Evaluation of correlations between serum and urinary vitamin D metabolites using LC-MS/MS

Vitamin D (VD) is involved in bone health, immunity, cardiovascular function, and cancer prevention. Recent studies suggested the potential of 24,25-dihydroxyvitamin D3 [24,25(OH)2D3] and the VD metabolite ratio (VMR = [24,25(OH)2D3/25(OH)D] × 100) as indicators of VD levels. An assessment of VD levels relies on serum measurements. The present study investigated the utility of urinary VD metabolites as a non-invasive alternative and their potential as surrogate markers for serum VD metabolites. We used residual serum and urine from the annual health check-ups of employees at Shinshu University Hospital, Japan (n = 506). Urine samples were mixed with β-glucuronidase. Serum and urine were processed via solid-phase extraction, followed by DAP-PA derivatization. LC-MS/MS was used to quantify serum and urinary 25(OH)D3, 25(OH)D2, 3-epi-25(OH)D3, and 24,25(OH)2D3 and urinary 23,25(OH)2D3. Serum whole parathyroid hormone (PTH) levels were measured, and urinary VD metabolites were adjusted for urinary creatinine. We also investigated whether serum and urinary VD metabolites correlated with PTH. The results showed that urinary 24,25(OH)2D3 and 23,25(OH)2D3 strongly correlated with serum 25(OH)D [25(OH)D3 + 25(OH)D2] (all ρ > 0.7, p < 0.001). Urinary 24,25(OH)2D3, 23,25(OH)2D3, and VMR had stronger inverse correlations with whole PTH (ρ = -0.297, -0.274, -0.308, respectively; all p < 0.001), whereas serum 25(OH)D showed a weak correlation (ρ = -0.174, p < 0.001). In conclusion, urinary VD metabolites, particularly 24,25(OH)2D3, 23,25(OH)2D3 and the VMR, correlated with serum values and PTH, suggesting their potential as non-invasive markers for the VD status.