Evidence for a monoglucuronide of 1,25-dihydroxyvitamin D3 in rat bile

R. D. Litwiller, V. R. Mattox, I. Jardine, Rajiv Kumar

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Abstract

We have isolated and characterized a monoglucuronide fraction of 9,10-secocholesta-5, 7, 10(19)triene-1α, 3β, 25-triol, 5,6-cis isomer (1,25-dihydroxyvitamin D3) from rat bile. Polar radioactive metabolites of 1,25-dihydroxyvitamin D3 were purified by a sequence of chromatographic procedures which utilized Amberlite XAD-2, diethylaminohydroxypropyl Sephadex LH-20, liquid-liquid partition on paper, and reverse phase chromatography on C-18 microparticulate columns. A purified radioactive substance showed maximal absorbance at 264 nm, indicating the presence of a triene in the 5,6-cis configuration. Mass spectrometry by fast atom bombardment of the product demonstrated an ion at m/z 637 atomic mass units that is consistent with a natriated sodium salt of a monoglucuronide of 1,25-dihydroxyvitamin D3 ([MNa]Na+). Following methylation of the carboxylic acid group and formation of trimethylsilyl ethers of the hydroxyl groups, the fragmentation pattern of the product was compatible with that of a monoglucuronide of 1,25-dihydroxyvitamin D3. The intact metabolite was treated with β-glucuronidase and the aglycon was isolated by chromatography on microparticulate silica. The aglycon co-migrated with authentic 1,25-dihydroxyvitamin D3 during chromatography and it gave a mass fragmentation pattern consistent with 1,25-dihydroxyvitamin D3. The aglycon was bound by an intestinal cytosol receptor with essentially the same affinity as 1,25-dihydroxyvitamin D3. These findings indicate that bile contains a monoglucuronide of 1,25-dihydroxyvitamin D3.

Original languageEnglish (US)
Pages (from-to)7491-7494
Number of pages4
JournalJournal of Biological Chemistry
Volume257
Issue number13
StatePublished - 1982

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ASJC Scopus subject areas

  • Biochemistry

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Litwiller, R. D., Mattox, V. R., Jardine, I., & Kumar, R. (1982). Evidence for a monoglucuronide of 1,25-dihydroxyvitamin D3 in rat bile. Journal of Biological Chemistry, 257(13), 7491-7494.