Novel Applications of Comprehensive Two-Dimensional Gas Chromatography Time-of-Flight Mass Spectrometry.

[摘要] Comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry (GC × GC-ToFMS) was used to analyze extracted metabolites from the INS-1 cell line and pyrolysis products of whole sediment samples from a 1 million year old sapropel sequence. The detection and identification of metabolites extracted from INS-1 cells can be used to better understand the biochemical effects of varying glucose concentrations on the cell. The analysis of whole sediment sapropel pyrolysates can be used to obtain paleoclimatic and paleoceanographic information. Twenty seven target metabolites were detected and identified in extracts from INS-1 cells incubated at 3 mM, 7 mM and 17 mM glucose with excellent reproducibility. The level of eleven metabolites demonstrated significant change with glucose and 73, 80, and 65 analytes of interest were detected at 3 mM, 7 mM and 17 mM glucose, respectively, using a metabolomics approach consisting of a Fisher Ratio analysis and pathway mapping in Metscape. From the analysis, 32 metabolic pathways were determined to be involved, 12 can be related to β-cell dysfunction, glucose stimulated insulin secretion or diabetes. These pathways can be futher investigated to gain a better understanding of the metabolism triggered by glucose stimulation.Thirty fatty acid methyl esters (FAMEs) were reproducibly identified in extracts of INS-1 cells stimulated at 0 mM, 0.5 mM, 10 mM and 20 mM glucose. The level of 24 FAMEs was determined to significantly decrease at 20 mM glucose. Seven FAME levels were also determined to change significantly at 0.5 mM and 10 mM glucose. This work doubles the number of previously identified FAMEs in similar cell sets and demonstrates the utility of GC × GC-ToFMS for such analysis.Pyroysis GC × GC-ToFMS of a whole sediment sapropel sequence resulted in the detection of alkanes, alkenes, furans, thiophenes, pyrroles, alkyl aromatics, phenols and methylketones. Paleoclimatic and paleoceanographic information, in agreement with previous data, was obtained without the use of time and solvent intensive extractions. This report demonstrates the first example of pyrolysis GC × GC-ToFMS on a sapropel sample as well as the potential for continued use of this technique in the analysis of whole sediment samples.

[发布机构] University of Michigan

[效力级别] GCxGC

[关键词] Gas Chromatography;GCxGC;Two-dimensional Gas Chromatgoraphy;INS-1 Cells;Sapropels;Chemistry;Science;Chemistry