Identification of conjugation positions of urinary glucuronidated vitamin D3 metabolites by LC/ESI-MS/MS after conversion to MS/MS-fragmentable derivatives.

A liquid chromatography/electrospray ionization-tandem mass spectrometry-based method was developed for the identification of the conjugation positions of the monoglucuronides of 25-hydroxyvitamin D3 [25(OH)D3 ] and 24,25-dihydroxyvitamin D3 [24,25(OH)2 D3 ] in human urine. The method employed derivatization with 4-(4-dimethylaminophenyl)-1,2,4-triazoline-3,5-dione to convert the glucuronides into fragmentable derivatives, which provided useful product ions for identifying the conjugation positions during the MS/MS. The derivatization also enhanced the assay sensitivity and specificity for urine sample analysis. The positional isomeric monoglucuronides, 25(OH)D3 -3- and -25-glucuronides, or 24,25(OH)2 D3 -3-, -24- and -25-glucuronides, were completely separated from each other under the optimized LC conditions. Using this method, the conjugation positions were successfully determined to be the C3 and C24 positions for the glucuronidated 25(OH)D3 and 24,25(OH)2 D3 , respectively. The 3-glucuronide was not present for 24,25(OH)2 D3 , unlike 25(OH)D3 , thus we found that selective glucuronidation occurs at the C24-hydroxy group for 24,25(OH)2 D3 .

• Publication year

  2019

• Venue

  Biomedical chromotography

• Publication date

  2019-07-01

• Fields of study

  Medicine, Chemistry

• Identifiers
  DOI 10.1002/bmc.4538PMID 30912169
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Polymorphic Human Sulfotransferase 2A1 Mediates the Formation of 25-Hydroxyvitamin D3-3-O-Sulfate, a Major Circulating Vitamin D Metabolite in Humans

  2018cited by this paper
• Hepatic Transport of 25-Hydroxyvitamin D3 Conjugates: A Mechanism of 25-Hydroxyvitamin D3 Delivery to the Intestinal Tract

  2018cited by this paper
• Sulfation of vitamin D3‐related compounds—identification and characterization of the responsible human cytosolic sulfotransferases

  2017cited by this paper
• Application of Cookson-type reagents for biomedical HPLC and LC/MS analyses: a brief overview.

  2017cited by this paper
• Simultaneous quantification of 25-hydroxyvitamin D3-3-sulfate and 25-hydroxyvitamin D3-3-glucuronide in human serum and plasma using liquid chromatography-tandem mass spectrometry coupled with DAPTAD-derivatization.

  2017cited by this paper
• Serum and urine vitamin D metabolite analysis in early preeclampsia

  2017cited by this paper
• A Method for Simultaneous Determination of 25-Hydroxyvitamin D3 and Its 3-Sulfate in Newborn Plasma by LC/ESI-MS/MS after Derivatization with a Proton-Affinitive Cookson-Type Reagent.

  2016cited by this paper
• Mass spectrometric profiling of vitamin D metabolites beyond 25-hydroxyvitamin D.

  2015cited by this paper
• Analysis of urinary vitamin D3 metabolites by liquid chromatography/tandem mass spectrometry with ESI-enhancing and stable isotope-coded derivatization

  2014cited by this paper
• Human UGT1A4 and UGT1A3 conjugate 25-hydroxyvitamin D3: metabolite structure, kinetics, inducibility, and interindividual variability.

  2014cited by this paper
• A novel Cookson-type reagent for enhancing sensitivity and specificity in assessment of infant vitamin D status using liquid chromatography/tandem mass spectrometry.

  2013cited by this paper
• Characterization of urinary metabolites of vitamin D(3) in man under physiological conditions using liquid chromatography-tandem mass spectrometry.

  2002cited by this paper
• Characterization of new conjugated metabolites in bile of rats administered 24,25-dihydroxyvitamin D(3) and 25-hydroxyvitamin D(3).

  2000cited by this paper
• In vitro and in vivo glucuronidation of 24,25-dihydroxyvitamin D3.

  1999cited by this paper
• Quantitative determination of 25-hydroxyvitamin D3 3-sulphate in human plasma using high performance liquid chromatography.

  1995cited by this paper
• Retention behavior of conjugated metabolites of vitamin D and related compounds in high-performance liquid chromatography.

  1994cited by this paper
• 25‐Hydroxyvitamin D3 3‐sulphate is a major circulating form of vitamin D in man

  1985cited by this paper

• Association between serum 25-hydroxyvitamin D concentrations and urinary vitamin D metabolite concentrations measured by the NLucVDR assay.

  2025cites this paper
• Rapid Determination of 25-Hydroxyvitamin D3 and 24,25-Dihydroxyvitamin D3 in Human Urine Based on Polystyrene/Graphene Oxide Packed-Fibers Solid-Phase Extraction Coupled With High-Performance Liquid Chromatography and Atmospheric Pressure Chemical Ionization Tandem Mass Spectrometry.

  2025cites this paper
• Determination of vitamin D3 conjugated metabolites: a complementary view on hydroxylated metabolites.

  2023cites this paper
• Separation and identification of monoglucuronides of vitamin D_3 metabolites in urine by derivatization-assisted LC/ESI–MS/MS using a new Cookson-type reagent

  2023cites this paper
• 23,25-Dihydroxyvitamin D3 is Liberated as a Major Vitamin D3 Metabolite in Human Urine After Treatment With β-glucuronidase: Quantitative Comparison With 24,25-dihydroxyvitamin D3 by LC/MS/MS.

  2022cites this paper
• Development of a LC-MS/MS method to measure serum 3-sulfate and 3-glucuronide 25-hydroxyvitamin D3 metabolites; comparisons to unconjugated 25OHD in pregnancy and polycystic ovary syndrome.

  2021cites this paper
• Analysis of vitamin D metabolic markers by mass spectrometry: Recent progress regarding the "gold standard" method and integration into clinical practice.

  2021cites this paper
• 3-Epi-25-hydroxyvitamin D3 is a poor substrate for SULT2A1: Analysis of its 3-sulfate in cord plasma and recombinant human SULT2A1 incubate.

  2020cites this paper