A simple and rapid method was developed for the determination of disophenol in bulk drug and injection via high performance liquid chromatography（HPLC） with the internal standard method.The sample was prepared with methanol to the test concentration,and filtrated with microfiltration membrane.Separation of analytes was achieved under isocratic elution on a C18 column.The mobile phase consisted of a mixture of 0.02 mol/L potassium dihydrogen phosphate（pH 2.6） and methanol（30∶70）,and was delivered at a flow rate of 1 mL/min.The UV detection was set at 235 nm,and the column temperature was maintained at 30 ℃.The internal standard was nitroclofene and the injection volume was 20 μL.Under the optimum experimental conditions,the proposed HPLC-internal standard method provided a good linearity in the range of 20-120 mg/L for disophenol with the correlation coefficients（r） of 0.999 6.The average recoveries of disophenol at three spiked concentration levels of 64,80 and 96 mg/L were in the range of 90.4%-98.2%,with relative standard deviations（RSDs） of 0.18%-0.72%.The limits of quantitation（LOQs,S/N=10） and limits of detection（LODs,S/N=3） were 1.5 mg/L and 0.15 mg/L,respectively.With good precision,stability and repeatability,the established method was simple,accurate,rapid and reliable for the determination of disophenol in bulk drug and injection.