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

GPAT courses

Pharma courses

pharma courses

pharma courses



8. Limit of Quantification: It is defined as lowest concentration of analyte in a sample that can be determined with acceptable precision and accuracy and reliability by a given method under stated experimental conditions. LOQ is expressed as a concentration at a specified signal to noise ratio.

Limit of Quantification for Rabeprazole and Mosapride:The lowest concentration of the sample was prepared with respect to the base line noise and measured the signal to noise ratio. Limit of Quantificationis the lowest concentration of the substance that can be estimated quantitatively. It can be determined from linearity curve by applying the followingformula

Where S – slope of the calibration curve
σ – Residual standard deviation


 
9. Robustness: As part of the Robustness, deliberate change in the flow rate, mobile phase composition, temperature variation was made to evaluate the impact on the method. The standard and samples of Rabeprazole and Mosapride were injected by changing the conditions of chromatography. There was no significant change in the parameters like resolution, tailing factor, asymmetric factor, and plate count.The data are represented in table no. 11 and 12 and fig. no. 7, 8 and 9.

Table No. 11: It shows the system suitability results for Rabeprazole(Change in Flow Rate)

Flow 0.8 mL/min.

Std Area

Tailing factor

Flow 1.0 mL/min.

Std Area

Tailing factor

Flow 1.2 mL/min.

Std Area

Tailing factor

 

2588404

2588146

2588507

2588340

2588295

1.212666

1.231926

1.219733

1.218720

1.217223

 

2120053

2120059

2120201

2120054

2120451

1.214954

1.215568

1.207595

1.217034

1.214530

 

1730893

1730892

1730548

1730620

1730742

1.289723

1.284669

1.285484

1.284423

1.285398

Avg.

SD

% RSD

2588338

133.8219

0.01

1.220054

0.007166

0.57

Avg.

SD

% RSD

2120164

172.6146

0.01

1.213936

0.003

0.25

Avg.

SD

% RSD

1730739

156.3458

0.01

1.285939

0.002

0.16

Table No. 12: It shows the system suitability results for Mosapride(Change in Flow Rate)

Flow 0.8 mL/min.

Std Area

Tailing factor

Flow 1.0 mL/min.

Std Area

Tailing factor

Flow 1.2 mL/min.

Std Area

Tailing factor

 

1919212

1919607

1918031

1919556

1919620

1.144564

1.134991

1.130453

1.135498

1.134825

 

1440041

1440064

1440420

1440309

1440984

1.141374

1.136440

1.146321

1.147756

1.145364

 

1291932

1291600

1291369

1291294

1291498

1.286913

1.303066

1.313891

1.303122

1.303542

Avg.

SD

% RSD

1919205

677.3763

0.035

1.136066

0.005166

0.45

Avg.

SD

% RSD

1440364

382.3902

0.02

1.143451

0.00458

0.35

Avg.

SD

% RSD

1291539

249.3868

0.01

1.302107

0.009

0.69


Fig. no. 7: It shows typical chromatogram for robustness with flow rate
(for 0.8 mL/min flow)

Fig. no. 8: It shows typical chromatogram for robustness with flow rate
(for 1.0 mL/min flow)

Fig. no. 9: It shows typical chromatogram for robustness with flow rate
(for 1.2 mL/min flow)

RESULTS AND DISCUSSION
To optimize the mobile phase, various proportions of ammonium acetate buffer (pH 4.0) with methanol [HPLC Grade] were tested. The use of ammonium acetate buffer (pH 4.0) and methanol [HPLC Grade] in the ratio of 45:55 (v/v) resulted in peak with good shapes and resolution. A flow rate of 1.0 mL /min was found to be optimum in the 0.4-1.5 mL/min range resulting in short retention time, baseline stability and minimum noise.

By applying the proposed method, the retention times of rabeprazole and mosapride were observed at 2.946 and 4.186 minat 276 nm respectively. A typical chromatogram is represented in fig. no. 10.

Fig. No. 10 It shows typical chromatogram for Rabeprazole and Mosapride

Quantitative linearity was obeyed in the concentration ranges of 20-80 µg/mL for both rabeprazole and mosapride. The relevant regression equations were y = 53002x + 301.7 for rabeprazole (r2= 1) and y = 35995x + 707.7 for mosapride (r2= 1) (where y is the peak area ratio and x is the concentration of rabeprazole and mosapride (µg/mL)). The intra-day and inter-day drugs variations by the proposed method showed an RSD less than 2 %, indicating that the method is precise. The corresponding mean recoveries of the drugs were 99.98- 100.03 % for rabeprazole and 99.97 - 100.02 % for mosapride. This reveals that the method is quite accurate. The tailing factor (1.21 and 1.14 for rabeprazole and mosapride), USP plate count (11815.9 and 9435.372 for rabeprazole and mosapride); obtained were within the acceptance limits. The limits of detection for rabeprazole and mosaprideobtained by the proposed method were 0.01 and 0.035 µg/mL respectively, and limits of quantification for atorvastatin and ezetimibe obtainedby the proposed method were 0.032and 0.11 µg /mL respectively, which indicate the sensitivity of the method. The method tolerated minor variations in optimized chromatographic conditions indicating good robustness, which indicate the efficient performance of the column. 

No interfering peaks were found in the chromatograms indicating that the excipients used in tablet formulations did not interfere with the estimation of the drug by the proposed HPLC method.

CONCLUSION
The proposed HPLC method was found to be simple, precise, accurate and sensitive for the simultaneous determination of rabeprazole and mosapride. The method was validated as per ICH guidelines and all the parameters met within the acceptance criteria. Applicability of this method for simultaneous estimation of rabeprazole and mosapridefrom tablet dosage forms was confirmed. Hence, this  method  is  specific and  can  be  successfully  used  for  the  simultaneous estimation of rabeprazole and mosapridein bulk drug samples, pharmaceutical dosage forms. Hence, this method can be easily and conveniently adopted for routine quality control analysis of the above drugs.

ACKNOWLEDGEMENT: The authors greatly acknowledge M/s. Pharma Train, Hyderabad, Telagana, India for providing the gift sample of rabeprazole and mosapride.

REFERENCES
1. Carswel C. I. and Goa K L; Rabeprazole: an update of its use in acid related disorders; Drugs 2001; 61(15); 2327-2356.
2. Bhavesh Patel H., Madhabhai Patel M., Jignesh Patel R. and Bahnubhai Suhagia N; HPLC analysis for simultaneous determination of Rabeprazole and Domperidone in pharmaceutical formulation; J. Liq. Chrom. Rel. Tech. 2007; 30(3); 439-445.
3. Prasanna B. R. and Reddy M. S; Development and validation of RP-HPLC for the determination of Rabeprazole sodium in pharmaceutical formulations and human plasma; Asian J Res Chem; 2009; 2(1); 495-499.
4. Garcia CV, Paim C.S. and Steppe M; New liquid chromatographic method for determination of Rabeprazole sodium in coated tablets; J AOAC Int; 2004; 87(4); 842-846.
5. Rajesh S., Ganesh P. M., Subhash C. C; Development and validation of RP-HPLC method for the simultaneous determination of Rabeprazole sodium and Itopride hydrochloride in solid dosage form; E J Chem.; 2010; 7(3); 947-952.
6. Pillai S. and Singhvi I; Quantitative estimation of Itopride hydrochloride and Rabeprazole sodium from capsule formulation; Indian J Pharm Sci.; 2008; 70(5); 658-661.
7. Cassia Garcia V., Norma Nudelman S., Martin Steppe. and Elfrides Schapoval E.S;  Structural elucidation of Rabeprazole sodium photo degradation products; J. Pharm Biomed Ana; 2008; 46(1); 88-93.
8. Patel B. H., Suhagia B. N. and Patel M. M; High-performance liquid chromatography and thin-layer chromatography for the simultaneous quantitation  of Rabeprazole and Mosapride in pharmaceutical products; J Chrom Sci; 2008; 46(1); 4-10.
9. Shan Ren., Mi-Jin Park., Hongkee Sah. and Beom-Jin Lee; Effect of pharmaceutical excipients on aqueous stability of Rabeprazole sodium; Int J Pharm; 2008; 350(1-2); 197-204.
10. Osman A. O; Spectrofluorimetry, thin layer chromatography and column high-performance liquid chromatography determination of Rabeprazole sodium in the presence of its acidic and oxidized degradation products; J AOAC Int.; 2009; 92(5); 1373-1381.
11. El-Gindy A., El-Yazby F. and Maher M. M; Spectrophotometric and chromatographic determination of Rabeprazole in presence of its degradation products; J Pharm Biomed Anal. 2003; 31(2); 229-242.
12. Pattanayak P., Sharma R. and Chaturved S. C; Simultaneous spectrophotometric estimation of Rabeprazole sodium and Itopride HCl; Anal Lett; 2007; 40(12); 2288-2294.
13. Suganthi A., Sofiya J. and Ravi T. K; Simultaneous HPTLC determination of Rabeprazole and Itopride hydrochloride from their combined dosage form. Indian J Pharm Sci; 2008; 70(3); 366-8.
14. Shirkhedkar A. A. and Surana S. J; Application of stability-indicating RP-TLC densitometric determination of Rabeprazole sodium in bulk and pharmaceutical formulation; Eurasian J Anal Chem; 2009; 4(1); 165-170.
15. US Food and drug administration; Guidance for industry: Q2B validation of analytical procedures: methodology, Rockville; 1996.
16. US FDA; Guideline for industry: text on validation of analytical procedures: ICH Q2A; Rockville, MD; 1995.
17. International conference on harmonization (ICH), ICH quality guidelines: Good manufacturing practice guidance for active pharmaceutical ingredients Q7A (ICH), Geneva, Switzerland; 2001.
18. US Food and drug administration; Guidance document for industry; Analytical procedures and methods validation, FDA, Rockville, MD; 2000.
19. ICH, ICH Quality guidelines validation on analytical procedures: Methodology Q2B, ICH, Geneva, Switzerland; 1996.
20. Indian  Pharmacopeia  2007;  Volume  I; Published  by  The  Indian  Pharmacopoeia Commission; 477-478.

NOW YOU CAN ALSO PUBLISH YOUR ARTICLE ONLINE.

SUBMIT YOUR ARTICLE/PROJECT AT editor-in-chief@pharmatutor.org

Subscribe to Pharmatutor Alerts by Email

FIND OUT MORE ARTICLES AT OUR DATABASE


 

Pages

FIND MORE ARTICLES