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Received on December 6, 2005
Accepted on March 8, 2006
Endocrinology and Metabolism |
1 Toxicology Laboratory, Department of Laboratory Medicine, Veterans' Affairs Medical Center, University of Wisconsin Hospital and Clinics, Madison, WI
2 Osteoporosis Clinical Center and Research Program, Department of Medicine, Veterans' Affairs Medical Center, University of Wisconsin Hospital and Clinics, Madison, WI
3 Osteoporosis Clinical Center and Research Program, Department of Medicine, and Geriatric Research, Education and Clinical Center, Veterans' Affairs Medical Center, University of Wisconsin Hospital and Clinics, Madison, WI
* To whom correspondence should be addressed. E-mail: gl.lensmeyer{at}hosp.wisc.edu.
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 AdvantageTM 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 µg/L to at least 200 µg/L. Comparison data were as follows: for HPLC vs LC-MS/MS, y = 1.02x - 1.81 µg/L (Sy|x = 2.13 µg/L; r = 0.998) for 25(OH)D3, and y = 0.903x - 0.596 µg/L (Sy|x = 2.56 µg/L; r = 0.9965) for 25(OH)D2; for HPLC vs Diasorin RIA, y = 0.709x - 5.86 µg/L (Sy|x = 7.35 µg/L; r = 0.7509); and for HPLC vs Nichols Advantage CPBA, y = 1.00x - 3.60 µg/L (Sy|x = 32.7 µg/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.
The following articles in journals at HighWire Press have cited this article:
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  V. B. Popat, K. A. Calis, V. H. Vanderhoof, G. Cizza, J. C. Reynolds, N. Sebring, J. F. Troendle, and L. M. Nelson Bone Mineral Density in Estrogen-Deficient Young Women J. Clin. Endocrinol. Metab., July 1, 2009; 94(7): 2277 - 2283. [Abstract] [Full Text] [PDF]
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 A. W Norman From vitamin D to hormone D: fundamentals of the vitamin D endocrine system essential for good health Am. J. Clinical Nutrition, August 1, 2008; 88(2): 491S - 499S. [Abstract] [Full Text] [PDF]
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K. W Phinney Development of a standard reference material for vitamin D in serum Am. J. Clinical Nutrition, August 1, 2008; 88(2): 511S - 512S. [Abstract] [Full Text] [PDF]
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 A. J. Rovner and K. O. O'Brien Hypovitaminosis D Among Healthy Children in the United States: A Review of the Current Evidence Arch Pediatr Adolesc Med, June 1, 2008; 162(6): 513 - 519. [Abstract] [Full Text] [PDF]
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N. Binkley, D. Krueger, D. Gemar, and M. K. Drezner Correlation among 25-Hydroxy-Vitamin D Assays J. Clin. Endocrinol. Metab., May 1, 2008; 93(5): 1804 - 1808. [Abstract] [Full Text] [PDF]
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 H. J. Roth, H. Schmidt-Gayk, H. Weber, and C. Niederau Accuracy and clinical implications of seven 25-hydroxyvitamin D methods compared with liquid chromatography-tandem mass spectrometry as a reference Ann Clin Biochem, March 1, 2008; 45(2): 153 - 159. [Abstract] [Full Text] [PDF]
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 P. F. Lata and M. E. Elliott Patient Assessment in the Diagnosis, Prevention, and Treatment of Osteoporosis Nutr Clin Pract, June 1, 2007; 22(3): 261 - 275. [Abstract] [Full Text] [PDF]
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N. Binkley, R. Novotny, D. Krueger, T. Kawahara, Y. G. Daida, G. Lensmeyer, B. W. Hollis, and M. K. Drezner Low Vitamin D Status despite Abundant Sun Exposure J. Clin. Endocrinol. Metab., June 1, 2007; 92(6): 2130 - 2135. [Abstract] [Full Text] [PDF]
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 J. A. Schmidt Measurement of 25-Hydroxyvitamin D Revisited Clin. Chem., December 1, 2006; 52(12): 2304 - 2305. [Full Text] [PDF]
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G. Lensmeyer, D. Wiebe, N. Binkley, and M. Drezner The authors of the article cited above respond: Clin. Chem., December 1, 2006; 52(12): 2305 - 2306. [Full Text] [PDF]
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N. Binkley, M. K. Drezner, and B. W. Hollis Laboratory reporting of 25-hydroxyvitamin d results: potential for clinical misinterpretation. Clin. Chem., November 1, 2006; 52(11): 2124 - 2125. [Full Text] [PDF]
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