The prevalence of vitamin D deficiency differs in various parts of the world based on ethnicity, latitude, environmental factors, and sociocultural practices; vitamin D deficiency is more common in pregnant women than non-pregnant women. Vitamin D deficiency is a common concern during pregnancy due to increased demands on the mother’s body to support fetal development. It is a significant public health problem for pregnant women in Libya. The present study aimed to assess the vitamin D status among pregnant women in the Surman region in Western Libya. An observational study was conducted among 200 subjects (100 pregnant women and 100 non-pregnant women) in the Surman region in Western Libya, over seven months from the 1st September 2024 to 31st March 2025. Serum 25-hydroxyvitamin D levels were measured using an enzyme immunoassay method. The associations of the levels of 25 hydroxyvitamin D with age groups and pregnancy periods were assessed using the Chi-square test, ANOVA test, and Person correlation by SPSS version 25. The results showed that the serum vitamin D levels were <10 ng/mL, (10-20 ng/mL), and (21-29 ng/mL) in 79%, 17%, 4% of pregnant women, respectively. The mean serum vitamin D concentrations were significantly (P<0.01) decreased in pregnant (8.611±0.47 ng/ml) compared to non-pregnant women (18.874±0.90 ng/ml). These decreases were significant (P<0.05) in the second period of pregnancy (7.795±0.54 ng/ml) compared to the first period of pregnancy (10.648±1.22 ng/ml), and significantly (P<0.01) decreased (7.649±0.50 ng/ml) in the third period of pregnancy compared to the first period of pregnancy (10.648±1.22 ng/ml). The mean serum vitamin D concentrations were significantly (P<0.01) decreased, 8.293±0.72, 9.22±0.82, and 8.614±0.47 ng/ml in age groups, (20-29), (30-39), and (>39) years in pregnant women compared with the same age groups in non-pregnant women, 16.05±1.88, 20.13±1.82, and 19.09±0.63 ng/ml, respectively. The mean serum vitamin D concentrations were significantly (P<0.01) decreased in severe deficiency vitamin D pregnant women (6.821±0.20 ng/ml) compared to insufficient vitamin D in the same group (15.770±1.34 ng/ml). The mean serum vitamin D concentration in insufficient vitamin D in pregnant women significantly decreased (15.770±1.34 ng/ml) compared with that in insufficient vitamin D in non-pregnant Women (19.372±0.69 ng/ml). A significant negative correlation (r = -0.586) at (P<0.01) was observed between vitamin D concentrations with Pregnancy and a significant negative correlation (r = -0.237) with the periods of pregnancy at (P<0.05). The results showed that vitamin D deficiency was extremely common among pregnant women in the Surman region of Western Libya. A significant negative correlation was observed between vitamin D concentrations with Pregnancy and the periods of pregnancy. Pregnant women should consume foods high in vitamin D and expose their skin to sunlight for a sufficient amount of time to boost their body's natural synthesis of vitamin D. The information might be used to create health promotion programs that maximize pregnant women's vitamin D levels. More study is needed to determine if untreated vitamin D deficiency in the early stages of pregnancy has an impact on the mother and the unborn child and to assist Libyan women in selecting suitable therapy. Our findings highlight the need for increased awareness and potential interventions to address vitamin D inadequacy in pregnant women to enhance outcomes for both mothers and babies................ Keywords:............... Vitamin D deficiency, Vitamin D Levels, Pregnant women, Correlation with pregnancy and its periods, Surman region, Western Libya.