RESUMO
Technical grade branched nonylphenol (NP) was determined in human urine by online solid phase extraction-ultra high-performance liquid chromatography-tandem mass spectrometry (SPE-UHPLC-MS/MS). Prior to analysis, urine specimens were simply diluted and enzymatically deconjugated. The run time of the chromatography, including SPE and re-equilibration, was 9 min per injection. The enzymatic cleavage of NP conjugates was optimised with incurred sample material from a human metabolism study: the highest recoveries were obtained with ß-glucuronidase from E. coli K 12 in 0.1 M ammonium acetate at pH 6.5, within a minimal hydrolysis time of 30 to 60 min. Using sodium acetate instead of ammonium acetate led to systematically decreased recovery rates. The analytical method was validated regarding its precision (coefficients of variation: 2.9-7.4%), accuracy (relative recovery rates: 93-105%), robustness (relative recovery rates in individual urine matrices: 92-117%), selectivity, and limit of quantification (1.0 µg L-1). Fundamental aspects in the analysis of technical product mixtures such as NP, comprising various isomers and homologues, were considered. Validation results, an exposure scenario and the application of the procedure to real samples, show that it enables a rugged monitoring of NP exposures above, at, and significantly below health-based guidance values, corresponding to daily NP intakes in the low µg kg-1 d-1 range. On the other hand, background levels in non-specifically exposed populations cannot be detected with this method. Hence, while alternative approaches should be pursued for NP analysis at environmental trace level, the speed and simplicity of our method are ideal for high-throughput human biomonitoring in occupational medicine.
Assuntos
Escherichia coli , Espectrometria de Massas em Tandem , Humanos , Espectrometria de Massas em Tandem/métodos , Cromatografia Líquida de Alta Pressão , Extração em Fase Sólida/métodosRESUMO
Nonylphenol (NP) is an endocrine-disrupting anthropogenic chemical that is ubiquitous in the environment. Human biomonitoring data and knowledge on internal NP exposure are still sparse, and its human metabolism is largely unknown. Therefore, in this study, we investigated human metabolism and urinary excretion of NP. Three male volunteers received a single oral dose of 1 mg 13C6-labeled NP (10.6-11.7 µg/kg body weight). Consecutive full urine voids were collected for 48 h. A metabolite screening identified nine ring- and/or side chain-oxidized metabolites. We chose the most promising hits, the alkyl chain-oxidized metabolites hydroxy-NP (OH-NP) and oxo-NP, for quantitative investigation next to the parent NP. For this purpose, we newly synthesized specific n - 1-oxidized monoisomeric analytical standards. Quantification of the polyisomeric metabolites was performed via online-solid phase extraction-LC-MS/MS with stable isotope dilution using a previously published consensus method. Alkyl chain hydroxylation (OH-NP) constituted the major metabolism pathway representing 43.7 or 62.2% (depending on the mass transition used for quantification) of the NP dose excreted in urine. The urinary excretion fraction (FUE) for oxo-NP was 6.0 or 9.3%. The parent NP, quantified via an analogous isomeric 13C6-NP standard, represented 6.6%. All target analytes were excreted predominately as glucuronic acid conjugates. Excretion was rather quick, with concentration maxima in urine 2.3-3.4 h after dosing and biphasic elimination kinetics (elimination half-times first phase: 1.0-1.5 h and second phase: 5.2-6.8 h). Due to its high FUE and insusceptibility to external contamination (contrary to parent NP), OH-NP represents a robust and sensitive novel exposure biomarker for NP. The novel FUEs enable to robustly back-calculate the overall NP intakes from urinary metabolite levels in population samples for a well-informed cumulative exposure and risk assessment.
Assuntos
Fenóis/metabolismo , Fenóis/urina , Administração Oral , Adulto , Cromatografia Líquida , Voluntários Saudáveis , Humanos , Cinética , Masculino , Pessoa de Meia-Idade , Estrutura Molecular , Fenóis/administração & dosagem , Espectrometria de Massas em TandemRESUMO
2-Mercaptobenzothiazole (MBT) is one of the most important vulcanization accelerators in the industrial production of rubber, especially car tires. Given its wide use in household articles and industrial rubber products it has a high potential to migrate into the environment. Humans can be exposed by dermal, oral, or inhalative routes. Incorporated MBT is excreted in urine, mainly as conjugates to glucuronide, sulfate, and mercapturic acid. On the basis of these facts MBT has been selected as a substance of high interest in the large scale 10-year German project on human biomonitoring (HBM); a cooperation between the German Federal Ministry for the Environment (BMUB) and the German Chemical Industry Association (VCI) with the objective of developing new analytical methods for relevant chemicals. The presented method was developed to determine MBT in human urine to reliably investigate the internal human MBT dose. Total MBT is measured after enzymatic hydrolysis followed by application of high-pressure liquid chromatography tandem mass spectrometry (HPLC-MS-MS) in positive-electrospray-ionization mode (ESI+) using isotope-dilution quantification. High sample throughput could be obtained by use of the column-switching technique. Optimization yielded an analytical method with a low and reproducible limit of detection (LOD) of 0.4 µg L(-1) and a limit of quantification (LOQ) of 1 µg L(-1), and low relative standard deviations in the range 1.6-5.8 %. A small biomonitoring study covering unexposed humans and occupationally exposed workers was performed to establish the feasibility and reliability of the method. MBT was found in only one urine sample from the unexposed humans, at a value of 10.8 µg MBT per liter, whereas it was found in all samples from the tested workers at values of up to 6210 µg MBT per liter.
Assuntos
Benzotiazóis/urina , Cromatografia Líquida de Alta Pressão/métodos , Exposição Ambiental/análise , Espectrometria de Massas em Tandem/métodos , Calibragem , Humanos , Hidrólise , Limite de Detecção , Exposição Ocupacional/análise , Reprodutibilidade dos Testes , Borracha/química , Sensibilidade e Especificidade , Espectrometria de Massas por Ionização por Electrospray/métodosRESUMO
4,4'-Methylene diphenyl diisocyanate (MDI) is one of the most important isocyanates in the industrial production of polyurethane and other MDI-based synthetics. Because of its high reactivity, it is known as a sensitizing agent, caused by protein adducts. Analysis of MDI is routinely done by determination of the nonspecific 4,4'-methylenedianiline as a marker for MDI exposure in urine and blood. Since several publications have reported specific adducts of MDI and albumin or hemoglobin, more information about their existence in humans is necessary. Specific adducts of MDI and hemoglobin were only reported in rats after high-dose MDI inhalation. The aim of this investigation was to detect the hemoglobin adduct 5-isopropyl-3-[4-(4-aminobenzyl)phenyl]hydantoin (ABP-Val-Hyd) in human blood for the first time. We found values up to 5.2 ng ABP-Val-Hyd/g globin (16 pmol/g) in blood samples of workers exposed to MDI. Because there was no information available about possible amounts of this specific MDI marker, the analytical method focused on optimal sensitivity and selectivity. Using gas chromatography-high-resolution mass spectrometry with negative chemical ionization, we achieved a detection limit of 0.02 ng ABP-Val-Hyd/g globin (0.062 pmol/g). The robustness of the method was confirmed by relative standard deviations between 3.0 and 9.8 %. Combined with a linear detection range up to 10 ng ABP-Val-Hyd/g globin (31 pmol/g), the enhanced precision parameter demonstrates that the method described is optimized for screening studies of the human population.
Assuntos
Poluentes Ocupacionais do Ar/química , Indústria Química , Hemoglobinas/química , Hidantoínas/sangue , Isocianatos/química , Exposição Ocupacional , Poluentes Ocupacionais do Ar/sangue , Poluentes Ocupacionais do Ar/urina , Biomarcadores/sangue , Biomarcadores/urina , Calibragem , Cromatografia Gasosa-Espectrometria de Massas/métodos , Hemoglobinas/metabolismo , Humanos , Hidantoínas/urina , Isocianatos/sangue , Isocianatos/urina , Limite de Detecção , Reprodutibilidade dos TestesRESUMO
A pilot study was conducted for human biomonitoring of the suspected carcinogen 2-chloroprene. For this purpose, urine samples of 14 individuals occupationally exposed to 2-chloroprene (exposed group) and of 30 individuals without occupational exposure to alkylating substances (control group) were analysed for six potential mercapturic acids of 2-chloroprene: 4-chloro-3-oxobutyl mercapturic acid (Cl-MA-I), 4-chloro-3-hydroxybutyl mercapturic acid (Cl-MA-II), 3-chloro-2-hydroxy-3-butenyl mercapturic acid (Cl-MA-III), 4-hydroxy-3-oxobutyl mercapturic acid (HOBMA), 3,4-dihydroxybutyl mercapturic acid (DHBMA) and 2-hydroxy-3-butenyl mercapturic acid (MHBMA). In direct comparison with the control group, elevated levels of the mercapturic acids Cl-MA-III, MHBMA, HOBMA and DHBMA were found in the urine samples of the exposed group. Cl-MA-I and Cl-MA-II were not detected in any of the samples, whereas HOBMA and DHBMA were found in all analysed urine samples. Thus, for the first time, it was possible to detect HOBMA and Cl-MA-III in human urine. The mercapturic acid Cl-MA-III could be confirmed as a specific metabolite of 2-chloroprene in humans providing evidence for the intermediate formation of a reactive epoxide during biotransformation. The main metabolite, however, was found to be DHBMA showing a distinct and significant correlation with the urinary Cl-MA-III levels in the exposed group. The obtained results give new scientific insight into the course of biotransformation of 2-chloroprene in humans.
Assuntos
Acetilcisteína/urina , Alquilantes/metabolismo , Cloropreno/metabolismo , Exposição Ocupacional , Adulto , Alquilantes/efeitos adversos , Biomarcadores/urina , Biotransformação , Estudos de Casos e Controles , Cloropreno/efeitos adversos , Cromatografia Líquida , Monitoramento Ambiental/métodos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Exposição Ocupacional/efeitos adversos , Projetos Piloto , Medição de Risco , Espectrometria de Massas em Tandem , Regulação para Cima , Adulto JovemRESUMO
Climbazole is a preservative and an anti-dandruff ingredient with applications in various cosmetic products. The general population is therefore exposed to this chemical, and exposure monitoring is desirable. We have postulated a pathway for the human metabolism of climbazole, leading to two specific metabolites which can be excreted via urine. An analytical method for the determination of these metabolites in human urine was developed and validated. The sample preparation includes an enzymatic hydrolysis protocol. The measurement as such is based on online solid phase extraction (SPE), coupled to ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Intra- and inter-series coefficients of variation (CV) were determined in the concentration range from 1 µg/l to 100 µg/l with spiked pooled urine samples, and they were consistently below 15%, mostly below 10%. The corresponding accuracies (mean relative recovery rates) in spiked pooled urine varied from 97% to 103%. The robustness of the method was estimated by spiking individual urine samples. At 1 µg/l, the robustness was rather limited due to interfering matrix peaks in several samples, but excellent results were obtained at 10 µg/l and 100 µg/l, with CVs between 7% and 14% and accuracies from 101% to 110%. Matrix interferences often seemed to be associated with higher creatinine contents (≥2.0 g/l) of the samples. We subsequently applied the method to urine specimens from a human metabolism study involving documented climbazole exposures. We were able to identify and quantify the postulated metabolites in those real samples, thus validating our metabolism hypothesis. We also investigated the precision and accuracy of the enzymatic deconjugation with the real samples. The deconjugation step was found to be highly repeatable and largely quantitative. Both metabolites formed glucuronides, though varying fractions were also excreted in unconjugated (free) forms. Phase II conjugates other than glucuronides did not seem to be produced in significant amounts. With our method, both climbazole metabolites can be reliably quantified in the range between about 1.5 µg/l (depending on matrix interferences in individual samples) and at least 500 µg/l.
RESUMO
2-Mercaptobenzothiazole (2-MBT) is widely used as a vulcanisation accelerator and is contained in many products made from natural rubber, e.g. car tires. Additionally, it is used as a fungicide in paint or fibre. Systemically human exposure to 2-MBT can occur via dermal and oral uptake or inhalation. Locally, 2-MBT can cause skin sensitisation. The International Agency for Research on Cancer (IARC) classified 2-MBT as probably carcinogenic to humans. 516 urine samples of 3- to 17-year-old children and adolescents living in Germany were analysed for the concentration of 2-MBT in the population representative German Environmental Survey for Children and Adolescents 2014-2017 (GerES V). 2-MBT was quantified above the limit of quantification (LOQ) of 1.0 µg/L in 50% of the 516 samples analysed. The geometric mean of urinary 2-MBT concentration was 1.018 µg/L and 0.892 µg/gcreatinine, the arithmetic mean was 1.576 µg/L (1.351 µg/gcrea). The median concentration was below the LOQ. Analyses of subgroups revealed higher 2-MBT concentrations in children aged 3-5 years compared to 14- to 17-year-old adolescents. All urinary 2-MBT concentrations were well below the health-based guidance value HBM-I for children of 4.5 µg/L. Therefore, current exposure levels are - according to current knowledge - not of concern. For the first time, reference values can be derived for 2-MBT for children and adolescents in Germany. This will facilitate to recognise changing exposure levels in this population group in Germany and identification of unusually high exposures.
Assuntos
Benzotiazóis/urina , Poluentes Ambientais/urina , Adolescente , Monitoramento Biológico , Criança , Pré-Escolar , Feminino , Alemanha , Humanos , Masculino , Inquéritos e QuestionáriosRESUMO
Di(2-propylheptyl) phthalate (DPHP) is used as a substitute for high molecular weight phthalates like di(2-ethylhexyl) phthalate (DEHP) which were subjected to authorization under REACh in 2015. An earlier study on the time trend of exposure in human 24-h urine samples from the German Environmental Specimen Bank has revealed that metabolites of DPHP emerged in 2009 and 2012 (Schütze et al., 2015). In order to better assess a potential trend and the present state of exposure to DPHP, we now measured 180 urine samples from the German Environmental Specimen Bank, 60 per year, collected in 2011, 2014 and 2017, randomized and blinded before analysis. Together with the previously analyzed samples, data for a total of 480 samples covering 19 years from 1999 to 2017 was thus generated. We were able to show that DPHP exposure of the studied population, university students from Münster (Northwestern Germany), has remained essentially constant since 2011, after a rapid increase starting around 2009. Even so, urinary metabolite concentrations were always in the low ppb or sub-ppb range, indicating that DPHP exposure of the general population is substantially lower than for other modern plasticizers, and far below levels currently regarded as critical. DPHP is a plasticizer which is mostly used in non-sensitive applications with little probability of close contact to humans. Still, we observed how temporal trends of DPHP exposure largely follow trends of DPHP consumption in the Western European market. Our results hence demonstrate the potential of biomonitoring to sensitively detect the effects of industrial product strategy on the environment, even when biomarkers are present only at trace level.
Assuntos
Ácidos Ftálicos/urina , Plastificantes/análise , Adulto , Monitoramento Biológico/estatística & dados numéricos , Bancos de Espécimes Biológicos , Feminino , Alemanha , Humanos , Masculino , Adulto JovemRESUMO
In this publication the challenges to cope for the aim to obtain innovative biomonitoring methods in our laboratory are visualized for di(2-propylheptyl)phthalate, 2-mercaptobenzothiazole, 3,5-di-tert-butyl-4-hydroxytoluene, 4-nonylphenol, 4-tert-octylphenol, 3-(4-methylbenzylidene)camphor, 4,4'-methylene diphenyl diisocyanate, and Hexabromocyclododecane. For these substances new specific markers were explored based on animal or human kinetic data with urine being the preferred matrix compared to blood. The determination of these markers was complex in all cases, because the sample preparation as well as the detection by high performance liquid chromatography, capillary gas chromatography coupled to tandem mass spectrometers or high resolution mass spectrometry should enable the lowest possible detection limit by use of minimal biological sample volumes. To get a first hint of a possible background level, the analytical methods were applied to urine samples of about 40 persons for each chemical. For Di(2-propylheptyl)phthalate and 2-Mercaptobenzothiazole first results are presented from population biomonitoring.
Assuntos
Monitoramento Ambiental/métodos , Benzotiazóis/urina , Cânfora/análogos & derivados , Cânfora/urina , Poluentes Ambientais/sangue , Poluentes Ambientais/urina , Humanos , Hidrocarbonetos Bromados/urina , Isocianatos/sangue , Fenóis/urina , Ácidos Ftálicos/urinaRESUMO
Pyrethrum as well as synthetic pyrethroids like allethrin, resmethrin, phenothrin, tetramethrin, cyfluthrin, cypermethrin, deltamethrin or permethrin are among the insecticides most often used worldwide. With a sensitive and valid gas-chromatographic-high resolution mass spectrometric method, it is possible to detect all pyrethrum and pyrethroid metabolites in one analytical run. Thus, for the first time a background level of trans-chrysanthemumdicarboxylic acid (CDCA) in urine (95th percentile: 0.15 microg/l) as a characteristic, e.g. for a pyrethrum exposure was found. Following a pyrethrum exposure lasting 1 day, CDCA was found in 27 out of 30 subjects with concentrations going up to 54 microg/l urine (mean: 1.1+/-4.4 microg/l). To obtain information about the elimination kinetics of pyrethrum in humans, urinary excretion of CDCA was investigated in three volunteers after oral intake of 0.3mg pyrethrin I. CDCA was detected during the first 36 h after intake with elimination being most rapid during the first 4h (mean elimination half-life: 4.2h).
Assuntos
Inseticidas/urina , Piretrinas/urina , Administração Oral , Biomarcadores/urina , Chrysanthemum cinerariifolium , Monitoramento Ambiental , Humanos , Masculino , Piretrinas/administração & dosagem , Piretrinas/farmacocinética , Valores de ReferênciaRESUMO
Persistent organic pollutants are widespread in the environment, and are associated with a particular health and ecological concern. The human body burden of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDDs/Fs), polybrominated dibenzo-p-dioxins and dibenzofurans (PBDDs/Fs), polychlorinated biphenyls (PCBs), polybrominated diphenylether (PBDEs), and hexabromocyclododecanes (HBCDs) was determined. Blood samples were collected in Germany, originating from 42 randomly selected subjects between 20 and 68 years old. The median (95th percentile) concentrations, expressed as WHO2005-TEQ for PCDD/PCDFs and dioxin-like PCBs, were 6.2 (19.1) pg/g l.w. and 4.1 (8.8) pg/g l.w., respectively. PBDDs/Fs were found with a median of 2.8 pgTEQ/g l.w. and a 95th percentile of 8.7 pgTEQ/g l.w. (using similar interim TEF values as for PCDDs/Fs) On a median basis, the contribution of PCDD/Fs, dioxin-like PCBs, and PBDDs/Fs to total TEQ were 47%, 31%, and 21%, respectively. The sum of the 6 non-dioxin-like PCBs exhibited a median of 267ng/g l.w. and a 95th percentile of 834ng/g l.w. The median value for the sum of six tetra- to hepta-PBDE congeners was 1.7ng/g l.w. (95th percentile: 4.9ng/g l.w.). BDE 209 was the most abundant congener with a median of 1.8ng/g l.w. HBCDs were only found in some samples, and concentrations ranged between the limit of detection (5ng/g l.w.) and the limit of quantification (16ng/g l.w.). Results for PBDEs and HBCDs are comparable to other European studies. Our study demonstrated that the body burden of PCDD/Fs and PCBs declined continously since the last three decades, but exposure may exceed precautionary guideline levels.
Assuntos
Dibenzofuranos/sangue , Dioxinas/sangue , Poluentes Ambientais/sangue , Retardadores de Chama/análise , Éteres Difenil Halogenados/sangue , Hidrocarbonetos Halogenados/sangue , Adulto , Idoso , Monitoramento Ambiental , Feminino , Alemanha , Humanos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
A new developed gas chromatographic-high resolution mass spectrometric method for the sensitive simultaneous determination of trans-chrysanthemumdicarboxylic acid, cis- and trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane carboxylic acid, cis-3-(2,2-dibromovinyl)-2,2-dimethylcyclopropane carboxylic acid, 3-phenoxybenzoic acid and 4-fluoro-3-phenoxybenzoic acid in human urine is presented. These metabolites are biomarkers for an exposure to pyrethrum, allethrin, resmethrin, phenothrin, tetramethrin, cyfluthrin, cypermethrin, deltamethrin or permethrin. Therefore, with the help of this method for the first time a complete assessment of exposure to pyrethroid and pyrethrin insecticides is possible. After acid hydrolysis and extraction with tert-butyl-methyl-ether the residue is derivatized with 1,1,1,3,3,3-hexafluoroisopropanol and analyzed by GC/HRMS in electron impact mode (detection limits < 0.1 microg/l) as well as in negative chemical ionization mode (detection limit < 0.05 microg/l urine).
Assuntos
Cromatografia Gasosa-Espectrometria de Massas/métodos , Piretrinas/urina , Humanos , Padrões de Referência , Sensibilidade e EspecificidadeRESUMO
UNLABELLED: For the purpose of implementing steam sterilization of 2-[18F]FDG (FDG) in the final container into routine production, we have validated and established a fully automated dispensing and sterilization system, thereby considerably reducing the radiation burden to the personnel. METHODS: The commercially available system combines aseptic dispensing of the product solution under a miniaturized laminar flow unit with subsequent steam sterilization, realized by heating of the product in the final containers by an autoclave included in the dispensing unit, thus incorporating current pharmaceutical manufacturing standards for the production of parental radiopharmaceuticals. The efficiency of the used sterilization cycle, the stability of FDG under the conditions of sterilization and the stability of the final product towards radiolysis was investigated with respect to various pH-formulations. RESULTS: The system was found to be fully valid for filling of vials in a laminar flow class A (US-class 100) environment and for sterilization of FDG in the final container. The pH for sterilizing FDG solutions must be slightly acidic to avoid decomposition. A pH of 5.5 appears to be optimal and gives FDG of very high radiochemical purity (approximately 99%). In addition, radiolysis of FDG in solutions of high activity concentration was significantly lower at pH 5.5 than at neutral pH. CONCLUSION: Terminal sterilization enables the production of FDG in full compliance with GMP-regulations even in Class C or D (US class 10,000 or 100,000) laboratories.
Assuntos
Fluordesoxiglucose F18/administração & dosagem , Compostos Radiofarmacêuticos/administração & dosagem , Esterilização/métodos , Fluordesoxiglucose F18/normas , Humanos , Concentração de Íons de Hidrogênio , Tomografia por Emissão de Pósitrons/métodos , Tomografia por Emissão de Pósitrons/normas , Compostos Radiofarmacêuticos/normas , Vapor , Esterilização/instrumentaçãoRESUMO
The aim of the presented investigation was to document challenges encountered during implementation and qualification of a method for bisphenol A (BPA) analysis and to develop and discuss precautions taken to avoid and to monitor contamination with BPA during sample handling and analysis. Previously developed and published HPLC-MS/MS methods for the determination of unconjugated BPA (Markham et al. Journal of Analytical Toxicology, 34 (2010) 293-303) [17] and total BPA (Markham et al. Journal of Analytical Toxicology, 38 (2014) 194-203) [20] in human urine were combined and transferred into another laboratory. The initial method for unconjugated BPA was developed and evaluated in two independent laboratories simultaneously. The second method for total BPA was developed and evaluated in one of these laboratories to conserve resources. Accurate analysis of BPA at sub-ppb levels is a challenging task as BPA is a widely used material and is ubiquitous in the environment at trace concentrations. Propensity for contamination of biological samples with BPA is reported in the literature during sample collection, storage, and/or analysis. Contamination by trace levels of BPA is so pervasive that even with extraordinary care, it is difficult to completely exclude the introduction of BPA into biological samples and, consequently, contamination might have an impact on BPA biomonitoring data. The applied UPLC-MS/MS method was calibrated from 0.05 to 25ng/ml. The limit of quantification was 0.1ng/ml for unconjugated BPA and 0.2ng/ml for total BPA, respectively, in human urine. Finally, the method was applied to urine samples derived from 20 volunteers. Overall, BPA can be analyzed in human urine with acceptable recovery and repeatability if sufficient measures are taken to avoid contamination throughout the procedure from sample collection until UPLC-MS/MS analysis.
Assuntos
Compostos Benzidrílicos/química , Compostos Benzidrílicos/urina , Cromatografia Líquida de Alta Pressão/métodos , Fenóis/química , Fenóis/urina , Extração em Fase Sólida/métodos , Espectrometria de Massas em Tandem/métodos , Compostos Benzidrílicos/isolamento & purificação , Humanos , Modelos Lineares , Fenóis/isolamento & purificação , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
The analytical determination of hemoglobin adducts was used as an effective biomonitoring tool after a fire outbrake at a chemical plant close to Cologne/Germany in 2008. More than 1000 people (e.g. fire-men, police officers, and workers) were potentially exposed to acrylonitrile and ethylene. Air monitoring in the surrounding was performed, and acrylonitrile was measured in concentrations up to 20 ppm, the mean value being 7 ppm (time range: 8 h). As many people were concerned about their individual body burden, biomonitoring was recommended for all people involved. 816 persons took advantage of this opportunity and came for blood sampling to the occupational health department of our company. Regarding the lifespan of erythrocytes up to 3 months, it was possible to analyze hemoglobin adducts of acrylonitrile and ethylene during and after the accident. In case of acrylonitrile the hemoglobin adduct N-(2-cyanoethyl) valine and regarding ethylene, N-(2-hydroxyethyl) valine was determined. As a result, the body burden was in nearly all cases within our internal adduct reference values (CyEtVal<15 µg/L blood or <612 pmol/g globin; HyEtVal<15 µg/L blood or 646 pmol/g globin). In about 1% of the cases, the adduct concentrations were slightly above these reference values. This means that the body burden measured by biomonitoring turned out to be far lower than the one expected from the air data. Therefore, following chemical incidents, in case biomonitoring is meaningful, it is highly recommended beside of air monitoring.
Assuntos
Acrilonitrila/sangue , Vazamento de Resíduos Químicos , Monitoramento Ambiental/métodos , Etilenos/sangue , Exposição Ocupacional/análise , Valina/análogos & derivados , Acidentes de Trabalho , Socorristas , Bombeiros , Alemanha , Hemoglobinas/análise , Humanos , Polícia , Valina/sangueRESUMO
Di(2-propylheptyl) phthalate (DPHP), a high molecular weight phthalate, is primarily used as a plasticizer in polyvinyl chloride and vinyl chloride copolymers for technical applications, as a substitute for other phthalates currently being scrutinized because of endocrine disrupting effects. We determined urinary excretion fractions of three specific DPHP metabolites (mono-2-(propyl-6-hydroxy-heptyl)-phthalate (OH-MPHP), mono-2-(propyl-6-oxoheptyl)-phthalate (oxo-MPHP) and mono-2-(propyl-6-carboxy-hexyl)-phthalate (cx-MPHxP)) after oral dosing of five volunteers with 50mg labelled DPHP-d4 and subsequent urine sampling for 48 h. These excretion fractions are used to back calculate external intakes from metabolite measurements in spot urine analysis. Following enzymatic hydrolysis to cleave possible conjugates, we determined these urinary metabolites by HPLC-NESI-MS/MS with limits of quantification (LOQ) between 0.3 and 0.5 µg/l. Maximum urinary concentrations were reached within 3-4h post dose for all three metabolites; elimination half-lives were between 6 and 8h. We identified oxo-MPHP as the major oxidized metabolite in urine representing 13.5±4.0% of the DPHP dose as the mean of the five volunteers within 48 h post dose. 10.7±3.6% of the dose was excreted as OH-DPHP and only 0.48±0.13% as cx-MPHxP. Thus, within 48 h, 24.7±7.6% of the DPHP dose was excreted as these three specific oxidized DPHP metabolites, with the bulk excreted within 24h post dose (22.9±7.3%). These secondary, oxidized metabolites are suitable and specific biomarkers to determine DPHP exposure. In population studies, however, chromatographic separation of these metabolites from other isomeric di-isodecyl phthalate (DIDP) metabolites is warranted (preferably by GC-MS) in order to distinguish DPHP from general DIDP exposure. Palatinol(®), Hexamoll(®) and DINCH(®) are registered trademarks of BASF SE, Germany.
Assuntos
Monitoramento Ambiental/métodos , Ácidos Ftálicos/farmacocinética , Ácidos Ftálicos/urina , Adulto , Biomarcadores , Meia-Vida , Humanos , Masculino , Pessoa de Meia-Idade , Ácidos Ftálicos/químicaRESUMO
This publication describes a method for the determination of total bisphenol A (BPA and conjugated BPA) following enzyme hydrolysis and is intended as a companion to our previously developed analytical method for the determination of free BPA (the aglycone) in human blood and urine using high-performance liquid chromatography-tandem mass spectrometry ( 1). That free BPA method provided a means to account for and/or eliminate background contamination and demonstrated accuracy and reproducibility in both matrices fortified with BPA or a surrogate analyte ((13)C BPA) at a low method quantitation limit (MQL) of 0.1-0.2 ng/mL. In contrast to the free BPA method results and based on stringent accuracy, precision and confirmation criteria set for the MQLs of the method developed for total BPA, the MQL achieved in blood was 1.020-2.550 and 0.510-1.020 ng/mL in urine. These data showed higher MQLs than the desired MQLs of 0.5 ng/mL (blood) and 0.2 ng/mL (urine) with increased variability between analyses which demonstrates the importance of generating method validation data with each analysis. In contrast, the MQL achieved for (13)C BPA-G (monoglucuronide as a surrogate analyte in blood was 0.2-0.5 and 0.2 ng/mL in urine illustrating that the method is capable of meeting lower MQL requirements if the contribution from exogenous BPA can be well controlled. This method for the determination total BPA in human blood and urine is intended to be used in conjunction with the free BPA method ( 1) to obtain accurate and complete BPA biomonitoring data to support human exposure assessments.
Assuntos
Compostos Benzidrílicos , Cromatografia Líquida de Alta Pressão/métodos , Monitoramento Ambiental/métodos , Poluentes Ambientais , Fenóis , Espectrometria de Massas por Ionização por Electrospray/métodos , Espectrometria de Massas em Tandem/métodos , Compostos Benzidrílicos/sangue , Compostos Benzidrílicos/urina , Calibragem , Cromatografia Líquida de Alta Pressão/instrumentação , Monitoramento Ambiental/instrumentação , Poluentes Ambientais/sangue , Poluentes Ambientais/urina , Humanos , Limite de Detecção , Fenóis/sangue , Fenóis/urina , Padrões de Referência , Reprodutibilidade dos Testes , Espectrometria de Massas por Ionização por Electrospray/instrumentação , Espectrometria de Massas em Tandem/instrumentaçãoRESUMO
An analytical method for the determination of ß-hydroxyethoxyacetic acid (HEAA), the main urinary metabolite of 1,4-dioxane was developed and validated. The presented method involves liquid-liquid extraction of HEAA from the urine samples, followed by silylation and subsequent analytical separation and detection using GC-MS. The method is characterized by its simple and fast sample preparation in combination with a robust chromatography. The use of isotope dilution analysis enables an efficient compensation of matrix related effects and analyte losses due to sample workup. The excellent reliability and reproducibility of the method is demonstrated by the good accuracy and precision data. Within-day precision and day-to-day precision ranged from 0.6 to 1.2% and 1.5 to 2.6%, respectively. The mean relative recovery of the method was found to be 98-101%. The LOD and LOQ of HEAA were determined to be 0.2mg/L and 0.6mg/L, respectively. In summary, the presented analytical method is well suited to be used for routine biomonitoring of occupational exposure to 1,4-dioxane.
Assuntos
Acetatos/urina , Cromatografia Gasosa-Espectrometria de Massas/métodos , Acetatos/química , Acetatos/metabolismo , Dioxanos/química , Dioxanos/metabolismo , Humanos , Técnicas de Diluição do Indicador , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Di-(2-propylheptyl)-phthalate (DPHP) is a specific phthalic acid ester of isomeric C10 alcohols. It is classified as high molecular weight phthalate and marketed as plasticizer for polyvinyl chloride (PVC). The increase of its production volume and its wide field of application suggest a possible background exposure of the human population as found for other phthalates, making suitable analytical methods necessary. The aim of the presented analytical report is the sensitive and selective determination of the three major DPHP metabolites mono-2-(propyl-6-hydroxy-heptyl)-phthalate (OH-MPHP), mono-2-(propyl-6-oxoheptyl)-phthalate (oxo-MPHP) and mono-2-(propyl-6-carboxy-hexyl)-phthalate (cx-MPHxP) in human urine. Most of the published analytical methods for phthalate metabolites use high pressure liquid chromatography tandem mass spectrometry (HPLC-MS/MS). The methods presented here allow a comparison of chromatographic separation between HPLC-MS/MS and gas chromatography high resolution mass spectrometry (GC-HRMS), which is useful to distinguish between DPHP and DIDP. The enhanced detection limits range between 0.05-0.1µg/L for GC-HRMS and 0.1-0.2µg/L for HPLC-MS/MS.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Cromatografia Gasosa-Espectrometria de Massas/métodos , Ácidos Ftálicos/urina , Espectrometria de Massas em Tandem/métodos , Adulto , Calibragem , Humanos , Redes e Vias Metabólicas , Ácidos Ftálicos/metabolismo , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
We developed and validated an analytical method for the simultaneous determination of several chlorine and non-chlorine containing mercapturic acids in urine as specific metabolites of the hazardous chemicals 2-chloroprene and epichlorohydrin. The method involves an online column switching arrangement for online solid phase extraction of the analytes with subsequent analytical separation and detection using LC-MS/MS. The developed method enables for the first time the determination of Cl-MA-I (4-chloro-3-oxobutyl mercapturic acid), Cl-MA-II (4-chloro-3-hydroxybutyl mercapturic acid), Cl-MA-III (3-chloro-2-hydroxy-3-butenyl mercapturic acid) and HOBMA (4-hydroxy-3-oxobutyl mercapturic acid) as potential biomarkers of 2-chloroprene in urine. Additionally, CHPMA (3-chloro-2-hydroxypropyl mercapturic acid) as a specific metabolite of epichlorohydrin in urine and DHBMA (3,4-dihydroxybutyl mercapturic acid) can be determined. The analytical method proved to be both sensitive and reliable with detection limits ranging from 1.4 µg/L (for Cl-MA-III) to 4.2 µg/L (for HOBMA). Intra- and interday imprecision was determined to range from 4.7 to 11.8%. Due to the good accuracy and precision and the low limits of detection the developed method is well suited for application in biomonitoring studies in order to determine occupational exposure to 2-chloroprene and epichlorohydrin.