Abstract: In order to clarify material basis of effective parts of Liangxue Tongyu prescription, blood-heat and blood-stasis rat model induced by dry yeast was established. The changes of rectal temperature, blood viscosity and plasma viscosity were used to evaluate the cooling-blood and activating-blood effects of Liangxue Tongyu prescription and its parts. Compared with the model group, the extract from Liangxue Tongyu prescription, its volatile oil and n-butanol part could significantly reduce rectal temperature (P < 0.01), and also reduce blood viscosity and plasma viscosity to various degrees (P < 0.01 or P < 0.05). So volatile oil and n-butanol part were primarily identified as effective parts of Liangxue Tongyu prescription. By using GC-MS with normalization method of area to analyze volatile oil of Liangxue Tongyu prescription, 70 compounds were identified, accounting for about 92.54%, mainly as β-asarone, paeonol, α-asarone and shyobunone. 42 compounds such as peony glycosides, tannins, and iridoid glycosides were identified by HPLC-MS techniques and standard comparison. The study determined the effective parts of Liangxue Tongyu prescription and clarified the chemical composition providing the foundation for further studies on material basis of Liangxue Tongyu prescription.