Skip to main content
SpringerLink
Log in

Separation and identification of monoglucuronides of vitamin D3 metabolites in urine by derivatization-assisted LC/ESI–MS/MS using a new Cookson-type reagent

  • Special Issue: Original Paper
  • Frontiers of Separation Analysis
  • Published:
Analytical Sciences Aims and scope Submit manuscript

Abstract

A liquid chromatography/electrospray ionization-tandem mass spectrometry (LC/ESI–MS/MS) method was developed using a new Cookson-type reagent, 4-[4-(1-pipelidinyl)phenyl]-1,2,4-triazoline-3,5-dione (PIPTAD), to analyze the monoglucuronides (Gs) of vitamin D3 metabolites in human urine. The G of 23S,25-dihydroxyvitamin D3 [23,25(OH)2D3] was previously found as a major metabolite of vitamin D3 in the urine, but its conjugation position remained undetermined. Determination of the position was an important research issue to clarify the whole picture of the excretion of surplus 25-hydroxyvitamin D3 [25(OH)D3, the circulating form of vitamin D3] in humans. After the pretreated urine sample was derivatized with PIPTAD, the peak corresponding to the G of 23,25(OH)2D3 was satisfactorily separated from the urine-derived interfering substances on reversed-phase LC, which could not be achieved by using the previous analogous reagent, DAPTAD. The PIPTAD-derivatized Gs of the vitamin D3 metabolites provided characteristic product ions useful for identifying the conjugation positions during the MS/MS. Accordingly, we successfully determined the glucuronidated position of 23,25(OH)2D3 to be the C23-hydroxy group. The developed method also enabled the simultaneous detection of Gs of 25(OH)D3 and 24R,25-dihydroxyvitamin D3 as well as 23,25(OH)2D3-23-G without interference from the urine components.

Graphical abstract

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Data availability

Data will be made available on request.

References

  1. A. Spiro, J.L. Buttriss, Nutr. Bull. 39, 322 (2014)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. V. Polzonetti, S. Pucciarelli, S. Vincenzetti, P. Polidori, Nutrients (2020). https://doi.org/10.3390/nu12061743

    Article  PubMed  PubMed Central  Google Scholar 

  3. T. Higashi, K. Shimada, T. Toyo’oka, J. Chromatogr. B 878, 1654 (2010)

    Article  CAS  Google Scholar 

  4. R.C. Tuckey, C.Y.S. Cheng, A.T. Slominski, J. Steroid Biochem. Mol. Biol. 186, 4 (2019)

    Article  CAS  PubMed  Google Scholar 

  5. J. Janoušek, V. Pilařová, K. Macáková, A. Nomura, J. Veiga-Matos, D.D.D. Silva, F. Remião, L. Saso, K. Malá-Ládová, J. Malý, L. Nováková, P. Mladěnka, Crit. Rev. Clin. Lab. Sci. 59, 517 (2022)

    Article  PubMed  Google Scholar 

  6. S. Ogawa, S. Ooki, K. Shinoda, T. Higashi, Anal. Bioanal. Chem. 406, 6647 (2014)

    Article  CAS  PubMed  Google Scholar 

  7. Y. Yoshimura, R. Hibi, A. Nakata, M. Togashi, S. Ogawa, T. Ishige, M. Satoh, F. Nomura, T. Higashi, Biomed. Chromatogr. (2019). https://doi.org/10.1002/bmc.4538

    Article  PubMed  Google Scholar 

  8. C. Gao, M.C. Bergagnini-Kolev, M.Z. Liao, Z. Wang, T. Wong, J.C. Calamia, Y.S. Lin, Q. Mao, K.E. Thummel, J. Chromatogr. B 1060, 158 (2017)

    Article  CAS  Google Scholar 

  9. C. Jenkinson, R. Desai, M.D. McLeod, J. Wolf Mueller, M. Hewison, D.J. Handelsman. J. Clin. Endocrinol. Metab. 107, 435 (2022)

    Article  PubMed  Google Scholar 

  10. Z. Wang, T. Wong, T. Hashizume, L.Z. Dickmann, M. Scian, N.J. Koszewski, J.P. Goff, R.L. Horst, A.S. Chaudhry, E.G. Schuetz, K.E. Thummel, Endocrinology 155, 2052 (2014)

    Article  PubMed  PubMed Central  Google Scholar 

  11. T. Wong, Z. Wang, B.D. Chapron, M. Suzuki, K.G. Claw, C. Gao, R.S. Foti, B. Prasad, A. Chapron, J. Calamia, A. Chaudhry, E.G. Schuetz, R.L. Horst, Q. Mao, I.H. de Boer, T.A. Thornton, K.E. Thummel, Drug Metab. Dispos. 46, 367 (2018)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. K. Takada, Y. Hagiwara, M. Togashi, A. Kittaka, F. Kawagoe, M. Uesugi, S. Nishimoto-Kusunose, T. Higashi, J. Steroid Biochem. Mol. Biol. (2022). https://doi.org/10.1016/j.jsbmb.2022.106133

    Article  PubMed  Google Scholar 

  13. T. Higashi, S. Homma, H. Iwata, K. Shimada, J. Pharm. Biomed. Anal. 29, 947 (2002)

    Article  CAS  PubMed  Google Scholar 

  14. T. Sakaki, N. Sawada, K. Komai, S. Shiozawa, S. Yamada, K. Yamamoto, Y. Ohyama, K. Inouye, Eur. J. Biochem. 267, 6158 (2000)

    Article  CAS  PubMed  Google Scholar 

  15. S. Ogawa, S. Ooki, M. Morohashi, K. Yamagata, T. Higashi, Rapid Commun. Mass Spectrom. 27, 2453 (2013)

    Article  CAS  PubMed  Google Scholar 

  16. W. Hobo, A. Jo, S. Koyagi, S. Otani, S. Ogawa, S. Nishimoto-Kusunose, T. Higashi, Chromatography 43, 111 (2022)

    Article  CAS  Google Scholar 

  17. K. Ohno, T. Hasegawa, T. Tamura, H. Utsumi, K. Yamashita, Anal. Sci. 34, 1017 (2018)

    Article  CAS  PubMed  Google Scholar 

  18. G.D. Carter, J.C. Jones, Ann. Clin. Biochem. 46, 79 (2009)

    Article  CAS  PubMed  Google Scholar 

  19. E.P.L. Hunter, S.G. Lias, J. Phys. Chem. Ref. Data 27, 413 (1998)

    Article  CAS  Google Scholar 

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

    CAS  Google Scholar 

  21. T. Higashi, K. Shimada, Biomed. Chromatogr. (2017). https://doi.org/10.1002/bmc.3808

    Article  PubMed  Google Scholar 

  22. T. Hayama, Anal. Sci. 36, 1151 (2020)

    Article  CAS  PubMed  Google Scholar 

  23. R. Thakare, Y.S. Chhonker, N. Gautam, J.A. Alamoudi, Y. Alnouti, J. Pharm. Biomed. Anal. 128, 426 (2016)

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This study was supported in part by the Japan Society for the Promotion of Science (JSPS) KAKENHI Grant Number 20K06990.

Author information

Authors and Affiliations

  1. Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba, 278-8510, Japan

    Kazunari Takada, Shoujiro Ogawa, Natsuki Yoshida & Tatsuya Higashi

  2. Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, 1 Sanzo, Gakuen‑cho, Fukuyama, 729‑0292, Japan

    Shoujiro Ogawa

Authors
  1. Kazunari Takada
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shoujiro Ogawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Natsuki Yoshida
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Tatsuya Higashi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Tatsuya Higashi.

Ethics declarations

Conflict of interest

The authors have declared no conflict of interest.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 329 KB)

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Takada, K., Ogawa, S., Yoshida, N. et al. Separation and identification of monoglucuronides of vitamin D3 metabolites in urine by derivatization-assisted LC/ESI–MS/MS using a new Cookson-type reagent. ANAL. SCI. 39, 1053–1063 (2023). https://doi.org/10.1007/s44211-023-00350-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s44211-023-00350-4

Keywords

Search

Navigation