Study On The Anti Allergic Effect And Preparation Process Of Tussilagone

Objective① Extraction,separation and purification of Tussilagone from Tussilago.②Confirm the anti allergic pharmacological effects of tussilagone by choosing different mouse anti allergy models,and explore its mechanism.③ Prepare the Tussilagone nasal drops,and to establish the quality standards of the nasal drops(Draft).Methods① Isolation,purification and analysis of Tussilagone:extract,separate and purificate of Tussilago from tussilago farfara through the ultra supercritical CO2 extraction,silica gel column chromatography and HPLC preparation,and identifie it’s chemical structures.② Anti-allergic effects research:study on the anti-allergic effects of Tussilagone by delayed hypersensitivity reaction in mice and allergic rhinitis model of allergic mouse(Intranasal administration,oral administration).③ Tussilagone nose drops agent preparation:the prescription of nasal drops was determined by the single factor investigation of the types and dosage of emulsifier,thickener,pH regulator and preservative.Prepare three batches of samples of Tussilagone nasal drops according to the prescribed preparation.④ Tussilagone nasal drops’ quality standards:According to the 2010 edition of Chinese Pharmacopoeia,observe the nasal drops’ appearance,identificate Tussilagone through TLC,analysis Tussilagone’s content through HPLC and determine the pH value.⑤The preliminary stability test:Investigated the the effects of temperature,humidity,light and other external factors on the characteristics,identification,content determination,pH value of nasal drops,and compared the results with the zero month.Results① Active fraction Kdj4 was separated from Tussilago farfara L.through supercritical extraction,which accounts for 1.39%.Tussilagone was isolated from Kdj4 by silica gel column chromatography and HPLC preparation which purity is 98.14%.The total yield was 0.04%.②High,medium and low dose group of tussilagone all had a certain inhibitory effect on ear edema in mice which was caused by delayed type hypersensitivity.The inhibition rate is 51.78%、48.59%and 21.87%respectively,present a dose-response relationship.The high dose which decreased mouse ear thickness and swelling significantly inhibition effect was similar with positive drug prednisone.③High and medium dose group of tussilagone can significantly inhibited allergic rhinitis caused by OVA through gastric perfusion,reducing sneezing、scratching the nose,runny nose etc.The serum test results showed that high and medium dose group of tussilagone can decrease histamine and NO content.④High and medium dose group of tussilagone can significantly inhibited allergic rhinitis caused by OVA by intranasal.The inhibitory effect of high dose group was similar to loratadine while the medium dose group had obvious inhibitory effect on behavior and release of NO and histamine in serum of allergic rhinitis mice.⑤Preparation of prescription of tussilagone nasal drops:Tussilagone 460mg;Tween-804mL;some plant oils;CMC-Na 2.5g;ethylparaben 0.1g,adjust pH value to 6.0,produced to 100mL,made of 10 tubes.⑥ Tussilagone nasal drops’ quality standard:This product was milky white homogeneous liquid preparation.per tube contained Tussilagone 41.1mg;pH 6.0 or so..⑦ The Tussilagone nasal drops showed good stability from either the appearance or the intrinsic quality.Conclusion① The extraction and preparation of Tussilagone technology was simple,reasonable and stable.② The main type of Tussilagone anti-allergic was type I mouse allergic rhinitis and type Ⅳdelayed hypersensitivity reaction.③ Tussilagone’s anti allergic effects mainly related to the inhibition of NO and histamine mediators.④ The manufacture method of Tussilagone nasal drops was process reasonable,quality controllable and stable.