Determination of related substances in cangrelor by HPLC

WANG Qun; SUN Kai-li; SHI Bing-yu; LIU Jing-yan; CHEN Yu-jie

Abstract：Objective:To establish an HPLC method for determination of the related substances in cangrelor.Methods:Waters Symmetry C18 column (250 mm × 4.6 rm,5 μm) was chosen for the analysis;the mobile phase was 15 mmol· L-1 ammonium phosphate sodium perchlorate solution (adjusted to pH 7.0 with phosphoric acid)-acetonitrile with gradient elution at the flow rate of 1.0 mL· min-1,the detection wavelength was set at 242 nm,and the column temperature was 30 ℃.Results:The study included four main known impurities A,B,C and D in cangrelor,whose relative response factors were 0.590,0.510,2.27,0.660.According to this,an external stardard method using impurity reference substance was established to analyse the four impurities quantificationally,while an area normalization method was established to control the limit of unknown impurity.Chief peak and every impurity peak were separated well.Limit of Detection of A,B,C and D met the standard.Limit of Quantity of A,B,C and D were 1.00,0.048 0,0.025 0,0.075 0 μg·mL-1 and there was a good linearity separately over the ranges 1.00～8.50,0.0480～1.50,0.0300～1.50,0.0750～7.50μg·mL-1 The average recovery (n=9) of impurity A,B,C and D was included in 90％～ 110％,with RSD all less than 5.0％.Conclusion:After methodology validation,the method can be used not only for quantitative analysis of process and degradation impurities including A,B,C and D,but for limit control of unknown impurity in cangrelor as well.