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Der Pharmacia Sinica, 2011, 2 (3): 164-171

ISSN: 0976-8688
CODEN (USA): PSHIBD

Development and validation of UV spectrophotometric methods for simultaneous

estimation of Paracetamol and Ibuprofen in pure and tablet dosage form
Rupali S. Joshi*, Nilima S. Pawar, Sameer S. Katiyar, Devendra B. Zope and Amol T. Shinde

Pharmaceutical Chemistry Department, P.D.V.V.P.F’s College of Pharmacy, Vilad – Ghat, Post

MIDC, Ahmednagar(M.S.) India
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ABSTRACT

Two simple, rapid, accurate, precise, and economic spectrophotometric methods for
simultaneous estimation of paracetamol and ibuprofen in pure and tablet dosage form have been
developed. Method I is based on solving simultaneous equation. Paracetamol and ibuprofen
show absorbance maximums at 256 and 222.4 nm respectively, so absorbance was measured at
the same wave lengths for the estimation of paracetamol and ibuprofen. Method II is based on
determination of Q-value. Absorbance is measured at 226.4 nm (Isoabsorptive point) and
222.4nm (λmax of ibuprofen). Both drugs obey the Beer Lambert's law in the concentration
range of 5-30 µg/mL. Methods are validated according to ICH guidelines and can be adopted
for the routine analysis of paracetamol and ibuprofen in pure and tablet dosage form.

Key words: Paracetamol, Ibuprofen, Simultaneous equation, Absorbance ratio, Validation.

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INTRODUCTION

Chemically, paracetamol is [N-(4-hydroxyphenyl)acetamide]. It is an analgesic antipyretic agent.

It is effective in treating mild to moderate pain such as headache, neuralgia and pain of musculo-
skeletal origin [1, 2]. The most recent methods for determination of paracetamol included
chromatographic [3-5], electrochemical [6-8] and spectrophotometric [9-11] techniques.

The 2-arylproprionic acid derivative, Ibuprofen [RS-2-(4-isobutyl-phenyl)propionic acid], is one

of the most potent orally active antipyretic, analgesic and nonsteroidal anti-inflammatory drug
(NSAID) used extensively in the treatment of acute and chronic pain, osteoarthritis, rheumatoid
arthritis and related conditions. Ibuprofen is characterized by a better tolerability compared with

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other NSAIDs. The techniques most recent used for determination of ibuprofen included
chromatographic [12-15], electrochemical [16,17] and spectrophotometric [18-23] methods.

Method validation is an important issue in pharmaceutical analysis. It confirms that the analytical
procedure employed for the analysis is suitable and reliable for its intended use. In present study,
all validation parameters for quantitative analysis of paracetamol and ibuprofen in tablets were
tested and data were evaluated according to their acceptance criteria.

The review of literature revealed that the combined dosage form has been estimated by
spectrophotometric method by using methanol as solvent in soft gelatin capsules [24] but no
method is yet reported in solvent 0.1N NaOH. Furthermore methanol being volatile in nature
creates problem in accuracy while 0.1 N NaOH being aqueous in nature serves for the accuracy.
Also methanol is more expensive than NaOH. Thus, this method is more accurate and cost
effective. This paper describes two simple, rapid, accurate, precise and economical methods for
simultaneous determination of paracetamol and ibuprofen in tablet dosage form.

MATERIALS AND METHODS

Instruments
UV-visible double beam spectrophotometer, Systronics model 2201 with spectral bandwidth of 1
nm, wavelength accuracy of ± 0.3 nm and a pair of 10 mm matched quartz cells was used and
Shimadzu AY220 balance was used for weighing.

Materials
Pure paracetamol was kindly gifted from Zest pharma, Indore and ibuprofen from Wanbury ltd.
Navi Mumbai. The commercially available tablets, Combiflam (Label claim: paracetamol- 325
mg, ibuprofen- 400 mg) was procured from local market. All the chemicals and reagents were of
analytical grade.

Selection of common solvent

After assessing the solubility of drugs in different solvents 0.1N NaOH has been selected as
common solvent for developing spectral characteristics.

Selection of wavelength
A representative overlain spectrum of Ibuprofen and Paracetamol in 0.1N NaOH is shown in Fig
1. The dilution was obtained to the concentration of 10 µg/ml for both paracetamol and ibuprofen
solution. Both the solutions were scanned in UV range (200-400nm) in 10 mm cell against
solvent blank. The study of spectrum revealed that paracetamol show a well defined λmax at 256
nm whereas ibuprofen shows at 222.4 nm. These two wavelengths were selected for
development of simultaneous equation. From the overlain spectrum, it is evident that
isoabsorptive point is at 226.4 nm for absorbance ratio method.

Preparation of standard stock solution and Study of Beer-Lambert’s Law

The standard stock solutions of paracetamol and ibuprofen were prepared by dissolving 0.025
gm of each drug in 0.1N NaOH and final volume was adjusted with same solvent in 100 mL of

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volumetric flask to get a solution containing 250 µg/mL of each drug. Aliquots of working stock
solutions of paracetamol and ibuprofen were diluted with 0.1N NaOH solution to get
concentration in range of 5-30 µg/ml for both the individual drug. The absorbances of resulting
solutions were measured at their respective max and isobestic point. A calibration curve as
concentration vs. absorbance (Fig-2) was constructed and by calculating absorptivity of both
drugs at respective wavelength; curve was constructed as concentration vs. absorptivity (Fig-3)
to study the Beer-Lambert’s Law and regression equation.

Method I (Simultaneous equation method)

If a sample contain two absorbing drug each of which absorbs at the λmax of the other, it may be
possible to determine both drugs by the technique of simultaneous equation.

Two wavelengths selected for the development of the simultaneous equations are 256 nm and
222.4 nm. The absorptivity values determined for paracetamol are 0.0643 (ax1), 0.0242 (ax2)
and for ibuprofen are 0.0055 (ay1), 0.0351 (ay2) at 256 nm and 222.4 nm respectively. These
values are means of six estimations. The absorbances and absorptivity at these wavelengths were
substituted in equation 1 and 2 to obtain the concentration of both drugs.

------------------------------Eqn.1
-------------------------------Eqn.2

Where Cparacetamol and Cibuprofen are concentration of paracetamol and ibuprofen

respectively in µg/mL. A1 and A2 are the absorbance of the mixture at 256 nm and 222.4 nm
respectively.

Method II (Absorbance ratio method)

Absorbance ratio method of analysis is based on the absorbance at two selected wavelengths, one
of which is an isoabsorptive point and the other being the wavelength of maximum absorption of
one drug. From overlain spectra (Fig. 1), 222.4 nm (λmax of ibuprofen) and 226.4 nm
(isoabsorptive point) are selected for the formation of Q absorbance equation (Eqn. 3 and 4). The
absorptivity values determined for paracetamol are 0.0242 (ax1), 0.0294 (ax2) and for ibuprofen
are 0.0351 (ay1), 0.0253 (ay2) at 222.4 nm and 226.4 nm respectively. These values are means
of six estimations. The absorbances and absorptivity at these wavelengths were substituted in
equation 3 and 4 to obtain the concentration of drugs.

-------------------------------Eqn.3
-------------------------------Eqn.4

Where Cparacetamol and Cibuprofen are concentration of paracetamol and ibuprofen

respectively in µg/mL. A1 and A2 were the absorbance of the sample at 222.4 nm and 226.4 nm
respectively.

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Analysis of the tablet formulations
Twenty tablets of marketed formulation were accurately weighed and powdered. A quantity of
powder equivalent to 50 mg of paracetamol was transferred to 100 mL volumetric flask and
dissolved in 0.1N NaOH and final volume was made up with 0.1N NaOH. The sample solution
was then filtered through Whatman filter paper No.41. From the above solution 10 mL of
solution was taken and diluted to 50 mL with 0.1N NaOH to get a solution containing 100
µg/mL of paracetamol and corresponding concentration of ibuprofen. From above, 0.65 mL of
solution was diluted with same solvent in 10 mL volumetric flask to get final concentration of
paracetamol 6.5µg/mL and ibuprofen 8µg/mL. Analysis procedure was repeated six times with
tablet formulation. The results of tablet analysis are reported in Table 2.

Validation of the developed methods

Linearity
For each drug, appropriate dilutions of standard stock solutions were assayed as per the
developed methods. For method I and II, the Beer- Lambert’s concentration range was found to
be 5-30 µg/mL for both drugs. The linearity data for both methods are presented in Table 1.

Accuracy
To check the accuracy of the proposed method, recovery studies were carried out 80, 100 and
120% of the test concentration as per ICH guidelines. The recovery study was performed three
times at each level. The result of the recovery studies are reported in table 3.

Precision:
Repeatability
To check the degree of repeatability of the methods, suitable statistical evaluation was carried
out. Repeatability was performed for six times with tablets formulation. The standard deviation,
coefficient of variation and standard error was calculated. The results of statistical evaluation are
given in Table 2.

Intermediate Precision (Interday and Intraday precision)

The interday and intraday precision was determined by assay of the sample solution on the same
day and on different days at different time intervals respectively. The results of the same are
presented in Table 4.

RESULT AND DISCUSSION

Linearity range for paracetamol and ibuprofen are 5-30 µg/mL at respective selected
wavelengths. The coefficient of correlation for paracetamol at 256 nm and for ibuprofen at 222.4
nm is 1 and 0.999 respectively. Both drugs shows good regression values at their respective
wavelengths and the results of recovery study reveals that any small change in the drug
concentration in the solution could be accurately determined by the proposed methods.

Percentage estimation of paracetamol and ibuprofen from tablet dosage form by method I is
98.98 and 99.50 and by method II is 98.69 and 99.68 respectively with standard deviation <2
(Table 2).

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Table No. 1: Optical parameters and regression characteristic
Paracetamol at Ibuprofen at
Paracetamol Ibuprofen
Parameters iso-absorptive iso-absorptive
at λmax at λmax
point point
Beer’s Law range 5-30 µg/mL 5-30 µg/mL 5-30 µg/mL 5-30 µg/mL
Regression Y = 0.064x Y = 0.034x Y = 0.029x Y = 0.0254x
Equation (Y) + 0.001 + 0.003 + 0.004 + 0.0002
Slope (m) 0.064 0.034 0.029 0.0254
Intercept (c) 0.001 0.003 0.004 0.0002
Correlation coefficient 1 0.999 0.999 0.9997

Table No. 2: Analysis of Tablet Formulation

Absorbance Reading at selected Conc. Acquired % Found (µg /ml)
wavelength Method I and II Method I Method II
256nm 222.4nm 226.4nm PAR IBU PAR IBU PAR IBU
0.456 0.432 0.388 6.5 8 98.54 98.31 98.50 98.30
0.455 0.433 0.389 6.5 8 98.21 98.86 98.81 98.49
0.457 0.438 0.392 6.5 8 98.53 100.24 98.05 100.69
0.464 0.441 0.395 6.5 8 100.18 100.56 99.12 101.16
0.457 0.439 0.393 6.5 8 98.49 100.81 99.44 100.83
0.462 0.434 0.390 6.5 8 99.95 98.23 99.22 98.62
Average 98.98 99.50 98.69 99.68
S.D. 0.8496 1.1684 0.4446 1.3399
COV 0.8584 1.1742 0.4505 1.3442
SE 0.3469 0.4770 0.1815 0.5470
PAR: Paracetamol, IBU: Ibuprofen, S.D: Standard Deviation, COV: Coefficient of variation, S.E: Standard Error.

Table No. 3: Result of recovery study

Method I Method II
PAR IBU PAR IBU
Amount Taken
6.5 8.0 6.5 8.0
(µg/ml)
% 80 100 120 80 100 120 80 100 120 80 100 120
Amount
µg/m
Added 5.2 6.5 7.8 6.4 8.0 9.6 5.2 6.5 7.8 6.4 8.0 9.6
l
99.89 99.00 100.02 98.98 98.50 99.50 99.50 98.95 100.05 99.60 98.60 99.00
% Recovery#
±0.82 ±1.14 ±0.78 ±0.64 ±1.30 ±0.88 ±0.97 ±1.64 ±1.28 ±0.59 ±1.35 ±0.71
Mean Recovery
99.63±0.91 98.99±0.94 99.5±1.29 99.06±0.88
±S.D.
PAR: Paracetamol, IBU: Ibuprofen, S.D: Standard Deviation,

Table No. 4: Validation parameters

Precision (%COV)
Method Drug Intraday Interday*
n=3 First day Second day Third day
PAR 0.5621 0.9204 0.7526 0.4425
I
IBU 0.9143 0.7562 0.9224 0.7666
PAR 0.7025 0.9546 0.6042 0.5480
II
IBU 0.6924 0.5574 0.6524 0.5669
PAR: Paracetamol, IBU: Ibuprofen, COV: Coefficient of variation, *Average of six determination.

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Fig.1 Overlain spectra of paracetamol and ibuprofen

Fig. 2. Study of Beer’s-lamberts law

The validity and reliability of proposed methods are assessed by recovery studies. Sample
recovery for both the methods is in good agreement with their respective label claims, which
suggest non interference of formulation additives in estimation (Table 3).

Precision is determined by studying the repeatability and intermediate precision. Repeatability

result indicates the precision under the same operating conditions over a short interval of time
and interassay precision. The standard deviation, coefficient of variance and standard error are
calculated for paracetamol and ibuprofen. The results are mentioned in Table 2. Intermediate
precision study expresses within laboratory variation in different days. In both intra and inter day
precision study for both the methods % COV are not more than 2.0% indicates good repeatability
and intermediate precision (Table 4).
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Fig.3 Response ratio curve A-Paracetamol at 256nm, B-Ibuprofen at 222.4nm, C-Paracetamol-226.4nm and
D-Ibuprofen at 226.4nm

CONCLUSION

The proposed spectrophotometric methods are simple, rapid, accurate, precise, and economic and
validated in terms of linearity, accuracy, precision, specificity and reproducibility. These two
methods can be successfully used for simultaneous estimation of paracetamol and ibuprofen in
pure and tablet dosage form.

Acknowledgement:
The authors are greatly thankful to P.D.V.V.P.F’s College of Pharmacy, Ahmednagar, India for
providing access to facilities and necessary infrastructure to carry out research work. We are also
thankful to for providing us the free gift sample of Ibuprofen by Wanbury ltd. Navi Mumbai and
Paracetamol by Zest pharma, Indore which were required for our research work.

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