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Improved analysis of vitamin D metabolites in plasma using liquid chromatography tandem mass spectrometry, and its application to cardiovascular research
Author(s) -
Sandhu Jatinderpal K.,
Auluck Janica,
Ng Leong L.,
Jones Donald J. L.
Publication year - 2014
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.3208
Subject(s) - chromatography , chemistry , liquid chromatography–mass spectrometry , tandem mass spectrometry , mass spectrometry , sample preparation , extraction (chemistry) , selected reaction monitoring , vitamin , electrospray ionization , high performance liquid chromatography , coefficient of variation , electrospray , biochemistry
The accurate and specific measurement of vitamin D is increasingly important for determining the role of vitamin D in the pathogenesis of disease. Liquid chromatography coupled to tandem mass spectrometry (LC/MS/MS) has increasingly become the analytical modality of choice for the analysis of vitamin D. There are many advantages to using LC/MS/MS, such as high specificity and sensitivity to help distinguish the isomers of vitamin D. This rapid method, modified from a Waters Corporation application note, consists of minimal sample manipulation using liquid–liquid extraction and incorporates an internal standard. The supernatant is dried down and injected onto an ultra‐high‐performance liquid chromatography/electrospray ionization tandem mass spectrometer. The total analysis time is 10 min per injection, enabling high throughput of samples. This method also incorporates two commercial quality control standards to provide a robust system with acceptable coefficient of variation. The analysis of control and heart failure plasma samples showed significant differences in the levels of vitamin D 3 between these two groups; however, in the control group, there were individuals who were vitamin D deficient. Overall, the vitamin D 3 levels were higher in control samples than in heart failure individuals. As expected, vitamin D 2 levels were not observed in many of the samples analysed. This modified method is quick and incorporates an internal standard to allow for any loss in the extraction procedure. The method also includes quality control samples to enable assay standardization. The assay involves inexpensive pre‐sample clean‐up, aiding high throughput, which is important in many laboratories. Copyright © 2014 John Wiley & Sons, Ltd.