RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS QUANTITATIVE ESTIMATION OF MEBEVERINE HYDROCHLORIDE AND CHLORDIAZEPOXIDE IN CAPSULES
Objective: To develop an accurate, precise and linear Reverse Phase High Performance Liquid Chromatographic (RP-HPLC) method for simultaneous quantitative estimation of Mebeverine hydrochloride and Chlordiazepoxide in MEVA C capsules and validate as per ICH guidelines.
Methods: The optimized method uses a reverse phase column, Enable Make C18G (250 X 4.6 mm; 5Î¼), a mobile phase of triethylammonium phosphate buffer (pH 2.5): acetonitrile in the proportion of 60:40 v/v, flow rate of 1.0 ml/min and a detection wavelength of 240 nm using a UV detector.
Results: The developed method resulted in Mebeverine eluting at 5.8 min and Chlordiazepoxide at 3.9 min. Mebeverine exhibited linearity in the range 250-750Î¼g/ml, while Chlordiazepoxide exhibited linearity in the range 9.25-27.75Î¼g/ml. The precision is exemplified by relative standard deviations of 1.45% for Mebeverine and 1.58% for Chlordiazepoxide. Percentage Mean recoveries were found to be in the range of 98â€102, during accuracy studies. The limit of detection (LOD) for Mebeverine hydrochloride and Chlordiazepoxide were found to be 91.67Âµg/ml and 2.23Âµg/ml respectively, while limit of quantitation (LOQ) for Mebeverine hydrochloride and Chlordiazepoxide were found to be 277.81Âµg/ml and 6.77Âµg/ml respectively.
Conclusion: A simple, accurate, precise, linear and rapid RP-HPLC method was developed for simultaneous quantitative estimation of Mebeverine hydrochloride and Chlordiazepoxide in MEVA C capsules and validated as per ICH guidelines. Hence it can be used for the routine analysis of Mebeverine and Chlordiazepoxide in capusles in various pharmaceutical industries.
2. Walash MI, Sharaf El-din MMK, El-enany NM, Eid MI, Shalan SM. Simultaneous determination of sulpiride and mebeverine by HPLC method using fluorescence detection: application to real human plasma. Chem Cent J 2012;6(13):1-12.
3. Souri E, Aghdami AN, Adib N. A stability indicating HPLC method for determination of mebeverine in the presence of its degradation products and kinetic study of its degradation in oxidative condition. Res Pharm Sci 2014;9(3):199-206.
4. Gupta SP, Upmanyu N, Garg G. Development and validation of spectrophotometric, HPTLC and HPLC methods for the determination of Imipramine and Chlordiazepoxide in pharmaceutical dosage forms. Der Pharm Sin 2012;3(2):185-92.
5. Sujatha N, Pavani KH. Analytical method development and validation of amitriptyline hydrochloride and chlordiazepoxide in tablet by RP-HPLC. Indian J Res Pharm Biotech 2013;1(5):655-9.
6. De Schutter JA, De Croo F, Vander Weken G, Venden Bossche W, De Moerloose P. Stability tudy and quantitative determination of mebeverine hydrochloride in tablets by means of eversed-phase high-performance liquid chromatography. Chromatographia 1985;20:185-92.
7. Al-Deeb Q, Al-Hadiya BM, Foda NH. Quantitative analysis of mebeverine in dosage forms by HPLC. Chromatographia 1997;44:427-30.
8. Dubois JG, Atassi G, Hanocq M. High-performance liquid chromatographic determination of chlordiazepoxide, its metabolises and oxaziridines generated after UV irradiation. J Chromatogr A 1994;662(2):255-62.
9. Roberts SE, Delaney MF. Determination of chlordiazepoxide, its hydrochloride and related impurities in pharmaceutical formulations by reversed-phase high-performance liquid chromatography. J Chromatogr 1984;283:265-72.
10. Greizerstein HB, Mclaughlin IG. The high-pressure liquid chromatographic determination of chlordiazepoxide and its n-demethyl metabolite in mouse brain. J Liq Chromatogr 1980;3(7):1023-30.
11. Puglisi CV, Desilva JAF. Determination of chlordiazepoxide and its metabolites in plasma by high pressure liquid chromatography. Anal Lett 1978;11(2):135-60.
12. Kohlhof K. Determination of chlordiazepoxide in mouse plasma by gas chromatographyâ€”negative-ion chemical ionization mass spectrometry. J Chromatogr B Biomed Appl 1994;660(1):95-101.
13. Sun SR. Quantitative determination of chlordiazepoxide and its metabolites in serum by fluorescence TLC-densitometry. J Pharm Sci 1978;67(5):639-41.
14. El Walily AFM, El Gindy A, Bedair MF. Application of first-derivating UV spectrophotometry, TLC-densitometry and liquid chromatography for the simultaneous determination of mebeverine hydrochloride and sulpiride. J Pharm Biomed Anal 1999;21:535-48.
15. Shama SA, Amin AS. Spectrophotomertric microdetermination of nefopam, mebeverine and phenylpropanolamine hydrochloride in pharmaceutical formulations using alizarins. Spectrochim Acta A Mol Biomol Spectrosc 2004;60:1969-74.
16. Zayed SI. Simultaneous determination of mebeverine hydrochloride and sulpiride using the first derivatives of ratio spectra and chemometric methods. Anal Sci 2005;21:985-9.
17. El-Didamony AM. Spectrophotometric determination of benzydamine HCl, levamisole HCl andmebeverine HCl through ion-pair complex formation with methyl orange. Spectrochim Acta A Mol Biomol Spectrosc 2008;69:270-5.
18. Abdelaleem EA, Abdel wahah NS. Validated chromatographic and spectrophotometric methods for analysis of some amoebicide drugs in their combined pharmaceutical preparation. Pak J Pharm Sci 2013:26(1):175-83.
19. Naouib RA, Abdelkauy M. Development and validation of stability indicating HPLC and HPTLC method for determination of sulpiride and mebeverine hydrochloride in combination. Eur J Med Chem 2010;45(9):3719-25.
20. Patel S, Patel NJ. Spectrophotometric and chromatographic simultaneous estimation of amitriptyline hydrochloride and chlordiazepoxide in tablet dosage form. Indian J Pharm Sci 2009;71(4):472-6.
21. Patel SK, Patel NJ. Simultaneous RP-HPLC estimation of trifluoperazine hydrochloride and chlordiazepoxide in tablet dosage forms. Indian J Pharm Sci 2009;71(5):545-7.
22. Patel S, Patel NJ, Patel SA. Simultaneous Spectrophotometric Estimation of Imipramine ydrochloride and Chlordiazepoxide in Tablets. Indian J Pharm Sci 2009;71(4):468-72.
23. Venisetty RK, Kamarapu SK. RP-HPLC Method development and validation for simultaneous estimation of clidinium bromide, chlordiazapoxide and dicyclomine hydrochloride in bulk and combined tablet dosage forms. Int J Ad Biomed Pharm Res 2013;2(1):35-40.
24. Haggag RS, Shaalan RA, Belal TS. Validated HPLC determination of two fixed dosage combinations (chlordiazepoxide hydrochloride and mebeverine hydrochloride, carvedilol and hydrochlorothiazide) in their tablet. J AOAC Int 2010;93(4):1192-200.
25. International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human use. Validation of Analytical Procedures: Text and Methodology ICH Q2 (R1); 2005.