Skip to main content

Advertisement

SpringerLink
Log in

Investigation of C-3 Epimerization Mechanism of 24,25-Dihydroxyvitamin D3 in Rat Using Liquid Chromatography/Mass Spectrometry

  • Published:
Analytical Sciences Aims and scope Submit manuscript

Abstract

Studies on C-3 epimerization of 24,25-dihydroxyvitamin D3 [24,25(OH)2D3] were performed. 3-Epi-24,25(OH)2D3 was found to circulate in the blood of rats administered 24,25(OH)2D3, which was confirmed in comparison with a standard sample based on its chromatographic behavior during inclusion high-performance liquid chromatography using γ-cyclodextrin as a mobile phase additive and data obtained from liquid chromatography (LC)/mass spectrometry (MS) and gas chromatography/MS. The C-3 epimerization mechanism was investigated in in vitro experiments using [3α-2H]-24,25(OH)2D3. The disappearance of deuterium was monitored by LC/MS. As a result, it was clarified that the 3-epimer was formed via the 3-oxo-form as an intermediate. This epimerization was also found to be controlled by the enzyme existing in the liver cytosol fraction and to require NAD and NADPH as coenzymes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. T. Nakamura, T. Kurokawa, and H. Orimo, Calcif. Tissue Int., 1988, 43, 235.

    Article  CAS  Google Scholar 

  2. T. Nakamura, T. Kurokawa, and H. Orimo, Bone. 1989, 10, 117.

    Article  CAS  Google Scholar 

  3. M. J. Beckman, P. Tadikonda, E. Werner, J. Prahl, S. Yamada, and H. F. DeLuca, Biochemistry. 1996, 35, 8465.

    Article  CAS  Google Scholar 

  4. T. Higashi, R. Kikuchi, K. Miura, K. Shimada, H. Hiyamizu, H. Ooi, Y. Iwabuchi, S. Hatakeyama, and N. Kubodera, Biol. Pharm. Bull., 1999, 22, 767.

    Article  CAS  Google Scholar 

  5. T. Higashi, K. Miura, R. Kikuchi, K. Shimada, H. Hiyamizu, H. Ooi, Y. Iwabuchi, S. Hatakeyama, and N. Kubodera, Steroids. 2000, 65, 281.

    Article  CAS  Google Scholar 

  6. G. S. Reddy, M.-L. Siu-Caldera, I. Schuster, N. Astecker, K.-Y. Tserng, K. R. Muralidharan, W. H. Okamura, J. A. McLane, and M. R. Us Ko Kovic, “Vitamin D: Chemistry, Biology and Clinical Applications of the Steroid Hormone”, ed. A. W. Norman, R. Bouillon and M. Thomasset, 1997. Vitamin D Workshop, Inc., California, 139.

  7. S. Masuda, T. Okano, M. Kamao, and T. Kobayashi, “Vitamin D: Chemistry, Biology and Clinical Applications of the Steroid Hormone.#x201D;, ed. A. W. Norman, R. Bouillon, and M. Thomasset, 1997. Vitamin D Workshop, Inc., California, 159.

  8. H. Sekimoto, M.-L. Siu-Caldera, A. Weiskopf, P. Vouros, K. R. Muralidharan, W. H. Okamura, M. R. Us Ko Kovic´, and G. S. Reddy, FEBS Lett., 1999, 241, 278.

    Article  Google Scholar 

  9. R. C. Cookson, S. S. Gupte, I. D. R. Stevens, and C. T. Watts, Org. Synth., 1971, 51, 121.

    Article  CAS  Google Scholar 

  10. O. H. Lowry, N. J. Rosebrough, A. L. Farr, and R. J. Randall, J. Biol. Chem., 1951, 193, 265.

    Article  CAS  Google Scholar 

  11. E. Schrötter, B. Schönecker, U. Hauschild, P. Droescher, and H. Schick, Synthesis. 1990, 193.

    Google Scholar 

  12. T. Higashi, M. Horike, R. Kikuchi, and K. Shimada, Steroids. 1999, 64, 715.

    Article  CAS  Google Scholar 

  13. K. Shimada, Y. Hirose, and T. Oe, J. Liq. Chromatogr., 1989, 12, 1949.

    CAS  Google Scholar 

  14. K. Shimada, T. Oe, and M. Suzuki, J. Chromatogr., 1991, 558, 306.

    Article  CAS  Google Scholar 

  15. M. Shimizu and S. Yamada, Vitamins (Japan). 1994, 68, 15.

    CAS  Google Scholar 

  16. Private communication, M. Shimizu (Tokyo Medical and Dental University).

Download references

Author information

Authors and Affiliations

  1. Faculty of Pharmaceutical Sciences, Kanazawa University, Takara-machi, Kanazawa, 920-0934, Japan

    Tatsuya Higashi, Aki Ogasawara & Kazutake Shimada

Authors
  1. Tatsuya Higashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Aki Ogasawara
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kazutake Shimada
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Higashi, T., Ogasawara, A. & Shimada, K. Investigation of C-3 Epimerization Mechanism of 24,25-Dihydroxyvitamin D3 in Rat Using Liquid Chromatography/Mass Spectrometry. ANAL. SCI. 16, 477–482 (2000). https://doi.org/10.2116/analsci.16.477

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.2116/analsci.16.477

Search

Navigation