The German Environment Agency supports the cooperation on human biomonitoring between the Federal Ministry for the Environment and the German chemical industrial association (VCI). The cooperation is significant for the development of new analytical methods to enable the detection of chemicals in urine or blood samples to which the population may have increased exposure or are of special relevance
The German Ministry for the Environment, Nature Conservation and Nuclear Safety (BMU) and the German Chemical Industry Association (VCI) initiated collaboration in 2010 to provide funding for human biomonitoring (HBM). The main objective of the cooperation is to improve the knowledge about newer chemicals to which the population might be increasingly exposed or which might have an effect on health. Therefore analytical methods are developed which can demonstrate a potential exposure of the general population. Main aim is to develop sensitive methods which are able to detect the smallest concentrations of these substances which occur in the unexposed general public (known as background concentration).
One of the main objectives of the cooperation between the BMU and the VCI is to select up to 50 substances within ten years (substance prioritisation) which meet the criteria of consumer relevance, special relevance to health, or the fact that there is no specific HBM method available. This is where work begins on the development of specific methods of analysis and their application in suitable population surveys. The VCI is responsible for the development of methods, and the German Environment Agency (UBA) supports the Federal Environment Ministry (BMU) in the application of the methods in suitable studies. UBA provides administration and management of the cooperation.
The cooperation originally was formed for ten years. The selection of substances was finished in early 2020. The development of the analytical methods is a complex and lengthy process therefore the cooperation has been extended by another five years to 2025.
Committees and members
The BMU and the VCI cooperate in two committees: the HBM Expert Group and the Steering Committee.
The members of the HBM Expert Group have backgrounds in chemical-analytical science, expertise in toxicology, environmental medicine and/ or regulatory affairs and work for the Federal Environment Ministry (BMU), the "Federal Institute for Risk Assessment (BfR)", the "Federal Institute for Occupational Health and Safety (BAuA)" and the German Environment Agency (UBA), in science (universities, private and state institutes), and in the chemical industry.
Membership is contingent upon recommendation and approval by both cooperation partners; membership is – apart from the governmental representatives - personal rather than institutional. The members of the Steering Committee are representatives from the BMU, UBA and VCI (or its membership organisations).
Selection of substances and method development
Until 2019 the HBM Expert Group convened at the beginning and in autumn of every year, firstly to select the substances for which methods are to be developed, and secondly to discuss the individual critical steps of this development. Since 2020 the Expert Group meets only once a year for exchange and discussion on progress made in the development of methods.
First discussions on the selection of substances commenced before the official start of the cooperation and were based on lists compiled by different international organisations (e.g. the list of CMR (carcinogenic, mutagenic, reprotoxic) substances, the Candidate List to REACH Annex XIV, or the list of endocrine disruptors) and also considered substances of concern under discussion at the time. The BMU commissioned the BfR in 2010 to draw up a list of some 120 substances which might be relevant for the cooperation. This list was based on BfR’s assessment work in the frame of consumer health protection and on basis of worldwide on different levels discussed consumer relevant substances (e.g. Candidate List of REACH, and cosmetics ingredients). The basic criteria for selection of substances from the available lists were:
Good to very good bioavailability of the substances
Toxicological relevance
Exclusion of substances for which an HBM method exists, either for the compound itself or for its active components or metabolites (related research in “The MAK Collection for Occupational Health and Safety” and laboratory list of the IPASUM (Institute and Outpatient Clinic of Occupational, Social, and Environmental Medicine), Erlangen). If the methods found seemed not appropriate for this cooperation (e.g. not sensitive enough) an intense study of the provided methods was performed.
Exclusion of substances with low likelihood of exposure (no consumer relevance) (e.g. research in the SPIN database)
The finally compiled list built the bases for further discussions on appropriate substances within the HBM Expert Group. Subsequent meetings have focused on a regular review of international chemical lists. Recommendations of the participating institutions to take up new substances are heard, and toxicological fact sheets are used to determine the substance’s appropriateness for the cooperative project.
Regular items on the agenda of the HBM Expert Group discussion include: aspects of substance toxicity (health relevance), potential of consumer exposure to the substances, and the availability of a specific HBM method for tracing the substance itself or its metabolites (as exposure or effect biomarkers) in human urine or blood samples. In cases where an HBM method exists in the occupational safety field (MAK Collection), the expert group discusses the biomarkers used, possible detection and determination limits and/or the need for an additional metabolism study. The decision-making process increasingly weighs whether the available information from human, animal and cell culture tests enable the HBM Commission to derive an assessment (HBM) value.
The results of the HBM Expert Group’s discussion generate a list of proposed substances, which features about ten chemicals every year and for which there was sufficient data available to classify them as eligible for the cooperation project.
The list of proposed substances from the HBM Expert Group is submitted to the Steering Committee for its annual spring meeting. The Steering Committee selects up to 5 substances per year for which the cooperative partners are to develop HBM methods. Consideration is given to whether the VCI is able to find industrial sponsors for the substance whose expertise is required for the effective development of analysis methods. Selection of the substances was concluded in spring 2020.
After the up to five substances have been chosen by the Steering Committee, the VCI commissions a laboratory (often a member of the HBM Expert Group) to develop a suitable method. The June 2013 issue of BMU-Umwelt features an article with details of the challenges involved in developing these methods.
Selected substances
In the ten years of joint project work on human biomonitoring (2010-2020), the BMU and VCI selected fifty substances for which the first human biomonitoring analysis methods were developed. These substances include phthalates and phthalate substitutes (plasticizers), flame retardants (for furniture or clothes), solvents (in cleaning products or wall colours), cosmetics additives (UV filters, fragrances, preservatives), plastics additives (UV-stabilizers), biocides and allergenic substances (preservatives for food, cosmetics or household products).
Detailed information on the selected substances, their uses, and their chemical names are to be found in the Table 'Selected Substances' below.
More than half of analytical methods already developed
The BMU/VCI cooperation has already developed several urine and, in some cases, blood analysis methods for more than half of the selected substances. They enable the analytical determination of substances for which there had been no method of detection (in these low concentrations) anywhere in the world.
These include a method for the analysis of metabolites of the phthalate DPHP and the phthalate substitutes DINCH, DEHTP, TOTM, DnBA and DEHA. These plasticizers are increasingly used to replace e.g. the now banned DEHP in different fields of application, for example in toys, food packaging or medical tubes. A method has also been developed for the analysis of MDI, an ingredient in single-component foam for the fixing and insulation of window and door frames. Methods to measure the following have been developed for: 2-MBT, which is used as a vulcanisation accelerator in the production of household rubber products and tyres; Lysmeral, 7-Hydroxycitronellal and Geraniol, fragrances frequently used in cosmetics and cleaning products and detergents. UV-filters and preservatives can be found in several cosmetics (like cream and sun blocker) for some of them analytical methods have been developed (4-MBC, Octocrylene, CMI/MI (3:1), EHS, DHHB). Appropriate analytical methods to measure the background level for the flame retardants HBCDD and TDCPP are also available.
More information on the single methods developed can be found in the Table “Developed analysis methods”.
The development of a method for three substances (cyclosiloxanes D4, D5, D6) was discontinued due to difficulties of analysis. Additionally, it was not possible to develop a method for the substance Keromet MD as it degrades to unspecific parts very fast once it has entered the human body.
Once a method has been developed by an internationally renowned laboratory commissioned by the VCI, the method and the established exposure biomarkers is presented in peer reviewed journals and is often also presented at specialist conferences, whereby the method becomes publicly available.
The new analytical methods fulfill the high standards on validity and quality of the working group ‘Analyses in biological Materials’ of the Permanent Senate Commission for the Investigation of Health Hazards of Chemical Compounds in the Work Area (known as the MAK Commission) of the German Research Foundation (DFG). These standards include:
Identification of one or more specific biomarkers (mainly metabolites)
Development and validation of an analytical method for the determination and quantification of the biomarker(s) in urine or blood.
In addition, the new method has to be applied by 40 non-occupationally exposed volunteers to approve the capability of the method to measure the background exposure.
The first application of the methods in population surveys occurs through the German Environment Agency in the analysis of human samples of the Environmental Specimen Bank or samples taken for the German Environmental Survey (GerES), a population representative survey.
For example, the methods for DINCH, DPHP, DEHTP, the solvents NMP and NEP and the methode for 2-MBT have been applied to the German Environmental Survey for Children and Adolescents, GerES 2014-2017 (GerES V) . The samples of the Environmental Specimen Bank have been used to apply the methods for DINCH, DPHP, NMP und NEP, HBCDD, DEHTP and CMI/MI (3:1). Some methods will also be applied in GerES VI (German Environmental Survey on Adults). The results of these studies will be presented in peer reviewed journals or on the respective web pages.
With these results, for example, an estimation of the average corporal burden of the population with these substances is possible. This supports the assessment of the effectiveness of existing European and German legal regulations for the use of these substances – or may contribute to further regulations.
Progress made in the BMU/VCI project was part of the reason why Germany has been asked to coordinate an EU-wide programme on human-biomonitoring with the acronym HBM4EU. Under this initiative, the European Commission supplies a co-fund of 50 million euros between 2017 and 2021 to bring together and advance human-biomonitoring activities in the EU member states and a couple of associated countries. The German Environment Agency (UBA) is leading and successfully managing this complex project and is also incorporating the results of the BMU/VCI cooperation into this work.
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Denghel H, Göen T.. Determination of the UV absorber 2-(2H-benzotriazol-2-yl)-4,6-di-tert-pentylphenol (UV 328) and its oxidative metabolites in human urine by dispersive liquid-liquid microextraction and GC–MS/MS, Journal of Chromatography B, Volume 1144, 2020, 122071, https://doi.org/10.1016/j.jchromb.2020.122071
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Höllerer C, Becker G, Göen T, Eckert E. Regioselective ester cleavage of di-(2-ethylhexyl) trimellitates by porcine liver esterase. Toxicology In Vitro, March 2018; 47:178-185. https://doi.org/10.1016/j.tiv.2017.11.015
Höllerer C, Müller J, Göen T, Eckert E. Isomeric separation and quantitation of di-(2-ethylhexyl) trimellitates and mono-(2-ethylhexyl) trimellitates in blood by LC–MS/MS. Journal of Chromatography B 1 September 2017; Volumes 1061-1062:153-162. https://doi.org/10.1016/j.jchromb.2017.07.014
Jäger T, Bäcker S, Brodbeck T, Leibold E, Bader M. Quantitative determination of urinary metabolites of geraniol by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Analytical Methods. online-first published Nov 2 2020. https://doi.org/10.1039/D0AY01582B
Kasper-Sonnenberg M, Koch HM, Apel P, Rüther M, Pälmke C, Brüning T, Kolossa-Gehring M. Time trend of exposure to the phthalate plasticizer substitute DINCH in Germany from 1999 to 2017: Biomonitoring data on young adults from the Environmental Specimen Bank (ESB). Int. J. Hyg. Environ. Health, Volume 222, Issue 8, 2019, pp 1084-1092, https://doi.org/10.1016/j.ijheh.2019.07.011
Krystek P, Beeltje H, Noteboom M, van den Hoeven EM, Houtzager MMG. Analytical human biomonitoring method for the identification and quantification of the metabolite BDCPP originated from the organophosphate flame retardant TDCPP in urine, Journal of Pharmaceutical and Biomedical Analysis, Volume 170, 2019, pp 169-175, https://doi.org/10.1016/j.jpba.2019.03.036
Lessmann F, Kolossa-Gehring M, Apel P, Rüther M, Pälmke C, Harth V, Brüning T, Koch HM. German Environmental Specimen Bank: 24-hour urine samples from 1999 to 2017 reveal rapid increase in exposure to the para-phthalate plasticizer di(2-ethylhexyl) terephthalate (DEHTP), Environment International, Volume 132, 2019, https://doi.org/10.1016/j.envint.2019.105102
Lessmann F, Schütze A, Weiss T, Brüning T, Koch HM. Determination of metabolites of di(2-ethylhexyl) terephthalate (DEHTP) in human urine by HPLC-MS/MS with on-line clean-up, Journal of Chromatography B, 2016, Volume 1011:196-203. http://dx.doi.org/10.1016/j.jchromb.2015.12.042
Murawski A, Fiedler N, Schmied-Tobies, MIH, Rucic E, Schwedler G, Stoeckelhuber M, Scherer G, Pluym N, Scherer M, Kolossa-Gehring, M. Metabolites of the fragrance 2-(4-tert-butylbenzyl)propionaldehyde (lysmeral) in urine of children and adolescents in Germany – Human biomonitoring results of the German Environmental Survey 2014–2017 (GerES V). International Journal of Hygiene and Environmental Health, 229, 2020, 113594. https://doi.org/10.1016/j.ijheh.2020.113594
Murawski A, Schmied-Tobies MIH, Rucic E, Schmidtkunz C, Küpper K, Leng G, Eckert E, Kuhlmann L, Göen T, Daniels A, Schwedler G, Kolossa-Gehring, M. Metabolites of 4-methylbenzylidene camphor (4-MBC), butylated hydroxytoluene (BHT), and tris(2-ethylhexyl) trimellitate (TOTM) in urine of children and adolescents in Germany – human biomonitoring results of the German Environmental Survey GerES V (2014–2017), Environmental Research, 2020, 110345, ISSN 0013-9351, https://doi.org/10.1016/j.envres.2020.110345.
Murawski A, Schmied-Tobies MIH, Rucic E, Schettgen T, Bertram J, Conrad A, Kolossa-Gehring M. (2020). The methylisothiazolinone and methylchloroisothiazolinone metabolite N-methylmalonamic acid (NMMA) in urine of children and adolescents in Germany – Human biomonitoring results of the German Environmental Survey 2014–2017 (GerES V). International Journal of Hygiene and Environmental Health, 227, 2020, 113511. https://doi.org/10.1016/j.ijheh.2020.113511
Murawski A, Schmied-Tobies MIH, Schwedler G, Rucic E, Gries W, Schmidtkunz C, Küpper K, Leng G, Conrad A, Kolossa-Gehring, M. 2-Mercaptobenzothiazole in urine of children and adolescents in Germany – Human biomonitoring results of the German Environmental Survey 2014–2017 (GerES V). International Journal of Hygiene and Environmental Health, 228, 2020, 113540. https://doi.org/10.1016/j.ijheh.2020.113540
Nehring A, Bury D, Kling H-W, Weiss T, Brüning T, Koch HM. Determination of human urinary metabolites of the plasticizer di(2-ethylhexyl) adipate (DEHA) by online-SPE-HPLC-MS/MS, Journal of Chromatography B, Volume 1124, 2019, pp 239-246, https://doi.org/10.1016/j.jchromb.2019.06.019
Nehring A, Bury D, Ringbeck B, Kling H-W, Otter R, Weiss T, Brüning T, Koch HM. Metabolism and urinary excretion kinetics of di(2-ethylhexyl) adipate (DEHA) in four human volunteers after a single oral dose, Toxicology Letters, Volume 321, 2020, Pages 95-102, https://doi.org/10.1016/j.toxlet.2019.12.006
Pluym N, Petreanu W, Weber T, Scherer G, Scherer M, Kolossa-Gehring M. Biomonitoring data on young adults from the Environmental Specimen Bank suggest a decrease in the exposure to the fragrance chemical 7-hydroxycitronellal in Germany from 2000 to 2018. International Journal of Hygiene and Environmental Health 227, 2020 https://doi.org/10.1016/j.ijheh.2020.113508
Ringbeck B, Bury D, Hayen H, Weiss T, Brünung T, Koch HM. Determination of di-n-butyl adipate (DnBA) metabolites as possible biomarkers of exposure in human urine by online-SPE-LC-MS/MS. Journal Of Chromatography B, Volume 1141, 2020, 122029. https://doi.org/10.1016/j.jchromb.2020.122029
Scherer M, Petreanu W, Weber T, Scherer G, Pluym N, Kolossa-Gehring M. Human biomonitoring in urine samples from the Environmental Specimen Bank reveals a decreasing trend over time in the exposure to the fragrance chemical lysmeral from 2000 to 2018. Chemosphere, Volume 265, 2021, 128955, ISSN 0045-6535, https://doi.org/10.1016/j.chemosphere.2020.128955.
Schettgen T, Rüther M, Weber T, Kraus T, Kolossa-Gehring M. N-methylmalonamic acid (NMMA) as metabolite of methylisothiazolinone and methylchloroisothiazolinone in 24-h urine samples of the German Environmental Specimen Bank from 2000 to 2017 – exposure and time trends, Chemosphere. 2020. https://doi.org/10.1016/j.chemosphere.2019.125743
Schettgen T, Bertram J, Kraus T. Quantification of N-methylmalonamic acid in urine as metabolite of the biocides methylisothiazolinone and chloromethylisothiazolinone using gas chromatography-tandem mass spectrometry. Journal Of Chromatography B, Volumes 1044–1045, 15 February 2017, Pages 185-193. https://doi.org/10.1016/j.jchromb.2017.01.019
Schettgen T, Kraus T. Urinary excretion kinetics of the metabolite N-methylmalonamic acid (NMMA) after oral dosage of chloromethylisothiazolinone and methylisothiazolinone in human volunteers. Archives of Toxicology. December 2017, Volume 91, Issue 12, pp 3835–3841. http://dx.doi.org/10.1007/s00204-017-2051-5
Schmidtkunz C, Küpper K, Weber T, Leng G, Kolossa-Gehring M. A biomonitoring study assessing the exposure of young German adults to butylated hydroxytoluene (BHT). International Journal of Hygiene and Environmental Health 228 (2020) https://doi.org/10.1016/j.ijheh.2020.113541
Schmidtkunz C, Gries W, Weber T, Leng G, Kolossa-Gehring M. Internal exposure of young German adults to di(2-propylheptyl) phthalate (DPHP): Trends in 24-h urine samples from the German Environmental Specimen Bank 1999–2017, International Journal of Hygiene and Environmental Health, Volume 222, Issue 3, 2019, pp 419-424, https://doi.org/10.1016/j.ijheh.2018.12.008
Schmied-Tobies MIH, Murawski A, Rucic E, Schwedler G, Bury D, Kasper-Sonnenberg M, Koslitz S, Koch HM, Brüning T, Kolossa-Gehring M. Alkyl pyrrolidone solvents N–methyl–2–pyrrolidone (NMP) and N–ethyl–2–pyrrolidone (NEP) in urine of children and adolescents in Germany – human biomonitoring results of the German Environmental Survey 2014–2017 (GerES V), Environment International, Volume 146, 2021, 106221 https://doi.org/10.1016/j.envint.2020.106221
Schwedler G, Conrad A, Rucic E, Koch HM, Leng G, Schulz C, Schmied-Tobies MIH, Kolossa-Gehring M. Hexamoll® DINCH and DPHP metabolites in urine of children and adolescents in Germany. Human biomonitoring results of the German Environmental Survey GerES V, 2014–2017. International Journal of Hygiene and Environmental Health, Volume 229, 2020, 113397. https://doi.org/10.1016/j.ijheh.2019.09.004
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Stoeckelhuber M, Scherer M, Peschel O, Leibold E, Bracher F, Scherer G, Pluym N. Human metabolism and urinary excretion kinetics of the UV filter Uvinul A plus® after a single oral or dermal dosage. International Journal of Hygiene and Environmental Health, 227, 2020, 113509, https://doi.org/10.1016/j.ijheh.2020.113509
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Stoeckelhuber M, Krnac D, Pluym N, Scherer M, Leibold E, Scherer G. A validated UPLC–MS/MS method for biomonitoring the exposure to the fragrance 7-hydroxycitronellal. Journal Of Chromatography B 15 November 2017;Volumes 1068-1069:261-267. https://doi.org/10.1016/j.jchromb.2017.10.040
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Ulrich N, Bury D, Koch HM, Rüther M, Weber T, Käfferlein H-U, Weiss T, Brüning T, Kolossa‑Gehring M. Metabolites of the alkyl pyrrolidone solvents NMP and NEP in 24-h urine samples of the German Environmental Specimen Bank from 1991 to 2014, International Archives of Occupational and Environmental Health. https://doi.org/10.1007/s00420-018-1347-y
The UBA’s motto, For our environment (“Für Mensch und Umwelt”), sums up our mission pretty well, we feel. In this video we give an insight into our work.
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