Asymmetric flow field-flow fractionation (AF4) with fluorescence and multi-detector analysis for direct, real-time, size-resolved measurements of drug release from polymeric nanoparticles
Polymeric nanoparticles (NPs) are typically designed to enhance the efficiency of drug delivery by controlling the drug release rate. Hence, it is critical to obtain an accurate drug release profile. This study presents the first application of asymmetric flow field-flow fractionation (AF4) with fluorescence detection (FLD) to quantify release profiles of fluorescent drugs from polymeric NPs, specifically poly(lactic-co-glycolic acid) NPs loaded with enrofloxacin (PLGA-Enro NPs). In contrast to conventional measurements requiring separation of the NPs and dissolved drugs (typically by dialysis) prior to quantification, AF4 provides in situ removal of unincorporated drugs, while the judicious combination of online FLD and UV detection selectively provides the entrapped drug and PLGA NP concentrations, respectively, and hence the drug loading. NP size and shape factors are simultaneously obtained by online dynamic and multi-angle light scattering (DLS, MALS) detectors. The AF4 and dialysis approaches were compared to evaluate drug release from PLGA-Enro NPs containing a high proportion (≈ 94%) of unincorporated (burst release) drug at three temperatures spanning the glass transition temperature (T ≈ 33 °C) of the NPs. The AF4 method clearly captured the temperature dependence of the drug release relative to T (from no release at 20 °C to rapid release at 37 °C). In contrast, dialysis was not able to distinguish differences in the extent or rate of release of the entrapped drug because of interferences from the burst release, as well as the dialysis lag time, as supported through a diffusion model and validation experiments on purified NPs with low burst release. Finally, the multi-detector AF4 analysis yielded unique size-dependent release profiles across the entire NP size distribution, with smaller NPs showing faster release consistent with radial diffusion from the NPs. Overall, this study demonstrates the novel application and advantages of multi-detector AF4 methods, particularly AF4-FLD, to obtain direct, size-resolved release profiles of fluorescent drugs from polymeric NPs.
Publication Source (Journal or Book title)
Journal of controlled release : official journal of the Controlled Release Society
Shakiba, S., Astete, C. E., Cueto, R., Rodrigues, D. F., Sabliov, C. M., & Louie, S. M. (2021). Asymmetric flow field-flow fractionation (AF4) with fluorescence and multi-detector analysis for direct, real-time, size-resolved measurements of drug release from polymeric nanoparticles. Journal of controlled release : official journal of the Controlled Release Society, 338, 410-421. https://doi.org/10.1016/j.jconrel.2021.08.041