Search

Introduction: Benzoxazinoids (BXs) are plant phytochemicals that have both defensive properties in plants and therapeutic effects in humans. The presence of BXs has been largely studied in the Poaceae family (monocots). To study the presence or absence of BXs in dicotyledons and monocotyledons outside the Poaceae family, parts of 24 plant species at several growth stages were selected for analysis, some of which were already known to contain BXs.Objectives: To devise a step-wise mass spectrometry-based approach for confirming the presence of BXs in plant samples, and to use the method to explore the status of BXs in selected plant species.Experimental: Plant samples were extracted using accelerated solvent extraction and analyzed using triple-quadrupole liquid chromatography-mass spectrometry.Results: The use of different columns, double mass transitions, and ion ratios proved to be a robust tool for confirming the presence of BXs in different plant species. By this method, the presence of BXs was confirmed in three of the 24 species. Double-hexose forms of BXs, which have not been reported before in dicotyledons, were confirmed to be present in the dicotyledon plants Acanthus mollis and Lamium galeobdolon, and the presence of BXs in the seeds of Consolida orientalis is reported for the first time here. High concentrations of BXs were found in the aerial parts of Acanthus mollis and Lamium galeobdolon, at 20 and 32 μmol/g plant dry weight, respectively.Conclusions: The stepwise approach described in this work confirmed the presence of BXs in new samples.

Phytochemicals or phenolic compounds are important natural bioactive molecules that plants accumulate in response to environmental conditions and may exert benef icial effects on health by protecting humans against many diseases. The aim of this work was to analyze the influence of biotic and abiotic stress on the accumulation of flavonoids and phenolic acids on leaves of two cultivars of Amaranthus hypochondriacus, which are differentiated by the colour of their leaves (red or green). Phenolic compounds were extracted using the accelerated solvent extraction (ASE) method and their identif ication was carried out by LC-MS analysis. Rutin was the main flavonoid in amaranth leaves; the highest concentrations were found in green leaves when plants were subjected to stress (9715 μg g-1). Phenolic acids were minor compounds; ferulic acid was only present in red leaves (0.5 μg g-1) and p-coumaric acid only in green leaves (0.7 μg g-1). Our results indicate that leaves from A. hypochondriacus, the main species that produce edible seeds, are a good source of phytochemical compounds and their accumulation could be driven by the growing conditions.

Honey bees are important pollinators and are subject to numerous stressors, such as changing floral resources, parasites, and agrochemical exposure. Pesticide exposure has been linked to the decline in the global honey bee population. We have limited knowledge of the metabolic pathways and synergistic effects of xenobiotics in bees. Quercetin is one of the most abundant phytochemicals in plants and is therefore abundant in the honey bee diet. Quercetin can upregulate the detoxification system in honey bees; however, it is still unknown to what extent quercetin ingestion can reduce the content of absorbed pesticides. In this study, we investigated the effect of dietary quercetin on the contents of three pesticides in honey bees: imidacloprid (insecticide), tebuconazole (fungicide), and tau-fluvalinate (insecticide and acaricide). Bees were divided into two main groups and fed either quercetin-sucrose paste or only sucrose for 72 h. Thereafter, they were orally exposed to ∼10 ng/bee imidacloprid or contact-exposed to ∼ 0.9 μg/bee tau-fluvalinate or ∼ 5.2 μg/bee tebuconazole. After 1 h of oral exposure or 24 h of contact exposure, the bees were anaesthetised with CO2, sacrificed by freezing, and extracted with a validated QuEChERS method. Subsequently, the concentrations of the three pesticides and quercetin in the bees were determined with a triple quadrupole tandem mass spectrometer coupled to an HPLC system. No significant effect on the concentration of tebuconazole or tau-fluvalinate was observed in bees fed quercetin. Intake of quercetin led to a reduction in the concentration of imidacloprid in honey bees. Quercetin-rich plants may be exploited in future beekeeping.

The honey bee pollen/nectar diet is rich in bioactive phytochemicals and recent studies have demonstrated the potential of phytochemicals to influence honey bee disease resistance. To unravel the role of dietary phytochemicals in honey bee health it is essential to understand phytochemical uptake, bioavailability, and metabolism but presently limited knowledge exists. With this study we aim to build a knowledge foundation. For 5 days, we continuously fed honey bees on eight individual phytochemicals and measured the concentrations in whole and dissected bees by HPLC-MS/MS. Ample phytochemical metabolization was observed, and only 6-30% of the consumed quantities were recovered. Clear differences in metabolization rates were evident, with atropine, aucubin, and triptolide displaying significantly slower metabolism. Phytochemical gut uptake was also demonstrated, and oral bioavailability was 4-31%, with the highest percentages observed for amygdalin, triptolide, and aucubin. We conclude that differences in the chemical properties and structure impact phytochemical uptake and metabolism.

Honey bees regularly ingest quercetin, one of the most abundant phytochemicals in plants. Past research suggests that honey and pollen feeding upregulates the bees’ detoxification system and that dietary quercetin reduces the toxicity of imidacloprid and tau-fluvalinate [1,2].The aim of this study was to investigate the effect of dietary quercetin on the concentration of three pesticides present in honey bees. Honey bees (n = 600) were divided in five groups and fed either quercetin-sucrose paste or only sucrose for 72 h before being exposed to the neonicotinoid imidacloprid (oral exposure), the fungicide tebuconazole or the acaricide tau-fluvalinate (contact exposure). Bees were anesthetized with CO2, frozen, and extracted with a validated QuEChERS method. Finally, the concentrations of all three above-mentioned pesticides were determined by LC-MSMS. Quantification results of the three pesticides in honey bee samples [Fig. 1] displayed that there was a significant difference in concentration of imidacloprid (P = 2 ͯ 10-5) and tau-fluvalinate (P = 3 ͯ 10-2) between the bees that were fed quercetin-sucrose paste and bees that were fed only sucrose. However, there was no significant difference in the level of tebuconazole in the group of bees that were fed quercetin-sucrose paste and bees that were fed only sucrose (P = 4.2 ͯ 10-1). The results of this study demonstrate that honey bee intake of quercetin leads to a reduction in the concentration of imidacloprid and tau-fluvalinate by up regulating the detoxification systems in bees. Hence, quercetin may be a promising bioactive compound in chemo-ecological aspects to counteract the hazard of pesticides. Honey bees regularly ingest quercetin, one of the most abundant phytochemicals in plants. Past research suggests that honey and pollen feeding upregulates the bees’ detoxification system and that dietary quercetin reduces the toxicity of imidacloprid and tau-fluvalinate [1,2].The aim of this study was to investigate the effect of dietary quercetin on the concentration of three pesticides present in honey bees. Honey bees (n = 600) were divided in five groups and fed either quercetin-sucrose paste or only sucrose for 72 h before being exposed to the neonicotinoid imidacloprid (oral exposure), the fungicide tebuconazole or the acaricide tau-fluvalinate (contact exposure). Bees were anesthetized with CO2, frozen, and extracted with a validated QuEChERS method. Finally, the concentrations of all three above-mentioned pesticides were determined by LC-MSMS. Quantification results of the three pesticides in honey bee samples [Fig. 1] displayed that there was a significant difference in concentration of imidacloprid (P = 2 ͯ 10-5) and tau-fluvalinate (P = 3 ͯ 10-2) between the bees that were fed quercetin-sucrose paste and bees that were fed only sucrose. However, there was no significant difference in the level of tebuconazole in the group of bees that were fed quercetin-sucrose paste and bees that were fed only sucrose (P = 4.2 ͯ 10-1). The results of this study demonstrate that honey bee intake of quercetin leads to a reduction in the concentration of imidacloprid and tau-fluvalinate by up regulating the detoxification systems in bees. Hence, quercetin may be a promising bioactive compound in chemo-ecological aspects to counteract the hazard of pesticides.

The presence of bioactive benzoxazinoids (BXs) in mature whole grain and experimental bread products was recently shown. The aim of this study was to reveal the level of BXs in a broad range of commercial cereal products and discuss these findings in relation to ingredients and preparation processes. The concentrations of 11 BX compounds were determined by liquid chromatography–triple quadrupole mass spectrometry (LC–MS/MS) in 25 supermarket-available cereal food products. The total BX content varied from below the limit of detection to >500 μg/g dry matter (DM) with some degree of variation among different lots. The products W (Rugsandwich), H (Gulerodsrugbrod), L (Morke Multikerneboller) and AA (Solsikkeklapper), which were all rich in rye-based ingredients, had a total BX concentration >300 μg/g DM. Double hexose derivatives of 2-hydroxy-1,4-benzoxazin-3-one (HBOA), 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA) and 2-β- d -glucopyranosyloxy-4-hydroxy-1,4-benzoxazin-3-one (DIBOA-glc) were the major BXs in most products. Further studies into the health effects of BXs should be conducted, and future exploitation of these compounds seems possible.

OBJECTIVES: Prostate cancer is a common cancer diagnosis among men, but the role of diet and food components remains controversial. Benzoxazinoids (BXs) is a potent group of bioactive compounds found in rye foods that seem to have the potential to inhibit prostate cancer growth in vitro but studies in humans are lacking. We therefore aimed to investigate the concentrations of different BXs and their metabolites after whole grain rye consumption in comparison to refined wheat and to correlate these compounds to PSA levels in men with indolent prostate cancer. METHODS: In a randomized controlled crossover study, men with prostate cancer (n = 24) consumed 485 gram of whole grain/bran rye and refined wheat foods for six weeks, with two weeks of washout in-between. The diets were isocaloric and contained the same amount of fiber by addition of cellulose in the refined wheat intervention. BXs in plasma samples were analyzed with HPLC- QTRAP/MS-ESI. RESULTS: Seventeen men finished the trial and were included in data analysis. For most BXs, the concentrations were significantly higher after the whole grain rye compared to the refined wheat intervention. The BXs or BXs metabolites HBOA-glc, HHPAA, HBOA-glcA, HPAA-glcA (∼19.4–0.4 nmol/L in plasma after interventions) were significantly inversely correlated to PSA level. CONCLUSIONS: In men with indolent prostate cancer, most BXs were significantly higher after an intervention with whole grain rye compared to refined wheat. The inverse correlation between four BXs or BXs metabolites and PSA may indicate on an inhibitory effect on prostate cancer cell growth. This could open for new preventive food-based strategies, but results would need to be replicated. FUNDING SOURCES: Independent Research Fund Denmark - Technology and Production Formas Swedish Research Council.

Benzoxanoids (BXs) were identified for the first time in young cereal plants in 1957 and in medicinal plants in 1969. In 2008 our lab discovered that BXs are present in mature cereal grains and in food/feed based on cereal whole grains [1]. We showed that BXs are taken up in pigs, rats and humans [2]–[4], and that they appear in prostate tissue of humans consuming rye bread [5]. The aim of this study was to develop a method for identification and quantification of human phase 1 and phase 2 metabolites in plasma. The method included an initial protein precipitation step of the plasma with the purpose of reducing the matrix interference. The chromatographic analysis of BXs and their metabolites in the remaining extract was performed in LC-MS/MS. Pure standards were unavailable for most of the phase 2 metabolites. Phase 2 metabolites are present in much higher amounts in urine than in plasma. Thus urine metabolites of BXs for which structures could be assigned using data dependent acquisition were isolated and used for verification of MS/MS signals in plasma extracts. The isolation of approx 10 µg of each compound was done with consecutive HLB and mixed mode SPE’s (Oasis, Waters) exploiting the variety of polarity and acidity of the metabolites. Glucuronide and sulphate metabolites of paracetamol were used as standards for semi-quantification due to their structural similarity and commercial availability. Benzoxanoids (BXs) were identified for the first time in young cereal plants in 1957 and in medicinal plants in 1969. In 2008 our lab discovered that BXs are present in mature cereal grains and in food/feed based on cereal whole grains [1]. We showed that BXs are taken up in pigs, rats and humans [2]–[4], and that they appear in prostate tissue of humans consuming rye bread [5]. The aim of this study was to develop a method for identification and quantification of human phase 1 and phase 2 metabolites in plasma. The method included an initial protein precipitation step of the plasma with the purpose of reducing the matrix interference. The chromatographic analysis of BXs and their metabolites in the remaining extract was performed in LC-MS/MS. Pure standards were unavailable for most of the phase 2 metabolites. Phase 2 metabolites are present in much higher amounts in urine than in plasma. Thus urine metabolites of BXs for which structures could be assigned using data dependent acquisition were isolated and used for verification of MS/MS signals in plasma extracts. The isolation of approx 10 µg of each compound was done with consecutive HLB and mixed mode SPE’s (Oasis, Waters) exploiting the variety of polarity and acidity of the metabolites. Glucuronide and sulphate metabolites of paracetamol were used as standards for semi-quantification due to their structural similarity and commercial availability.

Fusarium head blight (FHB) is a devastating disease of wheat (Triticum aestivum L.) caused by a mycotoxigenic fungus Fusarium graminearum resulting in significantly decreased yields and accumulation of toxic trichothecenes in grains. We tested 7 major secondary metabolites from wheat for their effect on trichothecene production in liquid cultures of F. graminearum producing trichothecene 15-acetyldeoxynivalenol (15-ADON). 2,4-Dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA) benzoxazinoid completely abolished toxin production without any apparent effect on fungal growth. DIMBOA strongly affected the expression of Tri6, encoding a major transcriptional regulator of several genes of the trichothecene biosynthesis pathway. DIMBOA also repressed expression of Tri5, encoding trichodiene synthase, the first enzyme in the trichothecene biosynthesis pathway. Thus, DIMBOA could play an important role against the accumulation of trichothecenes in wheat grain. Breeding or engineering of wheat with increased levels of benzoxazinoids could provide varieties with increased resistance against trichothecene contamination of grain and lower susceptibility to FHB.

Neonicotinoids are the most widely applied class of insecticides in cocoa farming in Ghana. Despite the intensive application of these insecticides, knowledge of their fate in the Ghanaian and sub-Saharan African environment remains low. This study examined the behavior of neonicotinoids in soils from cocoa plantations in Ghana by estimating their sorption and degradation using established kinetic models and isotherms. Studies of sorption were conducted using the batch equilibrium method on imidacloprid, thiamethoxam, clothianidin, acetamiprid and thiacloprid, while degradation of imidacloprid, thiamethoxam and their respective deuterated counterparts was studied using models proposed by the European forum for coordination of pesticide fate and their use (FOCUS). Analytes were extracted using the quick, easy, cheap, effective, rugged and safe (QuEChERS) procedure and quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Average recoveries were high (≥ 85%) for all analytes. The findings from the study suggest that neonicotinoid insecticides may be persistent in the soils studied based on estimated half-lives150 days. The study also revealed generally low-sorption coefficients for neonicotinoids in soils, largely influenced by soil organic carbon.

Benzoxazinoids (BXs) are a group of wholegrain phytochemicals with potential pharmacological properties; however, limited information exists on their absorption, metabolism, and excretion in humans. The aim of this study was to investigate the dose-dependent uptake and excretion of dietary BXs in a healthy population. Blood and urine were collected from 19 healthy participants from a crossover study after a washout, a LOW BX diet or HIGH BX diet, and analysed for 12 BXs and 4 phenoxazinone derivatives. We found that the plasma BX level peaked approximately 3 h after food intake, whereas BXs in urine were present even at 36 h after consuming a meal. No phenoxazinone derivatives could be detected in either plasma or urine. The dominant BX metabolite in both plasma and urine was 2-β-d-glucopyranosyloxy-1,4-benzoxazin-3-one (HBOA-Glc), even though 2-β-d-glucopyranosyloxy-4-hydroxy-1,4-benzoxazin-3-one (DIBOA-Glc) was the major component in the diet. The dietary BX treatment correlated well with the plasma and urine levels, illustrating strong dose-dependent BX absorption, which also had a rapid washout, especially from the plasma compartment.

SCOPE: To examine potentially immunomodulating effects of dietary benzoxazinoids (BXs), present in cereal grains.METHODS AND RESULTS: Nineteen healthy volunteers were randomly distributed into two groups, who received diets with high or low content of BXs for three weeks. After a week's wash-out, the groups switched diets. Peripheral blood mononuclear cells (PBMCs) were stimulated with Porphyromonas gingivalis, Escherichia coli lipopolysaccharide (LPS), or tetanus toxoid (TT). PBMCs from a healthy donor received the same stimuli in presence of serum from each participant receiving BXs. The production of monokines, T-cell cytokines and T-helper cell proliferation were assessed. A three-week diet with high BX content enhanced IL-1β responses against LPS and P. gingivalis, as well as TNF-α response against P. gingivalis, after 24 hours of stimulation. Moreover, IL-6 was found to be increased after 7 days of stimulation with LPS. No effect was observed on T-cell cytokines or proliferation. BX levels in serum after a single meal did not modify cytokine responses.CONCLUSIONS: High dietary intake of BXs enhances bacteria-induced production of pro-inflammatory monokines by PBMCs, but not T-cell responses; presumably due to intrinsic changes within PBMCs, built up over three weeks of BX-rich diet, rather than to immediate effects of BXs contained in serum. This article is protected by copyright. All rights reserved.

Lettuce is an important part of the diet in Europe. The permitted levels of pesticides in lettuce are strictly regulated and there is growing urge among food safety authorities to analyse pesticide metabolites as well. Azoxystrobin is one of pesticides that is frequently detected in lettuce. Although there are several analytical methods for the determination of azoxystrobin in lettuce, a sensitive method for the determination of its metabolites in lettuce is lacking. This study aimed at developing an extraction and LC-MS/MS method for the simultaneous determination of azoxystrobin, and its metabolites azoxystrobin free acid and 2-hydroxybenzonitrile in lettuce. Accelerated solvent extraction, QuEChERS extraction, and shaking extraction were compared using various solvents. The final method consisted of shaking freeze-dried sample in 0.1% formic acid in 80% aqueous acetonitrile. The selected method was validated by spiking each analyte at 125 ng/g and 500 ng/g. The method resulted in acceptable recovery for 2-hydroxybenzonitrile, azoxystrobin free acid, and azoxystrobin, with a RSD of

Benzoxazinoids are important phytochemicals found in wheat and rye that are associated with plant resistance against pathogens, and recent studies have emphasized the potential health-promoting role of these compounds i.e. anti-cancer, anti-allergy and anti-inflammation. Accordingly, an understanding of their distribution in seeds and the effect of different processing techniques on their transformation will be helpful in identifying the mechanisms of their production and distribution and will support the on-going efforts to utilize these compounds in health-promoting food products.The analysis of seed fractions obtained from the milling of wheat and rye showed significantly higher concentrations of these bioactive compounds in the germ than in the other fractions, i.e. the bran and endosperm. Di-hexoses of 2,4-dihydroxy-1, 4-benzoxazin-3-one (DIBOA-glc-hexose) and 2-hydroxy-1, 4-benzoxazin-3-one (HBOA-glc-hexose) were the predominant compounds found in the different wheat and rye seed fractions followed by DIBOA-glc and DIBOA.The soaking and boiling of three rye-based breakfast cereals resulted in considerable changes in the benzoxazinoid contents. The soaking of pearled rye promoted the conversion of DIBOA-glc-hexose into DIBOA-glc. When these cereals were boiled, the increase in the DIBOA-glc content was much lower than that observed for soaking. For rye flakes, the pattern of these benzoxazinoids was different from that in pearled rye seeds. A considerable amount of the benzoxazinoids was also leached into the water during soaking or boiling. This study contributes to the understanding of the underlying processes involved in the biochemical changes of benzoxazinoids and will be the basis for future studies on other food-processing techniques.

In line of the development of a food composition database for Bangladesh, 10 minor indigenous fruits were analysed for their nutrient composition comprising ascorbic acid, carotenoids and mineral values. Nutrient data obtained have been compared with published data reported in different literatures, book and United States Department of Agriculture-National Nutrient Database for Standard Reference. Ascorbic acid was highest in Wood apple and lowest in Roselle. Monkey jack contained the highest amount of carotenoids, zinc and copper. Content of calcium, magnesium and phosphorous were found highest in Antidesma velutinum. Potassium was the highest in Wood apple followed by in Moneky jack. It was noted that most of the minor fruits have much higher amount of ascorbic acid than the national fruit – Jack fruit ripe, the king fruit – Mango ripe of Bangladesh and exotic fruits – Apple and Grapes. The nutrient values of these minor fruits would make awareness among the people for their mass consumption for healthy life and to grow more minor fruit trees from extinction in order to maintain biodiversity.

This study was conducted to answer the question of whether clover can absorb asparagine in the presence and absence of inorganic nitrogen, as well as to determine the resulting concentration of post-uptake compounds closely involved in asparagine metabolism. Clover was grown at two asparagine concentrations (10 μM and 1 mM) supplied in both the absence and presence of ammonium nitrate. Using dual-labeled 13C15N-asparagine, the uptake rate was analyzed via bulk 15N and 13C excess and the detection of intact 13C15N-asparagine in white clover. The results from the two methods indicated greater utilization of 13C15N-asparagine in the 10 μM treatment than in the 1 mM treatment. The 13C15N-asparagine uptake rate was higher when 13C15N-asparagine was provided alone than when it was supplemented with inorganic nitrogen. Up to nine times lower uptake rates were obtained when intact 13C15N-asparagine was measured than when bulk 15N and 13C excess were analyzed. The labeled amino acids that are closely related to 13C15N-asparagine metabolism (aspartic acid, glutamic acid and glutamine) were detected in clover roots and shoots. Using two different methods, white clover’s potential to absorb intact asparagine, even in the presence of inorganic nitrogen, was confirmed. The dual-methodology approach employed in this study demonstrates how the post-uptake metabolism can affect quantification of amino acid uptake.

In a cultivation experiment 18 different Amaranthus genotypes were cultivated in parallel in Argentina, Mexico, Spain and two different locations in the Czech Republic. The ripe seeds were analysed for their content of 11 polyphenols and the variations among genotype, species and location were analysed by principal component analysis (PCA). The flavonoid, rutin, exhibited large variations with varying environmental conditions whereas the flavonoid, nicotiflorin, was affected less. Amaranthus hypochondriacus displayed the most stable content of polyphenols with a high end content of flavonoids. The variations between location/environmental condition were primarily described by the variations in the content of p-coumaric acid and protocatechuic acid in the seed samples.

Bread is consumed in large quantities all over the world, and rye bread is especially popular throughout the Nordic countries. Wholemeal bread is highly recommended as a basic ingredient in daily food, because wholemeal food products generally promote good health due to their vitamin, mineral and fibre content. The literature suggests that wholemeal products have other health-promoting effects even if the ingredients responsible have not been identified. Benzoxazinoids are a group of natural products that have not previously been reported in mature grains. Here, we report for the first time the identity and quantity of 10 compounds of the benzoxazinoid family in mature grains, hydrothermally processed grains of durum wheat ( Triticum durum , cv. Kamut), a commercial variety of rye ( Secale cereale cv. Picasso) and an old Nordic rye landrace ( S. cereale , Svedjerug), as well as in bread baked with flour milled from those grains. Concentrations of the 10 benzoxanoids were determined using LC–MS/MS and ranged from 0 to 348 nmol g −1 for conventional flour, to 772–1177 nmol g −1 in bread baked with flour from hydrothermally processed grains and to 3116–5570 nmol g −1 in flour from hydrothermally processed grains. Benzoxazinoids possess documented physiological effects, and research into the importance of these compounds in the daily diet is therefore needed. Ongoing studies in our lab on the uptake and transformation of benzoxazinoids in mammals will be reported in the near future.

The proximate composition, phenolic acid, and flavonoid characterization of two commercial and eight wild Opuntia spp., as well two processed products, were analyzed. Commercial varieties were obtained from a local market; wild varieties were collected from the Central and High Plateau zones of Mexico, and Opuntia tablets were purchased at a natural commodities store. Spines from fresh cladodes were removed and clean cladodes were freeze-dried and milled to obtain nopal powder. Differences in proximate composition were determined: wild nopal blanco collected from the High Plateau had the highest protein content, while tapon-II had the highest fiber content. Nopal tablets had low protein and carbohydrate content but had the highest ash content. The wild morado, tempranillo, blanco, and cristalino varieties had the highest total phenolic acid content, while the commercial varieties had the highest total flavonoid contents. Very low amounts of flavonoids were found in nopal tablets. LC-MS/MS was used to identify six phenolic acids (gallic, coumaric, 3,4-dihydroxybenzoic, 4-hydroxybenzoic, ferulic, and salicylic acid), and only tapon-II contained all six. The five flavonoids (iso-quercitrin, isorhamnetin-3-O-glucoside, nicotiflorin, rutin, and narcissin) were found in all varieties; nicotiflorin was predominant, followed by narcissin.

The demand for food is increasing, not only to meet food security for growing populations, but also to provide more nutritious food, rich in good quality proteins and nutraceutical compounds. The amaranth ( Amaranthus hypochondriacus ) plant, in addition to its high nutritive and nutraceutical characteristics, has excellent agronomic features. The objective of the present study was to analyze some physical and proximal-nutritional properties of amaranth seeds obtained from different varieties grown in arid zones and characterize their phenolic acids and flavonoids. Two commercial (Tulyehualco and Nutrisol) and two new (DGETA and Gabriela) varieties of A. hypochondriacus were grown at the Mexican Highlands zone. Tulyehualco and DGETA varieties had higher seed yield of 1475 and 1422 kg ha −1 , respectively, comparable to corn and soybean production in agricultural areas. Gabriela had the highest protein content of 17.3%, but all varieties had an adequate balance of essential amino acids. Polyphenols as rutin (4.0–10.2 μg g −1 flour) and nicotiflorin (7.2–4.8 μg g −1 flour) were detected. Amaranth can be cultivated in arid zones where commercial crops cannot be grown; the seeds besides their well known nutritive characteristics could be a source of phenolic compounds of high antioxidant properties.

Wheat (Triticum aestivum L.) and other cereals produce allelochemicals as natural defense compounds against weeds, fungi, insects and soil-borne diseases. The main benzoxazinoid allelochemical of wheat is 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), bound as beta-glucoside and released upon plant injury. When leached from wheat to soil, DIMBOA is microbially transformed to 6-methoxy-benzoxazolin-2-one (MBOA). Exploiting benzoxazinoids and their degradation products as substitutes for synthetic pesticides depends on knowledge of transformation pathways and kinetics. In an MBOA degradation experiment at a concentration of 2400 nmol g(-1) soil, the previously identified transformation products 2-amino-7-methoxy-phenoxazin-3-one (AMPO) and 2-acetylamino-7-methoxy-phenoxazin-3-one (AAMPO) were quantified. Three different kinetic models were applied to MBOA transformation kinetics; single first-order (SFO), first-order multi-compartment, and double first-order in parallel. SFO proved to be adequate and was subsequently applied to the transformations of MBOA, AMPO and AAMPO. Degradation endpoints, expressed as degradation time (DT), were calculated for MBOA, AMPO and AAMPO to test whether the maximum values for synthetic pesticides set by the European Commission and the Danish Environmental Protection Agency were exceeded. DT(50) values for MBOA and AMPO were 5.4 d and 321.5 d, respectively, and DT(90) values were 18.1 d and 1068 d, respectively. The DT(50) value for AMPO exceeded the maximum value. The persistence, concentrations and toxicity of metabolites such as AMPO should be considered when breeding cereal crops with increased levels of benzoxazinoids.

The Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) procedure was applied and validated for the analysis of neonicotinoid insecticide residues in cocoa bean matrix with high fat and high pigments. Samples employed in the study were fermented and dried beans obtained from major cocoa producing regions in Ghana where neonicotinoids are extensively used. Shells covering the beans were removed and analyzed separately to examine insecticide distribution. Analytes in both matrices were extracted in acetonitrile with sodium acetate and magnesium sulfate salts, cleaned up using a sorbent mixture of primary secondary amine (PSA), C18 and graphitized carbon black (GCB), and quantified using liquid chromatography tandem mass spectrometry (LC–MS/MS). Average recoveries at four levels of fortification ranged from 92 to 111% with relative standard deviation of ≤16% for all analytes. Limits of quantification ranged from 3 to 10 μg/kg for all neonicotinoids. Imidacloprid was the most frequently encountered neonicotinoid and was quantified in more than 10% of deshelled bean and 30% of cocoa shell samples, with concentrations ranging from 11.5 to 35.6 μg/kg and 11.8 to 214 μg/kg in cocoa beans and shells, respectively. The findings from this study suggest a need for greater efficiency in neonicotinoid application, to avoid the build-up of these insecticides to unsafe levels in cocoa beans.

Glyphosate is a widely used non-selective herbicide. Leaching of glyphosate (N-(phosphonomethyl)glycine) and/or its metabolite AMPA (aminomethylphosphonic acid) was studied in four lysimeters, two of them being replicates from a low-tillage field (lysimeter 3 and 4), the other two being replicates from a normal tillage field (lysimeter 5 and 6). In both cases the soil was a sandy loam soil with 13-14% clay. The lysimeters had a surface area of 0.5 m2 and a depth of 110 cm. Lysimeter 3 and 4 were sprayed with a mixture of 14C-labelled glyphosate and unlabelled glyphosate, while lysimeter 5 and 6 were sprayed with unlabelled glyphosate. The spraying took place September 18, 1997. The total amount of glyphosate sprayed onto each lysimeter was 40 mg, corresponding to 0.8 kg active ingredient per ha. The lysimeters were installed in an outdoor system in Research Centre Flakkebjerg and were thus exposed to normal climatic conditions of the area. A mean of 260 l drainage water were collected from lysimeter 3 and 4 and a mean of 375 litres from lysimeter 5 and 6. The mean yearly concentration of leached glyphosate and/or AMPA was significantly below 0.1 microg/l from both sets of lysimeters, and thus no significant difference between the two lysimeter sets was shown. However, in both sets of lysimeters several single findings at concentrations above 0.1 microg/l was seen, which might be due to the leaching of particle-bound compounds. A significant difference between the soil residual concencentrations of AMPA was seen, the higher concentration was found in the set of lysimeter where low-tillage had been practiced and where Round Up had been used several times in the years before sampling of the lysimeter soil.

Benzoxazinoids (BXs) are a group of natural chemical compounds with putative pharmacological and health-protecting properties. BXs were formerly identified in and isolated from selected dicot medicinal plants and young cereal plants. Recently, BXs were found to be present in mature cereal grains and bakery products, such that knowledge about the pharmacological properties of BXs, which until now have unknowingly been consumed through the daily bread and breakfast cereals, have come into new focus. This review discusses published results from in vitro studies and a few human and animal model studies on the health effects and pharmacological responses of various BX compounds. Many of these studies have reported antimicrobial, anticancer, reproductive system stimulatory, central nervous system stimulatory, immunoregulatory, and appetite- and weight-reducing effects of BXs and/or BX derivatives. The health benefits of wholegrain intake may be associated with the solitary and/or overlapping biological effects of fibers, lignans, phenolic acids, alkylresorcinols, BXs, and other bioactive compounds. In the context of BXs as dietary ingredients, further comprehensive investigations are required to understand their biological functions, to elucidate the underlying mechanisms, to explore their potential contribution on the health effects associated with wholegrain consumption, and to examine their potential as functional food ingredients.

Scope Benzoxazinoids, which are natural compounds recently identified in mature whole grain cereals and bakery products, have been suggested to have a range of pharmacological properties and health-protecting effects. There are no published reports concerned with the absorption and metabolism of bioactive benzoxazinoids in humans. Methods and results The absorption, metabolism, and excretion of ten different dietary benzoxazinoids were examined by LC-MS/MS by analyzing plasma and urine from 20 healthy human volunteers after daily intake of 143 μmol of total benzoxazinoids from rye bread and rye buns. The results showed that 2-β-d-glucopyranosyloxy-1,4-benzoxazin-3-one (HBOA-Glc) and its oxidized analog, 2-β-d-glucopyranosyloxy-4-hydroxy-1,4-benzoxazin-3-one (DIBOA-Glc), were the major circulating benzoxazinoids. After consuming a benzoxazinoid diet for 1 week, morning urine contained eight benzoxazinoids with abundant HBOA-Glc (219 nmol × μmol−1 of creatinine). The sulfate and glucuronide conjugates of 2-hydroxy-1,4-benzoxazin-3-one (HBOA) and 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA) were detected in plasma and urine, indicating substantial phase II metabolism. Direct absorption of lactam glycosides, the reduction of hydroxamic acid glycosides, glucuronidation, and sulfation were the main mechanisms of the absorption and metabolism of benzoxazinoids. Conclusion These results indicate that following ingestion in healthy humans, a range of unmetabolized bioactive dietary benzoxazinoids and their sulfate and glucuronide conjugates appear in circulation and urine.

Recently, bioactive benzoxazinoids were discovered in cereal grains and bakery products. In this study, we studied the uptake, distribution, and metabolism of these secondary metabolites using a pig model. Twelve benzoxazinoid compounds and their 4 transformation products were quantified in the pigs' diets and biofluids using high-performance liquid chromatography coupled to electrospray ionization triple quadrupole mass spectrometry. The 2-β-D-glucopyranosyloxy-4-hydroxy-1,4-benzoxazin-3-one (DIBOA-glc) was the most dominant benzoxazinoid (232 nmol/g DM) seconded by the double-hexose derivative of DIBOA (provisionally characterized here as DIBOA-glc-hex) in the rye-based diet. DIBOA-glc (derived from the diet and intestinal deglycosylation of DIBOA-glc-hex) was apparently reduced to 2-β-D-glucopyranosyloxy-1,4-benzoxazin-3-one (HBOA-glc), the most dominant benzoxazinoid in the blood (829 nmol/L). The benzoxazinoid compounds were excreted in the urine, with HBOA-glc (18 μmol/L) as a major metabolite. In this study, we determined for the first time the bioavailability of dietary benzoxazinoids that have high digestibility, distribution, and metabolism in mammals. These findings could be a milestone for the exploitation of healthful and pharmacological properties of benzoxazinoids.

Degradation of biochanin A, 400 μg (1.41 μmol) per gram of dry soil, was studied and the initial degradation products, dihydrobiochanin A, pratensein and genistein were identified and quantified. The maximum concentrations of these compounds were determined as 166 pmol/g dry soil after 2 days of incubation, 175 pmol/g dry soil after 1 day and 1,236 pmol/g dry soil after 1 day, respectively. Biochanin A and the initially formed products degrade too fast to be responsible for clover soil sickness (DT90

The fate of glyphosate and its degradation product aminomethylphosphonic acid (AMPA) was studied in soil. Labeled glyphosate was used to be able to distinguish the measured quantities of glyphosate and AMPA from the background values since the soil was sampled in a field where glyphosate had been used formerly. After addition of labeled glyphosate, the disappearance of glyphosate and the formation and disappearance of AMPA were monitored. The resulting curves were fitted according to a new EU guideline. The best fit of the glyphosate degradation data was obtained using a first-order multi compartment (FOMC) model. DT(50) values of 9 days (glyphosate) and 32 days (AMPA) indicated relatively rapid degradation. After an aging period of 6 months, the leaching risk of each residue was determined by treating the soil with pure water or a phosphate solution (pH 6), to simulate rain over a non-fertilized or fertilized field, respectively. Significantly larger (p0.05) amounts of aged glyphosate and AMPA were extracted from the soil when phosphate solution was used as an extraction agent, compared with pure water. This indicates that the risk of leaching of aged glyphosate and AMPA residues from soil is greater in fertilized soil. The blank soil, to which 252 g glyphosate/ha was applied 21 months before this study, contained 0.81 ng glyphosate/g dry soil and 10.46 ng AMPA/g dry soil at the start of the study. Blank soil samples were used as controls without glyphosate addition. After incubation of the blank soil samples for 6 months, a significantly larger amount of AMPA was extracted from the soil treated with phosphate solution than from that treated with pure water. To determine the degree of uptake of aged glyphosate residues by crops growing in the soil, (14)C-labeled glyphosate was applied to soil 6.5 months prior to sowing rape and barley seeds. After 41 days, 0.006 +/- 0.002% and 0.005 +/- 0.001% of the applied radioactivity was measured in rape and barley, respectively.

Benzoxazinones, such as 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) and 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA), and benzoxazolinones, such as 6-methoxy-2-benzoxazolinone (MBOA) and 2-benzoxazolinone (BOA), are biologically active secondary metabolites found in cereals. Because these compounds could be exploited as part of a strategy for reducing the use of synthetic pesticides, ecotoxicological tests were performed recently. In this paper, the transformation of the compounds in the test environment of the ecotoxicological tests was studied. DIMBOA was degraded and partly transformed to MBOA during the period of ecotoxicological testing of the compounds. During testing of MBOA on Poecilus cupreus test media the analysis showed that at the initial concentrations of 2 and 10 mg kg(-1) no MBOA was left after 45 days of testing, but the metabolite 2-amino-phenoxazin-3-one (AMPO) was formed. During testing of BOA on both Folsomia candida and Poecilus cupreus the more biologically active compound 2-amino-phenoxazin-3-one (APO) was formed. Thus, the ecotoxicological test results on MBOA and BOA were partly due to the microbial transformation of the compounds during the time of testing.

Isoproturon is a herbicide, which was used in Denmark against grass weeds and broad-leaved weeds until 1998. Isoproturon has frequently been detected in ground water monitoring studies. Leaching of isoproturon (N,N-dimethyl-N'-(4-(1-methylethyl)-phenyl)urea) and its metabolites, N'-(4-isopropylphenyl)-N-methylurea and N'-(4-isopropylphenyl)urea was studied in four lysimetres, two of them being replicates from a low-tillage field (lysimeter 3 and 4), the other two being replicates from a normal tillage field (lysimeter 5 and 6). In both cases the soil was a sandy loam soil with 13-14% clay. The lysimetres had a surface area of 0.5 m2 and a depth of 110 cm. Lysimeter 3 and 4 were sprayed with unlabelled isoproturon while lysimeter 5 and 6 was sprayed with a mixture of 14C-labelled and unlabelled isoproturon. The total amount of isoproturon sprayed onto each lysimeter was 63 mg, corresponding to 1.25 kg active ingredient per ha. The lysimeters were sprayed with isoproturon on October 26, 1997. The lysimetres were installed in an outdoor system in Research Centre Flakkebjerg and were thus exposed to normal climatic conditions of the area. A mean of 360 l drainage water were collected from lysimeter 3 and 4 and a mean of 375 litres from lysimeter 5 and 6. Only negligible amounts of isoproturon and its primary metabolites were found in the drainage water samples, and thus no significant difference between the two lysimeter sets was shown. In a total of 82 drainage water samples, evenly distributed between the four lysimetres isoproturon was found in detectable amounts in two samples and N'-(4-isopropylphenyl)urea was found in detectable amounts in two other samples. The detection limit for all the compounds was 0.02 microg/l. 48% and 54% of the added radioactivity were recovered from the upper 10 cm soil layer in lysimeter 5 and 6, respectively, and 17 and 14% from 10-20 cm's depth. By extraction first with an aquatic CaCl2 solution 0.49% of the added radioactivity was extracted from the upper 10 cm layer in lysimeter 5. In the subsequent extraction with acetonitril, 1.19% of the added radioactivity was extracted. In lysimeter 6, upper 10 cm, 0.2% were extracted with water and 0.56% were extracted with acetonitril. Below 10 cm's depth no measurable amounts could be extracted.

An alternative to the use of synthetic pesticides is to exploit the natural defense chemicals produced by cereals. An important class of allelochemicals is cyclic hydroxamic acids and related benzoxazolinones. A prolonged degradation experiment of the allelochemical compound from rye 2-benzoxazolinone (BOA) was carried out for up to 90 d at 15 degrees C at three different concentration levels, 3, 3000, and 30,000 nmol BOA g soil(-1), respectively, in a sandy loam soil. Two main degradation products, 2-amino-(3H)-phenoxazin-3-one (APO) and 2-acetylamino-(3H)-phenoxazin-3-one (AAPO), were identified and quantified by LC-ESI-MS-MS. The half-life of BOA increased with higher levels of BOA added to the soil. Half-lives of BOA, APO, and AAPO were determined by fitting a single first-order model to the degradation data. Half-life of BOA was determined to be 0.6 d in the 3 nmol BOA g soil(-1) treatment. Half-lives of BOA, APO, and AAPO were 3.1, 2.7, and 2.1 d, respectively, in the 3000 nmol BOA g soil(-1) treatment. In the 30,000 nmol BOA g soil(-1) treatment, the half-lives were 31 d for BOA and 45 d for APO. The microbial community structure was not affected by addition of BOA to the soil as investigated by analysis of signature fatty acids. The results suggest that the exploitability of BOA for crop protection is dependent on the existing concentration of BOA in the soil and the timing of incorporation of hydroxamic acid synthesizing crops into the soil.