Biomonitoring of common organophosphate metabolites in hair and urine of children from an agricultural community

Levels of dialkylphosphate (DAP) metabolites were measured in hair and urine of children that lived close to intensively farmed areas of Almeria (Southeast Spain). The levels were used as proxies for exposure of these children to organophosphate pesticides (OPs). Determinants of exposure to DAPs were also examined. Urine and hair samples were collected from 222 children aged 3–11 years and information on lifestyle and dietary habits was collected from questionnaires administered to mothers. Urinary DAPs were analyzed by ultra-high performance liquid-chromatography coupled to triple-quadrupole tandem mass-spectrometry (UHPLC-QqQ-MS/MS) and hair DAPs by gas-chromatography coupled to mass spectrometry (GC–MS). Detection rates ranged from 21.8% for diethylphosphate (DEP) and diethylthiophosphate (DETP) to 35.9% for dimethylphosphate (DMP) in urine; and from 42.3% for DETP to 92.8% for DMP in hair. Diethyldithiophosphate (DEDTP) was detected in 0.5% of urine samples (one child), and in 26.6% of children's hair samples. A lack of correlation was observed for individual DAP metabolites and ΣDAPs between urine and hair samples, except for DEDTP. Urinary DAP levels of our child population were lower than those reported for children from other countries, including NHANES 1999–2000 data.This study was partially funded by the Ministry of Economy, Innovation, Science and Employment, Government of Andalucia (reference number P08-CTS-04313, FEDER funds)

Development of mass spectrometry analytical methods for the determination of pyrethroid and neonicotinoid pesticides in laboratory animal biological samples. Implementation of them for the biomonitoring of rural and special population groups

Aim of this study was to develop and validate new analytical methods for thedetection and quantification of parent and metabolite compounds of cypermethrinand imidacloprid pesticides in various biological samples. Also their toxicity wasinvestigated in-vivo. Pyrethroids and neonicotinoids are two classes of pesticideswidely and extensively used the last decades. The interest concerning thesesubstances is growing, since they are used as replacement for the organophosphorusand carbamates pesticides. Of a great concern is also the biomonitoring ofpopulation to these chemicals, especially for occupationally exposed individuals whoare exposed longer and in higher levels. Cretan population consists of a mixture ofrural and urban residents, while many individuals dealing with agricultural activitiesas amateurs. Furthermore, the residence of cretan population is offen nearagricultural facilities such as open fields and green houses. This results multiple riskand burden to the aforementioned pesticides. In literature even increasing reports ofacute or chronic toxicity cases of these substances can be found. The escalatingconcerns of the international scientific community on the effects of these substanceson populations, gave rise to this study.Cypermethrin is a synthetic pyrethroid pesticide used for both domestic andagricultural applications. Its chemical structure is relative to the natural pyrethrin. Itis supposed to be relatively low toxic for human and other mammals, while acts asneurotoxin when contact with insects. The specific metabolites for cypermethrin arecis- and irons-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carboxylic acids (cisandtrans-Cl2CA) and the non-specific metabolite 3-phenoxybenzoic acid (3-PBA),which is common for at least ten different pyrethroids. Neonicotinoids are also agroup of synthetic pesticides with insecticidal act that found a lot of applicationmainly in agriculture, the last three decades. Imidacloprid is one of the mostcommon neonicotinoid, which is metabolized to the non-specific 6-chloronicotinicacid (6-ClNA). Although limited references can be found concerning neonicotinoids,it is proven that the toxicity of these pesticides must be examined. In-vivo experiments were executed on rabbits. Laboratory animals of samegender and age were used and were exposed orally with sub-acute dosages of theaforementioned pesticides. The administrated levels were calculated in order torepresent the 1/15 and 1/30 of the LD50 (lethal dose) when it was possible. Durationof the experiment was about 6 months. During experiment, samplings of biologicalsamples (urine, hair, blood) were executed (before beginning the administration andafter completing two months).Analytical methods for the detection of the parent compound and themetabolites were developed, using techniques of chromatography and massspectrometry. The developed methods were validated by calculating severalanalytical parameters (linearity, recovery, precision, accuracy, limits of detection-LOD, limits of quantification-LOQ). Furthermore, the results were compared withcontamination parameters (administrative dosage, duration of exposure).Briefly, for the detection of imidacloprid and its metabolite 6-ClNA, ananalytical method using liquid chromatography-mass spectrometry was developed.Limit of detection for imidacloprid was 0.02 ng/mg and 0.01 ng/mg for 6-ClNA in hairsamples, while in urine samples the corresponding values were 0.002 μg/ml forimidacloprid and 0.008 μg/ml for 6-ClNA. Satisfactory chromatographic separationwas achieved with retention time for imidacloprid 11.13 min and 12.46 min for 6-ClNA. Linearity of both standard solutions and spiked samples was more thansatisfactory for both hair and urine developed methods (>0.99). Mean recovery forimidacloprid was calculated to be 97.13% (n=9) and 91.42% (n=9) for 6-ClNA in hairsamples, while in urine samples the corresponding values were 96.11% (n=9) and103.44% (n=9). The method accuracy was >98.71% for imidacloprid and >100.25%for 6-ClNA in hair samples method and >98.93% for imidacloprid and >97.82% for 6-ClNA in urine samples method. The corresponding precision values were fluctuated0.99). Meanrecovery for cis-Cl2CA was 84.8% (n=8), for trans-Cl2CA 87.2% (n=8) and for 3-PBAwas 96.4% (n=8) for hair samples, while for urine samples the correspondingrecoveries were 102.1% (n=3), 111.2% (n=3) and 87.1% (n=3). Accuracy wasestimated to be >91.8% for cis-Cl2CA, >96.7% for trans-Cl2CA and >90.5% for 3-PBAfor hair samples method and >89.2% for cis-Cl2CA, >71.6% for trans-Cl2CA and>86.0% for 3-PBA for urine samples method. The corresponding precision valueswere fluctuated 0.99).Η μέση ανάκτηση για το imidacloprid ήταν 97.13% (n=9) και 91.42% (n=9) για το 6-ClNA για τα δείγματα τριχών, ενώ για τα δείγματα των ούρων οι αντίστοιχες ανακτήσεις ήταν 96.11% (n=9) και 103.44% (n=9). Η ακρίβεια της μεθόδου(accuracy) ήταν >98.71% για το imidacloprid και >100.25% για το 6-ClNA για τοπρωτόκολλο της τρίχας και >98.93% για το imidacloprid και >97.82% για το 6-ClNAγια το πρωτόκολλο των ούρων. Οι αντίστοιχες τιμές της ακρίβειας (precision)κυμαίνονταν 0.99). Η μέση ανάκτηση για το cis-Cl2CA ήταν 84.8% (n=8), για το trans-Cl2CA 87.2% (n=8) και για το 3-PBA ήταν 96.4%(n=8) για τα δείγματα τριχών, ενώ για τα δείγματα των ούρων οι αντίστοιχεςανακτήσεις ήταν 102.1% (n=3), 111.2% (n=3) και 87.1% (n=3). Η ακρίβεια τηςμεθόδου (accuracy) υπολογίστηκε >91.8% για το cis-Cl2CA, >96.7% για το trans-Cl2CAκαι >90.5% για το 3-PBA για το πρωτόκολλο της τρίχας και >89.2% για το cis-Cl2CA,>71.6% για το trans-Cl2CA και >86.0% για το 3-PBA για το πρωτόκολλο των ούρων.Οι αντίστοιχες τιμές της ακρίβειας (precision) κυμαίνονταν <13.6% για όλες τιςπεριπτώσεις.Τα δείγματα τριχών των πειραματόζωων που αναλύθηκαν έδωσανστατιστικά σημαντικές διαφορές ανάμεσα στις ομάδες των ζώων (control, χαμηλήδόση, υψηλή δόση) για το διάστημα 0 έως 4 μήνες έκθεσης για όλους τουςμεταβολίτες, cis-Cl2CA (p=0.009), trans-Cl2CA (p=0.003), 3-PBA (p=0.012). Οισυγκεντρώσεις του cis-Cl2 CA στα δείγματα τριχών για την ομάδα ελέγχου (control)κυμαίνονταν από <LOD έως 112.6 pg/mg, για την ομάδα της χαμηλής δόσης από<LOD έως 1022.0 pg/mg και για την ομάδα της υψηλής δόσης από <LOD έως 5361.4pg/mg. Οι αντίστοιχες συγκεντρώσεις του trans-Cl2CA στα δείγματα τριχών για τηνομάδα ελέγχου (control) κυμαίνονταν από <LOD έως 112.0 pg/mg, για την ομάδατης χαμηλής δόσης από <LOD έως 1367.0 pg/mg και για την ομάδα της υψηλήςδόσης από <LOD έως 5448.2 pg/mg. Τέλος για το 3-PBA οι συγκεντρώσεις σταδείγματα τριχών για την ομάδα ελέγχου (control) κυμαίνονταν από 16.5 έως 42.8pg/mg, για την ομάδα της χαμηλής δόσης από <LOD έως 146.7 pg/mg και για την ομάδα της υψηλής δόσης από <LOD έως 659.9 pg/mg. Τα αποτελέσματα πουπροέκυψαν από την ανάλυση των τριχών των πειραματόζωων έδειξαν αύξηση τωνεπιπέδων όλων των μεταβολιτών με τη πάροδο του χρόνου και η αύξηση αυτή ήτανανάλογη της χορηγούμενης δόσης.Οι συγκεντρώσεις όλων των μεταβολιτών στα δείγματα ούρων για τηνομάδα ελέγχου (control) ήταν <LOD για όλη τη διάρκεια του πειράματος. Για τηνομάδα της χαμηλής δόσης οι συγκεντρώσεις για το cis-Cl2CA από <LOD έως 1365.6ng/ml και για την ο

Cardiotoxicity in rabbits after long-term nandrolone decanoate administration

Abuse of anabolic androgenic steroids is linked to a variety of cardiovascular complications. The aim of our study was to investigate the possible cardiovascular effects of nandrolone decanoate on young rabbits using echocardiography, histology and monitoring of telomerase activity, oxidative stress and biochemical markers. Fourteen rabbits were divided into three administration groups and the control group. Doses of 4 mg/kg and 10 mg/kg of nandrolone decanoate, given intramuscularly and subcutaneously, two days per week for six months were applied. A 4-months wash-out period followed. Focal fibrosis and inflammatory infiltrations of cardiac tissue were observed in the high dose groups. Thiobarbituric acid-reactive species (TBARS) levels were significantly increased in the high dose groups, while catalase activity decreased. Myocardial Performance Index (MPI) is the main echocardiographic index primarily affected by nandrolone administration in rabbits. Despite the preserved systolic performance, histological lesions observed associated with distorted MPI values, point to diastolic impairment of the thickened myocardium due to nandrolone treatment. Oxidative stress accumulates and telomerase activity in cardiac tissue rises. Subcutaneous administration seems to be more deleterious to the cardiovascular system, as oxidative stress, telomerase activity and biochemical markers do not appear to return into normal values in the wash-out period. (C) 2015 Elsevier Ireland Ltd. All rights reserved

Достижения в области биомониторинга фосфорорганических и хлорорганических пестицидов

Organophosphorus (OPs) and organochlorine (OCs) pesticides are synthetic organic compounds used mainly in agriculture to manage pest problems. Human is occupationally and environmentally exposed to pesticides via inhalation, ingestion and skin absorption. When the pollutant enters human body it is rapidly metabolized to more toxic compounds than the parent compound. Their ability to interfere with biochemical and metabolic pathways in humans makes them compounds of high concern. Acute and chronic exposure has been linked with neurological, cardiovascular, birth effects and male infertility. Human exposure to pesticides is estimated by measuring their metabolites in biological tissues and fluids such as blood, urine, amniotic fluid and hair. In this review study advances on biomonitoring of pesticides are presented. Measured levels of pesticides in urine matrices, amniotic fluid, meconium and hair are presented in order to monitor the human burden and biomonitoring data between occupationally and environmentally exposed populations are being comparedФосфорорганические и хлорорганические пестициды – это синтетические органические соединения, используемые преимущественно в сельском хозяйстве для борьбы с вредителями. Люди подвергаются воздействию пестицидов в процессе профессиональной деятельности и при контакте с зараженной средой через их вдыхание, потребление с пищей и поглощение кожей. При попадании в организм человека поллютант быстро метаболизируется до более токсичных соединений, чем исходное вещество. Их способность взаимодействовать с биохимическими и метаболическими путями в человеческом организме делает их потенциально опасными соединениями. Обнаружена связь между острым и хроническим воздействием пестицидов и неврологическими и кардиоваскулярными эффектами, врожденными отклонениями и мужским бесплодием. Оценка воздействия пестицидов на организм человека проводится путем измерения их метаболитов в биологических тканях и жидкостях, таких как кровь, моча, амниотическая жидкость, а также в волосах. В данном обзоре представлены достижения в области биомониторинга пестицидов. Для оценки нагрузки на человека приведены измеренные уровни пестицидов в моче, амниотической жидкости, меконии и волосах. Проведено сравнение популяций, подвергшихся воздействию пестицидов в связи с производственной деятельностью и при контакте с окружающей средо

Chemical analysis and hazard identification of the most common electronic cigarette liquids in nine European countries

Background We aimed to detect the composition and reported chemical health hazards of the most common electronic cigarette liquids (e-liquids) in nine European Union (EU) Member States (MS) prior to adoption of the Tobacco Product Directive (TPD). Material and Methods Within the Horizon2020, EUREST-PLUS study, 122 of the most commonly used e-liquids were purchased from 9 EU MS. Chromatography - mass spectrometry and liquid chromatography - mass spectrometry methods were used to analyze the samples. Among the most frequently detected compounds (detected ≥4 times), Danger Globally Harmonized System of Classification and Labelling of Chemicals (GHS) and Warning GHS codes were identified. Results During the samples analysis, several discrepancies in nicotine concentration were detected among the samples from the 9 EU MS. French samples contained an average of 21.9% more nicotine than labelled, while Romanian samples contained an average of 22.5% less nicotine than labelled. In addition, in the 9.8% of the samples, the nicotine concentration exceeded the limit of 20 mg/ml. With regards to the samples’ composition, 171 different compounds were identified and detected 750 times in total while we did not identify samples positive for PAHs or nitrosamines. Finally from the 171 substances, only 5 (10.4%), (Oxime-, methoxy-phenyl, +/-.-.alpha.-Methylbenzyl acetate, 1,3-Dioxolane, 2-butyl-4-methyl-, Melonal and l-Menthyl acetate) were not associated with a Danger GHS and Warning GHS codes. Conclusions As large number of potential harmful compounds was identified, the systematic monitoring and chemical evaluation of e-liquids is necessary in order to protect the consumers’ health. Acknowledgements EUREST-PLUS is a Horizon2020 project conducted by researchers throughout Europe from both the six participating countries as well as other institution partners within Europe and abroad. Partnering organizations include the European Network on Smoking Prevention (Belgium), Kings College London (United Kingdom), German Cancer Research Centre (Germany), University of Maastricht (The Netherlands), University of Athens (Greece), Aer Pur Romania (Romania), European Respiratory Society (Switzerland), the University of Waterloo (Canada), the Catalan Institute of Oncology (Catalonia, Spain), Smoking or Health Hungarian Foundation (Hungary), Health Promotion Foundation (Poland), University of Crete (Greece), and Kantar Public Brussels (Belgium). Funding The EUREST-PLUS Project takes place with the financial support of the European Commission, Horizon 2020 HCO-6-2015 program (EUREST-PLUS: 681109; C. Vardavas) and the University of Waterloo (GT. Fong). Additional support was provided to the University of Waterloo by the Canadian Institutes of Health Research (FDN-148477). GT. Fong was supported by a Senior Investigator Grant from the Ontario Institute for Cancer Research. E. Fernández is partly supported by Ministry of Universities and Research, Government of Catalonia (2017SGR139) and by the Instituto Carlos III and co-funded by the European Regional Development Fund (FEDER) (INT16/00211 and INT17/00103), Government of Spain

Antibiotics in Raw Meat Samples: Estimation of Dietary Exposure and Risk Assessment

The extensive use of antibiotics in livestock farming poses increased concerns for human health as residues of these substances are present in edible tissues. The aim of this study was the determination of the levels of four groups of antibiotics (sulfonamides&mdash;SAs, tetracyclines&mdash;TCs, streptomycines&mdash;STr and quinolones&mdash;QNLs) in meat samples (muscles, livers and kidneys from beef, chicken and pork) and the estimation of the dietary exposure to antibiotics from meat consumption and the potential hazard for human health. Fifty-four samples of raw meat were randomly collected in 2018 from the Cretan market, Greece and analyzed both with an enzyme-linked immunosorbent assay (ELISA) and liquid chromatography&ndash;mass spectrometry (LC&ndash;MS). According to the results derived from the ELISA method, only 2% of the meat samples were free from antibiotics, 2% were detected with 4 antibiotics and the great majority of the samples (87%) were detected with 2 to 3 antibiotics. SAs presented the highest detection frequencies for all samples whereas TCs were not detected in any bovine sample. The highest median concentration was detected for STr in bovine muscles (182.10 &mu;g/kg) followed by QNLs (93.36 &mu;g/kg) in pork kidneys whereas the chicken samples had higher burdens of QNLs compared to the other meat samples. LC&ndash;MS analysis showed that oxytetracycline (OTC) was the most common antibiotic in all samples. The highest median concentration of all antibiotics was detected for doxycycline (DOX) (181.73 &mu;g/kg in pork kidney) followed by OTC in bovine liver (74.46 &mu;g/kg). Risk characterization was applied for each of the two methods; The hazard quotients (HQ) did not exceed 0.059 for the ELISA method and 0.113 for the LC&ndash;MS method for any group of antibiotics, whereas the total hazard indexes (HI) were 0.078 and 0.021, respectively. The results showed the presence of different groups of antibiotics in meat from the Cretan market and that the health risk to antibiotics is low. A risk assessment analysis conducted for meat consumption and corrected for the aggregated exposure revealed no risk for the consumers

Достижения в области биомониторинга фосфорорганических и хлорорганических пестицидов

Organophosphorus (OPs) and organochlorine (OCs) pesticides are synthetic organic compounds used mainly in agriculture to manage pest problems. Human is occupationally and environmentally exposed to pesticides via inhalation, ingestion and skin absorption. When the pollutant enters human body it is rapidly metabolized to more toxic compounds than the parent compound. Their ability to interfere with biochemical and metabolic pathways in humans makes them compounds of high concern. Acute and chronic exposure has been linked with neurological, cardiovascular, birth effects and male infertility. Human exposure to pesticides is estimated by measuring their metabolites in biological tissues and fluids such as blood, urine, amniotic fluid and hair. In this review study advances on biomonitoring of pesticides are presented. Measured levels of pesticides in urine matrices, amniotic fluid, meconium and hair are presented in order to monitor the human burden and biomonitoring data between occupationally and environmentally exposed populations are being comparedФосфорорганические и хлорорганические пестициды – это синтетические органические соединения, используемые преимущественно в сельском хозяйстве для борьбы с вредителями. Люди подвергаются воздействию пестицидов в процессе профессиональной деятельности и при контакте с зараженной средой через их вдыхание, потребление с пищей и поглощение кожей. При попадании в организм человека поллютант быстро метаболизируется до более токсичных соединений, чем исходное вещество. Их способность взаимодействовать с биохимическими и метаболическими путями в человеческом организме делает их потенциально опасными соединениями. Обнаружена связь между острым и хроническим воздействием пестицидов и неврологическими и кардиоваскулярными эффектами, врожденными отклонениями и мужским бесплодием. Оценка воздействия пестицидов на организм человека проводится путем измерения их метаболитов в биологических тканях и жидкостях, таких как кровь, моча, амниотическая жидкость, а также в волосах. В данном обзоре представлены достижения в области биомониторинга пестицидов. Для оценки нагрузки на человека приведены измеренные уровни пестицидов в моче, амниотической жидкости, меконии и волосах. Проведено сравнение популяций, подвергшихся воздействию пестицидов в связи с производственной деятельностью и при контакте с окружающей средо

Long-term exposure of rabbits to imidaclorpid as quantified in blood induces genotoxic effect

The present in-vivo study focuses on the genotoxic effect of the neonicotinoid pesticide imidacloprid (IMI) in rabbits. The purpose of the study was to establish a possible relationship between exposure to the pesticide (dose and duration) and genotoxicity. Furthermore, an analytical method for the simultaneous determination of IMI and its major metabolite 6-chloronicotinic acid (6-ClNA) in blood was developed and validated. The isolation of the two analytes from blood was performed by liquid-liquid extraction with dichloromethane. Analysis was performed by Liquid Chromatography - Atmospheric Pressure Chemical Ionization - Mass Spectrometry (LC-APCI-MS). The method was applied on the determination of IMI and 6-ClNA in serum samples obtained from rabbits fed with the insecticide at two low doses. Furthermore, parameters of genotoxicity and cytotoxicity were evaluated by measuring binucleated cells with micronuclei (BNMN), micronuclei (MN) and the Cytokinesis Block Proliferation Index (CBPI), in lymphocytes of exposed rabbits. The results revealed a genotoxic effect of IMI for both exposed groups. There were statistically significant differences in the frequencies of BNMN and MN between control and exposed groups but there was no dose-dependence, neither time-dependence of the genotoxic effect for the administered doses. This is the first time that long term exposure to IMI in rabbits was studied for the determination of its genotoxic effect. The genotoxic effect of IMI as it is depicted by the current study is in accordance with previous studies. (C) 2016 Published by Elsevier Ltd