Enantioseparation and ecotoxicity evaluation of ibrutinib by Electrokinetic Chromatography using single and dual systems

In this work, two chiral methods enabling the separation of ibrutinib enantiomers were developed by Electrokinetic Chromatography. A cyclodextrin (CD) or a mixture of the CD and a chiral ionic liquid (CIL) was used as chiral selector. Using the single CD system, seven neutral and six anionic CDs were tested in a formate buffer at pH 3.0 working in positive and negative polarity, respectively. The use of sulfated-γ-CD (S-γ-CD) and negative polarity originated the best results considering analysis time and enantioresolution. The optimization of the experimental conditions allowed obtaining the separation of ibrutinib enantiomers in an analysis time of 4.2 min with an enantioresolution value of 1.5. The effect of the addition of fifteen CILs on the enantioresolution was evaluated showing that both analysis time and enantioresolution were generally increased. A mixture of S-γ-CD and [TMA][L-Lys] was selected which provided the separation of ibrutinib enantiomers in 8.1 min with an enantioresolution value of 3.3 under the same experimental conditions as in the case of using the single CD system. The enantiomeric impurity (S-ibrutinib) was the first-migrating isomer when using the single CD and the combined CD/CIL systems, as corresponds to the most desirable situation. Both chiral methods allowed the detection of the enantiomeric impurity up to a 0.1% as established by the International Council on Harmonization. After establishing the analytical characteristics of both chiral methodologies developed, they were applied to the enantiomeric determination of ibrutinib in a pharmaceutical formulation for hospital use marketed as pure enantiomer (R-ibrutinib) and to evaluate the stability and ecotoxicity of racemic ibrutinib and R-ibrutinib on Daphnia magna. The developed methodologies enabled, for the first time, the rapid chiral quantitation of ibrutinib in abiotic and biotic matrices.

Chiral separation; Cyclodextrin-electrokinetic chromatography; Ecotoxicity; Ibrutinib; Ionic liquid; Pharmaceutical formulation.