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Clinical utility of simultaneous quantitation of 25-hydroxyvitamin D and 24,25-dihydroxyvitamin D by LC-MS/MS involving derivatization with DMEQ-TAD.
J Clin Endocrinol Metab. 2014 Jul; 99(7):2567-74.JC

Abstract

CONTEXT

The discovery of hypercalcemic diseases due to loss-of-function mutations in 25-hydroxyvitamin D-24-hydroxylase has placed a new demand for sensitive and precise assays for 24,25-dihydroxyvitamin D [24,25-(OH)2D].

OBJECTIVE

We describe a novel liquid chromatography and tandem mass spectrometry-based method involving derivatization with DMEQ-TAD {4-[2-(6,7-dimethoxy-4-methyl-3,4-dihydroquinoxalinyl)ethyl]-1,2,4-triazoline-3,5-dione} to simultaneously assay multiple vitamin D metabolites including 25-hydroxyvitamin D (25-OH-D) and 24,25-(OH)2D using 100 μL of serum with a 5-minute run time.

DESIGN

The assay uses a newly synthesized internal standard d6-24,25-(OH)2D3 enabling the quantitation of 24,25-(OH)2D3 as well as the determination of the ratio of 25-OH-D3 to 24,25-(OH)2D3, a physiologically useful parameter.

SETTING

We report data on more than 1000 normal and disease samples involving vitamin D deficiency or hypercalcemia in addition to studies involving knockout mouse models.

RESULTS

The assay showed good correlation with samples from quality assurance schemes for 25-OH-D (25-OH-D2 and 25-OH-D3) determination (-2% to -5% bias) and exhibited low inter- and intraassay coefficients of variation (4%-7%) and lower limits of quantitation of 0.25-0.45 nmol/L. In clinical studies, we found a strong correlation between serum levels of 25-OH-D3 and 24,25-(OH)2D3 (r(2) = 0.80) in subjects over a broad range of 25-OH-D3 values and a marked lack of production of 24,25-(OH)2D3 below 25 nmol/L of 25-OH-D. The ratio of 25-OH-D3 to 24,25-(OH)2D3, which remained less than 25 in vitamin D-sufficient subjects (serum 25-OH-D < 50 nmol/L) but was greatly elevated (80-100) in patients with idiopathic infantile hypercalcemia.

CONCLUSIONS

The new method showed good utility in clinical settings involving vitamin D deficiency; supplementation with vitamin D and idiopathic infantile hypercalcemia, as well as in animal models with ablation of selected cytochrome P450-containing enzymes involved in vitamin D metabolism.

Authors+Show Affiliations

Department of Biomedical and Molecular Sciences (M.Ka., G.J.), Queen's University, Kingston, Ontario, Canada K7L 3N6; Bone Metabolism Unit (J.C.G.), Creighton University School of Medicine, Omaha, Nebraska 68131; Indiana University School of Medicine (M.P.), Indianapolis, Indiana 46202-5250; Department of General Pediatrics, (K-P. S., M.Ko.) University Children's Hospital, D-48149, Münster, Germany; Department of Biochemistry (H.F.D.), University of Wisconsin-Madison, Madison, Wisconsin 53706; Departmento de Quimica Organica (R.S., B.L., A.M., M.M.), Laboratorio Ignacio Ribas, University of Santiago de Compostela, 15782 Santiago de Compostela, Spain; McGill University and Shriners Hospital for Children-Canada (R.S.), Montréal, Québec, Canada H3G 1A6; Endocrine Unit (J.S.F.), Massachusetts General Hospital, Boston Massachusetts 02114; and Health Sciences Research, Waters Corporation (D.P.C), Stamford Avenue, Altrincham Rd, Wilmslow, SK9 4AX United Kingdom.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Evaluation Study
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

24670084

Citation

Kaufmann, Martin, et al. "Clinical Utility of Simultaneous Quantitation of 25-hydroxyvitamin D and 24,25-dihydroxyvitamin D By LC-MS/MS Involving Derivatization With DMEQ-TAD." The Journal of Clinical Endocrinology and Metabolism, vol. 99, no. 7, 2014, pp. 2567-74.
Kaufmann M, Gallagher JC, Peacock M, et al. Clinical utility of simultaneous quantitation of 25-hydroxyvitamin D and 24,25-dihydroxyvitamin D by LC-MS/MS involving derivatization with DMEQ-TAD. J Clin Endocrinol Metab. 2014;99(7):2567-74.
Kaufmann, M., Gallagher, J. C., Peacock, M., Schlingmann, K. P., Konrad, M., DeLuca, H. F., Sigueiro, R., Lopez, B., Mourino, A., Maestro, M., St-Arnaud, R., Finkelstein, J. S., Cooper, D. P., & Jones, G. (2014). Clinical utility of simultaneous quantitation of 25-hydroxyvitamin D and 24,25-dihydroxyvitamin D by LC-MS/MS involving derivatization with DMEQ-TAD. The Journal of Clinical Endocrinology and Metabolism, 99(7), 2567-74. https://doi.org/10.1210/jc.2013-4388
Kaufmann M, et al. Clinical Utility of Simultaneous Quantitation of 25-hydroxyvitamin D and 24,25-dihydroxyvitamin D By LC-MS/MS Involving Derivatization With DMEQ-TAD. J Clin Endocrinol Metab. 2014;99(7):2567-74. PubMed PMID: 24670084.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Clinical utility of simultaneous quantitation of 25-hydroxyvitamin D and 24,25-dihydroxyvitamin D by LC-MS/MS involving derivatization with DMEQ-TAD. AU - Kaufmann,Martin, AU - Gallagher,J Christopher, AU - Peacock,Munro, AU - Schlingmann,Karl-Peter, AU - Konrad,Martin, AU - DeLuca,Hector F, AU - Sigueiro,Rita, AU - Lopez,Borja, AU - Mourino,Antonio, AU - Maestro,Miguel, AU - St-Arnaud,René, AU - Finkelstein,Joel S, AU - Cooper,Donald P, AU - Jones,Glenville, Y1 - 2014/03/26/ PY - 2014/3/28/entrez PY - 2014/3/29/pubmed PY - 2014/9/30/medline SP - 2567 EP - 74 JF - The Journal of clinical endocrinology and metabolism JO - J Clin Endocrinol Metab VL - 99 IS - 7 N2 - CONTEXT: The discovery of hypercalcemic diseases due to loss-of-function mutations in 25-hydroxyvitamin D-24-hydroxylase has placed a new demand for sensitive and precise assays for 24,25-dihydroxyvitamin D [24,25-(OH)2D]. OBJECTIVE: We describe a novel liquid chromatography and tandem mass spectrometry-based method involving derivatization with DMEQ-TAD {4-[2-(6,7-dimethoxy-4-methyl-3,4-dihydroquinoxalinyl)ethyl]-1,2,4-triazoline-3,5-dione} to simultaneously assay multiple vitamin D metabolites including 25-hydroxyvitamin D (25-OH-D) and 24,25-(OH)2D using 100 μL of serum with a 5-minute run time. DESIGN: The assay uses a newly synthesized internal standard d6-24,25-(OH)2D3 enabling the quantitation of 24,25-(OH)2D3 as well as the determination of the ratio of 25-OH-D3 to 24,25-(OH)2D3, a physiologically useful parameter. SETTING: We report data on more than 1000 normal and disease samples involving vitamin D deficiency or hypercalcemia in addition to studies involving knockout mouse models. RESULTS: The assay showed good correlation with samples from quality assurance schemes for 25-OH-D (25-OH-D2 and 25-OH-D3) determination (-2% to -5% bias) and exhibited low inter- and intraassay coefficients of variation (4%-7%) and lower limits of quantitation of 0.25-0.45 nmol/L. In clinical studies, we found a strong correlation between serum levels of 25-OH-D3 and 24,25-(OH)2D3 (r(2) = 0.80) in subjects over a broad range of 25-OH-D3 values and a marked lack of production of 24,25-(OH)2D3 below 25 nmol/L of 25-OH-D. The ratio of 25-OH-D3 to 24,25-(OH)2D3, which remained less than 25 in vitamin D-sufficient subjects (serum 25-OH-D < 50 nmol/L) but was greatly elevated (80-100) in patients with idiopathic infantile hypercalcemia. CONCLUSIONS: The new method showed good utility in clinical settings involving vitamin D deficiency; supplementation with vitamin D and idiopathic infantile hypercalcemia, as well as in animal models with ablation of selected cytochrome P450-containing enzymes involved in vitamin D metabolism. SN - 1945-7197 UR - https://www.unboundmedicine.com/medline/citation/24670084/Clinical_utility_of_simultaneous_quantitation_of_25_hydroxyvitamin_D_and_2425_dihydroxyvitamin_D_by_LC_MS/MS_involving_derivatization_with_DMEQ_TAD_ L2 - https://academic.oup.com/jcem/article-lookup/doi/10.1210/jc.2013-4388 DB - PRIME DP - Unbound Medicine ER -