Proanthocyanidin profile and antioxidant capacity of Brazilian Vitis vinifera red wines

The free flavan-3-ol and proanthocyanidin (PA) profile and the antioxidant capacity of wines Vitis vinifera L., 2006 and 2007 vintages, from the São Joaquim region, at southern Brazil, are reported here for the first time. Catechin and epicatechin were the two main monomers in the wine samples, followed by gallocatechin and epigallocatechin; and the PA B1 was the main dimer. The terminal units of the PAs were constituted mainly by catechin units, with the co-presence of epicatechin, gallocatechin, epigallocatechin and traces of epicatechin gallate. The epicatechin and epigallocatechin units were the main constituents of the extension units of PAs with the co-presence of catechin and epicatechin gallate. The values for the mean degree of polymerisation ranged from 4.9 to 9.8. The wine samples demonstrated effective scavenging activity against DPPH and ABTS radicals and against lipid peroxidation in vitro. A positive correlation existed between flavan-3-ol content and antioxidant capacity in vitro.     

Inhibition of the angiotensin-converting enzyme by grape seed and skin proanthocyanidins extracted from Vitis vinífera L. cv. País

The influence of two extracts of grape skin and seeds from Vitis vinífera L. cv. País (Chilean black grapes), rich in proanthocyanidins (PAs), was evaluated on the inhibition of the angiotensin I-converting enzyme (ACE), and the inhibition was related to the type and number of subunits of the polymeric PAs chain. Size exclusion chromatography was used to purify the extract and its characterization was made by acid catalysis depolymerization followed by HPLC. ACE activity was measured by quantitative HPLC, measuring the hyppuric acid (HA) produced from the hydrolysis of hippuryl-l-histidyl-l-leucine (HHL) by ACE. Structural compositions differed significantly between both extracts: the skin extracts do not exhibit epicatechin (EC) and epicatechin gallate (ECG), and the seed extracts did not present epigallocatechin (EGC). Skin extracts have a higher mean degree of polymerization (mDP) than seed extracts and a higher inhibition power (IC₅₀ = 0.14 ± 0.03 μM and IC₅₀ = 0.480 ± 0.03 μmol/L), respectively. The catechin (IC₅₀ = 1495 ± 90 μmol/L) and epicatechin (IC₅₀ = 1772 ± 121 μmol/L) monomers exerted lower inhibition than the either grape extract. The structural differences between grape skin and seed PAs could influence the ACE inhibition capacity. The larger inhibitory power of skin extract was associated to greater OH availability, higher mDP and the presence of EGC.     

Study on chemical composition,antioxidant and anti-inflammatory activities of hot water extract from Piceamariana bark and its proanthocyanidin-rich fractions

Hot water extract (HWE) from Picea mariana (black spruce) bark was evaluated for its antioxidant and anti-inflammatory activities. HWE was fractionated into an oligomeric proanthocyanidin-rich fraction (OPF) and a polymeric proanthocyanidin-rich (PPF) fraction, and their bioactivities were also evaluated. OPF exhibited the highest antioxidant and anti-inflammatory activities. The compositions of HWE, OPF and PPF were also evaluated. OPF was richest in total polyphenol but the lowest in total proanthocyanidin (PA) content. Thiolysis coupled with HPLC-DAD has shown that the PAs of the black spruce bark were procyanidin type. The mean degrees of polymerisation (DPm) of HWE, OPF and PPF were, respectively, 6.0, 3.3 and 7.6.     

Binding of polyphenols to plant cell wall analogues – Part 2: Phenolic acids

Bacterial cellulose and cellulose–pectin composites were used as well-defined model plant cell wall (PCW) systems to study the interaction between phenolic acids (PA) derived from purple carrot juice concentrate (PCJC) and PCW components. Significant PA depletion from solution occurred, with pure cellulose initially (30 s–1 h) absorbing more than cellulose–pectin composites in the first hour (ca 20% cf 10–15%), but with all composites absorbing similar levels (ca 30%) after several days. Individual PAs bound to different relative extents with caffeic acid > chlorogenic acid > ferulic acid. Extrapolation of data for these model systems to carrot puree suggests that nutritionally-significant amounts of PAs could bind to cell walls, potentially restricting bioavailability in the small intestine and, as a consequence, delivering PAs to the large intestine for fermentation and metabolism by gut bacteria.     

Jakobs-Kreuzkraut (<Emphasis Type="Italic">Senecio jacobaea</Emphasis>): eine Ursache für Pyrrolizidin-Alkaloide im Sommerhonig?

Pyrrolizidine alkaloids (PAs) are secondary plant compounds, which may be carried into honey by honey bees (Apis mellifera) and thus enter the human food chain. Depending on the quantity consumed, PAs are considered harmful to human health. At present, however, there is no statutory limit for PAs in foodstuffs. Because of several hints that larger occurrences of PA-containing tansy ragwort (Senecio jacobaea) may be responsible for PAs in summer honeys, honey samples from regions with and without tansy ragwort from all across Schleswig-Holstein were tested for their PA content (28 individual PAs) by LC-MS/MS analysis in 2014. By including honey samples from surroundings of flower plots established on arable land, it was tested if the carry-over of PAs into honey can be reduced by “distraction feed”. Samples from regions with tansy ragwort showed significantly higher PA contents than those from regions without tansy ragwort. Flower plots had no significant influence on the PA content. PAs were detected in 53.5 % of the 86 honey samples investigated. The mean PA value of PA-positive honeys was much higher than the median value since the sample comprised only few high and many low values. 28.6 % of the detected PAs were not from Senecio species, but from the Borage family (Boraginaceae) or Hemp Agrimony (Eupatorium cannabinum). In 2.3–15.1 % of the samples the limit value recommended by the BfR (Analytik und Toxizität von Pyrrolizidinalkaloiden sowie eine Einschätzung des gesundheitlichen Risikos durch deren Vorkommen in Honig. Stellungnahme Nr. 038/2011 des BfR vom 11. August 2011, ergänzt am 21. Januar 2013. Bundesinstitut für Risikobewertung [Hrsg.], Berlin: 37 S, 2013a) for different eating habits was exceeded. Because of the potential risk to human health, beekeepers and users of land with larger occurrences of tansy ragwort should take joint measures to keep the PA load in summer honeys as low as possible.     

Preparation of pickering emulsion stabilised by Zein/Grape seed proanthocyanidins binary composite

This paper introduced a kind of binary composite particle constructed by zein particles and grape seed proanthocyanidins (PAs). The physicochemical performance of the zein/PAs complex colloidal particles (ZPAPs) was systematically evaluated. Fourier transform infrared spectroscopy (FTIR) showed that the interactions between zein and PAs were mainly hydrogen bonds and hydrophobic interactions. The three-phase contact angle (θo/w) of zein particles was around 120.7 ± 0.7°, and it was decreased to 100.8 ± 0.6° after the addition of PAs, indicating that ZPAPs were more suitable for stabilising Pickering emulsion. ZPAPs stabilised peanut oil Pickering emulsion (ZPAPEs) at 0.5–0.7 (v/v) oil fractions had no obvious phase separation during a 30-day storage, indicating its good storage stability. Rheological results showed that ZPAPEs had excellent plasticity and viscoelasticity. Confocal laser scanning microscopy (CLSM) showed that particles had stably anchored on the surface of oil droplets which could effectively prevent from Ostwald ripening. The incorporation of PAs to zein endowed zein with stronger affinity, thus enhancing the stability of Pickering emulsion. This research constructed a new novel food-grade emulsifier to prepare Pickering emulsion with potential applications, and further broaden the bioavailability of PAs.     

Chemical fingerprinting identifies Echium vulgare,Eupatorium cannabinum and Senecio spp. as plant species mainly responsible for pyrrolizidine alkaloids in bee-collected pollen

Various studies have shown that bee-collected pollen sold as nutritional supplements may contain toxic pyrrolizidine alkaloids (PAs) and, thus, pose a potential health risk for consumers. The level of contamination may vary according to its geographical and botanical origin. Here, the PA content of pollen produced in Switzerland was studied and 32 commercially available bee-collected pollen supplements produced between 2010 and 2014 were analysed. In addition, at what time period bees collect PA-containing pollen was investigated. Hence, this study looked into the occurrence of PAs in pollen samples collected daily during two-to-three consecutive seasons. Furthermore, the PA spectrum in pollen was compared to the spectrum found in flower heads of PA-plants to unambiguously identify plants responsible for PA contamination of pollen. The PA concentration of commercial and daily collected pollen was determined by target analysis using an HPLC-MS/MS system, allowing the detection of 18 different PAs and PA N-oxides found in the genera Echium, Eupatorium and Senecio, while the comparison of the PA spectrum in pollen and flower heads was performed by LC-HR-MS, allowing the detection of all PA types in a sample, including saturated, non-carcinogenic PAs. Of the commercially available pollen, 31% contained PAs with a mean concentration of 319 ng/g, mainly Echium- and Eupatorium-type PAs, while the PA concentrations were below the limit of quantitation (LOQ) in 69% of the pollen samples. Bees collected pollen containing Echium-type PAs mainly in June and July, while they gathered pollen containing Eupatorium-type PAs from mid-July to August. Senecio-type PAs appeared from June to September. Comparison of the PA array in pollen and plants identified E. vulgare and E. cannabinum as the main plants responsible for PA contamination of Swiss bee-collected pollen, and to a lesser extent also identified plants belonging to the genus Senecio.     

Pyrrolizidine alkaloids in floral honeys of tropical Ghana: a health risk assessment

ABSTRACT

There is a vast amount of information about the nutritional and medicinal properties of honey as a result of its numerous benefits. However, honeys have been found to be contaminated with hepatotoxic and carcinogenic pyrrolizidine alkaloids (PAs) on account of bees foraging on PA-containing plants. This study deals with the analysis of PAs in tropical honeys emanating from different agro-ecological zones of Ghana in order to assess its potential health risk. PAs of 48 honey samples were analysed using high-performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS). The results show that a total of 85% of the honeys from various agro-ecological zones were PA positive including all honeys from supermarkets. The highest concentration of PAs was 2639 μg kg^?1, while the average PA concentration of the samples was 283 μg kg^?1. The study also found Chromolaena odorata pollens in majority of the honeys, thus indicating the plant as major source of PA contamination of honeys in the tropical regions.     

Effects of heat and β-lactoglobulin on distribution of fluorescently labeled tissue- and urokinase-type plasminogen activators in a model milk system

A fluorescent labeling method was developed to determine the effects of heat and β-lactoglobulin (BLG) on the concentration distribution of added fluorescently labeled tissue-type (tPA-647) and urokinase-type (uPA-546) plasminogen activators (PAs) in a model milk system. Prior to heating, the majority of the added PAs (87% of tPA-647, 72% of uPA-546) became associated with casein micelles and could be dissociated by acidification or NaCl. Addition of 0.5% BLG to the unheated system had no significant effect on the distribution of PAs. Heat-induced binding of uPA-546 to micelles was shown by an increase of uPA-546 from 72% to 88% or 95% in the micelle fraction of systems without or with BLG, respectively. Although heat alone had little effect on the total amount of tPA-647 associated with caseins (88%), addition of BLG to a heated system shifted 32% of the tPA-647 to a non-sedimentable casein fraction. After heating, casein–PA interactions became more complex.     

Quantitative analysis of phenolic compounds in Chinese hawthorn (Crataegus spp.) fruits by high performance liquid chromatography–electrospray ionisation mass spectrometry

Eleven major phenolic compounds (hyperoside, isoquercitrin, chlorogenic acid, ideain, epicatechin, two procyanidin (PA) dimers, three PA trimers and a PA dimer-hexoside) were quantified in the fruits of 22 cultivars/origins of three species of the Chinese hawthorn (Crataegus spp.) by HPLC–ESI-MS-SIR. Hyperoside (0.1–0.8 mg/g dry mass [DM]), isoquercitrin (0.1–0.3 mg/g DM), chlorogenic acid (0.2–1.6 mg/g DM), epicatechin (0.9–11.7 mg/g DM), PA B2 (0.7–12.4 mg/g DM), PA dimer II (0.1–1.5 mg/g DM), PA trimer I (0.1–2.7 mg/g DM), PA trimer II (0.7–6.9 mg/g DM), PA trimer III (0.01–1.2 mg/g DM) and a PA dimer-hexoside (trace–1.1 mg/g DM) were detected in all the samples. Ideain (0.0–0.7 mg/g DM) was found in all the samples except Crataegus scabrifolia. Significant correlations between the contents of individual PA aglycons were observed (r > 0.9, P < 0.01). A strong correlation between flavonols was also shown (r = 0.71, P < 0.01). Fruits of Crataegus pinnatifida var. major had higher contents of PAs but lower contents of flavonols compared with Crataegus brettschneideri. The fruits of C. scabrifolia contained the highest level of PA dimer-hexoside, which was present in trace amounts in the fruits of C. pinnatifida.     

Research progress regarding the effect and mechanism of dietary phenolic acids for improving nonalcoholic fatty liver disease via gut microbiota

Phenolic acids (PAs), a class of small bioactive molecules widely distributed in food and mainly found as secondary plant metabolites, present significant advantages such as antioxidant activity and other health benefits. The global epidemic of nonalcoholic fatty liver disease (NAFLD) is becoming a serious public health problem. Existing studies showed that gut microbiota (GM) dysbiosis is highly associated with the occurrence and development of NAFLD. In recent years, progress has been made in the study of the relationship among PA compounds, GM, and NAFLD. PAs can regulate the composition and functions of the GM to promote human health, while GM can increase the dietary sources of PAs and improve its bioavailability. This paper discussed PAs, GM, and their interrelationship while introducing several representative dietary PA sources and examining the absorption and metabolism of PAs mediated by GM. It also summarizes the effect and mechanisms of PAs in improving and regulating NAFLD via GM and their metabolites. This helps to better evaluate the potential preventive effect of PAs on NAFLD via the regulation of GM and expands the utilization of PAs and PA-rich food resources.     

Pyrrolizidine and tropane alkaloids in teas and the herbal teas peppermint,rooibos and chamomile in the Israeli market

Dehydro pyrrolizidine alkaloids (dehydro PAs) are carcinogenic phytotoxins prevalent in the Boraginaceae, Asteraceae and Fabaceae families. Dehydro PAs enter the food and feed chain by co-harvesting of crops intended for human and animal consumption as well as by carry-over into animal-based products such as milk, eggs and honey. Recently the occurrence of dehydro PAs in teas and herbal teas has gained increasing attention from the EU, due to the high levels of dehydro PAs found in commercially available teas and herbal teas in Germany and Switzerland. Furthermore, several tropane alkaloids (TAs, e.g. scopolamine and hyoscyamine) intoxications due to the consumption of contaminated herbal teas were reported in the literature. The aim of the present study was to determine the dehydro PAs and TAs levels in 70 pre-packed teabags of herbal and non-herbal tea types sold in supermarkets in Israel. Chamomile, peppermint and rooibos teas contained high dehydro PAs levels in almost all samples analysed. Lower amounts were detected in black and green teas, while no dehydro PAs were found in fennel and melissa herbal teas. Total dehydro PAs concentrations in chamomile, peppermint and rooibos teas ranged from 20 to 1729 μg/kg. Except for black tea containing only mono-ester retrorsine-type dehydro PAs, all other teas and herbal teas showed mixed patterns of dehydro PA ester types, indicating a contamination by various weed species during harvesting and/or production. The TA levels per teabag were below the recommended acute reference dose; however, the positive findings of TAs in all peppermint tea samples warrant a more extensive survey. The partially high levels of dehydro PAs found in teas and herbal teas present an urgent warning letter to the regulatory authorities to perform routine quality control analysis and implement maximum residual levels for dehydro PAs.     

Detection of pyrrolizidine alkaloids in commercial honey using liquid chromatography–ion trap mass spectrometry

Pyrrolizidine alkaloids (PAs) are known secondary plant metabolites which can cause hepatotoxicity in both humans and livestock. PAs can be consumed through the use of plants for food, medicinal purposes and as contaminants of agricultural crops and food. PA contaminated grain has posed the largest health risk, although any PA contamination in our food chain should be recognised as a potential health threat. For this purpose, retail honeys were tested by LC–MS/MS. The method allows for specific identification of toxic retronecine and otonecine-type PAs by comparison to reference compounds via a spectral library. In total, 50 honey samples were matched to the reference spectra within a set of tolerance parameters. Accurate data analysis and quick detection of positive samples was possible. Positive samples contained an average PA concentration of 1260 μg kg^?1 of honey. Good linear calibrations were obtained (R² > 0.991). LOD and LOQ ranged from 0.0134 to 0.0305 and 0.0446 to 0.1018 μg mL^?1, respectively.     

Transport of biodegradable polymeric particles loaded with grape seed extract across Caco‐2 cell monolayers

Proanthocyanidins (PAs) extracted from grapes have several bioactivities, which have attracted attention for pharmacological uses. However, PA instability in the digestive tract and low intestinal absorption limits their use for oral administration. An alternative to overcome these challenges is encapsulation. In this study, PAs extracted from grape seed were encapsulated in polylactic acid (PLA), polylactic acid with polyethylene glycol (PLA/PEG) and chitosan succinyl lauryl (LSC), and the transport particles and cytotoxicity were evaluated. PLA and PLA/PEG particles had encapsulation close to 80%EE, were spherical in shape, had sizes in the range of 300–400 nm and had a negative zeta potential. LSC particles had an encapsulation of 40%EE, semi‐spherical morphology and micrometric size. Particles based on PLA/PEG and LSC significantly increased the apparent permeability coefficient (P_app) of PAs in Caco‐2 cells compared to the unencapsulated extract, indicating that these types of particles could be used as a delivery system for PA grape seed, allowing it to increase its bioavailability.     

Investigation of targeted pyrrolizidine alkaloids in traditional Chinese medicines and selected herbal teas sourced in Ireland using LC-ESI-MS/MS

Publications linking hepatotoxicity to the use of herbal preparations are escalating. Herbal teas, traditional Chinese medicines (TCMs) and dietary supplements have been shown to contain pyrrolizidine alkaloids (PAs). Acute PA toxicosis of the liver can result in sinusoidal-obstruction syndrome, also known as veno-occlusive disease (VOD). This paper describes a sensitive and robust method for the detection of targeted PAs and their N-oxides (PANOs) in herbal products (selected herbal teas and TCMs) sourced within Ireland. The sample preparation includes a simple acidic extraction with clean-up via solid-phase extraction (SPE). Sample extracts were accurately analysed by using LC-ESI-MS/MS applying for the first time a pentafluorophenyl (PFP) core-shell column to the chromatographic separation of PAs and PANOs. The method was validated for selectivity, taking into consideration matrix effects, specificity, linearity, precision and trueness. Limits of detection (LOD) and limits of quantitation (LOQ) were quantified for all PAs and PANOs ranging from 0.4 to 1.9 µg kg^?1 and from 1.3 to 6.3 µg kg^?1, respectively. In this study 10 PAs and four PANOs were targeted because they are commercially available as reference standards. Therefore, this study can only report the levels of these PAs and PANOs analysed in the herbal teas and TCMs. The results reported represent the minimum levels of PAs and PANOs present in the samples analysed; commercially available herbal teas (n = 18) and TCMs (n = 54). A total of 50% herbal teas and 78% Chinese medicines tested positive for one or more PAs and/or PANOs included within this study, ranging from 10 to 1733 and from 13 to 3668 µg kg^?1, respectively.     

Determination of Pyrrolizidine Alkaloids in Honey with Sensitive Gas Chromatography-Mass Spectrometry Method

Pyrrolizidine alkaloids (PAs) are secondary metabolites produced by plants as a chemical defense against herbivores. Plants containing PAs are widely distributed in almost all geographical regions posing a risk of honey contamination. To provide safety of honey and decrease the potential risk for the consumers, a sensitive method based on gas chromatography-mass spectrometry enabling determination of a content of 1,2-unsaturated PAs in honey was developed. Honey samples were purified on MCX cartridges, and PAs were eluted with a solvent mixture consisting of ethyl acetate, methanol, ammonia, and triethylamine. Subsequently, 1,2-unsaturated alkaloids were reduced to their common backbone structures and derivatized with heptafluorobutyric anhydride. The method was validated according to SANTE 2015. All received parameters are in consistence with the document requirements as recovery ranged from 73.1 to 93.6%. The repeatability and reproducibility were calculated as relative standard deviation and ranged from 3.9 to 8.6% and from 10.6 to 17.8%, respectively. The limit of quantification was determined as 1 μg kg^?1. Good linearity of the method was obtained with the coefficient of determination R ²?>?0.99. The method was applied to 40 Polish and 14 Asian honey sample analyses.     

Short communication: Antimicrobial effect of lactoferrin and its amidated and pepsin-digested derivatives against Salmonella Enteritidis and Pseudomonas fluorescens

The antimicrobial effect of bovine lactoferrin (LF) and its amidated and pepsin-digested derivatives, at concentrations varying from 0.25 to 20 mg/mL, against 3 Salmonella Enteritidis strains and 3 Pseudomonas fluorescens strains was investigated. Lactoferrin showed its maximum antimicrobial effect at 10 mg/mL against the 3 Salmonella strains, with reductions ranging from 1.3 to 2.0 log units, and the 3 Pseudomonas strains, with reductions ranging from 1.8 to 5.4 log units. In the case of amidated LF, the maximum effect on the 3 Salmonella strains was recorded at 0.25 mg/mL, with reductions in the range of 0.8 to 1.2 log units, whereas it was recorded at 1 mg/mL for the 3 Pseudomonas strains, with reductions in the range of 4.4 to 6.0 log units. Pepsin-digested LF showed its maximum antimicrobial effect at 1 mg/mL against the 3 Salmonella strains, with reductions ranging from 2.6 to 3.4 log units, and at 20 mg/mL against the 3 Pseudomonas strains, with reductions ranging from 4.5 to 5.4 log units. It is worth noting the pronounced effect (reductions exceeding 2.5 log units) of a low (1 mg/mL) concentration of pepsin-digested LF, which is naturally formed in the gastrointestinal tract, on Salmonella and Pseudomonas strains. A highly significant inverse correlation was found between capsule polysaccharide levels of bacterial strains and their lethality in the presence of different concentrations of amidated lactoferrin.     

Inter-laboratory comparison study for pyrrolizidine alkaloids in animal feed using spiked and incurred material

Pyrrolizidine alkaloids (PAs) are hepatotoxic metabolites produced by plants. PAs in animal feed can cause acute or chronic intoxications in animals and can be transferred to milk. An inter-laboratory comparison study among 12 laboratories, using their own methods of analysis, was conducted for the detection and quantification of PAs in animal feed. The participants were asked to quantify PAs in a blank test sample, a blank test sample to be spiked with a provided spiking mixture of seven PA standards, and a test sample contaminated with common groundsel (Senecio vulgaris). Ten of the participating laboratories used an LC-MS/MS method, one used an LC-ToF-MS method, and one used a GC-MS method. None of the laboratories reported false-negative samples, while two laboratories reported false-positive results in the blank sample. z-scores were calculated for each laboratory for seven PAs in test samples B and C. z-scores varied considerably between laboratories for the concentrations of the free bases and less for the N-oxides, probably due to the lower levels of the free bases as compared with the N-oxides in the contaminated feed. Questionable or unsatisfactory results for the z-scores were obtained for 8% of the cases for the spiked sample and for 12% of the incurred sample. Three laboratories scored consequently positive or negative results. No preferred method for quantification of PAs in feed could be identified within the methods used for this study due to the relatively small number of participants. It was concluded that this inter-laboratory study shows that the methods used for PA detection need further development for accurate estimation of PAs in contaminated feed.     

茶叶中吡咯里西啶生物碱检测技术、污染水平及健康风险研究进展

吡咯里西啶生物碱（pyrrolizidine alkaloids,PAs）是一类植物次级代谢产物,其中不饱和PAs及大部分PAs的氮氧化物具有多种毒性作用。PAs已被发现广泛存在于各类茶叶中,且成为导致我国茶叶出口受阻的主要污染物之一。本文综述了茶叶中PAs检测的前处理方法和色谱-质谱检测技术的发展,并展望其发展趋势;总结了不同国家茶叶PAs污染水平;分析了茶叶中PAs种类、不同茶叶的PAs差异以及茶叶与其他食品或农产品中PAs差异;解析了茶叶中PAs对人体的健康风险。我国茶叶PAs污染水平监测与风险评估研究相关数据处于缺失状态,急需开展茶叶中PAs污染来源与风险评估研究。