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

Luciane Stankiewski, Ana Cristina Oltramari Toledo, Melissa Marques Gonçalves, Barbara Justus, Traudi Klein, Josiane de Fátima Padilha Paula


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.


Cholecalciferol, nanoparticles, Vitamin D3

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