A novel method of sample homogenization with the use of a microtome-cryostat apparatus

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Abstract

Quantitative metabolomics places high demands on sample preparation, including a high degree of metabolite extraction and controlled sample weight. In respect to elastic collagen-rich tissues, the existing methods of sample homogenization poorly fit these demands due to incomplete homogenization, sample material loss, or metabolite degradation. Herein, a novel method based on the use of a microtome-cryostat apparatus is proposed. The performance of the cryotome method is compared with the results obtained with the use of a vortex bead beating. NMR-based metabolomic analysis shows that the extraction efficiency and the data scattering for both methods of sample preparation are similar. However, the heat generation during the bead beating causes the destruction of thermally-unstable compounds; besides, it may cause protein hydrolysis, leading to an artificial increase in the amino acid level. The cryotome method of sample homogenization does not cause sample heating, and it seems to be ideal for elastic tissues.

Original languageEnglish
Pages (from-to)37809-37817
Number of pages9
JournalRSC Advances
Volume9
Issue number65
DOIs
Publication statusPublished - 21 Nov 2019

Keywords

  • NUCLEAR-MAGNETIC-RESONANCE
  • EXTRACTION STRATEGIES
  • METABOLITE EXTRACTION
  • METABOLOMICS
  • NMR
  • SERUM
  • QUANTIFICATION
  • DEGRADATION
  • COENZYMES

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