Development of a HILIC method for the determination of 5-fluorouracil from nano drug delivery systems and rat skin extracts

•First hydrophilic interaction liquid chromatography with diode array detection study for stability indicating determination of 5-Fluorouracil.•5-Fluorouracil loaded solid lipid nanoparticle and nano lipid carrier formulations were developed.•Developed method has sufficient sensitivity for the estimation of 5-Fluorouracil in raw materials, marketed formulation and rat skin extracts.•Ex-vivo Penetration/Permeation Studies with rat skin indicated that higher dermal accumulation of 5-Fluorouracil was obtained with nano lipid carrier formulation. This is the first report in literature using hydrophilic interaction liquid chromatography (HILIC) in combination with diode array detector (DAD) for stability indicating determination of 5-Fluorouracil (5-FU) from its bulk form, pharmaceutical preparations, developed solid lipid nanoparticle (SLN) and nano structured lipid carrier (NLC) drug delivery systems as well as the rat skin extracts. The separation was performed at 45 °C, on Sequant Zic HILIC (250 mm × 4.60 mm ID, 5 μm, 200 Ao), peek HPLC column. Mobile phase is consisting of a mixture of acetonitrile: buffer containing 5 mM ammonium acetate (95:5; v/v). The pH of the mobile phase was adjusted to 7.0 using 1 M NaOH. The analysis was carried out at 0.75 mL min−1 flow rate with a detection wavelength of 265 nm and the injection volume was arranged as 10 μL. The developed method was fully validated in accordance with the International Council on Harmonization (ICH) Guidelines. Specificity of this method was demonstrated by forced degradation studies. As a result of calibration studies, the calibration curve was found linear in the concentration range of 1–250 μg mL−1 (R2 = 0.999). The precision of this technique calculated within the frame of intra-day and inter-day based on a percentage of relative standard deviation (RSD%) values (