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Carmines (E120) are used worldwide as natural food colouring agents of animal origin, with a widespread application, including yoghurts. Despite being considered safe for human health, carmines are known to cause allergic reactions. Our goal was to evaluate the presence of carmines in different yoghurts with a label declaration of E120, purchased in Portugal, and, for the first time, to assess the human risk. The analytical methodology, recommended by JECFA, was based on acid hydrolysis of the samples followed by spectrophotometric UV-Vis analysis at 494 nm. This methodology allowed for a limit of quantification (LOQ) of 39.0 mg/kg and recovery rates higher than 97.7%. All the samples had carmines at levels above the LOQ, ranging between 43.8 and 193.8 mg/kg, with an average of 125.2 ± 34.5 mg/kg. In total, 8 (26.7%) samples exceeded the European Union (EU) maximum permitted level (MPL) established for carmines in this foodstuff, 150 mg/kg. Solid yoghurts presented higher average levels, 137.2 mg/kg, when compared to liquid samples, 107.2 mg/kg, with a significant statistical difference (

The use of glyphosate has been increasing over the years, making it one of the most consumed herbicides in the world. Although children are considered a vulnerable population, only four previous published studies determined glyphosate in the urine of non-occupationally exposed children. The paucity of epidemiological data and biomonitoring surveys are considered major gaps, that hinder the implementation of science driven policies in the protection of public health. The aim of the present study was to determine glyphosate in the urine of 41 Portuguese children (2-13 years old) and identify potential determinants of exposure. Glyphosate was detected in 95.1% of the samples (1.77 ± 0.86 μg/L), up to a maximum value of 4.35 μg/L. Glyphosate concentrations were higher in the urine of children aged 7-9 years, living near agricultural areas (1 km), with a higher percentage of consumption of home-produced foods, and whose parents applied herbicides in the backyard. Risk assessment revealed an exposure representing 1-5.58% of the established Acceptable Daily Intake (ADI) of glyphosate (0.5 mg/kg bw/day). The results should be further analyzed considering the age of the participants, for which no adjusted ADI exists. This was the first published report of glyphosate exposure in the urine of Portuguese children.

Artificial sweeteners such as acesulfame-potassium (ACE-K). saccharin (SAC), and aspartame (ASP), play a vital role in the food industry. Despite apparent benefits, several studies report adverse reactions on consumers. EFSA is currently re-evaluating several individual sweeteners, including ASP, Ace-K and SAC that shall be completed by 31 December 2020. Our goal was to evaluate, by liquid chromatography with ultraviolet detection (LC-UV), their levels in non-alcoholic beverages from the Portuguese market (soft drinks, sport drinks, energy drinks, and nectars), and estimate the exposure of the Portuguese adolescent/adult population. ACE-K was found in 85.7% of the samples, in mean levels of 97.4 mg/L. ASP and SAC, detected in 37.5% and 33.9% of the samples, were found in averages of 161.5 and 55.0 mg/L, respectively. The maximum limits set for ASP were exceeded in one energy drink. SAC exceeded the maximum permitted level (MPL) level in 3 soft drinks based on tea extracts. In 4 samples SAC was found but not mentioned in the label, and in one of these samples exceeded the MPL. The estimated daily intake (EDI), calculated in mg per kilogram of body weight per day, of ACE-K, SAC and ASP was found to be below the acceptable daily intake (ADI), both for Portuguese adolescents and adult population. Traditional soft drinks accounted for the highest EDIs representing 1.8%, 2.3% and 0.6% for ACE-K, SAC and ASP, respectively, of the ADI. Considering the previous report in Portugal it is observed that the population is currently exposed to a greater amount of artificial sweeteners, which may indicate a trend for more artificial sweeteners' usage in order to reduce sugar intake.

Ochratoxin A (OTA) is produced by fungi present in several agricultural products with much relevance to food safety. Since this mycotoxin is widely found in cereals, beer has a potential contamination risk. Therefore, it was deemed essential to quantify, for the first time, the levels of OTA in beer, a cereal-based product that is marketed in Portugal, as well as to calculate the human estimated weekly intake (EWI) and risk assessment. A total of 85 samples were analyzed through immunoaffinity clean-up, followed by liquid chromatography-fluorescence detection (LC-FD). This analytical methodology allowed a limit of quantification (LOQ) of 0.43 &micro, g/L. The results showed that 10.6% were contaminated at levels ranging between &lt, LOQ and 11.25 &micro, g/L, with an average of 3.14 &plusmn, 4.09 &micro, g/L. Samples of industrial production presented lower incidence and contamination levels than homemade and craft beers. On what concerns human risk, the calculated EWI was significantly lower than the tolerable weekly intake (TWI). However, in the worst case scenario, based on a high concentration, the rate EWI/TWI was 138.01%.

Mycotoxins are secondary metabolites of fungi that have toxic effects on both humans and animals. Aflatoxin M1 (AFM1) which can be transmitted to newborns via breast milk, is a hydrolyzed metabolite of Aflatoxin B1 (AFB1) that is ingested along with contaminated food. AFM1 is classified as “possibly carcinogenic agent for Human” (group 2B IARC). Despite the potential hazard of AFM1 to breastfed babies, there is a lack of studies of exposure evaluation in Europe and a complete absence of reports in Portugal. The present study aimed to determine the occurrence of AFM1 in maternal milk and the degree of exposure of infants to this toxin. The correlation between the concentration of AFM1 and basic socio-demographic factors and the consumption of certain categories of food was also aimed. Thus 67 milk samples from nursing mothers living in Portugal were collected, between 2015 and 2016, and analyzed using a competitive commercial ELISA kit, in order to determine the presence of AFM1. Twenty-two samples (32.8%) contained levels of AFM1 above the detection limit (5 ng/L), ranging between 5.1 and 10.6 ng/L (7.4 ± 1.9 ng/L). In the observed exposure pattern, AFM1-positive milk samples were associated with summer collection, lower mother's educational level, early lactation phase and the maternal consumption of rice and chocolate. No other studied determinants, whether socio-demographic (age, weight, height, number of children, characteristics of breastfeeding, the infants' weight) or dietary (frequency of food consumption) showed a significant statistical influence. Assessment of AFM1 estimated daily intake (EDI) revealed a higher exposure of younger babies ( Thus the results of this study suggest the need to reinforce surveillance of AFB1 occurrence in food, as a protective and control measure, not only for adults, but ultimately for lactating infants exposed through maternal breast milk. Although AFM1 presents an inferior carcinogenic potency, it is noteworthy that when compared with adults, infants feature a lower capacity of carcinogen biotransformation, a fairly restricted diet and a higher consumption in relation to body weight.

Selective serotonin reuptake inhibitors (SSRIs) are among the most prescribed antidepressants nation and worldwide. Their presence in the aquatic environment is known, causing growing pressures in heavily populated coastal areas. For instance, disruption of invertebrate endocrine systems has been described. Since discrete seawater sampling may not be suitable to evaluate such environmental risk, we aimed to assess SSRIs and metabolites in wild Mytilus galloprovincialis, and assess risk to human health. A total of 1225 mussels were sampled during one year along the Portuguese Atlantic coast. Each freeze-dried pool of 25 specimens was analyzed, in duplicate, by solid phase extraction (SPE) and liquid chromatography with tandem mass spectrometry (LC-MSn). Approximately 70% of the samples were contaminated with at least one, and up to 4 from the 7 targeted compounds. The cumulative SSRIs and metabolites content in mussels ranged between 3.17 and 33.93ngg-1 dry weight, in a mean level of 14.68±9.69ngg-1. Norfluoxetine was the most recurring SSRI and the one with the highest mean concentration, 13.52ngg-1. Although some differences were found between different sampling periods and locations, those were not statistically significant (p>0.05) and no correlation was found between wet weight and mean contamination levels. Regarding risk to human health following consumption of these filter-feeders, when considering the average sample contamination of positive samples and the average Portuguese and European consumption, the ratio between the estimated daily intake (EDI) and the acceptable daily intake (ADI) ranged from 0.45 to 5.50%, and from 1.60 to 19.41%, respectively. Therefore, although the resultant risk is lower for the Portuguese consumer than for the average EU consumer, apparently, no appreciable risk to human health is expected.

Pharmaceuticals occurrence and environmental risk assessment were assessed in Portuguese surface waters, evaluating the impact of wastewater treatment plants (WWTPs) and river flow rates. Twenty three pharmaceuticals from 6 therapeutic groups, including metabolites and 1 transformation product, were analysed in 72 samples collected from 20 different sites, upstream and downstream the selected WWTPs, in two different seasons. Analysis was performed by solid phase extraction followed by liquid chromatography coupled to tandem mass spectroscopy. Pharmaceuticals were detected in 27.8% of the samples. Selective serotonin reuptake inhibitors (SSRIs), anti-inflammatories and antibiotics presented the highest detection frequencies (27.8, 23.6 and 23.6%, respectively) and average concentrations (37.9, 36.1 and 33.5 ng L − 1 , respectively). When assessing the impact of WWTPs, an increase of 21.4% in the average concentrations was observed in the samples located downstream these facilities, when compared with the upstream samples. Increased detection frequencies and concentrations were observed at lower flow rates, both when comparing summer and winter campaigns and by evaluating the different rivers. Risk quotients (RQs) higher than one were found for two pharmaceuticals, concerning two trophic levels. However, since Iberian rivers are highly influenced by water scarcity, in drought periods, the flow rates in these rivers can decrease at least ten times from the lowest value observed in the sampling campaigns. In these conditions, RQs higher than 1 would be observed for 5 pharmaceuticals, additionally, all the detected pharmaceuticals (11) would present RQs higher than 0.1. These results emphasize that the river flow rate represents an important parameter influencing pharmaceuticals concentrations, highlighting the ecotoxicological pressure, especially due to water scarcity in drought periods. This should be a priority issue in the environmental policies for minimizing its impact in the aquatic environment.

Ochratoxin A (OTA) is mainly found in cereals and cereal-based foodstuffs, but also in wine. Being one of the most consumed alcoholic drinks in Portugal and one of the main sources of human exposure to OTA, wine monitoring and exposure studies are essential. The analytical methodology consisted of the direct injection of the filtered samples into the liquid chromatograph, equipped with fluorescent detection (LC-FLD). Linearity was adequate, both in mobile phase and in matrix-matched solutions, with R

In line with the Directive 2013/39/EU the most representative surface waters, regarding pharmaceuticals contamination, were selected based on a Portuguese nationwide monitoring exercise. To meet this purpose, and given that wastewater treatment plants (WWTPs) are regarded as the major point sources of pharmaceuticals environmental contamination, the occurrence, fate and environmental risk assessment (ERA) of eleven of the most consumed pharmaceuticals, belonging to several therapeutic classes were assessed in 15 WWTPs (influents (WWIs) and effluents (WWEs)), from five different regions during one year (4 sampling campaigns). Results showed that all samples were contaminated with at least 1, and up to 8 from the 11 targeted pharmaceuticals. The highest concentrations observed were 150 and 33 μg L(-1) for WWI and WWE, respectively. Regarding temporal and spacial influence, winter, Alentejo, Algarve and Center regions presented higher mass loads. The ERA posed by 7 of the selected pharmaceuticals presented a risk quotient higher than 1 to the three trophic levels. Our findings highlighted that the rivers Mondego, Tagus, Ave, Trancão, Fervença and Xarrama should be selected as surface water monitoring stations. This study gives a good overview on pharmaceuticals contamination in WWTPs and its impact on surface waters in Portugal. Thus, a more integrative approach to rank and prioritize pharmaceuticals, based on an integrated assessment of ERA and exposure of surface water, was provided to support the future selection of the 6 most representative monitoring stations in Portugal, as required by the above mentioned directive.

Consumption of tea and medicinal plant infusions in general has been increasing. The worldwide growing consumption rate of tea and infusions is associated to the health promoting properties and a healthy lifestyle trend, and also to the assumption of minimal or no toxic side effects when consumed in large quantities compared with synthetic drugs. However, tea and the herbs used in the preparation of infusions can be contaminated with mycotoxins, for which no maximum levels are currently established. In this context this study aimed to analyze commercially available teas and medicinal plants destined to be consumed as infusions and assess the resulting exposure to the mycotoxins AFs and ZEA. With such purpose, 37 (convenience) samples of tea and medicinal plants destined to be consumed as infusions were analyzed through a SPE-ELISA analytical set-up. Nearly 84% of the analyzed samples were positive for at least one of the mycotoxins and 43.24% were positive for both AFs and ZEA. None of the samples negative for both AFs and ZEA were tea (i.e. derived from Camellia sinensis). Approximately 65% of the analyzed samples were positive for AF (14.74 ± 7.18 μg/kg). In general, teas featured the highest AFs contamination levels versus medicinal plants. Is it also noteworthy that all analyzed tea samples were AF-positive. Sixty two percent of the samples were contaminated with a ZEA (8.87 ± 5.31 μg/kg). Statistically significantly higher levels of ZEA in organically produced samples were observed. Samples sold in bulk (i.e. not packed) were those with the highest ZEA levels. The highest maximum levels of AFs and ZEA were detected in chamomile flowers (28.15 μg/kg), the second most consumed medicinal plant in Portugal. The degree of exposure of the Portuguese to these mycotoxins through consumption of tea and medicinal plants destined to prepare infusions, was assessed through the Estimated Daily Intake (EDI). For AFs the EDI was 0.13 ng/kg bw/day, which represents 64.12% of the proposed TDI. In the risk assessment, the worst scenario corresponded to conventional produced medicinal plants (122.45%). Conversely, tea and medicinal plants contribute little to the exposure of the Portuguese population to ZEA (EDI = 0.08 ng/kg bw/day). The obtained results proved the need for further studies to evaluate the occurrence of these mycotoxins in these food commodities of increasing consumption, in order to monitor the exposure of the population and, if justified, to serve as the basis for establishing maximum limits for the control of exposure.

A study was performed to evaluate the risk regarding the presence of nickel in commercial premade baby foods: 26 ready meals, 31 fruits, 8 deserts and 20 paps. The analytical methodology used for the determination of nickel was dry ashing followed by graphite furnace atomic absorption spectrometry. Method detection limit (12.5 μg kg−1) was obtained through a matrix matched calibration curve, using a nickel free commercial ready meal (meat). The results showed that 78 (91.8 %) samples contained nickel, with values up to 225.7 μg kg−1. The group with the highest average concentration was the fruits (50.1 μg kg−1) followed by the ready meals (40.2 μg kg−1). Samples from organic farming presented higher detection frequency and average concentration (54.7 μg kg−1) than the others. Using the average results, the tolerable daily intake proposed by EFSA (2.8 μg kg−1 b.w.) was surpassed for the 2 year old age group, with daily ingestions up to 3.1 μg kg−1 b.w.. This results highlight the risk promoted by the nickel present in commercial premade baby food.

Until now, the available data regarding citrinin (CIT) levels in food and the consumption of contaminated foods are insufficient to allow a reliable estimate of intake. Therefore, biomonitoring configuring analysis of parent compound and/or metabolites in biological fluids, such as urine or blood, is being increasingly applied in the assessment of human exposure to CIT and its metabolite, dihydrocitrinone (DH-CIT). Most studies report urinary levels lower for the parent compound when compared with DH-CIT. A high variability either in the mean levels or in the inter-individual ratios of CIT/DH-CIT between the reported studies has been found. Levels of DH-CIT in urine were reported as being comprised between three to seventeen times higher than the parent mycotoxin. In order to comply with this objective, sensitive analytical methodologies for determining biomarkers of exposure are required. Recent development of powerful analytical techniques, namely liquid chromatography coupled to mass spectrometry (LC-MS/MS) and ultra-high-performance liquid chromatography (UHPLC-MS/MS) have facilitated biomonitoring studies, mainly in urine samples. In the present work, evidence on human exposure to CIT through its occurrence and its metabolite, in biological fluids, urine and blood/plasma, in different countries, is reviewed. The analytical methodologies usually employed to evaluate trace quantities of these two molecules, are also presented. In this sense, relevant data on sampling (size and pre-treatment), extraction, cleanup and detection and quantification techniques and respective chromatographic conditions, as well as the analytical performance, are evidenced.

Potential risks associated with releases of human pharmaceuticals into the environment have become an increasingly important issue in environmental health. This concern has been driven by the widespread detection of pharmaceuticals in all aquatic compartments. Therefore, 22 pharmaceuticals, 6 metabolites and transformation products, belonging to 7 therapeutic groups, were selected to perform a review on their toxicity and environmental risk assessment (ERA) in different aquatic compartments, important issues to tackle the water framework directive (WFD). The toxicity data collected reported, with the exception of anxiolytics, at least one toxicity value for concentrations below 1 µg L−1. The results obtained for the ERA revealed risk quotients (RQs) higher than 1 in all the aquatic bodies and for the three trophic levels, algae, invertebrates and fish, posing ecotoxicological pressure in all of these compartments. The therapeutic groups with higher RQs were hormones, antiepileptics, anti-inflammatories and antibiotics. Unsurprisingly, RQs values were highest in wastewaters, however, less contaminated water bodies such as groundwaters still presented maximum values up to 91,150 regarding 17α-ethinylestradiol in fish. Overall, these results present an important input for setting prioritizing measures and sustainable strategies, minimizing their impact in the aquatic environment.

Potential risks associated with releases of human pharmaceuticals into the environment have become an increasingly important issue in environmental health. This concern has been driven by the widespread detection of pharmaceuticals in all aquatic compartments. Therefore, 22 pharmaceuticals, 6 metabolites and transformation products, belonging to 7 therapeutic groups, were selected to perform a systematic review on their source, fate and occurrence in different aquatic compartments, important issues to tackle the Water Framework Directive (WFD). The results obtained evidence that concentrations of pharmaceuticals are present, in decreasing order, in wastewater influents (WWIs), wastewater effluents (WWEs) and surface waters, with values up to 14 mg L−1 for ibuprofen in WWIs. The therapeutic groups which presented higher detection frequencies and concentrations were anti-inflammatories, antiepileptics, antibiotics and lipid regulators. These results present a broad and specialized background, enabling a complete overview on the occurrence of pharmaceuticals in the aquatic compartments.

Ochratoxin A (OTA) is a mycotoxin detected worldwide. Urine biomonitoring is a more realistic and non-invasive way to assess exposure when compared to traditional methods based on food occurrence and consumption data. Few studies have investigated children's exposure to OTA, although it is a more susceptible population. Our main goal was the OTA biomonitoring in urine of Portuguese children to better characterize the health risk of this population. The validated analytical methodology was based on an immunoaffinity clean-up, followed by LC-FD. First morning samples were collected, between 2018 and 2019, from 85 healthy children, 41 boys and 44 girls, aging between 2 and 13 years old. Overall, from the 85 analysed samples 79 (92.94%) were found positive. The average OTA concentration level was of 0.020 ng/mL (29.41 ng/g of creatinine), with a maximum value of 0.052 ng/mL (114.45 ng/g of creatinine). No statistical differences were found between the OTA contamination levels, the anthropometric data and dietary habits considered. Depending on the excretion rate considered for the PDI calculation, the risk assessed for the average OTA contamination varied from 10% to 194%, indicating a potential concern. This is the first study regarding OTA exposure in Portuguese children, showing their clear exposure.

Antibiotics, especially fluoroquinolones (FQs), have been largely used in animal husbandry namely poultry production. Therefore, this study aimed to identify, quantify and estimate the daily intake for adult and 3-year-old populations of the FQs norfloxacin (NOR), ciprofloxacin (CIP) and enrofloxacin (ENR) through poultry muscle consumption. The results showed detection frequencies of 78% and 62% in supermarket and school canteen samples, respectively. Of the 182 analysed samples, 4 did not comply with ENR maximum residue level (MRL), and 9 were contaminated with NOR, not allowed as a veterinary medicine of food-producing animals. The highest estimated daily intake value was obtained for the 3-year-old population regarding the sum of ENR and CIP (0.46 μg kg−1 day−1); value substantially lower than the established acceptable daily intake (2.0 μg kg−1 day−1). Although the low risk found, the high detection frequencies support the apprehension of the different international organizations, towards the emergence of human bacterial resistances to FQs originated from poultry production.

The occurrence of zearalenone (ZEA) in maize bread, broa, for human consumption, from the Portuguese market, was evaluated. Good analytical performance was obtained through extraction with acetonitrile:water (90:10), clean-up with immunoaffinity columns, and detection and quantification by liquid chromatograph with-tandem mass spectrometry. ZEA levels were determined in 52 samples to verify the compliance with the maximum permitted levels by the European legislation. One broa sample exceeded the maximum limit established. A higher contamination frequency was observed in samples in which maize and wheat were present. Overall, 13.5% of the samples were contaminated, at levels oscillating between 9.6 and 50.4 μg/kg. Considering the percentage of the tolerable daily intake (TDI) obtained, 0.6%, the risk assessment linked with the exposure to ZEA was considered very low for the studied population. This is the first study on the intake assessment of ZEA through maize bread consumption in Portugal.

Several toxic effects have been described after exposure to ochratoxin A (OTA), which can enter the human diet directly through food or through animal products via carry-over from contaminated feed. To assess the exposure of the Portuguese population to OTA, a study over a two-year period was conducted. It involved analysis of 472 morning urine samples from inhabitants of four regions, together with a survey of regional bread (738) and pork (254) samples. These foodstuffs are two staple foods in the Portuguese and Mediterranean diet that present a high and widespread consumption by the majority of the population. The bread samples analysed showed a low level of contamination, although the contamination range was broad and some of the samples exceeded the maximum level established in the European Union. Maize bread (broa), especially when made by mixing with rye, was the most contaminated, followed by whole grain-, rye- and wheat-based bread. However, the latter contributed more to OTA exposure, because they were more commonly consumed. Even though the occurrence was rather low, the average OTA amount in pork was relatively high compared to previous national and other surveys. The observed high within-subject variability of OTA in urine limited the use of this biomarker of exposure at the individual level, but not on a population or subgroup of subjects scale. Among the studied population, a widespread exposure was confirmed by the high frequency of OTA contamination in urine, although characterised by a low average contamination level. Independent of region or population, pork appears to be the main contributor to the daily OTA intake in Portugal. These data were clearly in contrast with previous studies showing that cereals and their derived products were the major contributors, while food of animal origin only contributed a small part to the total human dietary OTA exposure.

This study provides the first environmental risk assessment (ERA) for zearalenone (ZEN), the only known mycoestrogen, based on a broad-scale investigation on its occurrence in rivers and creeks from Portugal. Water sample filtration and immunoaffinity columns (IAC) clean-up followed by liquid chromatograph with tandem mass spectrometry (LC/MS n ) provided an analytical method with good analytical performance. ZEN levels were determined for seven Portuguese rivers and one creek, during two different seasons, in a total of 38 samples collected upstream wastewater treatment plants (WWTPs). Overall, 23.7% were contaminated with ZEN at levels ranging between 5.6 and 82.6 ng/L. The highest concentration was observed during spring, although no statistically significant differences were observed between spring and autumn sampling campaigns. The potential ecotoxicological risk from ZEN to different trophic levels of aquatic organisms was evaluated by means of risk quotients (RQs) calculation. Although all the RQs obtained were lower than 1, our results confirm that ZEN is a relatively frequent contaminant in flowing waters in Portugal and might contribute to the overall estrogenic activity in the environment.

A critical evaluation of the European Medicines Agency (EMA) Guideline on Environmental Risk Assessment (ERA) was performed on 16 of Portugal's most consumed pharmaceuticals in wastewater effluents (WWEs), the main route for aquatic contamination. The predicted environmental concentrations (PECs) were formulated based on the Guideline, after incorporating several refinements. The best approach was selected by comparing the measured environmental concentrations (MECs) to the PECs in WWEs. Finally, risk was assessed by comparing PECs to predicted no-effect concentrations (PNECs). The results showed that the default value of the penetration factor (Fpen) used by the EMA (0.01) was surpassed and that national consumption and excretion data were the two most important parameters for PEC calculations. The risk quotient between PECs and PNECs was higher than 1 for 12 pharmaceuticals, indicating a risk to all three trophic levels of aquatic organisms (algae, daphnids and fish). To improve the current ERA framework, suggestions were made for incorporating consumption and excretion data, changing the default value of Fpen to 0.04 and adding a safety factor of 10. Moreover, this evaluation should be performed for pharmaceuticals already on the market, and future ERAs should incorporate a risk-benefit analysis, an important risk-management step.

The occurrence, fate, seasonal influence and environmental risk assessment of four selective serotonin re-uptake inhibitors (SSRIs) antidepressants, citalopram, fluoxetine, paroxetine and sertraline, were studied in 15 different wastewater treatment plants (WWTPs) across Portugal. Influent and effluent samples from four sampling campaigns, in 2013, were extracted through Oasis HLB cartridges, and quantified through liquid chromatography with tandem mass spectrometry (LC-MSn).Results showed that citalopram was the SSRI most frequently found, both in influents and in effluents, with mean mass loads ranging between 14.56 and 9.51 mg/day/1000 inhabitants, respectively. Fluoxetine and sertraline were only detected in influent samples, in lower mean mass loads (14.60 and 1.36 mg/day/1000 inhab., respectively), whereas paroxetine was found in influent and effluent samples (12.61 and 18.90 mg/day/1000 inhab., respectively). WWTPs were not capable of completely removing these pharmaceuticals; nonetheless, the mean removal efficiency was 82.24%. Removal efficiency was lower in winter (74.21%), summer (72.02%), and autumn (81.19%), when compared to spring (100%). Our results translate the variations in SSRI prescription and use between the five Portuguese regions in study. Influent contaminated samples were found in WWTPs from Lisbon, Alentejo, Center and North (28.25, 19.01, 16.55 and 6.98 mg/day/1000 inhab., respectively). In the Algarve region no contaminated samples were found. A seasonal pattern in the presence of SSRIs in influent wastewaters was observed. The SSRIs mass loads in influent wastewaters were higher in autumn, followed by spring, winter, and summer. Finally, the potential ecotoxicological risk posed by SSRIs to different trophic levels of aquatic organisms, exposed to the effluent wastewaters studied was evaluated by means of risk quotients (RQ). Citalopram and paroxetine, the only SSRIs found in these samples, presented RQ lower than 1. According to the results, algae appeared to be the most sensitive followed by fish and daphnids.

A new analytical methodology, based on liquid chromatography with fluorescence detection (LC-FD), after extraction, enzymatic hydrolysis, and solid-phase extraction (SPE) through Oasis HLB cartridges, was developed and validated for the simultaneous determination of three monohydroxy derivatives of polycyclic aromatic hydrocarbons (PAHs). The optimized analytical method is sensitive, accurate, and precise, with recoveries between 62 and 110% and limits of detection of 227, 9, and 45 ng/g for 1-hydroxynaphthalene, 2-hydroxyfluorene, and 1-hydroxypyrene, respectively. Their levels were estimated in different cephalopod matrices (edible tissues and hemolymph). The methodology was applied to samples of the major cephalopod species consumed worldwide. Of the 18 samples analyzed, 39% were contaminated with 1-hydroxynaphthalene, which was the only PAH metabolite detected. Its concentration ranged from 786 to 1145 ng/g. This highly sensitive and specific method allows the identification and quantitation of PAH metabolites in forthcoming food safety and environmental monitoring programs.

Selective serotonin reuptake inhibitor (SSRI) antidepressants are among the most prescribed pharmaceuticals worldwide and are ubiquitous in different areas of the environment. There are few reliable and specific analytical methods for the detection and quantification of SSRIs in environmental samples. The majority of the reported methods generally involve cleanup procedures that usually employ solid-phase extraction (SPE) followed by liquid chromatography with tandem mass spectrometry detection (LC-MS-MS). Herewith, a comprehensive overview of the described analytical methods available for the determination of SSRIs in environmental samples is provided. We address all steps involved in the analytical procedures, with the inherent advantages and disadvantages of each discussed.

A reliable exposure assessment of mycotoxin contamination relies on their unambiguous identification and accurate quantification. With such purpose, a new analytical methodology was developed for evaluation of ochratoxin A (OTA) contamination in pork. Briefly OTA extraction from minced muscle samples involved the use of acidified methanol, ultrasound treatment and centrifugation, followed by an immunoaffinity column (IAC) clean-up step before mass spectrometric detection (precursor-to-fragment transitions: m / z 404 → m / z 358 and m / z 404 → m / z 386) in positive ESI mode using SRM scanning. The method combines green chemistry principles (e.g. absence of highly toxic solvents commonly used in this matrix) with a validation according to the guidelines laid down by the 2002/657/EC European Decision parameters: recoveries varied between 98.5% and 100.6%, limits of detection (LOD) and quantification (LOQ) were estimated at 0.06 and 0.19 μg/kg, whereas decision limit (CCα) and detection capability (CCβ) were 0.01 and 0.50 μg/kg, respectively. The proposed analytical set-up was successfully applied to twenty pork samples commercially acquired.

Selective serotonin re-uptake inhibitors (SSRIs) antidepressants are among the most prescribed pharmaceutical active substances throughout the world. The occurrence of these widely used compounds in different environmental compartments (wastewaters, surface, ground and drinking waters, and sediments), justify the growing concern about these emerging environmental pollutants. Viewing an ecopharmacovigilance approach, a comprehensive discussion of the state of the art regarding different contamination sources, fate, degradation and occurrence is presented. Information on the current distribution levels and fate in different environmental matrices continues to be sparse and measures are imperative to improve awareness and encourage precautionary actions to minimize SSRIs' environmental impact.

Ochratoxin A (OTA) is an important mycotoxin with potential to reach the human food chain through carry-over of contaminated, mostly cereal-based, feed into animal-derived products. Certain population groups, such as infants and children, are intensive and relatively restricted consumers of some animal-derived products, particularly milk and other dairy products, which may become contaminated with OTA. This review examines the literature on the occurrence of OTA in animal-derived products and discusses the public health and food safety implications of consumption of these products. The risk of OTA contamination of meat, milk, blood and derived products is discussed.

Due to strong implications for food safety, control of fluoroquinolones residues in swine meat should be undertaken to verify compliance of the contamination levels with the maximum residue limits recently updated by Commission Regulation (EU) No. 37/2010 of 22 December 2009. Solid-phase extraction is widely used in antibiotic analysis in food of animal origin. In this study, the results of a comparative study using different types of solid-phase extraction columns, HLB, MCX, and MAX, for ciprofloxacin and enrofloxacin analysis, in pork meat, are presented. In addition, diverse sample treatments for defatting, precipitate proteins, eliminate cations, and increase the ionic strength, were used to obtain the most suitable method of analysis. Only the MCX's use followed by liquid chromatography with fluorescence detection resulted in chromatograms that allow the quantification at maximum residue limits. The validation method, in terms of CCα and CCβ, recovery and precision determination, was according to the EU Decision 2002/657/EC. This procedure was used in the analysis of 50 samples of pork meat of Portuguese origin. Only two samples presented residues of enrofloxacin at 30 and 42 μg/kg, values under the legal maximum residue limit.

Ochratoxin A (OTA) is a nephrotoxic mycotoxin that has received particular attention because of the toxic effects, widespread occurrence in contaminated food and feed chain, suspected causal effect on nephropathies, and, more recently, possibility of exposure by inhalation in domicile and occupational settings. Biomarkers have been used not only to ascertain the role of OTA in inducing chronic renal failure diseases, but also as a means to portray general populations' risk to the mycotoxin. Biomonitoring can thus be used to assess internal OTA exposure, with no need to recognize the main source of exposure. And so it presents undeniable advantages over the monitoring of external dose. With a just right understanding of biomarkers, it is possible to follow the trail from exposure right to effect, and so contribute both to surveillance plans and etiological studies. In recognition of the long serum half-life and the renal elimination of OTA, most of the studies present serum/plasma and/or urine analyses as markers of exposure. In this review and for each of these main matrices, a comparison over the advantages and disadvantages is offered. Although currently limited, an overview of the current knowledge on OTA biomarkers and the influential role of the individual characteristics, namely gender and age, along with season and geographical location is given. Attention is also given to the ongoing debate over the existence of OTA-DNA adducts, a biomarker of effective dose regarded as an alternative to biomarkers of internal dose. Although unspecific, OTA effect biomarkers are also reviewed.

Wine and grape juices were identified as one of the most important sources of ochratoxin A (OTA), a mycotoxin with diverse toxic effects that naturally appears in food and foodstuffs all over the world. The aim of this study was to assess the OTA levels in Portuguese wines through the application of a simple and accurate method based on liquid chromatography (LC) with direct injection, followed by fluorescence detection (FD). Randomly selected wine samples were used to evaluate the performance of direct injection as efficient, fast, inexpensive and safe sample preparation method. The proposed method was successfully validated. The limit of quantification (LOQ) was 1.0 microg/L and OTA recoveries from wine samples, spiked at the three fortification levels, were higher than 85.4%, with RSDs lower than 9.6% for both red and white wines. The presence of OTA was confirmed by methyl ester derivatization followed by LC analysis. Data on OTA levels were obtained for 60 Portuguese red and white wine samples. OTA was found in 12 samples, nine (26%) red wine samples and three (12%) white wine samples. Only one red wine sample and one white wine sample presented a contamination level above the LOQ, with 1.23 and 2.4 microg/L, respectively. It should be pointed out that this white wine sample exceeded the EC maximum permitted level of 2.0 microg/L. The safe dose established as 120 ng/kg body weight/week was not exceeded by the weekly intake estimated for the samples contaminated above the LOQ.

An analytical strategy, for the determination of tetracyclines (TCs), based on a HPLC system coupled with a photo-reactor followed by post-column derivatization was developed. Higher fluorescence emission after coupling the resulting photo-fragments with magnesium ions was observed for the determination of minocycline (MC), epitetracycline (ETC), tetracycline (TC) and doxycycline (DC). The manifold included a HPLC system with a photo-reactor (PTFE tubing helically coiled around a low-pressure mercury lamp), a mixing T-piece and a fluorescence detector. The derivatization reagent was delivered at 0.5 mL min −1 by a pump. After HPLC separation using a gradient system with a mobile phase containing oxalic acid 0.02 M and acetonitrile, TCs were irradiated for 60 s, and the resulting photo-fragments were mixed with the post-column derivatization reagent, and the magnesium derivatives of TCs were detected by fluorimetry ( λ exc 386 nm, λ em 500 nm). The results obtained showed a significant increase of sensitivity due to photodegration of TCs, 45.4%, 37.6% and 25.3% for MC, TC and ETC respectively. For DC an increase of only 1.5% was observed. The developed method was successfully applied to TCs determination in hospital and municipal wastewater samples using solid phase extraction with Oasis HLB cartridges. The LOQs were 0.25, 0.15, 01 and 0.25 μg L −1 for TC, ETC, MC and DC, respectively. The recovery values oscillated between 107.1% and 92.4% for fortification of 2.5 μg L −1 of each antibiotic.

A reliable method for the determination of benzoic and sorbic acids, caffeine and saccharin in soft drink and nectars using high performance liquid chromatography and UV detection was validated. The chromatographic separation was achieved with a C18 column (250 × 4.6 mm) and one buffered mobile phase, KH2PO4 0.02 M/ACN (90:10)/phosphoric acid at pH 4.2. The effluent was monitored at 220 nm. Benzoic acid was detected in 19 soft drinks, being 11 traditional soft drinks and 8 based on mineral water, with mean concentrations of 158 and 148 mg/L, respectively. Fifteen samples, 7 traditional and 8 based on mineral water, contained sorbic acid, with mean concentrations of 172 and 188 mg/L, respectively. Saccharin and caffeine were not detected in soft drinks based on mineral waters. In contrast, 6 samples of traditional drinks contained both additives, with mean levels of 75 and 97 mg/L, respectively. The four additives were not detected in nectar samples. According to European Union and Portuguese legislation, the maximum permitted level (MPL) for benzoic acid was exceeded in 10 soft drinks, and the MPL for sorbic acid overlapped in 3 samples of the referred samples. Two samples exceeded the MPL for the sum of both preservatives, and in 1 sample the MPL for saccharin was overlapped. The estimated daily intakes (EDI) of benzoic, sorbic acid, and saccharin for the average consumer were below the acceptable daily intakes (ADIs). For benzoic acid, the EDIs were 0.25 and 0.32 mg/kg b.w./day, representing 4.9%, and 6.4% of the ADI, respectively for traditional soft drinks and soft drinks based on mineral water. A similar situation was observed for sorbic acid. In this way, the EDIs were 0.17 and 0.41 mg/kg b.w./day, representing only 0.68% and 1.6% of the ADI for the same kind of drinks. For saccharin, the EDI represents 1.28% of the ADI. The EDI for caffeine was 0.08 mg/kg b.w./day.

The occurrence of OTA in fresh and packed wheat and in maize bread and the evaluation of the exposure degree through their consumption in two Portuguese populations from Porto and Coimbra, during the winter of 2007, were studied. One hundred and sixty eight bread samples, 61 maize and 107 wheat, were analysed by liquid chromatography–fluorescence detection (LC–FD). The results showed that 84% of samples were contaminated, with a maximum level of 3.85 ng/g (above the EU maximum limit, 3 ng/g). Fresh wheat bread presented higher levels than packed wheat bread. Moreover, the traditional maize bread, in either city, was consistently more contaminated than wheat bread, 0.25 vs 0.19 ng/g, and 0.48 vs 0.34 ng/g for Porto and Coimbra, respectively. Avintes maize bread showed the highest mean contamination and maximum levels. The higher estimated daily intake of OTA from both types of bread in the population of Coimbra compared to Porto reflects the higher average contamination of bread in the first city.

This study investigated the occurrence of tetracyclines (TCs), namely minocycline (MIN), TC, and its epimer epitetracycline (ETC), and doxycycline (DC), in four hospital wastewater effluents and its fate in municipal wastewater treatment plants (WWTPs), in Coimbra, Portugal. Analytical determination was carried out by solid-phase extraction followed by liquid chromatography with fluorescence detection. A gradient system with a mobile phase containing oxalic acid 0.02 M and acetonitrile was used. After postcolumn derivatization with magnesium reagent, TCs were detected at lambda(exc) 386 nm and lambda(em) 500 nm. The proposed method allowed good sensitivity, accuracy, and precision. LOQs were 0.5 microg l(-1) for ETC and TC and 15 and 5 microg l(-1) for MIN and DC, respectively. The recovery values ranged between 66.4% and 117.1%, and intraday and interday repeatability was lower than 6.8%. The method was successfully used to determine the presence of the above-mentioned TCs in 24 wastewater composite samples obtained from hospital effluents and from influent and effluent of the WWTP located in Coimbra, Portugal. MIN and TC were found in 41.7% of the samples; ETC and DC were found in 25% and 8.3% of the samples, respectively. The levels found ranged from 6 to 531.7 microg l(-1) in hospital effluents, while its concentrations in WWTP ranged from 95.8 to 915.3 microg l(-1). A seasonal influence in the concentrations found has also been observed, the levels found in samples collected during spring being higher than those observed in samples collected during autumn; however, these are only preliminary results. The WWTP removal rate ranged between 89.5% and 100%.

To trace the contamination and evaluate the environmental impact of piggeries, the occurrence of norfloxacin (NOR), ciprofloxacin (CIPRO), enrofloxacin (ENRO) and sarafloxacin (SARA) residues were evaluated in water for swine consumption, piggeries' wastewaters, drainage waters and a river receiving piggery effluents. Water samples were acidified before being percolated through Oasis HLB cartridges or through a tandem system consisting of strong anion-exchange and Oasis HLB cartridges. A liquid chromatography with fluorescence detection (LC-FD) method developed in previous studies, based on the use of a monolithic column, was successfully applied. NOR, CIPRO, ENRO and SARA had good linear responses, with mean regression coefficients higher than 0.9989. The limits of quantification (LOQ) for waters for swine consumption, and a river receiving piggery effluents, were 0.025 ug L(-1) for NOR, CIPRO and ENRO and 0.05 ug L(-1) for SARA. For wastewaters and drainage waters, the LOQs were 0.05 ug L(-1) for CIPRO and ENRO and 0.1 ug L(-1) for SARA. Recoveries obtained for spiked samples ranged from 70.8 to 89.1%, and precision data were under 12.7%. The method was successfully applied to 18 environmental water samples, collected from piggeries in Alentejo, Portugal, and the first Portuguese data are presented. ENRO was detected in a total of 8 samples, at concentrations ranging between 0.31 microg L(-1) and 63 microg L(-1). CIPRO, its degradation product, and SARA were present in 2 samples, in the ranges 0.48-0.83 microg L(-1) and 1.8-14 microg L(-1), respectively. No fluoroquinolones were detected in the waters for swine consumption or in the river water samples.

Ochratoxin A (OTA) is one of the most studied mycotoxins, with great public health and agroeconomic significance. The exposure of both humans and animals to this fungal toxin has been associated to food matrices since its discovery. However, according to recent reports, OTA may also represent a potential airborne hazard, in water-damaged buildings, or occupational contamination, in workplaces with high mould exposure, such as agricultural, farm and alimentary industries. Further, in addition to the conventional studied food matrices, worldwide consumed, there are increasing reports on different and less obvious sources of alimentary exposure. These include traditional and home-made food products, highly consumed by specific groups of populations that thus might be at risk. This paper aims to spotlight the current knowledge on these non-conventional OTA exposure sources.

The presented procedure involves an extraction with methanol–water, centrifugation and cleanup with immunoaffinity columns. A comparison study between fluorescence detector, mass spectrometry, and tandem mass spectrometry with a triple quadrupole (QqQ) analyzer using an electrospray ionisation interface for the determination of fumonisin B1 and B2 in corn-based products has been performed. Limits of quantification obtained by the three detectors were lower than the maximum levels established by European Commission. Liquid chromatography coupled to tandem mass spectrometry provides higher sensitivity (12 μg kg−1for fumonisins B1 and B2) when compared to mass spectrometry (40 μg kg−1for both fumonisins), and fluorescence detection (20 μg kg−1 for fumonisin B1 and 15 μg kg−1 for B2), and also showed to be more precise. At 150 and 250 μg kg−1 spiking levels, the recovery rates for fumonisin B1 and B2 in corn products varied from 79% to 102%, with a relative standard deviation ranging from 9% to 17%. A critical assessment including advantages and drawbacks of each technique is presented. A total of 41 organic and non-organic corn-based food samples from Valencia markets were analyzed. Seven samples were contaminated with levels ranging from 68 μg kg−1 to 922 μg kg−1of fumonisin B1 and 42 μg kg−1 to 640 μg kg−1of fumonisin B2. Only one sample exceeded the maximum level for the sum of fumonisin B1 and B2, proposed for corn products in a recent EU regulation. The contamination frequency of organic corn samples (40%) was higher than non-organic ones (3.7%), and contained higher levels of fumonisin B1 and B2.

In a survey of levels of acesulfame-K and aspartame in soft drinks and in light nectars, the intake of these intense sweeteners was estimated for a group of teenage students. Acesulfame-K was detected in 72% of the soft drinks, with a mean concentration of 72 mg l(-1) and aspartame was found in 92% of the samples with a mean concentration of 89 mg l(-1). When data on the content of these sweeteners in soft drinks were analysed according to flavour, cola drinks had the highest mean levels for both sweeteners with 98 and 103 mg l(-1) for acesulfame-K and aspartame, respectively. For soft drinks based on mineral water, aspartame was found in 62% of the samples, with a mean concentration of 82 mg l(-1) and acesulfame-K was found in 77%, with a mean level of 48 mg l(-1). All samples of nectars contained acesulfame-K, with a mean concentration of 128 mg l(-1) and aspartame was detected in 80% of the samples with a mean concentration of 73 mg l(-1). A frequency questionnaire, designed to identify adolescents having high consumption of these drinks, was completed by a randomly selected sample of teenagers (n = 65) living in the city of Coimbra, in 2007. The estimated daily intakes (EDI) of acesulfame-K and aspartame for the average consumer were below the acceptable daily intakes (ADIs). For acesulfame-K, the EDI was 0.7 mg kg(-1) bw day(-1) for soft drinks, 0.2 mg kg(-1) bw day(-1) for soft drinks based on mineral waters, and 0.5 mg kg(-1) bw day(-1) for nectars, representing 8.0%, 2.2%, and 5.8% of the ADI, respectively. A similar situation was observed for aspartame. In this way, the EDI for soft drinks was 1.1 mg kg(-1) day(-1), representing only 2.9% of the ADI. In respect of nectars, the EDI was 0.2 mg kg(-1) bw day(-1), representing 0.5% of the ADI. Soft drinks based on mineral waters showed the lowest EDI values of 0.3 mg kg(-1) bw day(-1), accounting for 0.7% of the ADI.

Fluoxetine, a selective serotonin re-uptake inhibitor (SSRI) antidepressant, is among the most prescribed pharmaceutical active substances worldwide. This study aimed to assess its accumulation and metabolization in the mussel Mytillus galloprovincialis, considered an excellent sentinel species for traditional and emerging pollutants. Mussels were collected from Ria Formosa Lagoon, Portugal, and exposed to a nominal concentration of fluoxetine (75 ng L-1) for 15 days. Approximately 1 g of whole mussel soft tissues was extracted with acetonitrile:formic acid, loaded into an Oasis MCX cartridge, and fluoxetine analysed by liquid chromatography with tandem mass spectrometry (LC-MSn). After 3 days of exposure, fluoxetine was accumulated in 70% of the samples, with a mean of 2.53 ng g(-1) dry weight (d.w.) and norfluoxetine was only detected in one sample (10%), at 3.06 ng g(-1) d.w. After 7 days of exposure, the accumulation of fluoxetine and norfluoxetine increased up to 80 and 50% respectively, and their mean accumulated levels in mussel tissues were up to 4.43 and 2.85 ng g(-1) d.w., respectively. By the end of the exposure period (15 days), both compounds were detected in 100% of the samples (mean of 9.31 and 11.65 ng g(-1) d.w., respectively). Statistical analysis revealed significant accumulation differences between the 3rd and 15th day of exposure for fluoxetine, and between the 3rd and 7th against the 15th day of exposure for norfluoxetine. These results suggest that the fluoxetine accumulated in mussel tissues is likely to be metabolised into norfluoxetine with the increase of the time of exposure, giving evidence that at these realistic environmental concentrations, toxic effects of fluoxetine in mussel tissues may occur. (C) 2016 Elsevier Ltd. All rights reserved.

An analytical study based on extraction with acetonitrile-water, immunoaffinity column cleanup, and HPLC/fluorescence detection for separation and identification of ochratoxin A in coriander was carried out. Validation of the applied methodology was done through accuracy and precision studies. Homogenized samples of coriander were spiked in triplicate with ochratoxin A at 0.5, 1.0, 2.0, and 5.0 µg/kg levels. Recovery values were in the range of 98% for a fortification level at 0.5 µg/kg to 109.1% at 1.0 µg/kg. Application to coriander samples available in Portuguese markes showed no contamination with ochratoxin A.

Ochratoxin A (OTA) is a secondary fungal metabolite produced by several moulds, mainly by Aspergillus ochraceus and by Penicillium verrucosum, that occurs in meat products. The aim of this work was to optimize an efficient extraction procedure for the determination of OTA in muscle tissue in order to assess its occurrence in meat samples. Three different apparatus, a Waring blender, a switching apparatus, and an ultrasonic processor, were evaluated to verify the efficiency of extraction. The analytical methods proposed involve the extraction with chloroform-orthophosphoric acid, cleanup through an immunoaffinity column, high-performance liquid chromatography/fluorescence detection for separation and identification of OTA, and confirmation with liquid chromatography/FD after methylation of OTA in muscle tissue. The limit of quantification of the proposed method was 0.04 microg kg(-1). Recoveries of OTA, using switching apparatus, ranged from 90.3 to 103.2% for chicken muscle spiked at 2.4 and 0.48 microg kg(-1), respectively, with a within-day relative standard deviation of 17 and 15.3%. The proposed method was applied to 38 chicken, swine, and turkey muscle samples and the presence of OTA was confirmed in five samples. Finally, the estimated daily intake of OTA in this study was between 23 pg kg(-1) body weight per day for swine samples and 18 pg kg(-1) body weight per day for turkey samples.

This paper reports the results for the surveillance of nine organochlorine pesticides (HCH isomers (alpha, beta, e, gamma), p,p'-DDD, p,p'-DDT, p,p'-DDE, p,p'-DDD, HCB and aldrin) in muscle of three fish species, European pilchard (Sardina pilchardus), Atlantic horse mackerel (Trachurus trachurus) and Atlantic mackerel (Scomber scombrus). Analytical methodology included n-hexane extraction, clean-up with 2% deactivated Florisil, and quantification with gas chromatography-electron capture detection (GC-ECD). The highest mean concentrations were found for p,p'-DDT in sardine and mackerel at levels of 30.1 and 109.9 microg kg(-1), respectively, and for p,p'-DDD in horse mackerel at 51.9 microg kg(-1). Three species had higher levels for S-DDT than S-HCH. The estimated daily intake of organochlorine pesticides in the three species showed that in sardine, the highest EDIs were found for aldrin, at 1.8 ng kg(-1) bw day(-1), which represents 1.8% of the acceptable daily intake (ADI), and for ss-HCH, at 4.0 ng kg(-1) bw day(-1), representing 0.4% of ADI. Lowest values were found for Atlantic mackerel. Statistical analysis to determine the differences in mean concentrations of pesticides between species, and any correlation between groups of residues related with each one of the species, was undertaken.

An analytical method for the determination of OTC and TC residues in honey was developed. Sample treatment involves an extraction in EDTA-McIlvaine buffer, followed by a solid-phase cleanup step. With regard to the cleanup procedure, different SPE cartridges were evaluated and the results presented. The method was validated according to the guidelines laid down by the 2002/657/EC European Decision parameters: decision limit (Ccα) and detection capability (CCβ) were 20 and 21 μg/Kg and 49 and 50 μg/Kg for OTC and TC, respectively, and recoveries of OTC and TC from spiked samples, at three fortification levels, were higher than 87% for both compounds. The analytical method was applied to 57 honey samples. Keywords: Antibiotics; Tetracyclines; Honey; Solid-phase extraction; High-performance liquid chromatography

In this study nine organochlorine pesticide residues ([alpha]-, [beta]-, and [gamma]-hexachlorocyclohexane (HCH), hexachlorobenzene (HCB), aldrin, p,p'-DDE, p,p'-DDD, o,p'-DDT, and p,p'-DDT) in forty nine samples of honey collected from markets of Portugal and Spain during 2001 and 2002, respectively, were evaluated. For this evaluation, three analytical procedures were studied. The analytical procedure, based on LLE extraction with ethyl acetate followed by gas chromatography-electron-capture detection (GC-ECD) for quantification, and mass spectrometry (GC-MS) for confirmation, has been selected. Recoveries of spiked samples ranged from 68%, for [beta]-HCH, and 126% for p,p'-DDT, for fortification levels between 10 and 100 [mu]g/kg, and 64%, for [alpha]-HCH, and 143% for [gamma]-HCH for fortification levels between 20 and 200 [mu]g/kg. Limits of quantification, using GC-ECD, were from 0.01 and 0.10 mg/kg, and limits of detection between 0.001 and 0.02 mg/kg. Fourteen Valencian samples were contaminated, containing residues of HCB or/and HCH isomers. The frequency of detection was 56% for Spanish samples. In Portugal, 23 samples were contaminated, what means 95.8%. In Spanish samples, concentrations range from nd to 0.03 mg/kg for HCB, and nd to 2.24 mg/kg for HCH-total. The mean concentration and standard deviation were 0.017 ± 0.011 mg/kg for HCB, and 0.579 ± 0.747 mg/kg for HCH-total, contributing the [gamma] isomer with the highest values. The samples from Portugal showed higher levels. Levels of HCB ranged from nd to 0.39 mg/kg. HCH-total ranged from nd to 4.86 mg/kg, and DDT-total from nd to 0.658 mg/kg. Mean concentration and standard deviation were 0.09 ± 0.116 mg/kg for HCB, 1.357 ± 1.30 mg/kg for HCH-total, and 0.143 ± 0.193 mg/kg for DDT-total. http://www.sciencedirect.com/science/article/B6TG8-4D75MJM-2/1/228ba9bcfae0d95d011e1a86b66f79dc