 4.61 ppm.
maximum wavelength determination
(v) Step 5: Determination of Sample
step (step 2).
Concentration using The Initial
• After the absorbance value is obtained,
Concentration Peak
the absorbance value is recorded. The
This method can be used as a shortcut for
absorbance value will correspond to the
analyzing the concentration without
concentration of the sample according to
additional calibration process. However, we
the Lambert-Beer law. For example,
should understand the initial concentration
measuring a sample yields an absorbance
of solution.
value of 0.269.
To determine the concentration of the
• Next, the calculation of the concentration
analyte in the sample using this method, the
of the analyte in the sample is carried out details of the steps are as follows:
using the linear equation that was

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• Before measuring the absorbance of the concentration of pollutant samples

sample, the blank solution is measured accumulated in water in water treatment
first at a predetermined wavelength. The applications. A detailed step-by-step
blank solution is a solution that does not approach for quantitative analysis in
contain analytes or a solution without a determining the concentration of a sample of
sample. For example, in the case of a pollutant in the aqueous solution is
analyzing the concentration of curcumin described in this study.
in the aqueous solution, because
4. RESULTS AND DISCUSSION
curcumin is dissolved in pure water, the
4.1. Experimental Results
blank solution is pure water.
• The initial sample (or example a water One example of an organic dye used as a
sample containing organic or inorganic model for a pollutant in the case raised in this
contaminants) is put into a cuvette and its study is curcumin. In short, the water
absorbance is measured at the maximum treatment process is carried out by reducing
wavelength determined in the maximum the concentration of dyes as a contaminant
wavelength determination step (step 2). derived from curcumin which accumulates in
The concentration using this initial water through a series of water treatment
concentration is a mandatory. The curve processes (e.g., photocatalyst,
from initial concentration will be used as phytoremediation, or adsorption). Also, in
an initial standard for the next this study, water treatment to reduce or
calculation. remove pollutants such as organic dyes are
• Other samples (or example a water demonstrated.
sample containing organic or inorganic The success of the water treatment
contaminants) are put into one by one process is determined by how many
into a cuvette and their absorbance is pollutants accumulate in the water that can
measured at the maximum wavelength be removed or reduced. Thus, the initial
determined in the maximum wavelength (before treatment) and final (after
determination step (step 2). treatment) concentrations of pollutant
• After the absorbance value is obtained, samples are compared to evaluate the
the absorbance value is recorded. The success of the water treatment process. The
absorbance value will correspond to the water treatment process is said to be
concentration of the sample according to successful if the final pollutant sample
the Lambert-Beer law. For example, concentration is lower than the initial
measuring a sample yields an absorbance pollutant sample concentration. Therefore,
value of 0.269. the determination of the concentration of
• Next, the calculation of the concentration pollutant samples that exist in the sample
of the analyte in the sample is carried out was carried out before and after the
by comparing to the initial concentration. treatment was carried out.
The calculation using Eq. (7). However, here, to analyze the success of
𝐶𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝑋 = the process, the determination of the sample
𝐴𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒 𝑋
𝑥 𝐶𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝐼𝑛𝑖𝑡𝑖𝑎𝑙 (7) concentration is not only carried out before
𝐴𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒 𝐼𝑛𝑖𝑡𝑖𝑎𝑙
and after the treatment but the
3. METHOD determination of the sample concentration is
carried out at each time interval.
To understand how to carry out Determination of the concentration of a
quantitative analysis with UV-VIS sample of a solvent in an aqueous solution is
instruments, this study is equipped with based on the Lambert-Beer law of solids.
examples of how to determine the According to the Lambert-Beer law, the

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amount of spectral absorption has a strong solution was a standard curcumin solution
correlation with the concentration of water with a concentration of 100 ppm.
quality parameters. Thus, high-precision (ii) Step 2: Determination of the maximum
prediction models between water quality wavelength
spectral data and water quality parameters The steps for determining the maximum
can be created (Li et al., 2018). wavelength are as follows:
• The spectro instrument is set to quantity
4.2. Calculation Procedures (Step-by-Step
mode and the wavelength is set.
Process): Example 1
• The wavelength is set and adjusted to the
In determining the concentration of the color of the complementary adsorbate
sample by UV-Vis spectroscopy, the solution to be measured. Based on Table
experiment "determination of the 1, because the curcumin solution has a
concentration of curcumin accumulated in yellow-to-orange complementary color,
water after the water treatment process (i.e., the color that will be absorbed is violet to
photocatalysis or adsorption processes)" is blue. Therefore, the maximum
exemplified step by step. wavelength is set at 390-495 nm.
The steps for determining the • Then, standard adsorbate solutions with
concentration of curcumin dye solution with concentrations of 100-ppm were then
UV-Vis are divided into 2 stages, namely the measured for absorbance using a UV-Vis
pre-treatment stage and the post-treatment spectrophotometer at λ = 390-495 nm.
stage. In the pre-treatment stage, three steps The principle of determining the
must be carried out, namely preparing a maximum wavelength is by scanning the
standard solution (step 1), determining the absorbance value in the desired
wavelength (step 2), and preparing a wavelength range. For example, here, the
calibration curve (step 3). Meanwhile, the desired wavelength range is 390-495 nm.
post-treatment stage only includes one step, Thus, the maximum wavelength scanning
namely determining the sample process is carried out in the desired
concentration (step 4). wavelength range, namely 390-495 nm.
To determine the concentration of organic • After that, the absorbance value of each
dyes (such as curcumin) as pollutants interval wavelength is measured (starting
accumulated in water using a UV-Vis from 390-495), recorded, and tabulated
instrument, the steps are as follows: as exemplified in Table 3.
(i) Step 1: Preparation of standard solution • After scanning the wavelength in the
To prepare a standard solution, dissolve a desired range, several absorption peaks
known mass of solute diluted and will appear as shown in Figure 6. Finally,
homogenized to the correct volume of the determination of the maximum
solution. To prepare a 100-ppm standard wavelength is taken from the measured
solution, as a model organic dye, curcumin wavelength to produce maximum
powder was used. A 100-ppm curcumin absorbance. Here, the selected maximum
standard solution was prepared by dissolving wavelength of the curcumin solution is
and homogenizing 5 grams of curcumin 395 nm due to has higher absorbance
powder with 600 mL of purified water. After value. Because the maximum wavelength
being homogeneous, the 100-ppm curcumin is known, the absorbance measurement
solution was filtered through a vacuum filter for the standard curve (in step 3) and
to separate the insoluble curcumin residue. concentration (in step 4) is measured at
The filtrate resulting from the curcumin this maximum wavelength, which is 390
nm.

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Figure 6. Some of The Absorption Peaks Result from Scanning the Wavelengths.
(vi) Step 3: Determination of the calibration Thus, to obtain a series of standard
curve solutions with a concentration of 20 ppm
Before determining the calibration curve, with a volume of 10 mL, the standard
there are several steps: 100-ppm curcumin solution that must be
• The first step that must be taken is to taken is 2 mL. Then, the same method
prepare a series of standard solutions was adopted for the preparation of other
with certain concentration variations. A series of standard solutions with
series of standard solutions are prepared concentrations of 30, 40, 60, and 80 ppm.
by determining the series of standard The dilution method for preparing a
solutions to be made from 100-ppm standard adsorbate solution with a
curcumin standard solution. For example, concentration of 20 ppm is also
the concentration series of standard analogous to preparing a standard
solutions to be made are 20, 30, 40, 60, adsorbate solution with a concentration
and 80 ppm. After that, a series of of 30, 40, 60, and 80 ppm by taking 3.0;
standard solutions with concentrations of 4.0; 0.6; and 0.8 mL of the 100-ppm
20, 30, 40, 60, and 80 ppm were prepared curcumin standard solution, respectively.
by diluting 100 ppm curcumin standard • After a series of standard solutions were
solution. The dilution formula follows Eq. prepared, the next step was to measure
(1). An example of detailed calculations the absorbance with the UV-VIS
for making a standard solution with a instrument at the maximum wavelength.
concentration of 20 ppm with a volume The absorbance results are then recorded
of 10 mL from a standard solution of 100 and tabulated for each value of the
ppm with dilution is as follows: concentration of the solution.
𝑉1 𝑀1 = 𝑉2 𝑀2 Absorbance data from a series of
𝑉1 100 𝑝𝑝𝑚 = 10 × 20 variations in the concentration of
𝑉1 = 2 𝑚𝐿

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standard curcumin solutions are listed in initial concentration of the curcumin solution
Table 4. accumulated in water, the absorbance was
• The concentration and absorbance data measured before the water treatment
from a series of standard solutions were process (i.e., photocatalysis or adsorption
then converted into a calibration curve by processes). Meanwhile, to determine the
plotting the X-axis as concentration and final concentration of curcumin solution
Y-axis as absorbance. The standard curve accumulated in water, the absorbance was
of the solution obtained is shown in measured after the water treatment process
Figure 8. From Figure 8, the linear (i.e., photocatalysis or adsorption processes)
equation obtained is y = 0.0114x + 0.0189 was carried out. The hope is that through the
with an intercept value (a) = 0.0114 and a water treatment process, the dye or
slope value (b) = 0.0189 with a correlation pollutant compounds can be reduced or
value (R2) = 0.9794. removed from the water. Therefore, to prove
(iii) Step 4: Determination of Sample that the water treatment process is
Concentration successful, the initial and final concentrations
After steps 1-3 (pre-treatment stage) have of the sample solution must be determined
been carried out, the next step is to and compared. If the final concentration of
determine the concentration of the curcumin the sample is less than the initial
solution sample (pra-treatment stage). concentration of the sample, then the water
Determination of the concentration of treatment process is successful because
curcumin solution samples can be done to based on determining the concentration of
determine the initial and final concentrations the sample, the pollutants accumulated in
of curcumin solution samples that the water can be reduced or eliminated
accumulate in the water. To determine the (Nandiyanto et al., 2020).
Table 4. Absorbance and Concentration Data of Series Curcumin Standard Solution.
Concentration (ppm) Absorbance
20 0.210
30 0.348
40 0.538
60 0.707
80 0.903

Figure 8. Calibration Curve of a Series of Standard Curcumin Solutions from Photocatalyst

Applications.

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To measure the concentration of the 𝑦 = 0.0114𝑥 + 0.0189

sample, both initial and final concentrations, 0.761 = 0.0114𝑥 + 0.0189
the blank solution is first measured at a 0.761 − 0.0189
predetermined wavelength. After that, the 𝑥= = 65.096 𝑝𝑝𝑚
0.0114
absorbance of the samples before and after Thus, the final concentration of the
the water treatment was measured one by sample is 65.096 ppm. Examples of the
one by inserting the sample solution into the calculation results presented in Table 5.
cuvette and then measuring it with a UV-Vis (vii) Step 5: Determination of Sample
instrument. According to the Lambert-Beer Concentration using The Initial
law, the absorbance value will correspond to Concentration Peak
the concentration of the sample. The This method can be used as a shortcut for
absorbance measurement results for the analyzing the concentration without
initial and final solutions obtained values of additional calibration process. The initial
0.150 and 0.051 respectively. Furthermore, sample (or example a water sample
the calculation of the concentration of the containing organic or inorganic
analyte in the sample is carried out by contaminants) is put into a cuvette and its
substituting the absorbance value in the absorbance is measured at the maximum
linear equation previously obtained from wavelength determined in the maximum
step 3. Here, the absorbance data of the wavelength determination step (step 2). The
sample after the treatment process at each concentration using this initial concentration
time interval is exemplified and presented in is a mandatory. The curve from initial
Table 5. The detailed determination of the concentration will be used as an initial
concentration is exemplified for the standard for the next calculation.
calculation of the initial and final After the absorbance values are obtained,
concentrations of the sample solution by the absorbances will be compared to the
substituting the absorbance value into the initial concentration. For example, when we
linear equation which has been obtained understand the initial concentration is 78
from the calibration curve which can be ppm (absorbance = 0.912), measuring a
calculated as follows: sample with an absorbance value of 0.877
• Calculation of the determination of the can result concentration of 75 ppm. This
initial concentration value can be obtained by dividing 0.877 with
𝑦 = 0.0114𝑥 + 0.0189 0.912 and multiplying with 78 ppm. Detailed
0.912 = 0.0114𝑥 + 0.0189 calculation results are shown in Table 5.
0.912 − 0.0189 Detailed calculation is in the following:
𝑥= = 78.342 𝑝𝑝𝑚
0.0114 𝐶𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝑋
Thus, the initial concentration of the 𝐴𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒 𝑋
= 𝑥 𝐶𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝐼𝑛𝑖𝑡𝑖𝑎𝑙
sample is 78.342 ppm. 𝐴𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒 𝐼𝑛𝑖𝑡𝑖𝑎𝑙
0.877
• Calculation of the determination of the 𝐶𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝑋 = 𝑥 78 𝑝𝑝𝑚 = 75 𝑝𝑝𝑚
0.912
final concentration
Table 5. Example 1 of Absorbance Data of Pollutant Samples After Treatment at Each Time
Interval.
Time Concentration based Concentration based on Initial
Absorbance
(minutes) on Calibration (ppm) Concentration (ppm)
0 0.912 78.342 78.000
30 0.877 75.272 75.007
60 0.779 66.675 66.625
90 0.766 65.535 65.513
120 0.761 65.096 65.086

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4.3. Calculation Procedures (Step-by-Step time interval, respectively as evidenced by

Process): Example 2 Figures 9(a) and (b), corresponding to UV Vis
spectra and photograph image for the
Determination of the concentration for
decolorization, respectively. Using the above
each time interval is also calculated in the
procedures, we can convert the absorbance
same way as in the calculation of determining
into concentration in Table 6.
the concentration of the initial (before the
In this case, we cannot do regression. But
treatment, 0 minutes) and final (after the
we understand the initial concentration (at
process, 120 minutes) samples. Based on this
time of 0 min) is 100 ppm. Thus, we can
step, the initial and final concentrations of
convert the concentration at various times
the dye sample solution accumulated in
from absorbance. For example, when the
water have been identified. The results
reaction is 12 minutes and the absorbance is
showed that the final concentration of the
0.7, the concentration can be obtained by
sample solution decreased after the water
dividing 0.7 with 0.8 and multiplying with 100
treatment process. Based on the results
ppm (see equation (7)). The result will be 88
described, the success of the process is
ppm. Detailed calculation is in the following:
characterized by a reduced concentration of 𝐶𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝑋
dye contaminants accumulated in the water 𝐴𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒 𝑋
after the treatment process which is in line = 𝑥 𝐶𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝐼𝑛𝑖𝑡𝑖𝑎𝑙
𝐴𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒 𝐼𝑛𝑖𝑡𝑖𝑎𝑙
with the decrease in the absorbance value 0.7
𝐶𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝑋 = 𝑥 100 𝑝𝑝𝑚 = 88 𝑝𝑝𝑚
and color intensity of the solution at each 0.8

Figure 9. Evidence of the success of the curcumin solution process shown by the decrease in
the absorbance value (a) and the physical appearance of the intensity of organic dyes in
aqueous solution (b) (adopted from Sukmafitri et al., 2019).
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Table 6. Example 2 of absorbance data of samples

Concentration Results Based
Time (Minutes) Absorbance on Initial Concentration
(ppm)
0 0.79 100
12 0.70 88
48 0.55 69
84 0.29 36
120 0.05 6

5. CONCLUSION when analyzing chemical composition during

chemical decomposition, photocatalysis,
This paper shows the steps for phytoremediation, and adsorption analysis.
quantitative analysis using UV-VIS
6. AUTHORS’ NOTE
instruments, in particular the steps for
determining sample concentrations using
The authors declare that there is no
UV-VIS instruments are explained in detail.
conflict of interest regarding the publication
With this research, we believe that this paper
of this article. Authors confirmed that the
can be used as a basis for understanding the
paper was free of plagiarism.
determination of sample concentration in
aqueous solutions using UV-VIS, especially

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