Adsorption behavior of a teicoplanin aglycone bonded stationary phase under harsh overload conditions

Matthias Bechtold, Attila Felinger, Martin Held, Sven Panke

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17 Citations (Scopus)


Silica-bonded teicoplanin aglycone allows enantioseparation of amino acids by reversed-phase liquid chromatography with a low organic solvent content. However, a reversible change in the adsorption behavior leading to a retention time shift (RTS) was observed when a preparative scale column was treated with harsh preparative chromatography-like conditions between finite-injection HPLC runs conducted under exactly the same conditions. This behavior was observed for all five investigated aliphatic and aromatic amino acids. In all cases, the retention times were prolonged after the overload conditions and the RTS was more pronounced for the later eluting d-enantiomer. We defined a standardized method for measuring the RTS and performed a systematic investigation on the influence of experimental conditions (type and concentration of pH modifier and organic modifier, temperature, pH) on the RTS. In this way a solvent composition - 90/10 50 mM NH4Ac pH 5.8/MeOH - was identified that yielded no observable shift in retention time after overload conditions for both enantiomers. In order to treat the observed phenomenon on a mechanistic level, we applied band profile analysis based on the stochastic theory of chromatography and identified two different enantioselective sites. When the band profile analysis was performed on elution profiles obtained from runs with prolonged retention time after harsh overload conditions, the retention time shift could be attributed to both differentiable types of adsorption sites. One site was found to make both, enantioselective and non-selective contributions.

Original languageEnglish
Pages (from-to)277-286
Number of pages10
JournalJournal of Chromatography A
Issue number1-2
Publication statusPublished - Jun 22 2007



  • Amino acid separation
  • Aqueous-organic phases
  • Chirobiotic TAG column
  • Influence of organic solvent on adsorption behavior
  • Macrocyclic glycopeptide stationary phases
  • Memory effect
  • Methionine separation

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

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