Abstract: OBJECTIVE To separate and identify the chemical constituents in Macrothele yani and determine the content.METHODS The volatile oils were extracted by steam distillation method and Soxhlet extraction, and the volatile oils were identified by GC-MS. Total amino acids content was determined by amino acid autoanalyzer and trace elements content was determined by ICP-MS. RESULTS A total of 22 compounds were identified from the volatile oil extracted by steam distillation, while there were 18 kinds of volatile oils extracted by Soxhlet extraction. There were 7 volatile oils in common. Among them, ethyl oleate, ethyl linoleate and ethyl octadecanoate were the most abundant. Among those amino acids, Macrothele yani had 16 kinds of amino acids, including 7 kinds of essential amino acids. The total amount of amino acids was 10.53%. The contents of glutamic acid and arginine were relatively high. It was rich in 11 kinds of essential trace elements for human body, among which the content of Zn, Fe and Mn was higher, the content of heavy metal elements As and Pb met the limit requirements, and the contents of Cd and Cu exceeded the limit. CONCLUSION The volatile oil extracted from Macrothele yani by the two methods has great differences in type and content, some of them have certain medicinal effects. In addition, there are many kinds of necessary amino acids and trace elements for human in Macrothele yani.