OBJECTIVE To investigate the relationship of zinc in the causation of neural tube defects and so explore the possible role as a cause of these defects. METHODS Stored serum samples collected as part of the Medical Research Council's multicentre randomised trial section of vitamin supplementation in the prevention of neural tube defects were retrieved in respect of women who had had an affected pregnancy together with four matched unaffected controls per case. One control was selected from each of the four randomisation groups: 1. no extra vitamins; 2. folic acid supplementation alone; 3. folic acid and other vitamin supplementation; and 4. other vitamin supplementation, such that it was matched with an NTD case by study centre, maternal age (the year of birth differed at most by one year) and duration of storage of the serum sample (in most instances to within the same year). METHODS A prospective case-control study using serum samples stored at -40 degrees C. The serum samples were collected from women at entry to the trial, immediately before the women became pregnant and at about 12 weeks of pregnancy. METHODS Twenty-seven women with a pregnancy associated with a neural tube defect and 108 matched controls with unaffected pregnancies. RESULTS There was no association between serum zinc levels and neural tube defects; the mean values in microgram/dl were: 82 and 85 in cases and controls, respectively, at entry to the study; 86 and 87 micrograms/dl just before pregnancy; and 76 and 80 micrograms/dl in the first trimester of pregnancy. Folic acid supplementation did not have an effect on serum zinc levels. CONCLUSIONS Within the range of serum zinc levels observed among women in our study, serum zinc is not a cause of neural tube defects, and periconceptional folic acid supplements do not compromise the zinc status of the mother or the embryo with respect to zinc intake.