Determination of related substances in apixaban tablets by HPLC-principal component self-control with correction factor
Objective To establish an HPLC-principal component self-control with correction factor method to simultaneous determine 12 related substances(impurities A～L)in apixaban tablets and their limit values.Methods The determination was performed on a Waters Xbridge C18 column(250 mm×4.6 mm,5 μm).The mobile phase consisted of 30 mmol·L-1 ammonium acetate buffer solution(pH 4.50)(phase A)and acetonitrile(phase B)with gradient elution at a flow rate of 1.0 mL·min-1.The column temperature maintained at 40 ℃,and the detection wavelength was set at 280 nm.The injection volume was 10 μL.The linear curve of apixaban and the 12 impurities were drawn and the correction factors of each impurity related to apixaban were calculated by slope.The genotoxicity of the impurities was predicted by Nexus 2.6.0 software.Results Apixaban and the 12 impurities were well separated.The linearity of apixaban and 12 impurities was good within the corresponding ranges(r＞0.999).The limit of quantification was 0.0396～0.0582 μg·mL-1.The average recoveries of low,medium and high concentrations(n=3)of impurities A～L were 101.77％～102.60％,101.82％～103.32％,100.10％～102.09％,99.55％～101.31％,99.91％～103.12％,100.36％～102.93％,100.86％～102.03％,101.25％～102.44％,103.13％～104.58％,100.33％～100.84％,100.17％～102.65％and 100.46％～103.03％,respectively.The impurity content measured by principal component self-control with correction factor method and the external standard method showed no significant difference(P＞0.05).The predicted genotoxicity of all the impurities by Nexus 2.6.0 software was negative(ICH M7 class 5).Conclusion This method is sensitive,specific and accurate,which can be used for the determination of related substances in apixaban tablets.
apixabanrelated substancehigh performance liquid chromatographyprincipal component self-control with correction factor
万方数据
维普
