beech wood sawdust was isolated as described elsewhere.26 (link) Lignin nanoparticles with entrapped essential
oils (EO-LNPs) were prepared based on the protocols presented in Zikeli
et al.23 (link),26 (link) with the following adaptations: After dissolving
300 mg of AL in 10 mL of DMSO, 150 μL of the respective pure
EOs was added and further mixed for 5 min. After filtering through
0.45 μm CHROMAFIL Xtra syringe filters (Macherey-Nagel GmbH
& Co. KG, Düren, Germany), the solutions were filled in
dialysis bags (SpectraPor 1 Dialysis Membrane Standard RC Tubing,
6–8 kDa, Spectrum Labs, USA) and dialyzed against 4 L of distilled
water for 1 h. A short dialysis time was chosen to prevent a concomitant
release of already entrapped EOs and thus study the very first phase
of EO-LNP formation. The resulting EO-LNP dispersions were stored
in the fridge until further analysis. Aliquots of the EO-LNP dispersions
were freeze-dried for further analysis as well as for in vitro release
experiments.