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The aim is to develop a candidate reference measurement procedure for accurately determining 25-hydroxyvitamin D levels in human serum.

February 5, 2010Analytical ChemistryOpen Access

Development of a Candidate Reference Measurement Procedure for the Determination of 25-Hydroxyvitamin D₃ and 25-Hydroxyvitamin D₂ in Human Serum Using Isotope-Dilution Liquid Chromatography−Tandem Mass Spectrometry

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The aim is to develop a candidate reference measurement procedure for accurately determining 25-hydroxyvitamin D levels in human serum.
Developed using isotope-dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS).
Internal standards 25(OH)D(3)-d(3) and 25(OH)D(2)-d(3) were used for accuracy.
Simultaneous extraction from serum and chromatographic separation achieved using a cyano column.
Recovery rates for added 25(OH)D(3) and 25(OH)D(2) were between 99.0% and 101.0%.
Achieved precision indicated by coefficients of variation of 0.2-0.6% for levels >1 ng/g.
Limit of detection for 25(OH)D was 40 pg on column, demonstrating good accuracy and low interference.

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Randomized trial develops a reference method for measuring vitamin D levels in serum, suggesting improved accuracy in clinical testing.

Tai et al. (2010) studied this question.

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