A method using HPLC is established to determine orientin in tissue of rabbits and to study the characteristic of the orientin when injected into veins. After orientin is injected into veins, tissue samples are collected at time sharing. The rabbit is put to death by bloodletting, then the heart, liver, brain, kidney, lung and spleen are taken out to prepare homogenate tissue. HPLC is used for determination of orientin in tissue of rabbits. Theory of statistical moment is used to account AUC of each tissue sample. Endogenous material in tissue does not interfere in determination of orientin. In amplitude of 0.0500～50.0mg/L, mass concentration of orientin and peak area have a good linear relation, and the quantitation of lower limit is 0.0500mg/L. The RSD of precision,accuracy and plus sample recovery are all less than 15%. The AUC order of each tissue is kidney＞liver＞lung＞spleen＞heart＞brian.After log transition of AUC data, normality test, homogeneity test for variance and analysis of variance are conducted.The result shows AUC of each tissue have significant difference(p＜0.05).Orientin distribute in dominating tissue of rabbits is obviously different.