| The traditional Chinese medicine rhizoma sparganii belongs to the sparganiaceae,which is dry tuber of the inner Sparganium stoloniferum Buch.-Ham.The medicinal material tastes bitter and the medical property is mild.It is mainly attributed to the two meridians as liver and spleen,which has the function of promoting blood circulation and relieving swelling and pain.In the medical field,this medicine is used for the treatments such as gynecological dysmenorrheal,pain of hearth attack and distending pain of stomach.The main components of rhizoma sparganii include flavonoid,phenylpropanoids and volatile oils.Among them,the chemical component of flavonoids is considered as one of the main medicinal ingredients of rhizoma sparganii,which has prominent effect in aspects of antithrombus,anti-platelet aggregation and anti-tumor,and plays an important role in the efficacy of promoting flow of qi and blood circulation.Based on the previous research,this paper investigates the extraction process and purification process of the total flavonoids of rhizoma sparganii,determines the preparation process of the total flavonoids of rhizoma sparganii,and preliminarily estimates the structure of the compounds contained in it by liquid chromatography mass spectrometry technology.In addition,this paper systematically studies the chemical components of 70% ethanol extract of rhizoma sparganii,so as to provide material basis for the further study on the drug efficiency.The experiment is divided into four parts,which are described as follows:In the First part: the rutin is used as a reference substance to draw the standard curve for the measurement of the total flavonoids of rhizoma sparganii.Its precision,stability,repeatability and recovery rate are all good.The extraction rate of total flavonoids from the rhizoma sparganii is used as an indicator to investigate the single factors of ethanol concentration,extraction time,power-baking particle size of the medicine rhizoma sparganii,and feed liquid.Then,through the orthogonal design experiment,the final extraction process of the total flavonoids of rhizoma sparganii is confirmed as: use 40 orders of coarse powder of medicinal material;70% ethanol dosage is 8 times the amount of medicinal materials;extract for 2 times with 1.5h each time.According to the verification experiment,it can be known that the process has good stability and reproducibility.In the Second part: the macroporous resin is used to purify the total flavonoids of rhizoma sparganii.The yield of the total flavonoids of rhizoma sparganii after upper prop is regarded as the indicator.The most suitable model for the purification of the total flavonoids of rhizoma sparganii is selected from 7 different models of macroporous resins;then the diameter height ratio,the loading concentration,the loading p H value,the loading flow rate,the effluent volume,the effluent concentration and the effluent flow rate are separately investigated by single factor.Finally,according to the central combination experiment design principle,the designexpert software is used to select the three factors as loading p H value,effluent concentration and effluent flow rate which have the greatest impact on indicator for the response surface test.Then the optimal solution for the macroporous resin purification process of the total flavonoids of rhizoma sparganii is: 2*50cm glass column.The wet packing method is used for the macroporous resin HP20 with the column diameter height ratio of 1:7.The 50 m L0.4g/m L of rhizoma sparganii extract with p H = 4.8 is loaded at a speed of 0.5BV/h.After complete adsorption,the distilled water is used for elution till the effluent is nearly colorless.Then the elution is carried out with 100 m L72 % ethanol at a speed of 2.0 BV/h.the purity of total flavonoid is increased from 4.25%(stock solution)to 40.83%(RSD=2.56%,n=3).The total flavonoid yield is 72.34%(RSD=1.77%,n=3).Compared with the 73.99% of predicted model value,the relative error is 2.13%.In the third part,To study the chemical constituents of Sparganium stoloniferum Buch.-Ham.The compounds were separated and purified by silica gel,Sephadex LH-20,MCI column,C8 column,ODS column chromatography and semi-preparative high performance liquid chromatography,and the 17 compounds were isolated and identified as 1,3-O-diferuloylglycerol(1)、1-O-feruloyl-3-O-P-coumaroylglycerol(2)、ferulicacid(3)、P-hydroxylcinnamic acid(4)、vanillic acid(5)、Phydroxybenz-aldehyde(6)、5-hydroxymethyl-2-furaldehyde(7)、β-sitosterol palmitate(8)、daucosterol palmitate(9)、β-sitosterol(10)、daucosterol(11)、formononetin(12)、rutin(13)、kaempferol(14)、succinic acid(15)、6,7,10-trihydroxy-8-octadecenoic acid(16)、2,3-dihydroxypropyl palmitate(17).In the fouth part,To study the chemical constituents of Sparganium stoloniferum Buch.–Ham by HPLC-MS,infer it have:Octan-3-ol、Ethyl pentanoate 、4-methoxy-3-methyl-6-[(1E)-1-methylprop-1-en-1-yl]-2H、3-(4-(tertButyl)phenyl)-2-methylpropanal、1,3-Dihydroxyacetone、Terephthalic acid、Oxacyclohexadecan-2-one、Succinic acid. |
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