## 4 Conclusions  
In this work, value-added compounds from _C. pepo_ seeds were extracted using green techniques--UAE and MAE--and an eco-friendly solvent--water, aiming a potential use as nutraceutical ingredient. Two different ratio conditions were employed.  
FRAP scavenging activity ranged from 45.80 to 71.09 \(\upmu\)mol FSE/g DW (UAE condition 2 and MAE condition 1, respectively), while DPPH values varied between 4.35 mg TE/g DW (UAE condition 1) and 5.08 mg TE/g DW (MAE condition 2). The extract with the best performance at ABTS assay was MAE condition 1 (13.29 mg AAE/g DW), while UAE condition 2 was the worst (11.38 mg AAE/g DW). The extract with the highest TPC value was MAE condition 2 (16.89 mg GAE/g DW), and UAE condition 1 had the lowest (12.17 mg GAE/g DW). All extracts displayed a remarkable scavenging capacity for HOCI, with IC\({}_{50}\) values ranging from 1.88 (UAE condition 1) to 13.50 \(\upmu\)g/mL (UAE condition 2).  
HPLC-PDA analysis revealed the presence of alkaloids, chalconoids, flavanols, flavanones, flavonols, phenolic acids and stilbenoids. A total of 21 phenolic compounds were identified, with catechin, caffeine and gallic acid being present in higher quantities. No adverse effects were detected in Caco-2 viability to MAE extracts up to 1000 \(\upmu\)g/mL, while all the other conditions tested led to a slight decrease of cell viability.  
The results shown that a higher sample-to-solvent ratio led to an increase in the amount of phenolic compounds present in both MAE and UAE extracts, which led to better results in ORAC, O\({}_{2}\)\({}^{\bullet-}\), TPC and DPPH assays. Overall, these results highlighted that _C. pepo_ seeds are a potential source of antioxidant/antiradical compounds and that the extract MAE condition 2 is rich in bioactive compounds with potential uses in the nutraceutical field.  
Moreover, the results obtained in this study show that higher sample-to-solvent ratios using MAE technology can lead to the extraction of a greater amount of phenolic compounds. Therefore, further studies are needed to screen the effects on neuronal enzymes (such as acetylcholinesterase or butylcholinesteraseterase) as well as 3D intestinal permeation studies to screen the amounts that are absorbed at intestinal level.  
Conceptualization, F.R.; methodology, C.M., A.M.S., A.S.F., M.M.M. and F.R.; software, A.M.S., C.M. and M.M.M.; validation, M.M., F.R. and C.D.-M.; formal analysis, C.M., A.M.S., M.M.M. and F.R.; investigation, C.M., A.M.S., A.S.F., M.M.M. and F.R; resources, C.D.-M. and F.R.; writing--original draft preparation, C.M. and A.M.S.; writing--review and editing, F.R., M.M.M. and C.D.-M.; supervision, F.R.; project administration, F.R.; funding acquisition, C.D.-M. and F.R. All authors have read and agreed to the published version of the manuscript.  
**Funding:** This research was funded by UIDB/50006/2020 and UIDP/50006/2020 by the Fundacao para a Ciencia e a Tecnologia (FCT)/Ministerio da Ciencia, Tecnologia e Ensino Superior (MCTES) through national funds.  
**Institutional Review Board Statement:** Not applicable.  
**Informed Consent Statement:** Not applicable.  
**Data Availability Statement:** Data available on request due to restrictions e.g., privacy or ethical.  
**Acknowledgments:** [PERSON] (SFRH/BD/144994/2019) and [PERSON] (2020-07519.BD) are thankful for the Ph.D. grants financed by POPH-QREN and subsidized by the European Science Foundation and Ministerio da Ciencia, Tecnologia e Ensino Superior. [PERSON] (CEECIND/02702/2017) and [PERSON] (CEECIND/01886/2020) are thankful for their contracts financed by FCT/MCTES--CEEC Individual Program Contract.  
**Conflicts of Interest:** The authors declare no conflict of interest.