Analysis of Vitamin D Components in Serum of Minors by UHPLC-MS/MS

Serum 25-hydroxyvitamin D [25(OH)D] concentration represents the body's reserves of vitamin D, which is mostly used by clinicians to evaluate the storage status of vitamin D in the body. The present study aimed to investigate the serum vitamin D components in different health status of minors to correctly evaluate the vitamin D storage in vivo. A total of 2,270 minors were included in the study, which was divided into healthy group (1,204 cases) and disease group (1,066 cases, including 270 short stature, 433 respiratory infections, 175 malnutrition and 188 tic disorder subjects). The levels of 25-hydroxyvitamin D₂ [25(OH)D₂] and 25-hydroxyvitamin D₃ [25(OH)D₃] were measured by UHPLC-MS/MS in all subjects, and the 25(OH)D₃ activity equivalents [25(OH)D₃-AE] and 25(OH)D were calculated. In addition, the 3-epi-25-hydroxyvitamin D₃ [3-epi-25(OH)D₃] concentrations of 278 subjects (including 147 healthy and 131 disease subjects) were measured by random sampling. 25(OH)D₂, 25(OH)D₃, 25(OH)D and 25(OH)D₃-AE levels in disease group were significantly lower than those in healthy group (p<0.001). According to the level of 25(OH)D, the sufficiency of vitamin D [25(OH)D≥30 ng/mL] was 65.4% in healthy group and 50.5% in disease group. When the 25(OH)D₂ activity was converted into 25(OH)D₃-AE, 53.2% of the patients in the healthy group had sufficiency vitamin D, and 39.1% in the disease group. The 3-epi-25(OH)D₃ level in the disease group was significantly lower than that in the healthy group (p<0.001). Not only the 25(OH)D, but also the both of 25(OH)D₂ and 25(OH)D₃ levels may overestimate the vitamin D status in subjects. For accurate evaluation, at least the serum levels of 25(OH)D₂, 25(OH)D₃ and 3-epi-25(OH)D₃ should be determined simultaneously.