Study On The Quality Evaluation And Antioxidant Effect Of Gualou Seeds

Purpose:1.By observing the powder characteristics of in seeds of Fructus Trichosanthis,the stone cells in in seeds of Fructus Trichosanthis were selected as the characteristic cells,and the microscopic quantitative method of in seeds of Fructus Trichosanthis was established.The microscopic characteristic constants of in seeds of Fructus Trichosanthis were determined.2.By optimizing the extraction process of total triterpenoids from in seeds of Fructus Trichosanthis,the optimum extraction conditions ofin seeds of Fructus Trichosanthis were selected.On this basis,the contents of total triterpenoids in seeds of Fructus Trichosanthis from 10 different producing areas were determined,which provided a reference for more accurate determination of total triterpenoids in seeds of Fructus Trichosanthis.3.High performance liquid chromatography(HPLC)was used to establish the fingerprint of in seeds of Fructus Trichosanthis from 10 producing areas,and to determine the content of the specific components of in seeds of Fructus Trichosanthis,Pericarpium trichosanthis and Tricholoma,so as to establish the quality evaluation system of in seeds of Fructus Trichosanthis systematically in both qualitative and quantitative aspects,providing the basis for the quality control of in seeds of Fructus Trichosanthis.4.By using SPSS statistical software,the correlation between the content of total triterpenoids in seeds of Fructus Trichosanthis and the content of 3,29-Dibenzoyl rarounitriol,Karounidiol and the microscopic characteristic constants of stone cells was studied to provide a basis for multidimensional evaluation of the quality of in seeds of Fructus Trichosanthis.5.Through the preparation of different solvent extracts of Fructus Trichosanthes,the antioxidant capacity was compared,and the best antioxidant solvent of Fructus Trichosanthes was selected,and the correlation between the fingerprint of the best solvent and the antioxidant activity was analyzed to provide reference for the rational development and application of Fructus Trichosanthes.Material and method:1.The microscopic characteristic constants of stone cells in Fructus Trichosanthes were determined by microscopic quantitative method.The mathematical models of total triterpenoids content,3,29-Dibenzoyl rarounitriol,Karounidiol content and the microscopiccharacteristic constants of stone cells were established by SPSS statistical software to determine their correlation.2.The optimum preparation process of total triterpenoids in seeds of Fructus Trichosanthis was optimized by Box-behnken response surface methodology.The content of total triterpenoids in in seeds of Fructus Trichosanthis was determined by UV spectrophotometry.3.The content of 3,29-Dibenzoyl rarounitriol in seeds of Fructus Trichosanthis was determined by high performance liquid chromatography with the method of equal elution.Chromatographic column: Agi Lent TC-C18(250 mm *4.6 mm,5 micron);mobile phase:methanol-water(93:7);flow rate: 1 m L/min;column temperature: 30 detection wavelength:230 nm;injection volume: 10 mu L.4.The content of Karounidiol in seeds of Fructus Trichosanthis was determined by HPLC with the method of equal elution.The chromatographic column was Agi Lent TC-C18(250mm *4.6 mm,5 micron);the mobile phase was pure methanol;the flow rate was 1 m L/min;the column temperature was 30 the detection wavelength was 239 nm;and the injection volume was 10 mu L.5.HPLC fingerprints of in seeds of Fructus Trichosanthis were established by gradient elution.Chromatographic column: Agi Lent TC-C18(250 mm *4.6 mm,5 micron);mobile phase A is acetonitrile,mobile phase B is 0.1% phosphoric acid water;flow rate 1 m L / min;column temperature: 30%;detection wavelength: 230 nm;sample volume 10 micro L.6.The water and alcohol extracts of Fructus Trichosanthes were prepared under the condition of 80 ℃ water bath by reflux extraction method,and the dichloromethane extract of Fructus Trichosanthes was prepared by ultrasonic extraction method.Then the antioxidant activities of the three extracts were measured and compared.HPLC was used to establish the fingerprint of the aqueous extract of Fructus Trichosanthis,and the correlation between the fingerprint and the antioxidation was studied.Result:1.Among the 10 habitats,the microscopic characteristic constants of stone cells in seeds of Fructus Trichosanthis differed greatly: the microscopic characteristic constants of Anhui 2,No.10,were the largest,458.8/mg,and that of Xinji,Hebei,No.4,was the smallest,115.3/mg.2.The optimum conditions were as follows: material-liquid ratio 40:1,ultrasonic time 20 min,ultrasonic power 100 W;the content of total triterpenoids in seeds of Fructus Trichosanthis from 10 habitats was different greatly,of which the content of total triterpenoids from No.9(Anhui 1)was the highest,45.71 mg/g,while that from No.7(Shandong)was the lowest.It’s15.22mg/g.3.Among the 10 habitats,the content of 3,29-Dibenzoyl rarounitriol in seeds of Fructus Trichosanthis was the highest in Anguo,1.455mg/g,the lowest in Xinxiang,Henan Province,0.945mg/g,and the content of Karounidiol in seeds of Fructus Trichosanthis was the highest in Dingzhou,0.090mg/g in Shandong Province.The content of 0.02260mg/g was the lowest.A total of 13 chromatographic peaks of in seeds of Fructus Trichosanthis from 10 origins were established by HPLC fingerprints.Among them,13 peaks were the chromatographic peaks of the control substance Trichosanthemum.The similarity of HPLC fingerprints of in seeds of Fructus Trichosanthis from various habitats was greater than 0.941.4.Taking the microscopic characteristic constants of stone cells in seeds of Fructus Trichosanthis from different habitats asdependent variables and the corresponding3,29-Dibenzoyl rarounitriol in seeds of Fructus Trichosanthis as independent variables,the linear regression equation between them is Y=-449.517X+830.56,r=0.657,and the content of total triterpenes in Fructus Trichosanthes from different habitats as dependent variables,and the corresponding 3,29-Dibenzoyl rarounitriol in seeds of Fructus Trichosanthis from different habitats as independent variables.The linear regression equation between them is Y=49.511X-29.031,r=0.753.The contents of total triterpenoids in Fructus Trichosanthes from different habitats were taken as independent variables,and the corresponding contents of3,29-Dibenzoyl rarounitriol and Karounidiol were taken as dependent variables.The regression equation between them was Y=44.018X1+199.914X2-34.970,r=-0.873.there is no correlation between other quantitative models of in seeds of Fructus Trichosanthis.The extraction rates of different solvent extracts from in seeds of Fructus Trichosanthis differ greatly.5.The contents of total triterpenes in different solvent extracts of Trichosanthes are quite different.The contents of total triterpenes in water extract,alcohol extract and dichloromethane extract are 12.72mg/g,18.24mg/g and 30.85mg/g,respectively.The order of antioxidant activity is water extract > alcohol extract > dichloromethane extract.The VIP values of DPPH and ABTS free radicals scavenged by the eight peaks of 11,9,6,12,3,4,14 and 10 in the aqueous extract of Fructus Trichosanthes are all greater than 1,which indicates that these peaks play an important role in antioxidation of the aqueous extract of Fructus Trichosanthes.Conclusion:1.The microscopic characteristic constants of stone cells of in seeds of Fructus Trichosanthis from 10 different habitats are different,which can be used as a basis for differentiating the quality of in seeds of Fructus Trichosanthis from different habitats.The method is stable and reliable.It provides a basis for the qualitative identification of Fructus Trichosanthes from different producing areas and the formulation of microscopic quantitative standards for stone cells.It also lays a foundation for the correlation analysis between microscopic characteristic constants and chemical constituents.2.Box-behnken Response Surface Methodology was used to optimize the determination of total triterpenoids in seeds of Fructus Trichosanthis and could be used to determine the total triterpenoids in seeds of Fructus Trichosanthis.The contents of total three terpenoids in seeds of Fructus Trichosanthis from different habitats are quite different,The content of total three terpenoids can be used to determine the quality standard content of in seeds of Fructus Trichosanthis.3.Karounidiol and 3,29-Dibenzoyl rarounitriol as the exclusive components of in seeds of Fructus Trichosanthis,both can be used as two important indicators in quantitative quality control of in seeds of Fructus Trichosanthis.The HPLC fingerprint of in seeds of Fructus Trichosanthis can provide a scientific basis for qualitative identification of in seeds of Fructus Trichosanthis.4.The microscopic characteristic constants of stone cells in seeds of Fructus Trichosanthis were negatively correlated with the content of 3,29-Dibenzoyl rarounitriolin seeds of Fructus Trichosanthis.The content of total triterpenes in Fructus Trichosanthes was positively correlated with the content of 3,29-Dibenzoyl rarounitriol in seeds of Fructus Trichosanthis.The content of total triterpenes in Fructus Trichosanthes was positively correlated with the content of 3,29-Dibenzoyl rarounitriol and Karounidiol in seeds of Fructus Trichosanthis.The method is stable and feasible.It provides a new method and a new means for the quality determination of Trichosanthes fructus.It also provides a basis for formulating the quality standard of Fructus Trichosanthis.5.The water extract of in seeds of Fructus Trichosanthis has the strongest antioxidant activity.The main antioxidant component of in seeds of Fructus Trichosanthis is not total triterpenoids.