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.

Chiral separation of quinolones by liquid chromatography and capillary electrophoresis

“…Asymmetric peaks of besifloxacin enantiomers on a Chiral‐AGP column were observed by Wang et al. who also used sodium dihydrophosphate buffer solution modified with 3% of an organic solvent, acetonitrile.…”

“…So, Wang et al. while developing a method to determine enantiomeric impurity in ( R )‐besifloxacin screened a number of polysaccharide, protein, and crown ether‐based CSPs but could not find appropriate conditions for acceptable resolution of the R and S enantiomers. Instead, they achieved good separation after the derivatization of the sample with 2,3,4,6‐tetra‐ O ‐acetyl‐β‐ d ‐glucopyranosyl isothiocyanate following chromatography on a C 18 column.…”

“…Wang et al. had unsuccessfully tested nine polysaccharide‐based CSPs under the normal phase conditions and six polysaccharide‐based CSPs under RP conditions in the separation of besifloxacine enantiomers. Yeole et al.…”

Uv-HPLC Method Development and Validation for Quantification of Besifloxacin Hydrochloride

“…The purpose of this proposed work was to develop and validate the method using ultraviolet-high performance liquid chromatographic (UV-HPLC) for quantification of besifloxacin hydrochloride. In literature, other analytical methods described for the quantitative analysis of besifloxacin hydrochloride but no report found using UV-HPLC method [3][4][5][6][7][8][9]. The newly developed method was a unique advantage over the other available methods, as it is simple, accurate, precise, and specific for quantitative determination of besifloxacin hydrochloride in pharmaceutical dosage form.…”

Forming of Diastereoisomer With L-Isoleucin to Separate Ofloxacin Enantiomer Using Preparative Liquid Chromatography

“…Enantiomers are two types of chemical species with identical molecule but different from each other as mirror images that do not coincide (Wang et al, 2012). Enantiomer of drugs have toxicological and pharmacological effects that are different so that in most cases the pharmacological effects restricted to only one enantiomer (eutomer) while the unwanted enantiomer (distomer) have side effects or even toxic (Schmid & Gübitz, 2011)..…”