Determination of falcarinol in carrot (Daucus carota L.) genotypes using liquid chromatography/mass spectrometry

A new analytical method for the determination of falcarinol [(Z)-heptadeca-1,9-diene-4,6-diyn-3-ol] in carrot root samples has been developed and validated. The method consists of accelerated solvent extraction (ASE) of lyophilised carrot root samples with ethyl acetate and LC–MS analysis of the extracts. Falcarinol was determined by extracting the main ion species generated in the ESI positive mode, m/z 268 [M+H–H₂O+MeCN]⁺, from the full MS chromatogram. Quantitation was performed using a falcarinol calibration curve (correlation coefficient 0.9975) as an external standard and pelargonic acid vanillylamide as an internal standard. The method showed good precision with interday and intraday variation of less than 4% and high recovery (average recovery rate 97.9%). LOD (S/N = 3) and LOQ (S/N = 10) were 2.5 and 7 ng, respectively. Using this method, 27 different carrot genotypes grown and harvested under the same conditions were analyzed, and falcarinol contents ranging from 0.70 to 4.06 mg/100 g fresh weight were determined.     

Dose-effect study on the antioxidant properties of leaves and outer bracts of extracts obtained from Violetto di Toscana artichoke

Artichoke (Cynara scolymus L.) is an edible vegetable largely used in the Mediterranean diet and in folk medicine. The present paper discusses the analysis of the polyphenol content of leaves and outer bracts of Violetto di Toscana artichoke using different extraction procedures with the aim of establishing a correlation between polyphenol subclasses and antioxidant activity measured on human LDL oxidized by copper ions. HPLC/DAD and HPLC/MS analyses revealed that both the matrixes contain identical polyphenol subclasses, with mainly quantitative differences. The antioxidant effect of four artichoke extracts decreases in the following order when the sum of total phenolic compounds was considered: ethanolic extract from leaves (IC₅₀ = 2.92 ± 0.46 μM); ethanolic extract from outer bracts (IC₅₀ = 4.04 ± 0.21 μM); ethyl acetate extract from leaves (IC₅₀ = 4.91 ± 0.11 μM); ethyl acetate extract from outer bracts (IC₅₀ = 10.18 ± 1.6 μM). IC₅₀ were also calculated considering the concentrations of single polyphenol subclasses. In both cases, the potency of antioxidant properties was not related to the amount of total polyphenols or the single subclasses.     

Antioxidant and antimicrobial activity of Cynara cardunculus extracts

The whole, fresh involucral bracts of cardoon, Cynara cardunculus L. (Compositae), were extracted with EtOH and an aqueous suspension of the obtained EtOH extract was partitioned successively with CHCl₃, EtOAc and n-BuOH, leaving a residual water extract. All obtained extracts were evaluated for their antioxidant and antimicrobial properties. The antioxidant potential was evaluated using following in vitro methods: FRAP (ferric reducing antioxidant power) assay, and scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. Antimicrobial activity was estimated using a microdilution technique against food-borne, mycotoxin producers and human pathogenic bacteria and micromycetes. The following bacteria were tested: Salmonella typhimurium, Escherichia coli, Bacillus subtilis, Staphylococcus epidermidis, Staphylococcus aureus, as well as micromycetes: Aspergillus niger, Aspergillus ochraceus, Aspergillus flavus, Penicillium ochrochloron, Penicillium funiculosum, Trichoderma viride, Fusarium tricinctum and Alternaria alternata. Results showed that all extracts possessed concentration-dependent antioxidant activity. In biological assays, C. cardunculus extracts showed antimicrobial activity comparable with standard antibiotics.     

Phlorotannins of the edible brown seaweed Ecklonia cava Kjellman induce sleep via positive allosteric modulation of gamma-aminobutyric acid type A-benzodiazepine receptor: A novel neurological activity of seaweed polyphenols

The primary objective was to investigate whether seaweeds have hypnotic activity. Methanol extracts of 30 seaweeds were screened for their binding activity at the GABA type A-benzodiazepine (GABA_A-BZD) receptor, a well-characterised molecular target for sedative-hypnotics. The most active seaweed was Ecklonia cava Kjellman (ECK). An ethanol extract of ECK (ECK-E) significantly potentiated pentobarbital-induced sleep in mice. In four solvent fractions separated from ECK-E, hypnotic activity was proportional to contents of total phenols and total phlorotannins, known as seaweed polyphenols. Major phlorotannins of the ethyl acetate (EtOAc) fraction with the highest activity were eckol, eckstolonol, dieckol, and triphlorethol-A, and their K_i (binding affinity, μM) values for [³H]-flumazenil binding were 1.070, 1.491, 3.072, and 4.419, respectively. Hypnotic effects of ECK-E and the EtOAc fraction were fully inhibited by flumazenil, a specific GABA_A-BZD receptor antagonist. These results imply that phlorotannins of ECK induce sleep by positive allosteric modulation of the GABA_A-BZD receptor.     

Chemical studies of anthocyanins: A review

Anthocyanins are natural colorants which have raised a growing interest due to their extensive range of colours, innocuous and beneficial health effects. Despite the great potential of application that anthocyanins represent for food, pharmaceutical and cosmetic industries, their use has been limited because of their relative instability and low extraction percentages. Currently, most investigations on anthocyanins are focused on solving these problems, as well as their purification and identification.     

Suppressive effect of ethyl acetate extract of Teucrium polium on cellular oxidative damages and apoptosis induced by 2-deoxy-d-ribose: Role of de novo synthesis of glutathione

The effect of ethyl acetate (EtOAc) extract of Teucrium polium on 2-deoxy-d-ribose (dRib)-induced oxidative stress and apoptosis in erythroleukemic K562 cells was investigated. K562 cells exposed to dRib (50 mM) exhibited abnormal properties such as overproduction of reactive oxygen species, lipid peroxidation, glutathione (GSH) depletion and biochemical features of apoptosis. Treatment with EtOAc extract (25 and 50 μg/ml) reduced oxidative stress and apoptosis among the dRib-treated K562 cells. To disclose the mode of action, the effect of the extract on the cellular GSH and its synthesis by γ-glutamylcysteine synthetase (γ-GCS), a key rate-limiting enzyme of the GSH synthetic pathway, were evaluated. The results demonstrated that the antioxidant property of EtOAc extract mainly results from increasing the level of cellular GSH by inducing the activity of γ-GCS. It can be concluded that the EtOAc extract of T. polium prevents dRib-induced oxidative stress and apoptosis mostly through antioxidative mechanisms.     

UPLC-PDA–MS evaluation of bioactive compounds from leaves of Ilex paraguariensis with different growth conditions,treatments and ageing

Teas from Maté have been widely consumed for centuries in Brazil, Argentina, Paraguay, and Uruguay. We now determine how age, growth conditions and post-harvesting processes of leaves from Ilex paraguariensis affect the concentration of bioactive compounds and their antioxidant capacity. Phenolics, xanthines, and carbohydrates were identified and quantified by electrospray ionisation mass spectrometry (ESI-MS) and ultra performance liquid chromatography (UPLC), which dramatically reduces the time for each analysis (<3 min). On average, sun-exposed (monoculture) leaf extracts exhibited higher levels of bioactive compounds as compared to shaded (forest grown) ones. PCA (principal component analysis) analysis of all the samples indicated that those obtained after blanching and drying contained more phenolics and a smaller concentration of xanthine than those in natura. The oxidised leaves had lower concentrations of phenolics, and consequently a decline in antioxidant activity. No differences were found based on the leaf age.     

塔拉粉中单宁含量的测定

为了充分利用引种资源塔拉,使用络皮粉-紫外分光光度法对其中有效成分单宁的含量分布进行了测定,结果表明在乙酸乙酯中有88.59%的单宁含量。     

Isolation of caffeic acid from Perilla frutescens and its role in enhancing γ-glutamylcysteine synthetase activity and glutathione level

Perilla frutescens is an annual herbaceous plant native to Asia, where its leaves are used in Asian gourmet food. Our previous study showed that the inhibition of γ-glutamylcysteine synthetase (γ-GCS) activity was remarkably recovered by pretreatment with perilla leaf extract (PLE). The objective was to fractionise PLE, and to identify the active component that is responsible for the enhancement of γ-GCS activity and glutathione (GSH) concentration. Among the five fractions from PLE, PLE-III of the ethyl acetate fraction showed the highest γ-GCS activity in a HepG2 cell experiment, and was further chromatographed. The purified compound, which enhanced γ-GCS activity, was finally identified as caffeic acid. We first report the enhancement of γ-GCS activity and GSH level in HepG2 cells by caffeic acid obtained from PLE. Our results suggest that caffeic acid may be a key factor in the chemopreventive potential of perilla leaf components by increasing de novo synthesis of GSH.