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Ion-pairing high-performance liquid chromatographic method for the detection of N-acetylaspartate and N-acetylglutamate in cerebral tissue extracts.

Abstract
An ion-pairing high-performance liquid chromatographic method for the determination of N-acetylaspartate and N-acetylglutamate using a C-18 column and a UV detection at 210 nm wavelength, by means of a diode array detector, is presented. A buffer containing 2.8 mM tetrabutylammonium hydroxide, 25 mM KH(2)PO(4), 1.25% methanol, pH 7. 00, is utilized for the isocratic separation of these N-acetylated amino acids, at a flow rate of 1 ml/min and a column temperature of 23 degrees C. The suitability of this chromatographic separation (without additional chromatographic steps prior to HPLC assay) to monitor variations both of N-acetylaspartate and of N-acetylglutamate in perchloric acid brain extracts from rats subjected to the impact acceleration model of diffuse brain injury is also reported. According to the data presented, this HPLC method allows the separation of the two N-acetylated amino acids considered from the many possible interfering compounds, commonly present in extracts of cerebral tissue, which have high extinction coefficients at 210 nm wavelength. Values of N-acetylaspartate and N-acetylglutamate determined by this method showed that cerebral trauma negatively affects both compounds, according to the severity of trauma itself.
AuthorsB Tavazzi, R Vagnozzi, D Di Pierro, A M Amorini, G Fazzina, S Signoretti, A Marmarou, I Caruso, G Lazzarino
JournalAnalytical biochemistry (Anal Biochem) Vol. 277 Issue 1 Pg. 104-8 (Jan 01 2000) ISSN: 0003-2697 [Print] United States
PMID10610694 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
CopyrightCopyright 2000 Academic Press.
Chemical References
  • Glutamates
  • Tissue Extracts
  • Aspartic Acid
  • N-acetylaspartate
  • N-acetylglutamic acid
Topics
  • Animals
  • Aspartic Acid (analogs & derivatives, analysis, metabolism)
  • Brain Chemistry
  • Brain Injuries (metabolism)
  • Chromatography, High Pressure Liquid (methods)
  • Glutamates (analysis, metabolism)
  • Male
  • Rats
  • Rats, Wistar
  • Tissue Extracts (chemistry)

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