Pre-Column derivatization Chiral HPLC Method for the separation and quantification of (R,R)-2,8-diazobicyclo [4.3.0]nonane content in (S,S)-2,8-diazobicyclo[4.3.0]nonane, A Key Intermediate of Moxifloxacin Hydrochloride

A simple and robust analytical method was developed for the separation and quantification of (R,R)-isomer in (S,S)-2,8-diazobicyclo[4.3.0]nonane, a non chromophoric moiety and key intermediate in the synthesis of Moxifloxacin hydrochloride by using pre-column derivatization reagent 4-Chloro-7-nitrobenzofurazan (NBD Chloride). The derivatization was brought to be optimized at room temperature. The separation was achieved on Chiralpak IC 250 mm × 4.6 mm, 5 µm column with isocratic elution mode with a flow rate of 1.0 mL min-1. The mobile phase was a mixture of methanol, ethanol and diethyl amine in the ratio of 1:1:0.1 (v/v/v). The eluents were monitored at 340 nm using a UV detector.