## 3 Results
### Impact of the Solvent and Extraction Method on the Recovery of Phytochemicals
Besides the phenolic compounds, terpenoids were also found to possess a high discrimination ability. Particularly, we found triterpenes (lupine, 7-oxo-10 alpha-cucurbitadienol acetate) and sesquiterpene lactones (i.e., vernoflexin and crediasiide E) with VIP scores ranging from 1.32 to 1.36. The best extraction solvents for these terpenoid biomarkers were EtOH (sum LogFC = 33.88) and EA (sum LogFC = 30.87; Table S2). Figure 2: The number, cumulation in classes of compounds, and differences among three methods of extraction represented using variables importance in projection (VIP) markers carried out from three orthogonal projections to latent structures discriminant analysis (OPLS-DA) models such as homogenizer-assisted extraction (HAE), maceration (MAC), and Soxhet (SOX). (**A**) A cumulative graph of the most representative VIP markers (VIP score > 1.2) resulting from the three OPLS-DA models, according to their compound’s classification. The number reported in the bar corresponds to the number of VIP markers belonging to compounds classes, while the bar color represents the different OPLS-DA models: HAE (blue), MAC (yellow), and SOX (green). (**B**) Venn analysis of three lists of VIP markers resulting from OPLS-DA models. Regarding MAC, we detected 44 discriminant compounds (Table S3). This method was not very effective in extracting BCs, resulting in a lower average number in many classes of compounds compared to HAE and SOX (Figure 2A). The OPLS-DA model allowed detecting 25 polyphenols, reporting flavonoids (dihydroquerectin, luteolin, malvidin 3-O-(6\"-acetyl-galactoside), kaempfer 7-O-glucoside), phenolic acids (caffeic acid 4-O-glucose, 4-hydroxybenzoic acid 4-O-glucose, p-coumarooyl glucose), lignans (isolariciresinol, medioresinol), and other polyphenols (4-hydroxycournarin, 5-heneicosylrescornol) as the most discriminant among the different solvents used. Moreover, different sesquiterpene lactones, such as dehydrotocostuslactone, iexerin-E or -X, and 10-epi-8-deoxycumambarin were included among the most discriminant compounds for the methanolic extracts (Table S3). Concerning the SOX extraction method (Table S4), the sesquiterpene lactones were highly discriminant, with some compounds having the highest VIP score, such as 1\(\beta\),6\(\alpha\)-dihydroxycostic acid, ferolide, 11-epiartesin, and xanthumin. Additionally, two alkaloids, namely, trigamine and schelhammeridine, were particularly abundant in the ethanolic extracts. Regarding polyphenols, we found kaempfer 3-O-(6\"-acetyl-galactoside) 7-O-hamnoside (flavonols), coumestrol (LMWPs), and dihydrocaffeic acid (phenolic acids). Afterward, a Venn analysis was performed using VIP markers obtained from the three OPLS-DA models (i.e., for each method of extraction) in order to investigate which method was best for providing efficient extraction of BCs in _R. angustifolius_ extracts, considering the same extraction solvents (Figure 2B). The three extraction methods considered were shown to share only 2.6% of common BCs, suggesting that the effect of different methods has a greater impact on the isolation of different BCs. The HAE and SOX extraction methods resulted in having the most common isolated BCs (12%) compared to MAC (7.7%). Moreover, HAE was shown to have the highest isolation capacity of the most discriminating unique BCs in _R. angustifolius_ extracts (27.4%), compared to SOX and MAC by 23.1% and 22.2%, respectively.