Biomedical applications of high-performance liquid chromatography-mass spectrometry with continuous-flow fast atom bombardment.
This report describes the application of high-performance liquid chromatography combined with continuous-flow fast atom bombardment mass spectrometry to analytical problems in the biomedical laboratory. Applications include the compound-specific detection of diagnostic acylcarnitines in human urine, the separation and analysis of acyl-coenzyme A thioesters, and qualitative studies on complex mixtures of modified peptides (dansyl and dinitrophenyl derivatives). For each of these applications standard analytical columns (3.9 mm I.D.) and 1 ml/min flow-rates were employed with post-column stream splitting (1:100) before mass spectrometry. Various mobile phase compositions and solvent gradients were employed. The addition of 1-5% glycerol to the mobile phase was shown to have little effect on the chromatography. For all compounds studied (acylcarnitines, acyl-coenzyme A thioesters, and derivatized peptides) molecular weight information was obtained and sufficient sensitivity was achieved to allow unambiguous identification of trace components in complex mixtures.
Millington
D S
DS
Division of Pediatric Genetics and Metabolism, Duke University Medical Center, Durham, NC 27710.
Norwood
D L
DL
Kodo
N
N
Moore
R
R
Green
M D
MD
Berman
J
J
D000214:Acyl Coenzyme A / Q000032:analysis; D002331:Carnitine / Q000031:analogs & derivatives / Q000652:urine; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation*; D004140:Dinitrophenols / Q000032:analysis; D007202:Indicators and Reagents; D010455:Peptides / Q000032:analysis*; D012015:Reference Standards; D013050:Spectrometry, Fluorescence; D016339:Spectrometry, Mass, Fast Atom Bombardment / Q000295:instrumentation*; D013056:Spectrophotometry, Ultraviolet
D000214:Acyl Coenzyme A; D004140:Dinitrophenols; D007202:Indicators and Reagents; D010455:Peptides; D002331:Carnitine