title: Validated chiral high-performance liquid chromatography method for a novel anti-methicillin-resistant Staphylococcus aureus fluoroquinolone WCK 771.
abstract: A sensitive, simple, specific, precise, accurate and rugged method for the assay and determination of enantiomeric purity of S-(-)-9-fluoro-6,7-dihydro-8-(4-hydroxypiperidin-1-yl)-5-methyl-1-oxo-1H,5H-benzo[i,j]quinolizine-2-carboxylic acid L-arginine salt tetrahydrate (WCK 771) in bulk drug has been developed. The method is RP-HPLC using endcapped C-18 stationary phase and chiral mobile phase. Chirality to the mobile phase was imparted with addition of beta-cyclodextrin. The UV-vis detector was operated at 290 nm. The flow rate of mobile phase was 2 ml/min. The method offers excellent separation of two enantiomers with resolution more than 2 and tailing factor less than 1.5. The method was validated for the assay of WCK 771 and quantification of R-(+)-enantiomer impurity in bulk drug. The calibration curves showed excellent linearity over the concentration range of 0.05-0.15 mg/ml for WCK 771 and 0.5-7.5 microg/ml for R-(+)-enantiomer. The precision (RSD) of the assay was 0.23%. The limit of detection and limit of quantitation of the method for WCK 771 were 0.015 and 0.06 microg/ml, respectively. The limit of detection and limit of quantitation for R-(+)-enantiomer were 0.025 and 0.09 microg/ml, respectively. The average recovery of the R-(+)-enantiomer was 100.5%. Same method was applied for the assay and determination of enantiomeric purity of WCK 771 in the intravenous formulation.
authors.affiliation: Wockhardt Limited, Wockhardt Research Center, Analytical, Drug Discovery, D-4, MIDC Area, Chikalthana, Aurangabad, Maharashtra 431210, India. ryeole@wockhardtin.com
authors.forename: R D
authors.initials: RD
authors.lastname: Yeole
authors.forename: A S
authors.initials: AS
authors.lastname: Jadhav
authors.forename: K R
authors.initials: KR
authors.lastname: Patil
authors.forename: V P
authors.initials: VP
authors.lastname: Rane
authors.forename: M L
authors.initials: ML
authors.lastname: Kubal
authors.forename: S
authors.initials: S
authors.lastname: Singh
authors.forename: M V
authors.initials: MV
authors.lastname: Patel
authors.forename: H F
authors.initials: HF
authors.lastname: Khorakiwala
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000191:economics / Q000379:methods*; D024841:Fluoroquinolones / Q000032:analysis*; D016106:Methicillin Resistance; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet; D013211:Staphylococcus aureus / Q000187:drug effects; D013237:Stereoisomerism; D047392:beta-Cyclodextrins / Q000737:chemistry
chemical_list: D024841:Fluoroquinolones; C490933:WCK 771; D047392:beta-Cyclodextrins