RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS QUANTITATIVE ESTIMATION OF DILOXANIDE FUROATE AND TINIDAZOLE IN TABLETS
Keywords:
RP-HPLC, Diloxanide furoate, Tinidazole, Method development, ValidationAbstract
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 Tinidazole in tablets and validate as per ICH guidelines.
Methods: The optimized method uses a reverse phase C18 column, ZODIAC (250 X 4.6 mm; 5μ), mobile phase consisting of mixed phosphate buffer (pH 6.5; KH2PO4+K2HPO4): acetonitrile in the proportion of 30:70 v/v. The mobile phase was set at a flow rate of 1.0 ml/min and the volume injected was 20μl for every injection. The detection wavelength was set at 270 nm.
Results: The developed method resulted in Diloxanide furoate eluting at 4.70 min and Tinidazole at 3.45 min. Diloxanide furoate exhibited linearity in the range 30-70μg/ml, while Tinidazole exhibited linearity in the range 36-84μg/ml. The precision is exemplified by relative standard deviations of 0.266% for Diloxanide furoate and 0.35% for Tinidazole. Percentage Mean recoveries were found to be in the range of 98â€102, during accuracy studies. The limit of detection (LOD) for Diloxanide furoate and Tinidazole were found to be 0.32µg/ml and 0.40µg/ml respectively, while limit of quantitiation (LOQ) for Diloxanide furoate and Tinidazole were found to be 0.96µg/ml and 1.21µg/ml respectively.
Conclusion: A simple, accurate, precise, linear and rapid RP-HPLC method was developed for simultaneous quantitative estimation of Diloxanide furoate and Tinidazole in tablets and validated as per ICH guidelines. Hence it can be used for the routine analysis of Diloxanide furoate and Tinidazole in tablets in various pharmaceutical industries.
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Sirisha T, Gurupadayya BM, Sridhar S. Simultaneous determination of Ciprofloxacin and tinidazole in tablet dosage form by reverse phase high performance liquid chromatography. Trop J Pharm Res 2014;13(6):981-7.
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(2):82-5.
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(3):464-8.
Marsden PD. Treatment of cyst passers. Clin Infect Dis 1992;15(3):559.
Ata AH, El-Haieg MO, Aboul-Magd LA, Mansour AH. A comparative study on some anti-amoebic drugs in cyst-passers. J Egypt Soc Parasitol 1984;14(1):213-8.
Walash MI, Ibrahim F, El Abass S. Isocratic RP-HPLC method for separation and simultaneous determination of ternary mixture of Omeprazole, Tinidazole and Doxycycline in their raw materials and combined capsules. Anal Methods 2013;5:5105-11.
Sebaiy MM, El-Shanawany AA, Adl SM, Abdel-Aziz LM, Hashem HA. Rapid RP-HPLC method for simultaneous estimation of Norfloxacin and Tinidazole in tablet dosage form. Asian J Pharm Ana 2011;1(4):79-84.
Vikas K, Senthil kumar M, Mahadevan N. Simultaneous estimation of Amoxicillin, Tinidazole and Omeprazole in microsphere formulation by RP-HPLC. Int J Rec Adv Pharm Res 2012;2(2):78-83.
Meshram DB, Bagade SB, Tajne MR. Simple HPLC method for simultaneous estimation of Fluconazole and Tinidazole in combined dose tablet. J Chromatogr Sci 2009;47(10):885-8.
Chiranjeevi B, Swati B, Kalyan O. Novel reverse phase HPLC method development and validation of Fluconazole and Tinidazole in a combined tablet dosage form. Int J Chem Tech Res 2011;3(3):1309-17.
Mahmoud MS, Abdullah AS, Sobhy MEA, Lobna MAA, Hisham AH. Rapid RP-HPLC method for simultaneous estimation of Norfloxacin and Tinidazole in tablet dosage form. Asian J Pharm Anal 2011;1(4):79-84.
Hackett LP, Dusci LJ. Determination of Metronidazole and Tinidazole in human plasma using high performance liquid chromatography. J Chromatogr 1979;175(2):347-9.
Satyabrata R. HPLC determination of Metronidazole and Diloxanide furoate in single and combined formulation. East Pharm 1994;2:131-3.
Rao GR, Murthy SSN, Mohan K. HPLC assay of Diloxanide furoate and Metronidazole in combined dosage forms. Indian Drugs 1983;21:455-7.
Gizawy EISM. HPLC analysis of Metronidazole and Diloxanide furoate in its dosage forms. Anal Lett 1995;28:83-92.
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(3-4):819-23.
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 App Sci 2007;4(2):66-72.
Pai P, Rao GK, Srinivas B, Puranik S. RPLC determination of Tinidazole and Diloxanide uroate in tablets. Indian J Pharm Sci 2008;70:670-2.
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.