1 / 16

Comprehensive Metabolite Profiling Using UHPLC-MS and GC-MS Techniques for Polar and Lipophilic Compounds

This study presents a detailed methodology for the identification and profiling of metabolites using advanced techniques such as GC-MS, UHPLC-MS, and NMR. The polar and lipophilic profiles of dry tissue samples were analyzed through targeted GC-MS deconvolution and metabolite identification. Techniques like AMDIS and spectral matching with custom libraries were employed for enhanced accuracy. Additionally, solid-phase extraction combined with UHPLC and NMR analysis were performed to elucidate complex metabolome profiles, ensuring robust identification of flavonoids and saponins, among other compounds.

jelani-sharp
Télécharger la présentation

Comprehensive Metabolite Profiling Using UHPLC-MS and GC-MS Techniques for Polar and Lipophilic Compounds

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  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

More Related