An automated noncontact deposition interface for liquid chromatography matrix-assisted laser desorption/ionization mass spectrometry.
A new multichannel deposition system was developed for off-line liquid chromatography/matrix-assisted laser desorption/ionization mass spectrometry (LC/MALDI-MS). This system employs a pulsed electric field to transfer the eluents from multiple parallel columns directly onto MALDI targets without the column outlets touching the target surface. The deposition device performs well with a wide variety of solvents that have different viscosities, vapor pressures, polarities, and ionic strengths. Surface-modified targets were used to facilitate concentration and precise positioning of samples, allowing for efficient automation of high-throughput MALDI analysis. The operational properties of this system allow the user to prepare samples using MALDI matrixes whose properties range from hydrophilic to hydrophobic. The latter, exemplified by alpha-cyano-4-hydroxycinnamic acid, were typically processed with a multistep deposition method consisting of precoating of individual spots on the target plate, sample deposition, and sample recrystallization steps. Using this method, 50 amol of angiotensin II was detected reproducibly with high signal-to-noise ratio after LC separation. Experimental results show that there is no significant decrease in chromatographic resolution using this device. To assess the behavior of the apparatus for complex mixtures, 5 microg of a tryptic digest of the cytosolic proteins of yeast was analyzed by LC/MALDI-MS and more than 13,500 unique analytes were detected in a single LC/MS analysis.
Ericson
Christer
C
Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, California 92121, USA.
Phung
Qui T
QT
Horn
David M
DM
Peters
Eric C
EC
Fitchett
Jonathan R
JR
Ficarro
Scott B
SB
Salomon
Arthur R
AR
Brill
Laurence M
LM
Brock
Ansgar
A
D000804:Angiotensin II / Q000032:analysis; D001331:Automation; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D003600:Cytosol / Q000737:chemistry; D011506:Proteins / Q000032:analysis; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014357:Trypsin / Q000378:metabolism; D015003:Yeasts / Q000737:chemistry
D011506:Proteins; D000804:Angiotensin II; D014357:Trypsin