On-Line Dual Microdialysis with ESI-MS for Direct Analysis of Complex Biological Samples and Microorganism Lysates

A novel dual-microdialysis approach has been developed for fast and efficient fractionation and cleanup for ESI-MS and ESI MS/MS analyses of biological samples. A modified dynamic microdialyzer utilizing two mirror-image serpentine channels, which sandwich a regenerated cellulose membrane of selecte...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 1998-05, Vol.70 (9), p.1797-1801
Main Authors: Liu, Chuanliang, Hofstadler, Steven A, Bresson, James A, Udseth, Harold R, Tsukuda, Toyoko, Smith, Richard D, Snyder, A. Peter
Format: Article
Language:English
Subjects:
Analysis
Analytical chemistry
Bacteriology
Biological and medical sciences
Chemistry
Escherichia coli - chemistry
Exact sciences and technology
Fundamental and applied biological sciences. Psychology
General, instrumentation
Macromolecular Substances
Mass Spectrometry
Membranes
Microbiology
Microdialysis - instrumentation
Microdialysis - methods
Microorganisms
Molecular biology
Morphology, structure, chemical composition
Organic chemistry
Scientific imaging
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Summary:A novel dual-microdialysis approach has been developed for fast and efficient fractionation and cleanup for ESI-MS and ESI MS/MS analyses of biological samples. A modified dynamic microdialyzer utilizing two mirror-image serpentine channels, which sandwich a regenerated cellulose membrane of selected molecular weight cutoff, serves as the first stage for the removal of high-molecular-weight components and cellular residue. The second stage employs a hollow microdialysis capillary to remove low-molecular-weight species (e.g., salts) which can degrade or preclude analysis ESI-MS. A protein mixture consisting of 30 μM bovine serum albumin (BSA), 4.0 μM cytochrome c, 2.3 μM ubiquitin, and 9.4 μM bradykinin in 0.5 M NaCl was used to evaluate the performance of this system. Essentially complete removal of both BSA and NaCl was achieved, resulting in high-quality mass spectra containing only the lower molecular weight proteins. After passing through the on-line dual-microdialysis system, a crude bacteria cell lysate yielded clean ESI-mass spectra in ∼20 min. MS/MS of selected ions demonstrated abundant fragment ions and provided a second-dimension “fingerprint” of the complex cellular fraction. Preliminary application of this technique for direct characterization of microorganism lysates is presented.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac971193k