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Anion-Exchange Chromatography: History
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Contributor: Helena Kang

Anion-exchange chromatography is a liquid-chromatographic separation in which negatively charged analytes compete for positively charged sites on a stationary phase and are eluted by a competing anion in the mobile phase [1][2]. In the chromatography of natural products and related ionic species the concept is bounded by that electrostatic retention mechanism: the stationary phase is an anion exchanger (typically a quaternary ammonium or related cationic group), and the solutes that are retained are anions. Essential features are a charged surface whose ion-exchange capacity sets the load, a buffered eluent whose pH controls the charge state of weakly acidic analytes, and a salt or pH gradient that displaces bound anions in order of their effective affinity [2][3]. High-performance anion-exchange chromatography of carbohydrates, often paired with pulsed amperometric detection, exploits the weak acidity of sugar hydroxyl groups at high pH to retain otherwise neutral monosaccharides and polysaccharides as oxyanions [2][3][4]. The technique is distinguished from cation-exchange chromatography by the sign of the exchanged ion, from reversed-phase chromatography by the electrostatic rather than hydrophobic retention mechanism, and from ion-exclusion chromatography by the direction of the electrostatic interaction. Conceptual limits exclude electrophoretic separations that do not employ a packed or immobilized ion-exchange bed.

  • anion-exchange chromatography
  • ion exchanger
  • carbohydrate analysis
  • electrostatic retention

Chromatography in Natural Products • Analytical Chemistry • Chemistry • Physical Sciences

References

  1. Titus H.J. Huisman; Andrée M. Dozy; Studies on the heterogeneity of hemoglobin. J. Chromatogr. A 1965, 19, 160-169, 10.1016/s0021-9673(01)99434-8.
  2. Mark R. Hardy; R.Reid Townsend; Yuan C. Lee; Monosaccharide analysis of glycoconjugates by anion exchange chromatography with pulsed amperometric detection. Anal. Biochem. 1988, 170, 54-62, 10.1016/0003-2697(88)90089-9.
  3. Y.C. Lee; High-performance anion-exchange chromatography for carbohydrate analysis. Anal. Biochem. 1990, 189, 151-162, 10.1016/0003-2697(90)90099-u.
  4. Gerhard A. De Ruiter; Henk A. Schols; Alfons G.J. Voragen; Frank M. Rombouts; Carbohydrate analysis of water-soluble uronic acid-containing polysaccharides with high-performance anion-exchange chromatography using methanolysis combined with TFA hydrolysis is superior to four other methods. Anal. Biochem. 1992, 207, 176-185, 10.1016/0003-2697(92)90520-h.
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