Development and Validation of a Normal Phase Chiral HPLC Method for Analysis of Afoxolaner Using a Chiralpak (R) AD-3 Column

摘要

Afoxolaner is a new antiparasitic molecule from the isoxazoline family that acts on the insect acarine gamma-aminobutyric acid and glutamate receptors. Isoxazoline family of compounds has been employed as active pharmaceutical ingredient in drug products prescribed for control of fleas and ticks in dogs. Afoxolaner with a chiral center at isoxazoline ring exists as a racemic mixture. A normal phase chiral high performance liquid chromatography analytical method has been developed to verify that afoxolaner is a racemic mixture as demonstrated by specific rotation, as well as to determine enantiomeric purity of single enantiomer samples. A Chiralpak (R) AD-3 column (150 x 4.6mm I.D.) maintained at 35 degrees C was used in the method. Analytes were analyzed with an isocratic elution using n-Hexane/IPA/MeOH (89:10:1, v/v/v) as the mobile phase with a detection wavelength of 312 nm. Desired separation of the two enantiomers was achieved in < 10 minutes with resolution and selectivity factors of 5.0 and 1.54, respectively. The analytical method was appropriately validated according to ICH guidelines for its intended use. (R) All marks are the property of their respective owners.