DEVELOPMENT AND VALIDATION OF AN RP-HPLC METHOD FOR QUANTITATIVE ESTIMATION OF ILOPERIDONE IN RAT PLASMA

A simple and fast isocratic reverse phase- high performance liquid chromatography (RP-HPLC) method was developed for estimation of iloperidone from rat plasma employing a photo diode array (PDA) detector. The plasma samples were prepared by protein precipitation using acetonitrile as extracting solvent. The chromatographic separation was performed on a BDS Hypersil Phenyl column (250 mm × 4.6 mm; 5 μm) using a mixture of 0.01 M sodium dihydrogen phosphate buffer (pH 6.0) and acetonitrile (60:40, v/v) as mobile phase with a flow rate of 1.5 ml/min, retention time 7.04 min, and detection wavelength 276 nm. The method was validated for linearity, sensitivity, selectivity, accuracy, precision, and stability according to USFDA guidelines. The calibration curve exhibited linearity in the concentration range of 50–1000 ng/ml (r²=0.998) and the lower limit of quantification (LLOQ) and limit of detection (LOD) of the method were 50 ng/ml and 30 ng/ml respectively. The chromatographic runs were selective without any interfering peaks at the retention time of iloperidone. The accuracy and precision of the method were within USFDA specified range. The plasma samples showed stability over a period of 24 h at room temperature, for two months at – 20 °C, and three freeze/thaw cycles.