Development and Validation of HPLC Method for determination of Decitabine impurity profile in Decitabine for Injection 50mg/vial

Decitabine is an anti-cancer chemotherapy drug. This article describes method development and method validation of related substances of Decitabine (α- decitabine, α-isomer, methyl 4-chlorobenzoate, β-isomer, O-Acetyl and unknown impurities) in decitabine drug substance and finished dosage forms by using a high Performance liquid chromatography. In this high performance liquid chromatography, the resolution was achieved on Inertsil ODS 3V, 250×4.6 mm, 5μm column with a gradient elution at a flow rate of 1.2 mL/min using a mobile phase A as Ammonium acetate buffer solution and mobile phase B as water: acetonitrile (10:90% v/v) at wavelength 254 nm by an UV detector. The method was validated in the concentration range of 0.7 ppm to 7.1 ppm, 1.2 ppm to 11.0 ppm, 1.2 ppm to 10.9 ppm, 1.2 ppm to 11.0 ppm, 1.2 ppm to 10.9 ppm and 0.8 ppm to 7.7 ppm for related substances of α- decitabine, α-isomer, methyl 4-chlorobenzoate, β-isomer, O-Acetyl and Decitabine respectively. The obtained recovery was in between 90.0 % to 110.0 % and the % RSD was not more than 10.0. The HPLC method is suitable, specific, linear, accurate, precise, stability indicating and robust which may be useful for the routine and stability analysis for determination of related substances in Decitabine drug substance and finished dosages.