AN ISOCRATIC METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS ESTIMATION OF RABEPRAZOLE AND MOSAPRIDE IN TABLET DOSAGE FORMS BY USING RP-HPLC

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ABOUT AUTHORS
S. Ashutosh Kumar*, Manidipa Debnath, Venugopal Padala,
Department of Pharmaceutical Analysis and Quality Assurance,
A.K.R.G College of Pharmacy, Nallajerla, West Godavari, A.P
*ashu.mpharm2007@gmail.com

ABSTRACT
Objective:
The present work was undertaken with the aim to develop and validate a rapid and consistent RP-HPLC method in which the peaks will be appear with short period of time as per ICH Guidelines.
Method: The HPLC separation was achieved on an Inertsil-C18 ODS column (250 X 4.6 mm; 5 µ) column in an Isocratic Mode. The mobile phase composed of Methanol [HPLC Grade] (55 %) and Buffer (45 %) [pH 4.0 adjusted with triethylamine]. The flow rate was monitored at 1.0 mL/min. The wavelength was selected for the detection was 276 nm.
Results:The retention times found for rabeprazole and mosapride was 2.946 and 4.186 min respectively. The % recovery was 99.98- 100.03 for rabeprazole and 99.97 - 100.02 for mosapride. The linearity was established in the range of 20-80 µg/mL for both rabeprazole and mosapride. The LOD for rabeprazole and mosapride were 0.01 and 0.035 µg/mL respectively. The LOQ for rabeprazole and mosapride were 0.032and 0.11 µg/mL respectively.
Conclusion:The proposed method was adequate sensitive, reproducible, and specific for the determination of rabeprazole and mosapride in bulk as well as in tablet dosage forms.

REFERENCE ID: PHARMATUTOR-ART-2422

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

Volume 4, Issue 7

Received On: 27/02/2016; Accepted On: 08/03/2016; Published On: 01/07/2016

How to cite this article: Kumar SA, Debnath M, Padala V; An Iso cratic Method Development and Validation for simultaneous estimation of Rabeprazole and Mosapride in Tablet Dosage Forms by using RP-HPLC; PharmaTutor; 2016; 4(7); 41-51

INTRODUCTION
Rabeprazole sodium is chemically (RS)-2-[(4-(3-methoxy propaxy)-3-methylpyridin-2-yl] methyl sulphonyl)-1H-benzo (d) imidazole (fig. 1). Rabeprazole sodium[1] is an antiulcer drug in the class of proton pump inhibitors. As anti ulcer drug, it is used in short-term treatment in healing and symptomatic relief of duodenal ulcers and erosive or ulcerative gastro esophageal reflux disease (GORD); maintaining healing and reducing relapse rates of heartburn symptoms in patients with GORD; treatment of daytime and nighttime heartburn and other symptoms associated with GORD; long-term treatment of pathological hypersecretory conditions, including Zollinger-Ellison syndrome and in combination with Amoxicillin and Clarithromycin to eradicate H. pylori. Mosapride citrate is chemically (RS)-4-amino-5-chloro-2-ethoxy-N-[(4-(4-fluorobenzyl) morpholin-2-yl) methyl] bezamide citrate (fig. 2). Mosapride is a gastro pro-kinetic agent that acts as a selective 5HT4 agonist. The major active metabolite of Mosapride is known as M1, additionally acts as a 5HT3 antagonist. In addition to its prokinetic properties, Mosapride also exerts anti-inflammatory effects on GIT which may contribute to some of its therapeutic effects. Mosapride also promotes neurogenesis in the gastrointestinal tract which may prove useful in certain bowel disorders. The neurogenesis is due to Mosapride’s effect on 5-HT4 receptor where it acts as an agonist. The drug analysis plays an important role in the development of drugs, manufacturing and therapeutic use. Pharmaceutical industries rely upon quantitative chemical analysis to ensure that the raw material used and the final product obtained meets the required specification.

Fig. no. 1 It shows the chemical structure of Rabeprazole

Fig. no. 2 It shows the chemical structure of Mosapride

The literature review indicates there are several analytical methods have been reported for estimation of these drugs as individual or in combination with other drugs, and also several analytical methods for the determination of simultaneous estimation of Rabeprazole sodium and Mosapride citrate by RP-HPLC[2-9], Spectrofluorimetry, thin layer chromatography and column high-performance liquid chromatography[10], Spectrophotometric and chromatographic[11-12], HPTLC[13] and TLC[14] in dosage formulation and/or in presence of its degraded products. Some of the reported RP-HPLC methods were not economical in terms of mobile phase composition, column dimensions and run times. Hence there is need for the development of newer method for estimation of Rabeprazole sodium and Mosapride citrate present in tablet to overcome above discussed hurdles. In addition, RP-HPLC method for the simultaneous estimation of Rabeprazole sodium and Mosapride citrate in pharmaceutical dosage form are very scanty. Hence the main objective of this study is to develop a RPHPLC method for estimation of Rabeprazole sodium and Mosapride citrate and validate the developed method according to ICH guidelines[15-19] by using various parameters.

MATERIALS AND METHODS
Chemicals and Reagents Used: The following chemicals were used for the processwater [HPLC Grade], rabeprazole and mosapride [working standards] gift samples collected from Pharma Train Lab., Hyderabad, Telagana., methanol [HPLC Grade], ammonium acetate and triethylamine. 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 Inertsil-C18 ODS column (250 X 4.6 mm; 5 µ) was selected. The flow rate was monitored at 1.0 mL/min. The detection was carried out at 276 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 10.0 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 Phosphate buffer[20]:The buffer solution was prepared by weighing accurately 3.85 gm of ammonium acetate and transferred to a clean and dry 1000 mL volumetric flask. Initially, about 900 mL of water [HPLC grade] was added. The final volume was made upto the mark with water. Then the pH was adjusted to 4.0 with triethylamine.

Preparation of mobile phase: The mobile phase was prepared by mixing a mixture of above buffer 450 mL (45 %) and 550 mL of methanol HPLC (55 %) and degas in ultrasonic water bath for 5 minutes. Then, the resultant solution was filtered through a 0.45 µ filter under vacuum.

Preparation of standard solution of Rabeprazole and Mosapride: About 10 mg rabeprazole was weighed accurately and transferred into a 10 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. Similarly, about 10 mg mosapride was weighed accurately and transferred into a 10 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 to get a concentration of 1000 µg/mL.

From the above prepared stock solutions 0.4 mL of rabeprazole and mosapride were pipetted out into a 10 mL clean and dry volumetric flask and it was diluted up to the mark with diluent. This mixed stock solution contains 40.0 µg/mL of rabeprazole and 40.0 µg/mL of mosapride.

Preparation of sample solution of Rabeprazole and Mosapride: Twenty tablets were weighed accurately and a quantity of tablet powder equivalent to 20 mg of rabeprazole and 20 mg of mosapride were 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 10.0 mL of the filtrate was diluted into a 50.0 mL volumetric flask to get the desired concentration of 40.0 µg/mL of rabeprazole and 40.0 µg/mL of mosapride.

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

VALIDATION DEVELOPMENT[15-19]
1. System Suitability:A Standard solution was prepared by using rabeprazole and mosapride 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 rabeprazole and mosapride, retention times and peak areas. The data are represented in table no. 1 and 2.

Acceptance Criteria: The % RSD for the retention times of principal peak from 5 replicate injections of each Standard solution should be not more than 2.0 %. The number of theoretical plates (N) for the Sumatriptan succinate and Naproxen sodium peaksis NLT 3000. The Tailing factor (T) for the Sumatriptan succinate and Naproxen sodium peaks is NMT 2.0.

Table no. 1: It shows the system suitability data for Rabeprazole

Injection

RT

Peak Area

USP Plate count

USP Tailing

1

2

3

4

5

Mean

SD

% RSD

2.951

2.950

2.948

2.949

2.949

2.947

0.003701

0.125589

2120053

2120059

2120201

2120054

2120451

2120164

172.6146

0.01

11898.457087

11844.975123

11857.288976

11809.408109

11669.365498

11815.9

-------

-------

1.214954

1.215568

1.207595

1.217034

1.214530

1.213936

-------

-------

Table no. 2: It shows the system suitability data for Mosapride

Injection

RT

Peak Area

USP Plate count

USP Tailing

1

2

3

4

5

Mean

SD

% RSD

4.195

4.193

4.189

4.190

4.188

4.191

0.002828

0.067488

1440041

1440064

1440420

1440309

1440984

1440364

382.3902

0.03

9559.400562

9468.102886

9470.850282

9425.185779

9253.320313

9435.372

-------

-------

1.141374

1.136440

1.146321

1.147756

1.145364

1.143451

-------

-------

2. Specificity: Solutions of standard and sample were prepared as per the test method are injected into chromatographic system. The chromatograms of standard and sample should be identical with near retention time. The specificity for method is represented in fig.no.3 and 4.

Fig. No. 3: It shows a typical chromatogram for standard drugs

Fig. No. 4: It shows a typical chromatogram for sample drugs

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