Folate status for women during early pregnancy has been investigated, but data for women during mid-pregnancy, late pregnancy, or lactation are sparse or lacking. Between May and July 2014, we conducted a cross-sectional study in 1211 pregnant and lactating women from three representative regions in China. Approximately 135 women were enrolled in each stratum by physiologic periods (mid-pregnancy, late pregnancy, or lactation) and regions (south, central, or north). Plasma folate concentrations were measured by microbiological assay. The adjusted medians (interquartile ranges [IQR]) of folate concentration decreased from 28.8 (19.9 – 38.2) nmol/L in mid-pregnancy to 18.6 (13.2 – 26.4) nmol/L in late pregnancy, and to 17.0 (12.3 – 22.5) nmol/L in lactation (P for trend <0.001). Overall, lower folate concentrations were more likely to be observed in women residing in northern region, with younger age, higher pre-pregnancy BMI, lower education or multiparity, and in lactating women underwent a caesarean delivery or breastfed exclusively. In total, 380 (31.4%) women had a suboptimal folate status (folate concentration <13.5 nmol/L). Women of being at late pregnancy and lactation, residing in northern region, having multiparity and low education level had a higher risk of suboptimal folate status, while those with older age had a lower risk. In conclusion, maternal plasma folate concentrations decreased as pregnancy progressed, and influenced by geographic region and maternal social-demographic characteristics. Future studies are warranted to assess the necessity of folic acid supplementation during later pregnancy and lactation especially for women at a higher risk of folate depletion.