Simultaneous determination of related substances in entecavir film-coated tablet by high performance liquid chromatography.

: To establish a high performance liquid chromatography method to simultaneously quantify eight related substances in entecavir film-coated tablets.According to USP40 and YBH33292005 standards, a high performance liquid chromatography (HPLC) method for simultaneous determination of 8 related substance in entecavir film-coated tablets was established and validated. The column was WATERS C18 (250 mm the mobile phase A was water-acetonitrile-trifluoroacetic acid (990∶10∶1), the mobile phase B was water-acetonitrile-trifluoroacetic acid (700∶300∶1) with gradient elution and ultraviolet-visible light detector, detection wavelength at 254 nm and column temperature of The resolution between entecavir and each impurity peak was more than 1.5, and each impurity had a good linear relationship with the peak area in the linear range. The limits of detection and quantification, precision, stability, durability, specificity, met the verification requirements. The HPLC method established in this study can be used for simultaneous determination of 8 related substance in entecavir film-coated tablets.