Gas chromatography ms analysis

Endometrial tumors were analyzed from the four groups (N = 5/group) by Metabolon (Durham, NC) according to their standard protocols [30 (link), 31 (link)]. Briefly, unbiased global metabolomic profiling was achieved using methanol extracts of tumor tissues normalized to tissue weight. Analysis of extracts consisted of either ultrahigh performance liquid chromatography (Waters Corporation, Milford, MA) coupled with tandem mass spectrometry (UHPLC/MS/MS; Thermo-Finnigan, San Jose CA) in positive and negative ionization modes, or via gas chromatography/MS analysis (Thermo-Finnigan). Metabolites in tumor tissues were positively identified by matching chromatographic retention time, mass and MS/MS fragmentation patterns to a reference library of over 2500 purified, authenticated biochemicals. Identification of known chemical entities was based on comparison to metabolomic library entries of purified standards based on chromatographic properties and mass spectra. Data are presented as relative measures of “scaled intensity” and median scaling to 1. Missing values were imputed with the minimum.

Metabolomic profiling was performed on serum obtained from patients pre- and postmetformin treatment as well as on endometrial tumors obtained post-treatment at surgery. Samples were analyzed by Metabolon (Research Triangle Park, NC) according to their standard protocols 21 (link)–24 (link). Briefly, unbiased global metabolomic profiling was achieved using methanol extracts of serum or tumor tissues normalized to serum volume or tissue weight. Analysis of extracts consisted of either ultrahigh performance liquid chromatography (Waters Corporation, Milford, MA) coupled with tandem mass spectrometry (UHPLC/MS/MS; Thermo-Finnigan, San Jose CA) in positive and negative ionization modes, or via gas chromatography/MS analysis (Thermo-Finnigan). Metabolites in serum or tumor tissues were positively identified by matching chromatographic retention time, mass, and MS/MS fragmentation patterns to a reference library of over 2500 purified, authenticated biochemicals. Data are presented as relative measures of “scaled intensity” and median scaling to 1. Missing values were imputed with the minimum.

Metabolomic profiling was performed on ovarian tumors obtained from obese and lean KpB mice treated with either vehicle or metformin. Samples were analyzed by Metabolon (Research Triangle Park, NC) according to their standard protocols 7 [92 (link)–95 (link)] and our previous work [96 (link)]. Briefly, unbiased global metabolomic profiling was achieved using methanol extracts of tumor tissues normalized to tissue weight. Analysis of extracts consisted of either ultrahigh performance liquid chromatography (Waters Corporation, Milford, MA) coupled with tandem mass spectrometry (UHPLC/MS/MS; Thermo-Finnigan, San Jose CA) in positive and negative ionization modes, or via gas chromatography/MS analysis (Thermo-Finnigan). Metabolites in tumor tissues were positively identified by matching chromatographic retention time, mass and MS/MS fragmentation patterns to a reference library of over 2500 purified, authenticated biochemicals. Data are presented as relative measures of “scaled intensity” and median scaling to 1. Missing values were imputed with the minimum.