Strategy for automated analysis of dynamic metabolic mixtures by NMR. Application to an insect venom

Anal Chem. 2007 Oct 15;79(20):7748-52. doi: 10.1021/ac0711586. Epub 2007 Sep 7.

Abstract

Elucidation of the composition of chemical-biological samples is a main focus of systems biology and metabolomics. Due to the inherent complexity of these mixtures, reliable, efficient, and potentially automatable methods are needed to identify the underlying metabolites and natural products. Because of its rich chemical information content, nuclear magnetic resonance (NMR) spectroscopy has a unique potential for this task. Here we present a generalization and application of a recently introduced NMR data collection, processing, and analysis strategy that circumvents the need for extensive purification and hyphenation prior to analysis. It uses covariance TOCSY NMR spectra measured on a 1-mm high-temperature cryogenic probe that are analyzed by a spectral trace clustering algorithm yielding 1D NMR spectra of the individual components for their unambiguous identification. The method is demonstrated on a metabolic model mixture and is then applied to the unpurified venom mixture of an individual walking stick insect that contains several slowly interconverting and closely related metabolites.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Automation
  • Insecta / chemistry
  • Insecta / metabolism*
  • Magnetic Resonance Spectroscopy / methods*
  • Molecular Structure
  • Venoms / analysis*
  • Venoms / chemistry

Substances

  • Venoms