title: Simple Isocratic HPLC Method for Determination of Enantiomeric Impurity in Besifloxacin Hydrochloride.
abstract: Besifloxacin is a unique chiral broad-spectrum flouroquinolone used in the treatment of bacterial conjunctivitis. R-form of besifloxacin hydrochloride shows higher antibacterial activity as compared to the S-isomer. Therefore, it is necessary to establish chiral purity. To establish chiral purity a high-performance liquid chromatography (HPLC) method for determination of R-besifloxacin and S-besifloxacin (BES impurity A) was developed and validated for in-process quality control and stability studies. The analytical performance parameters such as linearity, precision, accuracy, specificity, limit of detection (LOD), and lower limit of quantification (LOQ) were determined according to International Council for Harmonization ICH Q2(R1) guidelines. HPLC separation was achieved on Chiralpak AD-H (250 x 4.6 mm, 5 μm) column using n-heptane: ethanol: ethylenediamine: acetic acid (800:200:0.5:0.5) (v/v/v/v) as the mobile phase in an isocratic elution. The eluents were monitored by UV/Visible detector at 290 nm. The resolution between S-isomer and besifloxacin hydrochloride was more than 2.0. Based on a signal-to-noise ratio of 3 and 10 the LOD of besifloxacin was 0.30 μg/mL, while the LOQ was 0.90 μg/mL. The calibration curves were linear in the range of 0.9-7.5 μg/mL. Precision of the method was established within the acceptable range. The method was suitable for the quality control enantiomeric impurity in besifloxacin hydrochloride. Chirality 28:628-632, 2016. © 2016 Wiley Periodicals, Inc.
authors: Analytical Research Department, Micro Labs, Bommasandra, Bangalore.
authors: G Pradeep
authors: 0000-0001-9948-3150
authors: GP
authors: Kumar
authors: Analytical Research Department, Micro Labs, Bommasandra, Bangalore.
authors: Vishal
authors: V
authors: Srivastava
authors: Analytical Research Department, Micro Labs, Bommasandra, Bangalore.
authors: Kiran
authors: K
authors: Khandelwal
authors: Analytical Research Department, Micro Labs, Bommasandra, Bangalore.
authors: Rajesh
authors: R
authors: Kumar
authors: Analytical Research Department, Micro Labs, Bommasandra, Bangalore.
authors: S G
authors: SG
authors: Hiriyanna
authors: Analytical Research Department, Micro Labs, Bommasandra, Bangalore.
authors: Ajay
authors: A
authors: Kumar
authors: API R&D Centre, Micro Labs, Bommasandra, Bangalore.
authors: Pramod
authors: P
authors: Kumar
mesh_terms: D000688:Amylose / Q000031:analogs & derivatives; D001381:Azepines / Q000032:analysis* / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D024841:Fluoroquinolones / Q000032:analysis* / Q000737:chemistry*; D057230:Limit of Detection; D048448:Phenylcarbamates; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013237:Stereoisomerism; D014466:Ultraviolet Rays
chemical_list: D001381:Azepines; D024841:Fluoroquinolones; D048448:Phenylcarbamates; C080033:Chiralpak AD; D000688:Amylose; C522124:besifloxacin
keywords: besifloxacin hydrochloride; chiral; conjunctivitis; enantiomeric purity