Multianalyte determination of the kinetic rate constants of drug–cyclodextrin supermolecules by high performance affinity chromatography

• Composition of the mobile phase greatly affected the determination of kd,app.
• The multianalyte approach was employed to measure kd,app values.
• The multianalyte approach was statistically equivalent with that of the single compound analysis.
• The approach can be employed for the efficient determination of kd,app with less variance caused by CD column bleeding.

The kinetics of the dissociation is fundamental to the formation and the in vivo performance of cyclodextrin supramolecules. The individual determination of the apparent dissociation rate constant (kd,app) using high performance affinity chromatography (HPAC) is a tedious process requiring numerous separate studies and massive data fitting. In this study, the multianalyte approach was employed to simultaneously measure the kd,app values of three drugs through one injection based on the investigation of the dependence of drug–cyclodextrin interaction kinetics on the mobile phase composition. As a result, the kd,app values increased when decreasing the ion strength, increasing the ionization of drugs and adding extra organic solvents. The values of kd,app for acetaminophen, phenacetin and S-flurbiprofen estimated by the multianalyte approach were 8.54 ± 1.81, 5.36 ± 0.94 and 0.17 ± 0.02 s−1, respectively, which were in good agreement with those determined separately (8.31 ± 0.58, 5.01 ± 0.42 and 0.15 ± 0.01 s−1). For both of the single and multiple flow rate peak profiling methods, the results of the multianalyte approach were statistically equivalent with that of the single compound analysis for all of the three drugs (p > 0.05). The multianalyte approach can be employed for the efficient evaluation of the drug–cyclodextrin kinetics with less variance caused by cyclodextrin column bleeding.