SIMULTANEOUS ESTIMATION OF BEPOTASTINE BESILATE AND BENZALKONIUM CHLORIDE IN OPHTHALMIC FORMULATION BY RP-HPLC METHOD
Objective: Development and validation of stability indicating RP-HPLC method for the simultaneous determination of bepotastine besilate (Bepo B) and benzalkonium chloride (BKC) in an ophthalmic dosage form.
Methods: A chromatographic separation of the drug, as well as a preservative, was achieved using Shimadzu HPLC 1100 series consisted of binary pump LC-10 ADvp, Rheodyne universal injector 7725i and Shimadzu SPD-10 UVâ€“Visible detector. The chromatographic separations were performed using AnalyticalÂ® Hyperchrome ODS C18, 5 Î¼m, 250 mm X 4.6 mm i.d. column with isocratic mobile phase Acetonitrile: phosphate buffer (60:40) pH 5.5. The drug and a preservative were monitored at an ambient temperature and detection wavelength of 210 nm with a flow rate of 1 ml/min and an injection volume of 20 Âµl.
Results: The mean % recovery at the 80, 100 and 120% level for Bepotastine and benzalkonium chloride was found to be 100.09 and 100.81% respectively and % RSD was found to be 0.21 and 0.85% respectively, which meets the established acceptance criteria. Forced degradation of bepotastine besilate was carried under alkaline, acidic, neutral, oxidative, humidity, thermal and photodegradation conditions and it was analyzed by proposed method. The drug degrades to some extent in all forced degradation condition.
Conclusion: The developed method was validated as per ICH guidelines using validation parameters such as accuracy, precision, linearity and range, robustness, ruggedness, LOD, LOQ, specificity, and system suitability testing. The proposed method can be used for routine analysis stability testing and assay of bepotastine besilate ophthalmic solution in quality control laboratories.
Keywords: Bepotastine besilate (Bepo B), Benzalkonium chloride (BKC)
2. Yato N, Murata T, Saito N, Sakai A, Kikuchi M, Tsuzurahara K, et al. the Anti-allergic activity of Bepotastine Besilate (TAU-284), a new anti-allergic drug. Folia Pharmacol Jpn 1997;110:19-29.
3. Craig FM, Shannon EM. Comparative efficacy of bepotastine besilate 1.5% ophthalmic solution versus olopatadine hydrochloride 0.2% ophthalmic solution evaluated by patient preference. Clin Ophthalmol 2012;6:1731-8.
4. http://www.accessdata.fda.gov/drugsatfda_docs/nda/2009/022288s000_SumR.pdf. [Last accessed on 10 Jan 2014].
5. Yang YW, Zhang WQ, Zhu Y. Simultaneous determination of quaternary ammonium salts in antiseptics by high-performance liquid chromatography. Chin J Health Lab Technol 2006;16:823-24.
6. Prince SJ, McLaury HJ, Allen LV, McLaury P. Analysis of Benzalkonium chloride and Its homologs: HPLC versus HPCE. J Pharm Biomed Anal 1999;19:877-82.
7. Mamta PD, Anandan P, Arindam M. A rapid and sensitive method for estimation of Bepotastine in human plasma by LCMS/MS. Int J Pharm Biol Sci 2011;2:291-7.
8. Sharath KP, Prashant LP. Method development and validation of bepotastine besilate by RP-HPLC. Inventi Rapid: Pharm Anal Quality Assurance 2013;3:1-4.
9. Narasimha RK, Nagaraju VS, Rajan ST, Eshwaraiah S, Sampath RG, Rakesh M. Stability indicating HPLC method for the quantification of bepotastine besilate and its related substances. Der Pharm Chem 2014;6:343-51.
10. ICH Q1A (R2). Stability testing of new drug substances and products; 2003.