The Impact of Sample Type on Vitamin D Quantification and Clinical Classification during Pregnancy

Measurement of vitamin D status has significant use in clinical and research settings, including during pregnancy. We aimed to assess the agreement of total 25-hydroxyvitamin D (25(OH)D) concentration, and its three analytes (25-hydroxyvitamin D (25(OH)D ), 25-hydroxyvitamin D (25(OH)D ) and Epi-25-hydroxyvitamin D (Epi-25(OH)D )), in plasma and serum samples collected during pregnancy, and to examine the proportion of women who change vitamin D status category based on sample type. Matching samples were collected from = 114 non-fasting women between 12-25 weeks gestation in a clinical trial in Newcastle, Australia. Samples were analysed by liquid chromatography-tandem mass-spectrometry (LC-MS/MS) to quantify total 25(OH)D and its analytes and examined using Bland-Altman plots, Pearson correlation (r), intraclass correlation coefficient and Cohen's Kappa test. Serum total 25(OH)D ranged from 33.8-169.8 nmol/L and plasma ranged from 28.6-211.2 nmol/L. There was a significant difference for total 25(OH)D based on sample type (measurement bias 7.63 nmol/L for serum vs plasma (95% Confidence Interval (CI) 5.36, 9.90, ≤ 0.001). The mean difference between serum and plasma concentrations was statistically significant for 25(OH)D (7.38 nmol/L; 95% CI 5.28, 9.48, ≤ 0.001) and Epi-25(OH)D (0.39 nmol/L; 95% CI 0.14, 0.64, = 0.014). Of 114 participants, 28% were classified as vitamin D deficient (