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A new method for preparing disposable microsprayers, for on-line capillary-zone electrophoresis/electrospray mass spectrometry (CZE/ES-MS) at sub-microliter per minute flow rates, is described. Different means of coupling these microsprayers to the CZE columns are presented, and compared with the more traditional one-piece capillary arrangement used for on-line CZE/ES-MS analysis. Enhancement of separation performance was obtained using capillaries coated with (acryloylamino) propyltrimethylammonium chloride reagents, an attractive alternative to other amine-coated capillaries. The use of disposable microsprayers provided ease of operation and flexibility for on-line CZE/ES-MS experiments, since CZE columns can be prepared independently from the microsprayers. The application of these microsprayers to unattended CZE/ES-MS operation for more than 8 hours was evaluated, and found to provide relative standard deviation values of 3–8% on peak areas and less than 1% on migration times. Full-scan mass spectral acquisition was obtained for low femtomol injections of peptides. The technique was applied to the analysis of a single chain lectin from Phaseolus vulgaris, using both CZE/ES-MS and CZE combined with tandem mass spectrometry. The mass spectra data obtained in this way provided information on the structure of glycopeptides identified in digests of the glycoprotein. © 1997 Crown Copyright Canada
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