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Americans are not reaching sufficient levels of serum 25-Hydroxy Vitamin D3 (25(OH)D3), and some groups are at a higher risk of developing vitamin D deficiency. Having a sufficient level of 25(OH)D3 can help prevent diseases like osteoporosis, certain types of cancers, cardiovascular disease, and hypertension. The data of 7479 American participants 18 years of age and older were derived from the NHANES 2001-2002 and NHANES 2003-2004 surveys. These participants were then analyzed to assess serum 25(OH)D3 levels. The variables used were age, gender, ethnicity, Body Mass Index (BMI), and smoking status. The statistical software, SUDDAN was used to control the weights of the samples, and least square means were calculated to control the confounding factors. The mean serum 25(OH)D3 levels (ng/ml) for Mexican males, Black males, White males and Mexican females, Black females and White females were 22.0 +/-0.6, 15.7 +/-0.5, 26.4+/- 0.5 and 19.0+/-0.4, 14.7+/-0.3, 26.5+/-0.4. The percent of Mexican males, Black males, White males and Mexican females, Black females and White females falling into the deficient category were 37.6%, 72.7%, 19.5% and 55.4%, 80.6%, 23.7%. Overall, the percent of males falling into the deficient category was 27.2% and for the females 33.9%. The mean serum 25(OH)D3 level for individuals greater than 70 years of age was the lowest overall, with the males' mean serum 25(OH)D3 level at 23.6+/-0.4 and the females' mean serum 25(OH)D3 level at 22.7+/-0.5. The mean serum 25(OH)D3 level for the obese BMI group was the lowest overall, with the males' mean serum 25(OH)D3 level at 23.2+/-0.5 and the females' mean serum 25(OH)D3 level at 21.0+/-0.5. The mean serum 25(OH)D3 level for the smokers was the lowest overall, with the male mean serum 25(OH)D3 level at 24.6+/-0.5 and the female mean serum 25(OH)D3 level at 23.2+/-0.6. In conclusion, the mean serum 25(OH)D3 level of Americans fell below the sufficient cut-off level of greater than 30 ng/ml, with only a small percentage of the population reaching this level.