Validation of a new LC-MS/MS method for the detection and quantification of phenolic metabolites from tomato sauce in biological samples

Tomato is a good source of bioactive molecules such as vitamin C, carotenoids, and phenolic compounds. Up to now, only a few studies have evaluated the bioavailability of phenolic compounds from tomato. This paper presents the optimization of a method for the determination of phenolics in tomato and their metabolites in human urine and plasma after ingestion of tomato sauce. The sample preparation includes a SPE step to obtain cleaner extracts for injection in the LC-MS/MS system. The mean recovery of analytes ranged from 73 to 104% in plasma and from 65 to 106% in urine, the accuracy was between 90.3 and 115.0% in urine and between 85.7 and 115.0% in plasma, and the precision coefficient of variation was <15%. The method allowed detection and quantification limits of 0.5-29 and 2.0-90 ng mL?1 in urine, respectively, and 0.5-30 and 2.0-105 ng mL?1 in plasma, respectively, for the same phenolic compounds.     

Urolithins Are the Main Urinary Microbial-Derived Phenolic Metabolites Discriminating a Moderate Consumption of Nuts in Free-Living Subjects with Diagnosed Metabolic Syndrome

Walnuts ( Juglans regia L.), hazelnuts ( Corylus avellana L.), and almonds ( Prunus dulcis Mill.) are rich sources of ellagitannins and proanthocyanidins. Gut microbiota plays a crucial role in modulating the bioavailability of these high molecular weight polyphenols. However, to date there are no studies evaluating the capacity to produce nut phenolic metabolites in subjects with metabolic syndrome (MetS), a pathology associated with an altered gut bacterial diversity. This study applied a LC-MS targeted approach to analyze the urinary excretion of nut phenolic metabolites in MetS subjects following 12 weeks of nut consumption, compared to sex- and age-matched individuals given a nut-free control diet. Metabolites were targeted in both hydrolyzed and nonhydrolyzed urine by LC-PDA-QqQ-MS/MS analysis, and identification of metabolites lacking available standards was confirmed by LC-ESI-ITD-FT-MS. Ellagitannin-derived urolithins A and B significantly increased after the nut-enriched-diet, urolithins C and D were also detected, and a complex combination of urolithin-conjugated forms was observed in nonhydrolyzed urine, confirming an extensive phase II metabolism after absorption. In contrast, no significant increases in proanthocyanidin microbial metabolites were observed in urine following nut consumption. Because the intestinal microbiota of the subjects in this study could catabolize ellagitannins into a wide range of urolithins, further research is strongly warranted on the in vivo potential of these microbial metabolites in reducing cardiometabolic risk.     

Influence of methanol on the dynamics of the retention and release of cyprodinil by an agricultural soil

The influence of methanol on the adsorption of the fungicide cyprodinil by a crop soil was studied by equilibrium measurements and by determining the retention-release dynamics in a continuous stirred flow tank reactor (CSTR). Equilibrium measurements showed the effective coefficient of partition of cyprodinil between soil and solution, K(dc), decreases linearly as the concentration of methanol in the solution increases until a percentage of 20% is reached. In CSTR experiments, the retention of cyprodinil was found to be almost reversible; up to a 95% of the fungicide was desorbed. The retention-release dynamics showed biphasic behavior and was partially controlled by diffusion. This behavior was reproduced by a model of diffusion into micropores identifying the soil particles as spheres and taking into account both intraparticle nonlinear adsorption and nonlinear adsorption at external surfaces. In all cases, the sorption kinetics was not the limiting step. The main effect of methanol in the retention-release dynamics ended up being based on the changes produced in the adsorption equilibrium. Methanol also increased the effective diffusion coefficient and decreased the mass transfer coefficient. The optimized Freundlich's isotherm coefficients for <5% methanol were lower than those obtained from the batch experiments.     

The role of fat in flavor perception: effect of partition and viscosity in model emulsions

Decreasing the fat content of a food, while maintaining the same aroma content, changes both aroma release (due to partition effects) and the viscosity of the food. To understand the relative contribution of these two factors on flavor perception, a series of flavored emulsions were prepared to control aroma release and viscosity using different aroma, oil, and hydroxypropyl methyl cellulose (HPMC) contents. Samples were formulated to deliver the same aroma-release in vitro and in vivo, and their viscosity was measured using the Kokini oral shear stress parameter. Despite the in vivo aroma release being constant, there were perceptual differences between the samples, and the flavor intensity decreased as in-mouth viscosity increased. For these iso release samples, the Kokini oral shear stress parameter correlated well with the decrease in perception, suggesting that there may be a viscosity stimulus or that the viscosity affects release of tastant and hinders aroma-taste interactions.     

Rapid confirmatory assay for determining 12 sulfonamide antimicrobials in milk and eggs by matrix solid-phase dispersion and liquid chromatography-mass spectrometry

A rapid confirmatory method for determining 12 sulfonamide (SAs) antibacterials in whole milk and eggs is presented. This method is based on the matrix solid-phase dispersion technique with hot water as extractant followed by liquid chromatography (LC)-mass spectrometry (MS). The LC-MS instrument was equipped with an electrospray ion source and a single quadrupole. After 4 mL of a milk sample containing the analytes had been deposited on sand (crystobalite), this material was packed into an extraction cell. SAs were extracted by flowing 4 mL of water through the cell heated at 75 degrees C. With some modifications, this procedure was applied also to eggs. After pH adjustment and filtration, 0.5 mL of the final extracts was then injected into the LC column. MS data acquisition was performed in the positive-ion mode and by monitoring at least three ions for each target compound. The in-source collision-induced dissociation process produced confirmatory ions. At the 50 ng/g level, recovery of the analytes in milk and eggs was 77-92% with relative standard deviations ranging between 1 and 11%. Estimated limits of quantification (S/N = 10) were 1-3 ng/g of SAs in milk and 2-6 ng/g in eggs. With both matrices, attempts to reduce the analysis time by using a short chromatographic run time caused severe ion signal suppression for the early-eluted SAs. This effect was traced to competition effects by polar endogenous coextractives, maybe proteinaceous species, which are eluted in the first part of the chromatographic run. This unwelcome effect was almost completely removed by simply adopting more selective chromatographic conditions.     

The Maximum Incremental Social Tolerable Irreversible Costs (MISTICs) and other benefits and costs of introducing transgenic maize in the EU-15

The decision to release a new transgenic crop variety for planting in the European Union (EU) is a decision under irreversibility and uncertainty. We use a real option model to assess the ex-ante incremental benefits and costs of the decision to release Bt maize and HT maize in the EU-15 member states. The analysis uses Eurostat data for modelling the benefits and costs of non-transgenic maize using partial equilibrium models. The farm-level benefits and costs of Bt maize and HT maize are derived from field trials conducted within the EU-funded ECOGEN project in combination with secondary data sources. Adoption curves, hurdle rates and Maximum Incremental Social Tolerable Irreversible Costs (MISTICs) are calculated at country level for selected EU-15 member states. In general, the results show that the MISTICs on a per capita level are very small confirming previous results calculated in values for the year 1995. The MISTICs per farm are much larger. This indicates a problem for decision makers.     

Phytotoxicity of natural soils using physiological and biochemical endpoints reveals confounding factors: can a weight of evidence tackle uncertainty?

Purpose

Standard assays for phytotoxicity provide a reductionist view on the performance of plants under toxic stress. To address two of the most prominent issues in plant toxicity studies, our aims were (1) to assess how well physiological and biochemical parameters complement standard toxicological endpoints when testing natural soils and (2) to assess the suitability of three commonly used control soils as comparative references.

Material and methods

We compared the performance of Zea mays and Helianthus annuus in three control soils (artificial Organisation for Economic Co-operation and Development (OECD) soil, standard LUFA 2.2 soil, and turf-perlite) against three natural soils representing a gradient of contamination (from a deactivated uranium mine). Standard endpoints (emergence and biomass) were estimated, along with pigment content, photosynthetic parameters, cellular injury, and proline content.

Results and discussion

The toxicological profile of natural soils was highly dependent on the control soil used as reference; also, plant physiological performance was influenced by the soils’ properties. We discuss the need to interpret and combine multiple lines of evidence as a way to increase the degree of confidence one classifies soils based on their ecotoxicity, and this is where the integration of physiological and biochemical parameters bring added value.

Conclusions

When facing large variability in soil characteristics, it is best to collect and integrate as much information possible to strengthen conclusions about phytotoxicity of natural soils. Obviously, this refutes reductionist views and places the final conclusion in the hands of expert judgment.

     

Use of the chloroplast gene ycf1 for the genetic differentiation of pine nuts obtained from consumers experiencing dysgeusia

Pine nuts are a part of traditional cooking in many parts of the world and have seen a significant increase in availability/use in the United States over the past 10 years. The U.S. Food and Drug Administration (US FDA) field offices received 411 complaints from U.S. consumers over the past three years regarding taste disturbances following the consumption of pine nuts. Using analysis of fatty acids by gas chromatography with flame ionization detection, previous reports have implicated nuts from Pinus armandii (Armand Pine) as the causative species for similar taste disturbances. This method was found to provide insufficient species resolution to link FDA consumer complaint samples to a single species of pine, particularly when samples contained species mixtures of pine nuts. Here we describe a DNA based method for differentiating pine nut samples using the ycf1 chloroplast gene. Although the exact cause of pine nut associated dysgeusia is still not known, we found that 15 of 15 samples from consumer complaints contained at least some Pinus armandii, confirming the apparent association of this species with taste disturbances.