Validation and application of a modified RP-HPLC method for the quantification of midazolam in pharmaceutical dosage forms.

ABSTRACT

The purpose of the study was to develop a simple, sensitive and rapid RP-HPLC method for the determination of Midazolam in marketed products. Chromatographic determination was performed in a reverse phase C18 column (250 mm × 3.3 mm I.D. , 5μm particle size) using a mixture of acetonitrile methanol 0.065 M ammonium acetate buffer (502030, v/v/v), final pH adjust to 5.5 ± 0.02 with ortho phosphoric acid as mobile phase and delivered at a flow rate of 1 ml/min. The UV detection was set at 220 nm. The calibration range was from 2.0 to 30.0 μg/ml. The method was validated in term of linearity (r2>0.98, RSD= 1.958%), precision (RSD=3.757 %) and accuracy. The limit of quantification was 2 μg/ml and the limit of detection was 0.1 μg/ml. The potency of midazolam in marketed products was determined by this method with acceptable precision and reproducibility.