Effect of solvent composition on signal intensity in liquid chromatography-matrix-assisted laser desorption ionization experiments.
The use of reversed phase liquid chromatography for the preparation of complex peptide mixtures for analysis by matrix assisted laser desorption ionization mass spectrometry has led to the observation of the critical importance of the matrix/analyte formulation in regards to the percent organic solvent in the mixture. This paper outlines the study using liquid chromatography to systematically vary the acetonitrile concentration in the formulation used for MALDI spot preparation to examine the impact the parameter has on analyte signal intensity. The results show that for five of six peptides tested across a wide mass range a formulation of approximately 75% acetonitrile is optimal for average MALDI signal intensity as determined on both time-of-flight and quadropole mass spectrometers. Examination of the individual spots shows that the organic solvent content in formulation significantly affects parameters such as crystal density and morphology.
Hattan
Stephen J
SJ
Applied Biosystems, Discovery Proteomics and Small Molecule Research Center, 500 Old Connecticut Path, Framingham, MA 01701, USA. hattansn@appliedbiosystems.com
Marchese
Jason
J
Albertinetti
Michael
M
Krishnan
Srinivasan
S
Khainovski
Nikita
N
Juhasz
Peter
P
D002853:Chromatography, Liquid / Q000379:methods*; D012997:Solvents / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D012997:Solvents