title: Nanospray 'taxation' and how to avoid it.
abstract: In mass spectrometric analysis with nanospray ionization, some analytes were found to appear in spectra with a delay of tens of minutes, while a few others could not be detected at all. The effect was found to be related to cation-exchange chromatography with negative charge on the glass surface, and with the most affected peptide or protein ions having strong localization of positive charge in blocks of two or more adjacent basic amino acid residues (e.g. melittin). The 'affinity' to the glass surface was studied with a peptide mixture and bovine serum albumin (BSA) tryptic digest solutions at sub-micromolar concentration. About 20% fewer tryptic peptides could be identified from spectra recorded with a glass nanospray capillary compared to those acquired with either conventional 1 microL/min electrospray or a quartz nanospray capillary. Protein identification studies are not likely to be seriously affected by this loss, but other protein applications, such as investigations of mutations or post-translational modifications, may suffer due to reduced sequence coverage. Ways to avoid losses of useful ions are discussed.
authors.affiliation: MDS SCIEX, 71 Four Valley Drive, Concord, L4K 4V8 Canada.
authors.forename: I V
authors.initials: IV
authors.lastname: Chernushevich
authors.forename: U
authors.initials: U
authors.lastname: Bahr
authors.forename: M
authors.initials: M
authors.lastname: Karas
mesh_terms: D000336:Aerosols / Q000032:analysis* / Q000737:chemistry; D016477:Artifacts; D002852:Chromatography, Ion Exchange / Q000379:methods*; D036103:Nanotechnology / Q000379:methods*; D010455:Peptides / Q000032:analysis* / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D000336:Aerosols; D010455:Peptides