Evaluation of PEGylation efficacy of curcumin-loaded nanoemulsions using complementary methods to assess protein interactions and physicochemical properties

Abstract

Nanoemulsions (NEs) are frequently used in the food, cosmetics and pharmaceutical industries to deliver nutraceuticals, pharmaceutical or cosmetic active ingredients. When administering NEs parenterally, various stabilisers are added to prevent rapid plasma clearance and to successfully deliver the active ingredient to the target site. For this purpose, PEGylation is often used to prolong the circulation time of the droplets. However, the problem is to determine the optimal concentration of the PEGylating agent – the PEGylation efficacy – that ensures adequate surface protection. This is a particular challenge when the active ingredient is incorporated into the stabilising layer, where any changes could disrupt the stability of the droplet. For this reason, we aimed to determine the optimal concentration of PEG2000-DSPE for surface protection of curcumin-loaded NEs for parenteral administration using electron paramagnetic resonance (EPR) spectroscopy. NEs were prepared using the high pressure homogenisation technique with 0.1 %, 0.3 % or 0.6 % of the PEGylated phospholipid. A droplet size of approximately 100 nm and polydispersity index below 0.25 indicated suitability for parenteral application. EPR analysis showed that PEG2000-DSPE had a stabilising effect on selected NEs, which was most pronounced in the part of the stabilising layer closest to the aqueous phase. To confirm these results, protein interaction studies were carried out using dynamic light scattering, UV–Vis spectroscopy, atomic force microscopy and release studies from protein-enriched media – bovine serum albumin (BSA) or foetal bovine serum (FBS) in phosphate-buffered saline. These analyses confirmed that the addition of PEG2000-DSPE reduced protein binding to the droplets as a function of concentration, with 0.3 % providing the best protection for the droplets. Our conclusions from the EPR spectroscopy study demonstrate the usefulness of EPR in determining the optimal concentrations of PEGylating agents for surface coverage and its usefulness in the formulation development phase.