An atmospheric pressure matrix-assisted laser desorption/ionization ion trap with enhanced sensitivity.
When atmospheric pressure matrix-assisted laser desorption/ionization (AP-MALDI) became commercially available, the technique generated a great deal of interest because ion production was decoupled from mass analysis. Mass accuracy and resolution were therefore dependent on parameters governing the mass analyzer rather than the matrix and sample preparation. Researchers have successfully used AP-MALDI sources with both orthogonal acceleration time-of-flight (oaTOFMS) and ion trap mass spectrometers. However, one limitation of the technique has been sensitivity, especially for mixtures of peptides generated from tryptic digests. In this work, data are presented documenting an increase in sensitivity of approximately two orders of magnitude as compared with results previously reported in the literature. The improvement in sensitivity is thought to derive primarily from the novel use of a countercurrent heated gas stream directed at the sample, although the target plate position and ion sampling configuration have also been optimized to reduce chemical noise from low molecular weight ions. A tryptic digest of BSA containing 125 attomoles on the plate was successfully identified in MS-only mode, while MS/MS analysis of 250 attomoles of the same digest provided product ion spectra with sufficient information to identify the protein. More complicated mixtures of standard proteins were used to model proteomics experiments, and preliminary data suggest a minimum working dynamic range of 20-fold for the analysis of mixtures of protein digests.
Miller
Christine A
CA
Agilent Technologies, 5301 Stevens Creek Blvd, Santa Clara, CA 95051, USA.
Yi
Donghui
D
Perkins
Patrick D
PD
D000818:Animals; D001274:Atmospheric Pressure; D002417:Cattle; D010446:Peptide Fragments / Q000032:analysis* / Q000737:chemistry; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000295:instrumentation* / Q000379:methods
D010446:Peptide Fragments; D012710:Serum Albumin, Bovine