Document Detail

HPLC method for 25-hydroxyvitamin D measurement: comparison with contemporary assays.
MedLine Citation:
PMID:  16574756     Owner:  NLM     Status:  MEDLINE    
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.
Gary L Lensmeyer; Donald A Wiebe; Neil Binkley; Marc K Drezner
Related Documents :
9528106 - Determination of vitamin b6 (pyridoxamine, pyridoxal and pyridoxine) in pork meat and p...
8380156 - In vivo metabolism of the vitamin d analog, dihydrotachysterol. evidence for formation ...
16526436 - Solid-phase extraction of vitamins a and e from animal feeds: a substitute for liquid--...
6292246 - Determination of vitamin d3 in cod-liver oil by high-performance liquid chromatography.
11489396 - Spectrophotometric determination of sparfloxacin in pharmaceutical formulations using b...
23773586 - Novel chiral core-shell silica microspheres with trans-(1r,2r)-diaminocyclohexane bridg...
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    
Clinical Toxicology Laboratory, University of Wisconsin Hospital and Clinics, Department of Pathology, University of Wisconsin-Madison, Madison, WI 53792, USA.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
25-Hydroxyvitamin D 2 / blood*
Calcifediol / blood*
Chromatography, High Pressure Liquid
Mass Spectrometry
Reg. No./Substance:
19356-17-3/Calcifediol; 21343-40-8/25-Hydroxyvitamin D 2
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

Previous Document:  Gap junctions and connexon hemichannels in human embryonic stem cells.
Next Document:  Peptide nucleic acid-based in situ hybridization assay for detection of parvovirus B19 nucleic acids...