Document Detail


HPLC method for 25-hydroxyvitamin D measurement: comparison with contemporary assays.
MedLine Citation:
PMID:  16574756     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: The concentration of 25-hydroxyvitamin D [25(OH)D] in serum has been designated the functional indicator of vitamin D (VitD) nutritional status. Unfortunately, variability among 25(OH)D assays limits clinician ability to monitor VitD status, supplementation, and toxicity. METHODS: We developed an HPLC method that selectively measures 25-hydroxyvitamin D2 [25(OH)D2] and D3 [25(OH)D3] and compared this assay with a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, a competitive protein-binding assay (CPBA) on the Nichols Advantage platform, and an RIA from Diasorin. RESULTS: For the new HPLC assay, between-run CVs were 2.6%-4.9% for 25(OH)D3 and 3.2%-13% for 25(OH)D2; recoveries were 95%-102%; and the assay was linear from 5 microg/L to at least 200 microg/L. Comparison data were as follows: for HPLC vs LC-MS/MS, y = 1.01x - 4.82 microg/L (Sy/x = 4.93 microg/L; r = 0.996) for 25(OH)D3, and y = 0.902x - 0.566 microg/L (Sy/x = 2.56 microg/L; r = 0.9965 for 25(OH)D2; for HPLC vs Diasorin RIA, y = 0.709x - 5.86 microg/L (Sy/x = 7.35 microg/L; r = 0.7509); and for HPLC vs Nichols Advantage CPBA, y = 1.00x - 3.60 microg/L (Sy/x = 32.7 microg/L; r = 0.6823). CONCLUSIONS: The new HPLC method is reliable, robust, and has advantages compared with the Nichols Advantage CPBA and the Diasorin RIA. The Nichols Advantage CPBA overestimated or underestimated 25(OH)D concentrations predicated on the prevailing metabolite present in patients' sera.
Authors:
Gary L Lensmeyer; Donald A Wiebe; Neil Binkley; Marc K Drezner
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Publication Detail:
Type:  Comparative Study; Journal Article     Date:  2006-03-30
Journal Detail:
Title:  Clinical chemistry     Volume:  52     ISSN:  0009-9147     ISO Abbreviation:  Clin. Chem.     Publication Date:  2006 Jun 
Date Detail:
Created Date:  2006-05-25     Completed Date:  2006-06-27     Revised Date:  2007-02-02    
Medline Journal Info:
Nlm Unique ID:  9421549     Medline TA:  Clin Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1120-6     Citation Subset:  IM    
Affiliation:
Clinical Toxicology Laboratory, University of Wisconsin Hospital and Clinics, Department of Pathology, University of Wisconsin-Madison, Madison, WI 53792, USA. g.lensmeyer@hosp.wisc.edu
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MeSH Terms
Descriptor/Qualifier:
25-Hydroxyvitamin D 2 / blood*
Calcifediol / blood*
Chromatography, High Pressure Liquid
Humans
Mass Spectrometry
Radioimmunoassay
Chemical
Reg. No./Substance:
19356-17-3/Calcifediol; 21343-40-8/25-Hydroxyvitamin D 2
Comments/Corrections
Comment In:
Clin Chem. 2006 Dec;52(12):2304-5; author reply 2305-6   [PMID:  17138852 ]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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