Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1048, Issue 2, Pages 183-193Publisher
ELSEVIER
DOI: 10.1016/j.chroma.2004.07.022
Keywords
mobile phase composition; method development; ionizable compounds; retention behaviour
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Optimizing separation of ionizable compounds in order to find robust conditions has become an important part of method development in liquid chromatography. This work is an attempt to explain the observed variations of retention of acid and basic compounds with the organic modifier content in the mobile phase, according to various factors: the type of modifier, the type of buffer, the temperature and of course the type of solute. This is done by considering the variation of the so-called chromatographic pK(a) which refers to the pH measured in the aqueous medium and is determined from retention data. A procedure is described that accurately relates, from nine experiments, retention to solvent composition and pH. The limits of such a procedure are evaluated and two examples of optimized separations of basic compounds are given. (C) 2004 Elsevier B.V. All rights reserved.
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