Abstract: OBJECTIVE To identify the blood components of Modified Tubson-2, and to elucidate its mechanism of treatment of osteoporosis combine with network pharmacology. METHODS HPLC-Q-Exactive-MS/MS was used to identify the blood components of Modified Tubson-2; on this basis, the target prediction of blood components was conducted with SwissTarget Prediction and SuperPred database. At the same time, DisGeNET database was used to search for disease targets related to osteoporosis. Cytoscape software was employed to build a "blood component-target-disease" network model. Then String database platform was used to perform protein interaction network analysis. And David database was used for GO enrichment analysis and KEGG pathway analysis of core targets. Finally, the results of network pharmacology were validated by using Autoduck software. RESULTS Finally, 21 blood components were determined. Eleven core targets of Modified Tubson-2 anti-osteoporosis were obtained through network pharmacological analysis. According to the results of GO function annotation and KEGG pathway analysis of 11 core targets, Modified Tubson-2 treating osteoporosis mainly involved in the HIF-1, estrogen, MAPK, thyroxine, TNF, mTOR, PI3K/AKT signaling pathways. CONCLUSION This method preliminarily clarifies the blood components and potential mechanism of the Modified Tubson-2, and provides an important reference for further research on the pharmacological mechanisms of Modified Tubson-2.