Aim: In this study, we aimed to investigate the association between serum vitamin D levels and dry eye signs and symptoms in postmenopausal women. Material and Method: Postmenopausal women with severe vitamin D deficiency (Group 1, n=30), moderate deficiency (Group 2, n=27) and those with optimal serum vitamin D levels (Group 3, n=30) were recruited for this cross-sectional study. Dry eye complaints were evaluated by the Ocular Surface Disease Index (OSDI) questionnaire and dry eye signs were evaluated by the Schirmer's test I, the Tear break up time (TBUT) test, and ocular surface fluorescein staining. Results: The mean vitamin D level of the Group 1 was 7.82± 1.80 ng/ml, that of the Group 2 was 15.88±1.70 ng/ml and Group 3 was 33.47±7.00 ng/ml The percentages of patients diagnosed as dry eye were 53.33% for Group 1, 44.44% for Group 2, and 33.33% for Group 3 (p₌0.29). There was no statistically significant difference between Group 1 (37.55 ± 19.20Group 2 (43.58 ±16.79), and Group 3 (40.87 ± 21.32) in respect to the mean OSDI scoresIn addition, there was no any statistically significant difference between the three groups in terms of the mean TBUT (Group 1: 7.73±3.25 secs, Group 2: 6.78±2.99 secs and Group 3: 6.70 ± 3.37 secs) and Schirmer's test I scores in (Group 1: 9.30 ± 4.86 mm, Group 2: 8.07 ± 4.09mm and Group 3: 9.40 ± 5.49mm) (p₌0.33 vs p₌0.72). Eight patients (26.7%) in Group 1, 5 patients (18.5%) in 19.20 Group 2, and 3 patients (10%) in Group 3 have positive corneal fluorescein staining (p₌0.25). Discussion: Serum vitamin D levels have no association with dry eye complaints evaluated by OSDI questionnaire and tear film deficiency and functional integrity detected by the tests in postmenopausal women.