Vitamin D is a fat-soluble vitamin that plays an important role in bone metabolism and seems to have some anti-inflammatory and immune-modulating properties. Cholesterol is the precursor of very important biochemical pathways including bile salts and other steroidogenic molecules like adrenal gland hormones, reproductive hormones, and vitamin D. In the present study, cholesterol and vitamin D test results from 13,150 patients admitted to our laboratory were analyzed over a one-year period retrospectively from the Laboratory Information System. In our study, we aimed to investigate the association between low cholesterol levels and vitamin D levels. Analyzed data revealed that low cholesterol levels (<200 mg/dl) were detected in 6,355 patients. In these patients, mean cholesterol level was 164.3 mg/dl while median vitamin D value was 20.4 ng/ml. There was a significant correlation (r=0.064, p<0.001) between low cholesterol levels and low vitamin D levels in the study group. The study group was divided into three subgroups according to age (<18, 18-35 and >35) and a significant correlation was detected between low cholesterol levels and low vitamin D (r=0.06, p<0.001) in patients >35 years of age. In this retrospective study, we determined a weak but significant correlation between vitamin D and cholesterol levels in particular >35 years of age. We think that this study data can be considered as a preliminary report for comprehensive studies that will determine the relationship between vitamin D and cholesterol levels in the general population.