On line dual microdialysis with ESI-MS for direct analysis of complex biological samples and microorganism lysates

被引:71
作者
Liu, CL
Hofstadler, SA
Bresson, JA
Udseth, HR
Tsukuda, T
Smith, RD
Snyder, AP
机构
[1] Pacific NW Lab, Environm Mol Sci Lab, Richland, WA 99352 USA
[2] USA, Edgewood Res, Ctr Dev & Engn, Aberdeen, Peoples R China
关键词
D O I
10.1021/ac971193k
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
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 mu M bovine serum albumin (BSA), 4.0 mu M cytochrome c, 2.3 mu M ubiquitin, and 9.4 mu M 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 similar to 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.
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页码:1797 / 1801
页数:5
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