European Journal of Biomedical and Pharmaceutical Sciences

A VALIDATED REVERSE PHASE-HPLC METHOD FOR QUANTIFICATION OF RISPERIDONE IN RAT PLASMA BY PDA DETECTOR

Vineet Dubey* and Tulsi Ram Saini

ABSTRACT

A reverse phase-high performance liquid chromatographic method was developed using photo-diode array detector for the quantification of risperidone in rat plasma. The Hypersil BDS Phenyl column (4.6 mm × 250 mm, 5 μm) was used for separation with a mixture of acetonitrile and 0.01 M sodium dihydrogen phosphate buffer (pH 6) as mobile phase in 60:40 ratio at a flow rate of 1 ml/min. The chromatographic analysis was performed in isocratic mode at 280 nm detection wavelength with a total run time of 7 min and a retention time of 6.1-6.3 min. The protein precipitation method was employed to prepare rat plasma samples for chromatographic analysis. Linearity, sensitivity, accuracy, precision, selectivity, and stability of the developed method were determined in accordance with the USFDA bioanalytical method validation guidelines. The method showed linearity over 100–5000 ng/ml concentration range with 60 ng/ml as limit of detection and 100 ng/ml as limit of quantitation. The accuracy was within a relative error of 6% of the nominal drug concentrations and precision was under 7% coefficient of variance for intra-day as well as inter-day analyses. Stability of plasma samples was also acceptable as per USFDA guidelines. The selectivity of the method for risperidone was confirmed by the absence of any interfering peaks at the retention time of the drug.

Keywords: Reverse phase-HPLC method, PDA detector, risperidone, rat plasma, validation.