RP-HPLC ASSAY METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS ESTIMATION OF DILOXANIDE FUROATE AND ORNIDAZOLE IN TABLETS
Objective: To develop an accurate, precise and linear Reverse Phase High Performance Liquid Chromatographic (RP-HPLC) method for simultaneous quantitative estimation of Diloxanide furoate and Ornidazole in tablets and validate as per ICH guidelines.
Methods: The optimized method uses a reverse phase column, Waters Symmetry C18 (250 X 4.6 mm; 5Î¼), a mobile phase of potassium dihydrogen ortho phosphate buffer (20 mM, pH2.3): acetonitrile in the proportion of 40:60 v/v, flow rate of 1.0 ml/min and a detection wavelength of 230 nm using a UV detector.
Results: The developed method resulted in Diloxanide furoate eluting at 5.85 min and Ornidazole at 2.85 min. Diloxanide furoate exhibited linearity in the range 22.5-67.5Î¼g/ml, while Ornidazole exhibited linearity in the range 15-45Î¼g/ml. The precision is exemplified by relative standard deviations of 1.58% for Diloxanide furoate and 1.53% for Ornidazole. Percentage Mean recoveries were found to be in the range of 95â€105 by percentage method during accuracy studies.
Conclusion: A rapid, simple, accurate, precise and linear RP-HPLC method was developed for simultaneous quantitative estimation of Diloxanide furoate and Ornidazole in tablets and validated as per ICH guidelines. Hence it can be used for the routine analysis of Diloxanide furoate and Ornidazole in tablets in various pharmaceutical industries.
2. Khattab FI, Ramadan NK, Hegazy MA, Ghoniem NS. Stability indicating methods for the determination of Ornidazole in the presence of its degradate according to ICH Guidelines. Pharm Anal Acta 2012;3:179:1-8.
3. Ogrendik M. Treatment of rheumatoid arthritis with ornidazole: a randomized, double-blind, placebo-controlled study. Rheumatol Int 2006;26:1132-7.
4. Faubion WA, Bousvaros JA. Medical therapy for refractory pediatric Crohnâ€™s disease. Clin Gastroenterol Hepatol 2006;4:1199-213.
5. Danao KR, Hiradeve SM, Moon RS, Kasture AV, Yeole PG. RP-HPLC simultaneous estimation of Metronidazole and Diloxanide furoate in combination. Int J Pharm Life Sci 2010;1:82-5.
6. McAuley JB, Herwaldt BL, Stokes SL, Becher JA, Roberts JM, Michelson MK, et al. Diloxanide furoate for treating asymptomatic entamoeba histolytica Cyst Passers: 14 Years Experience in the United States. Clin Infect Dis 1992;15:464-8.
7. Marsden PD. Treatment of cyst passers. Clin Infect Dis 1992;15:559.
8. Ata AH, El-Haieg MO, Aboul-Magd LA, Mansour AH. A comparative study on some nti-amoebic drugs in cyst-passers. J Egypt Soc Parasitol 1984;14:213-8.
9. Soma SM, Vidyasagar V, Narsaiah N, Anandkumar R, Krishna DR. Validated HPLC method for the determination of Ornidazole in human serum and urine. Indian J Pharm Sci 2005;67:302-6.
10. Groppi A, Papa P, Montagna M, Carosi G. Determination of Ornidazole in human plasma and RBCs using HPLC. J Chromatogr 1986;380:437-42.
11. Nagavallai D, Sankar ASK, Ananda kumar K, Karunambigai K, Raju MSSN. RP-HPLC method for simultaneous estimation of Gatifloxacin and Ornidazole in tablets. Indian J Pharm Sci 2007;69:333-5.
12. Gandhimathi M, Ravi TK, Nilima S. A validated HPLC method for simultaneous estimation of Ofloxacin and Ornidazole in tablet dosage form. Indian J Pharm Sci 2006;68:838-40.
13. Dhandapani B, Thirumoorthy N, Shaik Harun R, Rama kotaiah M, Anjaneyalu N. Method development and validation for the simultaneous estimation of Ofloxacin and Ornidazole in tablet dosage form by RP-HPLC. Int J Pharm Sci Res 2010;1:78-83.
14. Paramane S, Kothapalli L, Thomas A, Despandey AD. Simultaneous RP-HPLC estimation of Gatifloxacin and Ornidazole in tablet dosage forms. Indian J Pharm Sci 2007;69:525-8.
15. Ranjane PN, Gandhi SV, Kadukar SS, Ranher SS. Simultaneous determination of Cefuroxime axeitil and Ornidazole in tablet dosage form by using RP-HPLC. Chin J Chromatogr 2008;26:763-5.
16. Natarajan S, Raman B. Development and validation of a stability indicating HPLC method for simultaneous estimation of Ofloxacin and Ornidazole. Indian Pharm 2005;4:79-84.
17. Pai P, Rao GK, Srinivas B, Puranik S. RPLC determination of Tinidazole and Diloxanide furoate in tablets. Indian J Pharm Sci 2008;70:670-2.
18. Satyabrata R. HPLC determination of Metronidazole and Diloxanide furoate in single and combined formulation. East Pharm 1994;2:131-3.
19. Rao GR, Murthy SSN, Mohan K. HPLC assay of Diloxanide furoate and Metronidazole in combined dosage forms. Indian Drugs 1983;21:455-7.
20. Gizawy EISM. HPLC analysis of Metronidazole and Diloxanide furoate in its dosage forms. Anal Lett 1995;28,83-92.
21. Mishal A, Sober D. Stability indicating reversed-phase liquid chromatographic determination of metronidazole benzoate and diloxanide furoate as bulk drug and in suspension dosage form. J Pharm Biomed Anal 2005;39:819-23.
22. Shaalan Al, Nora H. Determination of diloxanide furoate and metronidazole in binary mixture using first derivative of the ratio spectra and high-performance liquid chromatography-UV methods. Am J Appl Sci 2007;4:66-72.
23. Sivaprasad P, Yaswanth kumar S, Ashok Kumar A. RP-HPLC method development and validation for simultaneous quantitative estimation of Diloxanide furoate and Ornidazole in tablets. Int J Pharm Pharm Sci 2015;7:515-9.
24. Vijay kumar D, Swetha P, Ashok kumar A. Assay method development and validation for simultaneous quantitative estimation of Diloxanide furoate and Ornidazole in tablets by RP-HPLC. Int J Pharm Pharm Sci 2015;7. [Article in Press]
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.