Quantitation of 25-hydroxyvitamin D2 and D3 in serum and plasma by LCMS/MS.

Bioanalysis

UF Health Pathology Laboratories, Department of Pathology, Immunology & Laboratory Medicine, University of Florida College of Medicine, Gainesville, FL, USA.

Published: September 2015

Objective: We developed and validated an analytical method for quantifying 25-hydroxyvitamin D2 (25OHD2) and 25-hydroxyvitamin D3 (25OHD3) in serum and plasma.

Methods: Samples, pretreated with zinc sulfate and methanol, were extracted with hexane. Separation was achieved via UHPLC and 25OHD quantification was accomplished by a triple quadrupole mass spectrometer.

Results: Imprecision was 3.6-15.1%CV and bias 88.0-126.0%. Extraction efficiency was 76.5-110.5%, whereas the matrix effect ranged from -46.7 to -32.0%. The method was applied to authentic specimens. The results showed no significant difference between serum and plasma; strong correlation with paired values from an external laboratory; and analyte stability for 15 days.

Conclusion: This method provides reliable and accurate measurement of 25OHD for use in clinical practice.

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Source
http://dx.doi.org/10.4155/bio.14.229DOI Listing

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