OBJECTIVE： To establish a method for the determination of related substances in Cabazitaxel injection. METHODS： HPLC method was used. The determination was performed on Agilent Eclipse XDB-C18 column with mobile phase consisted of water-acetonitrile-ethanol （gradient elution） at the flow rate of 1.0 mL/min. The column temperature was 35 ℃， and the detection wavelength was set at 230 nm. The sample size was 20 μL. Established method was used to determine related substances in 3 batches of Cabazitaxel injection. RESULTS： The linear relationship of cabazitaxel were 0.039-11.60 μg/mL （r＝0.999 8， n＝7）. The detection limit was 2×10－4 μg， and quantitation limit was 8×10－4 μg. RSD of precision and reproducibility tests were all lower than 10.0％ （n＝6）. The amount of single impurity in 3 batches of samples ranged 0.07％-0.08％， and total amount of impurities were 0.26％-0.29％. CONCLUSIONS： Established method is simple， accurate and reliable， can be used for the determination of related substances in Cabazitaxel injection.