Fast Separation of 25-Hydroxyvitamin D3 from 3-Epi-25-Hydroxyvitamin D3 in Human Serum by Liquid Chromatography–Tandem Mass Spectrometry: Variable Prevalence of 3-Epi-25-Hydroxyvitamin D3 in Infants, Children, and Adults

Ouweland, Johannes M.W. van den; Beijers, Antonius M.; Daal, Henny van

doi:10.1373/clinchem.2011.170282

Cited by 50 publications

(42 citation statements)

References 5 publications

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“…The recoveries were 83.8%-109.2% (mean, 95.9%). The lower limits of quantification for D 3 and D 2 were 3.0 ng/mL (7.5 nmol/L) and 1.0 ng/mL (2.5 nmol/ L), respectively. We tested the 2011 College of American Pathologists proficiency testing set ABVD-01-05 with the assay, and all results were within Ϯ25% of target values (mean bias, Ϫ1.2%).…”

Section: -Hydroxyvitamin D (25-oh-d)mentioning

confidence: 94%

Influence of Vitamin D2 Percentage on Accuracy of 4 Commercial Total 25-Hydroxyvitamin D Assays

et al. 2013

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The between-day CVs were between 5.1% and 12% at D 2 , D 3 , and total D concentrations of 12.9 -143 ng/mL (32.2-358 nmol/ L). The matrix effect was negligible. The linearity intervals for D 3 , D 2 , and total D were 3.0 -200 ng/mL (7.5-500 nmol/L), 1.0 -150 ng/mL (2.5-375 nmol/L), and 3.0 -350 ng/mL (7.5-875 nmol/L), respectively. The recoveries were 83.8%-109.2% (mean, 95.9%). The lower limits of quantification for D 3 and D 2 were 3.0 ng/mL (7.5 nmol/L) and 1.0 ng/mL (2.5 nmol/ L), respectively. We tested the 2011 College of American Pathologists proficiency testing set ABVD-01-05 with the assay, and all results were within Ϯ25% of target values (mean bias, Ϫ1.2%). Interference by 3-epi-25-OH-D 3 was negligible, because our study included only samples from adults (21-97 years) (3 ).We randomly chose samples from 149 adult patients submitted for 25-OH-D testing by the Abbott Architect assay. All samples were tested by the LC-MS/MS assay (63 contained D 2 ), and subsets of these samples were aliquoted for the fol- For the Liaison assay, the 2 scenarios yielded similar coefficients of determination; however, these values were different for all of the other assays. The regression slopes and r 2 values of the Architect and Cobas assays both de- Letters to the Editor1274 Clinical Chemistry 59:8 (2013) creased when D 2 -containing samples were included. For the Centaur assay, on the other hand, the regression slope increased, and the r 2 decreased. The mean (SD) changes in bias from scenario (a) to (b) for the 4 assays are as follows: Liaison assay, Ϫ17.4% (6.0%) to Ϫ19.2% (4.6%); Architect assay, 0.9% (5.2%) to Ϫ7.8% (4.2%); Cobas assay, Ϫ11.4% (11.9%) to Ϫ15.5% (7.0%); Centaur assay, Ϫ27.7% (7.0%) to Ϫ17.0% (8.0%).The effect of D 2 % on assay bias is illustrated in Fig. 1B. The Liaison assay had an insignificant bias change as D 2 % increased. The Architect and Cobas assays showed an increasingly negative bias with increasing D 2 %, whereas the negative bias of the Centaur assay decreased and changed to a positive bias.We calculated D 2 crossreactivity normalized to D 3 reactivity (see Fig. 1 legend). The values for each assay in Although D 3 is the preferred supplement form for treating vitamin D deficiency (4, 5 ), we have observed a high prevalence of patient samples containing substantial amounts of D 2 , and we have demonstrated the different effects of D 2 content on the accuracies of commercial assays. Therefore, correctly interpreting total 25-OH-D results and monitoring patient compliance requires that vitamin D status, type of supplementation, and measurement method be considered.

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Section: -Hydroxyvitamin D (25-oh-d)mentioning

confidence: 94%

Influence of Vitamin D2 Percentage on Accuracy of 4 Commercial Total 25-Hydroxyvitamin D Assays

et al. 2013

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“…It was shown to be easily resolved from 25(OH)D 3 by reversed-phase chromatography, however, and eluted much earlier than 25(OH)D 3 [22]. Finally, the 3-epimer species of 25(OH)D 3 , which has been shown to be present in serum samples of adult subjects at considerable levels [8,22,24], was also well resolved from its 25(OH)D 3 variant [19] and therefore did not constitute an interference in our LRMS and HRMS assays. Figure S1 (Supplementary Material) illustrates the separation of 25(OH)D 3 and 3-epi-25(OH)D 3 for a CLD patient sample, showing~5% contribution of the epimer relative to the 25(OH)D 3 signal.…”

Section: Hrms: Tof-ms Versus Tof-ms/msmentioning

confidence: 98%

“…Furthermore, additional complications arise from isomeric contributions, where the C-3 epimer of 25(OH)D 3 presents a particular challenge [2]. Whether or not the epimeric species are biologically relevant is currently unknown [7]; regardless, chromatographic separation is essential to avoid bias in the analysis [1,2,8,9].…”

Section: Introductionmentioning

confidence: 99%

Triple Quadrupole Versus High Resolution Quadrupole-Time-of-Flight Mass Spectrometry for Quantitative LC-MS/MS Analysis of 25-Hydroxyvitamin D in Human Serum

Geib

Sleno

Hall

et al. 2016

J. Am. Soc. Mass Spectrom.

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Abstract. We describe a systematic comparison of high and low resolution LC-MS/ MS assays for quantification of 25-hydroxyvitamin D 3 in human serum. Identical sample preparation, chromatography separations, electrospray ionization sources, precursor ion selection, and ion activation were used; the two assays differed only in the implemented final mass analyzer stage; viz. high resolution quadrupolequadrupole-time-of-flight (QqTOF) versus low resolution triple quadrupole instruments. The results were assessed against measured concentration levels from a routine clinical chemiluminescence immunoassay. Isobaric interferences prevented the simple use of TOF-MS spectra for extraction of accurate masses and necessitated the application of collision-induced dissociation on the QqTOF platform. The two mass spectrometry assays provided very similar analytical figures of merit, reflecting the lack of relevant isobaric interferences in the MS/MS domain, and were successfully applied to determine the levels of 25-hydroxyvitamin D for patients with chronic liver disease.

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“…The relevance of this finding is still elusive since probably in most individuals (except children) the concentrations of the 3-epi isomer is low and there is insufficient knowledge about the actual biological role of the compound; however, it is illustrated that the analytical specificity of LC-MS/MS is not absolute and must be questioned in a systematic way, as it is also the case with immunoassays. It further shows that delicate chromatographic separation -even including isomer separationmay be required for the accurate LC-MS/MS results (28).…”

Section: -Hydroxyvitamin D -An Exemplary Analytementioning

confidence: 99%

Vitamin D – challenges in diagnosing and monitoring of hypovitaminosis D / Vitamin D – izazovi u dijagnozi i praćenju hipovitaminoze D

Vogeser¹,

Seger²

2012

Journal of Medical Biochemistry

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Summary: During recent years, it has been recognized that sub-optimum vitamin D-status, often defined as decrease of PTH concentrations in response to supplementation of vitamin D, is a very widespread finding with potential health effects on a population level. As a consequence, there is a con tinuously increasing interest in the laboratory-based asses sment of the vitamin D status, with 25-hydroxyvitamin D as the most widely used analyte. However, there is a number of challenges in the characterization of the individual vitamin D status that are addressed in this article; they in clude analytical issues of 25-hydroxyvitamin D measurement; interpretation of results; development of guidelines for rational indication for laboratory testing; and evaluation of the role of 25-hydroxyvitamin D in the context of complementary markers of the vitamin D status. Keywords: vitamin D, hypovitaminosis D, reference ranges, analyticsKratak sadr`aj: Poslednjih godina je prime}eno da je suboptimalni status vitamina D, koji se ~esto defini{e smanjenjem koncentracije PTH kao odgovor na suplementaciju vitaminom D, veoma rasprostranjeno stanje sa potencijalnim efektima na zdravlje populacije. Kao posledica, interes za laboratorijsko odre|ivanje statusa vitamina D kontinuirano raste, gde se kao analit naj~e{}e koristi 25-hidroksivitamin D. Me|utim, postoji veliki broj izazova u karakterizaciji statusa vitamina D o kojima se govori u ovom radu; oni uklju~uju analiti~ke probleme u odre|ivanju 25-hidroksivitamina D; interpretaciju rezultata; definisanje preporuka za racionalnu indikaciju za laboratorijsko odre|ivanje; i procenu uloge 25-hidroksivitamina D u kontekstu komplementarnih markera statusa vitamina D.Klju~ne re~i: vitamin D, hipovitaminoza D, referentne vrednosti, analitika disorder in the first half of the last century was a major and very impressive success of scientifically based medicine. However, following the eradication of severe rickets due to widespread supplementation of vitamin D during the first year of live, the vitamin D system drew little attention in medicine during the subsequent decades. This was changed in 1980s when the measurement of 25-hydroxyvitamin D and PTH as valid and convenient markers of the vitamin D status became routinely available by ligand binding tests. It became evident that poor vitamin D status, described as lowering of PTH levels following the administration of vitamin D, is very common in many populations worldwide at least in the wintertime. Furthermore, there has been a growing evidence that vitamin D is of relevance in many physiological and potentially pathophysiological systems beyond calcium and phosphate homeostasis, leading to the concept of noncalcemic or even pleiotropic effects of

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Fast Separation of 25-Hydroxyvitamin D3 from 3-Epi-25-Hydroxyvitamin D3 in Human Serum by Liquid Chromatography–Tandem Mass Spectrometry: Variable Prevalence of 3-Epi-25-Hydroxyvitamin D3 in Infants, Children, and Adults

Cited by 50 publications

References 5 publications

Influence of Vitamin D2 Percentage on Accuracy of 4 Commercial Total 25-Hydroxyvitamin D Assays

Influence of Vitamin D2 Percentage on Accuracy of 4 Commercial Total 25-Hydroxyvitamin D Assays

Triple Quadrupole Versus High Resolution Quadrupole-Time-of-Flight Mass Spectrometry for Quantitative LC-MS/MS Analysis of 25-Hydroxyvitamin D in Human Serum

Vitamin D – challenges in diagnosing and monitoring of hypovitaminosis D / Vitamin D – izazovi u dijagnozi i praćenju hipovitaminoze D

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