Abstract: OBJECTIVE To establish GC-MS fingerprint of the volatile oil in the classical famous prescriptions Wuyao decoction substance to analyze the chemical composition of the volatile oil in the substance benchmark, the multi-index content determination method of Wuyao decoction was established to construct its quality control system. METHODS According to the ancient books and combining with the previously inspected process, different batches of decoction pieces were fed in mixed batches to prepare the material standard of Wuyao decoction, and GC-MS was used to establish the material standard fingerprint. Similarity was calculated by using "TCM Chromatographic Fingerprint Similarity Evaluation Software(2012 edition)". Combining with orthogonal partial least squares discriminant analysis, the main components that affected the quality of Wuyao decoction substance benchmarks from different batches and origins was excavated. Use 4 of these landmark compounds (cyperotundone, α-cyperone, dehydrocostus lactone, ligustilide) as indicators and used as indicators for content determination of Wuyao decoction substance benchmark. Content determination methods were established and the content of 10 batches of samples was determined respectively. RESULTS The similarity between the fingerprints of the 10 batches of samples and the control chromatogram was 0.954 to 0.990. Identified 27 common peaks, identified 26 chemical components and find 8 indexes which influence the stability of batch between components (peak 6, 21, 22, 9, 23, 18, 27, 20). The contents (cyperotundone, α-cyperone, dehydrocostus lactone, ligustilide) in the substance benchmark of Wuyao decoction were 5.54%, 2.93%, 1.52%, 8.89%. CONCLUSION GC-MS fingerprint and major component content determination of preliminary established the Wuyao decoction substance benchmark quality control method of the volatile oil.