Development and validation of a reverse phase HPLC method for the estimation of lacidipine in rat plasma: Application to preclinical pharmacokinetics

A simple and reliable reverse phase high performance liquid chromatographic (RP-HPLC) method was developed for quantification of lacidipine in rat plasma using felodipine as internal standard. The protein precipitation technique was adopted for the sample preparation. Separation was carried out on a Hyperclone BDS C₁₈ column (250 × 4.6 mm, 5 μm). The mobile phase was comprised of acetonitrile and 25 mM ammonium acetate buffer (pH 4.5), (60:40 v/v), at the flow rate of 1.0 mL/min. The effluent was monitored using a UV detector at 240 nm. The retention time of lacidipine was found to be 19.97 min. The method was linear over the concentration range of 40-1800 ng/mL. The % RSD values of intra-day and inter-day precision were found to be less than 15 %. Recovery with respect to the area ratio was calculated at LQC, MQC and HQC levels. Stability studies showed that the lacidipine in plasma samples was stable. The method was successfully applied to preclinical pharmacokinetic study of lacidipine after oral administration of lacidipine loaded nanostructured lipid carriers in male Wistar rats.