FT-IR Spectrophotometric analysis of acetylsalicylic acid and its

pharmaceutical formulations

Andrei A. Bunaciu1, Hassan Y. Aboul-Enein2* and Şerban Fleschin3

1
CROMATEC_PLUS SRL, Analytical Research Department, 18 Sos. Cotroceni, Bucharest - 6, 060114, Romania.
2
College of Pharmacy, University of Sharjah, P.O.Box 27272 , Sharjah, United Arab Emirates.
3
Department of Analytical Chemistry, Faculty of Chemistry, University of Bucharest, 92-96, Sos. Panduri, Bucharest-
5, 050663, Romania.

Received: April 28, 2006 Accepted (in revised form): July 5, 2006

Abstract Introduction

A Fourier transform infrared (FT-IR) spectrometric
method was developed for the rapid, direct measurement
of acetylsalicylic acid (ASA) in different pharmaceutical
products. Conventional KBr-spectra spectra were
compared for the best determination of active substance
in drug preparations. The Beer-Lambert law and two
chemometric approaches, partial least squares (PLS)
and principal component regression (PCR+) methods,
were used in data processing.
Keywords: FT-IR analysis, acetylsalicylic acid, drug
analysis, calibration curve, chemometric approaches
Résumé
Nous avons développé une méthode de spectrométrie
infrarouge transformée de Fourier (FT-IR) pour la
mesure rapide et directe de l’acide acétylsalicylique
(ASA) dans différents produits pharmaceutiques. Les
spectres KBr conventionnels ont été comparés pour une
meilleure détermination de la substance active dans la
préparation des médicaments. La loi de Lambert-Beer
et deux approches chémométriques, la méthode des
moindres carrés partiels (PLS) et celle de la composante
de régression principale (PCR+) ont servi au traitement
des données.
Infrared spectrometry (IR) provides a useful way for
the identification of drugs (1-4). However, the traditional
techniques employed to obtain the IR spectra, such as
alkali halides disks, mulls and thin films, are sometimes
not adequate for quantitative analysis. Fourier Transform
(FT-IR) permits continuous monitoring of the spectral
baseline and simultaneous analysis of different compo-
nents of the same sample (5,6).
Acetylsalicylic acid (ASA, o-acetoxybenzoic acid,
aspirin ) (see Figure 1) is one of the oldest analgetics
substance. Already Hippocrates (460-370 B.C.) used an
extract of bark of willow trees – containing a high quan-
tity of salicylic acid to relieve pain (7). Aspirin is used
to relieve mild to moderate pain, reduce fever, redness,
and swelling, and to help prevent blood from clotting.
It is used to relieve discomfort caused by numerous
medical problems, including headaches, infections, and
arthritis. It is also used to reduce the risk of a second
heart attack or stroke. Larger doses of aspirin are used
to treat gout. Acetylsalicylic acid inhibits the body’s pro-

*
Author to whom correspondence should be addressed:
e-mail: Professor H.Y. Aboul-Enein (hyaboulenein@yahoo.com) Figure 1. Structure of acetylsalicylic acid.

Canadian Journal of Analytical Sciences and Spectroscopy

254 Andrei A. Bunaciu, Hassan Y. Aboul-Enein, and Şerban Fleschin
duction of prostaglandins. Prostaglandins are hormone-
like substances which are involved in the regulations of
varied processes such as pain, fever, inflammations and
thromboses. Prostaglandins elicit signals of pain and so
by obstructing their synthesis, no pain can be felt.
The techniques described for the assay of ASA are
high performance liquid chromatography (8), and other
techniques relying spectrophotometry (9-12) or capillary
electrophoresis techniques (13).
The determination of the major components in drugs
with FT-IR spectrometry provides an enormous amount
of spectroscopic information of a sample. Chemomet-
ric methods, such as principal component regression
(PCR+), Improved Principal Component Regression,
and partial least squares (PLS2, Multicomponent Partial
Least Squares) analysis are commonly used to extract
the specific information relevant to the analyte of inter-
est from the full spectrum (1, 14). These two techniques
yields more accurate calibration models compared with
multiple linear regression (MLR) where a restricted set
of absorption bands is used in the calibration. A good
introduction to the partial least squares (Projection to
Latent Structures, PLS) method is given by Geladi and
Kowalski (15).
The purpose of the present study is to investigate
the potential of FT-IR spectrometry to quantify ASA in
pharmaceutical preparations. The proposed analytical
method in this study was performed on standard ASA
and three different aspirin containing pharmaceutical
formulations namely BUFFERIN - sample A containing
about 325 mg of ASA per tablet, AGGREX - sample B
containing about 75 mg of ASA per tablet and EUROPI-
RIN T - sample C containing about 500 mg of ASA per
tablet, respectively.
The main objective of this work was to develop a
chemometric procedure for the fast and accurate deter-
mination of ASA in commercial pharmaceutical formula-
tions by using the Beer-Lambert law and/or PCR+ and
PLS approaches, reducing the sample pre-treatment and
providing direct FTIR measurement.
Experimental
Apparatus
Data acquisition was performed using a Spectrum100
Systems FT-IR spectrometer equipped with Spectrum
for Windows v.5.01 (Perkin Elmer Co., Beaconsfield,
Bucks, UK). The commercial softwares used to gener-
ate analysis for the principal component analysis were
QUANT+ expert v.4.51 and Spectrum Beer’s law v.2.01
Volume 51, No. 5, 2006
(Perkin Elmer Co. UK).
Reagents and materials
Acetylsalicylic acid was provided by Leiner Healt
Prod. Inc. Carson, CA, USA. The pharmaceuticals
were manufactured by Bristol – Myers Squibb Egypt
a Bristol-Myers Squibb Company (New York- Cairo),
(sample A), from RAMEDA (The Tenth of Ramadan)
Co., Egypt (sample B), and from Europharm Braşov
Romania (sample C), respectively.
Recommended procedures
Conventional fused KBr disk spectra were recorded
between 4000 and 350 cm-1, by averaging 64 scans for
each spectrum with a resolution of 4 cm-1 (data point reso-
lution/interval cm-1) with a DTGS detector from samples
prepared by compressing 2.0 mg of drug sample with
spectral grade KBr, while the background was spectral
grade KBr (ASA_KBr). For calibration, the spectra were
recorded with the same detector from samples prepared
by compressing the standard substance, ASA, (0.5 mg,
1.0 mg, 1.5 mg and 2.0 mg, respectively) in spectral
grade KBr.
The universal ATR spectra were recorded between
4000 and 650 cm-1, by averaging 32 scans for each spec-
trum with a Spectrum 100 Universal Diamond/ZnSe ATR
with 3 Reflection Top-Plate (ASA_UATR). In order to
compress the sample against the crystal, a pressure plate
and clamp are provided.
Experimental parameters and calibration methods
(PCR+, PLS1, PLS2 or the Beer-Lambert law, respec-
tively) were compared and recommendations for the best
options in ASA analysis were made.
Results and Discussion
Figures 2-5 present spectra of standard acetylsalicylic
acid as well as for the pharmaceutical preparations using
the KBr disk method and UATR, respectively. It is of
interest to note that there are no significant differences
in the fingerprinting region between the spectra obtained
from the KBr disk and UATR methods.
Chemometric techniques which are known as nu-
merical techniques are useful for the spectrophotometric
resolution of complex mixtures of analytes without the
need of prior separation or extraction. Although both
PCR and PLS give succesuful results, they have several
disadvantages such as using abstract mathematical theory
and various softwares.
In PCR and PLS2, the spectra are modeled by one
FT-IR Spectrophotometric analysis of acetylsalicylic acid and its pharmaceutical formulations 255

Fig.2. FT-IR spectra of ASA standard using the KBr-disk method.

Figure 3. FT-IR spectra of ASA standard using the UATR method.

Canadian Journal of Analytical Sciences and Spectroscopy

256 Andrei A. Bunaciu, Hassan Y. Aboul-Enein, and Şerban Fleschin

Figure 4. FT-IR spectra of pharmaceutical preparations containing ASA using the KBr-disk. a. Bufferin 325mg ASA per tablet; b. Aggrex
75 mg ASA per tablet; c. Europirin 500mg ASA per tablet.

Figure 5. FT-IR spectra of pharmaceutical preparations containing ASA using the UATR method. a. Bufferin 325mg ASA per tablet; b.
Aggrex 75 mg ASA per tablet; c. Europirin 500mg ASA per tablet.

Volume 51, No. 5, 2006

FT-IR Spectrophotometric analysis of acetylsalicylic acid and its pharmaceutical formulations 257

Figure 6. Calibration curve according to the Beer-Lambert law for the quantitative determination of ASA in pharmaceutical preparations.

Canadian Journal of Analytical Sciences and Spectroscopy

258 Andrei A. Bunaciu, Hassan Y. Aboul-Enein, and Şerban Fleschin
Table 1. Comparison of the ASA determination in tablets obtained by FT-IR and the Beer-Lambert law methods.
Sample A
ASA_KBr ASA_UATR
Content (mg/tablet) 318.88 312.55
RSD (%) (n = 5) 3.25 3.99
Sample B
ASA_KBr ASA_UATR
Content (mg/tablet) 72.26 70.58
RSD (%) (n = 5) 3.35 4.05
Sample C
ASA_KBr ASA_UATR
Content (mg/tablet) 487.34 477.61
RSD (%) (n = 5) 3.14 4.11
Calibration parameters
Slope 15.5100
Intercept -1.2123
Correlation coefficient 0.9886

Table 2. Comparision of the ASA determination in tablets using the FT-IR chemometric approaches.
Sample A
ASA_KBr ASA_UATR
PCR+ PLS PCR+ PLS
RMS Error 0.2189A 0.2023A 0.1949A 0.2019A
Peak-to-Peak Error 0.9322A 0.9022A 0.9322A 0.8729A
Total M-Distance 0.927 0.953 0.967 0.892
Content (mg/tablet) 312.25 315.87 316.34 310.68
RSD (%) (n = 5) 2.15 2.07 2.85 2.93
Sample B
ASA_KBr ASA_UATR
PCR+ PLS PCR+ PLS
RMS Error 0.09019A 0.07863A 0.06929A 0.07129A
Peak-to-Peak Error 0.4232A 0.3572A 0.3312A 0.3359A
Total M-Distance 0.952 0.848 0.929 0.798
Content (mg/tablet) 70.85 71.03 69.15 68.38
RSD (%) (n = 5) 2.45 2.18 2.99 3.04
Sample C
ASA_KBr ASA_UATR
PCR+ PLS PCR+ PLS
RMS Error 0.2088A 0.1928A 0.2048A 0.1878A
Peak-to-Peak Error 0.9722A 0,8977A 0.9545A 0.8637A
Total M-Distance 0.925 0.884 0.910 0.978
Content (mg/tablet) 480.59 483.97 478.12 476.24
RSD (%) (n = 5) 1.99 2.17 2.87 3.01

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FT-IR Spectrophotometric analysis of acetylsalicylic acid and its pharmaceutical formulations
set of factors and each property is modeled by relating
the concentration values to those factors. In PLS1, the
spectra are modeled by a different set of factors for each
property and the concentration values are modeled by
the respective factors. Hence, PLS1 really contains n
separate calibrations, where n is the number of proper-
ties in the method.
We studied the possibility to use the Beer-Lambert
law for the quantitative determination of ASA in phar-
maceutical products at 1605.49 cm-1. The measurements,
carried out by the above mentioned conditions, provide
a typical calibration line which corresponds to:
A = - 1.2132 + 15.51 C(mg ASA)
with a regression coefficient, R, of 0.9886.
Figure 6 presents the calibration curve for ASA us-
ing the KBr disk method, while Tables 1 and 2 show the
results obtained by Beer’s law method and chemometric
approaches, where A, B and C represents the pharma-
ceuticals formulations manufactured by Bristol – Myers
Squibb Egypt, (sample A), from RAMEDA Co., Egypt
(sample B), and from Europharm, Romania (sample C),
respectively.
As can be seen in Table 1 and 2, the results are very
similar, and we suggest the use of the PCR+ method,
because of the smaller value of RSD (< 3.0%).
Conclusions
It is clear that FT-IR spectrometry is capable of
direct determination of ASA in several formulations.
With the commercial software involving chemometric
approaches, Beer-Lambert law, the method proposed is
simple, precise and not time-consuming compared to the
chromatographic methods that exist in literature. Quanti-
fication could be done in about 5-10 minutes, including
sample preparation and spectral acquisition.
Acknowledgements
One of the authors (AAB) wishes to thank the Admin-
istration of CROMATEC_PLUS SRL for the financial
support.
259
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