Int. J. Drug Res. Tech. 2011, Vol.

1 (1), 1-7

International Journal of Drug Research and Technology

Available online at http://www.ijdrt.com/
Original Research Paper
RP-HPLC METHOD FOR THE ESTIMATION OF ACECLOFENAC IN TABLET
DOSAGE FORM
Jat R.K.1*, R. C. Chhipa 1, S. Sharma 2
1
Gyan Vihar School of Pharmacy,
SGVU Jagatpura, Jaipur-302025, India
2
Department of Pharmaceutical Science,
GJU, Hisar-125001, India
ABSTRACT
A simple, sensitive and precise RP-HPLC method was developed for the determination of aceclofenac in
tablet dosage form. The RP-HPLC separation was achieved on hypersil C18 column (250 mm, 4.6 mm,
5µm) using mobile phase water: acetonitrile (55:45 v/v) at flow rate of 1 ml/min at ambient temperature.
Quantification was achieved with photodiode array detection at 277 nm over the concentration range of 1-
10 µm/ml. The method was validated statistically and applied successfully for the determination of
aceclofenac.
Keywords: LC-MS method, HPLC method, Aceclofenac, Tablet dosage form, Nonselective
cyclooxygenase inhibitor, Anti-inflammatory agent.

INTRODUCTION Sun Pharmaceutical Laboratories Limited,

Jammu with purity of 99.987% tablets of three
Aceclofenac is nonselective cyclooxygenase
different companies (ACLOFEN-100 mg from
inhibitor that is anti-inflammatory agent used for
M/s Ind. Swift Pharma Ltd., HIFENAC-150 mg
inflammation, headache, gout, rheumatoid
from M/s Intas Pharma Ltd. and ACIFON-200
arthritis. Aceclofenac is chemically [({2-[(2,6-
mg from M/s Zenon Pharma Ltd) were procured
dichlorophenyl)amino]phenyl}acetyl)oxy]acetic
from the local pharmacy in the market figure3.
acid with an empirical formula C16H13Cl2NO4,
A standard stock solution of Aceclofenac (1
representing molecular weight of 354.18g/mol
mg/ml) was prepared by dissolving 25 mg of
figure2.1 Literature survey revealed LC-MS and
drug in 25 ml of acetonitrile. 9-12 Working
HPLC methods for estimation of Aceclofenac in
standard solution (100 µm/ml) was prepared
human plasma and pharmaceutical dosage
from stock solution by proper dilution with
forms.2-7 So it was thought of interest to develop
acetonitrile and water mixture. A Younglin (LC -
a simple, specific and sensitive RP-HPLC
2010TH –Liquid Chromatograph) equipped with
method for determination of aceclofenac.
PDA detector. Promesil C18 ((250 mm, 4.6 mm,
MATERIALS AND METHODS 5µm) column and LC software were used.13 The
mobile phase used was water: acetonitrile (55:45
All the reagents used were of HPLC grade and
v/v) which was filtered through nylon 0.45 µm
analytical grade. Reference standard of
aceclofenac was supplied as gift sample from

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 Jat R.K. et al. / International Journal of Drug Research and Technology 2011, Vol. 1 (1), 1-7

membrane filter and degassed by ultrasonication for 15 min (table 1).14,15

RESULTS AND DISCUSSION The system suitability tests for HPLC were
carried out on freshly prepared solution of
Linearity of the method was investigated by
Aceclofenac (10 µg/ml) and parameters were
serially diluting the working standard to give a
studied. The results were summarized in Table4.
concentration range of 1-10 µm/ml and 20 µl
The linear regression date showed a good linear
from this was injected. The flow rate was
relationship over the concentration range of 1-10
maintained at 1 ml/min. temperature of column
µg/ml as summarized in (table3). The limit of
was kept ambient and the effluent was monitored
detection (LOD) and the limit of quantification
at 277 nm. Calibration curve (figure4) was
(LOQ) of the drug were found by scanning the
constructed by plotting concentration against
solution of aceclofenac having different lower
peak area. The method was validated for
concentrations and the LOD and LOQ were
linearity, precision, accuracy, specificity, limit of
found to be 0.5 and 1 µg/ml indicates that
detection and limit of quantification as per ICH
method is sensitive (table3) The intraday and
guidelines.
interday precision were determined by analyzing
Assay of tablets of Aceclofenac were performed standard solution of aceclofenac at three
(table4). Twenty tablets of each company of different concentration levels (6, 8, 10 µg/ml).
strength 100 mg, 150 mg and 200mg were The % RSD for intraday and interday precision
weighed and ground to a fine powder. A quantity was found to be 0.257 – 0.712% and 0.480-
of tablet powder equivalent to 10 mg of 1.080% respectively which indicate that method
Aceclofenac was transferred to 10 ml volumetric is precise (table3) Repeatability of the method
flask, dissolved and diluted with acetonitrile and was studied by injecting 10 µg/ml solution of
water mixture to obtain 1 mg/ml. The solution aceclofenac for six times and peak area was
was sonicated for 15 minute and filtered through measured and % RSD was calculated which was
0.45 µm membrane filter. The solution was found to be 0.195 shows repeatability of the
further diluted to obtain concentration 10 µm/ml. method (table3) Accuracy of the method was
Peak area of the above prepared tablet solutions evaluated by standard addition method in which
of aceclofenac were measured by using above appropriate portion of stock solutions of
mentioned chromatographic conditions and the Aceclofenac were spiked into blank placebo
amount of aceclofenac were found from matrix to produce concentrations of 80 100 and
regression equation. 120% of theoretical concentration. The mean
To optimize the HPLC parameters, several recovery of spiked samples obtained was in
mobile phase compositions were tried. Various range of 99.06 to 101.03 reveals no interference
mobile phases having different ratios of of excipients and shows that method is accurate
methanol, water and acetonitrile were tried. Drug (table5). The proposed validated method was
was retained in mobile phase consisting of successfully applied to determine aceclofenac in
methanol : water (60 : 40 v/v) and methanol : tablet form. The results obtained for tablets of
acetonitrile (55 : 45 v/v). In methanol : water (60 aceclofenac were comparable with the
: 40 v/v) tailing in the peak was observed. Good corresponding labeled amounts (0.5 mg/tab)
peak symmetry and satisfactory retention time (table3) Robustness of the method was estimated
was obtained with mobile phase consisting of by changing the mobile phase composition
water : acetonitrile (55:45 v/v). Quantification (3±3), wavelength ±1 nm, injection volume
was achieved with PDA detection at 277 nm (20±2µl), column temperature (40±30) and RSD
based on peak area. The retention time of values for all these changes calculated were less
aceclofenac obtained was 6.60±0.132 (figure1). than 2 indicate that proposed method is robust.

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 Jat R.K. et al. / International Journal of Drug Research and Technology 2011, Vol. 1 (1), 1-7

The proposed RP-HPLC method was accurate, The proposed method shall prove equally
precise, sensitive and rapid. The method also can effective to analyze aceclofenac in the
be extended for the routine analysis of corresponding drug sample and may prove to be
aceclofenac in tablet dosage form. of great importance in pharmaceutical analysis.
CONCLUSION ACKNOWLEDGMENTS
It is thus concluded that the proposed method is Authors are grateful to Suresh Kalwania (Senior
new, simple, cost effective, accurate, safe, free Chemist) and M/s. Sun Pharma Lab, Jammu for
from pollution and precise and can be providing the gift samples of drugs. Authors are
successfully employed in the routine analysis of also thankful to Dr. R.K. Maheshwari for
this drug in pharmaceutical tablet dosage forms. valuable suggestions.

Table 1: HPLC instrumentation & chromatographic conditions for Aceclofenac

Parameters Description

Instrument A HPLC instrument (Younglin series) with Model Acme-9000

Column Promosil C-18, (250 mm, 4.6 mm, 5µm)
Mobile Phase Different mobile phase used for Trial 1 to 6
Flow Rate 1.0 mL/minute
Detection wavelength 277 nm
Injection Volume 10L
Run Time 10 minutes

Table 2: System Suitability Test Parameters

Parameters RP-HPLC method
Retention time, min 6.60±0.132
Tailing factor 1.048±0.274
Asymmetry factor 1.123±0.472
Theoretical plates 5859±0.774
Resolution 2.895±0.431

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Table 3: Regression characteristics and Validation Parameters

Sr. No. Parameter Value
1. λmax (nm) 277
2. Linearity range 1– 10
3. Correlation coefficient (r2) 0.999
4. Regression equation Y=0.99637X+49793
5. Intercept (a) .49793
6. Slope (b) 0.99637
7. Limit of detection (LOD μg/ml) 0.235
8. Limit of quantification(LOQ μg/ml) 0.869
9. Accuracy (%) 99.06-101.03
10. Repeatability (RSD, %, n=6) 0.195
11. Precision (RSD %), Interday (n=3) 0.438-1.080%
12. Intraday (n=30 0.257-0.712

Table 4: Results of analysis of commercial tablets of Aceclofenac

Tablet Label % Label claim % Coeff. of Standard
Formulation claim(mg) Estimated*(Mean ± S.D.) Variation error
I(ACLOFEN) 100 99.435 ± 1.243 1.365 0.514
II(HIFENAC) 150 99.754 ± 1.509 1.523 0.625
III (ACIFON) 200 99.246 ±1.427 1.305 0.613
*Average of six determinations
Table 5: Recovery studies of commercial tablets of Aceclofenac
Tablet Label Drug % Label claim % Coeff. of Standard
Formulation claim(mg) added(mg) estimated*(Mean ± S.D.) variation error

I(ACLOFEN) 100 50 99.316 ± 1.513 1.496 0.743

II(HIFENAC) 150 70 99.514 ± 1.397 1.432 0.574
III (ACIFON) 200 100 99.288 ±0.863 0.798 0.465
*Average of six determinations

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Jat R.K. et al. / International Journal of Drug Research and Technology 2011, Vol. 1 (1), 1-7

Figure1: Overlay chromatogram of standard Aceclofenac (10 µg/ml)

O
OCH2COOH
CH 2
Cl

NH

Cl

Figure2: Aceclofenac

Figure3: Infra Red Spectrum of Aceclofenac

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Concentration Linearity

9000000
8000000
7000000
6000000
y = 99637x + 49793
5000000
Area

R² = 0.999
4000000
3000000
2000000
1000000
0
0 1 2 3 4 5 6 7 8 9

Conc. Injected (ppm)

Figure4: Standard Curve of Aceclofenac

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