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UV-VIS Determination of Acetylsalicylic Acid in Aspirin Tablets Using

Different Solvents and Conditions

Chapter · January 2020

DOI: 10.1007/978-3-030-17971-7_85

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 UV-VIS Determination of Acetylsalicylic Acid
in Aspirin Tablets Using Different Solvents
and Conditions
M. Dacić, A. Uzunović, A. Kunić, S. Pilipović, and A. Šapčanin
Abstract
Acetylsalicylic acid is the salicylic drug which is
commonly used like analgesic, antipyretic and
anti-inflammatory drug. There are a lot of methods for
analysis, but we decided for UV-VIS spectroscopy. In this
paper are correlation results from UV-VIS determination
of acetylsalicylic acid in six aspirin tablets from different
manufacturers in Bosnia and Herzegovina using two
different methods and changing parameters in first one.
Standards and samples are prepared at the same way. First
method was done using 1 M NaOH for degradation of
acetylsalicylic acid and development of colors with
0.02 M FeCl3 in 0.03 HCl (optimal pH = 1.6). For this
analysis we made (multiple times) five standards with
different concentrations (50–250%), changing pH of
FeCl3/HCl solution (0.60, 1.60, 1.95). Then, we changed
the concentration range of standards: 50–250, 80–120,
80–160%. The best absorbance results were at 530 nm
using FeCl3/HCl solution at pH 1.60, so we analyze
samples using that solutions. The second method was
dissolving standards and samples in concentrated ethanol
and measuring absorbance at 227 nm. Both method gave
good results and can be used for pharmaceutical research.



Keywords
Acetylsalicylic acid UV-VIS spectroscopy
Absorbance
M. Dacić
A. Kunić (&)
Faculty of Pharm and Health, Slavka Gavrančića 17c, Travnik,
Bosnia and Herzegovina
e-mail: mineladaca@gmail.com
A. Uzunović
S. Pilipović
Agency for Medicinal Products and Medical Devices, Titova 9,
Sarajevo, Bosnia and Herzegovina
A. Šapčanin
Faculty of Pharmacy, University of Sarajevo, Zmaja od Bosne 8,
Sarajevo, Bosnia and Herzegovina
© Springer Nature Switzerland AG 2020
A. Badnjevic et al. (eds.), CMBEBIH 2019, IFMBE Proceedings 73,
https://doi.org/10.1007/978-3-030-17971-7_85
1 Introduction
Acetylsalicylic acid is the prototypical analgesic used in the
treatment of mild to moderate pain. It has anti-inflammatory
and antipyretic properties and acts as an inhibitor of
cyclooxygenase which results in the inhibition of the
biosynthesis of prostaglandins. Aspirin also inhibits platelet
aggregation and is used in the prevention of arterial and
venous thrombosis [1].
Acetylsalicylic acid appears to produce analgesia by
virtue of both a peripheral and CNS effect. Peripherally,
acetylsalicylic acid acts by inhibiting the synthesis and
release of prostaglandins. Acting centrally, it would appear
to produce analgesia at a hypothalamic site in the brain,
although the mode of action is not known. Acetylsalicylic
acid also acts on the hypothalamus to produce antipyresis;
heat dissipation is increased as a result of vasodilatation and
increased peripheral blood flow. Acetylsalicylic acid’s anti-
pyretic activity may also be related to inhibition of synthesis
and release of prostaglandins [2].
There are a lot of methods which are used for determi-
nation of acetylsalicylic acid, like HPLC, UV-VIS determi-
nation, volumetric etc. For this paper, we decided to use
UV-VIS spectroscopy because of simplicity and low costing.
UV-visible spectroscopy is a technique that readily
allows one to determine the concentrations of substances and
therefore enables scientists to study the rates of reactions,
and determine rate equations for reactions, from which a
mechanism can be proposed [3].
For the analysis, we took six acetylsalicylic acid tablets
from local market. In this paper this tablet will be designated
as Sample 1–6.
563
564
2 Materials and Methods
2.1 Standard Preparation
2.1.1 Method with FeCl3/HCl Solution
The method is based at the procedure described at Maksi-
mović [4]. For standards preparation, we use USP Acetyl-
salicylic acid Reference Standard (500 mg) CAT
No. 1044006. At the analytical balance (Mettler Toledo,
±0.01 mg) was measured 10.31 mg of standard in the vol-
umetric flask of 10 ml and added 1 ml 1.0 M NaOH, then
heated to boiling and boiled five minutes. The flask was
cooled and added distillated water to the mark. That was the
stock solution which we used for making diluted standards
in volumetric flask of 10 ml. We made three different series
of five standards with different concentrations and different
pH. There was pipetted appropriate amount of stock stan-
dards in the volumetric flask of 10 ml depend of concen-
tration and diluted with 0.02 M FeCl3 in 0.03 M HCl (for
three series of standards used FeCl3/HCl solution had three
different pH). pH of FeCl3/HCl solution was set using NaOH
and HCl, depend of required values of pH, which was: 0.60,
1.60 and 1.95.
2.1.2 Method with Ethanol
For standards preparation, we used the same Acetylsalicylic
acid Reference Standard. At the analytical balance (Mettler
Toledo, ±0.01 mg) was measured 5.00 mg of standard in
volumetric flask of 10 ml and added ethanol to the mark.
After dissolving, we pipetted 0.4, 0.5, 0.6, 0.7 and 0.8 ml of
stock solution in volumetric flask of 10 ml and added dis-
tillated water to the mark, so we had five diluted standards
with concentrations: 20, 25, 30, 35 and 40 µg/ml (80, 100,
120, 140, 160% respectively).
2.2 Sample Preparation
2.2.1 Method with FeCl3/HCl Solution
From each batch, we took 20 tablets using random selection
and measure the tablets mass for calculating average mass of
the tablet. Then, the tablets were crumbled and weight the
mass of tablet powder that suits the weight of 50.00 mg
acetylsalicylic acid in volumetric flask of 100 ml. In flask
was added 5 ml 1 M NaOH and heated to the boiling and
boiled five minutes. When the flask was cooled, we added
distillated water to the mark and mixing. Solution was fil-
tered and 0.25 ml of filtrate was pipetted in volumetric flask
of 10 ml and added FeCl3/HCl solution (pH = 1.60) to the
mark. That was the solution for measuring the absorption at
530 nm with FeCl3/HCl as reference solution.
M. Dacić et al.
2.2.2 Method with Ethanol
There was measured the mass of the tablet powder that suits
the weight of 5.00 mg acetylsalicylic acid in volumetric
flask of 10 ml and added ethanol to the mark. After dis-
solving, we filtrated the solution and pipette 0.50 ml of fil-
trate in volumetric flask of 10 ml and dilute with distillated
water to the mark. That was the solution for measuring the
absorption at 227 nm with distillated water as reference
solution.
UV-VIS spectroscopy. Analysis was carried out using a
double beam UV-VIS spectrophotometer Shimatzu,
UV-1800, using quartz cells and Shimatzu software at 227
and 530 nm.
Quantitation. The calibration curve was used with different
concentrations. The concentration ranges of the standard
curves were 10–50, 40–60 and 20–40 µg/ml. For quantita-
tion acetylsalicylic acid in tablets was used curve with
concentration range 20–40 µg/ml.
Recovery study. In order to verify the accuracy and preci-
sion of the analytical procedure, recovery studies were car-
ried out by calculating the amount of the measured mass
(mm) through the determined mass (md) of acetylsalicylic
acid (Formula 1).
R% ¼ mm =md  100 ð1Þ
There was calculated standard deviation and relative
standard deviation which was under 2.0 in some of analyses,
so accuracy and precision was satisfactory recording USP
parameters [5]. The problem was in absorbance response at
the lower pH, which is the reason why we used the FeCl3/
HCl solution with the pH = 1.60 (better absorbance
response).
3 Results and Discussion
3.1 Method with FeCl3/HCl Solution
Calibration Curves
At the beginning we used FeCl3/HCl solution with pH 0.60
to development the colors of standards with concentration
range 50–250%. Table 1 are results of determined amount of
acetylsalicylic acid in standards.
From the recovery we can realize that the method isn’t
good, because we didn’t find the amount which we added.
At Fig. 1 is showed calibration curve of correlation of
concentration-absorbance.
V-VIS Determination of Acetylsalicylic Acid … 565

Table 1 Correlation of added and measured mass of acetylsalicylic acid standards at concentration range 50–250%, pH = 0.60
% µg/mL Added (mg) Measured (mg) R (%)
50 9.994 24.985 41.535 166.240
100 19.988 49.970 49.970 100.000
150 29.982 74.955 57.862 77.196
200 39.976 99.940 65.588 65.628
250 49.970 124.925 74.716 59.809

y = 0,00026x + 0,05312 y = 0,00070x + 0,04324

Calibration curve R² = 0,99925
Calibration curve R² = 0,99774
0,13 0,13

0,11 0,12

Abs
Abs

0,09 0,11
0,07 0,1
0,05 0,09
0 50 100 150 200 250 300 70 80 90 100 110 120 130
% %

Fig. 1 Calibration curve %-absorbance at the concentration range

50–250%, pH = 0.60 Fig. 2 Calibration curve %-absorbance at the concentration range
80–120%, pH = 0.60
Correlation coefficient (R2) is 0.99925 so the linearity is
good, but we can see that absorbance is weak and all stan- Table 3 Correlation of added and measured mass of acetylsalicylic
acid standards at concentration range 50–250%, pH = 1.60
dards had absorbance bellow 0.11 so at the pH 0.60 there
doesn’t arise the tetraaquasalicylic iron (III) complex which % µg/mL Added (mg) Measured (mg) R (%)
is responsible for color development. 50 10.31 5.155 6.066 117.663
In the following text are results of measured standards at 100 20.62 10.310 10.310 100.000
different concentrations and with different pH values. 150 30.93 15.465 14.788 95.624
All results of standards which have pH 0.60 had lower 200 41.24 20.620 19.224 93.227
absorbance and recovery wasn’t satisfying (see Table 2 and
250 51.55 25.775 23.501 91.177
Fig. 2). Standards with pH 1.60 had the best absorbance
values, so we decided to use FeCl3/HCl with pH 1.60 (see
Table 3 and Fig. 3) which is recommended by Maksimović
[4]. Standards with pH 1.95 had good measured absorbance R² = 0,99992
(see Table 4 and Fig. 4), but the correlation coefficient was 0,6
0.98583 so the curve linearity isn’t satisfactory for phar- 0,4
Abs

maceutical researching where has to be >0.995 [5].

For the final concentrations, we made standards with 0,2
concentration range 80–160%, pH 1.60 (see Table 5 and 0
Fig. 5). 0 50 100 150 200 250 300
%

Table 2 Correlation of added and measured mass of acetylsalicylic

acid standards at concentration range 80–120%, pH = 0.60
% µg/mL Added (mg) Measured (mg) R (%) Fig. 3 Calibration curve %-absorbance at the concentration range
50–250, pH = 1.60
80 39.976 39.976 44.065 110.230
90 44.973 44.973 47.424 105.449
100 49.970 49.970 49.970 100.000
110 54.967 54.967 53.643 97.592 Recovery’s for this standards (Table 5) has values 95.53–
120 59.964 59.964 56.476 94.182 100% and SD and RSD is lower than 2.00 so we can use this
curve for pharmaceutical researching.
566 M. Dacić et al.

Table 4 Correlation of added and measured mass of acetylsalicylic UV-VIS determination of aspirin
acid standards at concentration range 50–250%, pH = 1.95 102
99,2058 99,4264

% aspirin amount
% µg/mL Added (mg) Measured (mg) R (%) 100 98,3857 98,2841
98 97,3378
50 10.31 5.155 6.397 124.097
96 94,653
100 20.62 10.310 10.310 100.000 94
150 30.93 15.465 15.314 99.023 92
90
200 41.24 20.620 21.004 101.865 1 2 3 4 5 6
250 51.55 25.775 23.213 90.059 Samples

Fig. 6 Results of measured % of acetylsalicylic acid in aspirin tablets

Calibration curve y = 0,00220x + 0,04837

R² = 0,98583 Sample Results
0,7
For this paper, we analyzed six tablet samples from six
0,6
Abs (535 nm)

0,5 different manufacturers. Results are in Fig. 6.

0,4 For pharmaceutical determination, the amount of acetyl-
0,3 salicylic acid in aspirin tablets have to be 90–110% (EU
0,2 Pharmacopoeia, Monograph of aspirin tablets and USP,
0,1
Aspirin tablets monograph) [6, 7]. The measured amounts
0
0 50 100 150 200 250 300 were 94.65–99.43% so all analyzed tablets has satisfying
% amount of acetylsalicylic acid.

Fig. 4 Calibration curve %-absorbance at the concentration range

50–250, pH = 1.95
3.2 Method with Ethanol
Table 5 Correlation of added and measured mass of acetylsalicylic
acid standards at concentration range 80–160%, pH = 1.60 Calibration Curves
% µg/mL Added (mg) Measured (mg) R (%) We made one calibration curve following the method which
80 20.620 8.248 8.130 98.570 is described earlier. Results are in Table 6.
100 25.775 10.310 10.310 100.000 In Table 6 are results which shows that recovery, SD and
120 30.930 12.372 12.076 97.607
RSD for acetylsalicylic acid standards, which are prepared in
ethanol, has excellent values, so we used calibration curve
140 36.085 14.434 13.874 96.117
from this results for determination acetylsalicylic acid in
160 41.240 16.496 15.759 95.531
tablets (see Fig. 7).
AVG 97.565 From the R2, which is 0.99989, we can realize that the
SD 1.8148 linearity is good for pharmaceutical researching, so we use
RSD 1.860 this curve for analyzing samples.

Table 6 Correlation of added and measured mass of acetylsalicylic

acid standards
y = 0,00254x + 0,01986
Calibration curve R² = 0,99860 % µg/mL Added (mg) Measured (mg) R (%)
0,45
0,4
80 20 4 3.981 99.517
0,35 100 25 5 5.000 100.000
Abs

0,3 120 30 6 5.942 99.041

0,25 140 35 7 6.951 99.297
0,2 160 40 8 7.939 99.232
50 70 90 110 130 150 170
% AVG 99.417
SD 0.367
Fig. 5 Calibration curve %-absorbance at the concentration range
80–160%, pH = 1.60 RSD 0.370
V-VIS Determination of Acetylsalicylic Acid … 567

Calibration curve y = 0,00963x + 0,00831 R² = 0,99989 minutes in ultrasonic bath etc. are crucial factor for solving
1,6
samples, so have to be same to get good results.
1,4
1,2
Abs

1
4 Conclusion
0,8
0,6 There was analyzed acetylsalicylic acid at six tablet samples
50 70 90 110 130 150 170
%
with two different methods. The conditions and preparing
was different, but both methods gave good results. Method
Fig. 7 Calibration curve %-absorbance at the concentration range with FeCl3/HCl solution are sensitivier so all parameters has
80–160% to be right to get good results, so it is a little beat difficultier
than a method with ethanol. With second method is easier to
Table 7 Sample results analyze samples but at first step, the most important thing is
to make UV-VIS spectar of standard and make conclusion at
Sample Measured Manufacturer declaration %
(mg/tbl) (mg)
which wavelength is the best absorbanse response for stan-
dards, so you can use that wavelength for samples. Results
1 104.407 100 104.407
are good in both methods, so they can be used for phar-
2 97.299 100 97.299
maceutical researching.
3 99.004 100 99.004
4 97.367 100 97.367
5 98.553 100 98.553 References
6 97.425 100 97.425
1. PubChem, Open Chemistry Database, https://pubchem.ncbi.nlm.
nih.gov/compound/aspirin#section=Top. Last accessed 12.11.2018
2. DrugBank, https://www.drugbank.ca/drugs/DB00945. Last acce
Sample Results ssed 12.11.2018
3. RSC, Advanced the Chemical Sciences. http://www.rsc.org/
The same samples, which were analyzed with first method,
learnchemistry/content/filerepository/CMP/00/001/304/UV-Vis_Stu
are analyzed with method with ethanol too. Results are in dent%20resource%20pack_ENGLISH.pdf. Last accessed 12.11.
Table 7. 2018
Measured amount range for this tablets was 98.30– 4. Maksimović, M., Ćavar, S., Vidić, D.: Praktikum iz osnova
organske hemije, 73–83. Faculty of Sciences, Sarajevo (2009)
104.41%, which is within the EU and USP pharmacopoeias
5. UV-VIS spectroscopy, USP—Physical tests (857) UV-VIS spec-
range of 90–110%, so this method can be use for determi- troscopy, page: 40. https://hmc.usp.org/sites/default/files/docum
nation of acetylsalicylic acid too. The problems which we ents/HMC/GCs-Pdfs/c857.pdf. Last accessed 14.11.2018
had doing this method was related with quantitative solving 6. EU Pharmacopoeia, Monograph of aspirin tablets
7. USP monograph of aspirin tablets, https://www.uspnf.com/sites/
tablets. There is very important to have the same all condi-
default/files/usp_pdf/EN/USPNF/iras/aspirin_tablets.pdf. Last acces
tions in preparing standards and samples. Temperature, sed 15.11.2018

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