title: Development and Validation of a Normal Phase Chiral HPLC Method for Analysis of Afoxolaner Using a Chiralpak® AD-3 Column.
abstract: 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®AD-3 column (150 × 4.6 mm I.D.) maintained at 35°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. ® All marks are the property of their respective owners.
authors.affiliation: Merial Inc., A Sanofi Company, 631 Rt 1 South, North Brunswick, NJ 08902, USA jinyou.zhuang@merial.com.
authors.forename: Jinyou
authors.initials: J
authors.lastname: Zhuang
authors.affiliation: Merial Inc., A Sanofi Company, 631 Rt 1 South, North Brunswick, NJ 08902, USA.
authors.forename: Satish
authors.initials: S
authors.lastname: Kumar
authors.affiliation: Merial Inc., A Sanofi Company, 631 Rt 1 South, North Brunswick, NJ 08902, USA.
authors.forename: Abu
authors.initials: A
authors.lastname: Rustum
mesh_terms: D000688:Amylose / Q000031:analogs & derivatives* / Q000737:chemistry; D002623:Chemistry Techniques, Analytical / Q000379:methods* / Q000592:standards; D002851:Chromatography, High Pressure Liquid; D007555:Isoxazoles / Q000032:analysis* / Q000302:isolation & purification; D009281:Naphthalenes / Q000032:analysis* / Q000302:isolation & purification; D048448:Phenylcarbamates / Q000737:chemistry*; D015203:Reproducibility of Results; D013237:Stereoisomerism
chemical_list: D007555:Isoxazoles; D009281:Naphthalenes; D048448:Phenylcarbamates; C000589002:afoxolaner; C080033:Chiralpak AD; D000688:Amylose