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Quantitation of 25-hydroxyvitamin D2 and D3 in serum and plasma by LCMS/MS
Dayong Lee1, Timothy J Garrett, Bruce A Goldberger
1UF Health Pathology Laboratories, Department of Pathology, Immunology & Laboratory Medicine, University of Florida College of Medicine, Gainesville, FL, USA.
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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