## 3 Results and Discussion
### Nanoparticle Characterization  
After dialysis and freeze-drying, the AuNPs were obtained as black powder and could be easily re-dispersed in water without flocculation. The obtainment of decorated AuNPs with the synthesized ligands was preliminary assessed with infrared spectroscopy. In particular, FT-IR/ATR analyses on solid samples of AuNPs 1-4 were performed (Figure 6). The registered IR spectra on AuNP1-4 showed the presence of several bands associated with the functional groups of the decorating ligands, for example the stretching bands of sugar OH groups around 3300 cm\({}^{-1}\), the stretching bands of CH\({}_{2}\) of pentenyl chains at 2900 cm\({}^{-1}\), the C-O stretching band of ethylene glycol chains and of sugar rings in the region of 1015 cm\({}^{-1}\), as well as bands around 1650 cm\({}^{-1}\) (stretching of aromatics) and 1350 cm\({}^{-1}\) (OH bending). The IR spectra of AuNP3 and AuNP4 present a narrow band around 830 cm\({}^{-1}\) that can be associated with the bending vibrations of the amino group of the glutamine derivative grafted on AuNP 3 and 4. Figure 5: Structure of the four multivalent nanoparticles prepared in this work. Morphological characterization of AuNP1-4 was carried out by TEM (transmission electron microscopy) analysis. The measurement of the AuNPs' size and their distribution was accomplished using _Image_) (version 1.54 bundled 64-bit) and _Origin_ (version Pro 2023 64-bit SR1) software, adopting the procedure and the processing steps described recently by [PERSON] and [PERSON] [74]. Except for the AuNP4 sample, for which we counted a little bit less than 100 nanoparticles, in AuNP1, 2 and 3, we counted more than 100 nanoparticles. The four analyzed dispersions revealed the presence of nanoparticles ranging between 2 nm and 6 nm in diameter (gold core), with those of 2 nm being the most representative one, as indicated in the diameter distribution histograms of Table 1.  
In order to further confirm the mean size of the AuNPs, the AuNP2 sample was chosen as a model and was subjected to UV-VIS spectroscopy analysis (Figure 7). The nanoparticles were dissolved in water at two different dilutions (0.5 mg/mL and 0.25 mg/mL), and only at the higher concentration was it possible to notice a weak plasmon absorption band around 520 nm, which is correlated to the presence of few nanoparticles with a mean bigger core diameter. According to the obtained results, AuNP2 and AuNP3 showed the best features in terms of final mass yield and size distribution. Moreover, AuNP2 and AuNP3 were the most representative model nanoparticles among the prepared ones, since they exposed the C-glucoside derivative **2** (AuNP2) or the L-glutamine derivative 4 (AuNP3), the two active anti-inflammatory ligands of interest of this work, and the D-glucose **Glc5 SH** derivative, used for the modulation of the dispersibility of the nanoparticles. \begin{table}
\begin{tabular}{c c c} \hline \hline
**Nanoparticle** & **TEM Micrograph** & **Diameter Distribution Histogram** \\ \hline \hline \end{tabular}
\end{table} 
Table 1: TEM micrographs and size (diameter) distribution of the prepared nanoparticles. Figure 6: FT-IR/ATR spectra of prepared AuNPs 1–4. Some diagnostic stretching and bending vibrations are circled. Individual IR spectra of AuNP1–4 are reported in the Supporting Information. For these reasons, we decided to deepen the physical-chemical characterization of AuNP2-3 by means of \({}^{1}\)H-NMR analysis, which allowed us to obtain more detailed structural information about the loading of the ligands on the AuNPs. The \({}^{1}\)H-NMR experiments were performed by dissolving the powder comprised of AuNP2 and AuNP3 in deuterium oxide; the obtained spectra show broad signals of the ligands, typical for this kind of nanosystem [64, 66]. In particular, signals of D-glucose derivative **Glc5 SH**, C-glucose derivative **2** (in AuNP2) and L-glutamine derivative **4** (in AuNP3) grafted onto a nanosized entity are clearly visible in the respective \({}^{1}\)H-NMR spectra (Figure 8A).