Consistency evaluation of human serum 25-hydroxyvitamin D using liquid chromatography-tandem mass spectrometry and enzyme-linked immunosorbent assay

Abstract

Comparison of liquid chromatography-tandem mass spectrometry(LC-MS/MS) and enzyme-linked immunosorbent assay(ELISA) for the detection of 25(OH)D concentration in human serum and the diagnosis of vitamin D deficiency. In the serum pool of "National Institute for Nutrition and Health, Chinese Center for Disease Control and Prevention", 500 serum samples of women aged 18 to 45 years old were randomly selected, and 25(OH)D levels were measured by ELISA and LC-MS/MS for the same serum sample, respectively. The LC-MS/MS column was Waters XBridge BEH C_(18)(2. 1 mm×50 mm, 2. 5 μm). The correlation between the two method was tested by correlation analysis, regression analysis and consistency test. The Endocrine Society and Institute of Medicine recommendations were used to determine the deficiency of vitamin D, and the McNemar test, Kappa coefficient and diagnostic test were used to diagnose the consistency of vitamin D deficiency. The regression equation for the 25(OH)D concentration measured by the two method was y_(LC-MS/MS)=-0. 035+1. 007×x_(ELISA)(r=0. 877), and the average deviation between the two was 4. 48% and the intraclass correlation coefficient was 0. 87. The 25(OH)D concentration was less than 12 and 20 ng/mL as a criterion for vitamin D deficiency, and the Kappa coefficients were greater than 0. 60(0. 64 and 0. 74). When serum 25(OH)D level was detected by LC-MS/MS and ELISA, the correspondence of the two method was fine. Taking the "gold standard" LC-MS/MS method as a reference, the ELISA method was used to determine human vitamin D deficiency with good sensitivity and specificity, which can be used for the accurate and rapid detection of large-scale population samples.