The possible contribution of some selected serum micronutrients (beta-carotene, vitamins B12 and C, folic acid and alpha-tocopherol) to spontaneous chromosomal damage was investigated in human peripheral blood lymphocytes from 33 non-smoking healthy donors by the cytokinesis-block micronucleus assay. Labelling of micronuclei with antikinetochore serum was used to discriminate between kinetochore-positive and -negative micronuclei and thus between micronuclei which arise from whole chromosome loss and those which arise from chromosome breaks. Simple correlation analysis showed that age was significantly associated with the increased frequency of micronucleated cells, and this age-related increase in these cells was due to the increase in cells with both kinetochore-positive and -negative micronuclei. Serum micronutrient levels had no apparent significant effects on incidence of micronucleated cells except for the weak positive correlation between vitamin B12 levels and frequency of kinetochore-positive micronucleated cells. Multiple regression analysis with age and serum micronutrient levels as independent variables showed that (a) age was the most influential variable for the frequency of micronucleated cells, (b) the serum vitamin C level was associated with increased frequency of spontaneous micronucleated cells, and this increase was mainly due to the increase in cells with kinetochore-positive micronuclei, and (c) the serum folic acid level was significantly and negatively related to the frequencies of cells with both kinetochore-positive and -negative micronuclei. To avoid the predominant age-effect, we also performed separate multiple regression analysis with age-adjusted frequency of micronucleated cells as dependent variable. The results from this analysis again showed a significant and positive effect of serum vitamin C level on age-adjusted frequency of kinetochore-positive micronucleated cells, while marginal negative effect of folic acid on age-adjusted frequency of total micronucleated cells (P < 0.06) and kinetochore-positive micronucleated cells (P < 0.051) was detected. These results suggest that age and serum vitamin C are definitely variables for frequencies of spontaneous chromosome loss, and that serum folic acid is perhaps another important micronutrient which influence the frequency of spontaneous chromosomal damage.