A VALIDATED RP-HPLC ASSAY METHOD FOR DETERMINATION OF GEMCITABINE LOADED NANOSIZED SOLID LIPID NANOPARTICLES

Abstract

A novel reverse phase HPLC assay method has been developed and validated for the simultaneous determination of Gemcitabine Hydrochloride (dFdCH) along with solid lipid nanoparticles SLNs and conjugated with mannose to targeting the lungs for chemotherapy. Methanol was used as the extracting solvent for preparation of tissue sample. Methanol: Ammonium acetate buffer; 10:90 v/v (pH 5) was the mobile phase at flow rate 1.5 mL/min at pressure of 102/101 bars using Luna Phenomenex, C18 (4.6mmA—250 mm; 5 µm bead size) at wavelength 269 nm. The column oven temperature was optimized at 350C. The biodistribution studies were conducted to evaluate the target potential at the sites of interest in liver, spleen, lung and kidney respectively, the calibration curve was found to be linear over the concentration range of 100-5000 ng/mL(r2=0.9980, r2=0.9980, r2=0.9990, r2=1 respectively). Plain drug have greatest access to liver and secondarily to spleen and then kidney but on the contrary the concentration of drug was greatest in lung when treated with mannosylated SLNs, data suggested that the mannose attachment has provided the access for the drug in the lungs via the formulation in greater quantity than free drug due to interaction of mannose with mannose receptors present on lung macrophages. The developed method was validated in accordance to ICH guidelines. Keywords: High Performance Liquid Chromatography, Mannosylated SLNs, Macrophage Mannose, Receptor (MMR), Lung Cancer, Gemcitabine Hydrochloride.