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Published:
Jul 19, 2021Keywords:
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Main Article Content
Simple and sensitive chromatographic method development for in-vitro and in-vivo analysis of doxorubicin-loaded poly lactic-co-glycolic acid nanoparticles
Abstract
Purpose: To develop a bioanalytical high performance liquid chromatography (HPLC) method for the quantification of doxorubicin in biological fluids and polymeric nano-formulations.
Methods: Analysis of doxorubicin in polymeric nanoparticles and biological samples was carried out at 252 nm using Purospher® RP-18 end-capped column (250mm×4.6mm,5µm) secured with a guard column cartridge RP18 (30mm×4.6mm,10µm). The mobile phase used was 0.025M phosphate buffer and acetonitrile (ACN) (65:35, v:v) in isocratic mode at a flow rate of 0.9 ml/min, run time of 10 min, column oven temperature of 30 °C, and injection volume of 40 μL.
Results: The standard curve for doxorubicin was linear (0.999) in the concentration range of 0.022 - 1.00 μg/mL in human and albino mice plasma. Nominal retention times of doxorubicin and IS were 3.5 and 5.5 min, respectively. Mean recovery was within acceptable limits (100 ± 2 %). The limit of detection (LOD) and limit of quantification (LOQ) were 0.012 and 0.022 μg/mL, respectively.
Conclusion: A reliable HPLC method has been successfully developed, validated and applied for the in vitro analysis of doxorubicin released from polymeric nanoparticles and in vivo pharmacokinetic studies in albino mice. The method may also be applicable to the analysis of doxorubicin in human fluids.
