Online citations, reference lists, and bibliographies.

Vitamin D Assays: Past And Present Debates, Difficulties, And Developments

W. D. Fraser, Anna M. Milan
Published 2012 · Medicine, Chemistry
Cite This
Download PDF
Analyze on Scholarcy
Clinical interest in Vitamin D and its purported roles not only in calcium and bone metabolism but in several other medical conditions (diabetes, cardiovascular disease, multiple sclerosis, cancer, psychiatric disorders, neuro-muscular disease) has led to a surge in laboratory requests for 25 hydroxy vitamin D and 1,25 dihydroxy vitamin D measurement. Circulating 25 hydroxy vitamin D concentration is routinely used as the best indicator of vitamin D status, but measurement of other metabolites, especially the physiologically active 1,25 dihyroxy vitamin D, are of clinical value. Over the last 40 years the development of assays for vitamin D and its metabolites from early competitive binding assays through to immunoassay and liquid chromatography aligned to mass spectrometry have demonstrated various analytical challenges, the advantages and disadvantages of each method are constantly changing with new technological developments. Immunoassay remains the predominant mode of measurement for 25-hydroxy vitamin D although problems with equimolar recovery of the D2 and D3 metabolites remain an issue. Standardisation of all assays has been improved but not resolved with the currently available reference materials as evidenced by the international vitamin D external quality assurance scheme, DEQAS. The choice of method for each laboratory remains a balance mainly between turn around time, convenience, cost and the specificity and accuracy of the information obtained. With increasing discussion and clinical interest surrounding other vitamin D metabolites the vitamin D assay debate is set to continue.
This paper references
Analytical evaluation of the new Abbott Architect 25-OH vitamin D assay.
Etienne Cavalier (2012)
Determination of 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 in human plasma using high-pressure liquid chromatography.
John A. Eisman (1977)
Development and certification of a standard reference material for vitamin D metabolites in human serum.
Karen W Phinney (2012)
Measurement of 25-hydroxyvitamin D3 in serum.
Roger Bouillon (1976)
Analytical and clinical validation of a radioimmunoassay for the measurement of 1,25 dihydroxy vitamin D.
Diana R Clive (2002)
Plasma concentrations of vitamin D3 and its metabolites in the rat as influenced by vitamin D3 or 25-hydroxyvitamin D3 intakes.
Rj Shephard (1980)
Sensitive measurement of serum 1α,25-dihydroxyvitamin D by liquid chromatography/tandem mass spectrometry after removing interference with immunoaffinity extraction.
Chao Annie Yuan (2011)
Increasing liquid chromatography-tandem mass spectrometry throughput by mass tagging: a sample-multiplexed high-throughput assay for 25-hydroxyvitamin D2 and D3.
Brian C. Netzel (2011)
Measurement of 25-hydroxyvitamin D3 (25OHD3) and 25-hydroxyvitamin D2 (25OHD2) in human serum using liquid chromatography-tandem mass spectrometry and its comparison to a radioimmunoassay method.
Huiping Chen (2008)
Current status of clinical 25-hydroxyvitamin D measurement: an assessment of between-laboratory agreement.
N Binkley (2010)
A modified radioimmunoassay for 1,25-dihydroxycholecalciferol.
T. Kenney Gray (1981)
Underestimation of serum 25-hydroxyvitamin D by the Nichols Advantage Assay in patients receiving vitamin D replacement therapy.
Pamela Leventis (2005)
Development and optimization of simplified LC-MS/MS quantification of 25-hydroxyvitamin D using protein precipitation combined with on-line solid phase extraction (SPE).
Denis Thibeault (2012)
Determination of 25-hydroxyvitamin D in human plasma using high-performance liquid chromatography--tandem mass spectrometry.
Naoko Tsugawa (2005)
Determination of vitamin D status by radioimmunoassay with an 125I-labeled tracer.
B W Hollis (1993)
Effect of a 300 000-IU loading dose of ergocalciferol (Vitamin D2) on circulating 1,25(OH)2-vitamin D and fibroblast growth factor-23 (FGF-23) in vitamin D insufficiency.
Christopher D. Turner (2013)
Isotope dilution ultra performance liquid chromatography-tandem mass spectrometry method for simultaneous measurement of 25-hydroxyvitamin D2, 25-hydroxyvitamin D3 and 3-epi-25-hydroxyvitamin D3 in human serum.
Rosemary L. Schleicher (2011)
Comparison of commercially available (125)I-based RIA methods for the determination of circulating 25-hydroxyvitamin D.
B W Hollis (2000)
Evaluation of 25-hydroxy vitamin D assay on the immunodiagnostic systems iSYS analyser
Zee N Cluse (2012)
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.
Susan S-C Tai (2010)
Characterization of vitamin D3 metabolites using continuous-flow fast atom bombardment tandem mass spectrometry and high-performance liquid chromatography.
Bernice Yeung (1993)
Development of a Method for the Quantification of 1α,25(OH)2–Vitamin D3 in Serum by Liquid Chromatography Tandem Mass Spectrometry without Derivatization
Bruno Casetta (2010)
A radioimmunoassay for 1,25-dihydroxycholecalciferol.
Roger Bouillon (1980)
Assay of 1,25-dihydroxy vitamin D3 by isotope dilution--mass fragmentography.
Ingemar Björkhem (1979)
A competitive protein-binding assay for 25-hydroxycholecalciferol and 25-hydroxyergocalciferol in serum.
Michael A. Preece (1974)
Radioreceptor assay for 1 alpha,25-dihydroxyvitamin D3.
P. F. Brumbaugh (1974)
1,25-Dihydroxyvitamin D3-26,23-lactone interferes in determination of 1,25-dihydroxyvitamin D by RIA after immunoextraction.
B W Hollis (1995)
C-3 epimers can account for a significant proportion of total circulating 25-hydroxyvitamin D in infants, complicating accurate measurement and interpretation of vitamin D status.
Ravinder Singh (2006)
Improved radioimmunoassay for vitamin D and its use in assessing vitamin D status.
B. W. Hollis (1985)
Pharmacokinetics of vitamin D toxicity.
Glenville Jones (2008)
Sensitive analysis of 1alpha,25-dihydroxyvitamin D3 in biological fluids by liquid chromatography-tandem mass spectrometry.
Anne-Marie Kissmeyer (2001)
Candidate reference measurement procedures for serum 25-hydroxyvitamin D3 and 25-hydroxyvitamin D2 by using isotope-dilution liquid chromatography-tandem mass spectrometry.
Hedwig C M Stepman (2011)
Ultrasensitive quantification of serum vitamin D metabolites using selective solid-phase extraction coupled to microflow liquid chromatography and isotope-dilution mass spectrometry.
Xiaotao Duan (2010)
Structure-function relationships in the vitamin D endocrine system.
Roger Bouillon (1995)
Clinical applications for vitamin D assays: what is known and what is wished for.
Michael Kleerekoper (2011)
Circulating 25-hydroxyvitamin D in man.
John G. Haddad (1974)
Extrarenal Expression of 25-Hydroxyvitamin D 3-1 a-Hydroxylase *
Daniel Zehnder (2001)
Measurement of circulating vitamin D in man.
Thomas L. Clemens (1982)
Intestinal cholecalciferol absorption in the elderly and in younger adults.
J. M. Barragry (1978)
Accuracy of 6 Routine 25-Hydroxyvitamin D Assays: Influence of Vitamin D Binding Protein Concentration
Michael G. Bissell (2013)
Competitive protein-binding radioassay for 25-hydroxycholecalciferol.
John G. Haddad (1971)
Radioimmunoassay of 1,25-dihydroxy vitamin D2: studies on the metabolism of vitamin D2 in man.
Laurence J. Fraher (1983)
High-performance liquid chromatographic assay for 25-hydroxyvitamin D3 in serum.
Terry J. Gilbertson (1977)
Misleading measures in Vitamin D analysis: A novel LC-MS/MS assay to account for epimers and isobars
Iltaf Shah (2011)
Serum vitamin D2 and vitamin D3 metabolite concentrations and absorption of vitamin D2 in elderly subjects.
Thomas L. Clemens (1986)
How accurate are assays for 25-hydroxyvitamin D? Data from the international vitamin D external quality assessment scheme.
Graham D. Carter (2004)
Issues of methodology, standardization and metabolite recognition for 25-hydroxyvitamin D when comparing the DiaSorin radioimmunoassay and the Nichols Advantage automated chemiluminescence protein-binding assay in hip fracture cases
Paul Glendenning (2003)
Human anti-animal interference in DiaSorin Liaison total 25(OH)-vitamin D assay: towards the end of a strange story?
Etienne Cavalier (2012)
Candidate reference method for the quantification of circulating 25-hydroxyvitamin D3 by liquid chromatography-tandem mass spectrometry.
Michael Vogeser (2004)
Proficiency testing of 25-Hydroxyvitamin D (25-OHD) assays
Ghangis D. Carter (2010)
Ion suppression in the determination of clenbuterol in urine by solid-phase extraction atmospheric pressure chemical ionisation ion-trap mass spectrometry.
Mischa W J van Hout (2003)
Uskokovic MR (1980) A radioimmunoassay for 1,25-dihydroxycholecalciferol
R Bouillon (1980)
State-oftheArt oftheArt oftheArt theArt theArt Vitamin D Assays : A Comparison of Automated Immunoassays with Liquid Chromatography – Tandem Mass Spectrometry Methods
Christopher-John L Farrell (2012)
Metabolic profiling of major vitamin D metabolites using Diels–Alder derivatization and ultra-performance liquid chromatography–tandem mass spectrometry
Pavel A. Aronov (2008)
Metabolism and excretion of 3H-1,25-(OH)2-vitamin D3 in healthy adults.
Richard W. Gray (1978)
Current assays overestimate 25-hydroxyvitamin D3 and underestimate 25-hydroxyvitamin D2 compared with HPLC: need for assay-specific decision limits and metabolite-specific assays
Paul Glendenning (2006)
On-line post-column Diels-Alder derivatization for the determination of vitamin D3 and its metabolites by liquid chromatography/thermospray mass spectrometry.
Rob J. Vreeken (1993)
Specific estimation of 24,25-dihydroxyvitamin D in plasma by gas chromatography-mass spectrometry.
Ruth D. Coldwell (1984)
Improved methodology for the analysis of plasma vitamin D metabolites.
Ronald L. Horst (1990)
Analytical and clinical validation of the 25 OH vitamin D assay for the LIAISON automated analyzer.
Diana L. Ersfeld (2004)
25-hydroxyvitamin D: a difficult analyte.
Graham D. Carter (2012)
25-hydroxyvitamin D measurement, 2009: a review for clinicians.
N Binkley (2009)
The effect of renal function on changes in circulating concentrations of 1,25-dihydroxycholecalciferol after an oral dose.
Socrates E Papapoulos (1982)
Standardization of vitamin D assays: art or science?
W. D. Fraser (2009)
Development and validation of a liquid chromatography-tandem mass spectrometry assay for serum 25-hydroxyvitamin D2/D3 using a turbulent flow online extraction technology
Dustin R. Bunch (2009)
Vitamin D metabolites and calcium absorption in severe vitamin D deficiency.
Allan G. Need (2008)
Competitive protein-binding assay for 25-hydroxycholecalciferol.
Samuel Edelstein (1974)
Importance of the isolation of 25-hydroxyvitamin D before assay.
L M Dorantes (1978)
Acute effects of intravenous 1 alpha-hydroxycholecalciferol on parathyroid hormone, osteocalcin and calcitriol in man.
Stephen J. Gallacher (1994)
Evaluation of a 1,25-dihydroxyvitamin D enzyme immunoassay.
Isolde Seiden-Long (2007)
NHANES monitoring of serum 25-hydroxyvitamin D: a roundtable summary.
Elizabeth A. Yetley (2010)
Measurement of plasma 25-hydroxycholecalciferol in man.
Françis Bayard (1972)
Quantification of circulating 1,25-dihydroxyvitamin D by radioimmunoassay with 125I-labeled tracer.
B W Hollis (1996)
Assay of vitamins D2 and D3, and 25-hydroxyvitamins D2 and D3 in human plasma by high-performance liquid chromatography.
Grace Jones (1978)
Routine isotope-dilution liquid chromatography-tandem mass spectrometry assay for simultaneous measurement of the 25-hydroxy metabolites of vitamins D2 and D3.
Zoë J. Maunsell (2005)
A simple automated solid-phase extraction procedure for measurement of 25-hydroxyvitamin D3 and D2 by liquid chromatography-tandem mass spectrometry
Susan Knox (2009)
Low circulating vitamin D in obesity
Yair Liel (2007)
Use of a common standard improves the performance of liquid chromatography-tandem mass spectrometry methods for serum 25-hydroxyvitamin-D
Graham D. Carter (2009)
Competitive binding assay for vitamin D and 25-OH vitamin D.
Richard Belsey (1971)
Measurement of Plasma 1,25 Dihydroxyvitamin D Using a Novel Immunoextraction Technique and Immunoassay with Iodine Labelled Vitamin D Tracer
W. D. Fraser (1997)
A microassay for 1,25-dihydroxyvitamin D not requiring high performance liquid chromatography: application to clinical studies.
Timothy A Reinhardt (1984)
Serum 25-hydroxyvitamin D measurement in a large population survey with statistical harmonization of assay variation to an international standard.
Elina Hyppönen (2007)
HPLC method for 25-hydroxyvitamin D measurement: comparison with contemporary assays.
Gary L. Lensmeyer (2006)
The C-3 epimer of 25-hydroxyvitamin D(3) is present in adult serum.
Gary L. Lensmeyer (2012)

This paper is referenced by
Measurements of 25-hydroxyvitamin D concentrations in archived dried blood spots are reliable and accurately reflect those in plasma.
Alicia K. Heath (2014)
Vitamin D supplementation has no effects on progression of motor dysfunction in amyotrophic lateral sclerosis (ALS)
Francesca Trojsi (2019)
Is there an optimal vitamin D status for immunity in athletes and military personnel?
Cheng-Shiun He (2016)
Effects of Iron on Vitamin D Metabolism: A Systematic Review
Fatemeh Azizi-soleiman (2016)
Vitamin D Testing in Clinical Settings: Methodologies, Accuracy and Standardization
Fakhri B Saida (2018)
Vitamin D Measurements — Facts and Fancies
Tar Choon Aw (2013)
Subclinical and asymptomatic parathyroid disease: implications of emerging data.
David P. Macfarlane (2013)
Vitamin D in Renal Transplantation – from Biological Mechanisms to Clinical Benefits
Reuben McGregor (2014)
Critical evaluation of assays for vitamin D status.
Fernando Granado Lorencio (2013)
Optimization of Chemical Syntheses of Vitamin D C3-Epimers.
Lars Kattner (2016)
Analysis of vitamin D status at two academic medical centers and a national reference laboratory: result patterns vary by age, gender, season, and patient location
Jonathan R Genzen (2013)
Method evaluation study of a new generation of vitamin D assays
Dietmar Enko (2015)
Impact of Vitamin D Supplementation on Arterial Vasomotion, Stiffness and Endothelial Biomarkers in Chronic Kidney Disease Patients
Nihil Chitalia (2014)
Evaluation of the 25-hydroxy vitamin D assay on a fully automated liquid chromatography mass spectrometry system, the Thermo Scientific Cascadion SM Clinical Analyzer with the Cascadion 25-hydroxy vitamin D assay in a routine clinical laboratory
Sally C Benton (2019)
Automated measurement of 25-OH Vitamin D on the LUMIPULSE® G1200: analytical verification and method comparison
Marina Parra (2016)
Screening for Vitamin D Deficiency: A Systematic Review for the U.S. Preventive Services Task Force
Erin S LeBlanc (2015)
Prevalence and Predictors of Low Serum 25-Hydroxyvitamin D among Female African-American Breast Cancer Survivors.
Patricia M. Sheean (2018)
Institutional Repository The e ect of 14 weeks of vitamin D 3 supplementation on antimicrobial peptides and proteins in athletes
Cheng-Shiun He (2017)
Association of preconception serum 25-hydroxyvitamin D concentrations with livebirth and pregnancy loss: a prospective cohort study.
Sunni L. Mumford (2018)
A review and clinical summary of vitamin D in regard to bone health and athletic performance
Sara Neal (2015)
Social Disadvantage Is Associated With Lower Vitamin D Levels in Older People and There Is No Surrogate for Its Measurement
Adrian H Heald (2017)
A High Throughput Universal Vitamin D Assay for Automated Chemistry Analyzers
Saida Fb (2019)
Vitamin D in the Prevention and Treatment of Osteoarthritis: From Clinical Interventions to Cellular Evidence
Clara Yongjoo Park (2019)
Magnesium Supplementation in Vitamin D Deficiency
Pramod Reddy (2019)
Milk fortified with vitamin D could reduce the prevalence of vitamin D deficiency among Japanese female college students
Yoshio Suzuki (2014)
Vitamin D in Fetal Development: Findings From a Birth Cohort Study
Prue H Hart (2015)
Older Swedish Adults with High Self-Perceived Health Show Optimal 25-Hydroxyvitamin D Levels Whereas Vitamin D Status Is Low in Patients with High Disease Burden
Martin Carlsson (2016)
Vitamin D and exercise performance : influence of simulated sunlight and oral supplementation
Alexander T. Carswell (2019)
Serum free and bio-available 25-hydroxyvitamin D correlate better with bone density than serum total 25-hydroxyvitamin D
Martin Solvang Johnsen (2014)
Artefactual 25-OH vitamin D concentration in multiple myeloma
Michal Ws Ong (2017)
Iron-based phosphate binders – a new element in management of hyperphosphatemia
Amy Barton Pai (2016)
Vitamin D Measurement, the Debates Continue, New Analytes Have Emerged, Developments Have Variable Outcomes
W. D. Fraser (2019)
See more
Semantic Scholar Logo Some data provided by SemanticScholar