Novel polymer composite to eliminate background matrix ions in matrix assisted laser desorption/ionization-mass spectrometry.
Matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) is rarely used for the analysis of small molecules (< 700 Da) because the low m/z signal is overwhelmed by a high background of matrix ions. We have developed a solution to this problem that employs a novel polymer composite which is formed by covalently cross-linking alpha-cyano-4-hydroxycinnamic acid (HCCA) to SU-8 photoresist via cationic photo-polymerization. Since the HCCA molecules are immobilized, background noise resulting from the matrix ions is significantly reduced or eliminated. Moreover, owing to the hydrophobic surface of the polymer film, the sample spots shrink during solvent evaporation and thus the analytes can be concentrated. As a result, this polymer composite improves detection sensitivity and extends the analyzable species to the low-mass region. The covalent incorporation of HCCA with SU-8 was validated with reflectance FTIR spectroscopy, and the polymer surface was characterized with scanning electron microscopy (SEM). Using MRFA, a small peptide as a standard, 8 mg of HCCA per mL of SU-8 photoresist was found to yield the highest sensitivity and the lowest background noise. Analytes such as peptides or small organic molecules were further examined on this composite surface and no analyte degradation was observed. In a trial of peptide mass fingerprinting of cytochrome c on the composite substrate, the inclusion of low m/z tryptic peptides in the database search dramatically improved the protein identification probability score.
Hua
Lin
L
Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672.
Low
Teck Yew
TY
Meng
Wei
W
Chan-Park
Mary B
MB
Sze
Siu Kwan
SK
D003373:Coumaric Acids; D008855:Microscopy, Electron, Scanning; D053758:Nanoparticles; D011108:Polymers; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D003373:Coumaric Acids; D011108:Polymers; C007175:alpha-cyano-4-hydroxycinnamate