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RP-HPLC METHOD FOR SIMULTANEOUS DETERMINATION OF IBUPROFEN AND FAMOTIDINE IN PHARMACEUTICAL DOSAGE FORM

L. Peikova*, M. Georgieva, B. Tsvetkova

Medical University Sofia, Faculty of Pharmacy, Department of Pharmaceutical chemistry,D2 Dunav str., 1000 Sofia, Bulgaria

Abstract: In this study, reversed phase high performance liquid chromatographic method have been de-veloped and validated for the simultaneous determination of ibuprofen and famotidine in combined phar-maceutical formulation. Separation was achieved with a C8 (250 mm x 4.6 mm, 5 m) column, ambient temperature with isocratic mode with mobile phase containing acetonitrile and 0.5 M potassium dihydrogen phosphate buffer pH 2.2 adjusted with ortho-phosphoric acid (25:75). UV detection was performed at 280 nm. The flow rate was 1.2 ml/min. The retention times of ibuprofen and famotidine were found to be 3.19 min and 8.37 min, respectively. The responses were linear (R2 > 0.9999) in the range of 20 160 g/ml for ibuprofen and 0.68 5.4 g/ml for famotidine. The % recovery for ibuprofen and famotidine was 99.75 and 99.24, respectively. No chromatographic interference from the tablet excipients was found. The results of the studies showed that the proposed RP-HPLC method is simple, rapid, precise and accurate, which can be applied for the routine analysis of ibuprofen and famotidine in tablet dosage forms.

Key words: liquid chromatography, validation, ibuprofen, famotidine, tablet dosage form, quality control

Original Articles

Introduction The 2-arylproprionic acid derivative, ibu-

profen, (RS)2-(4isobutyl phenyl) propionic acid is a nonsteroidal anti inflammatory drug (NSAID) which is used in reducing inflamma-tion and pain associated with many diseases like rheumatoid arthritis, osteoarthritis and related conditions. Ibuprofen is characterized by a better tolerability compared with other NSAIDs.

Famotidine is chemically 3-([2-(diamino-methyleneamino) thiazol- 4-yl] methylthio)-N-sulfamoylpropanimidamide. It is commonly ap-plied in the treatment of peptic ulcer disease and gastro esophageal reflux disease. Famotidine is histamine H2-receptor antagonist which blocks the action of histamine on stomach cells and re-duces acid production [1, 2].

A fixed dose combined dosage form of ibu-profen and famotidine is indicated for the relief of signs and symptoms of rheumatoid arthritis and osteoarthritis and to decrease the risk of de-veloping upper gastrointestinal ulcers. Ibupro-fen is helpful in relieving the pain and inflam-mation associated with arthritis and famotidine reduces the risk of gastric ulcers which is a side effect in chronic usage of ibuprofen [3]. The combination of these two drugs is not official in any pharmacopoeia.

Extensive literature survey revealed determi-nation of famotidine in dosage form by HPLC [4-10] and UV-spectrophotometry [11]. Analy-sis of ibuprofen in both pharmaceutical dosage forms and blood plasma has been reported by UV-spectrophotometry [12], capillary zone elec-trophoresis [13] and HPLC [14, 15]. Simultane-ous determination of ibuprofen and famotidine in combination includes liquid chromatographic techniques [16-18], UV- spectrophotometry [19-21] and thin layer chromatography [22].

The aim of the present study is to develop an analytical method for simultaneously determina-tion of ibuprofen and famotidine in a tablet for-mulation containing 400 mg ibuprofen and 13.3 mg famotidine using isocratic reversed phased liquid chromatographic procedure.

Materials and methodsReagents and chemicalsWorking standards of ibuprofen RS (Purity 100.1)

and famotidine RS (purity 99.92) were provided by (Sigma-Aldrich). The formulated tablets (label claim: ibuprofen 400 mg and famotidine 13.3 mg) were used for analysis. LC-grade acetonitrile and ortho-phosphoric acid were procured from Merck (Germany). All other chemical reagents were of analytical grade.

4 L. Peikova, M. Georgieva, B. TsvetkovaPHARMACIA, vol. 61, No. 2/2014

Instrumentation and chromatographic conditionsChromatographic separation was performed on

a modular HPLC system LC-10AShimadzu (Ja-pan) comprising a LC-10A pump, solvent degas-ser DGU-3A, Rheodyne injector with 20 l loop, column oven CTO-10A, SPD-M10A UV detector with fixed wavelength and communication bus module CBM-10A. Separation was achieved iso-cratically with a LiChrosorb C8, 250 mm x 4.6 mm, 5 m column eluted with a mixture of acetonitrile and 0.5 M potassium dihydrogen phosphate buffer pH 2.5 adjusted with ortho-phosphoric acid (25:75 v/v) as the mobile phase at flow rate of 1.2 ml/min. The mobile phase was filtered through a 0.45 m membrane filter and degassed. Detection was carried out by absorbance at 280 nm. The analysis was carried out at ambient column temperature and injection volume was 20 l.

Preparation of reference solutionsFor ibuprofen: 16 mg of ibuprofen was accu-

rately weighed and transferred to a 50 ml volumet-ric flask and volume was made up to the mark with methanol (Stock solution A - 320 g/ml).

For famotidine: 27 mg of famotidine was accu-rately weighed and transferred to a 100 ml volumet-ric flask and volume was made up to 100 ml with methanol (Stock solution B - 270 g/ml). From Stock solution B, 5.0 ml was taken into a 50 ml volumetric flask and volume was made up to 50 ml with metha-nol (Stock solution C - 27 g/ml).

For mixed standard: The reference solution was prepared by diluting 5.0 ml of stock solution A and 2.0 ml of stock solution C with methanol into a 20.0 ml volumetric flask (80 g/ml ibuprofen and 2.7 g/ml famotidine).

Sample preparationTablets working solution was prepared as fol-

lows: an accurately weighed quantity of powdered tablet sample containing an equivalent of 400 mg ibuprofen and 13.3 mg famotidine were transferred into 100 ml volumetric flask, then 70 ml methanol was added. The mixture was sonicated for ten min-utes and the volume was made up using methanol, then the sample was filtered. 2.0 ml of the filtrate was diluted with methanol into a 100.0 ml volu-metric flask to give a test solution containing 80 g/ml ibuprofen and 2.7 g/ml famotidine.

Results and discussionMethod validationThe proposed method was validated with respect

to specificity, linearity, precision, accuracy, limit of quantitation (LOQ) and limit of detection (LOD).

SpecificityThe specificity of the method was determined by

checking the interference of the components against placebo. No interference was observed for any of the excipients of both drugs. The Fig. 1 showed typical chromatogram obtained from analysis of standard so-lution using the proposed method. The retention time observed 3.19 min for ibuprofen and 8.37 min for famotidine permits a rapid assay, which is important for routine analysis.

Calibration and linearityTo establish the linearity of analytical method, a

series of dilution in the range from 20-160 g/ml for ibuprofen and 0.68-5.4 g/ml for famotidine were prepared. All the solutions were filtered through 0.22 m membrane filter prior to use and injected in chrommatograph. A calibration curves were plot-ted between the mean peak areas vs. respective con-centrations. The corresponding linear regression equation was y=10784x-1320.2 with square of cor-relation coefficient R2 of 0.9999 for ibuprofen and y=18540x-1264.4 with square of correlation coeffi-cient R2 of 0.9999 for famotidine, respectively.

Limit of quantitation and limit of detectionThe limit of quantitation and limit of detection

were calculated from the standard deviations and slopes of the responses using a signal-to-noise ratio. The LOQs for ibuprofen and famotidine were found to be 2 g/ml and 0.8 g/ml, while the LODs were 0.2 g/ml and 0.1 g/ml, respectively.

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Results and discussion

Method validation

The proposed method was validated with respect to specificity, linearity, precision,

accuracy, limit of quantitation (LOQ) and limit of detection (LOD).

Specificity

The specificity of the method was determined by checking the interference of the

components against placebo. No interference was observed for any of the excipients of

both drugs. The Fig. 1 showed typical chromatogram obtained from analysis of standard

solution using the proposed method. The retention time observed 3.19 min for

ibuprofen and 8.37 min for famotidine permits a rapid assay, which is important for

routine analysis.

0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 min

0.00

0.25

0.50

0.75

1.00

1.25

1.50

1.75

2.00

2.25

2.50

2.75

3.00

mV(x10)Detector A:280nm

3.19

5/98

7885

8.36

7/88

3900

Figure 1. Chromatogram obtained from analysis of standard solution

Calibration and linearity

To establish the linearity of analytical method, a series of dilution in the range from 20-

160 g/ml for ibuprofen and 0.68-5.4 g/ml for famotidine were prepared. All the

solutions were filtered through 0.22 m membrane filter prior to use and injected in

Fig 1. Chromatogram obtained from analysis of standard solution

RP-HPLC method for simultaneous determination of Ibuprofen ... PHARMACIA, vol. 61, No. 2/2014 5

AccuracyRecovery studies were performed to validate the

accuracy of developed method. To the preanalysed sample solution, a definite concentration of standard drug was added and then its recovery was analysed. The percent recovery for ibuprofen was found to be 99.75 % and for famotidine it was 99.24 %. Results presented in Table 1 indicated good accuracy and showed no interference from tablet excipients.

PrecisionThe precision of the method was evaluated by

performing six independent determinations of the test sample preparation and calculating RSD (%). The RSD values measured during assessment of pre-cision was

6 L. Peikova, M. Georgieva, B. TsvetkovaPHARMACIA, vol. 61, No. 2/2014

Corresponding authorL. PeikovaMobile: +359 (0)898 454851;Tel: +3599236538 ; Fax: +3599879874;E-mail address: lily_peikova@yahoo.com

12. M i t i c SS, Miletic GZ, Pavlovic AN, Arsic BB, Zivanovic VV. Quantitative analysis of Ibuprofen in pharmaceuticals and human con-trol serum using kinetic spectrophotometry. Journal of Serbian Chemical Society 2008; 73 (10): 879-890.

13. C h e n H, Huang D, Chen Q. Li H., Rapid de-termination of active constituents in compound Ibuprofen tablets by capillary zone electro-phoresis. Chinese Journal of Chromatography 1998; 16: 289-292.

14. H a i k a l a VE, Heimonen IK, Vuorela HJ. Determination of Ibuprofen in ointments by re-versed phase liquid chromatography. Journal of Pharmaceutical Sciences 1991; 80: 456-458.

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Journal of Pharma and Bio Sciences 2013; (3): 655-662.

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