Cupressus sempervirens L., Juniperus communis L. and Cistus ladanifer L. are Mediterranean arboreal and shrub species that possess essential oils (EO) in their leaves and branches. This study aimed at characterizing the EOs obtained by steam distillation from the three species collected in different locations from Spain (Almazán, Andévalo, Barriomartín, Cerezal, Ermitas and Huéscar). For this purpose, volatiles composition was determined by GC-MS, and different bioactivities were evaluated. The highest content in terpenes was observed in C. sempervirens (Huéscar origin) followed by J. communis (Almazán origin), corresponding to 92% and 91.9% of total compounds, respectively. With exception of C. ladanifer from Cerezal that presented viridiflorol as the most abundant compound, all the three species presented in common the α-pinene as the major compound. The EOs from C. ladanifer showed high antibacterial potential, presenting MIC values from 0.3 to 1.25 mg/mL. Concerning other bioactivities, C. ladanifer EO revealed an oxidation inhibition of 83%, while J. communis showed cytotoxicity in the MCF-7 cell line, and C. sempervirens and C. ladanifer EOs exhibited the highest potential on NCI-H460 cell lines. Nevertheless, some EOs revealed toxicity against non-tumoral cells but generally presented a GI50 value higher than that of the tumor cell lines.

The vibrant colours of many plants are due to secondary metabolites, such as nitrogen-containing compounds, where betacyanins are included. These compounds can be found in plants such as Amaranthus caudatus L. that, due to their high nutritional benefits, have been overproduced, which leads to the accumulation of large amounts of bio-residues. Among these bio-residues, the flowers which have a very intense pink colour and present no economic value or subsequent destination can be exploited as sources of natural colouring agents (betacyanins). This work aimed at characterising the flower’s extract in terms of bioactive molecules such as tocopherols, organic acids, but essentially in terms of betacyanins, in order to obtain a natural colouring agent. For the extraction, ultrasound-assisted extraction (UAE) ideal conditions were obtained using the Response Surface Methodology (RSM), allowing the attainment of an enriched extract of betacyanins in high yields and purity. The obtained extracts were analysed for their bioactive potential, namely antioxidant, antimicrobial and cytotoxic properties. From the obtained results, three isoforms of tocopherols were detected, β-tocopherol (0.884 ± 0.003 mg/100 g dry weight (dw)) being the most abundant one. Regarding the organic acids, oxalic (2.48 ± 0.05 mg/100 g dw), shikimic (0.170 ± 0.003 mg/100 g dw) and traces of fumaric acid were found. Four betacyanins were identified and quantified, namely: amaranthine (171 ± 1 mg/g extract), isoamaranthine (38 ± 1 mg/g extract), betanin (1.6 ± 0.1 mg/g), and isobetanin (1.3 ± 0.1 mg/g extract). The obtained extract also presented antioxidant activity with inhibition concentration (IC50 values) of 29.0 ± 0.4 μg/mL and 114 ± 4 μg/mL for Δt of 60 min and 120 min, respectively in the oxidative haemolysis inhibition assay (OxHLIA) assay. The obtained extract also presented an interesting antibacterial activity with minimum inhibitory concentrations ranging from 5 to 20 mg/mL against pathogenic bacteria and revealed no toxicity for normal cells.

Essential oils (EOs) obtained from Juniperus communis L. are frequently used in the production of bioproducts. However, there are no studies regarding industrial crops’ production, allowing for better control of the quality and production of juniper EOs. To select the plant material for developing future crops of this species in northern Spain, four locations where this shrub species grows in the wild were selected and samples of both genera were collected. The EOs were obtained by steam distillation, and their chemical composition and bioactivity were evaluated. The results showed that the yield of EOs from male and female samples were within the usual reported ranges, varying between 0.24 and 0.58% (dry basis, d.b.). However, limonene content in three locations varied between 15 and 25%, which is between 100% and 200% higher than the values usually reported for other European countries. The antibacterial activity was determined by broth microdilution and showed that gram-positive bacteria were more susceptible to the tested EOs since, in general, lower MIC values were obtained compared to gram-negatives. The EOs from location 1 (L1F) and 2 (L2M) inhibited the growth of six out of the eight clinical strains tested. Samples from location 1 were particularly effective, exhibiting MBC against two gram-negative (E. coli and P. mirabilis) and one gram-positive bacteria (E. faecalis). Moreover, the majority of the EOs tested showed anti-inflammatory activity. Cytotoxic effect has been demonstrated in tumor cell lines, with the best results observed against gastric carcinoma (AGS) cells (GI50 between 7 to 77 µg/mL). Although generally presenting higher GI50, most samples also inhibited the growth of non-tumoral cells, particularly hepatocytes (PLP2 cells). Therefore, its use for their anti-proliferative activity should consider specific conditions to avoid damaging normal cells. Finally, the results and conclusions obtained led to the selection of the female shrubs from location 1 (L1F) as the plant material to be propagated in order to produce plants for a future juniper crop.

Consumer demand for natural and healthier products has led to an increasing interest in the bioactive and therapeutic properties of plant extracts. In this study, we evaluated the phenolic compounds profile, bioactivities, and toxicities of plant extracts from eight European flora species, including Calendula officinalis L., Calluna vulgaris (L.) Hull, Hippophae rhamnoides L., Juglans regia L., Mentha cervina L., Rubus idaeus L., Sambucus nigra L., and Vitis vinifera L. The aim was to identify potential preservatives of natural origin. Phenolic compounds were identified by HPLC-DAD-ESI-MS. Caffeic acid derivatives, ellagitannins, flavonols, and flavones were the major phenolic compounds identified. The total phenolic content varied from 16.0 ± 0.2 (V. vinifera) to 123 ± 2 mg/g (H. rhamnoides) of dry extract. All extracts showed antioxidant potential and exhibited activity against some of the microorganisms tested. S. nigra showed the highest activity in the inhibition of oxidative hemolysis (OxHLIA) assay and H. rhamnoides, notably, had the lowest IC50 values in TBARS and DPPH assays, as well as the lowest minimum inhibitory concentration (MIC) values. Regarding in vitro cytotoxicity, in tumor and non-tumor cell lines, although some extracts revealed toxicity against normal cells, it was found that the samples C. vulgaris, V. vinifera and R. idaeus might be used against tumor cells since the active concentration is much lower than the one causing toxicity. In vivo acute toxicity tests using Artemia franciscana suggest low toxicity for most extracts, with LC50 > 400 mg/L. These results showed the potential of the studied extracts as natural preservatives, given their richness in compounds with bioactive properties, highlight their potential value to the production chain.

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