Citation: Lut , ˘ a, E.A.; Bit , ˘ a, A.; Moros , an, A.; Mihaiescu, D.E.; Ghica, M.; Mihai, D.P.; Olaru, O.T.; Deculescu-Ionit , ˘ a, T.; Dut , u, L.E.; Popescu, M.L.; et al. The Influence of Phytosociological Cultivation and Fertilization on Polyphenolic Content of Menthae and Melissae folium and Evaluation of Antioxidant Properties through In Vitro and In Silico Methods. Plants 2022, 11, 2398. https://doi.org/10.3390/ plants11182398 Academic Editor: Ahmed A. Hussein Received: 24 August 2022 Accepted: 13 September 2022 Published: 14 September 2022 Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affil- iations. Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). plants Article The Influence of Phytosociological Cultivation and Fertilization on Polyphenolic Content of Menthae and Melissae folium and Evaluation of Antioxidant Properties through In Vitro and In Silico Methods Emanuela Alice Lut , ă 1, * ,† , Andrei Bit , ă 2,† , Alina Moros , an 3, * , Dan Eduard Mihaiescu 3 , Manuela Ghica 1,† , Dragos , Paul Mihai 1, * ,† , Octavian Tudorel Olaru 1, * ,† , Teodora Deculescu-Ionit , ă 1 , Ligia Elena Dut , u 1 , Maria Lidia Popescu 1 , Liliana Costea 1 , George Mihai Nitulescu 1 , Dumitru Lupuliasa 1,‡ , Rica Boscencu 1,‡ and Cerasela Elena Gîrd 1,‡ 1 Faculty of Pharmacy, University of Medicine and Pharmacy “Carol Davila”, Traian Vuia 6, 020956 Bucharest, Romania 2 Department of Pharmacognosy & Phytotherapy, Faculty of Pharmacy, University of Medicine and Pharmacy of Craiova, Petru Rares , 2, 200349 Craiova, Romania 3 Department of Organic Chemistry “Costin Nenit , escu”, Faculty of Chemical Engineering and Biotechnologies, University of Politehnica, Gheorghe Polizu 1-7, 011061 Bucharest, Romania * Correspondence: emanuela.luta@drd.umfcd.ro (E.A.L.); alina.morosan@upb.ro (A.M.); dragos_mihai@umfcd.ro (D.P.M.); octavian.olaru@umfcd.ro (O.T.O.) † These authors contributed equally to this work. ‡ These authors contributed equally to this work. Abstract: Since medicinal plants are widely used in treating various diseases, phytoconstituents enrichment strategies are of high interest for plant growers. First of all, we investigated the im- pact of phytosociological cultivation on polyphenolic content (total flavonoids—TFL, and total polyphenols—TPC) of peppermint (Mentha piperita L.) and lemon balm (Melissa officinalis L.) leaves, using spectrophotometric methods. Secondly, the influence of chemical (NPK) and organic (BIO) fertilization on polyphenolic content and plant material quality was also assessed. Dry extracts were obtained from harvested leaves using hydroethanolic extraction solvents for further qualitative and quantitative assessment of phytoconstituents by FT-ICR MS and UHPLC-MS. Furthermore, the antioxidant activity of leaf extracts was determined in vitro using DPPH, ABTS and FRAP meth- ods. Molecular docking simulations were employed to further evaluate the antioxidant potential of obtained extracts, predicting the interactions of identified phytochemicals with sirtuins. The concentration of polyphenols was higher in the plant material harvested from the phytosociological culture. Moreover, the use of BIO fertilizer led to the biosynthesis of a higher content of polyphenols. Higher amounts of phytochemicals, such as caffeic acid, were determined in extracts obtained from phytosociological crops. The antioxidant activity was dependent on polyphenols concentration, more potent inhibition values being observed for the extracts obtained from the phytosociological batches. Molecular docking studies and MM/PBSA calculations revealed that the obtained extracts have the potential to directly activate sirtuins 1, 5 and 6 through several polyphenolic compounds, such as rosmarinic acid, thus complementing the free radical scavenging activity with the potential stimulation of endogenous antioxidant defense mechanisms. In conclusion, growing medicinal plants in phytosociological cultures treated with biofertilizers can have a positive impact on plant material quality, concentration in active constituents and biological activity. Keywords: Mentha piperita L. leaves extract; Melissa officinalis L. leaves extract; phytosociology; polyphenolic content; FT-ICR MS; UHPLC-MS; antioxidant activity; molecular docking; sirtuins Plants 2022, 11, 2398. https://doi.org/10.3390/plants11182398 https://www.mdpi.com/journal/plants