Antioxidant activity and protective effect of the outer scales hydroalcoholic extract of Allium cepa L. var. Tropea on toxicity damage induced by Cadmium in Caco-2 cells.

The characterization of bioactive metabolites and the protective effects against Cadmium (Cd) of the hydroalcoholic extract from Allium cepa var. Tropea (Tropea red onion) in human Caco-2 colon adenocarcinoma cells and in vitro antioxidant effects were investigated. Tropea red onion extract showed high levels of bioactive compounds and a strong activity as radical scavenger and inhibitor of lipid peroxidation. The most abundant specialized metabolites were quercetin derivatives. In addition, the extract inhibited NO release in a dose-dependent manner. Exposure of Cd treated human Caco-2 cells to the Tropea red onion extract resulted in a higher cytoprotection with a significant falloff of cells damage produced by Cd. The effect of Tropea red onion evaluated with the MTT assay and LDH test, was time-dependent and more evident after 24h of treatment. Caco-2 cells treated with the Tropea red onion extract and CdCl2 at 24h showed a significant cytoprotection at concentrations equal to 50 µg/mL + 25 µM and 100 µg/mL + 25 µM while at 72h at a concentration of 25 µg/mL + 25 µM. We observed with MTT and LDH assays that treatment of cells with Tropea red onion extract at 24h was able to significantly prevent Cd-induced cytotoxicity alone.

Digestive and gastroprotective effects of Achillea erba-rotta subsp. moschata (Wulfen) I.Richardson (syn. A. moschata Wulfen) (Asteraceae): From traditional uses to preclinical studies.

ETHNOPHARMACOLOGICAL RELEVANCE: Achillea erba-rotta subsp. moschata (Wulfen) I.Richardson (syn. A. moschata Wulfen) (Asteraceae) is an alpine endemic plant whose aerial parts are harvested by the locals mainly for the digestive properties. Despite its widespread use, few studies have been conducted to date to verify its bioactivity. AIM OF THE STUDY: The purpose of the work was to meet the tradition confirming with experimental data the popular belief that the consumption of this species offers beneficial effects to the gastrointestinal system. MATERIALS AND METHODS: Using Soxhlet apparatus, the dried aerial parts of A. erba-rotta subsp. moschata were successively extracted with petroleum ether (PET), dichloromethane (DCM) and methanol (MeOH). The essential oil (EO) was obtained by hydrodistillation using a Clevenger apparatus while infusion (AE) was prepared following the traditional local recipe. Their chemical characterization was performed by various techniques including SPME-GC/MS, GC/MS and HPLC/MS-MS. An in vitro biological screening was carried out. The influence of AE on lipid digestion was monitored by titration of free fatty acids (FFA) during pancreatic lipase activity with the pH-stat method. For all extracts and EO, the anti-Helicobacter pylori activity was assessed by the broth microdilution method, the influence on cell viability was evaluated against NCI-N87, OE21 and Caco-2 cell lines and a preliminary toxicity evaluation was done using Brine Shrimp lethality (BSL) assay. The anti-inflammatory potential was evidenced by interleukin IL-1- induced IL8 expression on Caco-2 cells. RESULTS: AE increased by 15% the FFA releasing compared to the pancreatic lipase alone. PET, DCM and MeOH extracts as well as AE and EO were considered active against the growth of both antimicrobial susceptible and resistant strains of H. pylori with MIC values starting from 16 µg/mL. PET and DCM (IC50 = 89 µg/mL and 96 µg/mL, respectively, against Caco-2 cell line) extracts showed the high effect on cell viability while the EO reduced in 50% of cell viability at 1.48 µL/mL (NCI-N87 cells), 1.42 µL/mL (OE21 cells), and 3.44 µL/mL (Caco-2 cells) corroborating the BSL results. In different degrees, all extracts and EO inhibited the IL-1ß-stimulated IL-8 production in Caco-2 cells. CONCLUSIONS: The obtained data are encouraging and provide a scientific basis for the traditional use of A. erba-rotta subsp. moschata as a digestive agent although they need to be further corroborated by studies involving the investigation of both the in vivo activities and the role of the compounds detected in the extracts.

Polyphenol content and bioactivity of Achillea moschata from the Italian and Swiss Alps.

Achillea moschata Wulfen, which grows in the Alps, is extensively used by local people for its medicinal properties. Two studied samples were collected, at the flowering stage, in Val Mustair (Switzerland) and Valchiavenna (Italy), respectively. The aerial parts were defatted with petroleum ether (PET) and successively extracted with dichloromethane (DCM) and methanol (MeOH). High-performance liquid chromatography and electrospray ionization-tandem mass spectrometry analyses of the methanolic extracts evidenced that flavonoids were the predominant compounds compared to phenolic acids in both samples (89.5 vs. 33.0 µg/mg DW in A. moschata Valchiavenna and 82.5 vs. 40.0 µg/mg DW in A. moschata Val Mustair). Among flavonoid derivatives, luteolin and apigenin were the predominant aglycones, free and glycosilated. The A. moschata Valchiavenna extract was characterized by apigenin as the main compound (60.4 µg/mg DW), while A. moschata Val Mustair was characterized by its derivative apigenin 7-O-glucoside (44.7 µg/mg DW). The antioxidant activity of all the obtained extracts was tested by the DPPH (2,2-diphenyl-picryl hydrazyl) and ABTS (2,21-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) methods, which showed their increasing scavenger capacity in relation to extract polarity (PET extract < DCM extract < MeOH extract). The extracts were also investigated against three Gram-positive (Bacillus cereus, Enterococcus faecalis and Staphylococcus aureus) and three Gram-negative (Escherichia coli, Proteus mirabilis and Pseudomonas aeruginosa) bacterial species using the disc diffusion assay. DMC and PET were the most active extracts (inhibition diameter: ≥12 mm).

Quality Assessment of Commercial Spagyric Tinctures of Harpagophytum procumbens and Their Antioxidant Properties.

Preparations from the dried tubers of Harpagophytum procumbens (Burch.) DC ex Meisn, commonly known as devil's claw, are mainly used in modern medicine to relieve joint pain and inflammation in patients suffering from rheumatic and arthritic disorders. This paper describes for the first time the chemical profile of a commercial spagyric tincture (named 019) prepared from the roots of the plant. For comparison purposes, a commercial not-spagyric devil's claw tincture (NST) was also analyzed. Chemical investigation of the content of specialized metabolites in the three samples indicated that harpagoside was the main compound, followed by the two isomers acteoside and isoacteoside. Compositional consistence over time was obtained by the chemical fingerprinting of another spagyric tincture (named 014) from the same producer that was already expired according to the recommendation on the label of the product. The two spagyric preparations did not show significant compositional differences as revealed by HPLC and MS analyses, except for a decrease in harpagide content in the expired 014 tincture. Moreover, their antioxidant capacities as assessed by 2,2'-di-phenyl-1-picrylhydrazyl (DPPH) and 2.2'-azin-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) methods resulted in very similar IC50 values. The expired 014 tincture showed instead a lower IC50 value compared to the 019 and NST tinctures with the ferric reducing antioxidant potential (FRAP) assay, indicating a higher ferric-reducing antioxidant ability. Overall, these results indicated that the two preparations could generally maintain good stability and biological activity at least for the four years from the production to the expiration date.

Phytochemical and Biological Profile of Moricandia arvensis (L.) DC.: An Inhibitor of Pancreatic Lipase.

Pancreatic lipase, a key enzyme for lipid absorption, is one of the most important targets for the treatment of obesity, while natural compounds have recently attracted much interest as potential inhibitors of this enzyme. Here, in an attempt to find new effective agents, the methanolic extract from Moricandia arvensis (L.) DC. and its sub-extracts were investigated for their potential inhibitory activity. The ability to inhibit pancreatic lipase was verified through the in vitro evaluation of the prevention of p-nitrophenyl caprylate hydrolysis. The antioxidant activity was also verified by means of DPPH and ß-carotene bleaching tests. Compositional profiling revealed that flavonoid glycosides were the main specialized metabolites present in the methanolic extract from the aerial parts of the plant with kaempferol and quercetin representing the two O-glycosylated aglycones. Kaempferol-3-O-ß-(2″-O-glucosyl)-rutinoside and kaempferol-3-O-a-arabinosyl-7-O-rhamnoside were the most abundant flavonols. The crude methanolic extract and the dichloromethane and ethyl acetate sub-extracts showed a strong lipase inhibitory activity, with IC50 values of 2.06 ± 0.02, 1.52 ± 0.02 and 1.31 ± 0.02 mg/mL, respectively. The best capacity to scavenge DPPH radical was detected for the ethyl acetate sub-extract (IC50 = 171.9 ± 1.0 µg/mL), which was also effective in protecting linoleic acid from peroxidation (IC50 = 35.69 ± 2.30 µg/mL). Obtained results support the hypothesis that M. arvensis can be a source of bioactive phytochemicals for the pharmacological inhibition of dietary lipids absorption.

Nematicidal potential of Taraxacum officinale.

This study was aimed to investigate the activity of the Asteraceae species Taraxacum officinale against the root-knot nematode Meloidogyne incognita. Leaf and root extracts of T. officinale were tested in vitro at a range of 62.5-1000 and 250-1000 µg mL-1 concentrations on nematode juveniles and eggs, respectively, whereas treatments with 10-40 g kg-1 soil rates of dry leaf and root T. officinale biomass were applied to soil infested by M. incognita in greenhouse experiments on potted tomato. Peak 36 and 50% juvenile mortality and 14.8 and 23.8% egg hatchability reduction were recorded at the maximum concentration of leaf and root extracts, respectively. Soil treatments with T. officinale leaf and root material strongly suppressed nematode multiplication and gall formation on tomato roots and significantly increased plant growth. Chicoric acid and 3-O- and 3,5-di-O-caffeoylquinic acid were found to be the main components of leaf and root extract, respectively, and proved, as the total hydroalcoholic extracts from T. officinale leaf and root material, for an antioxidant activity. Data from this study indicate the suitability of plant materials from T. officinale for a potential formulation of nematicidal products to include in sustainable nematode management strategies.

Chemical Profile, Antioxidant and Antibacterial Activities of Achillea moschata Wulfen, an Endemic Species from the Alps.

Aerial parts of Achillea moschata Wulfen (Asteraceae) growing wild in the Italian Rhaetian Alps were investigated to describe, for the first time, their phenolic content, as well as to characterize the essential oil. Inspection of the metabolic profile combining HPLC-DAD and ESI-MS/MS data showed that the methanol extract contained glycosylated flavonoids with luteolin and apigenin as the main aglycones. Among them, the major compound was 7-O-glucosyl apigenin. Caffeoyl derivates were other phenolics identified. The essential oil obtained by steam distillation and investigated by GC/FID and GC/MS showed camphor, 1,8-cineole, and bornylacetate as the main constituents. The antioxidant capacity of three different extracts with increasing polarity and of the essential oil was evaluated by employing ABTS·+ and DPPH· radical scavenging assays. The methanolic extract was the only significantly effective sample against both synthetic radicals. All samples were also tested against Gram-positive (Bacillus cereus, Enterococcus faecalis, Staphylococcus aureus) and Gram-negative (Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa) bacterial species using the disk diffusion assay. The non-polar extracts (dichloromethane and petroleum ether) and the essential oil possessed a broad spectrum of antimicrobial activity expressed according to inhibition zone diameter (8-24 mm).

Compositional Analysis of Lavandula pinnata Essential Oils.

The genus Lavandula includes about thirty species plus a number of intraspecific taxa and hybrids, which are distributed in the Mediterranean area. The traditional use of lavender both as perfume or medicinal plant is known since antiquity. Nowadays several species are extensively cultivated for the extraction of their essential oils (EOs) which are used in manufactured products like cosmetics and perfumes or in phytotherapy. Lavandula pinnata L. f. (syn L. pinnata Lundmark) is a rare species native to the Canary Islands used in folk medicine as relaxant and also a valuable remedy against bites. To the best of our knowledge, EOs from L. pinnata have been very little studied. The present paper reports on the quali- and quantitative compositional profile of the EOs distilled (by a Spring type apparatus) from the aerial parts (flowers and leaves) of this species cultivated in soilless conditions. Chemical analyses by means of GC and GC-MS techniques have indicated that oxygenated monoterpenes are the main constituents of both the flowers (68.30%) and the leaves (83.65%). Carvacrol is the main compound which characterizes the EOs of this species. In addition, discrete amounts of spathulenol (12.22%) and caryophyllene oxide (14.62%) have been detected in flowers EOs, while leaves EOs contained small amounts of carvacrol methyl ether (2.52%).

Phytochemical analysis of Passiflora loefgrenii Vitta, a rich source of luteolin-derived flavonoids with antioxidant properties.

OBJECTIVES: The paper describes the flavonoid composition of the aerial parts (young leaves, YL; adult leaves, AL; stems, ST) of Passiflora loefgrenii Vitta, a rare species native to Brazil, where it is traditionally used as food. Antioxidant potential has also been evaluated. To the best of our knowledge, no phytochemical and biological study on this species has been reported previously. METHODS: Compositional data have been acquired combining HPLC-diode array detector (DAD) and Electrospary ionization-tandem mass spectrometry (ESI-MS/MS) analyses. Antioxidant activity has been evaluated by the 2,2'-di-phenyl-1-picrylhydrazyl method. KEY FINDINGS: Glycosylated flavones, with luteolin as the main aglycone, can be regarded as biomarkers for this drug. Qualitative composition of the extracts from YL, AL and ST was similar. The bulk of the constituents was made up by 8-C-ß-glucosyl luteolin (orientin), 7-O-α-rhamnosyl-6-C-ß-glucosyl luteolin and 6-C-α-rhamnosyl luteolin, which totally amounted to 16.57 (73%), 10.77 (74%) and 5.07 (77%) µg/mg in YL, AL and ST, respectively. P. loefgrenii showed a good antioxidant activity (IC50 of 350 µg/ml), higher than generally reported for other passifloras. CONCLUSIONS: P. loefgrenii, rich in luteolin glycosides, can be regarded as a good candidate to be explored for therapeutic properties other than the sedative one since it represents a rich source of valuable flavonoids with antioxidant potential.

Phytochemical analysis of a herbal tea from Artemisia annua L.

Strategies to control diffusion of malaria needs to account for the increase of resistance of the parasite to the conventional antimalarial drugs. It has been proposed that a traditional aqueous preparation from Artemisia annua, with a low content of the active compound, artemisinin, may reduce the risk of resistance of the protozoa and be relatively more effective in the treatment of the disease. The solubility properties of the molecule have been the matter of concern about the therapeutic usefulness of herbal teas from A. annua. The present study aimed at analysing the chemical profile of a tea infusion from A. annua. Tea from A. annua was prepared through infusion of the plant aerial parts in water for 1, 24 and 48 h. Content of artemisinin was determined by HPLC-ELSD. Overall chemical characterization of the extracts was carried out by a combination of metabolomic techniques. The artemisinin content varied only slightly in the three different extracts (about 0.12%). A series of mono-caffeoyl- and mono-feruloyl-quinic acids, di-caffeoyl- and di-feruloyl-quinic acids was identified as main components of the tea infusion, together with some flavonoids. Reconstitution of the same extracts in less polar or apolar solvents resulted in a different composition with no phenolics and a much lower concentration of artemisinin.

Glucosinolates profile of "mugnolo", a variety of Brassica oleracea L. native to southern Italy (Salento).

Glucosinolates (GLSs) from a variety of Brassica oleracea, known locally as "mugnolo" and widely distributed in southern Italy, were studied. It was found that "mugnolo" inflorescences are characterized by the presence of the aliphatic GLSs glucoraphanin (1.79 µmol/g), glucoiberin, glucoerucin, and sinigrin, of the aromatic GLSs glucobarberin (0.56 µmol/g) and gluconasturtin, and of the indole GLSs glucobrassicin (3.51 µmol/g), neoglucobrassicin, 4-methoxyglucobrassicin, and 4-hydroxyglucobrassicin. Indole GLSs were predominant, while aliphatic and aromatic GLS were found in lower quantities. The metabolic profile of "mugnolo" leaves was also studied. It was found that 4-hydroxyglucobrassicin (0.13 µmol/g) and glucoraphanin (0.11 µmol/g) are the predominant GLSs. Vegetables of the Brassicaceae family are seldom consumed raw; therefore we also analyzed the GLS profile of "mugnolo" after cooking in water. The results showed variations in the GLSs content with a fall in concentration of 50%.

In vitro synergic efficacy of the combination of Nystatin with the essential oils of Origanum vulgare and Pelargonium graveolens against some Candida species.

In this study we investigated a synergistic effect between the essential oils Origanum vulgare, Pelargonium graveolens and Melaleuca alternifolia and the antifungal compound Nystatin. Nystatin is considered a drug of choice in the treatment of fungal infections, but it can cause some considerable problems through its side effects, such as renal damage. Finding a new product that can reduce the Nystatin dose via combination is very important. Our findings showed an experimental occurrence of a synergistic interaction between two of these essential oils and Nystatin. The essential oil O. vulgare appeared to be the most effective, inhibiting all the Candida species evaluated in this study. Some combinations of Nystatin and P. graveolens essential oil did not have any synergistic interactions for some of the strains considered. Associations of Nystatin with M. alternifolia essential oil had only an additive effect.

Triterpenoid glycosides from leaves of Medicago arborea L.

Eighteen triterpene saponins (1-18) from Medicago arborea leaves have been isolated and their structures elucidated by spectroscopic, spectrometric (1D and 2D NMR, FAB-MS, ESI-MS/MS), and chemical methods. They have been identified as glycosides of medicagenic, zanhic, and 2beta-hydroxyoleanolic acids, soyasapogenol B, bayogenin, and 2beta,3beta-dihydroxyolean-12-en-23-al-28-oic acid. Twelve of them, identified as 3-O-beta-D-glucopyranosyl-28-O-[alpha-L-arabinopyranosyl(1-->3)-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside] zanhic acid (3), 3-O-beta-D-glucopyranosyl-28-O-[beta-D-xylopyranosyl(1-->4)-[alpha-L-arabinopyranosyl-(1-->3)]-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside] zanhic acid (4), 3-O-[alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranosyl(1-->2)-beta-D-glucopyranosyl]-2beta-hydroxyoleanolic acid (5), 3-O-beta-D-glucuronopyranosyl-28-O-[alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside]medicagenic acid (6), 3-O-beta-D-glucuronopyranosyl-28-O-[beta-D-xylopyranosyl(1-->4)-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside]bayogenin (9), 3-O-beta-D-glucuronopyranosyl-28-O-[beta-D-xylopyranosyl(1-->4)-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside]-2beta,3beta-dihydroxyolean-12-en-23-al-28-oic acid (10), 3-O-beta-D-glucuronopyranosyl-28-O-[beta-D-xylopyranosyl(1-->4)-[beta-D-apiofuranosyl(1-->3)]-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside]zanhic acid (12), 3-O-beta-D-glucuronopyranosyl-28-O-[beta-D-xylopyranosyl(1-->4)-[alpha-L-arabinopyranoside(1-->3)]-alpha-L-rhamnopyrano-syl(1-->2)-alpha-L-arabinopyranoside]zanhic acid (13), 3-O-beta-D-glucuronopyranosyl-28-O-[beta-D-xylopyrano-syl(1-->4)-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside]zanhic acid (14), 3-O-[alpha-L-arabinopyranosyl-(1-->2)-beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranosyl]-28-O-[beta-D-xylopyranosyl(1-->4)-[beta-D-apiofurano-syl(1-->3)]-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside]zanhic acid (16), 3-O-[beta-D-glucopyrano-syl(1-->2)-beta-D-glucopyranosyl]-28-O-[beta-D-xylopyranosyl(1-->4)-[alpha-L-arabinopyranosyl(1-->3)]-alpha-L-rhamno-pyranosyl (1-->2)-alpha-L-arabinopyranoside]zanhic acid (17), and 3-O-beta-D-glucuronopyranosyl-28-O-[beta-D-xylopyranosyl(1-->4)-[beta-D-apiofuranosyl(1-->3)]-alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyrano-side]medicagenic acid (18), are reported as new natural compounds. The presence of the aldehydic group on the sapogenin moiety of saponin 10 is discussed in the framework of a possible elucidation of the biosynthesis of these metabolites.