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- Ultrasonic-assisted extraction and natural deep eutectic solvents combination: A green strategy to improve the recovery of phenolic compounds from Lavandula Pedunculata subsp. Lusitanica (Chaytor) FrancoPublication . Mansinhos, Inês; Gonçalves, Sandra; Rodríguez Solana, Raquel; Ordóñez-Díaz, José Luis; Moreno-Rojas, José Manuel; Romano, A.The present study aimed at evaluating the effectiveness of different natural deep eutectic solvents (NADES) on the extraction of phenolic compounds from Lavandula pedunculata subsp. lusitanica (Chaytor) Franco, on the antioxidant activity, and acetylcholinesterase (AChE), butyrylcholinesterase (BChE) and tyrosinase (Tyr) inhibitory capacities. Ten different NADES were used in this research and compared with conventional solvents. Ultrasound-assisted extraction (UAE) for 60 min proved to be the best extraction condition, and proline:lactic acid (1:1) and choline chloride:urea (1:2) extracts showed the highest total phenolic contents (56.00 ± 0.77 mgGAE/gdw) and antioxidant activity [64.35 ± 1.74 mgTE/gdw and 72.13 ± 0.97 mgTE/gdw in 2.2-diphenyl-1- picrylhydrazyl (DPPH) and 2.20 -azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) methods, respectively]. These extracts also exhibited enzymes inhibitory capacity particularly against Tyr and AChE. Even so, organic acid-based NADES showed to be the best extractants producing extracts with considerable ability to inhibit enzymes. Twenty-four phenolic compounds were identified by HPLC-HRMS, being rosmarinic acid, ferulic acid and salvianolic acid B the major compounds. The results confirmed that the combination of UAE and NADES provide an excellent alternative to organic solvents for sustainable and green extraction, and have huge potential for use in industrial applications involving the extraction of bioactive compounds from plants.
- Impact of temperature on Phenolic and Osmolyte contents in In Vitro cultures and micropropagated plants of two mediterranean plant species, Lavandula viridis and Thymus lotocephalusPublication . Mansinhos, Inês; Gonçalves, Sandra; Rodríguez Solana, Raquel; Ordóñez-Díaz, José Luis; Moreno-Rojas, José Manuel; Romano, AnabelaIn this study, in vitro cultures and micropropagated plants of two Mediterranean aromatic plants, Lavandula viridis L’Hér and Thymus lotocephalus López and Morales, were exposed to different temperatures (15, 20, 25, and 30 ◦C). The effect of temperature on the levels of hydrogen peroxide (H2O2 ), lipid peroxidation, and osmoprotectants (proline, soluble sugars, and soluble proteins), as well as on the phenolic profile by HPLC-HRMS and intermediates of the secondary metabolism (phenylalanine ammonia lyase (PAL) activity and shikimic acid content), was investigated. Moreover, the antioxidant activity of the plant extracts was also analyzed. Overall, considering the lipid peroxidation and H2O2 content, the extreme temperatures (15 and 30 ◦C) caused the greatest damage to both species, but the osmoprotectant response was different depending on the species and plant material. In both species, phenolic compounds and related antioxidant activity increased with the rise in temperature in the micropropagated plants, while the opposite occurred in in vitro cultures. L. viridis cultures showed the highest biosynthesis of rosmarinic acid (92.6 g/kgDW) at 15 ◦C and seem to be a good alternative to produce this valuable compound. We conclude that contrasting temperatures greatly influence both species’ primary and secondary metabolism, but the response is different depending on the plant micropropagation stage.