The precursor of the active form of vitamin D, 25-hydroxyvitamin D (25(OH)D), is recognized as the optimal indicator of vitamin D status. Vitamin D3 undergoes conversion through a multitude of enzymatic reactions described within the paper, and vitamin D levels are dependent on many factors including the vitamin D binding protein (DBP). The free hormone hypothesis postulates that protein-bound hormones are not biologically available and that unbound hormones are biologically active. The majority of circulating 25(OH)D and 1,25-dihydroxyvitamin D is tightly bound to DBP and albumin, with less than 1% circulating in an unbound form. As a result, factors affecting DBP alter the interpretation of 25(OH)D levels. The aim of this review is to assess the current methodology used to measure total and free 25(OH)D, and DBP. Additionally, we analyze the effects of other endocrine hormones and disease processes on DBP levels and subsequently, the interpretation of 25(OH)D levels.

Abbreviations

CF = cystic fibrosis DBP = vitamin D binding protein ELISA = enzyme-linked immunosorbent assay ESLD = end-stage liver disease HC = hormone contraceptives iPTH = intact parathyroid hormone LC-MS = liquid chromatography-mass spectrometry MS = multiple sclerosis 25(OH)D = 25-hydroxyvitamin D PHPT = primary hyperparathyroidism RIA = radioimmunoassay.