VALIDATED STABILITY-INDICATING RP-HPLC ASSAY METHOD FOR AZATHIOPRINE IN PHARMACEUTICAL DOSAGE FORM ACCORDING TO ICH GUIDELINES
Keywords:Azathioprine, Assay HPLC, Validation, Stability-indicating, Degradation profile
Objective: A simple, sensitive, precise and accurate stability-indicating HPLC method has been developed and validated for determination of Azathioprine in bulk drug and in pharmaceutical dosage form in the presence of degradation products.
Methods: An isocratic, reversed phase HPLC method was developed to separate the drug from the degradation products, using a Hypersil C8, 150 X 4.6 mm, 5Âµ, Thermostat column compartment connected with Waters (alliance) Empower software. Mobile phase consists mixture of Potassium dihydrogen phosphate(A) and Acetonitrile(B) in the ratio (pH 6.50Â±0.1, ratio 60:40 v/v) at a flow rate of 1.0 mL /min. The linear regression analysis data for the calibration curve showed a good linear relationship with regression coefficient 1. The detection was carried out at a wavelength of 254 nm.
Results: The linearity of the method were excellent over the range 2.5-15 Î¼g/ml, the linear regression equation was Y=76621Ã—+12515. The Azathioprine was subjected to stress conditions of hydrolysis (acid, base), photolysis and thermal degradation. Degradation was observed for Azathioprine in acid, base, heat and UV. The degradation products were well resolved from the main peak. The percentage recovery of Azathioprine was from (98.0 to 102.0 %.) in the pharmaceutical dosage form.
Conclusion: The results demonstrated that the method would have a great value when applied in quality control and stability studies. The developed method was validated with respect to linearity, accuracy (recovery), precision, system suitability, specificity and robustness. The forced degradation studies prove the stability indicating power of the method.
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