## 3 Results  
### pH Assay, Total Extract, and Total Acidity  
To obtain spirit drinks of the highest quality, it is necessary to control their physico-chemical characteristics. Several analytical determinations, namely pH, total acidity, and total extract were done to characterize prepared macerates. The results show that the pH of mistletoe macerates varies between 5.5 and 6.25 (Figure 1), with the lowest pH measured in sample A_20 and the highest in sample D_40. It can be observed that the composition of the water-alcohol base significantly affects the differences in the pH of the macerates as a function of plant concentration. Indeed, the pH values of the aqueous alcohol so lutions themselves differ significantly, with a higher proportion of ethanol leading to a neutral medium and a higher proportion of water leading to an acidic medium. Interestingly, the addition of plants to water-alcohol bases with lower ethanol concentrations (25% and 40% \(v/v\)) increases the pH, while the addition of plants to water-alcohol bases with higher ethanol concentrations (55% and 70% \(v/v\)) decreases the pH. This indicates that the maceration medium significantly affects the type and amount of compounds to be extracted. The total extract is shown in Figure 2. As expected, the amount of extracted substances in the macerate (sugars, minerals, and other organic substances) increases with increasing plant content. Therefore, macerates with a higher plant content also have a greater amount of total extract. The results show that the highest amount of total extract is obtained for macerates with 25% volume and the highest plant concentration. At the highest plant concentration, the amount of total extract decreases as the ethanol concentration increases. The total extract at the lowest plant concentration does not depend on the alcohol content. At the lowest plant concentration, the concentration gradient is the largest, so with longer maceration time (28 days), mistletoe components are extracted regardless of the alcohol base. At a medium plant concentration, the sample with 40% ethanol had the highest concentration of extracted components. Acid concentrations in the samples varied between 480 and 2880 mg/L (Figure 3). Sample B_20 had the lowest value and sample D_80 had the highest. As the amount of herbs increases, the total acidity of the macerate increases. The increase in the total acidity of the macerate with the addition of herbs shows the extraction of acids from the plants into the water-alcohol base. At low ethanol concentrations (25% \(v/v\)), extraction of polar compounds occurs due to the predominant polar nature of water. However, increasing the ethanol concentration in the macerates changes the polarity of the solvent and allows extraction of less polar compounds, which increase the concentration of total acids [18]. Mistletoe contains triterpene acids such as oleanolic, betulinic, and unsolic acids in its leaves and stem [19]. It is very rich in saturated and unsaturated fatty acids. It contains palmitic acid, arachidonic, lignoercic, behenic, and cerotic acids from the saturated acids and oleic acid, linoleic acid, and linolenic acid from the unsaturated acids [7]. Figure 1: pH values of ethanol–water solution and mistletoe macerates prepared by maceration of 20 g/L, 40 g/L, and 80 g/L mistletoe in ethanol–water solution (A: 25%, B: 40%, C: 55%, and D: 70% \(v/v\) ethanol) at room temperature and in darkness, after 28 days of maceration. Different letters indicate significant differences between samples (\(p<0.05\)). Figure 3: Total acidity of mistletoe maccrates prepared by maceration of 20 g/L, 40 g/L, and 80 g/L, mistletoe in ethanol-water solution (A: 25%, B: 40%, C: 55%, and D: 70% \(v/v\) ethanol) at room temperature and in darkness, after 28 days of maceration.