Validated HPLC method for the determination of nocodazole in solid lipid nanoparticle formulations [Kati{dotless} lipid nanopartikül formiilasyonlan İindeki nokodazol'ün belirlenmesiiçin onaylanmiş HPLC metodu]

Abstract

Nocodazole (NCD) is an anti-neoplastic agent which exerts its effect in cells by interfering with the polymerization of microtubules. As NCD affects the cytoskeleton, it is often used incell biology experiments as a control. In this study, NCD was loaded into Solid Lipid Nanoparticle (SLN) systems. A reverse phase high performance liquid chromatography (HPLC) method was validated and applied forthe determination of NCD in SLN. Determination and validation studies were carried out on a 4.6 × 150 mm, 5µm C18 Thermo column using an optimized mobile phase (MP) of methanol:water:phosphate buffer (45:42.5:12.5, v/v/v, pH 5.03) at a flow rate of 0.8 mL/min. Diode array detection was performed at 256 nm and the column temperature was adjusted to 40°C. Naproxen was used as an internal standard (IS). The retention times for naproxen and NCD were 2.2 and 9.8 min, respectively. The specified working range was derived from linearity studies and kept in the concentration range 0.5-100 ppm. Limit of dedection (LOD) and limit of quantitation (LOQ) values were determined to be 0.065 ppm and 0.196 ppm, respectively. NCD recovery % results of the SLN formulations which are stored at 25°C, 4°C and 40°C were investigated and compared to the freshly prepared samples.