Search

Effects of two foods with bioactive constituents (black tea brew, BTB and grape seed powder, GSP) on lipid digestibility was studied. Lipolysis inhibitory effect of these foods was examined using two test foods (cream and baked beef) with highly different fatty acid (FA) composition. Digestion simulations were performed either using both gastric and pancreatic lipase, or only with pancreatic lipase according to the Infogest protocol. Lipid digestibility was assessed based on the bioaccessible FAs. Results showed the triacylglycerols containing short- and medium-chain FAs (SCFA and MCFA) are non-preferred substrates for pancreatic lipase; however, this is not characteristic for GL. Our findings suggest that both GSP and BTB primarily affect the lipolysis of SCFAs and MCFAs, because the dispreference of pancreatic lipase towards these substrates was further enhanced as a result of co-digestion. Interestingly, GSP and BTB similarly resulted in significant decrease in lipolysis for cream (containing milk fat having a diverse FA profile), whereas they were ineffective in influencing the digestion of beef fat, having simpler FA profile. It highlights that the characteristics of the dietary fat source of a meal can be a key determinant on the observed extent of lipolysis when co-digested with foods with bioactive constituents.

Sweeteners of natural sources, such as sugar alcohols, are in the centre of growing interest. Their impact on the phytochemicals, antioxidant and sensory properties of blackberry jams were investigated during a 9-months storage period. Measurements on jams prepared with different sugars and sugar alcohols (sucrose, fructose, xylitol and erythritol) were performed at the date of preparation and in the 1st, 3rd, 6th, 9th month.Total polyphenol content, individual polyphenols, antioxidant properties, anthocyanin content and CIE L*a*b* colour were determined. Sensory profiles were determined by a trained panel. Sensory attributes were compared by the results of the electronic tongue and nose.Sweeteners had a significant impact on physicochemical properties and sensory attributes. Storage time also affected the sensory and compositional properties of jams. Changes in antioxidant properties did not follow a clear trend during the storage period, and antioxidant capacity was not affected significantly by the sweetening agent, but showed a significant decline from the 6th month. A positive effect of xylitol was observed in terms of a low degradation rate of anthocyanins, while their decomposition was the fastest in the fructose-containing preparation. Jams sweetened with erythritol reached significantly lower values for some sensory attributes (blackberry flavour and general taste intensity), however, they showed more intense red colour. Multiple factor analysis enabled the identification of the effect of sweetener and storage time on the pattern of the sensory data matrix. Classification according to individual sweeteners was performed successfully by the electronic tongue, but not by electronic nose.

The nutritional relevance of food compositional data could be improved by taking the bioaccessibility of these constituents into account. A lack of routine methods to assess the bioaccessibility of fatty acids (FAs) in food is one of the limiting factors of doing so. An analytical protocol is proposed for routine assessment of the extent of lipolysis via in vitro digestion simulation methods in food products. The established method provides specific information on each FA individually. Steps of the protocol including the Bligh and Dyer chloroform/methanol/water extraction of esterified and free FAs from in vitro digesta, methyl ester derivatization, and GC-FID analysis were specifically tailored to help routine work and were harmonized with the Infogest in vitro digestion simulation protocol (both v1.0 and v2.0). The method was applied to assess the degree of FA-specific lipolysis in a baked fish (carp) meal and the results showed that the FA composition of the original food significantly differed from that of the distribution of FFAs in the digesta. The use of gastric lipase (in Infogest v2.0 protocol) increased total FA release by 9.5% and its specific impact on palmitic acid was the most prominent.

Prediction of retention times (RTs) is increasingly considered in untargeted metabolomics to complement MS/MS matching for annotation of unidentified peaks. We tested the performance of PredRet (http://predret.org/) to predict RTs for plant food bioactive metabolites in a data sharing initiative containing entry sets of 29–103 compounds (totalling 467 compounds, >30 families) across 24 chromatographic systems (CSs). Between 27 and 667 predictions were obtained with a median prediction error of 0.03–0.76 min and interval width of 0.33–8.78 min. An external validation test of eight CSs showed high prediction accuracy. RT prediction was dependent on shape and type of LC gradient, and number of commonly measured compounds. Our study highlights PredRet’s accuracy and ability to transpose RT data acquired from one CS to another CS. We recommend extensive RT data sharing in PredRet by the community interested in plant food bioactive metabolites to achieve a powerful community-driven open-access tool for metabolomics annotation., The authors acknowledge networking support by the European Cooperation in Science and Technology (COST) Action FA 1403 POSITIVe (Interindividual Variation in Response to Consumption of Plant Food Bioactives and Determinants Involved). Dorrain Low has received the support of EU H2020 in the framework of the FP7-Marie Curie-COFUND People Programme, through award of the AgreenSkills+ Fellowship (grant number 609398) and Nanyang Technological University, Singapore, through award of the Presidential Postdoctoral Fellowship (grant number 001991-00001). The MetaboHUB French infrastructure has supported the INRAE platform (PFEM, MetaboHUB-Clermont) involved in this work (grant number ANR-INBS-0010). Kateřina Valentová and Lucie Petrásková acknowledge financial support from the Czech Science Foundation (grant number 19-00043S). The Faculty of Pharmacy of Lisbon University acknowledges FUNDAÇÃO PARA A CIÊNCIA E TECNOLOGIA and PORTUGAL 2020 for financial support of the Portuguese Mass Spectrometry Network (grant number LISBOA-01-0145-FEDER-402-022125). Kati Hanhineva and Ville Koistinen have received funding from the Academy of Finland (grant numbers 277986 and 312550), Lantmännen Foundation and EU H2020 FP7-Marie Curie-COFUND MoRE Programme (grant number 754412). Biocenter Finland has financially supported the LC-MS metabolomics unit of University of Eastern Finland. Cristina Andres-Lacueva and Raúl González-Domínguez thank CIBERFES and ISCIII projects AC19/00111 and AC19/00096 (co-funded by FEDER Program from EU, “A way to make Europe”), Generalitat de Catalunya’s Agency AGAUR (grant number 2017SGR1546), “Juan de la Cierva” program from MINECO (grant number IJC2019-041867-I) and ICREA Academia award 2018. Francisco A. Tomás-Barberán has received financial support from the Spanish National Research program (grant numbers AGL-2015-73107-EXP/AEI, CSIC 201870E014) and Fundación Seneca Región de Murcia (grant number 19900/GERM/15). Gesine Schmidt acknowledges support through the Norwegian Agriculture and Food Industry Research Funds (grant number 262300). Lars Dragsted and Jan Stanstrup thank the Carlsberg Foundation for a Semper Ardens grant to support this work. László Abrankó acknowledges the Hungarian Academy of Sciences for the János Bolyai Scholarship, and support of the EU and ESF co-financed project of SZIU (grant number EFOP-3.6.3-VEKOP-16-2017-00005). Sonia de Pascual-Teresa thanks the Spanish MINECO for financial support (grant number AGL2016-76832-R). Dilip K. Rai acknowledges Teagasc for the financial support through the Walsh Fellowship (grant number 2016038).

Liquid milk protein concentrate with different beneficial values was prepared by membrane filtration and enzymatic modification of proteins in a sequential way. In the first step, milk protein concentrate was produced from ultra-heat-treated skimmed milk by removing milk serum as permeate. A tubular ceramic-made membrane with filtration area 5 &times, 10&minus, 3 m2 and pore size 5 nm, placed in a cross-flow membrane house, was adopted. Superior operational strategy in filtration process was herein: trans-membrane pressure 3 bar, retention flow rate 100 L&middot, h&minus, 1, and implementation of a static turbulence promoter within the tubular membrane. Milk with concentrated proteins from retentate side was treated with the different concentrations of trypsin, ranging from 0.008&ndash, 0.064 g&middot, L&minus, 1 in individual batch-mode operations at temperature 40 &deg, C for 10 min. Subsequently, inactivation of trypsin in reaction was done at a temperature of 70 &deg, C for 30 min of incubation. Antioxidant capacity in enzyme-treated liquid milk protein concentrate was measured with the Ferric reducing ability of plasma assay. The reduction of angiotensin converting enzyme activity by enzyme-treated liquid milk protein concentrate was measured with substrate (Abz-FRK(Dnp)-P) and recombinant angiotensin converting enzyme. The antibacterial activity of enzyme-treated liquid milk protein concentrate towards Bacillus cereus and Staphylococcus aureus was tested. Antioxidant capacity, anti-angiotensin converting enzyme activity, and antibacterial activity were increased with the increase of trypsin concentration in proteolytic reaction. Immune-reactive proteins in enzyme-treated liquid milk protein concentrate were identified with clinically proved milk positive pooled human serum and peroxidase-labelled anti-human Immunoglobulin E. The reduction of allergenicity in milk protein concentrate was enzyme dose-dependent.

Tart cherries have been reported to exert potential health benefits attributed to their specific and abundant polyphenol content. However, there is a need to study the impact and fate of tart cherries polyphenols in the gut microbiota. Here, tart cherries, pure polyphenols (and apricots) were submitted to in vitro bacterial fermentation assays and assessed through 16S rRNA gene sequence sequencing and metabolomics. A short-term (5 days, 8 oz. daily) human dietary intervention study was also conducted for microbiota analyses. Tart cherry concentrate juices were found to contain expected abundances of anthocyanins (cyanidin-glycosylrutinoside) and flavonoids (quercetin-rutinoside) and high amounts of chlorogenic and neochlorogenic acids. Targeted metabolomics confirmed that gut microbes were able to degrade those polyphenols mainly to 4-hydroxyphenylpropionic acids and to lower amounts of epicatechin and 4-hydroxybenzoic acids. Tart cherries were found to induce a large increase of Bacteroides in vitro, likely due to the input of polysaccharides, but prebiotic effect was also suggested by Bifidobacterium increase from chlorogenic acid. In the human study, two distinct and inverse responses to tart cherry consumption were associated with initial levels of Bacteroides. High-Bacteroides individuals responded with a decrease in Bacteroides and Bifidobacterium, and an increase of Lachnospiraceae, Ruminococcus and Collinsella. Low-Bacteroides individuals responded with an increase in Bacteroides or Prevotella and Bifidobacterium, and a decrease of Lachnospiraceae, Ruminococcus and Collinsella. These data confirm that gut microbiota metabolism, in particular the potential existence of different metabotypes, needs to be considered in studies attempting to link tart cherries consumption and health.

Fatty acyl-Coenzyme A species (acyl-CoAs) are key biomarkers in studies focusing on cellular energy metabolism. Existing analytical approaches are unable to simultaneously detect the full range of short-, medium-, and long-chain acyl-CoAs, while chromatographic limitations encountered in the analysis of limited amounts of biological samples are an often overlooked problem. We report the systematic development of a UHPLC-ESI-MS/MS method which incorporates reversed phase (RP) and hydrophilic interaction liquid chromatography (HILIC) separations in series, in an automated mode. The protocol outlined encompasses quantification of acyl-CoAs of varying hydrophobicity from C2 to C20 with recoveries in the range of 90-111 % and limit of detection (LOD) 1-5 fmol, which is substantially lower than previously published methods. We demonstrate that the poor chromatographic performance and signal losses in MS detection, typically observed for phosphorylated organic molecules, can be avoided by the incorporation of a 0.1% phosphoric acid wash step between injections. The methodological approach presented here permits a highly reliable, sensitive and precise analysis of small amounts of tissues and cell samples as demonstrated in mouse liver, human hepatic (HepG2) and skeletal muscle (LHCNM2) cells. The considerable improvements discussed pave the way for acyl-CoAs to be incorporated in routine targeted lipid biomarker profile studies.

The history of the acyl-quinic acids is briefly reviewed, the merits and limitations of the various nomenclature systems applicable are critically compared, and their limitations are highlighted, in particular their inability to provide an unambiguous description of all quinic acid enantiomers and diastereoisomers and associated acyl-quinic acids. Recommendations are made for a nomenclature system that in combination with IUPAC numbering achieves this objective. A comprehensive set of structures for the quinic acid enantiomers and diastereoisomers is presented. The Supporting Information provides an explanation of trivial names and a decision tree to determine which quinic acid isomer a structure represents.

Hydroxycinnamoylquinic acids (HCQAs) are a major class of phenolic plant secondary metabolites, belonging to the chlorogenic acid family. Various health-beneficial properties of HCQAs have been shown, which has drawn interest for HCQA profiling in plants of human consumption. However, this task remains challenging, because several isomeric HCQAs can be present in the sample with identical molecular formulae and the limited availability of reference standards poses additional challenges to their identification. In the present work, a high performance liquid chromatography–electrospray ionization–quadrupole time-of-flight–mass spectrometry (HPLC-ESI-Q/TOF-MS) method accompanied with an effective data filtering protocol is presented, which is shown to be suitable for the identification of HCQAs in plant materials in a non-targeted manner. Both collision-induced dissociation (CID) fragmentation performed in a collision cell and in-source (CID) fragmentation were used to produce accurate mass fragments. It was shown that fragmentation characteristics required for identification of regio-isomers of HCQAs can be achieved with in-source CID fragmentation, enabling the use of a single-stage MS system with in-source fragmentation for convincing identification of HCQAs. Based on a thorough validation of identified HCQA compounds using coffee bean extracts as reference samples, comprehensive profiling of HCQAs in two apricot (Prunus armeniaca L.) genotypes (‘Preventa’ and ‘Gonci magyarkajszi’) was carried out for the first time and the following 10 HCQAs were shown to be present in apricot fruit: 3-caffeoylquinic acid (CQA), cis-3-CQA, 4-CQA, 5-CQA, cis-5-CQA, 3,5-diCQA, 3-p-coumaroylquinic acid (pCoQA), 4-pCoQA, 3-feruloylquinic acid (FQA) and cis-3-FQA. Copyright © 2016 John Wiley & Sons, Ltd. Highlights An HPLC-ESI-Q/TOF-MS method suitable for the identification of hydroxycinnamoyilquinic acids (HCQAs) in plant material in a non-targeted manner was developed. Single-stage, high-resolution MS system with in-source fragmentation was shown to be suitable for convincing identification of HCQAs. Comprehensive profiling of HCQAs in two apricot (Prunus armeniaca L.) genotypes was carried out for the first time. Copyright © 2016 John Wiley & Sons, Ltd.

Ground ivy (Glechoma hederacea L.) is one of the prosperous plants for the food-industry as natural antioxidant. This fact led us to examine the chemical diversity of six ground ivy populations situated in different natural habitats and to analyse the effect of the harvesting time. Total phenolic content, chlorogenic acid, and rutin content, as well as the antioxidant capacity showed significant differences due to the harvest time. The highest total phenol content (115 mg g–1 GAE) and the strongest antioxidant activity (53.3 mg g–1 AAE) were measured in the population originated from Budapest (GLE 6), harvested in July. The highest chlorogenic acid (357 mg/100 g) and rutin (950 mg/100 g) contents were detected in the July harvested samples from the Soroksar Botanical Garden population (GLE 1). According to our results, the collection time has significant effect on the total phenolic content – first of all on the chlorogenic acid and rutin accumulation levels of ground ivy, while the influence of the habit...

The aim of this study is to examine the effect of arbuscular mycorrhizal fungi (AMF) colonization on biomass, polyphenol profile, and content of economically important herbs. A pot experiment was performed with marjoram, lemon balm, and marigold applying a commercially available AMF mixture for inoculation. Major polyphenols were identified using HPLC-UV-ESI-qTOFMS on the basis of their UV-vis and mass spectral characteristics, and selected ones were quantified. We showed that AMF can provide different services for each herb. Marjoram had the highest level of fungal colonization (82 M%) followed by lemon balm (62 M%) and marigold (17 M%). AMF inoculation significantly increased the biomass of marjoram (1.5-fold), the number of marigold flowers (1.2-fold), and the yield of rosmarinic acid and lithospermic acid isomers of marjoram (1.5-fold) and lemon balm (1.2-fold). Therefore, the quantity and quality of plant material could be improved by the application of optimized AMF inoculum.

Summary Elderberry concentrate is widely used for food colouring. Its production includes several processing steps, which influence pigment composition, thus affecting the quality of final product. In this study, qualitative and quantitative changes in anthocyanin components were followed during the production of elderberry juice concentrate using Haschberg and Samocco varieties. Juice concentration resulted in more than five-fold concentration increase in anthocyanins; however, clarification, filtration and evaporation steps caused slight but continuous decrease in anthocyanins relative to the dry matter of the product. Pectolytic enzymatic treatment was found to be one of the most critical processing steps that affects anthocyanin yield during production, and this effect on yield can be variety-dependent. Additionally, two condensed dimeric anthocyanin species generated after heat treatment are reported for the first time to be present in elderberry juice concentrate. Our results suggest that cyanidin-based anthocyanin conjugates with more complex saccharide moieties are more robust during production.

Transient hyperglycaemia is a risk factor for type 2 diabetes and endothelial dysfunction, especially in subjects with impaired glucose tolerance. Nutritional interventions and strategies for controlling postprandial overshoot of blood sugars are considered key in preventing progress to the disease state. We have identified apigenin-7-O-glucoside, apigenin, and (Z) and (E)−2-hydroxy-4-methoxycinnamic acid glucosides as the active (poly)phenols in Chamomile (Matricaria recutita) able to modulate carbohydrate digestion and absorption in vitro as assessed by inhibition of α-amylase and maltase activities. The latter two compounds previously mistakenly identified as ferulic acid hexosides were purified and characterised and studied for their contribution to the overall bioactivity of chamomile. Molecular docking studies revealed that apigenin and cinnamic acids present totally different poses in the active site of human α-amylase. In differentiated Caco-2/TC7 cell monolayers, apigenin-7-O-glucoside and apigenin strongly inhibited D-[U-14C]-glucose and D-[U-14C]-sucrose transport, and less effectively D-[U-14C]-fructose transport. Inhibition of D-[U-14C]-glucose transport by apigenin was stronger under Na+-depleted conditions, suggesting interaction with the GLUT2 transporter. Competitive binding studies with molecular probes indicate apigenin interacts primarily at the exofacial-binding site of GLUT2. Taken together, the individual components of Chamomile are promising agents for regulating carbohydrate digestion and sugar absorption at the site of the gastrointestinal tract.

Bioactive compounds present in plant-based foods, and their metabolites derived from gut microbiota and endogenous metabolism, represent thousands of chemical structures of potential interest for human nutrition and health. State-of-the-art analytical methodologies, including untargeted metabolomics based on high-resolution mass spectrometry, are required for the profiling of these compounds in complex matrices, including plant food materials and biofluids. The aim of this project was to compare the analytical coverage of untargeted metabolomics methods independently developed and employed in various European platforms. In total, 56 chemical standards representing the most common classes of bioactive compounds spread over a wide chemical space were selected and analyzed by the participating platforms (n = 13) using their preferred untargeted method. The results were used to define analytical criteria for a successful analysis of plant food bioactives. Furthermore, they will serve as a basis for an optimized consensus method., This research was funded by the Generalitat de Catalunya’s Agency AGAUR (grant number 2017 SGR 1546), the Spanish MINECO´s “Juan de la Cierva” programme (grant number FJCI-2015-26590), Spanish National Programme for Research Aimed at the Challenges of Society (AGL2016-76832-R), the Czech Science Foundation (grant number 18-00132S), the European Structural and Investment Funds (grant agreement No. VEKOP-2.3.3-15-2017-00022), and Fundação para a Ciência e Tecnologia (grant number LISBOA-01-0145-FEDER-402-022125). Networking support was provided by the COST Action FA1403 POSITIVe (Inter-individual variation in response to consumption of plant food bioactives and determinants involved), supported by COST (European Cooperation in Science and Technology).

Summary Profiling and quantitative analysis of anthocyanins in five elderberry (Sambucus nigra L.) varieties, namely ‘Haschberg’, ‘Samocco’, ‘Samyl’, ‘Samident’ and ‘Sampo’, were performed in six different maturity stages from two consecutive years (2012 and 2013) and from two growing areas in Hungary. Cyanidin-3-O-sambubioside-5-O-glucoside, cyanidin-3-O-sambubioside and cyanidin-3-O-glucoside were found and identified by HPLC-Q/TOF-MS as major anthocyanins and were quantified by HPLC-UV/Vis. In optimum maturity stage, ‘Samocco’ showed the highest anthocyanin content with an average of 1237 mg per 100 g dry weight in both growing areas and vintages. The dominant anthocyanin component of Samocco variety was cyanidin-3-O-sambubioside, which is according to literature more stable against technology processing than cyanidin-3-O-glucoside found in the other four investigated elderberry varieties in the highest concentration. ‘Samocco’, if grown under the climatic conditions of the Carpathian basin, might be a promising alternative variety for growing as raw material for natural food colourant processing industry.

Elderberry (Sambucus nigra subsp. nigra) is a very popular crop in Europe and its importance is increasing as a distinguished source of natural food colorants. The amount of natural pigments in fruits depends on several factors such as the plant cultivar, the geographical origin, the climatic conditions, and the time of harvest. Cyanidin-3-O-sambubioside-5-O-glucoside, cyanidin-3-O-sambubioside and cyanidin-3-O-glucoside were identified in elderberry fruit by liquid chromatography-mass spectrometry (LC-MS), and quantified by high performance liquid chromatography (HPLC-Vis). In a fruit maturity study, ‘Samocco’ had the highest water soluble solid content compared with two other elderberry cultivars grown at two locations. Anthocyanin profile and content also differed by cultivar. With ‘Samocco’, cyanidin-3-O-sambubioside accounted for more than 50% of all analysed anthocyanins, but the amount of this pigment was less in the other cultivars. ‘Samocco’ had higher levels of total anthocyanins compared with most other cultivars at the two locations. Anthocyanin content was influenced by change of pH value and by heat treatment. After heat treatment, the highest anthocyanin concentration was observed at pH 3.5 with ‘Haschberg’, and at pH 3.0 with ‘Samocco’. Total anthocyanin content was decreased 7% by heat treatment with ‘Haschberg’ and by 25% with ‘Samocco’ at the original pH value of 4.0.

The isoflavone genistein on the contrary to its well-established health-beneficial effects is not a major component of the Western diet, since soy consumption, considered as the main dietary source of genistein, in these populations is low. Genistein compounds in twelve commercial sour cherry (Prunus cerasus L.) cultivars grown in Hungary were studied. High performance liquid chromatography coupled to electrospray ionisation quadrupole/time-of-flight mass spectrometry (HPLC-ESI-qToF-MS) was used for screening and confirmatory analyses. Genistin and genistein were found in 'Pipacs1', 'Kántorjánosi', 'Debreceni bőtermő' and 'Éva', which are native cultivars to Hungary. Genistein content of the latter three were in the range of 0.4-0.6, while in 'Pipacs1' in total 4.4 mg genistein compounds were measured expressed as aglycone equivalents per 100g of fresh fruit flesh. These cultivars may play important role as complementary genistein sources in the Western diet. Especially 'Pipacs 1', may be best utilised in functional food products.

In fields such as food and nutrition science or plant physiology, interest in untargeted profiling of flavonoids continues to expand. The group of flavonoids encompasses several thousands of chemically distinguishable compounds, among which are a number of isobaric compounds with the same elemental composition. Thus, the mass spectrometric identification of these compounds is challenging, especially when reference standards are not available to support their identification. Many different types of isomers of flavonoid glycoconjugates are known, i.e. compounds that differ in their glycosylation position, glycan sequence or type of interglycosidic linkage. This work focuses on the mass spectrometric identification of flavonoid glycoconjugate isomers possessing the same glycan mass and differing only in their aglycone core. A non-targeted HPLC-ESI-MS/MS profiling method using a triple quadrupole MS is presented herein, which utilizes in-source fragmentation and a pseudo-MS(3) approach for the selective analysis of flavonoid glycoconjugates with isomeric/isobaric aglycones. A selective MRM-based identification of the in-source formed isobaric aglycone fragments was established. Additionally, utilizing the precursor scanning capability of the employed triple quadrupole instrument, the developed method enabled the determination of the molecular weight of the studied intact flavonoid glycoconjugate. The versatility of the method was proven with various types of flavonoid aglycones, i.e. anthocyanins, flavonols, flavones, flavanones and isoflavones, along with their representative glycoconjugates. The developed method was also successfully applied to a commercially available sour cherry sample, in which 16 different glycoconjugates of pelargonidin, genistein, cyanidin, kaempferol and quercetin could be tentatively identified, including a number of compounds containing isomeric/isobaric aglycones.

The effect of temperature (6, 15 and 30 °C) during ageing on the colour, phytochemical composition and bioactivity of roselle wine was investigated over 12 months. At the end of ageing, wines stored at 6 °C had the highest colour density and lowest polymeric anthocyanins. The initial concentration of most of the individual phenolic compounds decreased during ageing, with reduction of monomeric anthocyanins contributing to the formation of anthocyanin-derivatives (pyranoanthocyanins), eight of which were identified tentatively and reported here for the first time in roselle wine. The decrease in individual phenolic compounds did not affect inhibition of α-glucosidase (maltase) activity, which remained relatively low but stable throughout ageing. Diethyl succinate was the only volatile clearly influenced by ageing temperature, with the most pronounced effect at 30 °C (∼256 fold increase). In summary, the final concentrations of anthocyanins and diethyl succinate were the major compounds influenced by ageing temperature.

Dietary fiber-derived short-chain fatty acids (SCFA) and phenolics produced by the gut microbiome have multiple effects on health. We have tested the hypothesis that long-term exposure to physiological concentrations of SCFA can affect the transport and metabolism of (poly)phenols by the intestinal epithelium using the Caco-2 cell model. Metabolites and conjugates of hesperetin (HT) and ferulic acid (FA), gut-derived from dietary hesperidin and chlorogenic acid, respectively, were quantified by LC-MS with authentic standards following transport across differentiated cell monolayers. Changes in metabolite levels were correlated with effects on mRNA and protein expression of key enzymes and transporters. Propionate and butyrate increased both FA transport and rate of appearance of FA glucuronide apically and basolaterally, linked to an induction of MCT1. Propionate was the only SCFA that augmented the rate of formation of basolateral FA sulfate conjugates, possibly via basolateral transporter up-regulation. In addition, propionate enhanced the formation of HT glucuronide conjugates and increased HT sulfate efflux toward the basolateral compartment. Acetate treatment amplified transepithelial transport of FA in the apical to basolateral direction, associated with lower levels of MCT1 protein expression. Metabolism and transport of both HT and FA were curtailed by the organic acid lactate owing to a reduction of UGT1A1 protein levels. Our data indicate a direct interaction between microbiota-derived metabolites of (poly)phenols and SCFA through modulation of transporters and conjugating enzymes and increase our understanding of how dietary fiber, via the microbiome, may affect and enhance uptake of bioactive molecules.

Vitamin D2 is essential for the human body and mushrooms are proved to be one of the best sources of it. In our experiment biologically active, pre-harvest white and cream type button mushrooms and oyster mushroom were treated with UVB light in order to increase their vitamin D2 level, by transforming their natural ergosterol content into vitamin D2. UVB lamps (operating on 312 nm) and six time periods of irradiation (15 to 90 min) were used. After three consecutive days of treatments the yield were measured and samples were taken for vitamin D2 analysis. Data showed considerable increase in vitamin D2 levels at every time period in each cultivar.

The aim of this study was the comparison of different sample preparation methods for multielemental ICP-MS analysis of wine samples. All investigated methods were previously published for the same purpose, but the comprehensive comparison was carried out in the frame of this study for the first time. Five wine samples of different residual sugar content (2 dry, 2 sweet and one extra fine quality dessert wine) were used from Hungarian wine districts. Due to the lack of certified reference materials method validation and cross-checking was carried out by using two different types of inductively coupled plasma mass spectrometers (ICP-MS), namely a quadrupole based ICP-MS equipped with a collision cell (ICP-CC-MS) and a high resolution sector field ICP-MS (ICP-SF-MS). The measured elements were Li, Mg, B, Al, Cu, Rb, Sr, Zr, Cd, Ba, Tl, Pb, Bi, U, Cr, Fe, Mn, Co, Ni, Zn, As, Mo and Ga.Sample preparation procedures were based on sample dilution, microwave digestion using nitric acid (in combination with and without evaporation prior to ICP-MS analysis) and a mixture of nitric acid and hydrogen peroxide. The investigation resulted that the optimal digestion method for the above-mentioned purpose is microwave digestion using nitric acid without evaporation. The advantages and disadvantages of the investigated methods are critically discussed.

In recent years, the detection and characterization of relevant pesticide metabolites in food is an important task in order to evaluate their formation, kinetics, stability, and toxicity. In this article, a methodology for the systematic screening of pesticides and their main metabolites in fruit and vegetable samples is described, using LC-HRMS and accurate-mass database search of parent compounds and their diagnostic fragment ions. The approach is based on (i) search for parent pesticide molecules; (ii) search for their metabolites in the positive samples, assuming common fragmentation pathways between the metabolites and parent pesticide molecules; and (iii) search for pesticide conjugates using the data from both parent species and diagnostic fragment ions. An accurate-mass database was constructed consisting of 1396 compounds (850 parent compounds, 447 fragment ions and 99 metabolites). The screening process was performed by the software in an automated fashion. The proposed methodology was evaluated with 29 incurred samples and the output obtained was compared to standard pesticide testing methods (targeted LC-MS/MS). Examples on the application of the proposed approach are shown, including the detection of several pesticide glycosides derivatives, which were found with significantly relevant intensities. Glucose-conjugated forms of parent compounds (e.g., fenhexamid-O-glucoside) and those of metabolites (e.g., despropyl-iprodione-N-glycoside) were detected. Facing the lack of standards for glycosylated pesticides, the study was completed with the synthesis of fenhexamid-O-glucoside for quantification purposes. In some cases the pesticide derivatives were found in a relatively high ratio, drawing the attention to these kinds of metabolites and showing that they should not be neglected in multi-residue methods. The global coverage obtained on the 29 analyzed samples showed the usefulness and benefits of the proposed approach and highlights the practical benefit obtained when the so-called screening methods are used as a complementary tool to standard targeted LC-MS/MS methods.

In this work, a systematic comprehensive screening procedure has been proposed for the detection of multiclass flavonoid derivatives by liquid chromatography high-resolution mass spectrometry (LC-HRMS). The procedure is based on the combined use of accurate mass measurements and in-source fragmentation obtained with a liquid chromatography time-of-flight mass spectrometry instrument. The method relies on automated screening of selected diagnostic ions based on an exact mass database. The included diagnostic ions represent theoretical combinations of aglycones and typical glycan part constituents of flavonoid derivatives (i.e., various saccharide units and acyl moieties). The proposed identification protocol is following a systematic evaluation of the obtained positive hits from the diagnostic ions database according to a "bottom-up" approach that is thoroughly discussed. The main benefit of the proposed bottom-up protocol resides in the fact that untargeted flavonoid derivatives can be detected and tentatively identified without the need for any preliminary knowledge on the sought compound. In addition to information on the nature of the (1) aglycone and the (2) glycan part, further indication of (3) sugar unit distribution and information on (4) the type of the glycosidic bonds can also be attained. Selected examples of plant extracts demonstrate the potential of the proposed LC-HRMS approach for the systematic screening of flavonoids. A broad variety of compounds were tentatively identified including both anthocyanins and non-anthocyanin flavonoids having various glycan moieties such as mono-, di-, and triglycosides with varying distributions and linkage types of carbohydrate moieties (O-glycosides, C-glycosides, O,C-glycosides).

A rapid LC–MS/MS method for quantification of phenols and polyphenols in authentic wine samples with unrivalled sensitivity was developed. Excellent limits of detection in the low μg L −1 range were achieved. Reversed phase HPLC employing sub-2 μm particles as stationary phase allowed high-throughput analysis with analysis times of 10 min for 11 compounds. The phenolic pattern was assessed in 97 authentic wine samples (stored under identical conditions without further treatment or modification) comprising eleven geographical Austrian regions, six grape varieties and five vintages. Canonical discriminant analysis was applied to the data set showing the suitability of the (poly)phenolic spectrum for classification of the wine samples. The method allowed a geographic discrimination of several grape varieties and a grape variety based discrimination of four regions. As a novel finding excellent (poly)phenol-based differentiation of the five investigated vintages (2003–2007) was achieved.

An LC–UV screening method for simultaneous determination of ascorbic acid (C), and the free forms of thiamine (B1) riboflavin (B2), niacin (B3), pyridoxine (B6) in enriched food products was developed and validated. The chromatographic separation was accomplished within 18 min using a gradient of water with 0.1% formic acid (pH 2.5) and methanol with 0.1% formic acid on a C18 reverse phase column (5 μm, 150 × 3.2 mm) while detection was performed at two wavelengths (266 and 290 nm). Sample preparation was based on an extraction method originally developed for vitamin C. This procedure besides extracting vitamin C was extended to the extraction of the free forms of vitamins B1, B2, B3, B6 and B9. The developed analytical method was successfully applied for the simultaneous determination of the vitamin C content along with the free vitamin B forms of three different enriched food products.

Vitamins form a heterogeneous chemical group having different stability. In foodstuffs some of them might be bound to matrix components. In the case of vitamin supplemented food products, since the vitamins are not strongly embedded in the matrix a general extraction method could be fit for purpose. The aim of this study was the simultaneous determination of the most common water-soluble vitamins, i.e. ascorbic acid (C), riboflavin (B 2 ), niacin (B 3 ), pyridoxine (B 6 ), folic acid (B 9 ) in enriched food products. Sample preparation based on the European Standard (CEN, 2003) was optimised for further LC-MS compatible chromatography. The separation of the vitamins was achieved by reversed-phase liquid chromatography. Detection was carried out with a photodiode array detector at four different wavelengths. The chromatographic method and the sample preparation were successfully applied for vitamin-enriched cereal, instant cacao powder and fruit juice samples.

Summary Sour cherry (Prunus cerasus L.) cultivars native to Hungary and some of them grown worldwide, display great phenotypic variability. Eleven sour cherry cultivars were analysed for their main quality attributes including fruit weight, soluble solid content, acidity, pH, specific sugars (glucose and fructose), antioxidant capacity as well as total polyphenolic (TPC), total anthocyanin (TMAC) and vitamin C contents. Results showed wide variation in both fruit quality and antioxidant parameters of Hungarian sour cherries. The anthocyanin contents varied from 11.3 to 93.5 mg/100 g. An amarelle-type cultivar, ‘Pipacs 1’ showed the highest antioxidant capacity (21.85 mmol AA L−1), TPC (49.04 mg GA L−1) and vitamin C (8.98 mg/100 g) content. Key enzymes in anthocyanin biosynthesis were expressed in both yellow-coloured flesh and red skin of ‘Pipacs 1’ fruits at all ripening stages. The detected diversity presents a choice that can satisfy different consumer preferences, and meet specific nutritional requirements.

A three-step HPLC–ESI-MS/MS procedure is designed for screening and identification of non-target flavonoid derivatives of selected flavonoid aglycones. In this method the five commonly appearing aglycones (apigenin, luteolin, myricetin, naringenin and quercetin) were selected. The method consists of three individual mass spectrometric experiments of which the first two were implemented within a single chromatographic acquisition. The third step was carried out during a replicate chromatographic run using the same RP-HPLC conditions. The first step, a multiple reaction monitoring (MRM) scan of the aglycones was performed to define the number of derivatives relating to the selected aglycones. For this purpose the characteristic aglycone parts of the unknowns were used as specific tags of the molecules, which were generated as in-source fragments. Secondly, a full scan MS experiment is performed to identify the masses of the potential derivatives of the selected aglycones. Finally, the third step had the capability to confirm the supposed derivatives. The developed method was applied to a commercially available black currant juice to demonstrate its capability to detect and identify various flavonoid glycosides without any preliminary information about their presence in the sample. As a result 13 compounds were detected and identified in total. Namely, 3 different myricetin glycosides and the myricetin aglycone 2 luteolin glycosides plus the aglycone and 3 quercetin glycosides plus the aglycone could be identified from the tested black currant sample. In the case of apigenin and naringenin only the aglycones could be detected.

The optimization and application of gas chromatograph coupled with inductively coupled plasma mass spectrometer (GC–ICPMS) (equipped with a commercially available interface) for the speciation of butyltin compounds in freshwater origin sediment and mussel samples is described. Optimization focused on the system parameters that have the greatest effect on signal intensity such as plasma power, ion lenses and make up gas flow (in the interface). Xenon (Xe) containing argon gas (Ar) was applied as tuning gas providing continuous Xe signal for the optimization of system parameters. It was found that plasma power and make up gas are interrelated variables and provide a set of paired optimum values at each power settings. The absolute optimum values obtained at 800 W plasma power and 1.2 L min−1 make up gas flow rate when 7 mm sample depth was adjusted. The optimum settings obtained were then checked by means of a test solution (tetraethyltin dissolved in hexane). Same optimum conditions were found when tin (Sn) transient signals were monitored. Detection limits were calculated for the three species using the optimized system parameters. Detection limits are the following: for monobutyltin (MBT) 5.6 ng Sn kg−1, for dibutyltin (DBT) 6.6 ng Sn kg−1 and for tributyltin (TBT) 3.4 ng Sn kg−1 obtained. Determination of the butyltin compounds were carried out by means of species-specific isotope dilution analysis. The spike solution contained all species investigated but with altered isotopic composition. Each species were enriched in their 119Sn isotope. Concentrations found in Hungarian freshwater origin mussel and sediment samples ranged between 19 and 39 ng g−1for MBT, between 1.2 and 6.3 ng g−1 for DBT and between 1.2 and 3.2 ng g−1 for TBT indicated as Sn in dry weight. Validation of the method was done by means of certified reference materials (BCR CRM 646 and 477). Good agreement was found between certified and experimental values. Normalized deviation (En) was also computed in order to validate the method used. En values obtained ranged between 0.07 and 0.11 for mussel samples and between 0.26 and 0.72 for sediment samples. These values show that isotope dilution–GC–ICPMS methodology is valid for the determination of MBT, DBT and TBT from both types of matrices.

In this study the comparison of three different alkaline extraction techniques carried out with methanolic NaOH followed by phenylation derivatization for the determination of methylmercury in marine fish was performed. The investigated three methods differed in the technique how the extraction assisting energy was introduced to the sample. Namely, closed vessel ultrasonic bath, an open vessel technique using an ultrasonic probe and the microwave assisted extraction procedures were characterized and optimized. Optimum values of 3 h at 75 °C for the ultrasonic bath, 25 min for the ultrasonic probe and 6 min at 60 W for the microwave method were obtained. All three methods were validated using the BCR-464 tuna fish certified reference material.

Grapevine leaf synthesizes a wide variety of bioactive secondary metabolites, including polyphenols, which are also key components in ensuring development and growth of the whole plant even under adverse environmental conditions. Our study evaluates the nonanthocyanin polyphenolic composition in grapevine leaves of three varieties of Goher conculta (Vitis vinifera L.) native to Hungary. A high performance liquid chromatography (HPLC) system including a diode array detector (DAD) coupled to a time-of-flight mass spectrometer (q-TOFMS) was successfully applied to profile intact glycoconjugate forms in samples. In-source fragmentation was utilized in order to provide structural information on the compounds. Using this method, the presence of 16 polyphenolic metabolites were confirmed, and eight of them were subjected to further quantification in sun acclimated and half shaded leaves. Intracellular microimaging detected accumulation of flavonols in cell nuclei, cell wall and chloroplasts. Our findings demonstrated that Goher conculta—a special grapevine taxon of our viticultural heritage with berry color variants—is a suitable model to study the interaction between genetic and environmental factors in determination of grapevine phenolic composition.

The aims of the present study were to clarify in vivo effects of three sour cherry cultivars characterized by different polyphenolic composition in hyperlipidemic animals in a short term experiment. The three different sour cherry cultivars were chosen based on their total in vitro antioxidant capacity, total polyphenolic, monomeric anthocyanin and flavonoid content. Male Wistar rats were divided randomly into eight groups: rats kept on normal diet (control) and normal diet supplied with sour cherry powder of one of the three cultivars; others were kept on fat-rich diet and fat-rich diet supplied with sour cherry powder prepared from one of the three cultivars. The treatment lasted 10 days. Lyophilized sour cherry administered in the diet decreased both total cholesterol and LDL cholesterol levels, and increased the HDL cholesterol concentration in sera of hyperlipidemic animals. Significant differences were found in the efficacy of different sour cherry cultivars in case of hyperlipidemia. Sour cherries characterized by higher polyphenol content seem to have a more pronounced effect on serum cholesterol levels. Our results suggest that besides anthocyanins, colourless polyphenols also have lipid lowering effect.

Excessive post-prandial glucose concentration is associated with an increased risk of developing type 2 diabetes. Polyphenols represent a biologically active class of molecules which have been test...

Application of solid phase microextraction (SPME) to the extraction of Cr from sea-water after derivatization with the β-diketonate ligand 1,1,1-trifluoro-2,4-pentadione (trifluoroacetylacetone) is reported. The chelation reaction was conducted in a single (aqueous) phase medium. Both liquid phase and headspace extraction were studied by employing a 100 µm PDMS (polydimethylsiloxane) coated SPME fiber. Capillary gas chromatography (GC) coupled to electron capture detection (ECD), electron impact-mass spectrometry (EI-MS) and inductively coupled plasma-mass spectrometry (ICP-MS) were compared for the quantitation of Cr. Except for the ECD, isotope dilution calibration was applied, using 53Cr enriched spikes. Detection limits between 0.011–0.015 ng ml−1 (as Cr) were achieved with all three systems. Accuracy was assessed using CASS-4 Nearshore Seawater Certified Reference Material (CRM) from the National Research Council of Canada (NRCC). The developed method provided accurate results with EI-MS and ICP-MS detection, while significant bias was experienced with the ECD.

A simultaneous multi-elemental measurement of As, Bi, Ge, Sb, Se, and Sn was performed in this study by flow injection–hydride generation–inductively coupled plasma–time-of-flight mass spectrometry (FI–HG–ICP–TOFMS). An off-line pre-reduction treatment using a solution of 5% (m/v) KI and ascorbic acid for 15 min at 80 °C is described by presenting its advantages and disadvantages and compared with the results achieved without pre-reduction. Using optimised conditions the following figures of merit were achieved: limits of detection in the 0.08–0.54 ng ml −1 range and relative standard deviations (RSD) of 1.7–6.7%, respectively. Applying the presented method, two certified reference materials (NIST 1643d freshwater and PACS-2 marine sediment) were analysed to demonstrate the suitability of the method for analysis of real samples. Results obtained from treated samples showed good agreement with certified values while the untreated ones considerably departed from them.

In fields such as food and nutrition science or plant physiology, interest in untargeted profiling of flavonoids continues to expand. The group of flavonoids encompasses several thousands of chemically distinguishable compounds, among which are a number of isobaric compounds with the same elemental composition. Thus, the mass spectrometric identification of these compounds is challenging, especially when reference standards are not available to support their identification. Many different types of isomers of flavonoid glycoconjugates are known, i.e. compounds that differ in their glycosylation position, glycan sequence or type of interglycosidic linkage. This work focuses on the mass spectrometric identification of flavonoid glycoconjugate isomers possessing the same glycan mass and differing only in their aglycone core. A non-targeted HPLC-ESI-MS/MS profiling method using a triple quadrupole MS is presented herein, which utilizes in-source fragmentation and a pseudo-MS(3) approach for the selective analysis of flavonoid glycoconjugates with isomeric/isobaric aglycones. A selective MRM-based identification of the in-source formed isobaric aglycone fragments was established. Additionally, utilizing the precursor scanning capability of the employed triple quadrupole instrument, the developed method enabled the determination of the molecular weight of the studied intact flavonoid glycoconjugate. The versatility of the method was proven with various types of flavonoid aglycones, i.e. anthocyanins, flavonols, flavones, flavanones and isoflavones, along with their representative glycoconjugates. The developed method was also successfully applied to a commercially available sour cherry sample, in which 16 different glycoconjugates of pelargonidin, genistein, cyanidin, kaempferol and quercetin could be tentatively identified, including a number of compounds containing isomeric/isobaric aglycones.

The fruit quality parameters and antioxidant capacity (ferric reducing antioxidant power, FRAP) and total phenolic content (TPC) were determined in 27 apricot cultivars and hybrids of diverse origins. Twenty one- to 35-fold variations were measured among FRAP and TPC values. Besides genotype, harvest year also contributed significantly (P≤0.05) to the variations of TPC presumably due to the climatic differences between years. A subset of genotypes (15) was also analyzed for their antiradical activities (2,2'-diphenyl-1-picrylhydrazyl, DPPH; total radical-scavenging activity, TRSA; water-soluble antioxidant capacity, ACW; and lipid-soluble antioxidant capacity, ACL), and vitamin C contents as well as color indices (CIE H°, L*, and chroma). The hybrid "Preventa" had outstanding FRAP (>10.4 mmol ascorbic acid/L), DPPH (74.45%), TRSA (0.002%), ACW (33587.5 nmol AA/L) and ACL (78.65 nmol Trolox/L), TPC (>2890.0 mg gallic acid/L), and vitamin C (16.17 mg/100 g FW) levels and an average carotenoid content estimated from the hue angle (66.99°). Most antioxidant and antiradical activities correlated significantly except for TRSA; the closest correlation was observed between FRAP and ACW (r=0.952). Only TRSA showed significant correlations with color indices, H° and chroma, suggesting TRSA measures at least a fraction of the antioxidant capacity attributable to apricot carotenoids.