ION SPRAY LIQUID-CHROMATOGRAPHY ION TRAP MASS-SPECTROMETRY DETERMINATION OF BIOMOLECULES

被引:112
作者
MCLUCKEY, SA [1 ]
VANBERKEL, GJ [1 ]
GLISH, GL [1 ]
HUANG, EC [1 ]
HENION, JD [1 ]
机构
[1] CORNELL UNIV,NEW YORK STATE COLL VET MED,DRUG TESTING & TOXICOL DIAGNOST LAB,ITHACA,NY 14850
关键词
D O I
10.1021/ac00004a015
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
The first on-line coupling of microbore high-performance liquid chromatography (HPLC) with an ion trap mass spectrometer equipped for sampling ions formed by electrospray ionization at atmospheric pressure is reported. A pneumatically assisted electrospray (ion spray) liquid chromatograph/mass spectrometer interface was used to couple reversed-phase, linear solvent gradient separations of biologically important compounds, including peptides and proteins, with the ion trap mass spectrometer. Initial gradient HPLC conditions consisting of 95% aqueous 0.15% trifluoroacetic acid in acetonitrile maintained at a flow of 40-mu-L/min were used with the system. Useful ion current profiles for levels down to 2.5 pmol per component of tryptic peptides and 0.3 pmol of injected human serum albumin (HSA) were obtained under these conditions. Full-scan mass spectra of multiply protonated molecules are shown for the target compounds at these levels. On-line HPLC/mass spectrometry (MS) molecular weight determinations for cytochrome c, HSA, and myoglobin are shown as well as LC/MS2 and LC/MS3 determinations of selected tryptic peptides in a protein tryptic digest. These applications demonstrate the analytical potential for the ion trap mass spectrometer combined with HPLC through its high detection sensitivity and MS(n) capability.
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页码:375 / 383
页数:9
相关论文
共 48 条
[1]  
[Anonymous], 1989, J MICROCOLUMN SEP, DOI DOI 10.1002/mcs.1220010105
[2]   MICRO AND MICROBORE LIQUID-CHROMATOGRAPHY FAST ATOM BOMBARDMENT-MASS SPECTROMETRY (LC-FAB-MS) ANALYSIS OF PEPTIDES AND ANTIBIOTICS [J].
ASHCROFT, AE .
ORGANIC MASS SPECTROMETRY, 1987, 22 (11) :754-757
[3]  
BARINAGA CJ, 1989, RAPID COMMUN MASS SP, V3, P160
[4]   DETERMINATION OF SULFONATED AZO DYES BY LIQUID-CHROMATOGRAPHY ATMOSPHERIC-PRESSURE IONIZATION MASS-SPECTROMETRY [J].
BRUINS, AP ;
WEIDOLF, LOG ;
HENION, JD ;
BUDDE, WL .
ANALYTICAL CHEMISTRY, 1987, 59 (22) :2647-2652
[5]   ION SPRAY INTERFACE FOR COMBINED LIQUID CHROMATOGRAPHY/ATMOSPHERIC PRESSURE IONIZATION MASS-SPECTROMETRY [J].
BRUINS, AP ;
COVEY, TR ;
HENION, JD .
ANALYTICAL CHEMISTRY, 1987, 59 (22) :2642-2646
[6]  
Busch KL., 1988, MASS SPECTROMETRY MA
[7]   CONTINUOUS-FLOW FAST ATOM - BOMBARDMENT MASS-SPECTROMETRY [J].
CAPRIOLI, RM .
ANALYTICAL CHEMISTRY, 1990, 62 (08) :A477-A487
[8]   ANALYSIS OF BIOCHEMICAL REACTIONS WITH MOLECULAR SPECIFICITY USING FAST ATOM BOMBARDMENT MASS-SPECTROMETRY [J].
CAPRIOLI, RM .
BIOCHEMISTRY, 1988, 27 (02) :513-521
[9]   ION CHROMATOGRAPHY MASS-SPECTROMETRY FOR THE DETERMINATION OF ORGANIC AMMONIUM AND SULFATE COMPOUNDS [J].
CONBOY, JJ ;
HENION, JD ;
MARTIN, MW ;
ZWEIGENBAUM, JA .
ANALYTICAL CHEMISTRY, 1990, 62 (08) :800-807
[10]  
Covey T R, 1988, Rapid Commun Mass Spectrom, V2, P249, DOI 10.1002/rcm.1290021111