ABSTRACT
OBJECTIVE:To establish a method for the determination of related substances in regorafenib. METHODS:Gradi-ent elution HPLC was performed on the column of Waters Atlantis T3 C18 with mobile phase A of 0.1%Trifluoroacetic acid solution and B of acetonitrile(gradient elution)at a flow rate of 1.0 ml/min,the detection wavelength was 232 nm and column temperature was 35 ℃,injection volume was 10 μl. RESULTS:AFP-PMA urease(impurity A),FP- dimethyl pyridine carboxamide(impurity B),3,4-bis-CTF-aminoformamido-PMA(impurity C),regorafenib and other impurities were well separated;the linear range was 0.511-5.108 μg/ml for impurity A(r=0.999 9),0.287-2.869 μg/ml for impurity B(r=0.999 5),0.360-3.604 μg/ml for impurity C (r=0.999 9)and 1.444-14.442 μg/ml for regorafenib(r=0.999 8);the detection limit were 0.052,0.022,0.084 and 0.071 μg/ml, respectively;RSDs of precision,stability and reproducibility tests were lower than 3%;recoveries of impurity A,B and C were 102.7%-106.3%(RSD=1.09%,n=9),98.2%-102.9%(RSD=1.83%,n=9)and 98.6%-104.3%(RSD=1.57%,n=9). CON-CLUSIONS:The method is sensitive,rapid,accurate and reliable,and can be used for the determination of related substances in regorafenib.