Development and validation of an RP-HPLC/UV Method for Determination of Cholecalciferol in Polymeric Nanoparticles Suspensions / Desenvolvimento e validação de um método RP-HPLC/UV para determinação do Cholecalciferol em nanopartículas poliméricas Suspensões

Abstract

A simple and effective RP-HPLC/UV method was developed to determine cholecalciferol (COL) in suspensions of polymeric nanoparticles. Chromatographic conditions used in the methodology were a C18 RP column using an isocratic mobile phase of methanol and water (98:2 v/v) at a flow rate of 1.2 mL/min, UV detection at 265 nm. The validation parameters evaluated were: specificity, linearity, limit of quantification, limit of detection, precision, precision and robustness. The retention time was about 7.5 min. The method was linear, specific (r=0.9992) interval 10.0 to 80.0 μg/mL, the limit of quantification (LOQ) was 0.52 μg/mL and the limit of detection was 0.15 μg/mL. Intermediate precision was evaluated, expressed as inter-day variation (RSD = 1.13) and intra-day (RSD = 1.16). The accuracy obtained was the recovery of 102.87 ± 9.84, considering the exact method proposed. To evaluate the robustness, the results showed little variation, concluding that a robust method. The method was fast, robust and suitable for quantification of suspensions of COL nanoparticles and can be used to evaluate the efficiency of COL encapsulation.