The selection of suitable columns for a reversed-phase liquid chromatographic separation of beta-lactam antibiotics and related substances via chromatographic column parameters

J Chromatogr A. 2014 Jan 3:1323:87-96. doi: 10.1016/j.chroma.2013.11.005. Epub 2013 Nov 11.

Abstract

The selection of RP-LC columns suitable for a particular analysis in official compendia is difficult as only a general description of the stationary phase in the description of a LC method is given. General methods to characterize RP-LC columns often assume that each of the column parameters is equally important. This can cause the user to select columns inappropriate for particular analyses. This paper focuses on the relationship between the critical peak pairs and the column parameters (H, S, A, B, and C) in the Snyder/Dolan column characterization methodology to find the key parameters influencing real separations. Some varieties of β-lactam antibiotics and their related compounds were used as test compounds. We found column parameter A to be the most important factor affecting their separation. Parameters B and C also played an important role in some separation processes. This indicated that the hydrogen bonding of column and solute can directly affect the separation of β-lactam antibiotics. Choosing columns for which column parameter A is near 0.1 can facilitate the ideal separations of impurities from β-lactam antibiotics. The most suitable column for any common pharmaceutical analysis could be selected easily if the key column parameters would be given in the description of the chromatographic method. For these reasons, key column parameters should be listed in the monographs of official compendia.

Keywords: Column parameter; Critical peak pair; Selection of chromatographic column; β-Lactam antibiotic.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Anti-Bacterial Agents / chemistry
  • Anti-Bacterial Agents / isolation & purification*
  • Cefotaxime / chemistry
  • Cefotaxime / isolation & purification
  • Chromatography, Reverse-Phase / instrumentation*
  • Chromatography, Reverse-Phase / methods*
  • Hydrogen Bonding
  • beta-Lactams / chemistry
  • beta-Lactams / isolation & purification*

Substances

  • Anti-Bacterial Agents
  • beta-Lactams
  • Cefotaxime