OBJECTIVE: The purpose of this study was to investigate the maternal and children's myo-inositol, glucose, and zinc status in association with spina bifida risk. STUDY DESIGN: Sixty-three mothers and 70 children with spina bifida and 102 control mothers and 85 control children were investigated. The maternal and child serum myo-inositol, serum glucose, and red blood cell zinc concentrations were measured when the child was between 1 and 3 years old. These data were compared between cases and control subjects. The association with spina bifida was expressed by the ratio of geometric means and by odds ratios and 95% CI for a cutoff value at the extreme 10th percentile of the control group. RESULTS: The geometric mean of the maternal myo-inositol concentration tended to be 5% (95% CI, -1% to 11%) lower in cases. Interestingly, the odds ratio for the extreme low maternal myo-inositol concentration was 2.6 (95% CI, 1.1-6.0). The glucose and zinc concentrations were significantly higher at 7% (95% CI, 4%-10%) and significantly lower at 5% (95% CI, 0%-9%), in case mothers compared with control mothers. The odds ratios (95% CI) for maternal high glucose and low zinc concentrations were 4.6 (2.0-10.5) and 2.9 (1.2-7.0), respectively. The geometric mean of the myo-inositol concentration tended to be 7% (95% CI, 0%-14%) lower in children with spina bifida; the glucose and zinc concentrations were comparable. CONCLUSION: Maternal myo-inositol, glucose, and zinc status are associated with the risk of spina bifida in offspring. Furthermore, the myo-inositol status of the child seems to contribute to this risk as well.