Identification of phenolic constituents of Cytisus multiflorus

The phenolic composition of the ethanolic extract obtained from the flowers of the medicinal plant Cytisus multiflorus has been elucidated by high performance liquid chromatography, electrospray mass spectrometry and nuclear magnetic resonance analysis. The extract was mainly composed of flavones, including the common chrysin, orientin, luteolin-5-O-glucoside, luteolin-7-O-glucoside, apigenin and apigenin-7-O-glucoside, which appeared as minor components. The major flavone in the extract was chrysin-7-O-β-d-glucopyranoside, and it also contained moderate amounts of a dihydroxyflavone isomer of chrysin, as well as of 2″-O-pentosyl-6-C-hexosyl-luteolin, 2″-O-pentosyl-8-C-hexosyl-luteolin and 6″-O-(3-hydroxy-3-methylglutaroyl)-2″-O-pentosyl-C-hexosyl-apigenin, which are not commonly found in the Fabaceae family. Other novel phenolic compounds found in the ethanolic extract of C. multiflorus comprised the flavones 2″-O-pentosyl-6-C-hexosyl-apigenin, 2″-O-pentosyl-8-C-hexosyl-apigenin and 6″-O-(3-hydroxy-3-methylglutaroyl)-2″-O-pentosyl-C-hexosyl-luteolin. The assessment of the biological activities of the main compounds of this extract are now keen, in order to determine their relevance in the beneficial properties of the plant.     

Identification of isomers of resveratrol dimer and their analogues from wine grapes by HPLC/MS and HPLC/DAD-UV

A facile method based on HPLC/(−)ESI-MSⁿ is established for the analysis of seven isomers of resveratrol dimers and three of their analogues in Xinjiang wine grapes. The structures of these compounds are positively or tentatively determined. Among them, three are tentatively identified as new compounds. MSⁿ experiments on the [M−H]⁻ ions provide abundant structural information, especially regarding the relative abundance of the key product ions, m/z 333 and 369 (385 in compound 3), which can be utilised to distinguish whether or not the compound identified contains the scaffold of the isomer of a resveratrol dimer. The relative abundance of key product ions remains unchanged as collision energy varies from 0.60 to 0.95 V. All the trans-, and cis-isomers could be identified by HPLC/DAD-UV spectra. The UV spectra of compounds 2 and 9 tentatively show cis and trans- configurations, respectively.     

An improved LC-MS/MS method for the quantitation of acrylamide in processed foods

An improved liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the determination of acrylamide in processed foods. The homogenized samples, spiked with ¹³C₃-acrylamide as an internal standard, were extracted with water and centrifuged. D₅-3-chloropropanediol as a recovery standard was added to 1-ml aliquots, and the sample was purified with a C₁₈-cartridge column. The extract was directly analyzed using LC-MS/MS without derivatization. The ion transitions of 72–55 m/z (acrylamide), 75–58 m/z (¹³C₃-acrylamide), and 116–98 m/z (d₅-3-chloropropanediol) were found to be the most reliable for the identification and quantification of acrylamide in multiple reaction monitoring. The limit of quantification for acrylamide, defined as a signal-to-noise ratio of 10:1, was 2 μg/kg. The use of d₅-3-chloropropanediol minimized the effects of variation in the sample matrixes and increased the quality of analysis. This method could be applied to the quantification of acrylamide in processed foods.     

Carotenoids and triacylglycerols interactions during thermal oxidation of refined olive oil

Carotenoids (β-carotene and astaxanthin) were added to refined olive oils and oxidised in a Rancimat at 110 °C for 1–14 h. Triacylglycerols (TAGs) were found to oxidise much faster in the presence of β-carotene and slower in the presence of astaxanthin, while β-carotene degradation was much faster than that of astaxanthin. Astaxanthin was found to protect the TAGs of olive oils for up to 10 h of thermal treatment. Both carotenoids were found to significantly increase peroxide values at higher exposure time. A total of 11 TAGs oxidised species were identified, including a new class of oxidised species (hydroxy epidioxides), reported here for the first time. The effect of carotenoids on the formation of oxidised TAGs illustrates that the pro-oxidant action of β-carotene was much stronger than that of astaxanthin. We suggest that astaxanthin could be used as an alternative to β-carotene in edible oils fortification and as a food colourant.     

Direct nanoHPLC-ESI-QTOF MS/MS analysis of tryptic caseinophosphopeptides

Caseinophosphopeptides (CPPs) were generated following tryptic hydrolysis of sodium caseinate. Hydrolysate peptides were separated and identified using nano-HPLC ESI-QTOF MS/MS. Sequence coverage in the 3 h hydrolysate was 79.4%, 55.6%, 80.9% and 68.1% for α_s1-, α_s2-, β- and κ-casein (CN), respectively. Variable levels of serine phosphorylation in β-CN f1–25 were observed in the 3 h hydrolysate. Analysis of β-CN f1–25 4P demonstrated that this peptide was stable during the course of hydrolysis. The effect of heat treatment (75 °C, 45 min) at pH 6.0, 7.0 and 8.0 on the peptide profile of the 3 h hydrolysate was studied. Compared to pH 6.0 and 8.0, least modification in phosphopeptide profiles was observed for the hydrolysate sample heated at pH 7.0. Different dephosphorylation and oxidation patterns were also observed following heat treatment at the three pH values. These results demonstrate that heat treatment, in addition to pH, has a major effect on both the phosphorylated and non-phosphorylated peptide profiles of CN hydrolysates.     

Tocols of selected spring wheat (Triticum aestivum L.), einkorn wheat (Triticum monococcum L.) and wild emmer (Triticum dicoccum Schuebl [Schrank]) varieties

Wheat contributes significantly worldwide to antioxidant income with beneficial healthy effects. Two varieties of einkorn wheat (Triticum monococcum L. – Escana and Schwedisches Einkorn), two varieties of wild emmer [Triticum dicoccum Schuebl (Schrank)] (Rudico and Kahler Emmer) and three varieties of spring wheat (Triticum aestivum L. – Granny, Kärtner Früher and SW Kadrilj) were evaluated for tocols – tocopherols and tocotrienols by high performance liquid chromatography – electrospray ionisation tandem mass spectrometry (HPLC–ESI/MS/MS). Investigated emmer and einkorn wheat varieties have shown minor lower total tocols content in comparison with spring varieties, however significant typical levels of tocopherols and tocotrienols were found in these groups. In einkorn wheat varieties the major abundance of total tocols is represented by tocotrienols (Schwedisches Einkorn 65.0%, Escana 54.9%), in emmer wheat varieties the abundance of tocotrienols and tocopherols were similar (Rudico 48.9% of tocotrienols, Kahler Emmer 47.6%), whereas in spring bread wheat higher tocopherols (SW Kadrilj 54.5%, Kärtner Früher 60.2%, Granny 55.0%) were recorded.     

Germination induces the glucosylation of the Fusarium mycotoxin deoxynivalenol in various grains

In food, the mycotoxin deoxynivalenol (DON) often occurs in conjunction with its 3-β-d-glucopyranoside (D3G). The transformation of DON to D3G through glucosylation is catalysed by plant enzymes, however, the exact circumstances are not well understood. In order to investigate the role of enzymatic glucosylation in germinating grains, DON treated kernels were steeped and germinated under laboratory conditions. Furthermore, the effect of malting on the DON content of the contaminated barley was investigated. In all cases, DON and its derivatives were quantified by HPLC-MS/MS before, during and after the experiments. Amongst the six tested cereals; wheat, rye, barley, spelt, and millet transformed DON to D3G during germination whilst the oats were inactive. For wheat, barley, and spelt the initial DON content was reduced by 50%, with the loss being almost entirely accounted for by D3G formation. As D3G might be cleaved during digestion, the elevated D3G concentration may obscure the toxicologically relevant DON content in processed food and beer. The germination process has a major influence on the “masking” of DON, leading to high quantities of D3G that may be missed in common mycotoxin analyses.     

Evidence for galloylated type-A procyanidins in grape seeds

In this work, procyanidins were extracted with methanol from the seeds of white and red grape varieties, and fractionated using graded methanol/chloroform precipitation in order to obtain the oligomers of lower molecular weight. These were analyzed by electrospray ionization-mass spectrometry (ESI-MS and ESI-MS/MS) in the positive mode. Protonated molecules, [M+H]⁺, of procyanidin species of nongalloylated and monogalloylated type-A and type-B oligomers, with degree of polymerisation 2–5, and digalloylated oligomers, with degree of polymerisation 2–3, were observed in the ESI-MS spectra. Type-A procyanidin abundance accounted for 60–80% of the abundance of the corresponding type-B species. Independent of the interflavanic linkage, the abundance of monogalloylated dimers accounted for 20% of the abundance of the corresponding nongalloylated ones. For the higher degrees of polymerisation, the abundance of galloylated oligomers was shown to reach up to 60% of the abundance of the corresponding nongalloylated oligomers. Thiolytic analyses showed that the type-A interflavanic linkages were present in the terminal units whereas the type-B interflavanic linkages were present as extension units. Although many reports are already available regarding the analyses of procyanidin polymers and grape seed procyanidins, this is the first report for the occurrence of type-A galloylated procyanidins. The similarity of the relative abundances and structural features observed in the samples of both white and red grape varieties, shows that these are characteristic of grape seeds.     

Naturally-occurring folates in foods: Method development and analysis using liquid chromatography–tandem mass spectrometry (LC–MS/MS)

An accurate method for detecting and quantifying both synthetic (folic acid) and naturally-occurring folates in foods is described. A system capable of analysing the five most commonly occurring folates (pteroylglutamic acid, 5-methyltetrahydrofolate, tetrahydrofolate, 10-formylfolate and 5-formyltetrahydrofolate) in 20 min using liquid chromatography–tandem mass spectrometry (LC–MS/MS) was developed. Quantification of folates was performed using ¹³C labelled internal standards. This paper outlines the development of a comparatively fast LC–MS/MS method, method validation using commercially available folate standards and establishment of the method’s suitability for quantification using selected reaction monitoring (SRM) mass spectrometry. The application of the system was verified by analysing several certified reference materials and comparing results with certified values as determined by microbiological assay. LC–MS/MS promises to be an ideal tool for the quantitative analysis of folates in food.     

Isolation and evaluation of the antioxidant activity of phenolic constituents of the Garcinia brasiliensis epicarp

A new glycosylated biflavonone, morelloflavone-4′″-O-β-d-glycosyl, and the known compounds 1,3,6,7-tetrahydroxyxanthone, morelloflavone (fukugetin) and morelloflavone-7″-O-β-d-glycosyl (fukugeside) were isolated from the epicarp of Garcinia brasiliensis collected in Brazil. The structures of these compounds were established using ¹H and ¹³C NMR, COSY, gHMQC and gHMBC spectroscopy. The compounds exhibited antioxidant activity. The greatest potency was displayed by morelloflavone (2), with IC₅₀ = 49.5 mM against DPPH and absorbance of 0.583 at 400 μg/mL for the reduction of Fe³⁺. The weakest potency was displayed by 1,3,6,7-tetrahydroxyxanthone (1), with IC₅₀ = 148 mM against DPPH and absorbance of 0.194 at 400 μg/mL for the reduction of Fe³⁺.     

Comparison of methods for the microbiological identification and typing of Cronobacter species in infant formula

Cronobacter species represent an emerging opportunistic foodborne pathogen associated with meningitis and necrotizing enterocolitis in infants. Current evidence indicates that powdered infant formula (PIF) is the main source of Cronobacter contamination. A total of 75 strains of Cronobacter spp. from different geographic regions, as well as from PIF processing environments, were identified and typed with different methods, including biochemical profiling by the API 20E system (bioMérieux, Marcy l’Etoile, France), protein profiling by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and genotypic profiling by ribotype. Analysis by MALDI-TOF MS and biochemical identification was more accurate compared with ribotype analysis. However, MALDI-TOF MS typing and ribotype analysis showed more discriminatory ability compared with biochemical phenotyping. In conclusion, MALDI-TOF MS is a rapid and reliable tool to identify Cronobacter spp. in PIF and has the potential to trace dissemination of Chronobacter along the production chain.     

Fumonisin contamination and fumonisin producing black Aspergilli in dried vine fruits of different origin

Aspergillus niger isolates are able to produce fumonisins in high quantities on agar media with a low water activity. Several agricultural products fit this criterion, including dried vine fruits, dates and figs. Data on the occurrence and role of this species in fumonisin contamination of agricultural products with high sugar content are needed to clarify the importance of A. niger in human health. The mycobiota and fumonisin contamination of various dried vine fruit samples collected from different countries were examined to clarify the role of black Aspergilli in fumonisin contamination of such products. All except two of the examined samples were contaminated with black Aspergilli. Species assignment of the isolates was carried out using sequence analysis of part of the calmodulin gene. The range of fumonisin isomers present in the raisin samples, and produced by A. niger isolates collected from dried vine fruits was also examined using reversed-phase high-performance liquid chromatography/electrospray ionization-ion trap mass spectrometry (RP-HPLC/ESI-ITMS). Among the 30 A. niger/A. awamori isolates identified, 20 were found to be able to produce fumonisins (average contamination: 5.16 mg/kg; range: 0.017-19.6 mg/kg). The average fumonisin content of the 7 dried vine fruit samples which were found to be contaminated by potential fumonisin producing black Aspergilli was 7.22 mg/kg (range: 4.55-35.49 mg/kg). The isolates produced several fumonisin isomers also present in the dried vine fruit samples, including fumonisins B_1-4, 3-epi-FB₃, 3-epi-FB₄, iso-FB₁, and two iso-FB_2,3 forms. Fumonisin B₁ was detected for the first time in A. niger cultures. Most of these isomers have previously only been identified in Fusarium species. Our data indicate that A. niger and A. awamori are responsible for fumonisin contamination of dried vine fruits worldwide. The observed levels of contamination are alarming and pose a new threat for food safety.     

Characterisation of volatile and non-volatile metabolites in etiolated leaves of tea (Camellia sinensis) plants in the dark

Aroma is an essential factor affecting the quality of tea (Camellia sinensis) products. While changes of volatile compounds during tea manufacturing have been intensively studied, the effect of environmental factors on volatile contents of fresh tea leaves has received less attention. We found that C. sinensis var. Yabukita kept in darkness by shading treatment for 3 weeks developed etiolated leaves with significantly increased levels of volatiles, especially volatile phenylpropanoids/benzenoids (VPBs). Upstream metabolites of VPBs, in particular shikimic acid, prephenic acid, and phenylpyruvic acid, showed lower levels in dark treated than in control leaves, whereas the contents of most amino acids including l-phenylalanine, a key precursor of VPBs, were significantly enhanced. In addition, analysis by ultra performance liquid chromatography-time of flight mass spectrometry, capillary electrophoresis–time of flight mass spectrometry, high performance liquid chromatography, and gas chromatography–mass spectrometry indicated that volatile and non-volatile metabolite profiles differed significantly between dark treated and untreated leaves.     

Quinic acid esters and sesquiterpenes from Sonchus arvensis

A phytochemical study on the whole plant of Sonchus arvensis and its antioxidant activity has been carried out. Three quinic acid derivatives (1–3), the rarely naturally occurring (p-hydroxyphenylacetyl) quinic acids, and two eudesmanolides (4 and 5) were newly found. Four known eudesmanolides (6–9) were isolated from the plant for the first time. Their structures were characterized by HRESIMS, IR, UV, 1D NMR, and 2D NMR. 1,1-Diphenyl-2-picrylhydrazyl (DPPH·)-scavenging activity was evaluated for each of the above 9 compounds (1–9) in comparison to standard antioxidants (caffeic acid and ascorbic acid). However, none proved to have a positive activity. The absence of antioxidant activity could be caused by the absence of ortho or para-diphenolic groups in all detected compounds, that are responsible of the activity against free radicals by an electron transfer reaction.     

Phenolic acid contents of kale (Brassica oleraceae L. var. acephala DC.) extracts and their antioxidant and antibacterial activities

Nine phenolic acids were identified and quantified by HPLC–MS in leaves and 10 in seeds of kale (black cabbage). The free, ester (methanol-soluble), glycoside and ester-bound (methanol-insoluble) phenolic acid contents of the leaves were 487, 532, 4989 and 6402 ng/g fresh weight, respectively. Ferulic and caffeic acids (total contents; 4269 and 4887 ng/g, respectively) were the most abundant. The seed contents of these fractions were 1993, 1477, 1231 and 4909 ng/g dry weight (DW), respectively, and sinapic acid was the most abundant (5037 ng/g DW). The fractions’ total phenolic contents, determined colorimetrically, were highly correlated with their DPPH scavenging capacity, and in antimicrobial activity assays, with nine test organisms representing a wide array of taxa, all of the fractions were effective against Staphylococcus aureus, Enterococcus faecalis, Bacillus subtilis and (most strongly) Moraxella catarrhalis. Antimicrobial and antioxidant activities of kale phenolics in free and conjugated forms are discussed.     

Anthocyanins composition and antioxidant activity of two major wild Nitraria tangutorun Bobr. variations from Qinghai-Tibet Plateau

Nitraria tangutorun Bobr., a unique kind of fruit, widely spreads in the Qinghai-Tibet Plateau. In the present study, nine anthocyanins were identified in two variations (purple fruit and red fruit) of N. tangutorun by HPLC/DAD-ESI/MS. Cyanidin-3-O-(trans-p-coumaroyl)-diglucoside (215.76 ± 22.91 mg of Mv3G5G equivalent per 100 g of fresh weight) and pelargonidin-3-O-(p-coumaroyl)-diglucoside (5.13 ± 0.35 mg of Mv3G5G equivalent per 100 g of fresh weight) were the main anthocyanins in the purple and red fruits respectively. In addition, most of the anthocyanins were acylated by coumaric acid, and the rare anthocyanins that naturally presented a coumaric acid in both cis and trans configurations have been detected. Furthermore, the extract of the two variations showed significantly different antioxidant activity (p < 0.01) according to DPPH, ABTS and FRAP assay. Purple fruit possessed higher antioxidant activity than red fruit. There were significant correlations between antioxidant activity and both the total polyphenol content and anthocyanins content. This work is valuable for elucidation of anthocyanins composition in N. tangutorun and for further utilization as a functional food and medicine material.     

Diet and plasma evaluation of the main isomers of conjugated linoleic acid and trans-fatty acids in a population sample from Mediterranean north-east Spain

The aim of the present study was to describe the dietary pattern of a representative sample of 516 adult participants (203 men and 313 women) from Catalonia, a Spanish Mediterranean region, to assess their current dietary and plasma levels of trans C18:1, the major trans-fatty acid (TFA), and cis-9, trans-11 CLA, and trans-10, cis-12 CLA, the two major conjugated linoleic acid (CLA) isomers, and to evaluate their correlation with several cardiovascular disease risk factors. The population was a random sample derived from the Catalan Nutrition Survey. Plasma levels of the CLA isomers were determined in a subsample of 100 volunteers. The Catalan diet seemed to maintain some traits of the ‘traditional’ Mediterranean diet, although other components were lost. The dietary intakes of saturated fatty acids (SFA), TFA, cis-9, trans-11 CLA, and trans-10, cis-12 CLA were 12.3%, 0.84% (2.0 g/d), 0.030% (71.5 mg/d), and 0.0015% (3.4 mg/d) of the energy intake, respectively. Trans C18:1 accounted for 0.19% of the total plasma fatty acids, while the sum of cis-9, trans-11and trans-10, cis-12 CLA isomers represented about 0.09% of the plasma fatty acids. Trans C18:1 isomers correlated significantly with the intake of French fries and pastries, while cis-9, trans-11 CLA significantly correlated with the intake of dairy products and ruminant meat. None of the cardiovascular disease risk factors were found to be associated with the plasma levels of TFA or CLA. The results of this study suggest that monounsaturated fatty acids (MUFA) are the main dietary fat source in the Catalan population, due to their regular olive oil consumption. Moreover, plasma levels of the main TFA and CLA suggest that the Catalan diet is not at present strongly influenced by the occidental dietary patterns. However, a reduction of the intake of SFA in the Catalan population should be recommended.     

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.     

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.     

Mass spectrometry based proteomics as foodomics tool in research and assurance of food quality and safety

BackgroundAs a comprehensive discipline that studies food and nutrition, foodomics requires reliable qualitative and quantitative information about the food proteome component in order to extract new integrative information from the complex multivariable space of omics. This new information is necessary to achieve a higher level of understanding of processes in food science and technology, consequently new functions of food and improved markers of food quality and safety and completely transform concept of food safety.Scope and approachWe are making an effort to present mass spectrometry (MS) based proteomic approaches that are being utilized in different proteomic studies, not necessarily in the field of foodomics, which are important and have the potential to advance this field. Current analytical capabilities of MS-based proteomics together with sample preparation procedures and quantification strategies, and recent technical developments were presented.Key findings and conclusionsMS-based proteomics enables the analysis of different aspects of proteins and provides a variety of approaches for reliable quantification of individual proteins and/or food proteome. This is a complex field and its successful implementation requires a dedicated analyst, a thorough design of sample preparation procedure, the selection of an MS technique and approach, an adequate type of mass spectrometer, a thorough data analysis and validation. Improvements in the technology of mass spectrometers are continuously expanding capabilities of MS-based proteomics.