Justin Wiseman (Presenter)
Prosolia
Authorship: Jeffrey S. Patrick, Joe Kennedy, Justin Wiseman
Prosolia, Inc. Indianapolis, IN USA
| Short Abstract The relative distribution of metabolites and lipids in serum has been linked to a broad range of disease states, among them diabetes. The analysis of the samples typically requires various levels of sample preparation followed by time-consuming and costly analysis using HPLC-MS. A simple and informative screen could provide rapid information to make differential health determinations is discussed here. Serum samples from diabetic, fatty and control rats is analyzed using paper spray interfaced to high resolution mass spectrometry. The analysis is simple and uses positive and negative ion modes. For a small sample set (n=3 each for control, fatty and diabetic Zucker rats) significant differences in phosphatidyl and lysophosphatidyl cholines, ethanolamines, serines and other lipids, and in small molecule metabolites including fatty acids. The 2 minute analysis uses 15 mcl of sample. |
Long Abstract
The relative distribution of metabolites and lipids in serum has been linked to a broad range of disease states, among them diabetes. The value of this has been demonstrated broadly. The analysis of the samples typically requires various levels of sample preparation followed by time-consuming and costly analysis using HPLC-MS. Translating these analyses to clinical applications to monitor disease progression/status/health would be beneficial. To be useful this will require a substantial reduction in sample preparation and analysis time. New technologies are offering alternatives to traditional chromatography-based analyses. PaperSpray, in which a sample is deposited onto a porous substrate and analyzed directly by mass spectrometry, provides one such alternative. Here serum samples from diabetic, fatty and control rats are analyzed using paper spray. A small portion (8 microliters) of untreated plasma or serum is deposited onto a paper cartridge. Triplicate portions of each of 9 samples are analyzed. Analytes are eluted using a mixture of aqueous acetonitrile containing formic acid. The Paperspray unit (Velox 360) is interfaced to a QExactive Focus high resolution mass spectrometry. Full scan spectra are acquired for the sample. The spectra provide a wealth of information and include large lipids (TAGs; PCs; PSs; PAs; PIs and more), free fatty acids, amino acids, organic acids and others. Data are evaluated in different ways: 1) spectral differences within the populations are evaluated; 2) they are normalized and processed without filtering in XLStat and other statistical programs. The analysis is simple and uses positive and negative ion modes. For a small sample set (n=3 each for control, fatty and diabetic Zucker rats) significant differences in phosphatidyl and lysophosphatidyl cholines, ethanolamines, serines and other lipids, and in small molecule metabolites including fatty acids. The results will be compared with results from other analytical findings. The analysis uses approximately 10 mcl of sample and is complete in less than 2 minutes per sample. This study demonstrates the utility of paper spray interfaced to high resolution mass spectrometry to provide useful differential analysis in a rapid fashion amenable to routine clinical analysis.
References & Acknowledgements:
| Description | Y/N | Source |
| Grants | no | |
| Salary | yes | Prosolia |
| Board Member | yes | Prosolia |
| Stock | yes | Prosolia |
| Expenses | no |
IP Royalty: no
| Planning to mention or discuss specific products or technology of the company(ies) listed above: | yes |