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Development and validation UPLC method for simultaneous determination of Hydrochlorothiazide and Eprosartan


Vanitha C., Bindu Pendota, Alagar Raja M., David Banji, Pradeep Kumar P

Abstract

Background: The objective of the study was to develop UPLC method for the quantitative determination of Eprosartan and Hydrochlorothiazide and its validation. UPLC is a better technique than HPLC in terms of performance and speed, so it was selected. The method was developed using Waters Aquity BEH shield RP C18 (2.1 x 100mm, 1.7μm) column with mobile phase containing a mixture of Acetonitrile and 0.05MPhosphate buffer (pH 4.5) in the ratio of 35:65 at a flow rate of 0.4ml/min. The eluted compounds were monitored at 245 nm, Results: The retention time of Hydrochlorothiazide and Eprosartan was found to be 0.902 min and 2.075 min respectively. The developed method was validated as per international conference on harmonization (ICH) guidelines with respect to linearity, limit of detection, limit of quantification, accuracy, precision and robustness. The precision was found to be within the limits. The linearity studies indicated the drug obeys Beer’s law and revealed the specified range of linearity for Hydrochlorothiazide was between 1.25-6.25μg/ml and for Eprosartan 25-125μg/ml. The robustness was observed from the insignificant variation in the analysis by changes in flow rate, mobile phase ratio. Conclusion: The rapid gradient UPLC method developed for quantitative analysis of Eprosartan and Hydrochlorothiazide in pharmaceutical dosage forms is precise, accurate, linear, robust and specific.




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