Abstract: Matrix solid-phase dispersion-gas chromatography combined with mass spectrometry(MSPD-GC/MS) analysis method for essential oil was established based on Hainan agarwood induced by biotechnological method. The MSPD conditions of extracting agarwood essential oil established by single factor test were magnesium silicate as the adsorbent, adsorbent/sample mass ratio 4:1, isopropanol as the dispersion solvent, dispersion solvent/sample ratio 4:1(mL: g) and 6 mL n-hexane/ethyl acetate 9:1(volume ratio) as the elution solvent. Under these conditions, the extraction yield of MSPD was (9.19±0.15)%, which was higher than ultrasonic assisted ether extraction (6.65±0.22)%. The chemical constituents of essential oil were analysis by GC/MS, and compared with that extracted by ultrasonic assisted ether extraction. The compositions of essential oil extracted by MSPD were similar to ultrasonic assisted ether extraction, the main constituents were 2.13% aromatic, 25.67% sesquiterpenoids, 26.85% chromones and 7.50% aliphatic compounds. The number of sesquiterpenoids in the essential oil extracted by MSPD was more than that of ultrasonic assisted ether extraction, such as γ-selinene, β-santalol, valerena-4, 7(11)-diene, α-guaiene, shyobunol and 1, 9-aristo-ladiene. Thus, the MSPD-GC/MS analysis method could truly reflect the chemical composition of agarwood essential oil, which could be used as a rapid identification method for the quality evaluation of agarwood.