Comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry detection: analysis of amino acid and organic acid trimethylsilyl derivatives, with application to the analysis of metabolites in rye grass samples.
First, standard mixtures of trimethylsilyl (TMS) derivatives of amino acid and organic acid are analyzed by comprehensive two-dimensional (2D) gas chromatography (GC) coupled to time-of-flight mass spectrometry (GC x GC/TOFMS) in order to illustrate important issues regarding application of the technique. Specifically of interest is the extent to which the peak capacity of the 2D separation space has been utilized and the procedure by which the derivative standards are identified in the 2D separations using the mass spectral information. The resulting 2D separation is found to make extensive use of the GC x GC separation space provided by the complementary stationary phases employed. Second, in order to demonstrate GC x GC/TOFMS on two real sample types, trimethylsilyl metabolite derivatives were analyzed from extracts of common lawn grass samples (i.e., perennial rye grass), as a means to provide insight into both the pre and post harvest physiology. Various chemical components in the two rye grass extract samples were found to either emerge or disappear in relation to the trauma response. For example, a significant difference in the peak for the TMS derivative of malic acid was found. The successful analysis of various components was readily facilitated by the 2D separation, while a 1D separation would have produced too much peak overlap, thus impeding the analysis. The importance of using a GC x GC separation approach for the analysis of complex samples, such as metabolite extracts, is therefore demonstrated. The real-time analysis capability of GC x GC/TOFMS for multidimensional metabolite analysis makes this technique well suited to the high-throughput analysis of metabolomic samples, especially compared to slower, stopped-flow type separation approaches.
Hope
Janiece L
JL
Department of Chemistry, Center for Process Analytical Chemistry, Box 351700, University of Washington, Seattle, WA 98195-1700, USA.
Prazen
Bryan J
BJ
Nilsson
Erik J
EJ
Lidstrom
Mary E
ME
Synovec
Robert E
RE