Supercritical fluid extraction of amino acids from birch (Betula pendula Roth) leaves and their liquid chromatographic determination with fluorimetric detection

J Sep Sci. 2008 May;31(8):1363-73. doi: 10.1002/jssc.200700560.

Abstract

A method for supercritical fluid extraction (SFE) of amino acids was adapted and optimal experimental conditions were selected for a matrix consisting of dry leaves. The matrix-dependent SFE method uses a mixture of MeOH-H(2)O-acetonitrile (10:10:1 v/v/v) as a modifier (0.5 mL in situ, 300 muL on-line) at 70 degrees C and 40 MPa and no HCl is needed as an entrainer. The amino acids were quantified using high-performance liquid chromatography with fluorimetric detection (HPLC/FLD) after gradient elution on Zorbax Eclipse AAA columns (4.6x150 mm, 3.5 mum) with aqueous Na(2)HPO(4 )buffer of pH 7.8 and ACN-MeOH-water as a mobile phase. In comparison with Soxhlet extraction, SFE gave higher recovery and selectivity, but it required longer extraction time (90 min) and it was more labor-intensive (clean-up step after the pre-concentration). Both methods should be used separately or in combination according to the matrix, number of samples, and levels of ballast compounds.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetonitriles / analysis*
  • Amino Acids / analysis*
  • Amino Acids / isolation & purification
  • Betula / metabolism*
  • Chromatography, Liquid / methods*
  • Chromatography, Supercritical Fluid / methods*
  • Czech Republic
  • Hydrogen-Ion Concentration
  • Methanol / chemistry
  • Plant Leaves / metabolism
  • Pressure
  • Reproducibility of Results
  • Solvents / chemistry
  • Temperature
  • Time Factors
  • Water / chemistry

Substances

  • Acetonitriles
  • Amino Acids
  • Solvents
  • Water
  • Methanol
  • acetonitrile