Biomonitoring of non-persistent pesticides in urine from lactating mothers: Exposure and risk assessment

Abstract The aim of the present study was to assess the exposure to pesticides in urine from Spanish lactating mothers (n = 116). Six nonspecific (dialkyl phosphates) and 20 specific metabolites of organophosphate pesticides (OPs), herbicides and pyrethroids were analyzed. The most frequently detected biomarkers were diethyl phosphate, p-nitrophenol, 3,5,6-trichloro-2-pyridinol and 3-phenoxybenzoic acid, whose geometric means were 1.9 ng·mL−1, 0.8 ng·mL−1, 1.5 ng·mL−1 and 1.4 ng·mL−1, respectively. Herbicide metabolites were the least frequently detected biomarkers with detection frequencies between 0% (2,4,5-Trichlorophenoxyacetic acid) and 22% (2,4-Dichlorophenoxyacetic acid). Multiple re…

Determination of PAHs in airborne particles by accelerated solvent extraction and large-volume injection-gas chromatography-mass spectrometry.

Abstract A sensitive and automated method is presented for the determination of polycyclic aromatic hydrocarbons (PAHs) in airborne particulate matter. The procedure includes extraction of PM10-bound PAHs by accelerated solvent extraction (ASE) followed by gel permeation chromatography (GPC) clean-up, and large-volume programmable temperature vaporizer (PTV–LV) injection coupled to GC–MS. The limit of detection (LOD) of the whole method, based on a signal-to-noise ratio (S/N) of 3:1, ranged from 0.26 pg m−3 to 3 pg m−3 when air volumes of 760 m3 are collected. The hexane–acetone mixture (1:1, v/v) gave the best recoveries when ASE parameters were fixed at 125 °C, 1500 psi, and a total time …

Application of accelerated solvent extraction followed by gel performance chromatography and high-performance liquid chromatography for the determination of polycyclic aromatic hydrocarbons in mussel tissue.

Accelerated solvent extraction (ASE) has been evaluated as a fast alternative to methanolic saponification for the extraction of 12 polycyclic aromatic hydrocarbons (PAHs) from mussel tissue. Several solvent systems and different operating conditions were investigated. The mixture dichloromethane-acetone (1:1, v/v) gave the best recoveries at 125 degrees C and 1500 psi, in a total time of 10 min. No yield difference was found between freeze-drying (Fd) or drying the wet mussel with diatomaceous earth (Ded) prior to extraction. The ASE method was validated using the standard reference material SRM 2977, a freeze-dried mussel tissue with naturally present organic contaminants. The performance…

Biomonitoring of polycyclic aromatic hydrocarbons in the urine of lactating mothers: Urinary levels, association with lifestyle factors, and risk assessment

Abstract Polycyclic aromatic hydrocarbons (PAHs) are pollutants produced during incomplete combustion of organic matter and several industrial processes. Humans can be exposed to PAHs through ingestion of food, inhalation of tobacco smoke or polluted air, and dermal contact, causing immunologic, developmental, and reproductive problems. In the present research, eleven metabolites of PAHs were analyzed in the urine of 110 lactating women living in Spain (2015). PAH metabolites were extracted from the urine samples by liquid-liquid extraction and their determination was performed by liquid chromatography coupled to tandem mass spectrometry. In addition, information on lifestyle and dietary ha…

Risk assessment and monitoring programme of nitrates through vegetables in the Region of Valencia (Spain)

This study was carried out to determine current levels of nitrate in vegetables marketed in the Region of Valencia (Spain) and to estimate the toxicological risk associated with their intake. A total of 533 samples of seven vegetable species were studied. Nitrate levels were derived from the Valencia Region monitoring programme carried out from 2009 to 2013 and food consumption levels were taken from the first Valencia Food Consumption Survey, conducted in 2010. The exposure was estimated using a probabilistic approach and two scenarios were assumed for left-censored data: the lower-bound scenario, in which unquantified results (below the limit of quantification) were set to zero and the up…

Exposure and cumulative risk assessment to non-persistent pesticides in Spanish children using biomonitoring.

Abstract The main objective of the present research is to evaluate the exposure to pesticides in children (n = 568) from the Valencian Region (Spain). Six non-specific and 20 specific metabolites of organophosphate pesticides (OPs), herbicides (Herb), and pyrethroids (Pyr) were analyzed in urine samples. The biomarkers with the highest detection frequencies (>70%) were diethyl phosphate, p-nitrophenol, 3-phenoxybenzoic acid, and 3,5,6-trichloro-2-pyridinol, whose geometric mean concentrations (ng·ml-n1) were 1.53, 1.03, 1.51 and 1.19, respectively. Robust regression models showed that the province of residence and the recent consumption of vegetables, legumes and cereals were some of the mo…

Determination of 3-MCPD by GC-MS/MS with PTV-LV injector used for a survey of Spanish foodstuffs

3-Monochloropropane-1,2-diol (3-MCPD) is the most common chemical contaminant of the group of chloropropanols. It can occur in foods and food ingredients at low levels as a result of processing, migration from packaging materials during storage and domestic cooking. A sensitive method for determination of 3-MCPD in foodstuffs using programmable temperature vaporization (PTV) with large-volume injection (LVI) gas chromatography (GC) with tandem mass spectrometry detection (MS/MS) has been developed and optimized. The optimization of the injection and detection parameters was carried out using statistical experimental design. A Plackett-Burman design was used to estimate the influence of reso…

Biomonitoring of bisphenols A, F, S in human milk and probabilistic risk assessment for breastfed infants

The present study addresses the presence of bisphenols A (BPA) and its analogs bisphenol F (BPF) and S (BPS) in milk of 120 mothers living in Valencia (Spain) and participating in the BETTERMILK project (year 2015). We also studied the factors that could influence the BPA levels and estimated the exposure and the risk for breast fed infants. The frequency of detection of total (conjugated + unconjugated) and unconjugated-BPA were 83% and 77%. with a geometric mean of 0.29 ng/mL and 0.15 ng/mL, respectively. The frequency of detection was much lower for total-BPF (22%) and total-BPS (1.1%). The place of residence of the mother and the use of personal care products showed significant associat…

Risk assessment of exposure to phthalates in breastfeeding women using human biomonitoring

In this study, we assessed the presence of 14 phthalate metabolites in the urine of 104 lactating mothers from Valencia (Spain) who took part in the human biomonitoring project BETTERMILK. Nine of the metabolites studied showed detection frequencies >80%, whereas the rest of the metabolites presented low detection frequencies (<5%). The concentrations ranged from <LoQ to 1291 ng/mL with monoethyl phthalate showing the highest concentration, with a geometric mean of 34.90 ng/mL. In general, the phthalate metabolite levels quantified in the present study were lower than the urinary levels found in previous studies that involved lactating mothers. The consumption of packaged juices and the fre…

Dietary exposure to trace elements and health risk assessment in the Region of Valencia (Spain). A Total Diet Study

ABSTRACTDietary exposure of the Valencian region population to lead, cadmium, inorganic arsenic (iAs), chromium, copper, tin and methylmercury (meHg) was assessed in a total diet study carried out in the region of Valencia in 2010–11. A total of 8100 food samples were collected and analysed. Occurrence data were combined with consumption data to estimate dietary exposure in adults (> 15 years of age) and young children (6–15 years of age). The estimated intake was calculated by a probabilistic approach. Average intake levels (optimistic scenario) for lead, iAs, chromium and tin were 0.21, 0.08, 1.79 and 1.87 µg kg−1 bw day−1 respectively; for Cd and meHg average intake levels were 0.77 and …

Determination of acrylamide in foods by pressurized fluid extraction and liquid chromatography-tandem mass spectrometry used for a survey of Spanish cereal-based foods.

An automated and rapid method for the determination of acrylamide in different food products is presented. The method involves pressurized fluid extraction (PFE) of foods with acetonitrile and precipitation with Carrez reagents. The final extract is analysed by liquid chromatography coupled to electrospray ionization tandem mass spectrometry (ESI-MS-MS). The main parameters affecting the performance of ESI-MS-MS and PFE were optimized using a design of experiments approach. The limit of quantification of the method was 5 microg kg(-1), and recoveries from incurred samples ranged between 93 and 101%. The accuracy was evaluated using the reference test materials FAPAS T3002, T3005 and T3011. …

Occurrence of biomarkers of pesticide exposure in non-invasive human specimens.

Biomonitoring has been used in many types of investigations, including national programmes and epidemiological studies, to explore the occurrence of biomarkers of pesticide exposure in the general population or relevant groups. This review discusses recent studies that measure levels of biomarkers of pesticide exposure in non-invasive human specimens such as urine, breast milk, meconium and hair. Specific and non-specific metabolites of organophosphate and pyrethroid insecticides have been widely investigated in urine, where some of the suitable biomarkers present rates of detection higher than 80%, which stand for an ongoing chronic exposure to traces of these chemicals. Hair is a promisin…

Biomonitoring of parabens in human milk and estimated daily intake for breastfed infants

In this study, we assessed the presence of four parabens in human milk of 120 mothers from Valencia (Spain) which took part in a human biomonitoring project (BETTERMILK). The detection frequency ranges of parabens were 41-60% and 61-89% for unconjugated- and total (unconjugated + conjugated)-parabens, respectively. The concentrations ranged from <LoQ to 31 ng/mL and from <LoQ to 49 ng/mL for unconjugated- and total-parabens, respectively. The frequency of use of some cosmetic products and human milk protein levels were the main predictors of parabens in milk. The study evidences the presence of both conjugated and unconjugated paraben forms in human milk. The newborns estimated daily intake…

Biomonitoring of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and dioxin-like polychlorinated biphenyls (dl-PCBs) in human milk: Exposure and risk assessment for lactating mothers and breastfed children from Spain

Abstract The main objective of the present research was to evaluate the levels of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and dioxin-like polychlorinated biphenyls (dl-PCBs) in the human milk of Spanish lactating women who participated in the BETTERMILK project so that exposure and risk could be assessed for these mothers and their breastfed children. The total ∑PCDD/Fs + dl-PCBs TEQ2005 concentrations in the upper-bound (UB) ranged from 1.29 to 13.48 pg TEQ2005 g−1 lipid. The estimated geometric mean level for the sum of PCDD/F and dl-PCBs was 4.10 and 4.42 pg TEQ2005 g−1 lipid, lower-bound (LB) and UB respectively and were below the reference leve…

Determination of bisphenol diglycidyl ether residues in canned foods by pressurized liquid extraction and liquid chromatography–tandem mass spectrometry

Abstract A new confirmatory method for simultaneous determination of bisphenol diglycidyl ether residues (BADGE, BADGE·H2O, BADGE·2H2O, BADGE·H2O·HCl, BADGE·HCl, BADGE·2HCl, BFDGE and BFDGE·2HCl) from canned food has been developed. The proposed method includes extraction by pressurized liquid extraction (PLE) followed by liquid–liquid partition and purification by solid phase extraction (SPE). Several solvent systems and different operating conditions (time, temperature) have been investigated for PLE optimization. A reversed-phase high-performance liquid chromatography (RP-HPLC) coupled to atmospheric pressure chemical ionisation tandem mass spectrometry (APCI-MS–MS) method was developed …

Optimization of a microwave-assisted extraction large-volume injection and gas chromatography–ion trap mass spectrometry procedure for the determination of polybrominated diphenyl ethers, polybrominated biphenyls and polychlorinated naphthalenes in sediments

Abstract A sensitive and rapid method for the determination of polybrominated diphenyl ethers (PBDEs), polybrominated biphenyls (PBBs) and polychlorinated naphthalenes (PCNs) in sediment samples is proposed. The method involving microwave-assisted extraction (MAE) and large-volume injection (LVI) gas chromatography (GC)–ion trap mass spectrometry (ITMS), and the experimental conditions were optimized using the statistical design of experiments (DOE). A Plackett–Burman (P–B) design was chosen to estimate the influence of five factors, such as resonance excitation voltage (EV), isolation time (IT), excitation time (ET), ion source temperature (IST) and electron energy (EE) on the analytical r…

Biomonitoring of Phthalates, Bisphenols and Parabens in Children: Exposure, Predictors and Risk Assessment

Exposure to emerging contaminants, such as phthalates, bisphenols and parabens in children has been associated with possible neurodevelopment and endocrine alterations. In the present study, the biomonitoring of biomarkers in children (5–12 years old) from the Valencia Region (Spain) have been implemented using urines from the BIOVAL program. More than 75% of the children studied (n = 562) were internally exposed (&gt

Development of a pressurised liquid extraction and liquid chromatography with electrospray ionization-tandem mass spectrometry method for the determination of domoic acid in shellfish.

Amnesic shellfish poisoning (ASP) is a potentially lethal human toxic syndrome which is caused by domoic acid (DA) that originates in marine phytoplankton belonging to the Pseudonitzschia genus. A confirmatory and sensitive procedure has been developed and validated for the determination of DA in shellfish. The proposed method includes pressurised liquid extraction (PLE) with methanol/acetone (9:1), florisil ® cleanup purification inside the PLE extraction cell and detection by liquid chromatography (LC) coupled to electrospray ionization in positive mode tandem mass spectrometry (ESI-MS–MS). Comparison of ionization sources (ESI, atmospheric pressure ionization (APCI) atmospheric pressure …

Assessment of metal levels in foodstuffs from the Region of Valencia (Spain).

Graphical abstract

Biomonitoring of 20 elements in urine of children. Levels and predictors of exposure

The levels of 20 elements in the urine of one hundred twenty children (Ages 6-11) from an agricultural and an urban area of Valencia Region, Spain, were assessed. The seven essential elements analysed (Co, Cu, Mn, Mo, Se, V, Zn) presented average concentrations (geometric mean) ranging from 0.22 µg L(-1) to 63 µg L(-1) (except Zn with 525 µg L(-1)). Eight out of the 13 toxic elements determined (As, Ba, Cd, Cs, Ni, Pb, Tl, Hg) had frequencies of detection of 100%, with geometric mean concentrations from 0.17 µg L(-1) (Cd) to 33.6 µg L(-1) (As). About 18% of children revealed urinary levels of total As higher than the normal range of 100 µg L(-1) accepted by the ATSDR. The urine analysis was…

Protein Biomarkers of Bovine Defective Meats at a Glance: Gel-Free Hybrid Quadrupole-Orbitrap Analysis for Rapid Screening

An understanding of biological mechanisms that could be involved in the stress response of animal cattle prior to slaughter is critical to create effective strategies aiming at the production of high-quality meat. The sarcoplasmic proteome of directly extracted samples from normal and high ultimate pH (pHu) meat groups was studied through a straightforward gel-free strategy supported by liquid chromatography hybrid quadrupole-Orbitrap high-resolution mass spectrometry (LC-HRMS) analysis. A stepped proteomic pipeline combining rapid biomarker hunting supported by qualitative protein Mascot scores followed by targeted label-free peptide quantification revealed 26 descriptors that characterize…

Probabilistic risk assessment of the exposure to polybrominated diphenyl ethers via fish and seafood consumption in the Region of Valencia (Spain)

The study was carried out to estimate the dietary intake of polybrominated diphenyl ethers (PBDEs) in the Region of Valencia (Spain) in order to evaluate the resultant risk. The PBDE levels in fish and seafood (a total of 206 samples) were determined. Dietary intake was estimated using results of PBDE analyses in fish and seafood marketed in the Region of Valencia over the period 2007-2012 and data of the first Valencian Food Consumption Survey, conducted in 2010. Two scenarios were assumed for left-censored results: the lower-bound (LB) scenario, in which unquantified results (below the limit of quantification (LOQ)) were set to zero and the upper-bound (UB) scenario, in which unquantified…

Determination of 21 perfluoroalkyl substances and organophosphorus compounds in breast milk by liquid chromatography coupled to orbitrap high-resolution mass spectrometry.

Abstract The human exposure to perfluoroalkyl (PFASs) and organophosphorus (OPs) compounds is a cause of concern and its determination in biological matrices, including human milk, is an emergent approach for evaluate their exposure. A comprehensive strategy for the quantitative determination of 21 PFASs and OPs compounds in breast milk was developed. The proposed method includes an extraction and clean-up procedure based on the QuEChERs methodology followed by an ultra-high performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS) determination. The full-scan mass data were acquired with a resolution of 50000 FWHM and a mass accuracy better than 5 ppm. Met…

Dietary exposure and risk assessment of polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans and dioxin-like polychlorinated biphenyls of the population in the Region of Valencia (Spain)

Dietary exposure of the Valencia Region population to polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and PCBs was assessed in the Region of Valencia in 2010–2011. A total of 7700 food samples were collected. Occurrence data were combined with consumption data to estimate dietary exposure in adults (&gt;15 years of age) and young people (6–15 years of age). The estimated intake was calculated by a probabilistic approach. Average intake levels (upper-bound scenario) were 1.58 and 2.76 pg toxic equivalent (TEQ) kg−1 body weight (bw) day−1 for adults and young people, respectively. These average intakes are within range of the tolerable daily intake of 1–4 pg W…

Children’s exposure to polycyclic aromatic hydrocarbons in the Valencian Region (Spain): Urinary levels, predictors of exposure and risk assessment

Polycyclic aromatic hydrocarbons (PAHs) are pollutants that are released into the environment during incomplete combustion of organic matter and which can have a negative effect on human health. PAHs enter the human body mostly through ingestion of food or inhalation of tobacco smoke. The purpose of the present study is to evaluate the internal levels of PAHs that children living in the Valencian Region (Spain) are exposed to. In total, we measured eleven biomarkers of exposure to naphthalene, fluorene, phenanthrene, pyrene, and benzo(a)pyrene in the urine of 566 children aged 5-12. The analytical method was based on a liquid-liquid extraction of the PAH metabolites from the urine samples, …

Polybrominated diphenyl ethers in foods from the Region of Valencia: Dietary exposure and risk assessment

Abstract Dietary exposure to polybrominated diphenyl ethers (PBDEs) of the population in the Region of Valencia, Spain, was assessed. A group of 320 composite samples of different fatty foods was collected and analyzed, including the following: vegetable oils, and foods of animal origin such as (a) fish and seafood, (b) eggs, (c) milk and dairy products, and (d) meat and meat products. Two scenarios were assumed for left-censored results: lower-bound (LB) and upper-bound (UB). Vegetable oils, and fish and seafood presented the highest content of PBDEs [mean values of 503 and 464 pg g−1 wet weight (ww) for total PBDEs, respectively, in the UB]. The dominating congeners were BDE47 in the food…

Human risk assessment and regulatory framework for minerals in food

Exposure assessment of Spanish lactating mothers to acrylamide via human biomonitoring.

Abstract Acrylamide (AA) is an organic compound classified as “Probably carcinogenic to humans” (Group 2 A) that can be found principally in processed carbohydrate-rich foods and tobacco smoke. In humans, after exposure, AA is rapidly metabolized and excreted in urine, predominantly as N-acetyl-S-(2-carbamoylethyl)-l-cysteine (AAMA), N-acetyl-S-(2-carbamoyl-2-hydroxyethyl)-l-cysteine (GAMA3) and N-Acetyl-3-[(3-amino-3-oxopropyl)sulfinyl]-L-alanine (AAMA-Sul), which can be used as short-term biomarkers of exposure to AA. In this study, the presence of AA metabolites in urine samples of lactating mothers living in Spain (n = 114) was analyzed by “dilute and shoot” and liquid chromatography co…

Exposure and risk assessment to arsenic species in Spanish children using biomonitoring

We present a new approach to arsenic (As) risk assessment using biomonitoring. In this pilot study we determined the levels of total and speciated urinary arsenicin 109 Spanish school children aged between 6 and 11 years, and interpreted these concentrations in a risk assessment context. The geometric mean (GM) for total As (TAs) was 33.82 mu/L. The order of occurrence and average concentrations of the different species was arsenobetaine (AsB) (100%, 15 mu g/L), dimethylarsinic (DMA) (97%, 8.32 mu g/L), monomethylarsonic (MMA) (26%, 027 mu g/L) and inorganic As (iAs) (4%, 0.14 mu g/L). 18% of children presented exposures to inorganic arsenic (7.52 mu g/g creatinine) higher than guidance val…

Determination of acrylamide in coffee and chocolate by pressurised fluid extraction and liquid chromatography-tandem mass spectrometry.

A selective and sensitive procedure has been developed and validated for the determination of acrylamide in difficult matrices, such as coffee and chocolate. The proposed method includes pressurised fluid extraction (PFE) with acetonitrile, florisil clean-up purification inside the PFE extraction cell and detection by liquid chromatography (LC) coupled to atmospheric pressure ionisation in positive mode tandem mass spectrometry (APCI-MS-MS). Comparison of ionisation sources (atmospheric pressure chemical ionisation (APCI), atmospheric pressure photoionization (APPI) and the combined APCI/APPI) and clean-up procedures were carried out to improve the analytical signal. The main parameters aff…

Development of a novel methodology for determination of dialkyl phosphates in human urine using liquid chromatography-tandem mass spectrometry

Abstract In this study a fast and sensitive method was developed for the quantitative determination of six dialkyl phosphates (DAPs) in human urine using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The proposed methodology was based on liquid-liquid extraction (LLE) with diethyl ether and ethyl acetate using MgSO4 and NaCl, followed by liquid chromatography and mass spectrometry detection in the selected reaction mode (SRM) with negative electrospray ionization (ESI-). Detection settings were optimized by design of experiments (DoE). Dibutyl phosphate (DBP) was used as internal standard. Several criteria established in the SANTE/11813/2017 guidance document for pesticide resi…

Determination of four parabens and bisphenols A, F and S in human breast milk using QuEChERS and liquid chromatography coupled to mass spectrometry

Abstract For the first time, a multiresidue, sensitive and high throughput method for determination of bisphenol A, F and S and 4 parabens (methyl paraben, ethyl paraben, propyl paraben and butyl paraben) in human breast milk was developed. The proposed method includes an extraction and clean-up procedure based on QuEChERS methodology followed by liquid chromatography coupled to triple quadrupole mass spectrometry determination. Negative atmospheric pressure chemical ionization in the selected reaction monitoring mode was used for mass detection. During the method validation the recoveries varied between 83 and 115% with a precision lower than 20% for all analytes using spiked levels from 0…

Chronic cumulative risk assessment of the exposure to organophosphorus, carbamate and pyrethroid and pyrethrin pesticides through fruit and vegetables consumption in the region of Valencia (Spain).

In the present study, the chronic cumulative exposure to organophosphorus (OPs), carbamates (CBs) and pyrethroid and pyrethrin (PPs) pesticides in the region of Valencia through fruit and vegetables consumption is presented. A total of 752 samples and 84 pesticides were studied of which, 52 were OPs, 23 CBs and 9 PPs. Residue data were derived from the Valencia Region monitoring program 2007-2011 and food consumption levels from a questionnaire-based dietary survey conducted in 2010 in the same area. The relative potency factor (RPFs) approach was used to estimate chronic cumulative dietary exposure to OPs, CBs and PPs using acephate, oxamyl and deltamethrin as index compounds, respectively…

Development of a method for the analysis of seven banned azo-dyes in chilli and hot chilli food samples by pressurised liquid extraction and liquid chromatography with electrospray ionization-tandem mass spectrometry

An automated, confirmatory and sensitive procedure has been developed and validated for the determination of Sudan (I-IV), Sudan Orange G, Sudan Red 7B and Para Red in hot chilli food samples. The proposed method includes pressurised liquid extraction (PLE) with acetone, gel permeation chromatography (GPC) clean-up and detection by liquid chromatography (LC) coupled to electrospray ionization in positive mode tandem mass spectrometry (ESI-MS-MS). The main parameters affecting the performance of the different ionization sources and PLE parameters were previously optimised using statistical design of experiments (DOE). The method was in-house validated on chilli powder and chilli meat. Linear…

Dietary exposure to trace elements and health risk assessment in the region of Valencia, Spain: a total diet study

Dietary exposure of the Valencian region population to lead, cadmium, inorganic arsenic (iAs), chromium, copper, tin and methylmercury (meHg) was assessed in a total diet study carried out in the region of Valencia in 2010–11. A total of 8100 food samples were collected and analysed. Occurrence data were combined with consumption data to estimate dietary exposure in adults (&gt; 15 years of age) and young children (6–15 years of age). The estimated intake was calculated by a probabilistic approach. Average intake levels (optimistic scenario) for lead, iAs, chromium and tin were 0.21, 0.08, 1.79 and 1.87 µg kg−1 bw day−1 respectively; for Cd and meHg average intake levels were 0.77 and 0.54 …