Objective: To assess the prevalence of vitamin D deficiency and factors associated with serum vitamin D levels in adult women with class II/III obesity. Methods: We analysed baseline data from 128 adult women with class II/III obesity i.e. BMI ≥35 kg/m2 who participated in the DieTBra clinical trial. Sociodemographic, lifestyle, sun exposure, sunscreen, dietary intake of calcium and vitamin D, menopause, diseases, medication, and body composition data were analysed using multiple linear regression. Results: 128 women had mean BMI 45.53 ± 6.36, mean age 39.7 ± 8.75 kg/m2 and serum vitamin D 30.02 ng/ml ± 9.80. Vitamin D deficiency was 14.01%. There was no association between serum vitamin D levels and BMI, body fat percentage, total body fat and waist circumference. Age group (p = 0.004), sun exposure/day (p = 0.072), use of sunscreen (p = 0.168), inadequate calcium intake (p = 0.030), BMI (p = 0.192), menopause (p = 0.029) and lipid-lowering drugs (p = 0.150) were included in the multiple linear regression. The following remained associated with low serum vitamin D: being 40–49 years (p = 0.003); ≥50 years of age (p = 0.020) and inadequate calcium intake (p = 0.027). Conclusion: The prevalence of vitamin D deficiency was lower than expected. Lifestyle, sun exposure and body composition were not associated. Age over 40 years and inadequate calcium intake were significantly associated with low serum vitamin D levels.