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Observed retention factors were determined for leukotriene B 4 , leukotriene E 4 and N-acetylleukotriene E 4 using mobile phases with different percentages of organic modifier in the pH range from 2 to 12. A two-stage least squares fitting procedure for fitting a general equation to the observed retention factors was used to calculate the dissociation constants at each percentage...
A general equation relating the observed retention factor to the pH of the mobile phase, the dissociation constants, and the retention factors of the different ionic species has been derived. This equation is applicable to polyprotic weak acid and base dissociation events, that is, the secondary equilibria existing in the HPLC mobile phase. It is written as: where the k r values are the...
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