Purpose To investigate the effect of vitamin D deficiency on tear break-up time (TBUT) and Schirmer test scores and to assess their relationship in non-Sjögren dry-eye patients. Methods Thirty-four patients with serum vitamin D deficiency and 21 control subjects with normal vitamin D levels were included in this study. The TBUT and Schirmer-1 test without topical anesthesia were performed to all patients. Results The mean TBUT were 5.18 ± 2.15 and 7.36 ± 3.10 s and Schirmer scores were 12.18 ± 6.44 and 18.57 ± 8.99 mm in the study and control groups, respectively. TBUT scores and Schirmer-1 results of the study group were significantly lower than the control group (P = 0.01 and 0.007, respectively). The mean vitamin D levels were 11.50 ± 1.8 ng/ml in the study group and 32.8 ± 8.72 ng/ml in control group (P = 0.001). Dry-eye symptoms were detected in all patients in the study group and 15% of the patients in the control group. Conclusions We demonstrated that vitamin D deficiency decreases the TBUT and Schirmer test values and may be associated with dry-eye symptoms in non-Sjögren syndrome.