Valorisation, Green Extraction Development, and Metabolomic Analysis of Wild Artichoke By-Product Using Pressurised Liquid Extraction UPLC-HRMS and Multivariate Data Analysis.

Valorisation of food by-products has recently attracted considerable attention due to the opportunities to improve the economic and environmental sustainability of the food production chain. Large quantities of non-edible parts of the artichoke plant (Cynara cardunculus L.) comprising leaves, stems, roots, bracts, and seeds are discarded annually during industrial processing. These by-products contain many phytochemicals such as dietary fibres, phenolic acids, and flavonoids, whereby the most challenging issue concerns about the recovery of high-added value components from these by-products. The aim of this work is to develop a novel valorisation strategy for the sustainable utilisation of artichoke leaves' waste, combining green pressurised-liquid extraction (PLE), spectrophotometric assays and UPLC-HRMS phytochemical characterization, to obtain bioactive-rich extract with high antioxidant capacity. Multivariate analysis of the major selected metabolites was used to compare different solvent extraction used in PLE.

Seasonal variation in bioactive properties and phenolic composition of cardoon (Cynara cardunculus var. altilis) bracts.

Cardoon (Cynara cardunculus L.) bracts were collected at different maturation stages to investigate seasonal changes in the phenolic compounds profile and in vitro bioactivities. Among the 12 phenolic compounds tentatively identified, 3,5-O-dicaffeoylquinic acid (21.83 mg/g extract) and apigenin-7-O-glucuronide (10.6 mg/g extract) were the most abundant. Immature bracts (C1: principal growth stage (PGS) 5) had the highest phenolic compounds content, and anti-inflammatory (IC50 = 72 µg/mL) and cytotoxic (GI50 of 30-79 µg/mL) activities. Moreover, extract C1 inhibited efficiently the formation of thiobarbituric acid reactive substances (TBARS; IC50 = 26.8 µg/mL), while extract C8 (PGS 8/9) was more effective against oxidative haemolysis (IC50 38 and 75 µg/mL). The highest antibacterial and antifungal activities were attributed to samples C1 and C6 (PGS 7/8) and samples C2 (PGS 5/6) and C4 (PGS 6/7), respectively. Overall, the obtained results suggest the seasonal changes of polyphenolic composition and bioactivity of cardoon bracts of variable maturity.

Effect of Shading on the Sesquiterpene Lactone Content and Phytotoxicity of Cultivated Cardoon Leaf Extracts.

The work described here follows on from a previous study focused on the influence of the genotype and harvest time on the sesquiterpene lactone (STL) profile of Cynara cardunculus L. leaf extracts. The aim of this study was to investigate the effect that 60% plant shading in cultivated cardoon (C. cardunculus var. altilis) leaf extracts harvested in winter and spring had on the composition of STLs and the phytotoxicity. The phytotoxicity of leaf extracts was evaluated by assessing wheat coleoptile elongation along with seed germination and the root and shoot length of the weeds Amaranthus retroflexus L. and Portulaca oleracea L. Shading increased the production of STLs in spring, and this effect correlated positively with the phytotoxic activity. The induction of shading can therefore be used to modulate STL concentrations and their phytotoxic potential in cultivated cardoon leaves for industrial applications.

Phenolic Compounds in Poorly Represented Mediterranean Plants in Istria: Health Impacts and Food Authentication.

Phenolic compounds are well-known bioactive compounds in plants that can have a protective role against cancers, cardiovascular diseases and many other diseases. To promote local food development, a comprehensive overview of the phenolic compounds' composition and their impact on human health from typical Mediterranean plants such as Punica granatum L., Ziziphus jujuba Mill., Arbutus unedo L., Celtis australis L., Ficus carica L., Cynara cardunculus var. Scolymus L. is provided. Moreover, the potential use of these data for authenticity determination is discussed. Some of the plants' phenolic compounds and their impact to human health are very well determined, while for others, the data are scarce. However, in all cases, more data should be available about the content, profile and health impacts due to a high variation of phenolic compounds depending on genetic and environmental factors. Quantifying variation in phenolic compounds in plants relative to genetic and environmental factors could be a useful tool in food authentication control. More comprehensive studies should be conducted to better understand the importance of phenolic compounds on human health and their variation in certain plants.

Influence of Genotype and Harvest Time on the Cynara cardunculus L. Sesquiterpene Lactone Profile.

The excessive and inappropriate application of herbicides has caused environmental pollution. The use of allelochemicals as bioherbicides could provide a solution to this problem. The allelopathic activity of Cynara cardunculus L. has been studied previously, and sesquiterpene lactones (STLs) were identified as the most relevant allelochemicals. The goal of the study reported here was to investigate the effect of six genotypes and three harvest times on the qualitative and quantitative composition of STLs in C. cardunculus leaves through a new ultra-high-performance liquid chromatography-tandem mass spectrometry analysis method and, thus, the effect on phytotoxicity. Overall, wild cardoon contained the highest levels of STLs of the three botanical varieties studied. Nevertheless, climatic conditions had a marked influence on the presence of STLs among the six genotypes, which was higher in the April harvest. Cynaropicrin was the most abundant STL detected. A close relationship was found between the STL profiles and the allelopathic activity, expressed as inhibition of wheat coleoptile elongation. The data provide a new and important contribution to our understanding of C. cardunculus allelopathy.

Pre-treatment of corn stover, Cynara cardunculus L. stems and wheat straw by ethanol-water and diluted sulfuric acid: Comparison under different energy input conditions.

Ethanol-water (EW) and diluted sulfuric acid (DSA) pre-treatment have been studied for lignocellulosic biomass (corn stover, Cynara cardunculus L. stems and wheat straw). Both pre-treatments have been compared taken into account: solids recovery, glucans recovery, xylans removed, delignification and glucose yield. In all cases, the amount of energy involved has been taken as a criterion for sustainability. In general terms, EW is more efficient to remove lignin and DSA more appropriate to hydrolysate xylans. The combined effect of delignification and xylans removal is responsible for the improvement in the enzymatic cellulose hydrolysis. Under conditions of moderate-low energy inputs, EW pre-treatment yields better results than DSA with glucose yields in the range of 50-60% for EW pre-treated corn stover and cardoon stems; while wheat straw pulps reach up to 80%. So, multiple raw materials biorefinery needs a previous study to fit the type and conditions of the pre-treatment to each feedstock.

Haplotype analysis of the germacrene A synthase gene and association with cynaropicrin content and biological activities in Cynara cardunculus.

Cynara cardunculus: L. represents a natural source of terpenic compounds, with the predominant molecule being cynaropicrin. Cynaropicrin is gaining interest since it has been correlated to anti-hyperlipidaemia, antispasmodic and cytotoxicity activity against leukocyte cancer cells. The objective of this work was to screen a collection of C. cardunculus, from different origins, for new allelic variants in germacrene A synthase (GAS) gene involved in the cynaropicrin biosynthesis and correlate them with improved cynaropicrin content and biological activities. Using high-resolution melting, nine haplotypes were identified. The putative impact of the identified allelic variants in GAS protein was evaluated by bioinformatic tools and polymorphisms that putatively lead to protein conformational changes were described. Additionally, cynaropicrin and main pentacyclic triterpenes contents, and antithrombin, antimicrobial and antiproliferative activities were also determined in C. cardunculus leaf lipophilic-derived extracts. In this work we identified allelic variants with putative impact on GAS protein, which are significantly associated with cynaropicrin content and antiproliferative activity. The results obtained suggest that the identified polymorphisms should be explored as putative genetic markers correlated with biological properties in Cynara cardunculus.

Effects of saline stress on mineral composition, phenolic acids and flavonoids in leaves of artichoke and cardoon genotypes grown in floating system.

BACKGROUND: Cynara cardunculus is a species native to the Mediterranean basin. It includes globe artichoke and cultivated cardoon as well as their progenitor wild cardoon. The species is a source of biophenols, and its leaf extracts have been widely used in herbal medicine as hepatoprotectors and choleretics since ancient times. The aim of this study was to determine the effect of increasing the level of salinity in the nutrient solution (1 or 30 mmol L⁻¹ NaCl) on biomass production, mineral composition, radical-scavenging activity, caffeoylquinic acids and flavonoids in three artichoke ('Romolo', 'Violetto di Provenza' and 'Violetto di Romagna') and three cultivated cardoon ('Bianco Avorio', 'Bianco Gigante Inerme' and 'Gigante di Romagna') cultivars grown in a floating system. RESULTS: Increased salinity in the nutrient solution decreased the leaf dry biomass and leaf number of artichoke and cultivated cardoon cultivars. Salinity reduced macro- and microelement accumulation in leaves (e.g. N, K, Ca, Mg, Fe, Mn and B) but improved their antioxidant activity, total polyphenols, chlorogenic acid, cynarin and luteolin. The cultivated cardoons, especially 'Bianco Avorio' and 'Gigante di Romagna', showed higher biomass and leaf number than those observed in artichoke genotypes. 'Violetto di Provenza' exhibited the highest content of chlorogenic acid, closely followed by 'Violetto di Romagna', whereas for cynarin content the highest values were recorded in 'Violetto di Provenza', 'Bianco Avorio' and 'Gigante di Romagna'. The highest content of luteolin was recorded in 'Gigante di Romagna' and 'Bianco Avorio', while the highest content of apigenin was observed in 'Gigante di Romagna'. CONCLUSION: The results showed that the floating system could be considered an effective tool to improve quality aspects through proper management of the salt concentration in the nutrient solution. They also suggest that specific cultivars should be selected to obtain the desired profile of bioactive compounds.

Responses of Cynara cardunculus L. to single and combined cadmium and nickel treatment conditions.

A greenhouse pot experiment was carried out to study the responses of Cynara cardunculus L. (cardoon) to cadmium and nickel. Three groups of 12 pots each were planted with cardoon plants and spiked with single and combined cadmium and nickel aqueous solutions. The bioavailable metal concentrations, measured in soil, ranged widely and were up to 246.7 mg kg⁻¹ for Cd and 61.1 mg kg⁻¹ for Ni. Under Cd treatment, cardoon growth remained unaffected, while increased Ni soil concentrations inhibited plant growth and were lethal to the highly treated plants. In the combined Cd and Ni treatments, an antagonistic effect was observed between the two metals. Cadmium and nickel concentrations in cardoon tissues rose with increasing metal concentrations in the soil. Cadmium and nickel contents in shoots reached 169.3 and 342.3 mg kg⁻¹ in the single treatments while, under the combined Cd and Ni treatments, they were up to 235.0 and 440.7 mg kg⁻¹, respectively. Generally, mean contents of both metals in the shoots were higher than in the roots and the translocation factor was greater than 1. A possible enhancing effect of nickel on cadmium uptake was observed. Cardoon showed characteristics of a Cd accumulator.

Phenolic composition of Cynara cardunculus L. organs, and their biological activities.

Polyphenols are bioactive molecules exhibiting a lot of scientific attention due to their multiple biological activities. This study compared phenolic contents and antioxidant activity in Cynara cardunculus L. organs and focus on leaf phenolic compounds identification by RP-HPLC and their antibacterial activity. The analyzed organs exhibited different total polyphenol contents (7-14.8 mg GAE g(-1) DW). Leaf and seed phenolic contents were similar and two times higher than those in flowers. The same tendency was observed for the amount of flavonoids and tannins. However, seed extracts displayed the highest DPPH. scavenging ability with the lowest IC50 value (23 microg ml(-1)), followed by leaves and flowers (over 50 microg ml(-1)). In contrast, leaves showed the highest capacity to quench superoxide (IC50: 1 microg ml(-1)) as compared to seeds (6 microg ml(-1)). In addition, cardoon leaves were efficient to inhibit growth of pathogenic bacteria mainly against Staphylococcus aureus and Escherichia coli. The identification of phenolic compounds from leaves revealed that syringic and trans-cinnamic acids were the major molecules.