Liquid Chromatographic Assay for the Analysis of Kanamycin sulphate nanoparticles in Rat after intramuscular administration: Application to a Pharmacokinetic Study

A rapid reversed-phase high performance liquid chromatography (RP-HPLC) method was developed for the determination of Kanamycin sulphate (KS) in PLGA nanoparticle formulation. A new formulation of KS loaded PLGA nanoparticles (NPs) was prepared by double (multiple) emulsion process in our laboratory. The desired chromatographic separation was achieved on a Phenomenex C18column under isocratic conditions using UV detection at 205 nm. The optimized mobile phase consisted of a mixture of 0.1 M disodium tetraborate (pH 9.0) and water (25:75, v/v) supplemented with 0.5 g/L sodium octanesulphonate at a flow rate of 1 mL/min. The linear regression analysis for the calibration curves showed a good linear correlation over the concentration range of 120-840µg/ml, with correlation coefficients of (r2 0.9997). The system was found to construct sharp peaks for KS and IS with retention times of 4.08 and 5.49 min, respectively. Transmission electron microscopy studies on MFX NPs demonstrated particle size < 100 nm. An average encapsulation efficiency of 74.34% was obtained for NPs. In vitro studies showed zero-order release and about 95% drug being released within 12 days in PBS (pH 7.4). In conclusion, the proposed optimized method was successfully applied for the determination of in vitro and in vivo release studies of KS NPs.