Analysis of 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 in Serum Using Ultra-performance Liquid Chromatography with Tandem Mass Spectrometry

Content Provider	Semantic Scholar
Author	Lee, Yong-Wha
Copyright Year	2013
Abstract	Background: Measurement of serum 25-hydroxyvitamin D metabolites (25OHD) may be useful in the laboratory diagnosis and treatment of vitamin D deficiency. A standardized liquid chromatography-tandem mass spectrometry (LC-TMS) assay has been used for accurate analysis of 25-hydroxyvitamin D levels. We evaluated the performance of a recently developed ultra-performance liquid chromatography combined with tandem mass spectrometry (UPLC-TMS) for the simultaneous quantification of 25OHD3 and 25OHD2 in human serum. Materials and Methods: We established the measurement method of 25-hydroxyvitamin D using UPLC-TMS and evaluated the precision and made a comparison between the UPLC-TMS and the chemiluminescence immunoassay (CLIA). Proportions of vitamin D deficiency in a local population and the spectrum of vitamin D2 measured were investigated. Results: UPLC-TMS revealed within-run precision and between-run precision of less than 4% and showed a bias of less than 5%. The CLIA measurements of 25OHD showed a correlation of CLIA = 0.86 × UPLC- TMS + 1.16 (r 2 = 0.98). The means±SD of serum 25OHD, 25OHD2, and 25OHD3 in 1,047 individuals were 12.8±9.3 ng/mL (0.5-62.7 ng/mL), 0.6±1.6 ng/mL (0-22.4 ng/mL), and 12.2±9.2 ng/mL (0.4-62.4 ng/mL), respectively. The proportions of vitamin D insufficiency and deficiency were 34.4% (360/1047) and 48.6% (509/1047), respectively. Discussions: The newly developed 25OHD assay using the MSMS Vitamin D kit based on the UPLC-TMS assay showed excellent performance and is suitable for routine clinical measurements of serum 25OHD2 and 25OHD3. We expect that this method will contribute to improved accuracy and reduced interlaboratory variation versus other methods for 25OHD measurement. (Yong-Wha Lee. Analysis of 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 in Serum Using Ultra- performance Liquid Chromatography with Tandem Mass Spectrometry. Life Sci J 2013; 10(3): 1648-1652). (ISSN: 1097-8135). http://www.lifesciencesite.com 247
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