STABILITY-INDICATING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR DETERMINATION OF ANTIHISTAMINE DRUG AZELASTINE
Objective: The objective of this research was to develop a simple, precise, accurate, and stability-indicating reverse-phase high-performance liquid chromatographic method for estimation of azelastine hydrochloride (AZL) in nasal spray preparation.
Methods: Chromatography was performed on a 250 mmÃ—4.6 mm, 5-Î¼m particle size, Waters Spherisorb CN column using (50:50 v/v) mixture of potassium dihydrogen phosphate buffer and acetonitrile as mobile phase. The detection was carried out at 290 nm and flow rate employed was 1.0 ml/min. The degradation of AZL was studied under different ICH recommended stress conditions.
Results: The retention time was 4.34 min for AZL. Linearity was established in the concentration range of 5â€“120 Î¼g/ml, with a correlation coefficient of 0.9996. Limit of detection (LOD) and limit of quantitation (LOQ) were found to be 0.81 Î¼g/ml and 2.44 Î¼g/ml, respectively. Percentage recovery was found between 99 and 102%. The values of percentage relative standard deviation (<2%) proved the high precision of the proposed method. The method was found to be robust regarding any small variation in the column temperature, pH of mobile phase, and mobile phase ratio. AZL was found stable in 5 M HCl at 80Â°C for 5 h, 5 M NaOH at 80Â°C for 5 h, 30% H2O2 at 80Â°C for 5 h, and in oven at 70Â°C for 8 h.
Conclusion: The results obtained in this research work clearly proved that the proposed HPLC method for the assay of AZL in nasal spray preparation is simple, precise, specific, accurate, and stability indicating. It indicates that the method is suitable for analysis of AZL in the raw material and the pharmaceutical product without interference from excipients.
2. HMSO Publication. British Pharmacopoeia. Vol. 1. London: HMSO Publication; 2009. p. 488-91.
3. Krishnan PV. Current Index of Medical Specialities (CIMS). Vol. 116. Banglore: Bio Grad Medical Services; 2012.
4. Pivonka J, Segelman FH, Hartman CA, Segl WE, Kucharezyk N, Sofia RD. Determination of azelastine and desmethylazelastine in human plasma by high performance liquid chromatography. J Chromatography B Biomed Sci Appl 1987;420:89-98.
5. Park YS, Kim SH, Kim YJ, Yang SC, Lee MH, Shaw LM, et al. Determination of azelastine in human plasma by validated liquid chromatography coupled to tandem mass spectrometry (LC-ESI/MS/MS) for the clinical studies. Int J Biomed Sci 2010;6:120-7.
6. Langevin CN, Pivonka J, Wichmann JK, Kucharczyk N, Sofia RD. High performance liquid chromatographic determination of azelastine and desmethylazelastine in guinea pig plasma and lung tissue. Biomed Chromato 1993;7:7-11.
7. Zha W, Shum L. Simultaneous determination of azelastine and its major metabolite desmethylazelastine in human plasma using high performance liquid chromatography-tandem mass spectrometry. J Chromato B 2012;906:69-74.
8. Heinemann U, Blaschke G, Knebel N. Simultaneous enantio selective separation of azelastine and three of its metabolites for the investigation of the enantiomeric metabolism in rats: I. Liquid chromatographyâ€“ ionspray tandem mass spectrometry and electrokinetic capillary chromatography. J Chromato B 2003;793:389-404.
9. Salama NN, Abdel-Razeq SA, Atty SA, Kosy NE. Spectrophotometric determination and thermodynamic studies of the charge transfer complexes azelastine hydrochloride. Bull Fac Pharm Cario Univ 2011;49:13-8.
10. Wyszomirska E, Czerwinska K, Kublin E, Mazurek AP. Identification and determination of ketotifen hydrogen fumarate, azelastine hydrochloride, dimetindene maleate and promethazine hydrochloride by densitometric method. Acta Pol Pharmaceut 2013;70:951-9.
11. Boovizhikannan T, Palanirajan VK. RP-HPLC determination of azelastine in pure and in ophthalmic formulation. Int J Pharm Sci Rev Res 2012;17:62-4.
12. Patel RB, Patel NK. Development and validation of IR spectrophotometric method for estimation of azelastine hydrochloride in nasal spray preparations. Available from: http://www.pharmatutor. org/articles/development-validation-ir-spectrophotometric-method-estimation-azelastine-hydrochloride-nasalspray-preparations.
13. Rao KL, Reddy KP, Babu KS, Raju KS, Rao KV, Sharik JV. Simultaneous estimation of fluticasone propionate, azelastine hydrochloride, phenylethyl alcohol and benzalkonium chloride by rp-hplc method in nasal spray preparations. Int J Res Pharm Sci 2010;1:473-80.
14. Salama NN, Abdel-Razeq SA, Atty SA, Kosy NE. Development and validation of densitometry tlc stability indicating method for quantitative determination of azelastine hydrochloride and emedastine difumarate in their drug products. Br J Pharm Res 2014;4:79-92.
15. ICH Harmonised Tripartite Guideline Test on Validation of Analytical Procedures Q2A; 1994.
16. ICH Harmonised Tripartite Guideline Validation of Analytical Procedure: Methodology (Q2B); 1996.
17. Gul W. Photostability testing of formulated phaemaceutical products. Int J Pharm Pharm Sci 2015;2:65-8.
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.