The impact of buffers and surfactants from micellar electrokinetic chromatography on matrix-assisted laser desorption ionization (MALDI) mass spectrometry of peptides. Effect of buffer type and concentration on mass determination by MALDI-time-of-flight mass spectrometry.
This paper describes the effect of various buffers, surfactants, and organic additives commonly encountered in capillary zone electrophoresis and micellar electrokinetic chromatography on the molecular weight determination of peptides by matrix-assisted laser desorption/ionization (MALDI) time-of-flight mass spectrometry. Signal-to-noise ratio generally decreased with increasing buffer concentration without affecting mass accuracy, but the type of buffer was also important. Good spectra were obtained with an ammonium acetate buffer up to a concentration of 500 mM without impacting ionization of either peptides or other mobile phase constituents. Ionization of organic additives, such as anionic surfactants, non-ionic surfactants, and cyclodextrins was buffer dependent and presented a problem when the mass of the additive was in the range of the peptide mass. Brij-35, Tween-80, and cyclodextrins all produced prominent spectra of their own in the presence of sodium or potassium containing buffers, but not with ammonium acetate. Cationization of these neutral species with sodium or potassium ions allowed them to acquire a positive charge and produce spectra. In contrast, the ammonium ion appears to be a poor cationizating agent. Ionization of neutral surfactants was suppressed in ammonium acetate without impacting the spectra of peptides. Ammonium acetate buffers containing 30 mM sodium dodecyl phosphate also gave spectra with good signal intensity and no interference from the surfactant. Suppression of peptide ionization in MALDI was a problem when methanol, tetrabutyl amine, or poly(vinyl alcohol) were used with either ammonium acetate, sodium phosphate, and N-(2-hydroxyethyl)piperazine-N-(2-ethansulfonic acid).
Amini
A
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Department of Chemistry, Purdue University, West Lafayette, IN 47907-1393, USA.
Dormady
S J
SJ
Riggs
L
L
Regnier
F E
FE
D002021:Buffers; D020374:Chromatography, Micellar Electrokinetic Capillary / Q000379:methods*; D008970:Molecular Weight; D010455:Peptides / Q000032:analysis*; D012492:Salts; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013501:Surface-Active Agents / Q000737:chemistry*
D002021:Buffers; D010455:Peptides; D012492:Salts; D013501:Surface-Active Agents