BACKGROUND The influence of maternal vitamin D on pregnancy outcomes, including preterm birth (PTB), is unclear due to different experimental designs and study populations (patient race and sample size) of previous studies. We aimed to investigate the relationship between 25-hydroxyvitamin D (25[OH] D) levels and PTB among pregnant women in southern China. MATERIAL AND METHODS A total of 11 641 pregnant women were retrospectively enrolled between January 2016 and April 2019. Vitamin D concentrations were evaluated by electrochemiluminescence immunoassay. Logistic regression analysis was used to analyze the association between vitamin D and PTB. RESULTS The average 25(OH) D concentration was 59.3±21.5 nmol/L; 34.8% of patients were vitamin D deficient, 43.0% were vitamin D insufficient (25[OH] D <50 nmol/L and 50-74.9 nmol/L, respectively). In total, 3.6% of newborns were born prematurely. Comparing the pre-term and full-term groups, 45.7% versus 42.9% and 29.8% versus 35% were vitamin D deficient and insufficient, respectively These differences were not significant (P>0.05). However, the mean vitamin D status was significantly different between the pre-term and full-term groups (61.3±21.3 and 59.1±21.5 nmol/L, respectively). No association was found between vitamin D deficiency/insufficiency and PTB in unadjusted or adjusted models, compared with vitamin D sufficiency (adjusted odds ratio, 1.016; 95% confidence interval, 0.794-1.301 and 0.842; 0.641-1.106, respectively). CONCLUSIONS Low maternal 25(OH) D levels are common in southern China. However, low vitamin D status in pregnant women appears to be unrelated to PTB. Measuring vitamin D level alone is therefore not sufficient to predict PTB.