Development of Fourier Transform Infrared Spectrophotometric

Nerdy Nerdy, et al.

Articles

MOLEKUL

eISSN: 2503-0310

https://doi.org/10.20884/1.jm.2025.20.2.8635

Development of Fourier Transform Infrared Spectrophotometric Method for Quantification of
Simvastatin, Atorvastatin, and Rosuvastatin in Marketed Tablet Preparations
Nerdy Nerdy1, Nilsya Febrika Zebua1, Syarifah Nadia1, Effendy De Lux Putra2, Tedy Kurniawan Bakri3
Department of Pharmacy, Faculty of Pharmacy and Health, Universitas Tjut Nyak Dhien, Sei Sikambing,
Medan Helvetia, Medan, Sumatera Utara, Indonesia, 20123
2
Department of Pharmacy, Faculty of Pharmacy, Universitas Sumatera Utara, Padang Bulan, Medan Baru,
Medan, Sumatera Utara, Indonesia, 20155
3
Department of Pharmacy, Faculty of Mathematics and Natural Sciences, Universitas Syiah Kuala, Kopelma
Darussalam, Syiah Kuala, Banda Aceh, Aceh, Indonesia, 23111
1

*Corresponding author email : nerdy@utnd.ac.id
Received June 09, 2023; Accepted June 19, 2025; Available online July 20, 2025
ABSTRACT. Marketed pharmaceutical dosage forms must be checked to ensure that the drug in distribution meets the quality
requirements. The Fourier transform infrared spectrophotometric method will provide new innovations which are low cost,
environmental friendly, simple, and rapid for analysis. The aim of this research was to develop a single analytical method
that can be used to determine levels of three different statin derivatives (simvastatin, atorvastatin, and rosuvastatin) in tablet
preparations. The method used in this study was Fourier transform infrared spectrophotometric, a very simple sample
preparation stage by solvent extraction with minimal solvent extraction without potassium bromide and without pellet press.
The specific wave numbers of simvastatin, atorvastatin, and rosuvastatin were determined between 4000 cm –1 to 650 cm–1.
The Fourier transform infrared spectrophotometric method had been successfully developed and successfully applied to the
determination of levels of simvastatin, atorvastatin, and rosuvastatin in marketed tablet preparations containing single active
pharmaceutical ingredient. The specific wave numbers of simvastatin, atorvastatin, and rosuvastatin obtained was 1712.7
cm–1; 1660.5 cm–1; and 965.4 cm–1, respectively. The results obtained in the content determination based on the peak height
and peak area fulfill the requirements for active pharmaceutical ingredient assay ranging from 90.00% to 110.00%. The
developed method has also been validated for accuracy, precision, specificity, linearity, range, limit of detection, and limit
of quantitation. Thus, this method can be a valid alternative method, cheap, green, easy, and fast in determination of the
levels of simvastatin, atorvastatin, and rosuvastatin in tablet preparations.
Keywords : atorvastatin, FTIR, rosuvastatin, simvastatin, tablet

INTRODUCTION
Drugs for the treatment of dyslipidemia that are
widely available commercially in pharmaceutical
tablet preparations are statin derivatives, likely
simvastatin, atorvastatin, and rosuvastatin (Sadowska
et al., 2024). Statin derivatives are the main choices in
the treatment of dyslipidemia, widely used and
commercially available on the global market (Merćep
et al., 2023). Statin derivatives work by inhibiting 3hydroxy-3-methyl-glutaryl-coenzyme reductase in a
competitive and very effective way to reduce the total
cholesterol levels, and achieve low density lipoprotein
levels, very low density lipoprotein levels, and
triglycerides levels, as well as being able to increase
high density lipoprotein levels in dyslipidemia patient
(Gesto et al., 2020).
Pharmaceutical
preparations
containing
simvastatin, atorvastatin, and rosuvastatin in tablet
form are produced by the pharmaceutical industry and
are found on the market. Tablets on the market must

meet the quality pharmacopoeia requirements to
ensure the quality of drugs is suitable for public
consumption (Al-Muhsin et al., 2022). Good quality of
drug preparations will support the achievement of
expected therapeutic effects. Active pharmaceutical
ingredient substances are a requirement that must be
met to ensure the quality of the marketed drugs
(Chaachouay & Zidane, 2024). One of the quality
requirements is that the content contained must meet
the content requirements as stated in the
pharmacopoeia or other standard literature. The
importance of quality control and quality assurance
has to be supervised and ensure that the drugs
produced are in accordance with the intended use
(Mashuri et al., 2025).
The World Health Organization pays great
attention to the quality, safety, and efficacy of
medicines, especially of active ingredients contained
in the finished goods (Dey & Nagababu, 2022). In
drug manufacture, examination of active substances is

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a requirement that must be met to ensure the quality
of drug preparations. Good quality drug preparations
will support the achievement of expected therapeutic
effects (Sampathkumar & Kerwin, 2024). United States
Pharmacopoeia includes monographs of simvastatin,
atorvastatin, and rosuvastatin tablets by analyzing
levels of active substances in tablet preparations using
high-performance liquid chromatography (Shulyak et
al., 2021).
The utilization of high-performance liquid
chromatography methods in analysis has the
disadvantage of the high cost of mobile phase,
longer preparation time and the longer sample
preparation time, uses more expensive solvents,
land has longer analysis time (Vempatapu et al.,
2024). Development
of the Fourier transform
infrared spectrophotometric method is an alternative
method in pharmaceutical analysis for qualitative and
quantitative analysis. The Fourier transform infrared
spectrophotometric method uses small amounts of
chemicals in sample preparation and could be a
low-cost (cheap), environmentally friendly (green),
simple (easy), and rapid (fast) analytical method
(Nerdy et al., 2021).
Although commercially available statin tablet
preparations typically contain only a single active
compound, the development of an analytical method
capable of simultaneously determining simvastatin,
atorvastatin, and rosuvastatin is essential. This
approach facilitates the detection of crosscontamination and enhances regulatory compliance.
The unified method can also streamline quality control
operations and reduce the validation workload.
Finally, this single method provides analytical flexibility
for both current and future pharmaceutical research
and product development. Based on this, this research
aims to develop a simple and fast analysis method that
can be used to determine levels of three different statin
derivatives (simvastatin, atorvastatin, and rosuvastatin)
in tablet preparations.
EXPERIMENTAL SECTIONS
Materials and Methods
This research is a descriptive study to determine the
levels of simvastatin, atorvastatin, and rosuvastatin in
tablet formulations containing a single active
pharmaceutical ingredient. The tablet preparations
used were marketed tablet preparations with a dose of
20 mg single active pharmaceutical ingredient per
tablet. The procedure used in this study is a
modification of the research of Nerdy et al, (2023), AlShami et al. (2024), and Shadrack et al. (2025). This
study consisted of several stages, namely:
determination of specific wave number of simvastatin,
atorvastatin, and rosuvastatin; determination of
linearity, limit of detection, and limit of quantitation of
simvastatin,
atorvastatin,
and
rosuvastatin;
determination of simvastatin, atorvastatin, and
rosuvastatin in tablet preparations; accuracy and

precision
of
rosuvastatin.

simvastatin,

atorvastatin,

and

Materials

The materials used in this study are Simvastatin
(Merck), Atorvastatin (Merck), Rosuvastatin (Merck),
Chloroform (Merck), Water (Merck), Norpid® 20 mg
(Graha Farma), Renapid® 20 mg (Bernofarm),
Rechol® 20 mg (Pharos Indonesia), Litorcom® 20 mg
(Combiphar), Atofar® 20 mg (Pratapa Nirmala), and
Giventor® (Nufarindo).

Tools

The tools used in this study are: Analytical Balance
- Cubis 120 (Sartorius), Fourier Transform Infrared
Spectrophotometer - Zn Se Cary 630 (Agilent);
Absorbance Transmission Module - Dialpath (Agilent),
Glassware (Iwaki); Sonicator (Krisbow).

Software

The software used in this study are Micro Lab PC
(Agilent), Micro Lab Quant (Agilent), and Statistical
Product and Service Solutions (International Business
Machines).
Determination of Specific Wave Number of
Simvastatin, Atorvastatin, and Rosuvastatin
An amount of 800 mg of simvastatin, 800 mg of
atorvastatin, and 800 mg of rosuvastatin was carefully
weighed, inserted separately into a 20 mL
volumetric flask, added 15 mL chloroform, sonicated
until dissolved, diluted with chloroform to the mark
line, and shaken until homogeneously mixed (a
standard stock solution of 40.0 mg/mL was
obtained with each concentration of simvastatin,
atorvastatin, and rosuvastatin). An amount of 2.5 mL
of each simvastatin, atorvastatin, and rosuvastatin
standard stock solution was pipetted, inserted in
three separate 20 mL volumetric flasks, diluted
with chloroform to the mark line, and shaken
until homogeneously mixed (a standard solution
containing of simvastatin, atorvastatin, and
rosuvastatin with a concentration 5.0 mg/mL in
three separate volumetric flasks). The absorption
spectrum measurements were carried out on
chloroform
and
separate standard solutions
(separate solutions of simvastatin, atorvastatin, and
rosuvastatin)
by
Fourier
transform
infrared
spectrophotometer with 0.1 mm cell in the wave
numbers range between 4000 cm–1 to 650 cm–1.
Measurements were made with six replications. The
overlap of chloroform absorption spectrum and
separate standard solution absorption spectrum
was carried out. The specific wavelengths of
simvastatin, atorvastatin, and rosuvastatin were
determined by Micro Lab PC (Nerdy et al., 2023).
Determination of Linearity, Limit of Detection, and
Limit of Quantitation of Simvastatin, Atorvastatin, and
Rosuvastatin
An amount of 1.0 mL, 1.5 mL, 2.0 mL, 2.5 mL, 3.0
mL, 3.5 mL, and 4.0 mL of each simvastatin,
atorvastatin, and rosuvastatin standard stock solution

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Development of Fourier Transform Infrared Spectrophotometric
Was pipetted, mixed same volume of each
simvastatin, atorvastatin, and rosuvastatin standard
stock solution in five separate 20 mL volumetric
flasks, diluted with chloroform to the mark line,
and shaken until homogeneously mixed (a standard
solution containing a mixture of simvastatin,
atorvastatin, and rosuvastatin with a concentration
of 2.0 mg/mL of each compound for the first
volumetric flask, 3.0 mg/mL of each compound for
the first volumetric flask; 4.0 mg/mL of each
compound for the first volumetric flask; 5.0 mg/mL of
each compound for the first volumetric flask; 6.0
mg/mL of each compound for the first volumetric
flask; 7.0 mg/mL of each compound for the first
volumetric flask; 8.0 mg/mL of each compound for
the first volumetric flask). The absorption spectrum
measurements were carried out on chloroform and
mixture
standard
solutions
(various
series
concentration of simvastatin, atorvastatin, and
rosuvastatin)
by
Fourier
transform
infrared
spectrophotometer with 0.1 mm cell in the wave
numbers range between 4000 cm–1 to 650 cm–1.
Measurements were made with six replications. The
overlap of chloroform absorption spectrum and
mixture standard solutions absorption spectrum was
carried out. The regression equation, determination
coefficient, and correlation coefficient of simvastatin,
atorvastatin, and rosuvastatin were determined by
Micro Lab Quant with the height calculation and
area calculation by the specific wavelengths of
simvastatin, atorvastatin, and rosuvastatin. The height
and the area obtained were further calculated to
obtain the limit of detection and the limit of
quantitation (Shadrack et al., 2025).
Determination of Simvastatin, Atorvastatin, and
Rosuvastatin in Tablet Preparations
A total of 20 tablets of simvastatin (3 samples),
atorvastatin (2 samples), and rosuvastatin (1
sample) were separately weighed and separately
powdered. An amount of powders were separately
weighed equivalent to 50 mg of simvastatin,
atorvastatin, or rosuvastatin, inserted into a 10 mL
volumetric flask, added
5 mL
of chloroform,
sonicated then diluted with chloroform until the
mark line, filtered, removed 3 mL of the first
filtrate, and collected the filtrate (concentration of
simvastatin, atorvastatin, or rosuvastatin 5.0 mg/mL).
The absorption spectrum measurements were carried
out on separate sample solutions (separate solutions
of simvastatin, atorvastatin, or rosuvastatin) by
Fourier transform infrared spectrophotometer with
0.1 mm cell in the wave numbers range between
4000 cm–1 to 650 cm–1. Measurements were made
with six replications. The samples' absorption spectra
were recorded. The height, area, concentration, and
percentage
of
simvastatin,
atorvastatin,
or
rosuvastatin were obtained and determined by Micro
Lab PC (Nerdy et al., 2023).

Nerdy Nerdy, et al.

Determination of Accuracy and Precision of
Simvastatin, Atorvastatin, and Rosuvastatin
Validation of analytical methods developed with
accuracy and precision test parameters is done by
measuring
recovery
percentage
and relative
standard
deviation in three specific ranges,
namely: 80%, 100%, and 120% (4.0 mg/mL, 5.0
mg/mL, and 6.0 mg/mL). Validation of accuracy
parameters and precision parameters in each
specific range was done with three replications
and used the standard addition method (70% of
the samples and 30% of the standard). In the
standard addition method, the number of samples
is added with the standard.
The accuracy
parameter with the recovery percentage criterion
is expressed by the difference between the two
results of the analysis (before and after the
addition of the standard) compared to the actual
content (the standard added). Precision parameters
with
relative
standard
deviation criteria are
calculated from a set of recovery percentage values
(Shadrack et al., 2025).
Determination of Range and Specificity of Simvastatin,
Atorvastatin, and Rosuvastatin
Validation of analytical methods developed with
range test parameter is done by analyzing the
concentration range of all parameters from the
analytical method validation obtained (linearity,
limit of detection, limi t of quantitation, accuracy,
and precision). The range of the concentration for
validation of analytical methods is the narrow
concentration range that met all parameters of the
analytical
method
validation.
Validation
of
analytical methods developed with specificity test
parameter is done by measuring placebo test. The
placebo was prepared by weight and blend tablet
placebo in proportions matching a typical tablet
formulation contained microcrystalline cellulose as
filler or diluent 35.0%, lactose monohydrate as
filler or diluent 35.0%, pregelatinized starch as
Binder and Disintegrant 12.5%, corn starch as binder
and disintegrant 12.5%, silicon dioxide as glidant
2.5%, magnesium stearate as lubricant 2.5%. An
amount equivalent to 200 mg as an average of 1
tablet mass, weight 500 mg of pacebo mixture was
inserted into a 10 mL volumetric flask, added 5 mL
of chloroform, sonicated then diluted with chloroform
until the mark line, filtered, removed 3 mL of the first
filtrate, and collected the filtrate. The absorption
spectrum measurements were carried out on placebo
solutions
by
Fourier
transform
infrared
spectrophotometer with 0.1 mm cell in the wave
numbers range between 4000 cm–1 to 650 cm–1.
Measurements were made with six replications.
The placebo absorption spectra were recorded. The
height, area, concentration, and percentage of
placebo were obtained and determined by Micro Lab
PC (Al-Shami et al., 2024).

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RESULTS AND DISCUSSIONS
The study began with the determination of the
spectrum of chloroform solvents in the wave numbers
range between 4000 cm–1 to 650 cm–1. This was
followed by the measurement of the spectrum of
simvastatin in chloroform solvent with a concentration
5.0 mg/mL in the wave numbers range between 4000
cm–1 to 650 cm–1; spectrum of atorvastatin in
chloroform solvent with a concentration 5.0 mg/mL in
the range of wave numbers 4000 cm–1 to 650 cm–1,
spectrum of rosuvastatin in chloroform solvent with a
concentration 5.0 mg/mL in the range of wave
numbers 4000 cm–1 to 650 cm–1; and spectrum of the
mixture of simvastatin, atorvastatin, and rosuvastatin
in a chloroform solvent with a concentration of each
compound 5.0 mg/mL in the range of wave numbers
4000 cm–1 to 650 cm–1, respectively. The spectra
resulted from the measurement were processed by
Micro Lab PC and can be seen in Figure 1 for
simvastatin, Figure 2 for atorvastatin, Figure 3 for
rosuvastatin, and Figure 4 for mixture of simvastatin,
atorvastatin, and rosuvastatin.
Measurement of the spectrum of simvastatin in
chloroform obtained simvastatin absorption peaks,
which showed different in overlay between the
spectrum of simvastatin in chloroform against the
chloroform absorption spectrum close to the wave
number results in the literature of 1712.7 cm–1
(Hakiem et al., 2021) with absorption of 0.816

absorbance unit. Measurement of the spectrum of
atorvastatin in chloroform obtained atorvastatin
absorption peaks, which showed differences in overlay
between the spectrum of atorvastatin in chloroform
against chloroform absorption spectrum close to the
wave number results in the literature, 1660.5 cm–1
(Salazar-Barrantes et al., 2025) with absorption of
0.326
absorbance
unit. Measurement of the
spectrum of rosuvastatin in chloroform obtained
rosuvastatin absorption peaks, which showed different
in overlay between the spectrum of rosuvastatin in
chloroform against chloroform absorption spectrum,
close to the wave number results in the literature,
965.4 cm–1 (Adel et al., 2023), with absorption of
0.803 absorbance unit.
The absorption spectrum of the mixture of
simvastatin, atorvastatin, and rosuvastatin in
chloroform shows the same wave number when
compared to the wave number and absorption of the
absorption spectrum of each absorption spectrum
from simvastatin, atorvastatin, and rosuvastatin in
chloroform. The results can be seen in Figure 5. This
proved that simvastatin, atorvastatin and rosuvastatin
did not influence each other in the quantitation
process. This method is often interfered by the solvent
spectrum, so a careful analysis is needed to determine
the wave number for the analysis of target substances
that are not interfered by the solvent used in the
analysis (Pasieczna-Patkowska et al., 2025).

Figure 1. Overlay between the spectrum of chloroform (– red line) against the spectrum of simvastatin
in chloroform (– blue line) in the wave numbers range between 4000 cm–1 to 650 cm–1

Figure 2. Overlay between the spectrum of chloroform (– red line) against the spectrum of atorvastatin
in chloroform (– blue line) in the wave numbers range between 4000 cm–1 to 650 cm–1
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Development of Fourier Transform Infrared Spectrophotometric

Nerdy Nerdy, et al.

Figure 4. Overlay between the spectrum of chloroform (– red line) against the spectrum of rosuvastatin
in chloroform (– blue line) in the wave numbers range between 4000 cm–1 to 650 cm–1

Figure 5. Overlay between the spectrum of chloroform (– red line), simvastatin (– purple line),
atorvastatin (– blue line), rosuvastatin (– green line), against mixture of simvastatin, atorvastatin, and
rosuvastatin (– orange line) in the wave numbers range between 2000 cm–1 to 650 cm–1
The study continued with the determination of
linearity of the mixture of simvastatin, atorvastatin,
and rosuvastatin in chloroform by measurements
of the mixture of simvastatin, atorvastatin, and
rosuvastatin
in
chloroform with concentrations
varying from 2.0 mg/mL to 8.0 mg/mL.
Measurements were made in the wave numbers
range between 4000 cm–1 to 650 cm–1. The
spectrum obtained from chloroform as a solvent
and standard solution containing a mixture of
simvastatin, atorvastatin, and rosuvastatin in
chloroform was overlaid and determination of the
linearity and regression line equations were applied
with Micro Lab Quant application using calculations
of peak height and peak area. Figure 6 shows the
overlaid spectrum of chloroform as a solvent and
standard
solution
containing
a mixture of
simvastatin, atorvastatin, and rosuvastatin in
chloroform at various concentration ranges (2.0
mg/mL to 8.0 mg/mL).
The results showed that the values of the
determination coefficient (R2) obtained ranged from
0.99826 to 0.99964 with the values of the correlation
coefficient (R) obtained ranged from 0.99913 to
0.99982. The determination coefficient values
obtained from both peak height and peak area show

good results because they were greater than 0.99 (Du
et al., 2022; Agberien & Örmeci, 2020), which shows
that both peak height and peak area can be used to
determine the concentration of simvastatin,
atorvastatin, and rosuvastatin. The correlation
coefficient values obtained from both peak height and
peak area show good results because they were
greater than 0.99 (Toropova et al., 2025), which
shows that both peak height and peak area have a
linear correlation with the concentration of
simvastatin, atorvastatin, and rosuvastatin. The results
can be seen in Table 1.
Determination of simvastatin, atorvastatin, and
rosuvastatin levels in tablet preparations was carried
out on tablets that are on the market with three
simvastatin trademark tablet preparations with a
strength of 20 mg per tablet; two atorvastatin
trademarks tablet preparations with a strength of 20
mg per tablet; and one rosuvastatin trademark tablet
preparation with a strength of 20 mg per tablet. The
results of the determination of levels were carried out
with six replications. Table 2 below shows the
tabulation results of determination of the levels of
active
pharmaceutical
ingredients
in
tablet
preparations with trademarks containing simvastatin,
atorvastatin, and rosuvastatin.

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Figure 6. Overlaid spectrum of chloroform as a solvent and standard solution containing a mixture of
simvastatin, atorvastatin, and rosuvastatin in chloroform at various concentration ranges (2.0 mg / mL to 8.0
mg / mL)
Table 1. The results of the calculation of slope, intercept, regression equation, determination coefficient, and
correlation coefficient from determination of the simvastatin, atorvastatin, and rosuvastatin linearity by peak
height and peak area
Number

Compound

Height or
Area

Slope

Intercept

Regression
Equation

Determination
Coefficient
(R2)

Correlation
Coefficient
(R)

Height

0.16912 0.01850

Y = 0.16912 ×
X + 0.01850

0.99851

0.99926

2

Area

7.19301 1.62359

Y = 7.19301 ×
X + 1.62359

0.99827

0.99914

3

Height

0.03997 0.03123

Y = 0.03997 ×
X + 0.03123

0.99861

0.99930

4

Area

0.95064 1.33623

Y = 0.95064 ×
X + 1.33623

0.99964

0.99982

5

Height

0.10823 0.04294

Y = 0.10823 ×
X + 0.04294

0.99840

0.99920

Area

1.33756 3.06409

Y = 1.33756 ×
X + 3.06409

0.99826

0.99913

1
Simvastatin

Atorvastatin

Rosuvastatin
6

Table 2. The results of determination of the levels of active pharmaceutical ingredients in tablet preparations
with trademarks containing simvastatin, atorvastatin, and rosuvastatin
Number
1
2
3
4
5
6

Compound
Simvastatin
Atorvastatin
Rosuvastatin

Sample
Norpid®
Renapid®
Rechol®
Litorcom®
Atofar®
Giventor®

Levels of Active Pharmaceutical Ingredients
Peak Height
Peak Area
102.83% ± 0.33%
102.75% ± 0.32%
101.92% ± 0.29%
101.11% ± 0.30%
99.56% ± 0.42%
99.39% ± 0.41%
101.81% ± 0.30%
101.99% ± 0.31%
100.38% ± 0.55%
100.56% ± 0.57%
102.65% ± 0.47%
102.51% ± 0.45%

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Nerdy Nerdy, et al.

Table 3. The results of validation with parameters of accuracy, precision, limit of detection, limit of quantitation,
and range.
Number

Parameter

1

Recovery
Relative Standard
Deviation
Limit of Detection
Limit of Quantitation
Range

2
3
4
5

Simvastatin
Height
Area
99.64%
99.47%

Results
Atorvastatin
Height
Area
99.55%
99.22%

Rosuvastatin
Height
Area
99.80%
99.99%

0.19%

0.28%

0.13%

0.21%

0.31%

0.14427
0.43717

0.15561 0.13969 0.07128 0.14972
0.47154 0.42332 0.21601 0.45370
4.0 mg/mL to 6.0 mg/mL

0.15621
0.47335

The determination of the levels of simvastatin,
atorvastatin, and rosuvastatin on the commercially
available trademark tablet preparations calculated by
using peak height and peak area. The determination
results was compared by t-test and obtained 0.42726
for the p-value. The p-value obtained is not less than
than 0.05 indicating that the determination results
calculated based on peak height and peak area are
not significantly different (Hawi et al., 2025). So that
the determination of the levels of simvastatin,
atorvastatin, and rosuvastatin can be done by
calculation of peak height or peak area calculation.
The results obtained in the content determination
based on the peak height analysis obtained the
results from 99.56% to 102.83%. The results obtained
in the content determination based on the peak area
analysis obtained the results from 99.39% to
102.75%. These results also indicate that the content
of simvastatin, atorvastatin, and rosuvastatin in
marketed tablet preparations fulfill the content
requirements for active pharmaceutical ingredient
assay, which ranges from 90.00% to 110.00% (PykaPająk, 2024).
The analytical method for determination the levels
of simvastatin, atorvastatin, and rosuvastatin that is
developed must be validated to prove that the
analysis method gives valid results. Table 3 shows
the tabulated results of validation with parameters
of accuracy, precision, limit of detection, limit of
quantitation, and range.
The validation of the analytical method for
determining the levels of simvastatin, atorvastatin,
and rosuvastatin in tablets with trademark
circulating on the market shows results that meet
the requirements. Validation of the analytical
method with the accuracy parameter by recovery
obtained results in the range of 99.22% to
99.99%; the results obtained meet the accuracy
test requirements, the recovery is in the range of
98.00% to 102.00% (Amin et al., 2024). Validation
of
the
analytical methods with the precision
parameter by relative standard deviation obtained
results in the range of 0.13% to 0.31%; the results
obtained meet the precision test requirements, the
relative standard deviation is not more than 2.00%
(Kotani et al., 2024).

0.25%

Validation of the analytical method with the limit of
detection parameter obtained results in the range of
0.07128 mg/mL to 0.15621 mg/mL; the results
obtained show good results with a small enough value
(32.01 fold to 70.14 fold lower than target
concentration 5.0 mg/mL) so that it can be detected
with low concentrations. Validation of the analytical
method with the limit of quantitation parameter
obtained results in the range of 0.21601 mg/mL to
0.47335 mg/mL; the results obtained show
satisfactory results with a small enough value so that it
can be quantified with low concentrations (10.56 fold
to 23.15 fold lower than target concentration 5.0
mg/mL). The value obtained for limit of detection and
limit of quantitation showed results that were much
lower than the target concentration. These results
showed quite satisfactory results (Gegenschatz et al.,
2022; Cravotto et al., 2022)
The results of the analytical method validation
include linearity at concentrations of 2.0 mg/mL to
8.0 mg/mL; accuracy at concentrations of 4.0
mg/mL to 6.0 mg/mL; precision at concentrations
of 4.0 mg/mL to 6.0 mg/mL; detection limit at
concentrations of 0.07128 mg/mL to 0.15621
mg/mL; quantitation limit at concentrations of
0.21601 mg/mL to 0.47335 mg/mL. The
combination of all parameters from the analytical
method validation obtained a range which the
concentration of 4.0 mg/mL to 6.0 mg/mL met all
parameters of the analytical method validation. In
analytical method validation, the range is the span
between the highest and lowest concentrations of the
analyte in a sample that the method has been shown
to measure with accuracy and precision (BlancoGarcía et al., 2025).
This research contributes to science by developing
an analytical method by fourier transform infrared
spectrophotometric which is very easy for sample
preparations and a very fast sample measurement
(Tkachenko & Niedzielski, 2022). However, the
spectrophotometric method has a weakness in terms
of separation and only background correction (Maia
et al., 2023). Simple liquid solvent extraction still
allows the matrix of pharmaceutical excipients
contained
in the tablet preparations extracted
together with the active pharmaceutical ingredient

209

Molekul, Vol. 20. No. 2, July 2025: 203 – 212
at the extraction stage during sample preparations
and also to be analyzed at the measurement stage in
infrared spectra. So it is necessary to select a very
selective s olvent in quantitative analysis (Kassouf et
al, 2024).
Non selective solvent will cause an overestimate to
the active pharmaceutical ingredients in the tablet
preparations
on
spectrophotometric
analysis
(Nikolaychuk, 2023). Placebo analysis is done to
ensure that the additional ingredients used in the
tablet formulation process do not interfere in the active
pharmaceutical ingredient analysis process (Gabel et
al., 2023, 2016). So it is very important for the
pharmaceutical industry to apply the simvastatin,
atorvastatin, and rosuvastatin analysis methods in
tablet preparations to conduct a placebo analysis at
the initial stage of the analysis. As an alternative to
increasing selectivity, solid phase extraction can also
be used, followed by validation of analytical methods
(Ashwini et al., 2025)
The developed method is able to analyze three
anti-dyslipidemia compounds derived from statins,
simvastatin, atorvastatin, and rosuvastatin. The
developed method has also been validated by
providing satisfaction using several analytical
method validation parameters, that is, accuracy,
precision, linearity, range, limit of detection, limit
of quantitation, and specificity. So that this method
can be an alternative valid method of reference
for the pharmaceutical industry and drug regulatory
authorities as a low cost (cheap), environmentally
friendly (green), simple (easy), and rapid (fast)
analysis method in determination of the levels of
simvastatin, atorvastatin, and rosuvastatin in tablet
preparations.
CONCLUSIONS
The developed Fourier transform infrared
spectrophotometric method has been successfully
applied to the analysis of simvastatin, atorvastatin,
and rosuvastatin in tablet formulations containing a
single active pharmaceutical ingredient. The
developed
Fourier
transform
infrared
spectrophotometric method was valid and met
validation parameters including accuracy, precision,
linearity, detection limit, quantitation limit, range, and
specificity.
CONFLICT OF INTEREST
The authors declare that there is no conflict of
interest in this research and this article.
ACKNOWLEDGMENTS
The researcher expresses gratitude to Universitas
Tjut Nyak Dhien for Research Fund, and also
expresses gratitude to Mutiara Mukti Farma Industri
Farmasi for the support of facilities for the tools and
materials used in this study

REFERENCES
Adel, I.M., ElMeligy, M.F., Amer, M.S., and Elkasabgy,
N.A.
(2023).
Polymeric
nanocomposite
hydrogel scaffold for jawbone regeneration the role of rosuvastatin calcium loaded silica
nanoparticles.
International
Journal
of
Pharmaceutics - X, 6, 100213. doi: 10.1016/
j.ijpx.2023.100213
Agberien, A. V., and Örmeci, B. (2020). Monitoring of
cyanobacteria
in
water
using
spectrophotometry and first derivative of
absorbance. Water, 12(1), 124. doi: 10.3390/
w12010124
Al-Muhsin, A., Ahad, A., Bin Jardan, Y.A., Raish, M.,
Ahmad, A., Alkharfy, K.M., and Al-Jenoobi, F.I.
(2022). Quality assessment of different brands
of atorvastatin tablets available in Riyadh, Saudi
Arabia. BMC Pharmacology and Toxicology,
23, 69. doi: 10.1186/s40360-022-00598-y
Al-Shami, N., Naseef, H., Moqadi, R., and Kanaze, F.
(2024). HPLC method development and
validation for the determination of apixaban
and
clopidogrel
in
novel
fixed-dose
combination tablets. Journal of Chemistry,
2024,
2675736.
doi:
10.1155/2024/
2675736
Amin, K.F.M., Obaydo, R.H., and Lotfy, H.M. (2024).
Chemometric
assisted spectrophotometric
methods for simultaneous drug determination
in new helicobacter pylori treatment regimens environmental
sustainability
assessment.
Sustainable Chemistry and Pharmacy, 42,
101849. doi: 10.1016/j.scp.2024.101849
Ashwini, T., Garg, S., Shenoy, P.A., Chandrashekhar,
R., Nayak, Y., and Nayak, U.Y. (2025) A
Quality
by
design
approach
with
comprehensive green analytical chemistry
assessment - development, validation, and
application of a high performance liquid
chromatographic method for quantifying
meropenem trihydrate in nanosponges and
marketed formulations. Journal of Applied
Pharmaceutical Science, 15(6), 70-95. doi:
10.7324/JAPS.2025.236517
Blanco-García, L., Cabarcos-Fernández, P., ÁlvarezFreire, I., Tabernero-Duque, M. J., MoredaPiñeiro, A., & Bermejo-Barrera, A. M. (2025).
Air-Assisted
Liquid–Liquid
Microextraction
(AALLME) as an alternative sample pretreatment for isolating tetrahydrocannabinol
(THC) from hair. Chemosensors, 13(6), 207.
doi: 10.3390/chemosensors13060207
Chaachouay, N., and Zidane, L. (2024). Plant-derived
natural products: a source for drug discovery
and development. Drugs and Drug Candidates,
3(1), 184-207. doi: 10.3390/ddc3010011
Cravotto, C., Fabiano-Tixier, A.-S., Claux, O., AbertVian, M., Tabasso, S., Cravotto, G., and
Chemat, F. (2022). Towards substitution of

210

Development of Fourier Transform Infrared Spectrophotometric
hexane as extraction solvent of food products
and ingredients with no regrets. Foods, 11(21),
3412. doi: 10.3390/foods11213412
Dey, S., and Nagababu, B.H. (2022). Applications of
food color and bio preservatives in the food and
its effect on the human health. Food Chemistry
Advances, 1, 100019. doi: 10.1016/j.focha.
2022.100019
Du, Y., Wang, Q., Yang, G., and Han, F. (2022).
Determination of 43 pesticide residues in intact
grape berries (Vitis vinifera L.) by using an
ultrasound assisted acetonitrile extraction
method followed by LC–MS/MS. Food Control,
140, 109123. doi: 10.1016/j.foodcont.2022.
109123
Gabel, J., Gnegel, G., Kessler, W., Sacré, P.Y., Heide,
L. (2023). Verification of the active
pharmaceutical ingredient in tablets using a low
cost near infrared spectrometer. Talanta Open,
8, 100270. doi: 10.1016/j.talo.2023.100270
Gegenschatz, S.A., Chiappini, F.A., Teglia, C.M., de
la Peña, A.M., and Goicoechea, H.C. (2022).
Binding the gap between experiments, statistics,
and method comparison - a tutorial for
computing limits of detection and quantification
in univariate calibration for complex samples.
Analytica Chimica Acta, 1209, 339342. doi:
10.1016/j.aca.2021.339342
Gesto, D. S., Pereira, C. M. S., Cerqueira, N. M. F. S.,
& Sousa, S. F. (2020). An atomic-level
perspective of HMG-CoA-reductase: The target
enzyme
to
treat
hypercholesterolemia.
Molecules, 25(17), 3891. doi: 10.3390/
molecules25173891
Hakiem, A.F.A., Mohamed, N.A., and Ali, H.R.H.
(2021). FTIR spectroscopic study of two
isostructural statins - simvastatin and lovastatin
as authentic and in pharmaceuticals.

Spectrochimica Acta Part A - Molecular and
Biomolecular Spectroscopy, 261, 120045. doi:

10.1016/j.saa.2021.120045
Hawi, S., Goel, S., Kumar, V., Giusca, C., Pearce, O.,
and Ayre, W.N. (2025). Precision laser
manufacturing and metrology of nature
inspired bioactive surfaces for antibacterial
medical implants. Surfaces and Interfaces, 62,
106267. doi: 10.1016/j.surfin.2025.106267
Kassouf, N., Zappi, A., Monticelli, M., & Melucci, D.
(2024). Analysis of solid formulates using UVvisible diffused reflectance spectroscopy with
multivariate data processing based on net
analyte signal and standard additions method.
Chemosensors, 12(11), 227. doi: 10.3390/
chemosensors12110227.
Kotani, A., Watanabe, R., Hayashi, Y., Machida, K.,
and Hakamata, H. (2024). Statistical reliability
of a relative standard deviation of
chromatographic peak area estimated by a
chemometric tool based on the FUMI theory.

Nerdy Nerdy, et al.

Journal of Pharmaceutical and Biomedical
Analysis, 237, 115777. doi: 10.1016/j.jpba.

2023.115777
Maia, R.N.A., Mitra, S., and Baiz, C.R. (2023).
Extracting accurate infrared lineshapes from
weak vibrational probes at low concentrations.
MethodsX, 11, 102309. doi: 10.1016/j.mex.
2023.102309
Mashuri, Y.A., Hasanah, M., Rahayu, I.D., Liverani,
M., Probandari, A., Batura, N., Schierhout, G.,
Ferdiana, A., Wulandari, L.P.L., Dewi, B.K., Jan,
S., Yeung, S., Wiseman, V., Day, R., and
Wibawa, T. (2025). Post market quality
assessment of antibiotics - findings from a
cross- sectional study using standardised
patients in Tabalong and Bekasi Districts,
Indonesia. BMJ Open, 15, e087801. doi:
10.1136/ bmjopen-2024-08780P
Merćep, I., Vujević, A., Strikić, D., Radman, I., Pećin,
I., & Reiner, Ž. (2023). Present and future of
dyslipidaemia treatment - A review. Journal of
Clinical Medicine, 12(18), 5839. doi:
10.3390/jcm12185839
Nerdy, N., Margata, L., Sembiring, B. M., Ginting, S.,
Putra, E. D. L., and Bakri, T. K. (2021).
Validation of the developed zero order infrared
spectrophotometry method for qualitative and
quantitative analyses of tranexamic acid in
marketed tablets. Molecules, 26(22), 6985. doi:
10.3390/molecules26226985
Nerdy, N., Margata, L., Zebua, N.F., Lestari, P., Bakri,
T.K., Yusuf, F., and Aulianshah, V. (2023).
Development and validation of fast and simple
fourier transform infrared spectrophotometric
method for analysis of thiamphenicol in capsule
dosage form. Science and Technology
Indonesia, 8(3), 344-352. doi: 10.26554/sti.
2023.8.3.344-352
Nikolaychuk, P. A. (2023). UV-Spectrophotometric
Determination of the active pharmaceutical
ingredients meloxicam and nimesulide in
cleaning validation samples with sodium
carbonate. Multidisciplinary Scientific Journal,
6(2),
248-266.
https://doi.org/10.3390/
j6020019
Pasieczna-Patkowska, S., Cichy, M., and Flieger, J.
(2025). Application of Fourier transform
infrared (FTIR) spectroscopy in characterization
of green synthesized nanoparticles. Molecules,
30(3), 684. doi: 10.3390/molecules30030684
Pyka-Pająk, A. (2024). Determination of the active
pharmaceutical ingredients in Saridon tablets
using an economical and sensitive thin layer
chromatography method combined with
densitometry. Analytica, 5(1), 1-16. doi:
10.3390/analytica5010001
Sadowska, A., Osiński, P., Roztocka, A., KaczmarzChojnacka, K., Zapora, E., Sawicka, D., and
Car, H. (2024). Statins - from fungi to

211

Molekul, Vol. 20. No. 2, July 2025: 203 – 212
pharmacy. International Journal of Molecular
Sciences, 25(1), 466. doi: 10.3390/ijms250
10466
Salazar-Barrantes, K.A., Abdala-Saiz, A., VegaBaudrit, J.R., Navarro-Hoyos, M., and ArayaSibaja, A.M. (2025). Formulation and
evaluation of atorvastatin calcium trihydrate
form I tablets. PLoS ONE, 20(2), e0317407.
doi: 10.1371/journal.pone.0317407
Sampathkumar, K., and Kerwin, B.A. (2024).
Roadmap for drug product development and
manufacturing of biologics. Journal of
Pharmaceutical Sciences, 113(2), 314-331.
doi: 10.1016/j.xphs.2023.11.004
Shadrack, R.S., Kotra, K.K., Tari, D., Tabi, H., Botleng,
J., Kelep, R., and Regenvanu, L. (2025). A Rapid
Method for Methanol Quantification in Spirits
Using UV Visible Spectroscopy and FTIR. PeerJ
Analytical Chemistry, 7, e35. doi: 10.7717/
peerj-achem.35
Shulyak, N., Piponski, M., Kovalenko, S., Bakovska
Stoimenova, T., Balkanov, T., El-Subbagh, H. I.,

Drapak, I., Omotosho, J. O., and Logoyda, L.
(2021). Development of a novel, fast, simple
HPLC method for determination of atorvastatin
and its impurities in tablets. Scientia
Pharmaceutica, 89(2), 16. doi: 10.3390/
scipharm89020016
Tkachenko, Y., and Niedzielski, P. (2022). FTIR as a
method for qualitative assessment of solid
samples in geochemical research: A review.
Molecules, 27(24), 8846. doi: 10.3390/
molecules27248846
Toropova, A. P., Toropov, A. A., Roncaglioni, A., &
Benfenati, E. (2025). Using the coefficient of
conformism of a correlative prediction in
simulation of cardiotoxicity. Toxics, 13(4), 309.
doi: 10.3390/toxics13040309
Vempatapu, B.P., Kumar, J., Upreti, B., and Kanaujia,
P.K. (2024). Application of high performance
liquid chromatography in petroleum analysis challenges and opportunities. TrAC Trends in
Analytical Chemistry, 177, 117810. doi:
10.1016/j.trac.2024.117810

212