Objective To explore a method of preparing the acellular tendons of human with chemical approaches.Methods From April 2011 to June 2012,several treatments were performed on human tendons using 1.00％ tri(n-butyl) Phosphate (TnBP) combinded with Triton X-100 at the concentration of 0,0.25％,0.50％,1.00％ respectively.Specimens were examined using histological observation,scanning electron microscopy,biomechanical testing and hydroxyproline quantitation.The results were then compared with fresh tendons of control group,so as to value the characteristics of decellularized human tendon scaffold.Results Acellular human tendons had glossy surface,intact aponeurotic membrane and satisfactory flexibility.A small number of disrupted cells remained in the 1.00％ TnBP + 0％ Triton X-100 treated tissue,while other three experimental groups successfully eliminate all cells.Intact and regular collagen architecture was retained in 1.00％ TnBP +0.25％ Triton X-100 treated tissue.1.00％ TnBP + 0.50％ Triton X-100 and 1.00％ TnBP + 1.00％ Triton X-100 treated tissue were nearly identical to 1.00％ TnBP +0.25％ Triton X-100 treated tissue,but the interval of collagen was slightly wider than the control group,the maximum load (385.22 ± 80.32N,398.22 ± 127.20N),ultimate tensile strength(46.69 ± 16.30Mpa,46.20 ±5.52Mpa) and hydroxyproline content(0.282663 ± 0.0110109 μg/mg,0.279355 ± 0.0102129 μg/mg) were statistically lower (P ＜ 0.05) compared with those of the control group,maximum load(533.28 ± 135.77N),ultimate tensile strength (65.56 ± 14.40Mpa) and hydroxyproline content (0.292882 ± 0.0100988 μg/mg) respectively.Conclusion The decellularization treatment with 1.00％ TnBP + 0.25％ Triton X-100 could be optimized for preparing acellular human tendons.