Acylation derivatization based LC-MS analysis of 25-hydroxyvitamin D from finger-prick blood

Vitamin D metabolite analysis possessed significant clinical value for the pediatric department. However, invasive venipuncture sampling and high blood consumption inflicted much suffering on patients. For alleviation, we carried out a LC-MS method for 25-hydroxyvitamin D quantification in only 3 μl of plasma from the considerably less invasive finger-prick blood samples. To improve sensitivity, acylation on C3-hydroxyl (by isonicotinoyl chloride) rather than Diels-Alder adduction on s-cis-diene structure was for the very first time introduced into vitamin D metabolite derivatization. Compared with the existing derivatization approaches, this novel strategy not only prevented isomer interference, but also exhibited higher reacting throughput. For certification, the methodology was systematically validated and showed satisfying consistency with SRM927a. During clinical application, we found a convincing correlation between 25-hydroxyvitamin D and indirect/total bilirubin in jaundiced newborns. Such an observation indicated that vitamin D supplementation may help to achieve optimal outcomes in neonatal jaundice.