Theoretical opportunities and actual limitations of pH gradient HPLC

Kaliszan, Roman; Wiczling, Pawel
June 2005
Analytical & Bioanalytical Chemistry;Jun2005, Vol. 382 Issue 3, p718
Academic Journal
In a series of reports published recently by our laboratory comprehensive theory and experimental conditions were established for reversed-phase high-performance liquid chromatography (RP HPLC) employing the programmed pH gradient of mobile phase. A procedure was developed providing, rapidly and conveniently, the acidity (pKa) of weak acids and bases and their lipophilicity (hydrophobicity) logkw. The basis of the double-gradient RP HPLC, employing simultaneous gradients of organic modifier content and mobile phase pH, was also elaborated. The fundamentals of the approach are presented briefly and systematically and its advantages and limitations are discussed. It is demonstrated that the newly introduced pH gradient method increases the analytical versatility of RP HPLC and our understanding of its physicochemical basis.


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