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In the Czech Republic, the Human Biomonitoring Project (CZ-HBM) was launched in 1994 as an integral part of the nationwide Environmental Health Monitoring System (EHMS). Until now, the HBM covers two time periods: the first covered 1994-2003 and the second, 2005-2009. Altogether three population groups were included in the HBM: adults (blood donors aged 18-58 years), children aged 8-10 years, and breastfeeding primiparas. Sampling is organized on a yearly basis. Altogether three groups of biomarkers were analyzed: (a) selected heavy metals (Pb, Cd, Hg) and essential elements (Cu, Se, Zn) in blood and urine of adults and children, (b) indicator PCBs, DDT, DDE, HCB and HCHs in human milk and blood serum of adults and (c) cytogenetic changes in peripheral lymphocytes in blood of adults and children. This paper is focused on the general design of the CZ-HBM, the trends over time, and reference values. A significant downward time trend was observed for the blood lead levels in adults and children. Increased urinary cadmium levels were observed in Czech children compared to German children. The blood and urinary mercury levels were higher in women than in men. The levels of indicator PCB congeners in the Czech human milk samples were still higher than in most other European countries, because of existing hot spots. The levels of organochlorine pesticides showed a substantial continuous downward trend.

The developed method for mercury speciation analysis has been validated and used for the biomonitoring study of mercury species in human hair. Statistical evaluation proved the reliability of simplified determination of inorganic mercury (difference between total mercury and methylmercury). The results of the validation showed that the method is very well suitable for the determination of both species of mercury in hair for biomonitoring purposes. Non-exposed schoolchildren from three areas in the western and central part of the Czech Republic were chosen as the target group. Tenth of a microgram per gram of the total mercury were generally found in the analyzed hair; values higher than 1 microg g(-1) were detected only exceptionally. Comparable results were obtained for two western areas and differed significantly from those for the third area located in the central part of the Czech Republic. In the areas examined, the mean methylmercury contents amounted to 23-46% of the total mercury in the hair. The results confirm an assumption that exposure to mercury does not pose a significant risk to the population in the Czech Republic.

Over a 5-year period, from 1996 to 2001, blood selenium levels were recorded in a set of 2414 healthy blood donors (1781 men and 633 women; 880 smokers and 1534 nonsmokers) living in four selected areas of the Czech Republic. About 100 blood samples per year and region were analyzed using the HGA method. The internal and external quality controls of this method were performed throughout the duration of the study. In general, blood selenium concentrations (81.9 and 106.7 microg/l for median and 90th percentile, respectively) did not reach optimum values; in approximately 10% of the population sample, values lower than 60 microg/l were detected. Nonetheless, the values obtained increased significantly, with median concentrations of 73.2 microg/l in 1996 and 91.5 microg/l in 2001. The percentage of subjects with a whole blood selenium level of less than 60 microg/l also decreased from nearly 20% in 1996 to 0.2% in 2001. No substantial regional or gender-related differences were observed, but significantly lower blood selenium levels were found in smokers as opposed to nonsmokers. Although mild selenium deficiency continues to be observed, the data presented do not indicate extremely low selenium levels in the population sample.

A simple method for the determination of sub-microgram amounts of mercury species in human hair is described. The method is based on the selective leaching of methylmercury from hair with hydrochloric acid followed by determination of the separated mercury species using cold vapour atomic absorption spectrometry. The results obtained by the proposed method are compared with results obtained by the solvent extraction method. Both methods gave the same results and are suitable for the determination of methylmercury and inorganic mercury in hair with natural (10–100 ng g–1) or higher contents of mercury with precision higher than 10%.

The method of Guo et aL (AnaL Chim. Acta, 1997, 349, 313-318) for the determination of the toxicologically relevant arsenic in urine was verified and then used for the determination of arsenic in urine of the Czech population for monitoring purposes. Statistical evaluation at the level alpha = 0.05 did not prove any significant differences between industrial and agricultural regions, between males and females and smokers and nonsmokers. Likewise no differences were found among children in all the regions monitored. In the adult population small differences were found between some regions but these differences were not dependent on industrial pollution. The values of toxicologically relevant arsenic are low for all regions. The summarised value of the median for all groups together is 3.5 microg (g creatinine)(-1).

SUMMARY As the concentration of methylmercury (MeHg) in the environment is insignificant, hair can be used as a suitable matrix to estimate endogenous MeHg exposure. A validated analytical method with AMA 254 spectrometer was used for the determination of inorganic mercury and methylmercury species in the hair of dentists, workers in fish industry and professionally non-exposed adults. ANOVA and QC Expert software was used for statistical evaluation. The number of amalgam fillings in oral cavity, consumption of fish, gender, smoking habits and age of the subjects were taken into account. A significantly higher level of inorganic bound mercury (Hg in ) was found in the hair of dentists. The number of amalgam fillings had a slightly significant effect on Hg in ; fish consumption had a significant influence on MeHg and slightly also on Hg in . Other parameters were not significant.

A sampling procedure appropriate for the determination of mercury in whole blood was tested by using both inactive controls and a 197Hg mercury radio-indicator. To exclude the influence of the instrumental device (an AMA 254 single-purpose mercury atomic absorption spectrometer) on the determination of mercury in whole blood, the function of the instrument was checked by using rat blood with metabolised 197Hg. The measurement procedure was found to be free of errors. However, the study showed that the material used for the sampling vessels is a crucial parameter for obtaining accurate analytical results. The stability of solutions and samples was tested towards polyethylene (PE) and polypropylene (PP) vessels. PE displayed a time-dependent increase in the mercury content both in the samples and in the blood control material. The probable cause of this increase was direct contamination from the material of the vessel and/or diffusion of mercury from the environment through the vessel walls related to a strong complexing affinity of the sample matrix. This assumption was confirmed by supplying the vessels with the complexing agent Na2EDTA (0.05 mol L(-1)). Commercial PP vessels for blood sampling (Sarstedt S-Monovette Metall Analytik) did not give rise to statistically significant variations in mercury content in the samples and blood control material over a 30-day period.

In the framework of the system of monitoring the environmental impact on population health, the concentration of lead, cadmium and selenium in blood and cadmium in urine was measured in adults (n = 670), children (n = 599) and umbilical blood (n = 549) using atomic absorption spectrophotometry. Furthermore, cytogenetic analysis of peripheral lymphocytes in all population groups under study was investigated. The median blood Pb level for the overall group of adults (47.8 micrograms/l, i.e. 0.23 mumol/1) was significantly higher in men (51.5 micrograms/l, i.e. 0.25 mumol/l). Smoking significantly influenced the blood Pb level in women. The 90th percentile in no group exceeded the value of 100 micrograms/l (0.48 mumol/l). The median blood Cd level in adults (0.9 microgram/l, i.e. 0.008 mumol/l) depends on smoking habit (1.25 micrograms/l, i.e. 0.01 mumol/l). The median urine Cd level was 0.585 microgram/g creatinine (0.59 mumol/mole creatinine) in adults and 0.37 microgram/g creatinine (0.37 mumol/mole creatinine) in children. The median blood Se level (53.5 micrograms/l, i.e. 0.68 mumol/l) was found to be higher in the group of non-smokers (57.5 micrograms/l, i.e 0.73 mumol/l). Lead and selenium level were significantly lower in the umbilical blood. Cytogenetic analysis results showed age-dependent average percentages of aberrant cells: 1.1% in umbilical blood, 1.27% in children and 1.71 in adults in line with the reference values for the Czech population.

Energies and intensities of electron transition of different forms of dithizone were calculated by the method of molecular orbitals (LCAO MO SCF LCI) and compared with the measured spectra of dithizone and its ions. It has been found that the spectrum of nondissociated dithizone corresponds to a compound which is close to the structure of the enol form. The spectrum of dithizone in moderate alkaline solutions corresponds to the enol form with the SH- group dissociated and in strongly alkaline solutions (approx. 2N NaOH) to the enol form with both hydrogen atoms dissociated. The spectrum of dithizone in 60% H2SO4 corresponds to the enol form with the imine group protonated.