•The HPAC, SPR and SPRi methods were compared from drug-CD interaction measurements.•A large deviation in data from 3 methods was observed for 18 insoluble drugs.•The performance and importance of each method was thoroughly discussed.•The article will help to select the appropriate methodology for future measurements.Three advanced methods, high performance affinity chromatography (HPAC), surface plasmon resonance (SPR) and surface plasmon resonance imaging (SPRi) were compared and evaluated for determining the drug-cyclodextrin (CD) interactions herein. In total, 18 sparingly soluble drugs were selected for this comparative study. The three methods share a unique connection in the working principles and strategies. The same strategies of CD fixation onto solid phase were used in HPAC and SPR for the measurements, whereas, the SPR and SPRi share identical working principles. However, whilst these relationships are evident, no strong correlation was found between kinetic constants obtained from the three methods: Four drugs, namely, prednisolone, pseudolaric acid B, diazepam and gramisetron failed to show any response on SPR, whereas, the kinetics parameters from SPRi and HPAC were successfully measured. From a comparative review of all the kinetic data, random results without any trends were observed (ka, kd and KA) regardless of the relationships between the three methods: It is apparent that the measurement conditions (volume, flow rate, buffers), non-specific adsorption and experimental procedures had a strong impact on the generated data. The relative advantages and limitations of each method are critically presented on the basis of generated data. This comparative study provides a basis to further upgrade these techniques for confident measurement of drug-CDs interactions.

•Structured γ-CD-MOFs as stationary phase was used for drug loading and retention.•High correlation between retention and drug loading was observed for γ-CD-MOFs.•New method is highly efficient to the conventional ways of drug loading into MOFs.Drug loading into γ-cyclodextrin-metal organic frameworks (γ-CD-MOFs) using the impregnation approach is a laborious process. In this study, a γ-CD-MOF construct (2–5 μm particle diameter) was used as the stationary phase under HPLC conditions with the aim to correlate retention properties and drug loading capability of the CD-based structure. Ketoprofen, fenbufen and diazepam were chosen as model drugs with m-xylene as a control analyte to investigate the correlation of drug loading and their chromatographic behaviour in the γ-CD-MOF column. Furthermore, γ-CD itself was also prepared as the stationary phase by coupling with silica in the column to illustrate the enhanced interaction between drugs and γ-CD-MOF as a reference. The retention and loading efficiency of the drugs were determined with different ratios of hexane and ethanol (10:90, 20:80, 50:50, 80:20, 90:10, v/v) at temperatures of 20, 25, 30 and 37 °C. With the increment in hexane content, the loading efficiency of ketoprofen and fenbufen increased from 2.39 ± 0.06% to 4.38 ± 0.04% and from 5.82 ± 0.94% to 6.37 ± 0.29%, respectively. The retention time and loading efficiency of ketoprofen and diazepam were the lowest at 30 °C while those of fenbufen had the different tendency. The excellent relation between the retention and loading efficiency onto γ-CD-MOF could be clearly observed through mobile phase and temperature investigation. In conclusion, a highly efficient chromatographic method has been established to evaluate the drug loading capability of γ-CD-MOF.

•A HPAC-MS/MS method was developed to determine kd,app of drug-cyclodextrin interaction with twenty drugs in one injection.•The HPAC-MS/MS method was employed to determine the kd,app of twenty drugs in one injection with high-throughput yield.•Good correlationship was found between kd,app measured by single compound analysis method and high-throughput HPAC-MS/MS approach.•Drug-CD interaction kinetics under different conditions was also investigated based on the comprehensive quantification of ka,app, kd,app and Ka.The individual determination of the apparent dissociation rate constant (kd,app) using high performance affinity chromatography (HPAC) is a tedious process requiring numerous separate tests and massive data fitting, unable to provide the apparent association rate constant (ka) and equilibrium binding constant (Ka). In this study, a HPAC with mass spectrometry detection (HPAC-MS/MS) was employed to determine the drug-cyclodextrin (CD) interaction kinetics with low sample loading quantity (<10 ng per injection for single compound) and high-throughput yield as twenty drugs determined in one injection. The kd,app measured by HPAC-MS/MS approach were 0.89 ± 0.07, 4.34 ± 0.01, 1.48 ± 0.01 and 7.77 ± 0.04 s−1 for ketoprofen, trimethoprim, indapamide and acetaminophen, with kd,app for acetaminophen consistent with that from the HPAC method with UV detector in our previous studies. For twenty drugs with diverse structures and chemical properties, good correlationship was found between kd,app measured by single compound analysis method and high-throughput HPAC-MS/MS approach, with the correlation coefficient of 0.987 and the significance F less than 0.001. Comprehensive quantification of ka,app, kd,app and Ka values was further performed based on the measurement of kd,app by peak profiling method and Ka by the peak fitting method. And the investigation of the drug-CD interaction kinetics under different conditions indicated that the column temperature and mobile phase composition significantly affected the determination of ka,app, kd,app and Ka while also dependent on the acidity and basicity of drugs. In summary, the high-throughput HPAC-MS/MS approach has been demonstrated high efficiency in determination of the drug-CD primary interaction kinetic parameter, especially, kd,app, being proven as a novel tool in screening the right CD for the solubilization of the right drug.

•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.

•A modified HPAC method is proposed to determine kd of cyclodextrin supramolecules.•The plate height for the theoretical nonretained substance is estimated.•The kd for acetaminophen and sertraline are determined as 1.78 and 1.91 s−1.•The kd value is consistent with that determined by capillary electrophoresis.It is challenging and extremely difficult to measure the kinetics of supramolecular systems with extensive, weak binding (Ka < 105 M−1), and fast dissociation, such as those composed of cyclodextrins and drugs. In this study, a modified peak profiling method based on high performance affinity chromatography (HPAC) was established to determine the dissociation rate constant of cyclodextrin supramolecular systems. The interactions of β-cyclodextrin with acetaminophen and sertraline were used to exemplify the method. The retention times, variances and the plate heights of the peaks for acetaminophen or sertraline, conventional non-retained substance (H2O) on the β-cyclodextrin bonded column and a control column were determined at four flow rates under linear elution conditions. Then, plate heights for the theoretical non-retained substance were estimated by the modified HPAC method, in consideration of the diffusion and stagnant mobile phase mass transfer. As a result, apparent dissociation rate constants of 1.82 (±0.01) s−1 and 3.55 (±0.37) s−1 were estimated for acetaminophen and sertraline respectively at pH 6.8 and 25 °C with multiple flow rates. Following subtraction of the non-specific binding with the support, dissociation rate constants were estimated as 1.78 (±0.00) and 1.91 (±0.02) s−1 for acetaminophen and sertraline, respectively. These results for acetaminophen and sertraline were in good agreement with the magnitude of the rate constants for other drugs determined by capillary electrophoresis reported in the literature and the peak fitting method we performed. The method described in this work is thought to be suitable for other supramolecules, with relatively weak, fast and extensive interactions.