Simultaneous determination of four biogenic and three volatile amines in anchovy by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry

A new method has been developed and validated for the simultaneous determination of four biogenic (putrescine, cadaverine, histamine, and tyramine) and three volatile amines (trimethylamine, triethylamine, and tripropylamine) in anchovy. Separation and determination of the selected compounds were carried out by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS), using an electrospray ionization source (ESI) in positive ion mode. Chromatographic separation was carried out using an aqueous solution of formic acid (0.1%) and methanol as mobile phase in gradient mode. The method was validated, and mean recoveries were evaluated at three concentration levels (75, 150, and 250 μg/kg), ranging from 70 to 110% at the three levels assayed. Intra- and interday precision, expressed as relative standard deviation (RSD), were lower than 15% and 20%, respectively. Limits of quantitation (LOQs) were 25 μg/kg for all cases, except for that of TMA, which was set at 60 μg/kg. The developed procedure was applied to determine the target compounds in anchovy samples stored during 7 days at 4 °C, observing the increasing in the concentration of these compounds at longer storage time.     

Chemical conversion of phenylethylamine into phenylacetaldehyde by carbonyl-amine reactions in model systems

The chemical conversion of phenylethylamine into phenylacetaldehyde in the presence of lipid oxidation products (LOPs) was studied to investigate the possibility that biogenic amines can be converted into Strecker aldehydes upon processing. Model systems of phenylethylamine and methyl 13-hydroperoxyoctadeca-9,11-dienoate (HP), 2,4-decadienal (DD), 4,5-epoxy-2-heptenal (EH), 4,5-epoxy-2-decenal (ED), 4-oxo-2-hexenal (OH), 4-oxo-2-nonenal (ON), or 4-hydroxy-2-nonenal (HN) were heated for 1 h at 180 °C and pH 3. Although HN and EH did not produce more phenylacetaldehyde than when phenylethylamine was heated alone, all other lipid oxidation products assayed increased the amount of phenylacetaldehyde produced by 300-900%, with ON being the most reactive compound for this reaction. The reaction was mainly produced at acidic pH values (<6) and was dependent upon the concentration of the LOPs involved, and the phenylacetaldehyde produced increased linearly as a function of the time and temperature. The E(a) values for the reactions between phenylethylamine and DD and ON were 54.8 and 53.8 kJ/mol, respectively. The reaction is proposed to take place by the formation of an imine between the phenylethylamine and the LOPs, which is later converted into another imine by an electronic rearrangement. This new imine is the origin of phenylacetaldehyde by hydrolysis. These results show a new pathway for Strecker aldehyde formation. This route provides a potential way to reduce biogenic amine content in foods when they can be thermally processed before consumption.     

Thermal inactivation kinetics of recombinant proteins of the lipoxygenase pathway related to the synthesis of virgin olive oil volatile compounds

The aim of this work was to characterize the thermal inactivation parameters of recombinant proteins related to the biosynthesis of virgin olive oil (VOO) volatile compounds through the lipoxygenase (LOX) pathway. Three purified LOX isoforms (Oep2LOX1, Oep1LOX2, and Oep2LOX2) and a hydroperoxide lyase (HPL) protein (OepHPL) were studied. According to their thermal inactivation parameters, recombinant Oep1LOX2 and Oep2LOX2 could be identified as the two LOX isoforms active in olive fruit crude preparations responsible for the synthesis of 13-hydroperoxides, the main substrates for the synthesis of VOO volatile compounds. Recombinant Oep2LOX1 displayed a low thermal stability, which suggests a weak actuation during the oil extraction process considering the current thermal conditions of this industrial process. In addition, recombinant OepHPL could be identified as the HPL activity in crude preparations. The thermal stability was the highest among the recombinant proteins studied, which suggests that HPL activity is not a limiting factor for the synthesis of VOO volatile compounds.     

Impact of must sugar reduction by membrane applications on volatile composition of Verdejo wines

Climate changes are inducing increased sugar levels of must, which produces negative effects on wine quality, as unbalanced wines with high degrees of alcohol. So, effective strategies to control the increase of sugar levels in must have been studied. One of them is the use of a membrane process, and this is applied in this work. The sugar level of white must from Verdejo (Vitis vinifera variety) was reduced using diverse membrane processes, and the effect of this fact on the volatile composition of the corresponding wines is studied. The study was carried out during three consecutive vintages. An important impact of the reduction of sugar levels of must on the volatile composition of the obtained wines was detected, which was due to some retention phenomena of aromatic and precursor compounds. To minimize the volatile composition modifications, an appropriate selection of the nanofiltration membrane must be done.     

Hydrolysis of Chicoric and Caftaric Acids with Esterases and Lactobacillus johnsonii in Vitro and in a Gastrointestinal Model

Chicoric acid (ChA) and caftaric acid (CafA) were identified as bioactive components of chicory and have been ascribed a number of health benefits. This study investigated the hydrolysis of ChA and CafA with enzymes and a probiotic bacterium Lactobacillus johnsonii (La1). Esterase from Aspergillus japonicus (24 U/mg) hydrolyzed 100% of ChA (5 mM) and CafA (5 mM) after 3 h, at pH 7.0 and 37 °C. Under the same reaction conditions, 100% hydrolysis of ChA and CafA was achieved with a spray-dried preparation of La1. The addition of La1 (100 mg/mL, 3.3 E9 cfu/g) to CafA solution in a gastrointestinal model (GI model) resulted in 65% hydrolysis of CafA. This model simulates the physicochemical conditions of the human gastrointestinal tract. No hydrolysis of CafA was observed after passage through the GI model in the absence of La1. The results of this study support the hypothesis that ChA and CafA are degraded by gut microflora before absorption and metabolization.     

Enhanced Dechlorination of Carbon Tetrachloride by Immobilized Fulvic Acids on Alumina Particles

Fulvic acids (FA) were immobilized on alumina particles in order to evaluate their catalytic effect as solid-phase redox mediator (RM) during the reductive dechlorination of carbon tetrachloride (CT) by anaerobic sludge. FA were extracted from three different soil samples and two commercial composts. Electron carrying capacity (ECC) was determined in all FA samples in order to select the appropriate source of redox-mediating compounds for CT dechlorination. TiO₂, Al(OH)₃, and γ-Al₂O₃ particles were tested as immobilizing materials for the extracted FA. FA extracted from a temperate pine forest soil showed the highest ECC (291.72 μmol g⁻¹). The highest adsorption capacity of FA, measured as total organic carbon (TOC), was achieved by alumina (γ-Al₂O₃) particles (12 mg TOC-FA g⁻¹). Results suggest that the transfer of electrons rather than their microbial generation through glucose fermentation was the rate-limiting factor during dechlorination of CT. Immobilized FA increase up to 10.4-fold the rate of CT dechlorination as compared with the control lacking FA. Immobilization of FA on alumina particles was very stable, and spectrophotometric screening did not detect any detachment of FA during dechlorination of CT, thus confirming that the enhanced dechlorination achieved could exclusively be linked to the redox-mediating capacity of immobilized FA. The present study constitutes the first demonstration that immobilized FA on alumina particles could serve as a solid-phase RM in dechlorination reactions.     

Assessment of s-Triazine Catabolic Potential in Soil Bacterial Isolates Applying atz Genes as Functional Biomarkers

Fluorescence in situ hybridization (FISH) technique and qPCR analyses, targeting atz genes, were applied to detect the presence of simazine-degrading bacteria in an agricultural soil with a history of herbicide application. atzB-targeted bacteria detected by FISH represented 5% of total soil bacteria with potential capability to metabolize the herbicide. The soil natural attenuation capacity was confirmed in soil microcosms by measuring simazine degradation. Moreover, four bacterial strains were isolated from the soil and identified as Acinetobacter lwoffii, Pseudomonas putida, Rhizobium sp. and Pseudomonas sp. The isolates were able to grow using different s-triazine compounds and related metabolites as the sole carbon source. Growth parameters in presence of simazine were calculated using the Gompertz model. Rhizobium sp. showed the highest simazine degradation (71.2%) and mineralization (38.7%) rates, whereas the lowest values were found to A. lwoffii??50.4% of degradation and 22.4% of mineralization. Results from qPCR analyses of atzA, atzB and atzC genes revealed their presence in Rhizobium sp. and A. lwoffii, being atzB and atzC the most abundant functional genes. Rhizobium sp. showed a higher amount of the three biomarkers compared to A. lwoffii: the atzA, atzB and atzC gene copy number per microlitre were, respectively, 10¹, 10² and 10³-fold higher in the former. Therefore the proposed molecular approaches based on the use of atz genes as biomarkers can be considered as useful tools to evaluate the presence and potential capability of degrading-s-triazines soil microorganisms.     

Aerobic Biological Treatment of Chestnut Processing Wastewater

Chestnut agro-industrial companies consume a high volume of water for washing and processing fruit, generating a large volume of wastewater. This work studied the biodegradation of chestnut processing wastewater through aerobic assays, varying substrate, and biomass concentrations. In general, this wastewater presents a good biodegradability, especially in experiments with relatively low chemical oxygen demand (COD) (0.4 and 0.6?g O₂ L^?1) allowing a COD removal of 85?C90?%. The best results were obtained in the reactor initially loaded with 2?g?L^?1 of biomass and 0.4 or 0.6?g O₂ L^?1 of COD. These experiments also showed high COD removal rates: 4.25 and 3.88?g COD g^?1 volatile suspended solids (VSS) h^?1, respectively. The sedimentation rate, evaluated for different initial values of biomass (1, 2, and 3?g?L^?1), always presented higher values in the experiments with 2 and 3?g?L^?1 of biomass, regardless of the initial COD value used. After comparing different kinetic models (Monod, Contois, and Haldane), it was observed that the Haldane inhibition model satisfactorily describes the COD biodegradation. AQUASIM software allowed calculating the kinetic constant ranges: K _s, 1.59?C6.99?g COD L^?1; ?? _max, 25?C40?g COD g^?1 VSS day^?1; and K _i values, 0.07?C0.11. These kinetic constants corresponds to maximum rates (??*) between 1.48 and 4.25?g COD g^?1 VSS day^?1 for substrate concentrations (S*) from 0.38 to 0.88?g COD L^?1.     

An Exploratory Study of Particulate PAHs in Low-Polluted Urban and Rural Areas of Southwest Spain: Concentrations, Source Assignment, Seasonal Variation and Correlations with Other Air Pollutants

A set of 72?PM₁₀ samples from low-polluted urban and rural locations belonging to the regional air monitoring network of Extremadura (Spain) were collected in a 1?year sampling period. Sample pre-treatment and analytical determination by gas chromatography?Cion trap mass spectrometry were optimised and validated for the analysis of the priority 16 US Environmental Protection Agency polycyclic aromatic hydrocarbons (PAHs). The influence of meteorological conditions (temperature, relative humidity and solar radiation) and other atmospheric pollutants (O₃, NO₂, SO₂, PM₁₀) has been covered in detail and Pearson correlation test were used for this purpose. Spatial distribution of particulate PAHs was evaluated and the comparison with other European sites was also established. Possible emission sources were identified and assigned by using molecular diagnostic criteria.     

Losing a staging area: Eastward redistribution of Afro-Eurasian ruffs is associated with deteriorating fuelling conditions along the western flyway

The fuelling performance of long-distance migrants at staging areas indicates local conditions and determines the viability of migration routes. Here we present a first case study where long-term fuelling performance was documented along two migration routes with differential population trends. Ruffs (Philomachus pugnax) are shorebirds of inland freshwater wetlands that migrate from the sub-Saharan wintering grounds, via Europe, to the northern Eurasian breeding grounds. Assessments from 2001 to 2008 of fuelling during northward migration at the major western and eastern staging site revealed that daily mass gain rates steeply declined across years in the grasslands for dairy production in Friesland, The Netherlands, and remained constant in the Pripyat floodplains in Belarus, 1500 km further east. Migrants in Friesland decreased from 2001 to 2010 by 66%, amounting to a loss of 21,000 individuals when counts were adjusted for length of stay as determined by resightings. In the same period numbers in Pripyat increased by 12,000. Ruffs individually ringed in Friesland were resighted in subsequent springs at increasingly eastern sites including Pripyat. Our results corroborate published evidence for an eastward redistribution of Arctic breeding ruffs and suggest that the decreasing fuelling rates in the westernmost staging area contribute to this redistribution. The shift implies that responses occur within a single generation. The hypothesis that the choice of route during northward migration may be driven by food availability can now be tested by creating greater areas of wet grasslands in Friesland. When local staging conditions improve we predict that ruffs will make the reverse shift.