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  • Influence of elaboration process on chemical, biological, and sensory characteristics of European pennyroyal liqueurs
    Publication . Rodríguez Solana, Raquel; Esteves, Eduardo; Mansinhos, Inês; Gonçalves, Sandra; Perez-Santin, Efren; Galego, Ludovina; Romano, Anabela
    BACKGROUND: Mentha pulegium is an aromatic and medicinal plant used in different Mediterranean herb-based liqueurs, whose phytochemical composition depends on the elaboration technique used. The aim of this work was to investigate differences in the sensory, chemical, and biological characteristics of pennyroyal liqueurs elaborated by hydro-alcoholic (HI) and aqueous (AI) infusions, percolation (P), maceration (M), and distillation (D) techniques, and by the mixture of spirit with pennyroyal essential oil (EO). The volatile composition determined by gas chromatography-mass spectrometry (GC-MS) and gas chromatography coupled to flame ionization detection (GC-FID), the total phenolic (TPC) and flavonoid (TFC) contents, the antioxidant (AC), and enzyme (alpha-glucosidase, tyrosinase, and acetylcholinesterase) inhibitory capacities, and the sensory analysis by napping, were evaluated in the liqueurs obtained. RESULTS: Monoterpenes and sesquiterpenes were the main volatiles present in liqueurs. The M, P, HI, and AI liqueurs showed the highest TPC, TFC, and AC values; and pulegone, the main hepatotoxic chemotype, was found in concentrations that gave rise to concern. The D and EO liqueurs stand out for their acceptable pulegone content and greater tyrosinase inhibition. Similar liqueur classifications were achieved by sensory analyses, and physico-chemical and biological analyses using multiple factor and principal component analyses, respectively. CONCLUSION: These results are an important contribution to the identification of the most appropriate elaboration conditions for herbal liqueurs with the healthiest chemical composition and functional properties. (c) 2020 Society of Chemical Industry
  • Elicitation improves rosmarinic acid content and antioxidant activity in Thymus lotocephalus shoot cultures
    Publication . Gonçalves, Sandra; Mansinhos, Inês; Rodríguez Solana, Raquel; Perez-Santin, Efren; Coelho, Natacha; Romano, Anabela
    Thymus lotocephalus G. Lopez & R. Morales is an aromatic species endemic to the south of Portugal with medicinal properties. The aim of this work was to evaluate the effect of elicitors, cytokinin, and different sucrose concentrations on chemical composition and antioxidant activity of extracts from in vitro regenerated shoots. Elicitors (YE: yeast extract, salicylic acid, and AgNO3) were added directly to the medium. Phytochemical analysis include evaluation of total phenolic (TPC), flavonoid (TFC), and rosmarinic acid (RAC) contents. The TPCs and TFCs were determined by Folin-Ciocalteu and aluminum chloride calorimetric assays, respectively. Rosmarinic acid (RA), the main compound found in the studied extracts, was quantified by liquid chromatography-photodiode array detector. Results showed that the cytokinin benzyladenine has an inhibitory effect on TPC, TFC and RAC, as well as on antioxidant activity. TPC was only significantly improved by the presence of YE (from 95.49 +/- 3.34 to 118.34 +/- 6.24 mg(GAE)/g(extract)) and TFCs were not affected by elicitation. The increase in sucrose concentration, from 2 to 4%, and the three elicitors tested, particularly YE, enhanced RACs (from 48.61 +/- 0.09 in the control medium to 78.57 +/- 0.99 mg/g(extract) in YE-elicited medium). In accordance with these results, the antioxidant activity measured by 2,2-diphenyl-1-picrylhydrazyl, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), and ferric reducing antioxidant power assays was significantly greater in extracts from elicited shoots. Overall, the results obtained suggest that shoots of T. lotocephalus are a good source of antioxidant compounds and showed that the production of RA can be promoted by altering in vitro culture conditions.
  • The influence of in vitro gastrointestinal digestion on the chemical composition and antioxidant and enzyme inhibitory capacities of carob liqueurs obtained with different elaboration techniques
    Publication . Rodríguez Solana, Raquel; Coelho, Natacha; Santos-Rufo, Antonio; Gonçalves, Sandra; Pérez-Santín, Efrén; Romano, Anabela
    Carob liqueur is a traditional Mediterranean alcoholic beverage obtained via a wide range of production techniques contributing to the different organoleptic attributes of the final product. The aim of this research was to evaluate the stability of the chemical composition and biological capacities (antioxidant and enzyme inhibition) under in vitro simulated gastrointestinal digestion of liqueurs prepared by flavouring the fig spirit with carob pulp by maceration, distillation, percolation, or aqueous and hydro-alcoholic infusions. For this purpose, the phenolic and furanic compositions, the total phenolic (TPC) and flavonoid (TFC) contents, antioxidant capacity (AC), and enzyme inhibitory potential against acethylcholinesterase, tyrosinase, α-glucosidase and α-amylase enzymes were evaluated. The content of gallic acid decreased after gastrointestinal digestion, while TPC, TFC, and AC significantly increased after each digestion phase. Overall, no significantly different enzyme inhibitions (p < 0.05) were observed among digested liqueurs, with moderate inhibition against acethylcholinesterase and tyrosinase (enzymes related with neurodegenerative diseases), and potent and low inhibitory capacities for α-glucosidase and α-amylase, respectively (ideal conditions employed in antidiabetic therapy). The study indicates that hydro-alcoholic infusion and maceration were the most appropriate methods to obtain liqueurs with higher values of the aforementioned parameters and safe levels of toxic furanics.
  • Multi-element characterisation of carob, fig and almond liqueurs by MP-AES
    Publication . Rodríguez Solana, Raquel; Dias Carlier, Jorge; Clara Costa, Maria; Romano, Anabela
    Carob pod, fig and almond liqueurs are produced in various Mediterranean regions. This work reports the mineral content of these traditional beverages and evaluates the influence of the raw material. Twenty-five fruit liqueurs from 16 producers were analysed. A simple open-vessel sample mineralization by wet digestion using nitric acid-hydrogen peroxide (1:1) was used before spectrometric analysis. Nine essential elements (Cu, Ca, Mg, Na, K, Fe, Zn, Mn and P) and two non-essentials (Cd and Pb) were quantified by microwave plasma-atomic emission spectrometry (MP-AES). Carob liqueur presented the broader profile of minerals and was the only fruit liqueur containing iron (72.7% of samples), phosphorous, manganese and low levels of lead (in two of the 11 samples). Conversely, almond liqueurs contained the lowest mineral content with only five elements detected. Fruit liqueurs showed variability in mineral content even within the same liqueur, presumably as a consequence of the different manufacturing processes. Despite this variability, application of principal component analysis to essential mineral concentrations (K, Na, Ca, Mg, Mn, Fe and Zn) resulted in satisfactory classification (PC1 and PC2 accounting for 78.5% of the total variance) of the Portuguese liqueurs evaluated.
  • Impact of temperature on Phenolic and Osmolyte contents in In Vitro cultures and micropropagated plants of two mediterranean plant species, Lavandula viridis and Thymus lotocephalus
    Publication . Mansinhos, Inês; Gonçalves, Sandra; Rodríguez Solana, Raquel; Ordóñez-Díaz, José Luis; Moreno-Rojas, José Manuel; Romano, Anabela
    In 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.