title: Determination of enantiomeric impurity in besifloxacin hydrochloride by chiral high-performance liquid chromatography with precolumn derivatization.
abstract: Besifloxacin hydrochloride is a novel chiral broad-spectrum fluoroquinolone developed for the treatment of bacterial conjunctivitis. R-besifloxacin hydrochloride is used in clinics as a consequence of its higher antibacterial activity. To establish an enantiomeric impurity determination method, some chiral stationary phases (CSPs) were screened. Besifloxacin enantiomers can be separated to a certain extent on Chiral CD-Ph (Shiseido Co., Ltd., Japan), Chiral AGP, and Crownpak CR (+) (Daicel Chemical IND., Ltd., Japan). However, the selectivity and sensitivity were both unsatisfactory on these three CSPs. Therefore, Chiral AGP, Chiral CD-Ph, and Crownpak CR (+) were not used in the enantiomeric impurity determination of besifloxacin hydrochloride. The separation of enantiomers of besifloxacin was further performed using a precolumn derivatization chiral high-performance liquid chromatography method. 2,3,4,6-Tetra-O-acetyl-beta-D-glucopyranosyl isothiocyanate was used as the derivatization reagent. Besifloxacin enantiomer derivates were well separated on a C(18) column (250 × 4.6 mm, 5 µm) with a mobile phase that consisted of methanol-KH(2)PO(4) buffer solution (20 mM; pH 3.0) (50:50, v/v). Selectivity, sensitivity, linearity, accuracy, precision, stability, and robustness of this method were all satisfied with the method validation requirement. The method was suitable for the quality control of enantiomeric impurity in besifloxacin hydrochloride.
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, Zhejiang, China.
authors.forename: Zhangting
authors.initials: Z
authors.lastname: Wang
authors.forename: Shengjia
authors.initials: S
authors.lastname: Wang
authors.forename: Feng
authors.initials: F
authors.lastname: Zhu
authors.forename: Zaixin
authors.initials: Z
authors.lastname: Chen
authors.forename: Lushan
authors.initials: L
authors.lastname: Yu
authors.forename: Su
authors.initials: S
authors.lastname: Zeng
mesh_terms: D001381:Azepines / Q000032:analysis* / Q000737:chemistry* / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D024841:Fluoroquinolones / Q000032:analysis* / Q000737:chemistry* / Q000302:isolation & purification; D017879:Isothiocyanates / Q000737:chemistry; D016014:Linear Models; D013237:Stereoisomerism
chemical_list: D001381:Azepines; D024841:Fluoroquinolones; D017879:Isothiocyanates; C041943:2,3,4,6-tetra-O-acetylglucopyranosylisothiocyanate; C522124:besifloxacin