Original Research Article
Year: 2019 | Month: July-September | Volume: 4 | Issue: 3 | Pages: 160-165
Development and Validation of Novel RP-HPLC Method for the Simultaneous Estimation of Amlodipine and Hydrochlorothiazide in Combined Dosage Form
Nagaraju Pappula, Koppula Jyothi, Shaik Ayesha Ameen
Department of Pharmaceutical Analysis, Hindu College of Pharmacy, Amaravathi Road, Guntur – 522 002, A.P.
Corresponding Author: Nagaraju Pappula
ABSTRACT
A simple, specific, accurate, rapid, inexpensive Reversed Phase High Performance liquid chromatographic method has been developed and validated for the simultaneous determination of Amlodipine and Hydrochlorothiazide in pharmaceutical tablet dosage form. Separation was achieved using a C18 column with mobile phase consisting of Phosphate Buffer (pH 3.4) and Methanol (55:45 v/v) in isocratic mode at 1 ml/min flow rate. Column effluent was monitored at 238 nm using a UV detector. The retention time for Amlodipine and Hydrochlorothiazide was found to be 2.7 and 3.4 min respectively the method was validated for System Suitability, Specificity, linearity, accuracy, precision, robustness. The method demonstrated excellent linearity for Amlodipine and Hydrochlorothiazide with regression coefficients of 0.9999 and 0.9998, respectively. Linearity was established for Amlodipine besylate in the range of 20-80 µg / mL and Hydrochlorothiazide in the range of 50-200 µg / ml. The accuracy of the method is evaluated in triplicate at three concentration levels i.e. 50%, 100% and 150% of target test concentration. The method was found to be sensitive with quantification limits of 1.05 and 2.82 µg/ml for Amlodipine and Hydrochlorothiazide. The method was successfully employed for the determination of Amlodipine and Hydrochlorothiazide in commercially available tablets.
Keywords: Amlodipine, Hydrochlorothiazide, RP-HPLC, UV-Vis Detector, Validation