Development and Validation of Bioanalytical Method for Simultaneous Quantification of Anti-Hypertensive Drugs by Using HPLC

Abstract

The chromatographic separation was achieved by using eclipse plus C8 (250mm x 4.6mm, 5μm) column by applying isocratic elution using the mobile phase Ⅰ consist of (methanol + acetonitrile 60:40 v/v) and mobile phase Ⅱ consist of (water with 0.01% of diethyl amine) in the ratio 80: 20 (v/v) detection was carried at the 225 nm by employing 1ml/min flow rate. The separation of drugs from the plasma was achieved using protein precipitation technique by acetonitrile. The retention time was found to be 6.573 min and 5.421 min for metoprolol succinate and cilnidipine respectively. The developed method has the linearity range was obtained for RP-UFLC method was 0.5 -16 µg/ml and 1-32 µg/ml for cilnidipine and metoprolol succinate respectively and the regression co-efficient (r 2) is 0.999 for the both drugs. Accuracy & precision is evaluated for the method and found be within the limit and the results were reproducible.