Simultaneous RP-HPLC Determination of Nimesulide and Paracetamol in Tablets
- Assay of nimesulide in pharmaceutical dosage formulation New Spectroscopic method for the estimation of Nimesulide in Pharmaceutical formulation: Her mejesty’s Stationary office. 113117-1854..
- Kumar Simultaneous RP-HPLC nimesulide and diclofenac sodium. 407..
- The controller of Publications. 554..
International Journal of PharmTech Research
CODEN( USA): IJPRIF ISSN : 0974-4304
Vol.1, No.3, pp 514-516 , July-Sept 2009
Simultaneous RP-HPLC Determination of Nimesulide and
Paracetamol in Tablets
Department of Quality Control, Nosch Laboratories Limited, Hyderabad-500072, India
Abstract : A simple, specific, accurate and precise reverse phase high pressure liquid chromatographic method has been
developed for the simultaneous determination of nimesulide and paracetamol from tablets by reverse phase C18 column
(Inertsil C18, 5m , 150 mm x 4.6 mm). The sample was analyzed using Acetonitrile: Methanol: Water in the ratio of 40:40:20,
(pH adjusted to 4.50 with orthophosphoric acid) as a mobile phase at a flow rate of 1.0 ml/min and detection at 276 nm. The
retention time for paractetamol and nimesulide was found to be 2.04 and 4.67 min respectively, and recoveries from tablet
were between 99 and 101 %. The method can be used for estimation of combination of these drugs in tablets.
Key words: Nimesulide, Paracetamol, RP-HPLC
Nimesulide is an anti-inflammatory drug. Chemically,
nimesulide is N-(4-nitro-2-phenoxyphenyl) methane
sulphonamide.It is approved for used in treatment of
thrombophlebitis and dental pain, inflammation. Some
HPLC1, 2 and spectrophotometric3, 4, methods have been
reported in the literature for its estimation. Paracetamol is
chemically N-(4-hydroxyphenyl) acetamide. It is a
centrally and peripherally acting non-opioid analgesic
and antipyretic. Many methods have been described in
the literature for the determination of paracetamol with
other drugs individually and in combination5-13. However
there is no RP-HPLC method reported for the
simultaneous estimation of these drugs in combined
A High Performance Liquid Chromatograph system, with
LC solutions data handling system (Shimadzu-LC2010)
with an auto sampler was used for the analysis. The data
was recorded using LC 2010 solutions software. The
purity determination performed on a stainless steel
column 150 mm long, 4.6 mm internal diameter filled
with Octadecyl silane chemically bonded to porous silica
particles of 5mm diameter (Inertsil C18, 5m , 150 mm x 4.6
mm, make: Shimadzu
chromatographic conditions are listed in Table 1.
Materials and Chemicals
ltd, Japan). Optimized
Pure samples of Nimesulide and Paracetamol were
obtained from Granules India ltd. For the estimation of
Nimesulide and Paracetamol in commercial formulations.
HPLC grade Orthophosphoric acid, acetonitrile and
methanol- procured from Merck, India. High pure water
was prepared by using Millipore Milli Q plus purification
Standard stock solution (1 mg/ml) of Nimesulide and
Paracetamol were prepared by dissolving 25 mg of drug
in 25 ml of acetonitrile, separately. The solutions were
suitably diluted with mobile phase to get mixed standard
solution containing 3 mg/ml of nimesulide and 15 mg/ml
Twenty tablets (Nimupain plus Cipla laboratories). Each
tablet was labeled contain 100 mg of Nimesulide and
Paracetamol 325 mg were weighed, and powder
equivalent to 25 mg of paracetamol was weighed
accurately and taken into 25 ml volumetric flask. The
drugs were extracted into acetonitrile; volume was
adjusted to 25 ml, vortexed and then filtered through 0.45
m membrane filter. From this solution, further dilutions
were made using mobile phase to get a final
concentration of 3 mg/ml of nimesulide and 15 mg/ml of
paracetamol. Twenty microliters of solution was injected
into HPLC system to obtain chromatogram for standard
drug solution (five replicates) and sample solution (five
paracetamol in the formulation were calculated by
comparing AUC of sample with that of standard.
of nimesulide and
Prasanna Reddy.Battu /Int.J. PharmTech Res.2009,1(3)515
Linearity and range of method was determined on
standard solution by analyzing 70 to 130 % of test
concentration, and the calibration curve was plotted using
AUC versus concentration of standard solution. Accuracy
of method was ascertained by recovery study by adding a
known amount of standard drug (±20% of test
concentration) to preanalysed sample and reanalyzing the
samples by proposed method. Precision was studied by
analyzing five replicates of sample solution. Specificity
was carried out by exposing the sample to different stress
conditions for 24 hours, such as acidic (0.1 N HCl, 1 ml
40OC), basic (0.1 N NaOH, 1 ml 40OC), heat (60OC), UV
light (260 nm, 40OC) and humidity (75 % RH, 40OC),
before analysis by proposed method. Ruggedness14 of
method was evaluated by performing the assay with
different analysis and on different days.
The chromatographic parameters were also validated by
system suitability studies (Table 2), which were carried
out on freshly prepared standard stock solutions. The
typical chromatogram obtained from the formulation is
presented in fig 1. The retention time for paracetamol and
nimesulide was found to be 2.04 and 4.67 min
respectively. Peaks were well resolved with resolution of
4.50 between the two drugs and were symmetrical in
shape with asymmetry factor less than 1.20. Linearity
was observed in the concentration range of 1.7-4.2 mg/ml
for nimesulide and 9-20 mg/ml for paracetamol, with the
correlation coefficient of 0.9996 for nimesulide and
0.9999 for paracetamol, respectively. Accuracy of the
method was ascertained by recovery study (n=3). The
concentration of standard spiked to the sample was 2.3-
mg/ml for nimesulide and 12-15
paracetamol. Recovery data from the study are reported
in table 3. The method was found to be accurate with
percent recoveries between 99 and 101 %. There was
good repeatability of proposed method with coefficient of
variance of 0.80% for nimesulide and 0.60% for
paracetamol. The results of specificity studies indicated
no interference from excipients, impurities, and
degradation products under various stress conditions and
assured that the peak response was due to a single
component only. Hence, the present method is cost-
effective, faster and can be used for the routine analysis
of these drugs from tablet formulations.
Table 1: Optimized Chromatographic conditions
Inertsil C18, 5m , 150 mm x 4.6 mm
Acetonitrile:methanoil:water (40:40:20) pH 4.5 (dil
UV at 276 nm
*Filtered through a 0.45m membrane filter (Millipore), degassed and sonicated
Table 2. System Suitability Parameters
Calibration range (mg/ml)
1.7 – 4.2
9 - 20
Table 3.Analysis of Formulation and Recovery studies.
Prasanna Reddy.Battu /Int.J. PharmTech Res.2009,1(3) 516
mg/tablet % label claim Amount added (mg/ml) % Recovery
Nimesulide 10099.98 99.98(0.82)
*mean (%RSD) of five observations,**mean (%RSD) of three determinations
Figure 1: Typical chromatogram of Nimesulide and Paracetamol
1.Zarapkar SS, Bhandari NP, Halkar UP.
Simultaneous determination of nimesulide and
chlorzoxazone in pharmaceutical dosage by RP-
HPLC. Indian Drugs 2000; 37:467.
Nagoji KE, Vijaysrinivas S, Kumar KM,
Mathivanan N, Kumar
nimesulide and diclofenac sodium. Indian J
Pharma Sci 2003; 65:407.
Rajput SJ, Randive G Assay of nimesulide in
pharmaceutical dosage formulation. Eastern
Pharmacist 1997; 475:113
Nagoji KE, Rao SS, Rao ME, Rao KV. New
Spectroscopic method for the estimation of
Nimesulide in Pharmaceutical
formulation. Eastern Pharmacist 1999; 496:117
British pharmacopoeia, Vol II, London: Her
mejesty’s Stationary office 1998, p. 1854.
Indian pharmacopoeis Vol II, New Delhi, The
controller of Publications. Govt of India, 1996,
SM, Rao ME.
7.Hasan, N.Y., Abdel-Elkawy, M., Elzeany, B.E.
and Wagieh, N.E., Farmaco., 2003, 58, 91.
EI-Saharty, Y.S., Refaat, M. and EI-Khateeb,
S.Z., Drug. Develop. Ind. Pharm., 2002, 28, 571.
Zawilla, N.H., Mohammad, M.A., EI-Kousy,
N.M. and EI-Moghazy Aly, S.M., J.Pharma.
Biomed. Anal., 2002, 27, 243.
10. Hinz, B., Auge, D., Rau, T., Rietbrock, S.,
Brune, K. And Werner, U., Biomed.
Chromatogr., 2003, 17, 268.
11. Liu, XQ., Chen, X.J., Zhao, L. H. And Peng,
J.H., Yao Xue Ba., 1997, 32, 546.
12. Lee, H.S., Jeong, C.K., Choi, S.J. Kim, S.B.,
Lee, M.H., Ko, G.I, and Sohn, D.H., J. Pharm.
Biomed. Anal., 2000, 23, 775.
13. EI-Kousy, N.M., J. Pharm. Biomed. Anal., 1999,
14. ICH, Validation of Analytical procedure:
Methodology (Q2B), International Conference
on Harmonization, IFPMA, Geneva, 1996