Journal
ANALYTICAL CHEMISTRY
Volume 88, Issue 13, Pages 6812-6819Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.6b01284
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Funding
- Research Council of the University of Oklahoma Norman Campus
- American Society for Mass Spectrometry Research Award - Waters Corporation
- Oklahoma Center for the Advancement of Science and Technology [HR 14-152]
- National Institutes of Health [R01GM116116]
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A unique mass spectrometry (MS) method has been developed to determine the negatively charged species in live single cells using the positive ionization mode. The method utilizes dicationic ion-pairing compounds through the miniaturized multifunctional device, the single-probe, for reactive MS analysis of live single cells under ambient conditions. In this study, two dicationic reagents, 1,5p entanediyl-bis(1-butylpyrrolidinium) difluoride (C-5(bpyr)(2)F-2) and 1,3-propanediyl-bis- I I I (tripropylphosphonium) difluoride (C-3(triprp)(2)F-2), were added in the solvent and introduced into single cells to extract cellular contents for real-time MS analysis. The negatively charged (1 charged) cell metabolites, which form stable ion-pairs (1+ charged) with dicationic compounds (2+ charged), were detected in positive ionization mode with a greatly improved sensitivity. We have tentatively assigned 192 and 70 negatively charged common metabolites as adducts with (C-5(bpyr)(2)F-2) and (C-5(triprp)(2)F-2), respectively, in three separate SCMS experiments in the positive ion mode. The total number of tentatively assigned metabolites is 285 for C-5(bpyr)(2)F-2 and 143 for C-3(triprp)(2)F-2. In addition, the selectivity of dicationic compounds in the complex formation allows for the discrimination of overlapped ion peaks with low abundances. Tandem (MS/MS) analyses at the single cell level were conducted for selected adduct ions for molecular identification. The utilization of the dicationic compounds in the single-probe MS technique provides an effective approach to the detection of a broad range of metabolites at the single cell level.
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