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Abstract

Paper presents a novel high performance liquid chromatography method development, optimization, validation and application of determining blonanserin in bulk drug, its formulation, human serum and in the presence of its degradation products. Selection of important factors which may influence separation were concluded: percentage of acetonitrile in the mobile phase, the percentage of orthophosphoric acid in aqueous phase and flow rate. Factors selected were altered in accordance to Box-Behnken design plan and investigation in factors having significant influence on separation were executed. Finally, the choicest values from solution offered of investigated factors were 43 % of acetonitrile in the mobile phase, 0.03% orthophosphoric acid in water and flowrate of 0.75 ml/min. The method validation was performed according to International Conference on Harmonisation guideline acceptance criteria for robustness, linearity, selectivity, precision and accuracy. Quantitative and qualitative system responses have not significantly affected by little variations in chromatographic parameters, which demonstrates robustness of the method. Limits of quantification and detection for blonanserin was calculated. Method shows excellent linearity, which was confirmed by r 2 = 0.9991 over the concentration range of 5 -50 µg/ml. Accuracy was calculated by recovery experiment and found to be in range 99.44-100.76 %. Precision was carried out at different levels: analysis repeatability, injection repeatability and intermediate precision and was found well under acceptable criteria. Furthermore, the applicability of method was analysed in real time sample analysis of blonanserin in its commercial pharmaceutical preparations and in-vitro human serum samples.