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Advanced Metabolite Profiling via GC-MS and UHPLC-MS Techniques in Dry Tissue Analysis

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This study explores the comprehensive analysis of polar and lipophilic metabolites in dry tissue samples through advanced methods including GC-MS and UHPLC-MS. Derivatization is performed using different solvent systems, followed by metabolite deconvolution and identification via AMDIS and spectral matching with both commercial and custom libraries. Techniques like solid-phase extraction and NMR are integrated for enhanced metabolomics profiling, achieving detailed coverage of biomolecules such as flavones and saponins, contributing valuable insights into biochemical pathways.

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Advanced Metabolite Profiling via GC-MS and UHPLC-MS Techniques in Dry Tissue Analysis

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  1. 10 mg Dry Tissue MeOH +H2O & CHCl3 H2O/MeOH/CHCl3 H2O/MeOH CHCl3 Polar Lipophilic GCMS derivatization

  2. POLAR Profile LIPOPHILIC Profile GCMS deconvolution and metabolite identification are performed using AMDIS and spectral matching with commercial & custom EI GC-MS Libraries UHPLC-MS deconvolution using Bruker Dissect, Waters MarkerLynx, and Agilent Mass Hunter. Metabolite identification via spectral matching with Custom (-)ESI UHPLC-MS & MS/MS Libraries of Authentic Compounds.

  3. UHPLC-UV-MS-SPE-NMR Bruker/Spark Holland Prospeckt 2 Solid Phase Extraction Mass Spectrometer Make-up Pump 1.5 ml/min H20 UHPLC Analytical Column 0.560 ml/min UV waste 600 MHz NMR With Cryoprobe

  4. UHPLC-UV-MS-SPE-NMR Bruker/Spark Holland Prospeckt 2 Solid Phase Extraction Make-up Pump 1.5 ml/min H20 UHPLC Mass Spectrometer Analytical Column 0.560 ml/min UV waste 600 MHz NMR With Cryoprobe

  5. UHPLC-MS-SPE-NMR Bruker Maxis Qtof-MS/MS MS & SPE Splitter Gilson liquid handler For SPE eluant collection Dr. Dennis Fine Waters AcquityIClass UHPLC With photodiode array detector Prospect 2 Solid Phase Extraction Unit w/ 2 x 96 cartridges & deuterated solvent elution

  6. LC/QTOF SPE Profile of Flavone/Saponin Mix

  7. LC/QTOF SPE Profile of Flavone/Saponin Mix Reference Spectrum of authentic umbelliferone ~900 ngUmbelliferone1H NMR

  8. Current M. truncatula Metabolome Coverage Profiled Level#1 IDS* Shoot GCMS Polar 300 136 Shoot GC-MS nonpolar 250 112 GC-MS Shoot waxes 200 48 Volatile SPME-GCMS 150 74 UHPLC-QTofMS 200 75 Hormones GCMS/LC-QqQMS 25 25 = 1125 Total peaks profiled = 470 Total Peaks Profiled *IDs based upon co-characterization with authentic standards and per Sumner et al, 2007. Proposed minimum reporting standards for chemical analysis, Metabolomics, 3(3) 211-221

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