Objective:To develop an LC-MS/MS method for determination of sofalcone in urine samples,and to investigate its urinary excretion characteristics in healthy Chinese volunteers. Methods:The urine samples were treated with methanol and separated on a Poroshell 120 SB-C18 column,with a mobile phase of methanol and 2 mmol/L Ammonium acetate(85∶15,v/v). The rate was 0.20 mL/min and the temperature was 30℃. Electrospray ionization source was applied and operated in the negative ion mode using SRM. Two healthy Chinese volunteers were given single oral dose of sofalcone capsule(200 mg) for urinary excretion and characteristics. Results:The lower limits of quantitation for sofalcone were 1 ng/mL,and the linear range was 1~500 ng/mL. The data of intra and inter-run precision were all less than 6%. The matrix effects were between 93.1% and 114.8%. The extraction recovery was 84.7%~93.6%. The accumulated excretion rates of sofalcone in urine were lower than 0.01%. Conclusion:The developed method can be applied to evaluate the urinary excretion of the sofalcone with a very short running time.