## 4 Discussion
### Genotoxic Evaluation  
The observed protective effects of elderberry hydrolate against genotoxic stress, as revealed through the Comet assay with H2O2 and SN challenges, align with the known properties of elderberry, particularly its rich phytochemical composition [24]. The elevation in DNA damage seen in the 'C+' group exposed solely to H2O2 underscores the pronounced genotoxic impact of hydrogen peroxide. In contrast, the concentration-dependent relationship observed in the elderberry hydrolate-treated groups suggests a nuanced interplay between the hydrolate concentrations and DNA protection. This aligns with the determined antioxidant properties of elderberry hydrolate, as antioxidants are known to scavenge reactive oxygen species, reducing oxidative stress-induced damage to cellular components, including DNA. The optimal outcome observed with the 10% elderberry hydrolate treatment in the SN condition and the 1% treatment with the H2O2 condition suggests a potential concentration-specific protective effect. This finding may be attributed to the optimal balance of bioactive compounds at this concentration, as higher concentrations displayed trends of increased DNA damage. The 1% and 10% concentrations, with their minimal DNA damage and a notable reduction in genotoxicity, highlight the potential of elderberry hydrolate to mitigate oxidative stress. Also, the hydrolate likely enhances the cellular antioxidant defense systems, including the upregulation of antioxidant enzymes or direct scavenging of free radicals. Furthermore, the diverse outcomes observed with the different hydrolate concentrations and the challenges underscore the complexity of elderberry hydrolate's effects on DNA damage. The nuances in the results may be linked to variations in the concentrations of specific phytochemicals within elderberry hydrolate, each contributing differently to its overall protective capacity. The presence of phenylacetaldehyde, 2-acetyl-pyrrole, and the unidentified compound in elderberry hydrolate suggests potential bioactivity that could correlate with the observed genotoxic effects. phenylacetaldehyde, predominant (26.7%) in the hydrate as determined when using the unprocessed sample, as well as in both ethyl acetate (100%; it was the only detected compound) and hexane (82.6%) extracts, is a known fragrant compound that has demonstrated various biological activities, including antimicrobial and antioxidant properties [25; 26; 27]. These properties might contribute to the antigenotoxic effects observed in the Comet assay, as antioxidants are known to mitigate oxidative stress-induced DNA damage. The unidentified compound, which constitutes a significant portion of the hydrolate (59.7%; unprocessed hydrolate), warrants further investigation to elucidate its structure and potential biological activity. The genotoxic evaluation, which showed a reduction in DNA damage, indicates that the combined effect of these compounds may confer protective properties against genotoxic agents. Moreover, 2-acetyl-pyrrole, another identified compound, has been studied for its potential biological activities, including its antioxidant properties [28; 29]. This compound might also play a role in the antigenotoxic effects observed, although its exact mechanism of action in this context remains to be further explored. n-Hexanal (3.3%) and furfural (4.5%), detected in hexane extracts at lower concentrations, also possess known biological activities, including antioxidant properties [30; 31]. Their presence, although minor, might synergistically enhance the overall antigenotoxic effect of the hydrolate. (_E_)-beta-Damascenone, a compound present in hexane extracts at 2.2%, is another contributor to the biological activity of the hydrolate. Its antioxidant properties further support the hydrolate's ability to protect against DNA damage [32]. Furthermore, the 2-acetyl-pyrrole (13.7%; unprocessed hydrolate) in elderberry hydrolate may enhance its ability to protect against DNA damage.