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Porphyrinogen fragmentation profiles by ultra‐high‐performance liquid chromatography/electrospray ionisation tandem mass spectrometry
Authors:Christopher M. Benton  Chang Kee Lim  Caje Moniz  Donald J. L. Jones
Affiliation:1. Clinical Biochemistry, King's College Hospital, , London, SE5 9RS UK;2. Cancer Studies and Molecular Medicine, RKCSB, University of Leicester, , Leicester, LE2 7LX UK;3. MRC Bioanalytical Science Group, Department of Biological Sciences, Birkbeck, University of London, , London, WC1E 7HX UK
Abstract:
An ultra‐high‐performance liquid chromatography/electrospray ionisation tandem mass spectrometry system is described for the separation and characterisation of uroporphyrinogen, heptacarboxylic acid porphyrinogen, hexacarboxylic acid porphyrinogen, pentacarboxylic acid porphyrinogen and coproporphyrinogen. The separation was carried out on a 100 mm × 2.1 mm Thermo‐Hypersil BDS column (2.4 µm average particle size) by gradient elution with a mixture of acetonitrile, methanol and 1 mol/L aqueous ammonium acetate buffer, pH 5.16, as eluent. The fragmentation pattern of each compound was established by collision‐induced dissociation tandem mass spectrometry. The most characteristic fragmentation was ring opening at one of the four methylene bridges of the protonated porphyrinogen molecule followed by further cleavages of methylene bridges linking the four pyrrole rings at various points to give product ions with methylenepyrrolenine, methylene‐dipyrrolenine and methylene‐tripyrrolenine structures. Copyright © 2011 John Wiley & Sons, Ltd.
Keywords:
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