# Metabolic Profiling of _Conyza sumatrensis_ ([PERSON].) [PERSON] from Lugazi, Uganda
## 3 Results and Discussion
### Compound Prediction with In Silico Techniques and Database Matches

Table 3: Library matches with the corresponding name, database, and URL. Cluster 4 has been classified as acetylated polyamines. Natural polyamines such as N-acetylspermine or spermidine can play a major role in plant stress tolerance. They can form conjugates with cinnamic acid derivatives, serve as reserve polyamines for proliferative tissue, and act against pathogens or for the detoxification of phenolic compounds [42]. Polyamines are also used as polymers in a variety of ways, e.g., as a carrier for drug delivery systems, as a polyethylene glycol alternative, and as a food additive [43]. Through accumulation in the food chain and via wastewater, these polymers can enter the environment and then be taken up by plants [44]. In the environment, they can have the same effect on plants as nitrogenous fertilizers [45]. Cluster 5 comprises three nodes representing flavonoids with spectral matches supporting this finding. Cluster 6 is also classified as flavonoids with one node assigned as a phenylpropanol related Mass2 Motif (rhabdus_motif_91) by MS2 LDA. Overall, with MS2 LDA, some molecular class predictions were consistent with those of the molecular network and spectral library hits. Flavonoids are secondary metabolites with a polyphenic structure and are ubiquitous in plants [46]. They are, e.g., responsible for the color, smell, and taste of plants and have several important physiological functions, such as regulating cell growth, regulating UV filters, scavenging reactive oxygen species, and attracting pollinators [27]. Flavonoids are produced by plants against microbial infestation, among other conditions, and have been shown to have positive effects on human health. The positive impact is explained by antioxidant properties, which are mediated by substituted hydroxyl groups that can scavenge free radicals. Studies have also shown effects against cardiovascular diseases and cancer, as well as hepatocyte, antibacterial, antiviral, and anti-inflammatory effects [47]. All signals in cluster 5 were only present in the aqueous extracts, and since flavonoids as aglycons are usually non-polar and poorly water-soluble, it is likely that these putative flavonoids would be glycosylated, improving water solubility [48]. And indeed, the library match for (ID 55 within this cluster) showed a glycosylated apigenin derivative. Cluster 7 showed a spectral match to a structure of a benzoic acid methyl ester called galutherin that is present in both ethanolic and water extracts. This compound is of scientific interest due to its analgesic and anti-inflammatory effects similar to those of salicylic acid, which was shown by [PERSON] et al. [49]. Notably, galutherin has mainly been found in plants of the genus heather family (Ericaceae). Further studies are thus necessary to evaluate the presence of galutherin in _C. sumatrensis_ and the possibility of analgesic properties of this plant. The use of in silico tools for compound class prediction has become increasingly important and can support classical library and database searches [18]. However, it is essential to note the limitations of our approach: although we were able to predict natural product classes, we were mainly unable to assign the features to specific compounds. The purification and structural characterization of these compounds are essential to facilitate their availability for bioassays, enabling a more comprehensive exploration of the plant's bioactivity profile. Notably, more than half of the nodes could not be predicted using these tools. Similar outcomes in other studies have been explained by poor ionization, fragmentation, and separation of analytes during the LC-MS/MS run [18; 19]. Overall, this study demonstrates the potency and limitations of a multi-technique analytical approach in the field of plant metabolomics. Database searches and comparisons of the literature enabled the preliminary identification of over 40 substances present in the plant that have not been reported before from _C. sumatrensis_.