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The article summarises the results of Czech Human Biomonitoring over a period of three decades. Human biomonitoring in the Czech Republic was commenced at the National Institute of Public Health, Prague, in 1994 as part of the Environmental Health Monitoring System and was later linked to European and worldwide professional activities in this area. During the course of this project the whole range of organic and inorganic xenobiotics and essential substances was monitored in the blood, urine and hair of the adult and child populations, in the breast milk of breastfeeding women and in other, less common matrices. The results have provided valuable long-term time series that show how established preventive measures can work in practice and how they correspond to decreasing population exposure. Results can also be used to verify whether or not, and how quickly, can individuals be exposed to newly used chemical substances. Results of human biological monitoring are important for public health professionals to establish reference values, to provide comparison with health limits, to assess health risks, for health policy purposes and international comparison. Appropriately processed results can serve to inform and educate the public about human population burden by chemical substances in the environment. Long-term experience in the field of human biomonitoring enables experts of the National Institute of Public Health to participate in many foreign projects focused on monitoring the exposure of various population groups to chemical substances from the environment. [ABSTRACT FROM AUTHOR]