Analysis Of Main Effective Components In Tricyrtis Maculata Machride

This study was based on Tricyrtis maculate Machride belonging to Tricyrtis Wall of Liliaceae,which is one of Chinese traditional Chinese used frequently in Shaan Xi province.This paper mainly analyzes and studies the active ingredients of Tricyrtis maculate Machride.The main research contents are:?1?The content and accumulation of total flavonoids in T.maculate Machride;?2?Optimization of the extraction technology of total flavonoids in T.maculate Machride by response surface method;?3?Simultaneous determination of rutin,kaempferol 3-rutinoside and kaempferol in T.maculate Machride by HPLC;?4?Antioxidant activities in vitro of different solvent extracts from T.maculate Machride;?5?Bacteriostatic activities of different solvent extracts from T.maculate Machride.The main research results are as follows:1.Studies on the analysis of content and accumulation of total flavonoids in T.maculata MachrideThe ultrasonic assisted extraction method was established and the content and accumulation of total flavonoid in different growth stages of different parts were determined by ultraviolet spectrophotometry with NaNO2-Al?NO3?3-NaOH color-developing system.The results showed that rutin has good linearity in the range of 3.1776?g/mL-79.44?g/mL,and the average recovery is 100.66%in RSD of 2.17%?n=6?.The total flavonoid content in different growth stages of different parts of T.maculate Machride has significant difference and the total flavonoid content reach the highest point in September with the value of 27.147mg/g.So that,the best harvest time for T.maculate Machride.was in September Those results provided a theoretical reference and experiment basis for the medicinal parts chosen and optimum harvest phase of T.maculate Machride.2.Optimization of extraction technology of total flavonoids from T.maculata Machride by response surface methodAn ultrasonic procedure for the extraction of total flavonoids from T.maculate Machride was optimized by using Box-Benhnken center-united experiment design and response surface analysis method.The main ultrasonic extraction factors including extraction ultrasonic power,extraction time,extraction temperature,solid-liquid ratio which effect the extraction yield of total flavonoids from T.maculata Machride were analyzed.The results showed that factors affecting extraction yield of flavones from T.maculate Machride in sequence were extraction ultrasonic power,solid-liquid ratio,extraction temperature,extraction time and the optimum extraction conditions of total flavonoids were as follows:Extraction temperature of 45?,material liquid ratio of 1:40?g:mL?,extraction time of 60 min,extraction ultrasonic power of210 W.The extraction yield of total flavonoids was 12.966 mg/g under the optimal extraction conditions,which was nearly same to the theoretical highest yield of 13.035 mg/g predicted by the response surface analysis method.3.Simultaneous determination of rutin,kaempferol 3-rutinoside and kaempferol in T.maculata Machride by HPLCThe method of simultaneous determination of rutin,kaempferol 3-rutinoside,kaempferol in T.maculata Machride by HPLC was established and the content and accumulation of flavonoids in different growth stages from T.maculate Machride was determined.The chromatographic separation was caried out by using Inertsil ODS-3C₁₈ column?4.6mm×150mm,5?m?,the mobile phase was methanol:0.3%phosphoric acid,the flow rate was 0.8mL/min,the column temperature was 30?,and the detection wavelength was set at 360 nm.The results showed that the biggest yield of rutin in T.maculata Machride is 916.692?g/g,which of Kaempferol 3-rutinoside was 645.586?g/g and that of kaempferol was 14.924?g/g.The content of flavonoids in different growth stages had significant difference and the yield of rutin was the greatest,which was followed by that of kaempferol and the kaempferol 3-rutinoside was the least all the time.the yield curves of Rutin and kaempferol 3-rutinoside of flavone glycosides presented M-type form,and that of kaempferol of flavonoid aglycone presented the V-type form.4.Antioxidant activities in vitro of different solvent extracts from T.maculate MachrideActive materials of T.maculate Machride was extracted by water,methanol,50%ethanol and antioxidant activities of different solvent extracts was determined respectively by phenanthroline,DPPH and Fe³⁺reducing power.The results showed that antioxidant activities of different solvent extracts from T.maculate Machride were difference,and a positive correlation between concentration and antioxidant activity was found just as the higher the concentration was,the stronger the antioxidant activity would be.Antioxidant abilities of T.maculate Machride in sequence were 50%ethanol,methanol,water,which,however,were all inferior to Vc's antioxidant capacity.5.Bacteriostatic activity of different solvent extracts from T.maculate MachrideThe inhibition activities of different solvent extracts from T.maculate Machride on Staphylococcus aureus,Escherichia coli,Pseudomonas aeruginosa and Monilia albican was determined by filtering paper method.The results showed that the 50%ethanol extract had bacteriostatic activity on Escherichia coli and not to other target bacteria,while the water extract and methanol extract didn't.The results of this experiment fill the gaps in research field about bacteriostatic activity of T.maculate Machride and provideed scientific reference for the biological activity of T.maculate Machride.