Estimation of Related Substances in Tigecycline by rp-hplc Method

Estimation of related substancesby usinghigh-performance liquid chromatographic method was developed and validated for the determination of Tigecycline in the present work. Reversedphase chromatography was performed on Waters 2489 UV 2695 pump, Waters 2998 PDA 2695 pump Software Empower2 photodiode array detector using Zorbax Eclipse plus C18 (100 mm × 4.6 mm, 1.8 μm particle size) column with eluent-A: pH 6.50 buffer: acetonitrile: DMSO (90:5:5 %v/ v/v) and eluent-B: pH 6.50 buffer: acetonitrile: DMSO (71:24:5 %v/v/v) as mobile phase at a flow rate of 1.0 mL/min. with UV detection at 270 nm.Linearity was observed in the concentration range of Tigecycline LOQ–1.13% (R2 = 1.000), the concentration range of di-MA-TIG impurity 0.04– 0.23% (R2 = 1.000), the concentration range of CMI 0.05-0.23% (R2 = 0.999). The limit of quantitation (LOQ) and limit of detection (LOD) were found to be di-MA-TIG impurity 0.0001 and 0.0004mg/ mL, CMI impurity 0.0001and 0.0004μg/mL, Tigecycline 0.0001and 0.0005mg/mL respectively. The method was validated as per ICH guidelines. The %RSD precision was found to be less than 1.0 %. The percentage recovery was in good agreement with the labeled amount in the pharmaceutical formulations and the method is simple, specific, precise and accurate for the determination of Tigecycline in pharmaceutical formulations.