The HBM4EU chromates study - Outcomes and impacts on EU policies and occupational health practices.

Within the EU human biomonitoring initiative (HBM4EU), a targeted, multi-national study on occupational exposure to hexavalent chromium (Cr(VI)) was performed. Cr(VI) is currently regulated in EU under REACH (Registration, Evaluation, Authorisation and Restriction of Chemicals) and under occupational safety and health (OSH) legislation. It has recently been subject to regulatory actions to improve its risk management in European workplaces. Analysis of the data obtained within the HBM4EU chromates study provides support both for the implementation of these regulatory actions and for national enforcement programs and may also contribute to the updating of occupational limit values (OELs) and biological limit values for Cr(VI). It also provides useful insights on the contribution of different risk management measures (RMMs) to further reduce the exposure to Cr(VI) and may support the evaluation of applications for authorisation under REACH. Findings on chrome platers' additional per- and polyfluoroalkyl substances (PFAS) exposure highlight the need to also pay attention to this substance group in the metals sector. A survey performed to evaluate the policy relevance of the HBM4EU chromates study findings supports the usefulness of the study results. According to the responses received from the survey, the HBM4EU chromates study was able to demonstrate the added value of the human biomonitoring (HBM) approach in assessment and management of occupational exposure to Cr(VI). For future occupational studies, we emphasise the need for engagement of policy makers and regulators throughout the whole research process to ensure awareness, relevance and uptake of the results in future policies.

HBM4EU chromates study - the measurement of hexavalent and trivalent chromium in exhaled breath condensate samples from occupationally exposed workers across Europe.

The aim of this study was to investigate the practicability of exhaled breath condensate (EBC) as a biological matrix to detect and measure hexavalent chromium (Cr(VI)) and trivalent chromium (Cr(III)) in workers occupationally exposed to Cr(VI). EBC samples were collected from workers in France, Finland, Italy, The Netherlands and the United Kingdom from three different target activities: chrome platers, stainless steel welders and surface treatment workers. Pre and post working week EBC samples were collected from 177 exposed workers and 98 unexposed workers (control group). Hyphenated chromatography systems with inductively coupled plasma - mass spectrometry (ICP-MS) were for the analysis. The results showed that the occupationally exposed workers had significantly higher levels of Cr(VI) and Cr(III) than the control group. Chrome platers exhibited the highest Cr(VI) levels in their EBC samples, with a significant increase from their pre to post samples for both Cr(VI) and Cr(III). A significant difference was also found between pre and post EBC samples for Cr(III) in welders. This study has shown that EBC has the potential to be a valid, non-invasive biological matrix to assess occupational exposure to Cr(VI) and Cr(III) for biological monitoring assessment, with the ability to detect low level inhalation exposures.

HBM4EU Diisocyanates Study-Research Protocol for a Collaborative European Human Biological Monitoring Study on Occupational Exposure.

Diisocyanates have long been a leading cause of occupational asthma in Europe, and recently, they have been subjected to a restriction under the REACH regulations. As part of the European Human Biomonitoring project (HBM4EU), we present a study protocol designed to assess occupational exposure to diisocyanates in five European countries. The objectives of the study are to assess exposure in a number of sectors that have not been widely reported on in the past (for example, the manufacturing of large vehicles, such as in aerospace; the construction sector, where there are potentially several sources of exposure (e.g., sprayed insulation, floor screeds); the use of MDI-based glues, and the manufacture of spray adhesives or coatings) to test the usability of different biomarkers in the assessment of exposure to diisocyanates and to provide background data for regulatory purposes. The study will collect urine samples (analysed for diisocyanate-derived diamines and acetyl-MDI-lysine), blood samples (analysed for diisocyanate-specific IgE and IgG antibodies, inflammatory markers, and diisocyanate-specific Hb adducts for MDI), and buccal cells (micronucleus analysis) and measure fractional exhaled nitric oxide. In addition, occupational hygiene measurements (air monitoring and skin wipe samples) and questionnaire data will be collected. The protocol is harmonised across the participating countries to enable pooling of data, leading to better and more robust insights and recommendations.

Biological Monitoring: Evidence for Reductions in Occupational Exposure and Risk.

Aims: The aim of this publication is to explore occupational exposure trends from biological monitoring data collected over a period of more than 20 years. The data is stored within the HSE database, which holds more than 950,000 results from 120,000 workers in 8,000 companies. The data were collated for all biological monitoring results for lead, mercury, benzene, and hexamethylene diisocyanate exposures where there have been some regulatory drivers within the reported time period of the data searched. Methods: Relevant results from sample analysed were extracted from the database and categorised by year from 1996 to the end of 2019 for individual blood lead results and individual urine results for mercury, benzene, and hexamethylene diisocyanate. Results were classed by broad occupational sector where possible. Data were reported graphically by analytical biomarker result (as 90th percentile (P90)) and number of samples per year as well as with overall summary statistics. To look at longer-term trends, results were also evaluated as P90 over 6-year periods. Results: In the period 1996-2019, 37,474 blood lead, 11,723 urinary mercury, 9,188 urinary S-phenylmercapturic acid (SPMA, benzene metabolite) and 21,955 urinary hexamethylene diamine (HDA, metabolite of hexamethylene diisocyanate, HDI) samples were analysed and reported. Over the time period the blood lead concentrations saw the P90 reduce from 53 µg/dl 1996) to 24 µg/dl in 2019; the P90 urinary mercury levels reduced from 13.7 µmol/mol creatinine to 2.1 µmol/mol creatinine and the P90 urinary SPMA levels reduced from 133.7 µmol/mol creatinine to 1.7 µmol/mol creatinine. For HDI the P90 results reduced from 2 µmol HDA/mol creatinine in 1996-2000 to 0.7 in 2005-2010 but levels have since increased to 1.0 µmol HDA/mol creatinine (2016-2019). Conclusion: There is strong evidence of reductions in exposure of GB workers to lead, benzene and mercury from the data presented here. These reductions may reflect the impact of national, regional and global regulatory action to reduce exposures however, the loss of high exposure industries (from either GB as a whole or just this dataset i.e., samples are being sent elsewhere) and the increase in automation or substitution also need to be considered as potential factors. The results for HDI show that whilst interventions can reduce exposures significantly, such initiatives may need to be refreshed at intervals to maintain the reductions in exposure. We have observed that exposures move between sectors over time. Waste and recycling (lead, mercury) and tunnelling through contaminated land (benzene) were sectors or tasks associated with significant exposures and may be increasingly areas of concern.

HBM4EU Chromates Study: Determinants of Exposure to Hexavalent Chromium in Plating, Welding and Other Occupational Settings.

Work-related exposures in industrial processing of chromate (chrome plating, surface treatment and welding) raise concern regarding the health risk of hexavalent chromium (Cr(VI)). In this study, performed under the HBM4EU project, we focused on better understanding the determinants of exposure and recognising how risk management measures (RMMs) contribute to a reduction in exposure. HBM and occupational hygiene data were collected from 399 workers and 203 controls recruited in nine European countries. Urinary total chromium (U-Cr), personal inhalable and respirable dust of Cr and Cr(VI) and Cr from hand wipes were collected. Data on the RMMs were collected by questionnaires. We studied the association between different exposure parameters and the use of RMMs. The relationship between exposure by inhalation and U-Cr in different worker groups was analysed using regression analysis and found a strong association. Automatisation of Cr electroplating dipping explained lower exposure levels in platers. The use of personal protective equipment resulted in lower U-Cr levels in welding, bath plating and painting. An effect of wearing gloves was observed in machining. An effect of local exhaust ventilation and training was observed in welding. Regression analyses showed that in platers, exposure to air level of 5 µg/m3 corresponds to U-Cr level of 7 µg/g creatinine. In welders, the same inhalation exposure resulted in lower U-Cr levels reflecting toxicokinetic differences of different chromium species.

HBM4EU chromates study - Overall results and recommendations for the biomonitoring of occupational exposure to hexavalent chromium.

Exposure to hexavalent chromium [Cr(VI)] may occur in several occupational activities, e.g., welding, Cr(VI) electroplating and other surface treatment processes. The aim of this study was to provide EU relevant data on occupational Cr(VI) exposure to support the regulatory risk assessment and decision-making. In addition, the capability and validity of different biomarkers for the assessment of Cr(VI) exposure were evaluated. The study involved nine European countries and involved 399 workers in different industry sectors with exposures to Cr(VI) such as welding, bath plating, applying or removing paint and other tasks. We also studied 203 controls to establish a background in workers with no direct exposure to Cr(VI). We applied a cross-sectional study design and used chromium in urine as the primary biomonitoring method for Cr(VI) exposure. Additionally, we studied the use of red blood cells (RBC) and exhaled breath condensate (EBC) for biomonitoring of exposure to Cr(VI). Personal measurements were used to study exposure to inhalable and respirable Cr(VI) by personal air sampling. Dermal exposure was studied by taking hand wipe samples. The highest internal exposures were observed in the use of Cr(VI) in electrolytic bath plating. In stainless steel welding the internal Cr exposure was clearly lower when compared to plating activities. We observed a high correlation between chromium urinary levels and air Cr(VI) or dermal total Cr exposure. Urinary chromium showed its value as a first approach for the assessment of total, internal exposure. Correlations between urinary chromium and Cr(VI) in EBC and Cr in RBC were low, probably due to differences in kinetics and indicating that these biomonitoring approaches may not be interchangeable but rather complementary. This study showed that occupational biomonitoring studies can be conducted successfully by multi-national collaboration and provide relevant information to support policy actions aiming to reduce occupational exposure to chemicals.

HBM4EU Occupational Biomonitoring Study on e-Waste-Study Protocol.

Workers involved in the processing of electronic waste (e-waste) are potentially exposed to toxic chemicals. If exposure occurs, this may result in uptake and potential adverse health effects. Thus, exposure surveillance is an important requirement for health risk management and prevention of occupational disease. Human biomonitoring by measurement of specific biomarkers in body fluids is considered as an effective method of exposure surveillance. The aim of this study is to investigate the internal exposure of workers processing e-waste using a human biomonitoring approach, which will stimulate improved work practices and contribute to raising awareness of potential hazards. This exploratory study in occupational exposures in e-waste processing is part of the European Human Biomonitoring Initiative (HBM4EU). Here we present a study protocol using a cross sectional survey design to study worker's exposures and compare these to the exposure of subjects preferably employed in the same company but with no known exposure to industrial recycling of e-waste. The present study protocol will be applied in six to eight European countries to ensure standardised data collection. The target population size is 300 exposed and 150 controls. Biomarkers of exposure for the following chemicals will be used: chromium, cadmium and lead in blood and urine; brominated flame retardants and polychlorobiphenyls in blood; mercury, organophosphate flame retardants and phthalates in urine, and chromium, cadmium, lead and mercury in hair. In addition, the following effect biomarkers will be studied: micronuclei, epigenetic, oxidative stress, inflammatory markers and telomere length in blood and metabolomics in urine. Occupational hygiene sampling methods (airborne and settled dust, silicon wristbands and handwipes) and contextual information will be collected to facilitate the interpretation of the biomarker results and discuss exposure mitigating interventions to further reduce exposures if needed. This study protocol can be adapted to future European-wide occupational studies.

HBM4EU chromates study - Reflection and lessons learnt from designing and undertaking a collaborative European biomonitoring study on occupational exposure to hexavalent chromium.

The EU human biomonitoring initiative, HBM4EU, aims to co-ordinate and advance human biomonitoring (HBM) across Europe. As part of HBM4EU, we presented a protocol for a multicentre study to characterize occupational exposure to hexavalent chromium (Cr(VI)) in nine European countries (HBM4EU chromates study). This study intended to collect data on current occupational exposure and to test new indicators for chromium (Cr) biomonitoring (Cr(VI) in exhaled breath condensate and Cr in red blood cells), in addition to traditional urinary total Cr analyses. Also, data from occupational hygiene samples and biomarkers of early biological effects, including genetic and epigenetic effects, was obtained, complementing the biomonitoring information. Data collection and analysis was completed, with the project findings being made separately available. As HBM4EU prepares to embark on further European wide biomonitoring studies, we considered it important to reflect on the experiences gained through our harmonised approach. Several practical aspects are highlighted for improvement in future studies, e.g., more thorough/earlier training on the implementation of standard operating procedures for field researchers, training on the use of the data entry template, as well as improved company communications. The HBM4EU chromates study team considered that the study had successfully demonstrated the feasibility of conducting a harmonised multicentre investigation able to achieve the research aims and objectives. This was largely attributable to the engaged multidisciplinary network, committed to deliver clearly understood goals. Such networks take time and investment to develop, but are priceless in terms of their ability to deliver and facilitate knowledge sharing and collaboration.

Setting up a collaborative European human biological monitoring study on occupational exposure to hexavalent chromium.

The EU human biomonitoring initiative, HBM4EU, aims to co-ordinate and advance human biomonitoring (HBM) across Europe. Within its remit, the project is gathering new, policy relevant, EU-wide data on occupational exposure to relevant priority chemicals and developing new approaches for occupational biomonitoring. In this manuscript, the hexavalent chromium [Cr(VI)] study design is presented as the first example of this HBM4EU approach. This study involves eight European countries and plans to recruit 400 workers performing Cr(VI) surface treatment e.g. electroplating or stainless steel welding activities. The aim is to collect new data on current occupational exposure to Cr(VI) in Europe and to test new methods for Cr biomonitoring, specifically the analysis of Cr(VI) in exhaled breath condensate (EBC) and Cr in red blood cells (RBC) in addition to traditional urinary total Cr analyses. Furthermore, exposure data will be complemented with early biological effects data, including genetic and epigenetic effects. Personal air samples and wipe samples are collected in parallel to help informing the biomonitoring results. We present standard operational procedures (SOPs) to support the harmonized methodologies for the collection of occupational hygiene and HBM samples in different countries.

The Investigation of Unexpected Arsenic Compounds Observed in Routine Biological Monitoring Urinary Speciation Analysis.

This study investigates the identity of two unexpected arsenic species found separately in a number of urine samples sent to the Health and Safety Executive's Health and Safety Laboratory for arsenic speciation (arsenobetaine, AB; arsenite, As3+; arsenate, As5+; monomethylarsonic acid, MMA5+; and dimethylarsinic acid, DMA5+). Micro liquid chromatography coupled to inductively coupled plasma mass spectrometry (µLC-ICP-MS) and electrospray time of flight tandem mass spectrometry (ESI-QqTOF-MS/MS) were used to identify the two arsenic peaks by comparison to several characterized arsenicals: arsenocholine, AC; trimethyl arsine oxide, TMAO; dimethylarsenoacetate, DMAA; dimethylarsenoethanol, DMAE; thio-dimethylarsinate, thio-DMA; thio-dimethylarsenoacetate, thio-DMAA and thio-dimethylarsenoethanol, thio-DMAE. The results from both the ICP-MS and ESI-QqTOF-MS/MS investigations indicate that the unexpected arsenic species termed peak 1 was thio-DMA. While the unexpected arsenic species termed peak 2 has yet to be identified, this investigation shows that it was not AC, TMAO, DMAA, DMAE, thio-DMA, thio-DMAA or thio-DMAE. This study demonstrates the incidence of unexpected arsenic species in both routine and non-routine urine samples from both workers and hospital patients.

Exhaled Breath Condensate: A Novel Matrix for Biological Monitoring to Assess Occupational Exposure to Respirable Crystalline Silica.

Biological monitoring (BM) is a useful way of determining overall exposures to chemical substances; however, in the case of respirable crystalline silica (RCS), this has not been analytically feasible in conventional biological matrices. The aim of this study was to investigate the utility of exhaled breath condensate (EBC) as a potential biological matrix in which to determine exposure to RCS. A small pilot study was undertaken collecting EBC from six quarry workers and six occupationally unexposed persons; the samples were analysed using both single particle inductively coupled plasma mass spectrometry (spICP-MS) and transmission electron microscopy (TEM). The results showed that EBC obtained from the occupationally unexposed persons exhibited low background levels of dissolved silica whilst silica particles of various sizes were present in samples from quarry workers. This is the first study to report EBC as a potential biological matrix that allows differentiation of RCS concentrations between samples from workers and occupationally unexposed controls. The results shown here confirm the presence of RCS in EBC by both spICP-MS and TEM. However, there are difficult analytical challenges still to be overcome before this can be used as a BM method to determine workplace exposure, these are currently being investigated.

The simultaneous detection of trivalent & hexavalent chromium in exhaled breath condensate: A feasibility study comparing workers and controls.

The analytical method outlined in this feasibility study has been used to show that trivalent chromium (Cr(III)) and hexavalent chromium (Cr(VI)) can be detected and measured in exhaled breath condensate (EBC) samples. EBC samples and urine samples were collected from a cohort of 58 workers occupationally exposed to hexavalent chromium compounds and 22 unexposed volunteers (control group). Levels of Cr(III) and Cr(VI) were determined in EBC samples and total chromium levels were determined in urine samples. Pre and post working week samples for both EBC and urine were collected in tandem. Total chromium in urine samples was analysed by inductively coupled plasma mass spectrometry (ICP-MS). Analysis of Cr(III) and Cr(VI) in EBC samples used a hyphenated micro liquid chromatography (µLC) system coupled to an ICP-MS. Separation was achieved using an anion exchange micro-sized column. The results showed that the occupationally exposed workers had significantly higher levels of Cr(III) and Cr(VI) in their EBC samples than the control group, as well as higher levels of total chromium in their urine samples. However, for the exposed workers no significant difference was found between pre and post working week EBC samples for either Cr(III) or Cr(VI). This study has established that Cr(III) and Cr(VI) can simultaneously be detected and measured in 'real' EBC samples and will help in understanding inhalation exposure.

Homicidal arsenic poisoning.

The case of a 50-year-old man who died mysteriously after being admitted to hospital is reported. He had raised the possibility of being poisoned prior to his death. A Coroner's post-mortem did not reveal the cause of death but this was subsequently established by post-mortem trace element analysis of liver, urine, blood and hair all of which revealed very high arsenic concentrations.

Determination of 61 elements in urine samples collected from a non-occupationally exposed UK adult population.

BACKGROUND: levels for 61 elements were established in urine samples collected from 132 occupationally unexposed UK adults. In this study all elements were determined by inductively coupled plasma-mass spectrometry, but methods were 'tailored' to the elements; in total six analytical methods were undertaken. For the first time in a UK population 95th percentile values are reported for 19 elements for which there is no available comparison. Repeat urine samples were collected from some individuals and mixed effects modelling was carried out on the data to give an estimation of variation both between individuals and within the same individual. The mixed effects modelling was undertaken on 31 of the 61 elements for which there were more than two thirds of data above the LOQ and variations of between and within individuals are reported. The analysis found that creatinine adjustment of analyte concentrations was found to be beneficial for 22 of the 31 elements and that smokers were found to exhibit significantly higher cadmium but lower boron than non-smokers. For most elements, the data compare well with other published data but higher concentrations were observed in this study for urinary lead, chromium, vanadium and tungsten.

µLC-ICP-MS determinations of unexposed UK urinary arsenic speciation reference values.

This study provides background levels for five arsenic species in urine, based on urinary data obtained from 95 nonoccupationally exposed volunteers based in the UK. Using a novel, sensitive, robust and reliable speciation methodology, five species of arsenic (arsenobetaine [AB], arsenite [As(3+)], arsenate [As(5+)], monomethylarsonic acid [MMA(5+)] and dimethylarsinic acid [DMA(5+)]) were determined in urine samples collected from 95 adults. The analytical instrumentation used to analyze the urine samples was a hyphenated micro liquid chromatography (µLC) system coupled to an inductively coupled plasma mass spectrometry (ICP-MS). Separation was achieved using an anion exchange micro-sized column. The results presented give the 95th percentile of concentrations, both uncorrected for creatinine (µg/L) and creatinine corrected (µmol/mol) in urine for the 95 volunteers. Statistical analysis was performed on the dataset using a Bayesian model to determine and quantify effects of gender, smoking and diet. The statistical results show that the consumption of fish, shellfish and red wine has a significant elevating effect on AB, DMA and MMA urinary concentrations; however, no significant effect was observed for smoking. The regression model results indicate that creatinine correction was effective for arsenic species As(3+), MMA, DMA and AB. The background levels established here can be used as reference values to help aid interpretation of arsenic speciation results and better assess exposure.

Arsenic speciation in clinical samples: urine analysis using fast micro-liquid chromatography ICP-MS.

Arsenic speciation is a subject that is developing all the time both from improvements in analytical techniques and from increases in toxicological understanding. Despite speciation methods being widely developed, arsenic speciation is not routinely offered as an analysis in clinical laboratory. The work in this paper describes a simple routine method for arsenic speciation that could be easily implemented in clinical laboratories. The method described, a new, fast analytical method for arsenic speciation, is reported using micro-liquid chromatography hyphenated to an inductively coupled plasma mass spectrometer (µLC-ICP-MS). The method uses a low-pressure delivery six-port valve with a 5 cm anion exchange column, which allows a fully resolved separation of five arsenic species (arsenobetaine [AB], arsenite [As(3+)], arsenate [As(5+)], mono-methylarsonic acid [MMA(5+)] and dimethylarsinic acid [DMA(5+)]) in urine in just 6 min. This fast analytical method offers an arsenic speciation method that is feasible for a laboratory that does not have the capability for a dedicated arsenic speciation LC-ICP-MS instrument. The micro-LC system is small, easy to install and is fully integrated with the ICP-MS software. The results reported here are from urine samples from 65 workers in a semiconductor work providing a sample for their routine biological monitoring to assess workplace exposure. Control samples from 20 unexposed people were also determined. Results show that the semiconductor workers exhibit very low levels of arsenic in their urine samples, similar to the levels in the controls, and thus are not significantly exposed to arsenic. Care must be taken when interpreting urinary arsenic species results because it is not always possible to differentiate between dietary and other external sources of exposure.

Beryllium in urine by ICP-MS: a comparison of low level exposed workers and unexposed persons.

PURPOSE: To develop a sensitive and reproducible method for urinary beryllium and to use this method to establish levels in workers at an aluminium smelter and in unexposed persons. METHODS: A method was developed for urinary beryllium using a Thermo ICP-MS Series 1, which was used to determine beryllium concentrations in urine from 62 people with no known occupational exposure to beryllium and 167 workers with potential exposure to beryllium at an aluminium smelter, where beryllium exists as an impurity in the bauxite ore. RESULTS: The analytical method has a detection limit (based on three times the background equivalent concentration of the blank) for beryllium in urine of 6 ng/L. The mean and 90th percentiles of urinary beryllium for workers were 19.5 and 42.0 ng/L and compared with 11.6 and 20.0 ng/L in people not occupationally exposed to beryllium. Statistical analysis using mixed effects models showed that workers had 47% (in 135 paired samples) higher levels of urinary beryllium at the end of the working week compared to the start of week and that the workers who smoked also had significantly higher levels of urinary beryllium compared to those that did not smoke. There was also a statistically significant difference between workers and controls in urinary beryllium concentrations not corrected for creatinine. CONCLUSIONS: A sensitive and reliable analytical method was developed for urinary beryllium by ICP-MS. The workers in this study were exposed to beryllium at very low levels.

Arsenic mobility and speciation in a contaminated urban soil are affected by different methods of green waste compost application.

Application of green waste compost (GWC) to brownfield land is now common practice in soil restoration. However, previous studies have demonstrated both beneficial and detrimental effects on arsenic and metal mobility. In this paper, trace element behaviour was investigated following GWC application, either as surface mulch to, or mixed into soil from a previously described brownfield site in the U.K. Significant differences in arsenic mobility were observed between treatments. Mulching caused most disturbance, significantly increasing soil pore water As, together with Fe, P, Cr, Ni and dissolved organic carbon, the latter was a critical factor enhancing As mobilization. Arsenate was the main inorganic As species in soil pore water, increasing in concentration over time. An initial flush of potentially more toxic arsenite decreased 4 weeks after compost application. Biological processes appeared to play an important role in influencing As mobility. The results point to the necessity for careful management of As-contaminated soils.