STABILITY INDICATING RP-HPLC AND HPTLC METHODS FOR THE DETERMINATION OF ZANAMIVIR IN BULK AND DOSAGE FORM
Keywords:
Zanamivir, Reverse phase high performance liquid chromatography, High pressure thin layer chromatography, Virenza capsule, Method development and validation, Stability indicatingAbstract
Objective: To develop a simple, precise, accurate, validated stability indicating reverse phase high-pressure liquid chromatography and high-performance thin layer chromatography method for the determination of zanamivir in bulk and capsule dosage form.
Methods: The high-pressure liquid chromatography separation was achieved on Agilent TC C18 (2) 250 x 4.6 mm, 5 μ column using mobile phase composition of methanol-0.02 M phosphate buffer, pH 3.5, 50:50 (v/v). Flow rate was maintained at 1 ml/min at an ambient temperature. Quantification was achieved with ultraviolet detection at 230 nm. The retention time obtained for zanamivir was at 3.6 min. The method employed thin layer chromatography aluminum plates precoated with silica gel 60F-254 as the stationary phase. The solvent system consisted of chloroform: methanol: acetic acid (4.5:0.5:0.3v/v) and then scanned. The system was found to give a compact spot for zanamivir (Rf value of 0.29±0.02).
Results: In HPLC,the result obtained with the detector response was found to be linear in the concentration range of 2-12 μg/ml. In HPTLC, the linear regression analysis data for the calibration plots showed a good relationship with r2=0.9999±0.0001 in the concentration range 500-3000 ng/spot. The reliability and analytical performance of the proposed methods, including linearity, range, precision, accuracy, detection and quantitation limits, were statistically validated. When zanamivir was subjected to different stress conditions; the proposed methods could effectively separate the drug from its degradation products, and were thus considered as good stability-indicating procedures.
Conclusion: It is concluded that this method can be applied for routine quality control of zanamivir in dosage forms as well as in bulk drug.
Keywords: Zanamivir, Reverse phase high-performance liquid chromatography, High-pressure thin layer chromatography, Virenza capsule, Method development and validation, Stability indicating
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