High-performance liquid chromatographic analysis of bile pigments as their native tetrapyrroles and as their dipyrrolic azosulfanilate derivatives

Gerard B. Odell, William S. Mogilevsky, Glenn R. Gourley

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

A reversed-phase high-performance liquid chromatographic (HPLC) analysis of bile pigments is described that provides baseline separation of the major bilirubin conjugates found in bile. The advantage of the technique is that the bile pigments can be analyzed directly as their native tetrapyrroles without prior solvent extractions or derivatization. The use of ammonium acetate in place of sodium salts permits preparative isolation and lyophilization of the pigments for mass spectroscopy. The derivatization of the pigments as their dipyrrolic azosulfanilates with subsequent HPLC analysis demonstrates baseline separation of the endo- and exovinyl azodipyrroles and allows identification of that half of the tetrapyrrole which contains the conjugate in the instances of monoglycosides.

Original languageEnglish (US)
Pages (from-to)287-298
Number of pages12
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Volume529
Issue numberC
DOIs
StatePublished - 1990

Bibliographical note

Funding Information:
These studies have been supported by NIH Research Grants DDK21668 (G.B.O),HD20914 (G.R.G.) andHS03352 (G.R.G.).

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