A method to study column efficiency in indirect HPLC enantioseparation

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Abstract

Four ways are proposed for the comparison of true column performance for various C18 columns: plotting the resolution vs. the retention factor, plotting the resolution rate vs. retention factor, calculating the retention time needed for a given resolution, or the resolution obtained in a given time. The utilization of these methods is shown for eleven C18 columns in the separation of the enantiomers of erythro-β-methyl amino acids. The indirect chromatographic method involved pre-column derivatization with new chiral derivatizing agents (CDAs), such as (1S,2S)-1,3-diacetoxy-1-(4- nitrophenyl)-2-propyl isothiocyanate [(S,S)-DANI], and (S)-N-(4- nitrophenoxycarbonyl)phenylalanine methoxyethyl ester [(S)-NIFE], and the separation of the diastereomers formed on different types of C18 columns. The different columns were compared in systematic chromatographic examinations. The influence of various parameters of the columns on the resolution and the time of analysis are discussed.

Original languageEnglish
Pages (from-to)3227-3241
Number of pages15
JournalJournal of Liquid Chromatography and Related Technologies
Volume27
Issue number20
DOIs
Publication statusPublished - Dec 2 2004

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Keywords

  • (1S,2S)-1,3-Diacetoxy-1-(4-nitrophenyl)-2- propyl isothiocyanate
  • (S)-N-(4-Nitrophenoxycarbonyl)phenylalanine methoxyethyl ester
  • (S,S)-DANI
  • (S,S)-NIFE
  • Column efficiency
  • High performance liquid chromatography
  • erythro-β-Methyl amino acids

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Pharmaceutical Science
  • Clinical Biochemistry

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