Farfarae Flos (FF) was obtained from a local drug store and authenticated as the buds of Tussilago farfara L. by Prof. Zhenyu Li. Voucher specimens (No. KD-34) were deposited in the herbarium of the Modern Research Center for Traditional Chinese Medicine of Shanxi University. The preparation and chemical characterization of PEFF have previously been reported by us 20. In brief, the dried FF flower buds (3 kg) were ground to a powder and extracted three times for 60 min each with 4 L of petroleum ether (60-90℃) with ultrasonication. The combined extract was then concentrated under reduced pressure (yield 6.5 g of PEFF) and used for experiments.

In previous studies 20, the typical 1H NMR spectrum of PEFF was obtained. In the region of δ 4.5-6.5 and δ 0.8-1.2, the signals corresponding to the doubles bonds and methyl groups of the sesquiterpenoids were obviously detected. In addition, seven compounds were identified in PEFF by LC-MS analysis. Finally, we proved that the sesquiterpenoids were present as the major components in the PEFF.

The sesquiterpenoids are soluble in lipophilic organic solvents. Considering the low toxicity and excellent permeability of the organic solvent dimethylsulfoxide (DMSO), the lyophilized PEFF was dissolved using 20% DMSO/saline. Furthermore, based on the PEFF treatment concentration in OVA-induced rat asthma models 20, low dose (LD, 0.01 g/kg), medium dose (MD, 0.02 g/kg), and high dose (HD, 0.04 g/kg) of PEFF were used.

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