A new sensitive LC/MS/MS analysis of Vitamin D metabolites using a click derivatization reagent, 2-nitrosopyridine

Debin Wan, Jun Yang, Bogdan Barnych, Sung Hee Hwang, Kin Sing Stephen Lee, Yongliang Cui, Jun Niu, Mitchell A. Watsky, Bruce D. Hammock

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

There is an increased demand for comprehensive analysis of Vitamin D metabolites. This is a major challenge, especially for 1α,25-dihydroxyVitamin D [1α,25(OH)2VitD], because it is biologically active at picomolar concentrations. 4-Phenyl-1,2,4-triazoline-3,5-dione (PTAD) was a revolutionary reagent in dramatically increasing sensitivity of all diene metabolites and allowing the routine analysis of the bioactive, but minor, Vitamin D metabolites. A second generation of reagents used large fixed charge groups that increased sensitivity at the cost of a deterioration in chromatographic separation of the Vitamin D derivatives. This precludes a survey of numerous Vitamin D metabolites without redesigning the chromatographic system used. 2-Nitrosopyridine (PyrNO) demonstrates that one can improve ionization and gain higher sensitivity over PTAD. The resulting Vitamin D derivatives facilitate high-resolution chromatographic separation of the major metabolites. Additionally, a liquid-liquid extraction followed by solid-phase extraction (LLE-SPE) was developed to selectively extract 1α,25(OH)2VitD, while reducing 2-to 4-fold ion suppression compared with SPE alone. LLE-SPE followed by PyrNO derivatization and LC/MS/MS analysis is a promising new method for quantifying Vitamin D metabolites in a smaller sample volume (100 μL of serum) than previously reported methods. The PyrNO derivatization method is based on the Diels-Alder reaction and thus is generally applicable to a variety diene analytes.

Original languageEnglish (US)
Pages (from-to)798-808
Number of pages11
JournalJournal of Lipid Research
Volume58
Issue number4
DOIs
StatePublished - 2017

Fingerprint

Metabolites
Vitamin D
Liquid-Liquid Extraction
Solid Phase Extraction
Liquids
Derivatives
Cycloaddition Reaction
Ionization
Deterioration
Ions
Serum

Keywords

  • 1,25-dihydroxyVitamin D
  • Major Vitamin D metabolites
  • Matrix effect
  • Quantification
  • Smaller sample volume

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

Cite this

Wan, D., Yang, J., Barnych, B., Hwang, S. H., Lee, K. S. S., Cui, Y., ... Hammock, B. D. (2017). A new sensitive LC/MS/MS analysis of Vitamin D metabolites using a click derivatization reagent, 2-nitrosopyridine. Journal of Lipid Research, 58(4), 798-808. https://doi.org/10.1194/jlr.D073536

A new sensitive LC/MS/MS analysis of Vitamin D metabolites using a click derivatization reagent, 2-nitrosopyridine. / Wan, Debin; Yang, Jun; Barnych, Bogdan; Hwang, Sung Hee; Lee, Kin Sing Stephen; Cui, Yongliang; Niu, Jun; Watsky, Mitchell A.; Hammock, Bruce D.

In: Journal of Lipid Research, Vol. 58, No. 4, 2017, p. 798-808.

Research output: Contribution to journalArticle

Wan, D, Yang, J, Barnych, B, Hwang, SH, Lee, KSS, Cui, Y, Niu, J, Watsky, MA & Hammock, BD 2017, 'A new sensitive LC/MS/MS analysis of Vitamin D metabolites using a click derivatization reagent, 2-nitrosopyridine', Journal of Lipid Research, vol. 58, no. 4, pp. 798-808. https://doi.org/10.1194/jlr.D073536
Wan, Debin ; Yang, Jun ; Barnych, Bogdan ; Hwang, Sung Hee ; Lee, Kin Sing Stephen ; Cui, Yongliang ; Niu, Jun ; Watsky, Mitchell A. ; Hammock, Bruce D. / A new sensitive LC/MS/MS analysis of Vitamin D metabolites using a click derivatization reagent, 2-nitrosopyridine. In: Journal of Lipid Research. 2017 ; Vol. 58, No. 4. pp. 798-808.
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