| Objective: The Adiantum capillus-veneris L. as the research object, establishing a qualitative and quantitative method for the total flavones from Adiantum capillus-veneris L.(TFAV) and optimize its extraction and purification technology; Studing the acute toxicity, anti-inflammatory, antitussive, expectorant and antiasthma effects of TFAV, aimed at providing scientific basis for its clinical application. Methods: 1.color reaction and thin layer chromatography(TLC) methods were used to identify the TFAV, and studied the methodology. 2. The quantitative analysis method of TFAV was established with the method of Ultravoilet Spectrophotometry(UV) with rutin as standard sample and NaNO2-AI(NO3)3-NaOH as color-developing reagent; Comparing four extraction methods, based on the result, the method of ethanol extraction using single factor experiment and orthogonal test was used in the extraction of TFAV with the extraction rate of flavonid as index. Followed the content of TFAV were determined. 3. With the content of TFAV as index, using the combining techniques of macroporous and polyamide resin, adsorption and elution behavior of polyamide resin for TFAV were studied and selected optimization technique parameters for purification technology of TFAV. 4. Acute toxity effects was determined by maximum dosage method.5. Using methods such as ear edema induced by xylene, cough induced by ammonia, tracheal phenol red extraction and asthma induced by spaying the whole experiment animals observe the anti-inflammatory, antitussive,expectorant and antiasthma effects of TFAV. Results: When developing agent was ethylacetate-formic acid water(10:1.5:1.5), rutin was determined and the spots was clear with good separation. 2.The UV method had a good linear relationship in the range of A=11.411C+0.0015, average recovery was 98.77%. The optimum extraction conditionsof TFAV were as follows: ethanol concentration 50%, extraction temperature 80℃, the ratio of solid to liquid 1:25, extraction time 2.0h. The residues were extracted in duplicate2 times. The results of variance analysis showed that the major factor affecting the extraction of the TFAV is the concentration of extraction time, followed extraction temperature, then the ratio of solid to liquid, finally ethanol concentration. Under the optimal condition, the extraction rate of the TFAV could reach 3.14%. 3. Optimum purification technology parameters were as follows: using 60-100 mech polymide, sample volume of 24.8331mg·g-1, concentration of sample solution 5.4126 mg·g-1, the adsorption and elution velocity of 1.0 mL·min-1, eluted impurities with 3BV of water and 3BV of10% ethanol, then eluted with 3BV of 70% ethanol, collected eluent of the 2-6BV, purity of total flavones were up to 51.01% from 3.14%. 4. No acute toxicity was observed in the control group. The maximum dosage of TFAV by ig in mice was 40 g·kg-1, which was about 160 times of adult chemical daily dose(15g). 5. Compared with the control group,TFAV could decrease ear swelling and the frequency of cough(P<0.05), prolong the latent period(P<0.05), promote tracheal phenol red extraction(P<0.05) and make the asthma latency prolonged(P<0.05). Conclusion: The qualitative and quantitative methods were easy operated and reproductive. The results initially show that the crude extracts of Adiantum capillus-veneris L. contain flavones. Extraction and purification effects of TFAV were feasible and with low cost. It could enrich the total Flavones effectively. TFAV has the significant anti-inflammatory, antitussive, expectorant and antiasthma effects and safe to be a certain extent. |
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