Metabolic profiling of major vitamin D metabolites using Diels–Alder derivatization and ultra-performance liquid chromatography–tandem mass spectrometry

  • Pavel A. Aronov
  • Laura M. Hall
  • Katja Dettmer
  • Charles B. Stephensen
  • Bruce D. Hammock
Original Paper

Abstract

Biologically active forms of vitamin D are important analytical targets in both research and clinical practice. The current technology is such that each of the vitamin D metabolites is usually analyzed by individual assay. However, current LC-MS technologies allow the simultaneous metabolic profiling of entire biochemical pathways. The impediment to the metabolic profiling of vitamin D metabolites is the low level of 1α,25-dihydroxyvitamin D3 in human serum (15–60 pg/mL). Here, we demonstrate that liquid–liquid or solid-phase extraction of vitamin D metabolites in combination with Diels–Alder derivatization with the commercially available reagent 4-phenyl-1,2,4-triazoline-3,5-dione (PTAD) followed by ultra-performance liquid chromatography (UPLC)–electrospray/tandem mass spectrometry analysis provides rapid and simultaneous quantification of 1α,25-dihydroxyvitamin D3, 1α,25-dihydroxyvitamin D2, 24R,25-dihydroxyvitamin D3, 25-hydroxyvitamin D3 and 25-hydroxyvitamin D2 in 0.5 mL human serum at a lower limit of quantification of 25 pg/mL. Precision ranged from 1.6–4.8 % and 5–16 % for 25-hydroxyvitamin D3 and 1α,25-dihydroxyvitamin D3, respectively, using solid-phase extraction.

Keywords

1α,25-Dihydroxyvitamin D3 25-Hydroxyvitamin D3 24R,25-Dihydroxyvitamin D3 UPLC LC-MS Metabolic profiling Derivatization 

Supplementary material

216_2008_2095_MOESM1_ESM.pdf (186 kb)
ESM 1(PDF 185 kb)

References

  1. 1.
    Fiehn O (2002) Plant Mol Biol 48(1–2):155–171CrossRefGoogle Scholar
  2. 2.
    Dettmer K, Aronov PA, Hammock BD (2007) Mass Spectrom Rev 26(1):51–78CrossRefGoogle Scholar
  3. 3.
    Zerwekh JE (2004) Ann Clin Biochem 41(4):272–281CrossRefGoogle Scholar
  4. 4.
    Maunsell Z, Wright DJ, Rainbow SJ (2005) Clin Chem 51(9):1683–1690CrossRefGoogle Scholar
  5. 5.
    Vogeser M, Kyriatsoulis A, Huber E, Kobold U (2004) Clin Chem 50(8):1415–1417CrossRefGoogle Scholar
  6. 6.
    Higashi T, Awada D, Shimada K (2001) Biol Pharm Bull 24(7):738–743CrossRefGoogle Scholar
  7. 7.
    Bouillon R, Okamura WH, Norman AW (1995) Endocr Rev 16(2):200–257CrossRefGoogle Scholar
  8. 8.
    Prosser DE, Jones G (2004) Trends Biochem Sci 29(12):664–673CrossRefGoogle Scholar
  9. 9.
    Armas LA, Hollis BW, Heaney RP (2004) J Clin Endocrinol Metab 89(11):5387–5391CrossRefGoogle Scholar
  10. 10.
    Houghton LA, Vieth R (2006) Am J Clin Nutr 84(4):694–697Google Scholar
  11. 11.
    Avioli LV, Lee SW, McDonald JE, Lund J, DeLuca HF (1967) J Clin Invest 46(6):983–992Google Scholar
  12. 12.
    Norman AW (2006) Endocrinol 147(12):5542–5548CrossRefGoogle Scholar
  13. 13.
    Boyan BD, Sylvia VL, Dean DD, Del Toro F, Schwartz Z (2002) Crit Rev Oral Biol Med 13(2):143–154CrossRefGoogle Scholar
  14. 14.
    Schwartz Z, Ehland H, Sylvia VL, Larsson D, Hardin RR, Bingham V, Lopez D, Dean DD, Boyan BD (2002) Endocrinol 143(7):2775–2786CrossRefGoogle Scholar
  15. 15.
    Boyan BD, Sylvia VL, Dean DD, Schwartz Z (2001) Steroids 66(3–5):363–374CrossRefGoogle Scholar
  16. 16.
    Kissmeyer AM, Sonne K (2001) J Chromatogr A 935(1–2):93–103CrossRefGoogle Scholar
  17. 17.
    Higashi T, Awada D, Shimada K (2001) Biomed Chromatogr 15(2):133–140CrossRefGoogle Scholar
  18. 18.
    Higashi T, Yamauchi A, Shimada K (2003) Anal Sci 19(6):941–943CrossRefGoogle Scholar
  19. 19.
    Higashi T, Homma S, Iwata H, Shimada K (2002) J Pharm Biomed Anal 29(5):947–955CrossRefGoogle Scholar
  20. 20.
    Wilson SR, Tulchinsky ML, Wu Y (1993) Bioorg Med Chem Lett 3(9):1805–1808CrossRefGoogle Scholar
  21. 21.
    Aronov PA, Dettmer K, Christiansen JA, Cornel AJ, Hammock BD (2005) J Agric Food Chem 53(9):3306–3312CrossRefGoogle Scholar
  22. 22.
    Murao N, Ishigai M, Sekiguchi N, Takahashi T, Aso Y (2005) Anal Biochem 346(1):158–166CrossRefGoogle Scholar
  23. 23.
    Anon (2001) J Adolesc Health 29(3, Suppl 1):5–6Google Scholar
  24. 24.
    Rogers AS, Futterman DK, Moscicki AB, Wilson CM, Ellenberg J, Vermund SH (1998) J Adolesc Health 22(4):300–311CrossRefGoogle Scholar
  25. 25.
    Hollis BW (2005) Detection of vitamin D and its major metabolites. In: Feldman D, Pike JW, Glorieux FH (eds) Vitamin D. Elsevier, AmsterdamGoogle Scholar
  26. 26.
    Carter GD, Carter CR, Gunter E, Jones J, Jones G, Makin HL, Sufi S (2004) J Steroid Biochem Mol Biol 89–90(1–5):467–471CrossRefGoogle Scholar
  27. 27.
    Giorgianni F, Cappiello A, Beranova-Giorgianni S, Palma P, Trufelli H, Desiderio DM (2004) Anal Chem 76(23):7028–7038CrossRefGoogle Scholar
  28. 28.
    Astecker N, Reddy GS, Herzig G, Vorisek G, Schuster I (2000) Mol Cell Endocrinol 170(1–2):91–101CrossRefGoogle Scholar
  29. 29.
    Singh RJ, Taylor RL, Reddy GS, Grebe SK (2006) J Clin Endocrinol Metab 91(8):3055–3061CrossRefGoogle Scholar
  30. 30.
    Holick MF (2003) J Cell Biochem 88(2):296–307CrossRefGoogle Scholar
  31. 31.
    Stephensen CB, Marquis GS, Kruzich LA, Douglas SD, Aldrovandi GM, Wilson CM (2006) Am J Clin Nutr 83(5):1135–1141Google Scholar
  32. 32.
    Madeddu G, Spanu A, Solinas P, Calia GM, Lovigu C, Chessa F, Mannazzu M, Falchi A, Mura MS, Madeddu G (2004) Q J Nucl Med Mol Imaging 48(1):39–48Google Scholar
  33. 33.
    Cozzolino M, Vidal M, Arcidiacono MV, Tebas P, Yarasheski KE, Dusso AS (2003) Aids 17(4):513–520CrossRefGoogle Scholar
  34. 34.
    Delahunt JW, Romeril KE (1994) J Acquir Immune Defic Syndr 7(8):871–872Google Scholar
  35. 35.
    Ahmed B, Jaspan JB (1993) Am J Med Sci 306(5):313–316CrossRefGoogle Scholar
  36. 36.
    Adams JS, Fernandez M, Gacad MA, Gill PS, Endres DB, Rasheed S, Singer FR (1989) Blood 73(1):235–239Google Scholar
  37. 37.
    Aly ES, Baig M, Khanna D, Baumann MA (1999) Int J Clin Pract 53(3):227–228Google Scholar
  38. 38.
    Freeman NJ, Holik D (2003) J Clin Oncol 21(1):170–172Google Scholar
  39. 39.
    Barbour GL, Coburn JW, Slatopolsky E, Norman AW, Horst RL (1981) N Engl J Med 305(8):440–443Google Scholar
  40. 40.
    Barnes PF, Modlin RL, Bikle DD, Adams JS (1989) J Clin Invest 83(5):1527–1532CrossRefGoogle Scholar
  41. 41.
    Akeno N, Saikatsu S, Kawane T, Horiuchi N (1997) Endocrinol 138(6):2233–2240CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Pavel A. Aronov
    • 1
  • Laura M. Hall
    • 2
  • Katja Dettmer
    • 3
  • Charles B. Stephensen
    • 2
  • Bruce D. Hammock
    • 4
  1. 1.Department of EntomologyUniversity of CaliforniaDavisUSA
  2. 2.Department of Nutrition and USDA Western Human Nutrition Research CenterUniversity of CaliforniaDavisUSA
  3. 3.Institute of Functional GenomicsUniversity of RegensburgRegensburgGermany
  4. 4.Department of Entomology and U.C. Davis Cancer Research CenterUniversity of CaliforniaDavisUSA

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