METHOD DEVELOPMENT AND VALIDATION FOR ESTIMATION OF NAPROXEN IN BULK SAMPLES AS WELL AS IN TABLET DOSAGE FORMS BY USING RP-HPLC

 

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ABOUT AUTHORS
S. Ashutosh Kumar*, Manidipa Debnath,Vaddi Pavan Krishna Kumar

Department of Pharmaceutical Analysis and Quality Assurance,
A.K.R.G College of Pharmacy,
Nallajerla, West Godavari, A.P
* ashu.mpharm2007@gmail.com

ABSTRACT
Purpose:
A simple, precise, accurate, rapid and economical reverse phase high-pressure liquid chromatographic method has been developed as per ICH norms for the estimation of Naproxen from pharmaceutical formulation.
Methods: The method was carried out on a Kromosil-C18 ODS column (150 mm X 4.6 mm; 5 µ) with a mobile phase consists of ammonium acetate buffer (adjusted to pH 4.0 with 1 % Triethyl amine): methanol (40:60 v/v) and filtered through a 0.45 µ cellulose nitrate filters. The flow rate was maintained in isocratic mode at 1.0 mL/min. The detection was carried out at 210 nm. The run time was 7.0 min.
Results: The retention time was 3.063 min for Naproxen. The developed method was validated in terms of accuracy, precision, linearity, limit of detection, limit of quantification and solution stability.
Conclusion: The proposed method was adequate sensitive, reproducible, and specific for the determination of Naproxen in bulk as well as in tablet dosage forms.

REFERENCE ID: PHARMATUTOR-ART-2432

PharmaTutor (Print-ISSN: 2394 - 6679; e-ISSN: 2347 - 7881)

Volume 4, Issue 9

Received On: 17/03/2016; Accepted On: 19/04/2016; Published On: 01/09/2016

How to cite this article: Kumar SA, Debnath M, Kumar VPK; Method development and validation for estimation of Naproxen in bulk samples as well as in tablet dosage forms by using RP-HPLC; PharmaTutor; 2016; 4(9); 33-39

INTRODUCTION
Naproxen [(S)-6-methoxy-α-methyl-2-naphthaleneacetic acid], (Fig. 1) is a non-steroidal anti-inflammatory drug (NSAID) commonly used for the reduction of moderate to severe pain, fever, inflammation and stiffness. “It works by inhibiting both the COX-1 and COX-2 enzymes. Like other NSAIDs” [1-3]. Stability indicating simultaneous estimation of assay method for naproxen and Esomeprazole in pharmaceutical formulations by RP-HPLC is reported in literature [4]. Several chromatographic methods were reported for estimation of naproxen in raw materials, solid dosage forms mainly tablet and blood-plasma by RP-HPLC [5- 8]. Although literature survey reveals that various methods were reported for naproxen for single estimation and in combination with others drugs. However, the preparation of mobile phase was difficult and expensive; retention time was more; detection done at higher wavelength. Considering all these fact a successful attempt has been made to estimate naproxen by RP- HPLC with photo diode array detector.

Fig. 1: Chemical Structure of Naproxen

MATERIALS AND METHOD [9-10]
Chemicals and Reagents Used:
The following chemicals were used for the process: Water [HPLC Grade], Naproxen [working standards], Methanol [HPLC Grade], Ammonium acetate and Triethyl amine. All the chemicals were procured from Standard Solutions, Hyderabad, Andhra Pradesh.

0.45 µ membrane filters (Advanced Micro Devices Pvt. Ltd., Chandigarh, India) were used for filtration of various solvents and solutions intended for injection into the column.

Apparatus and Chromatographic Conditions: The equipment used was High Performance Liquid Chromatography Equipped with Auto Sampler and DAD or UV Detector. The column Kromosil-C18 ODS column (150 mm X 4.6 mm; 5 µ) was selected. The flow rate was monitored at 1.0 mL/min. The detection was carried out at 210 nm. The injection volume selected 20 µL, the temperature of the column oven was maintained at 25 °C, the detector used was Photo diode array and the run time was 7.0 min.

The ultra violet spectra of the drugs used for the investigation were taken on a Lab India UV 3000 spectrophotometer for finding out their λmax values.

Solubility of the compounds was enhanced by sonication on an ultra sonicator (Power Sonic 510, (Hwashin Technology).

All the weighings in the experiments were done with an Afcoset electronic balance. The HermLe microlitre centrifuge Z100 (model no 292 P01) was used for the centrifugation process and Remi equipments (model no- CM101DX) Cyclomixer was used.

Glassware: All the volumetric glassware used in the study was of Grade A quality Borosil.

Preparation of buffer solution [11]: The buffer solution was prepared by weighing accurately 3.85 gm of ammonium acetate and transferred in a 1000 mL beaker. Initially about 900 mL water [HPLC grade] was added for the dissolution of the buffer. Finally, the volume was made upto the mark with the diluent. The pH was adjusted to 4.0 with triethyl amine.

Preparation of mobile phase: The mobile phase was prepared by mixing a mixture of above buffer 400 mL (40 %) and 600 mL of methanol HPLC (60 %) and degas in ultrasonic water bath for 5 minutes. Then, the solution was filtered through a 0.45 µ filter under vacuum.

Preparation of standard solution of Naproxen: About 100 mg naproxen was weighed accurately and transferred into a 100 mL clean and dry volumetric flask. Initially, the drug was mixed with 7 mL of diluent. The solution was sonicated for 15 min for complete dissolution of the drug. The final volume was made up to the mark with the same solvent. From the above prepared solution, about 5 mL was pipetted out and transferred into a 100 mL clean and dry volumetric flask. Initially, the solution was mixed with 70 mL of diluent. The solution was sonicated for 15 min for complete dissolution of the drug. The final volume was made up to the mark with the same solvent to get a concentration of 50 µg/mL of Naproxen.

Preparation of sample solution of Naproxen: Twenty tablets were weighed accurately and a quantity of tablet powder equivalent to 100 mg of Naproxen was weighed and dissolved in the 70 mL mobile phase with the aid of ultra sonication for 20 min. The content was diluted with 100 mL mobile phase to furnish the preparation of stock solution. The stock solution was filtered through a 0.45 µm Nylon syringe filter and 5.0 mL of the filtrate was diluted into a 100.0 mL volumetric flask to get the desired concentration of 50.0 µg/mL of Naproxen.

System Suitability: The tailing factor for the peaks due to Naproxen in Standard solution should not be more than 2.0. The Theoretical plates for Naproxen peaks in Standard solution should not be less than 2000. The system suitability of the method was checked by injecting five different preparations of Naproxen. The parameters of system suitability were checked.

VALIDATION DEVELOPMENT [12-21]
1. System Suitability:
A Standard solution was prepared by using Naproxen working standards as per test method and was injected five times into the HPLC system. The system suitability parameters were evaluated from standard chromatograms by calculating the % RSD from five replicate injections for Naproxen sodium, retention times and peak areas. The data are represented in table no. 1.

Table no. 1: System Suitability data for Naproxen

Injection

RT

Peak Area

USP Plate count

USP Tailing

 

1

3.063

4437.5151

10168

1.106

2

3.064

4439.6279

10214

1.109

3

3.061

4437.5151

10200

1.110

4

3.059

4440.1612

10198

1.107

5

3.054

4446.1712

10210

1.108

Mean

3.0602

4440.198

10198

1.108

SD

0.003962

3.1749

-------

-------

% RSD

0.129479

0.0715

-------

-------

2. Specificity: Solutions of standard and sample are prepared as per the test method and are injected into chromatographic system. The chromatograms of standard and sample should be identical with near retention time. The specificity is represented in fig.no.2 and 3.

Fig. No. 2 A typical chromatogram for standard drug

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