## 3 Results and Discussion
### Changes in Raw Material Caused by Pre-Treatment in Ethanol Solution with and without Ultrasound
The use of ultrasonic waves with a frequency of 21 (EtOH+US21) and 40 kHz (EtOH+US40) further increased the extractivityof carotenoids. During the 60 s of treatment at 21 kHz, the carotenoid content was higher by 32%, and for the frequency of 40 kHz, by 44% compared to the raw material. In the case of a treatment lasting 180 s, the carotenoid content was higher, in comparison to fresh carrot, by 104% (EOH+US21) and 144% (EOH+US40). However, the two-factor ANOVA did not confirm the significance of both factors (\(p>0.05\), Figure 2). A similar increase in the content of active compounds as a result of sonication was obtained by [PERSON] et al. [47] in the case of apple tissue. The polyphenol content after sonication at the frequency of 21 and 40 kHz was from 27 to 145% higher in comparison to the raw tissue. The increase of polyphenol content was observed with increasing the processing time and the frequency from 21 to 40 kHz. On the basis of the obtained results, it can be concluded that the use of immersion in ethyl alcohol changed the structure [11; 18] and increased the extractivity of carotenoid from carrot tissue, especially with ultrasound application and the prolongation of the processing time. \begin{table}
\begin{tabular}{c c c c c c} \hline \hline
**Type of** & **L\({}^{\star}\) (\(\star\))** & **a\({}^{\star}\) (\(\star\))** & **b\({}^{\star}\) (\(\star\))** & **h (\({}^{\circ}\))** & **\(\Delta\)E (\(\star\))** \\
**Pre-Treatment** & & & & & \\ \hline untreated & 40.6 \(\pm\) 2.1 bcd & 12.3 \(\pm\) 3.0 ab & 23.4 \(\pm\) 3.8 ab & 62.6 \(\pm\) 2.2 ef & - \\
5 s EtOH & 41.5 \(\pm\) 2.6 abcd & 14.5 \(\pm\) 3.2 a & 26.0 \(\pm\) 3.7 a & 61.1 \(\pm\) 2.7 e & 5.8 \(\pm\) 2.8 bc \\
15 s EtOH & 38.2 \(\pm\) 2.6 de & 7.5 \(\pm\) 2.6 ef & 17.2 \(\pm\) 2.4 ef & 67.2 \(\pm\) 4.6 bc & 8.7 \(\pm\) 3.3 bc \\
60 s EtOH & 38.5 \(\pm\) 2.2 de & 8.4 \(\pm\) 2.4 de & 18.8 \(\pm\) 3.4 cde & 66.3 \(\pm\) 3.1 bcd & 6.6 \(\pm\) 4.3 bc \\
180 s EtOH & 41.9 \(\pm\) 3.4 abc & 9.8 \(\pm\) 2.3 bcd & 18.9 \(\pm\) 3.0 cde & 62.8 \(\pm\) 2.3 ef & 6.6 \(\pm\) 3.0 bc \\
5 s EtOH+US21 & 41.1 \(\pm\) 3.5 bcd & 9.9 \(\pm\) 2.5 bcd & 19.0 \(\pm\) 3.3 cde & 62.9 \(\pm\) 2.5 ef & 6.7 \(\pm\) 3.0 bc \\
15 s EtOH+US21 & 40.1 \(\pm\) 3.9 bcd & 8.4 \(\pm\) 3.9 de & 17.2 \(\pm\) 4.2 ef & 65.5 \(\pm\) 3.7 bcde & 9.4 \(\pm\) 4.0 ab \\
60 s EtOH+US21 & 41.9 \(\pm\) 3.9 abc & 9.5 \(\pm\) 4.0 bcd & 19.4 \(\pm\) 3.2 cde & 65.4 \(\pm\) 4.2 bcde & 6.8 \(\pm\) 2.8 bc \\
180 s EtOH+US21 & 43.2 \(\pm\) 3.7 ab & 7.3 \(\pm\) 2.3 ef & 18.0 \(\pm\) 3.1 def & 68.4 \(\pm\) 3.7 b & 8.9 \(\pm\) 2.9 bc \\
5 s EtOH + US40 & 37.1 \(\pm\) 1.9 e & 4.5 \(\pm\) 2.2 f & 14.8 \(\pm\) 2.8 f & 74.0 \(\pm\) 4.8 a & 12.3 \(\pm\) 3.5 a \\
15 s EtOH + US40 & 38.9 \(\pm\) 2.3 cde & 8.4 \(\pm\) 3.3 de & 18.1 \(\pm\) 3.7 cdef & 65.8 \(\pm\) 3.7 bcde & 7.3 \(\pm\) 4.2 bc \\
60 s EtOH + US40 & 43.0 \(\pm\) 3.9 ab & 11.4 \(\pm\) 3.2 abc & 21.5 \(\pm\) 3.7 bcd & 61.8 \(\pm\) 3.0 ef & 5.4 \(\pm\) 2.8 c \\
180 s EtOH + US40 & 44.7 \(\pm\) 4.0 a & 11.3 \(\pm\) 3.5 bc & 21.8 \(\pm\) 3.4 bc & 63.1 \(\pm\) 3.7 def & 7.1 \(\pm\) 2.3 bc \\ \hline \hline \end{tabular}  
* - a, b, etc.—different letters in each column indicate different homogeneous groups (one-way ANOVA, \(\alpha=0.05\)). The results of two-way ANOVA indicate the significance (\({}^{\star}\)) or insignificance (\(\cdot\)) of the influence of the type and time of treatment (\(\alpha=0.05\)).  
\end{table} 
Table 2: Colour parameters (L\({}^{\star}\), a\({}^{\star}\), b\({}^{\star}\)), hue angle (h\({}^{\circ}\)) and total colour difference (\(\Delta\)E, in comparison to raw material) of carrot tissue pre-treated without and with ultrasound; error bars indicate \(\pm\) SD (standard deviation calculated from at least 10 repetitions).