Human biomonitoring has become a pivotal tool for supporting chemicals' policies. It provides information on real-life human exposures and is increasingly used to prioritize chemicals of health concern and to evaluate the success of chemical policies. Europe has launched the ambitious REACH program in 2007 to improve the protection of human health and the environment. In October 2020 the EU commission published its new chemicals strategy for sustainability towards a toxic-free environment. The European Parliament called upon the commission to collect human biomonitoring data to support chemical's risk assessment and risk management. This manuscript describes the organization of the first HBM4EU-aligned studies that obtain comparable human biomonitoring (HBM) data of European citizens to monitor their internal exposure to environmental chemicals. The HBM4EU-aligned studies build on existing HBM capacity in Europe by aligning national or regional HBM studies. The HBM4EU-aligned studies focus on three age groups: children, teenagers, and adults. The participants are recruited between 2014 and 2021 in 11 to 12 primary sampling units that are geographically distributed across Europe. Urine samples are collected in all age groups, and blood samples are collected in children and teenagers. Auxiliary information on socio-demographics, lifestyle, health status, environment, and diet is collected using questionnaires. In total, biological samples from 3137 children aged 6-12 years are collected for the analysis of biomarkers for phthalates, HEXAMOLL® DINCH, and flame retardants. Samples from 2950 teenagers aged 12-18 years are collected for the analysis of biomarkers for phthalates, Hexamoll® DINCH, and per- and polyfluoroalkyl substances (PFASs), and samples from 3522 adults aged 20-39 years are collected for the analysis of cadmium, bisphenols, and metabolites of polyaromatic hydrocarbons (PAHs). The children's group consists of 50.4% boys and 49.5% girls, of which 44.1% live in cities, 29.0% live in towns/suburbs, and 26.8% live in rural areas. The teenagers' group includes 50.6% girls and 49.4% boys, with 37.7% of residents in cities, 31.2% in towns/suburbs, and 30.2% in rural areas. The adult group consists of 52.6% women and 47.4% men, 71.9% live in cities, 14.2% in towns/suburbs, and only 13.4% live in rural areas. The study population approaches the characteristics of the general European population based on age-matched EUROSTAT EU-28, 2017 data; however, individuals who obtained no to lower educational level (ISCED 0-2) are underrepresented. The data on internal human exposure to priority chemicals from this unique cohort will provide a baseline for Europe's strategy towards a non-toxic environment and challenges and recommendations to improve the sampling frame for future EU-wide HBM surveys are discussed.

Centro Nacional de Sanidad Ambiental Instituto de Salud Carlos 3 28029 Madrid Spain

Croatian Institute of Public Health Division for Environmental Health 1000 Zagreb Croatia

Cyprus International Institute for Environmental and Public Health Cyprus University of Technology 3603 Limassol Cyprus

Cyprus State General Laboratory Ministry of Health P O Box 28648 2081 Nicosia Cyprus

Department Health Protection Laboratoire National de Santé 3555 Dudelange Luxembourg

Department of Biomedical Sciences University of Antwerp 2020 Antwerp Belgium

Department of Clinical Pharmacology Pharmacy and Environmental Medicine University of Southern Denmark 5000 Odense Denmark

Department of Clinical Research University of Basel 4051 Basel Switzerland

Department of Environmental Sciences Jožef Stefan Institute 1000 Ljubljana Slovenia

Department of Epidemiology and Public Health Swiss Tropical and Public Health Institute 4051 Basel Switzerland

Department of Growth and Reproduction Copenhagen University Hospital Rigshospitalet 2100 Copenhagen Denmark

Department of Health Security Finnish Institute for Health and Welfare P O Box 95 70701 Kuopio Finland

Department of Medicine DAME University of Udine Via Colugna 50 33100 Udine Italy

Department of Precision Health Luxembourg Institute of Health 1445 Strassen Luxembourg

Department of Public Health and Welfare Finnish Institute for Health and Welfare P O Box 30 00271 Helsinki Finland

Department of Social Medicine and Epidemiology Faculty of Medicine University of Rijeka 51000 Rijeka Croatia

Department of Sociology University of Antwerp 2020 Antwerp Belgium

Division for Climate and Environmental Health Norwegian Institute of Public Health 0213 Oslo Norway

Environmental Engineering Laboratory Department of Chemical Engineering Aristotle University of Thessaloniki 54124 Thessaloniki Greece

Environmental Health Engineering Institute of Advanced Study Palazzo del Broletto Piazza della Vittoria 15 27100 Pavia Italy

Faculty of Food Science and Nutrition University of Iceland 102 Reykjavik Iceland

Faculty of Public Health Slovak Medical University 833 03 Bratislava Slovakia

German Environment Agency 14195 Berlin Germany

HERACLES Research Center on the Exposome and Health Center for Interdisciplinary Research and Innovation Balkan Center Bldg B 10th km Thessaloniki Thermi Road 57001 Thessaloniki Greece

Institute for Maternal and Child Health IRCCS Burlo Garofolo 34137 Trieste Italy

Institute for Risk Assessment Sciences Utrecht University 3508 TC Utrecht The Netherlands

Masaryk University Research Centre for Toxic Compounds in the Environment 625 00 Bohunice Czech Republic

National Institute for Public Health and the Environment 3721 MA Bilthoven The Netherlands

National Institute of Health 1649 016 Lisbon Portugal

National Institute of Public Health 1000 Ljubljana Slovenia

National Public Health Center 1097 Budapest Hungary

Nofer Institute of Occupational Medicine 91 348 Lodz Poland

Provincial Institute for Hygiene 2000 Antwerp Belgium

Public Health Research Centre NOVA National School of Public Health Universidade NOVA de Lisboa 1099 085 Lisbon Portugal

Santé Publique France Environmental and Occupational Health Division 94415 Saint Maurice France

Swedish Food Agency 751 26 Uppsala Sweden

UK Health Security Agency London SE1 8UG UK

VITO Health Flemish Institute for Technological Research 2400 Mol Belgium

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