Tylosin tartrate was used as a target drug in this study to investigate the influence of petroleum ether extract from Platycodon grandiflorum roots on the pharmacokinetics and its distribution character in tissue of mice by oral administration. The concentrations of tylosin tartrate in mice plasma and tissue at different time were determined using high performance liquid chromatography. The pharmacokinetic data was calculated by Pkslover program and analyzed by SPSS19.0 software and non compartment model. The result showed that the curve of time-concentration from tylosin tartrate in blood exhibited multiple peaks, compared with control group, half-life time(t1/2)became shorter, peak concentration and peak time decreased, area under the curve(AUC)and mean residence time(TMR)decreased as well, however, numerical value of apparent volume of distribution (Vz) and clearance (Cl) increased; It was showed that the concentration of the drug in heart was in the highest among tissues. In general, petroleum ether extract from Platycodon grandiflorum roots could significantly influence on the pharmacokinetic parameters of tylosin tartrate in healthy mice, including on speeding up drug absorption, relieving the degree of absorption, diffusing drug in bodies, and accelerating its rate of clearance.