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Ion spray liquid chromatographic/mass spectrometric characterization of bile acids
Author(s) -
Warrack Bethanne M.,
DiDonato Gerald C.
Publication year - 1993
Publication title -
biological mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1052-9306
DOI - 10.1002/bms.1200220202
Subject(s) - chemistry , trifluoroacetic acid , chromatography , protonation , acetonitrile , mass spectrometry , adduct , bile acid , solvent , ion , organic chemistry , biochemistry
The ion spray mass spectra of selected bile acid standards are presented. Full‐scan positive ion spectra obtained in solutions of acetonitrile–water–trifluoroacetic acid and methanol–water–trifluoroacetic acid show the presence of intense [M + S + H] + and [M + 2S + H] + ions (where S is either methanol or acetonitrile, depending on the solvent system) in addition to [M + H] + ions. The relative abundance of the protonated and solvent adduct ions are reproducible and unique for several of the bile acids, and provide a means of identifying isomeric bile salts. Adduct ions are most abundant for free acids and weaker for glycine and taurine conjugates. On‐line ion spray liquid chromatography/mass spectrometry (LC/MS) using a 3 cm × 4.6 mm cartridge column containing 3 μm particle size C 18 packing was utilized for the separation and identification of components of monkey bile. Eight common bile acid conjugates were separated in less than 8 min utilizing a water–acetonitrile gradient with 0.1% trifluoroacetic acid as the mobile‐phase modifier. Full‐scan data were acquired from 1 μl of untreated bile injected on‐column. Identification of component bile acids from the equivalent of 1 nl injected on‐column was obtained with the use of a 1 mm diameter microbore column. Limits of detection for bile acid conjugates using a microbore column and selected ion monitoring (SIM) range from 40 to 100 fmol.