Ledicia Rey-Salgueiro

Effects of toasting procedures on the levels of polycyclic aromatic hydrocarbons in toasted bread

Some polycyclic aromatic hydrocarbons (PAHs), particularly those with a high molecular weight, have been classified as probably carcinogens to humans by the International Agency for Research on Cancer (IARC). The significance of the determination of PAHs is reflected by the special attention of the European Union, which is paying to regulate the maximum allowed levels of PAHs in foodstuffs such as smoked foods. Like other thermally processed foodstuffs, toasted bread can contain these carcinogenic chemicals, not only due to a contamination at source but also during toasting. In order to check PAHs generated from toasting in sandwich bread, several treatment conditions were evaluated: direct toasting (flame-toasting, coal-grilling or gas oven-toasting) or indirect toasting (electric oven-toasting). PAHs were extracted by solid-liquid extraction (SLE) and determined by liquid chromatography with fluorescence detection (LC-FD). Based on the results, the used toasted technique would strongly affect in PAH levels in the final product. No samples obtained by electric oven and toaster were polluted; otherwise the samples toasted by charcoal and flame grilling presented very important levels. Up to 350μg/kg of total PAHs were detected in toasted samples by wood flame. Differences between different ways of toasting could be ascribed to deposition of PAHs from smoke. Finally, several commercial toasted samples of bread were tested to determine PAHs. Overall, the PAH levels were very low. Benzo[a]pyrene ranged from no detectable to 0.23μg/kg.

Ultrasound-assisted emulsification–microextraction for the determination of phenolic compounds in olive oils

A reliable, sensitive and effective method based on ultrasound-assisted emulsification-microextraction (USAEME) coupled to HPLC-DAD has been developed to identify and quantify several target phenolic compounds from extra virgin olive oils (EVOO). This approach is based on the emulsification of a microvolume of polar organic extractant in a non-polar liquid sample by ultrasound radiation and further separation of both liquid phases by centrifugation. The percentage of methanol/water (v/v) in the extractant, the volume of extractant, and the extraction time as three effective parameters on the extraction were optimised by a central composite design (Box-Behnken response surface) method. The optimised method presented recoveries in EVOO between 91% and 115% for the target analytes (except vanillin with 65%) and a satisfactory precision with relative standard deviations (RSD%) lower than 8.4% for repeatability and reproducibility. The method showed good linearity and limits of detection and quantification were in the range 0.001-0.14 and 0.004-0.47mg/kg, respectively. After method validation, it was successfully applied to the analysis of three EVOO samples. All target compounds were detected in all analysed samples. Tyrosol and hydroxytyrosol were the major phenolic compounds, followed by pinoresinol and luteolin.

Comparative performance of extraction strategies for polycyclic aromatic hydrocarbons in peats.

The assessment of historical trends in atmospheric deposition of organic contaminants by using peat samples has been reported on several occasions because these samples represent an almost ideal medium for recording temporal changes in organic contaminant deposition rates. The determination of polycyclic aromatic hydrocarbons (PAHs) in peat samples is complicated due to the high content of organic matter in peat, which affects both extraction efficiency and analytical quality. A rapid and simple method is proposed for the determination of 10 US Environmental Protection Agency indicator PAHs in complex matrices such as peat. This article reviews and addresses the most relevant analytical methods for determining PAHs in peat. We discuss and critically evaluate three different extraction procedures, such as ultrasound-assisted solvent extraction (UASE), shaking and pressurized liquid extraction (PLE). Clean-up of extracts was performed by solid-phase extraction using silica cartridges. Detection of the selected PAHs was carried out by high-performance liquid chromatography coupled with fluorescence detection for determination. Optimization of the variables affecting extraction by the selected extraction techniques was conducted, concluding that the UASE extraction method using hexane:dichloromethane (80:20) as extractant was robust enough to determine the selected PAHs in peat samples with estimated quantification limits between 0.050 and 3.5 microg/kg depending on the PAH. UASE did not demand sophisticated equipment and long extraction times. PLE involved sophisticated equipment and showed important variations in the results. The method proposed was applied to the determination of PAHs in peat samples from Xistral Mountains (Galicia, Spain).

Pre-industrial accumulation of anthropogenic polycyclic aromatic hydrocarbons found in a blanket bog of the Iberian Peninsula.

Studies on the temporal deposition of polycyclic aromatic hydrocarbons (PAHs) in peatlands are scarce, and none have been carried out in the Iberian Peninsula. To address this gap, ten PAHs were determined in a short peat core (spanning the last 1000 years) sampled in NW Iberian Peninsula, by HPLC-fluorescence. Fluoranthene, pyrene, benzo[b]fluoranthene and indeno[1,2,3-cd]pyrene predominated in the upper layers (10 cm), whereas fluoranthene and pyrene were the most abundant in the lower layers (40 cm), which showed an absence of high molecular weight PAHs (benzo[ghi]perylene and Indeno[1,2,3-cd]pyrene). Although increased PAH contents have been detected since 1700 A.D., coinciding with the beginning of the Metallurgical and Industrial Revolution, high levels of fluoranthene and pyrene were present in peat samples dating back to the 12th century A.D. The results suggest that changes in sources, type of emission (global or local) and transport could be responsible for the different PAH content and composition of the peat core. These changes are consistent with the history of the use of natural resources in the NW of the Iberian Peninsula.

The use of manures for detection and quantification of polycyclic aromatic hydrocarbons and 3-hydroxybenzo[a]pyrene in animal husbandry

PAHs from a polluted atmosphere are generally transferred to plants by particle-phase deposition on the waxy leaf cuticle or by uptake in the gas phase through stomata. Thus, they are also present in ingredients for animal feed. Generally, toxic substances are metabolized before or after absorption through the intestinal tract. This is the case of 3-hydroxybenzo[a]pyrene, which can be found free or in its glucuronide and/or sulphate conjugate forms. This article develops a procedure to monitor the carry-over of PAHs from feed to food of animal origin based on the analysis of animal manure. Eleven PAHs and 3-hydroxybenzo[a]pyrene were measured in four animal manures (cow, horse, rabbit and pig) by liquid chromatography with fluorescence detection. benzo[a]pyrene, benzo[b]fluoranthene and benzo[k]fluoranthene were found in all the selected samples, but their total levels were not alarming (benzo[a]pyrene equivalents (microg/kg)<3.1). 3-hydroxybenzo[a]pyrene was quantified in all the samples exempting rabbit manure. The highest total PAH levels were detected in cow manure (9.0 microg/kg), while the highest 3-OH-B[a]P level was determined in horse samples (13 microg/kg).

Training of panellists for the sensory control of bottled natural mineral water in connection with water chemical properties.

As bottled mineral water market is increasing in the world (especially in emergent and developed countries), the development of a simple protocol to train a panel to evaluate sensory properties would be a useful tool for natural drinking water industry. A sensory protocol was developed to evaluate bottled natural mineral water (17 still and 10 carbonated trademarks). The tasting questionnaire included 13 attributes for still water plus overall impression and they were sorted by: colour hues, transparency and brightness, odour/aroma and taste/flavour/texture and 2 more for carbonated waters (bubbles and effervescence). The training lasted two months with, at least, 10 sessions, was adequate to evaluate bottled natural mineral water. To confirm the efficiency of the sensory training procedure two sensory groups formed the whole panel. One trained panel (6 persons) and one professional panel (6 sommeliers) and both participated simultaneously in the water tasting evaluation of 3 sample lots. Similar average scores obtained from trained and professional judges, with the same water trademarks, confirmed the usefulness of the training protocol. The differences obtained for trained panel in the first lot confirm the necessity to train always before a sensory procedure. A sensory water wheel is proposed to guide the training in bottled mineral water used for drinking, in connection with their chemical mineral content.

Liquid chromatography-mass spectrometry method development for monitoring stress-related corticosteroids levels in pig saliva.

Biochemical response stressors results in an increase of adrenocortical activity. Before knowing the corticosteroid levels in saliva in a stressful situation, baselines salivary levels should be established. A method for simultaneous determination of five corticosteroids was developed, validated and applied to pig saliva at farms. The method employs solid-phase extraction (SPE) coupled with clean-up extraction step using silica cartridge in the same step followed by liquid chromatography/tandem mass spectrometry (LC-MS/MS), using electrospray ionization (ESI) in positive mode. The overall method quantification limits range from 0.050 to 0.30μg/L for the enrichment of 1.0mL saliva samples and analyte recoveries are between 60 and 90% (RSD<11%). Some factors studied were: pig sex, breeds, and time at farm. The analytical method clearly shows that CRL and CRS levels of, respectively, 3.0 and 4.0μg/L in saliva can be indicative of maxima non-stress levels in different pig breeds at farm.

Determination of kinetic bioconcentration in mussels after short term exposure to polycyclic aromatic hydrocarbons

The kinetic bioconcentration of N-heterocyclic aromatic hydrocarbons and polycyclic aromatic hydrocarbons in mussels (Mytilus galloprovincialis) after short waterborne exposure was studied. Benzo[a]pyrene (BaP), its analogue azaarene 10-azabenzo[a]pyrene (AzaBaP), and their mixture (Mix), were selected to monitor the changes in water concentrations over three days. Decay of both PAHs concentrations in water after 24 h of waterborne exposure to mussels at levels of 10 and 100 μg/L follows a first order kinetic with half-lives of 4–5 h, with residual levels of PAHs below 7%. While steady-state scenarios are well studied, there is a lack of information of what happens under non-steady-state conditions, the main purpose of our paper. A synergistic bioconcentration of the mixture was found (around 800 in the mix vs. around 400 for individual PAHs at 100 μg/L of waterborne exposure). It could be explained by the following reasons. The most polar AzaBaP does not compete with the most non-polar BaP for the same tissue compartments. Whereas BaP aggregate in hydrophobic areas, AzaBaP can also do in hydrophilic areas. Moreover, a chance for complex formation between them by charge-transfer stabilization mechanisms could make possible a higher bioaccumulation as a mixture. Instead, toxicological results suggest an additive behaviour in the mixture performance, dominated by BaP, which is the key PAH controlling phase I metabolization in mussels, since is approx. three times more toxic. These experiments provide useful indications for a rapid assessment of PAHs kinetic bioconcentration in mussels.

Meat quality in relation to swine well-being after transport and during lairage at the slaughterhouse

Cortisol and corticosterone in saliva were evaluated as pig stress biomarkers, using pig genotype (Duroc, L62 or Pietrain) and lairage time in the slaughterhouse (0, 2.0, 4.0 or 6.0 h) as controlled variables. Although some pigs were found to be carriers of stress susceptibility, all were healthy heterozygous individuals. Pre-slaughter transport increased cortisol levels in saliva above 3.0 μg/L (medium stress), and 4.0 h of lairage in the slaughterhouse raised them above 6.0 μg/L, whereas corticosterone concentrations exceeded 4 μg/L, which are suggestive of high stress. The highest cortisol levels were detected in the Duroc genotype. Other factors such as food deprivation, background noise, the presence of a large number of animals waiting to be slaughtered, mixing with unfamiliar animals or recent mixing of genders may also influence stress. Corticosterone proved a reliable indicator of high stress only. Meat quality from the pig breeds studied was not affected by lairage in the slaughterhouse for up to 6.0 h.