HPLC法测定瑞戈非尼原料药中的有关物质

Determination of Related Substances in Regorafenib by HPLC

目的：建立测定瑞戈非尼原料药中有关物质的方法。方法：采用高效液相色谱法。色谱柱为WatersAtlantisT3C18，流动相为0.1％三氟乙酸-乙腈（梯度洗脱），流速为1.0ml/min，检测波长为232nm，柱温为35℃，进样量为10μl。结果：AFP-PMA脲（杂质A）、FP-二甲基吡啶甲酰胺（杂质B）、3，4-双-CTF-氨基甲酰胺基-PMA（杂质C）与主成分瑞戈非尼及其他未知杂质均能达到很好的分离；杂质A、B、C和瑞戈非尼检测质量浓度线性范围分别为0.511～5.108μg/ml（r＝0.9999）、0.287～2.869μg/ml（r＝0.9995）、0.360～3.604μg/ml（r＝0.9999）和1.444～14.442μg/ml（r＝0.9998）；检测限分别为0.052、0.022、0.084和0.071μg/ml；精密度、稳定性、重复性试验的RSD＜3％；杂质A、B、C加样回收率分别为102.7％～106.3％、98.2％～102.9％、98.6％～104.3％，RSD分别为1.09％、1.83％、1.57％（n＝9）。结论：该方法灵敏、快速、准确、可靠，可用于瑞戈非尼原料药中有关物质的测定。

OBJECTIVE:To establish a method for the determination of related substances in regorafenib. METHODS:Gradientelution HPLC was performed on the column of Waters Atlantis T3 C18 with mobile phase A of 0.1% Trifluoroacetic acid solutionand B of acetonitrile(gradient elution)at a flow rate of 1.0 ml/min,the detection wavelength was 232 nm and column temperaturewas 35 ℃,injection volume was 10 μl. RESULTS:AFP-PMA urease(impurity A),FP- dimethyl pyridine carboxamide(impurityB),3,4-bis-CTF-aminoformamido-PMA(impurity C),regorafenib and other impurities were well separated;the linear range was0.511-5.108 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 were102.7%-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). CONCLUSIONS:The method is sensitive,rapid,accurate and reliable,and can be used for the determination of related substances inregorafenib.