Invasive Alien Species as a Potential Source of Phytopharmaceuticals: Phenolic Composition and Antimicrobial and Cytotoxic Activity of Robinia pseudoacacia L. Leaf and Flower Extracts.

Black locust (Robinia pseudoacacia L.), an invasive tree in Europe, commonly known for its negative impact on biodiversity, is a rich source of phenolic compounds recognized in traditional medicine. Since the metabolite profile depends on the environment and climate, this study aimed to provide the first LC-MS phytochemical screening of the black locust from the Istria region (Croatia). The compounds were extracted from leaves and flowers with 70% ethanol and 80% methanol. Total phenolics (TP) and flavonoids (TF), as well as antioxidant capacity (AC) measured by ABTS (17.49-146.41 mg TE/g DW), DPPH (24.67-118.49 mg TE/g DW), and FRAP (7.38-77.53 mg TE/g DW) assays, were higher in leaf than in flower extracts. Higher TP and total non-flavonoid (TNF) values were displayed in ethanolic than in methanolic extracts. In total, 64 compounds were identified, of which flavonols (20) and hydroxycinnamic acid derivatives (15) were the most represented. Flavanols such as catechin dominated in leaf extracts, followed by flavonols, with kaempferol glucuronyl rhamnosyl hexosides as the main compound, respectively. Flower extracts had the highest share of flavones, followed by ellagitannins, with luteolin dirhamnosyl hexosides and vescalagin, respectively, being predominant. The extracts had good quorum sensing, biofilm formation prevention, and eradicating capacity. The results provided new insights into the phytochemical properties of R. pseudoacacia as the first step toward its potential pharmaceutical use.

LC-DAD-MS Phenolic Characterisation of Six Invasive Plant Species in Croatia and Determination of Their Antimicrobial and Cytotoxic Activity.

Invasive plants' phytochemicals are important for their invasiveness, enabling them to spread in new environments. However, these chemicals could offer many pharmaceutical compounds or active ingredients for herbal preparations. This study provides the first LC-MS phytochemical screening of six invasive alien plant species (IAPS) in the Istria region (Croatia): Ailanthus altissima, Ambrosia artemisiifolia, Conyza canadensis, Dittrichia viscosa, Erigeron annuus, and Xanthium strumarium. The study aims to identify and quantify the phenolic content of their leaf extracts and assess their antimicrobial and cytotoxic potential. A total of 32 species-specific compounds were recorded. Neochlorogenic, chlorogenic, and 5-p-coumaroylquinic acids, quercetin-3-glucoside, and kaempferol hexoside were detected in all the tested IAPS. Hydroxycinnamic acid derivatives were the main components in all the tested IAPS, except in E. annuus, where flavanones dominated with a share of 70%. X. strumarium extract had the best activity against the tested bacteria, with an average MIC value of 0.11 mg/mL, while A. altissima and X. strumarium extracts had the best activity against the tested fungi, with an average MIC value of 0.21 mg/mL in both cases. All the plant extracts studied, except X. strumarium, were less cytotoxic than the positive control. The results provided additional information on the phytochemical properties of IAPS and their potential for use as antimicrobial agents.

Changes in beneficial bioactive compounds in eight traditional herbal liqueurs during a one-month maceration process.

BACKGROUND: Herbs are rich in various beneficial bioactive compounds and they can be used for many different purposes. One of the most common is maceration of herbs in alcohol. Different substances respond differently to extraction in prepared solutions. It is very important to enhance the highest proportion of beneficial ingredients during the maceration process in a herbal liqueur. A comparative analysis of numerous different phenolic compounds from eight of the most frequently used herbs for making aperitifs in Europe was performed. RESULTS: In the comparison among all studied herbs, the highest phenolic content was found with Artemisia absinthium (wormwood) (14.61 mg g-1 ) and Mentha piperita (peppermint) (13.89 mg g-1 ), while the lowest content was found with Centaurium erythraea (common centaury) (3.96 mg g-1 ). Salvianolic acid isomers and caffeic acid derivatives were the greatest contributors to the total phenolic content in lemon balm, wormwood, peppermint, fennel and sage. These compounds contain more hydroxyl groups and they were better extracted at the beginning of the maceration process. Caraway and common centaury contain more flavonols (quercetin and kaempferol derivatives), with higher chemical stability and fewer hydroxyl groups in their structure. CONCLUSION: The compositions of eight herbal liqueurs were highly related to the presence of different herbal ingredients and their solubility in the extract over time. Most flavonol derivatives were extracted over a longer time and the two liqueurs (common centaury and caraway) therefore had the highest phenolic contents after 3 weeks of maceration. © 2019 Society of Chemical Industry.

Which Plant Part of Purple Coneflower (Echinacea purpurea (L.) Moench) Should be Used for Tea and Which for Tincture?

Medicinal plants are widely used for the relief of disease symptoms or as dietary supplements. In recent decades, purple coneflower has become extremely well known. An infusion or tincture of purple coneflower can be prepared by anyone simply, inexpensively, and ecologically safely. Three plant parts of purple coneflower were used in the study: extracts from roots, flowers, and leaves were obtained using three different solvents (100% and 40% ethanol and water). High-performance liquid chromatography-mass spectrophotometer identified and quantified 23 individual phenolics. Pure (100%) ethanol gave the lowest yield of all the investigated phenolic compounds in all herb parts. Chicoric and caftaric acids were the major phenolic compounds in coneflower. Caftaric acid, with health promoting properties, was extracted best in a water solution from purple coneflower leaves (2673.31 mg/100 g dry weight [DW]) and chicoric acid, also with a beneficial effect on human health, yielded the highest levels in 40% ethanol solution from flowers (1571.79 mg/100 g DW) and roots (1396.27 mg/100 g DW).

Traditional rose liqueur - A pink delight rich in phenolics.

Chemical composition and colorimetric parameters of alcoholic liqueurs prepared from rose petals were evaluated by comparing the potential of three cultivars ('Amadeus', 'Colossal Meidiland' and 'Rosanna') and three traditional methods of preparation (fresh/air-dried petals extracted in 50% ethanol or aqueous sucrose syrup). Extraction was performed at room temperature for 2 weeks. High performance liquid chromatography/mass spectrometry was used to confirm the presence of 6 anthocyanins, 4 flavanols, 4 phenolic acids, 2 hydrolysable tannins and 31 flavonols in petal liqueurs. The highest concentrations of anthocyanins were determined in extracts from 'Amadeus' petals, followed by 'Colossal Meidiland' and finally, 'Rosanna'. The best extraction yields and optimal colour characteristics were achieved by ethanolic extraction of dry petals followed by fresh petal extraction in ethanol and, finally, extraction in sucrose syrup. Air-dehydration of 'Amadeus' petals prior to extraction in 50% ethanol yielded rose liqueur with the best all round characteristics.

Changes in phenolic profiles of red-colored pellicle walnut and hazelnut kernel during ripening.

In studies of secondary metabolites in nuts, many constituents in the kernel remain unidentified due to a high content of phenolic compounds in the pellicle. In the present study, we focused on the investigation of the phenolic and dicarboxylic acid profiles of walnut and hazelnut pellicle-less kernels. High-performance liquid chromatography with diode array and mass spectrometric detection (HPLC-DAD-MSn) was used to carry out the determination of individual phenolics and dicarboxylic acids in brown and red-pellicle walnut and hazelnut. Results show that hexahydroxydiphenic acid (HHDP) di-galloyl hexose isomer, vanillic acid hexoside, quinic acid derivative and catechin are the main constituents of the phenolic profile of walnut, while galloylquinic derivative, caffeoyl hexoside and catechin are the main constituents of the hazelnut kernel. Even though both walnut and hazelnut kernels have a considerably lower content of phenolic compounds and dicarboxylic acids in comparison to the pellicles, when calculated as a percentage of the total mass, the kernel makes a significant contribution to the total phenolic content of the whole nut.

Fruit Seeds of the Rosaceae Family: A Waste, New Life, or a Danger to Human Health?

In fruit production seeds are mostly regarded as waste, but for plants they represent a beginning of new life. Seeds accumulate toxic or health-beneficial compounds, and the elucidation of their metabolic profile is especially important to people who consume the entire fruit, including the seeds. The present research quantifies the levels of bioactive compounds (phenolics and cyanogenic glycosides (CGG)) in fruit seeds of 35 cultivars belonging to 6 different fruit species. High-performance liquid chromatography and mass spectrophotometry were used to detect and identify the studied compounds. Significant differences in the content of individual bioactive compounds as well as their groups were recorded (p < 0.05). For the first time neoamygdalin and prunasin were detected in a number of fruit cultivars. All fruit seeds, except pears, accumulated from 2- to 46-fold higher levels of CGG than phenolics. On average, seeds contained from 75.46 to 1648.14 µg/g phenolics and from 46.39 to 4374.31 µg/g CGG. The study also clarifies the new lethal dose for cyanogenic glycosides.

Bioactive Components and Antioxidant Capacity of Fruits from Nine Sorbus Genotypes.

In this study, biochemical profile of fruits of 9 Sorbus genotypes was analyzed. The content of total sugars ranged from 69.7 g/kg ('Titan') to 217.5 g/kg (Sorbus torminalis) and total organic acids from 17.7 g/kg ('Businka') to 40.2 g/kg (S. torminalis). The highest content of total anthocyanins had 'Burka' (871 mg/kg FW) and 'Businka' (856 mg/kg FW). Quercetin derivatives represented more than 95% of total flavonols. 'Alaja krupnaja' had 3.5- to 29-fold higher rutin content than other analyzed genotypes. S. torminalis fruits had the greatest diversity of isorhamnetin and kaempferol derivatives. Chlorogenic acid was the major hydroxycinnamic acid and its share was 33% to 73% of total analyzed hydoxycinnamic acid derivatives. The richest in chlorogenic acid were 'Krasavica' and 'Alaja krupnaja' fruits. Cultivar 'Businka' had the highest content of epicatechin (40.7 mg/kg) and neochlorogenic acid (1061 mg/kg). Different procyanidin oligomers were detected among flavanols in Sorbus fruits. The highest content of total flavanols was measured in 'Alaja Krupnaja' fruits. Cultivars 'Krasavica' (84.5 mg/kg) and 'Burka' (85.1 mg/kg) had 1.2- to 6.9-fold higher amount of total carotenoids. 'Businka' was highlighted as the richest in total tannin and phenolic contents (3768 mg GAE/kg) and consequently, it had the highest antioxidant activity (57.6 mM TE/kg FW). Being abundant in polyphenolics, some extracts of Sorbus genotypes, for example, 'Businka,' 'Burka,' and 'Alaja krupnaja' could serve as valuable resource of bioactive compounds to food and pharmaceutical industries.

The higher the better? Differences in phenolics and cyanogenic glycosides in Sambucus nigra leaves, flowers and berries from different altitudes.

BACKGROUND: Elderberry (Sambucus nigra L.) possesses high antioxidant activity and has been used to treat numerous medicinal disorders. In addition to their antioxidant properties, elderberry parts accumulate toxic cyanogenic glycosides (CGG). It has been proven that altitude influences the biosynthesis of many secondary metabolites. In the present study we investigated the change of phenolics and CGG in elder leaves, flowers, and berries induced by different altitudes and locations. RESULTS: The data indicate that the accumulation of CGG and phenolics is affected by the altitude of the growing site. An increase of anthocyanin content was recorded in elder berries collected at higher elevations in both locations. Fruit collected at the foothills of location 2 contained 3343 µg g-1 anthocyanins as opposed to fruit from the hilltop, which contained 7729 µg g-1 . Elder berries contained the lowest levels of harmful CGG compared to other analysed plant parts. However, more cyanogenic glycosides were always present in plant parts collected at the hilltop. Accordingly, berries accumulated 0.11 µg g-1 CGG at the foothills and 0.59 µg g-1 CGG at the hilltop. CONCLUSION: Elder berries and flowers collected at the foothill were characterised by the lowest levels of both beneficial (phenolics) and harmful compounds (CGG) and are suitable for moderate consumption. © 2016 Society of Chemical Industry.

Wild Prunus Fruit Species as a Rich Source of Bioactive Compounds.

Sugars, organic acids, carotenoids, tocopherols, chlorophylls, and phenolic compounds were quantified in fruit of 4 wild growing Prunus species (wild cherry, bird cherry, blackthorn, and mahaleb cherry) using HPLC-DAD-MSn. In wild Prunus, the major sugars were glucose and fructose, whereas malic and citric acids dominated among organic acids. The most abundant classes of phenolic compounds in the analyzed fruit species were anthocyanins, flavonols, derivatives of cinnamic acids, and flavanols. Two major groups of anthocyanins measured in Prunus fruits were cyanidin-3-rutinoside and cyanidin-3-glucoside. Flavonols were represented by 19 derivatives of quercetin, 10 derivatives of kaempferol, and 2 derivatives of isorhamnetin. The highest total flavonol content was measured in mahaleb cherry and bird cherry, followed by blackthorn and wild cherry fruit. Total phenolic content varied from 2373 (wild cherry) to 11053 mg GAE per kg (bird cherry) and ferric reducing antioxidant power antioxidant activity from 7.26 to 31.54 mM trolox equivalents per kg fruits.

White versus blue: Does the wild 'albino' bilberry (Vaccinium myrtillus L.) differ in fruit quality compared to the blue one?

Wild albino and blue bilberry fruit were analyzed to compare different fruit characteristics linked to the composition of primary and secondary metabolites. Compounds were identified using HPLC-MS and standard quality parameters were determined. Albino berries were significantly smaller, accumulated less water and were characterized by 23% higher dry weight. Soluble solids content and pH value were higher in albino bilberry and their surface was lighter and characterized by a yellow hue. Both bilberry forms accumulated identical individual sugars and organic acids; however, the albino form had 33% higher content of total sugars and 9% higher content of total organic acids compared to the blue type. Fifteen anthocyanins were identified in both forms, but in albino bilberries, individual anthocyanins were only detected in traces. Blue bilberry contained 1.6-fold higher levels of flavanols, 2.1-fold higher levels of flavonols, 2.5-fold higher levels of hydroxycinnamic acid derivatives and consequently, 4.6-fold higher total phenolic content.

Comparison of major taste compounds and antioxidative properties of fruits and flowers of different Sambucus species and interspecific hybrids.

Differences in the content of sugars, organic acids, total phenolics and antioxidative activity have been evaluated among three different elderberry species (Sambucus nigra, Sambucus cerulea, Sambucus javanica) and seven interspecific hybrids. The highest content of sugars has been determined in the fruits of JA×CER hybrid and the lowest in fruits of (JA×NI)×cv. Black Beauty hybrid. S. nigra berries contained highest levels of total organic acids. S. nigra and (JA×NI)×CER flower extracts were characterized by 1.3- to 2.8-fold higher content of total sugars compared to other species/hybrids analyzed. Total phenolic content (TPC) in berries ranged from 3687 to 6831 mg GAE per kg FW. The highest TPC has been determined in S. nigra fruits and flowers. The ABTS scavenging activity differed significantly among species and hybrids and ranged from 3.2 to 39.59 mM trolox/kgF W in fruits and 44.87-118.26 mM trolox/kg DW in flowers.

Fresh from the Ornamental Garden: Hips of Selected Rose Cultivars Rich in Phytonutrients.

Morphological parameters (size, weight, color), the content of sugars, organic acids, lycopene, ß-carotene, and phenolics were determined in hips of Rosa canina (RCA), Rosa sweginzowii (RSW), Rosa rugosa (RUG), and selected ornamental Rosa cultivars Fru Dagmar Hastrup (FDH), Repandia (REP), Veilchenblau (RVB), Aloha (RAL), Bonica (BON), and Golden Gate (RGG). Although traditionally used RCA hips contained the highest amount of cyanidin-3-glucoside (83 µg/g DW) and were the reddest (h° = 17.5), they did not stand out in other analyzed parameters. RGG climber had the biggest hips (8.86 g), which also contained highest sugar levels (50.9 g/100 g DW). RAL stood out as the cultivar rich in organic acids (33.9 g/100 g DW), mainly because of high quinic acid content (17.6 g/100g DW). FDH and RSW hips were characterized by particularly high ascorbic acid levels (4325 mg/100 g DW and 4711 mg/100 g DW). Other ornamental cultivars contained low amounts of ascorbic acid compared to the analyzed species. The phenolic profile was species/cultivars-specific. The greatest diversity of phenolic compounds was detected in RUG and FDH hips (55 and 54 different tentatively identified compounds with HPLC/MS). Flavanols represented the main phenolic class in most of the investigated species/cultivars and RGG hips contained the highest amount of catechin and proanthocyandin derivatives (15855 µg/g DW). Altogether RAL hips contained the highest quantity of phenolics (44746 µg/g DW) mainly due to high levels of hydrolysable tannins compared to other species/cultivars. Although small, hips of BON and REP were most abundant regarding ß-carotene and lycopene content, respectively.

Fruit Phenolic Composition of Different Elderberry Species and Hybrids.

UNLABELLED: The aim of this study was to investigate a detailed composition and content of phenolic compounds in fruits of 4 elderberry species (Sambucus nigra, S. cerulea, S. ebulus, and S racemosa) and 8 interspecific hybrids. Hydroxycinnamic acids (HCAs) represented the major share of phenolics in analyzed elderberries; caffeoylquinic and p-coumaroylquinic acids were most abundant. Flavanols (catechin, epicatechin, and different procyanidins) were the second major phenolic group detected in range from 2% to 30% of total analyzed phenolics. From the group of flavonols, 13 different quercetin glycosides, 7 kaempferol glycosides, and 8 isorhamnetin glycosides have been quantified. Rutin was the major flavonol in all studied genotypes. S. ebulus was characterized by the highest level of total HCAs, catechin, epicatechin, and most flavonols. Some elderberry hybrids, for example JA × RAC, CER × NI, and JA × (JA × NI), are perspective for further studies because they have high content of phenolic compounds. The results of research could contribute to breed cultivars, which may prove interesting for food-processing industries. PRACTICAL APPLICATION: Different levels of phenolic compounds have been measured in fruit of analyzed elderberry species and interspecific hybrids. Natural evaluation of elderberry genetic resources, the identification of phenolic compounds, and assessment of their properties are of great interest to breeders and directly valid for pharmaceutical and food industry.

Comparison of phenolic profiles and antioxidant properties of European Fagopyrum esculentum cultivars.

The purpose of this study was to investigate composition and content of phenolic compounds in seeds of common buckwheat (Fagopyrum esculentum Moench) cultivars from Western, Central and Southeastern Europe grown in the Balkan area, and to compare them with cultivars from the Balkan. Mostly detected hydroxycinnamic acids in seeds of the investigated cultivars were caffeic and chlorogenic acid derivatives. More than ten different flavanols were detected in the investigated seeds, based on which all tested buckwheat cultivars were divided into two groups: those with high propelargonidins (epiafzelechin-epicatechin) and those with high procyanidins contents. 'Novosadska' had the highest level of phenolic acids, proanthocyanidins, flavones and most of the flavonols. However, 'Bosna 1' and 'Bosna 2' were highlighted with the greatest rutin content (up to 46 times higher than in other cultivars). All buckwheat cultivars had quite high antioxidant capacity (more than 80% of neutralized radicals), yet, 'Novosadska', 'Godijevo', 'Spacinska 1' and 'Bamby' excelled.

Traditional elderflower beverages: a rich source of phenolic compounds with high antioxidant activity.

Traditional elderflower beverages are very popular due to their pleasing taste and pro-health properties. Phenolics were quantified in three elderflower extracts (methanolic extraction and two water extracts prepared as fresh drinks according to local recipes) with HPLC-MS. Chromatographic patterns of methanolic and water extracts were quite similar; however, content levels of individual compounds differed between the two extraction solvents. Two feruloylquinic acids and two p-coumaroyl-caffeoylquinic acids and different flavonol glycosides have been identified in elderflowers for the first time. Hydroxycinnamic acids and flavonol glycosides were the major phenolic constituents in elderflowers. Methanolic extract contained higher levels of all phenolic groups compared to water-extracted samples. The yield of elderflower extracts depended on extract solution and time of extraction. Elderflower beverages were characterized by high total phenolic content (91.09 to 746.63 mg GAE/L). The results confirmed that elderflower beverages (sabesa, syrup) should be recommended as a good source of phenolics in human diet.

Changes in fruit quality parameters of four Ribes species during ripening.

There are no defined maturity parameters for berry fruit which often results in harvesting and marketing of inedible, immature or over ripe berries. In the present study the changes in quality characteristics of different white-, red-, and black-currant cultivars as well as gooseberry and jostaberry fruit were investigated at three maturity stages. Colour parameters, berry juice pH level and the content of sugars, organic acids and phenolic compounds have been evaluated. A total of 65 different phenolic compounds were detected by HPLC-PDA/MS. The results indicate that, during the last 2-4 weeks of fruit ripening, significant changes occur in colour parameters, primary and secondary metabolites and radical scavenging activity. The results suggest that the average content of specific phenolic groups and sugars/acids ratio can be used as a simple and reliable maturity index for optimisation of harvest time.

High concentrations of anthocyanins in genuine cherry-juice of old local Austrian Prunus avium varieties.

Antioxidant activity and polyphenols were quantified in vapour-extracted juice of nine Austrian, partially endemic varieties of sweet cherry (Prunus avium): cv. 'Spätbraune von Purbach', cv. 'Early Rivers', cv. 'Joiser Einsiedekirsche', cv. 'Große Schwarze Knorpelkirsche' and four unidentified local varieties. Additionally the effect of storage was evaluated for six of the varieties. A variety showing the highest antioxidant capacity (9.64 µmol Trolox equivalents per mL), total polyphenols (2747 mg/L) and total cyanidins (1085 mg/L) was suitable for mechanical harvest and its juice did not show any losses of antioxidant capacity and total anthocyanin concentration during storage. The juice of cv. 'Große Schwarze Knorpelkirsche' had also high concentrations of total anthocyanins (873 mg/L), but showed substantial losses through storage. The local Austrian sweet cherry varieties from the Pannonian climate zone are particularly suitable for the production of processed products like cherry juice with high content of anthocyanins and polyphenols.

Novel roles for the polyphenol oxidase enzyme in secondary metabolism and the regulation of cell death in walnut.

The enzyme polyphenol oxidase (PPO) catalyzes the oxidation of phenolic compounds into highly reactive quinones. Polymerization of PPO-derived quinones causes the postharvest browning of cut or bruised fruit, but the native physiological functions of PPOs in undamaged, intact plant cells are not well understood. Walnut (Juglans regia) produces a rich array of phenolic compounds and possesses a single PPO enzyme, rendering it an ideal model to study PPO. We generated a series of PPO-silenced transgenic walnut lines that display less than 5% of wild-type PPO activity. Strikingly, the PPO-silenced plants developed spontaneous necrotic lesions on their leaves in the absence of pathogen challenge (i.e. a lesion mimic phenotype). To gain a clearer perspective on the potential functions of PPO and its possible connection to cell death, we compared the leaf transcriptomes and metabolomes of wild-type and PPO-silenced plants. Silencing of PPO caused major alterations in the metabolism of phenolic compounds and their derivatives (e.g. coumaric acid and catechin) and in the expression of phenylpropanoid pathway genes. Several observed metabolic changes point to a direct role for PPO in the metabolism of tyrosine and in the biosynthesis of the hydroxycoumarin esculetin in vivo. In addition, PPO-silenced plants displayed massive (9-fold) increases in the tyrosine-derived metabolite tyramine, whose exogenous application elicits cell death in walnut and several other plant species. Overall, these results suggest that PPO plays a novel and fundamental role in secondary metabolism and acts as an indirect regulator of cell death in walnut.

Comparison of phenolic composition of healthy apple tissues and tissues affected by bitter pit.

Bitter pit is an important Ca(2+) deficiency disorder of apple fruit (Malus domestica Borkh.), with symptoms, necrotic spots, developing during storage. The objective of this study was to determine phenolic compounds and their contents in bitter pit in comparison to healthy skin and pulp using HPLC-MS(2). The experiment was carried out on three cultivars 'Jonagored', 'Golden Delicious' and 'Pinova'. All 15 determined phenolic compounds in pulp tissues specifically affected by bitter pit were higher than those in healthy pulp. Chlorogenic acid and catechin were to 5 times higher in those affected pulp tissues. Higher content was also determined for hydroxycinnamic acids and flavanols in the peel above the bitter pit; in contrast, flavonols and anthocyanins were higher in healthy peel. Anthocyanins in healthy peel of cultivar 'Jonagored' were 10 times higher from the content in peel above the bitter pit.