Validation of the accuracy of the perturbation peak method for determination of multicomponent adsorption isotherm parameters in LC.

The isotherm parameters were for the first time determined for a quaternary mixture. This was done by the perturbation peak (PP) method using racemic mixtures of methyl and ethyl mandelate enantiomers. One complication with the PP method is that the traditional blank injection technique makes all perturbation peaks, except one, vanish at moderately nonlinear concentration plateaus. Therefore, we devised a new injection technique that made all four peaks on a quaternary component concentration plateau detectable, thereby making the determination of multicomponent competitive isotherm parameters possible. The measured quaternary perturbation data fitted well to the bi-Langmuir isotherm model and excellent agreement was found between experimental and simulated single-component and multicomponent profiles, thus validating the method and the determined isotherm parameters. The method (i) is valuable for computer-assisted optimization of preparative chiral chromatography and (ii) opens the possibility of quantifying competitive drug-target interactions for chiral drugs directly on racemic mixtures, which today is impossible with any nonlabeled technique including surface plasmon resonance technology.