Development and validation of a liquid chromatography-tandem mass spectrometry assay for serum 25-hydroxyvitamin D2/D3 using a turbulent flow online extraction technology

Abstract

Background: Vitamin D is important to health and disease. Liquid chromatography-tandem mass spectrometry (LC-MSMS) is considered the most accurate technology for quantification of serum 25-hydroxyvitamin D (25OHD) which is the best biomarker for estimating vitamin D nutritional status. Methods: Serum was mixed with acetonitrile containing hexadeuterated 25-hydroxyvitamin D 3 (d 6 -25OHD3) and centrifuged 10 min at 15,634=g. The supernatant was injected onto a turbulent flow preparatory column then transferred to a polar endcapped C18 analytical column. The mass spectrometer was set for positive atmospheric pressure chemical ionization. Results:The analytical cycle time was 5.5 min. Interand intra-assay CV for both analytes across three concentrations ranged from 3.8% to 14.2%. The method was linear from 3.0 to 283.6 nmol/L for 25-hydroxyvitamin D 3 (25OHD3) and 4.6 to 277.9 nmol/L for 25-hydroxyvitamin D 2 (25OHD2), with an accuracy of 88.7%-118.7% and 90.7%-100.3%, respectively. No carryover or ion suppression was observed. Comparison with a radioimmunoassay using patient specimens (ns527) showed a mean difference of 5.

Maternal Serum Vitamin D at 11–13 Weeks in Pregnancies Delivering Small for Gestational Age Neonates

“…This is particularly important for those variables that are known to be related to the risk for delivery of SGA neonates, including maternal racial origin, age, weight, height and smoking status [12]. Additional strengths of our study are firstly, examination of a large number of pregnancies resulting in delivery of SGA neonates within a narrow window at 11–13 weeks, which is emerging as the gestation of choice for risk assessment for a wide range of pregnancy complications [15], secondly, distinction between Caucasian women and those of African racial origin, and thirdly, measurement of vitamin D by LC-MSMS which is currently the most accurate technique for such estimation [16,17]. The main weakness resides in the design of the study which was case-control rather than prospective.…”

Normal Range of Maternal Serum Vitamin D at 11–13 Weeks’ Gestation

“…Recently, liquid chromatography tandem mass spectrometry technique (LC-MSMS) has been utilized and this is currently considered to be the most accurate technique for these measurements [19,20] . Other strengths of our study are the large number of cases examined and adjustment of the measured concentrations for the maternal factors that affect this measurement.…”

Clinical steroid mass spectrometry: A 45-year history culminating in HPLC–MS/MS becoming an essential tool for patient diagnosis

“…LC/MS/MS has now become the preferred technique for quantitation of the seco-sterol 25-hydroxy-vitamin D3 [64,71]. A candidate reference method (RMP) for progesterone has recently being published by Tai and colleagues at the NIST [72].…”