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While vitamin D 3  is recognized as a neuroactive steroid affecting both brain development and function, efficient analytical method in determining vitamin D 3  metabolites in the brain tissue is still lacking, and the relationship of vitamin D 3  status between serum and brain remains elusive. Therefore, we developed a novel analysis method by using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) to simultaneously quantify the concentrations of 25-hydroxyvitamin D 3  (25(OH)D 3 ) and 24,25-dihydroxyvitamin D 3  (24,25(OH) 2 D 3 ) in the serum and brain of rats fed with different dose of vitamin D 3 . We further investigated whether variations of serum vitamin D 3  metabolites could affect vitamin D 3  metabolite levels in the brain. Serum and brain tissue were analyzed by HPLC-MS/MS with electrospray ionization following derivatization with 4-phenyl-1,2,4-triazoline-3,5-dione (PTAD). The method is highly sensitive, specific, and accurate to quantify 25(OH)D 3  and 24,25(OH) 2 D 3  in animal brain tissue. Vitamin D 3  metabolites in brain tissue were significantly lower in rats fed with a vitamin D deficiency diet than in rats fed with high vitamin D 3  diet. There was also a strong correlation of vitamin D 3  metabolites in serum and brain. These results indicate that vitamin D 3  status in serum affects bioavailability of vitamin D 3  metabolites in the brain.

international journal of endocrinology

Simultaneous Quantification of 25-Hydroxyvitamin D<sub>3</sub>and 24,25-Dihydroxyvitamin D<sub>3</sub>in Rats Shows Strong Correlations between Serum and Brain Tissue Levels

Simultaneous Quantification of 25-Hydroxyvitamin D3and 24,25-Dihydroxyvitamin D3in Rats Shows Strong Correlations between Serum and Brain Tissue Levels

Simultaneous Quantification of 25-Hydroxyvitamin D₃and 24,25-Dihydroxyvitamin D₃in Rats Shows Strong Correlations between Serum and Brain Tissue Levels