Objective To develop a method for the determination of ganciclover in human plasma by RPHPLC.Methods Plasma containing ganciclover was extracted with methanol and methylene chloride,qualitative and quantitative analysis was carried out directly.Working curve,linear range,recovery,precision and so on was obtained according to the sample pre-processing method and analysis state.The HPLC method has been taken to investigate the plasma concentration of ganciclover for 12 volunteers.Results The relationship of the peak area of ganciclover concentration in plasma linear within the range of 0.05 μg/mL～1.60 μg/mL(r=0.9999).The lowest detection limit was 0.01 μg/mL(S/N≥13).The intra and inter-day RSD were less than 5.1%respectively.The recovery is about 90.0%～95.4%.Conclusion The established method in the article was shown to be sensitive,accurate and simple for the determination of ganciclover level.It is suitable for clinical detection of ganciclover and forensic medicine and toxicology analysis.