## 2 Materials and Methods
### Determination of Polyphenols Profile by UPLC-Q-TOF-MS  
Phenolic compounds were identified using an ultra-performance liquid chromatograph equipped with a photodiode array detector, mass spectrometer, and an electrospray ionization source (UPLC-PDA-TQD-EPI, Waters, Milford, MA, USA) [31; 33]. An UPLC BEH C18 column (100 \(\times\) 2.1 mm, 1.7 \(\upmu\)m, Waters, Warsaw, Poland) was used for the separation of phe \begin{table}
\begin{tabular}{p{42.7 pt} p{42.7 pt} p{42.7 pt} p{42.7 pt} p{42.7 pt} p{42.7 pt} p{42.7 pt}} \hline \hline \multirow{2}{*}{**No.**} & \multirow{2}{*}{**Ingredients**} & \multirow{2}{*}{**Dose of Extract**} & \multicolumn{4}{c}{**Polymerizing Compounds**} & \multicolumn{1}{p{42.7 pt}}{**Encapsulation**} & \multicolumn{1}{p{42.7 pt}}{**Microcapsule Size**} \\ \cline{4-7}  & & & **Sodium Alginate (Alg)** & **Pectin (Pect)** & **Protein Isolate (WPI)** & **(\%)** & **(\(\upmu\)m)** \\ \hline A & & & + & & & 8.79 \(\pm\) 0.29 \({}^{\text{a}}\) & 607.06 \(\pm\) 48.54 \({}^{\text{c}}\) \\ B & Crude & & + & + & & 14.88 \(\pm\) 0.37 \({}^{\text{b}}\) & 609.17 \(\pm\) 35.02 \({}^{\text{c}}\) \\ C & extract (CE) & & + & & + & 9.37 \(\pm\) 0.29 \({}^{\text{a}}\) & 682.02 \(\pm\) 52.72 \({}^{\text{f}}\) \\ D & & 0.05\% & + & + & + & 17.48 \(\pm\) 0.31 \({}^{\text{c}}\) & 690.09 \(\pm\) 41.96 \({}^{\text{g}}\) \\ \hline E & & & + & & & 29.95 \(\pm\) 0.09 \({}^{\text{d}}\) & 562.91 \(\pm\) 20.07 \({}^{\text{a}}\) \\ F & Purified & & + & + & & 71.65 \(\pm\) 0.17 \({}^{\text{f}}\) & 584.85 \(\pm\) 43.53 \({}^{\text{b}}\) \\ G & extract (PE) & & + & & + & 63.35 \(\pm\) 0.09 \({}^{\text{e}}\) & 710.06 \(\pm\) 50.86 \({}^{\text{h}}\) \\ H & & & + & + & + & 74.55 \(\pm\) 0.18 \({}^{\text{g}}\) & 720.73 \(\pm\) 72.97 \({}^{\text{j}}\) \\ \hline I & & & + & & & 618.24 \(\pm\) 45.41 \({}^{\text{d}}\) \\ J & Control & & + & + & & 632.74 \(\pm\) 36.74 \({}^{\text{e}}\) \\ K & & & + & + & & 717.56 \(\pm\) 47.21 \({}^{\text{i}}\) \\ L & & & + & + & + & 721.94 \(\pm\) 62.77 \({}^{\text{j}}\) \\ \hline \hline \end{tabular} Abbreviations: +, ingredient present in the encapsulation mixture. The results for EE (n = 3) and microcapsule diameter (n = 10) were expressed as mean and SD. Statistically significant differences within a given parameter are marked with different letters (\(\text{p}<\) 0.05; Duncan’s test).  
\end{table} 
Table 1: Composition of microencapsulated blueberry extracts and encapsulation efficiency. nols, which was maintained at 50 \({}^{\circ}\)C at a flow rate of 0.35 mL/min with an injection volume of 5 \(\upmu\)L. The mobile phase consisting of solvent A (water with 1% formic acid solution) and solvent B (40% acetonitrile with 1% formic acid solution) was used for the separation of anthocyanins, while for the remaining phenolic compounds, solvent A (water) and solvent B (40% acetonitrile) were used. The data obtained from the identification were calculated using MassLynx v.4.1 software (Waters, Milford, MA, USA). The compounds were identified based on retention time, molecular weight, UV-VIS spectrum, MS/MS ions, and data in the literature. The quantification of polyphenic compounds was performed using different internal standards. The concentrations of polyphenic compounds were calculated by preparing a calibration curve of mass concentration versus peak area. The slope of the regression line and the correlation coefficient (R2) values for each standard curve were obtained using MS Excel 2019 software. The limits of detection (LOD) and limits of quantification (LOQ) were calculated for each sample in triplicate. The calibration curves were obtained sequentially by plotting the concentration versus peak area. The mean slope (S) and the standard deviation of the intercept (\(\delta\)) were calculated from the standard curve of triplicates. The LOD and LOQ were calculated using the following equations:  
LOD = 3.3 \(\times\) (6/S)  
LCOQ = 10 \(\times\) (6/S)  
The intra- and interday variations were determined using relative standard deviation (RSD) values, which were <3.5% for all the analyzed compounds.