Particle beam liquid chromatography-mass spectrometry on a double-focusing sector instrument.
A commercially available particle beam interface developed by the Vestec Corporation has been coupled to a double-focusing high-resolution mass spectrometer (VG ZAB 2F) via a momentum separator. An ultraviolet detector was in-line with the interface and allowed monitoring of the eluting materials. With this instrumentation, complete electron-impact (EI) mass spectra on synthetic mixtures of steroids, nucleosides, and several derivatives of amino acids were recorded. Amongst these the EI mass spectra of 2,4-dinitrophenyl, 9-fluorenylmethoxycarbonyl and phenylthiohydantoin derivatives were obtained. The standard LC conditions used here (250 mm x 4.6 mm I.D., reversed-phase C18 column, 1.0 or 1.5 ml/min flow-rate, isocratic or gradient programming) allowed separation of the mixtures. The resulting mass spectra were compared with those obtained using a direct insertion probe. The agreement was excellent in most cases. Sensitivity measurements were performed on cholesterol and caffeine. A complete low-resolution mass spectrum can be obtained on 100 ng of cholesterol. Single-ion monitoring of the M+.of 200 pg of caffeine gave a 4:1 signal-to-noise ratio at m/z 194. Complete high-resolution mass spectra were obtained on 5 micrograms of cholesterol (loop injection) and on every peak of a five-steroid mixture of 5 micrograms each. The accuracy of the mass measurements were better than 5 m.m.u. for most cases. Three to four scans could be obtained for each liquid chromatographic peak. Mass-analyzed ion kinetic energy spectra were recorded on the M+.of 2.5 micrograms of cholesterol at m/z 386. Similarly the B/E linked scan of the same ion was recorded on 2.5 micrograms and 500 ng.
Baczynskyj
L
L
Physical and Analytical Chemistry Research, Upjohn Company, Kalamazoo, MI 49001.
D000596:Amino Acids / Q000032:analysis; D002784:Cholesterol / Q000032:analysis; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation*; D004563:Electrochemistry; D007202:Indicators and Reagents; D013058:Mass Spectrometry / Q000295:instrumentation*; D000432:Methanol; D010455:Peptides / Q000032:analysis; D010669:Phenylthiohydantoin / Q000031:analogs & derivatives / Q000032:analysis; D013056:Spectrophotometry, Ultraviolet; D013256:Steroids / Q000032:analysis
D000596:Amino Acids; D007202:Indicators and Reagents; D010455:Peptides; D013256:Steroids; D010669:Phenylthiohydantoin; D002784:Cholesterol; D000432:Methanol