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-mu-g of cholesterol (loop injection) and on every peak of a five-steroid mixture of 5-mu-g 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-mu-g of cholesterol at m/z 386. Similarly the B/E linked scan of the same ion was recorded on 2.5-mu-g and 500 ng.