OBJECTIVE: To conduct component analysis for the 40% ethanol eluate of Hyssopus officinalis, and investigate its improvement effect on inflammation in asthmatic mice. METHODS: The 40% ethanol eluate of polyamide resin column was collected, and HPLC-high resolution mass spectrometry was used for the component analysis of 40% ethanol eluate of H. officinalis. Totally 72 mice were randomly divided into blank group (normal saline), model group (normal saline), dexamethasone group (positive control, 1.6 mg/kg) and 40% ethanol eluate of H. officinalis high-dose, medium-dose, low-dose groups (200, 100, 50 mg/kg), 12 in each group. Except for normal group, mice in other groups were intraperitoneally injected 0.2 mL ovalbumin (OVA) for sensitization in 0, 14 d and intragastrically administrated in 25-31 d, once a day. After administration, 2 mg/mL OVA was dropped in nose for 7 d. After 24 h of last dropping in nose, tumor necrosis factor α (TNF-α), interleukin-4 (IL-4), interferon-γ (IFN-γ) levels in bronchoalveolar lavage fluid (BALF) were detected; pathological changes in lung tissue were observed. RESULTS: Totally 11 compounds were identified, the relative percentage content of 40.89%. The main components were rosmarinic acid, luteolin 7-O-β-D-rhamnoserhamnose (1→6)-α-D-pyran glucoside, hyperoside, etc. Compared with blank group, TNF-α, IL-4 levels in BALF in model group were increased, IFN-γ level was declined, and IL-4/IFN-γ ratio was enlarged (P<0.01); lung tissue was seriously damaged, there was infiltration of inflammatory cells around the blood vessels. Compared with model group, TNF-α, IL-4 levels in BALF in dexamethasone group, 40% ethanol eluate of H. officinalis high-dose, medium-dose groups were declined, IFN-γ level was increased, and IL-4/IFN-γ ratio was reduced (P<0.01); pathological changes in lung tissue were improved. CONCLUSIONS: The established analysis method can effectively analysis the chemical components of 40% ethanol eluate of H. officinalis, which has certain regulatory effect on releasing inflammatory factors and reducing inflammatory lesions in lung tissue of mice with bronchial asthma.