Factors contributing to peak broadening and mass accuracy in the characterization of intact spores using matrix‐assisted laser desorption/ionization coupled with time‐of‐flight mass spectrometry

Factors contributing to peak broadening, accuracy and precision in mass assignment in the matrix-assisted laser desorption/ionization characterization of a lipopeptide desorbed from intact Bacillus spores were investigated. These spores were studied as an example of a thick, topologically irregular sample, which present a more difficult target than a pure peptide or protein. The type of matrix, matrix:sample ratio, laser fluence, and localized repetitive laser irradiation were all found to affect the full-width at half maximum of the biomarker. Both in-source and post-source phenomena were shown to contribute. Sample thickness had less effect. Precision and accuracy of mass assignment were also affected by matrix:sample ratio and laser fluence. In general, this sample was responsive to the same experimental variables as pure peptides, and the use of an internal standard produced significant improvements in precision and accuracy.