Study On The Extraction Process Of Sinigrin In Thlaspiarvense And Its Antioxidant And Bacteriostasis Effects In Vitro

Thlaspi arvense L.is an annual herbaceous plant of the genus Thlaspi in the cruciferae family,known as defeated sauce grass,curb bluecabbage,and ploughshares.It is widely distributed throughout the country,and grows intensively on ditches,mountain roadsides and around villages T.arvense contains volatile oils,flavonoids,glucosinolates and other compounds,as a representative compound of glucosinolates,with anti-lipogenic effect,with helpinginternal organs,wick off perspiration and cooling out,mild appetizing,enhance appetite,anti-inflammatory,antioxidant activity and other effects.In this experiment,the extraction rate of sinigrin in T.arvense seeds was improved,a high performance liquid chromatography detection method for sinigrin was established,on the basis of determining the optimal enzyme and univariate experiments,The process of enzymatic hydrolysis-assisted ultrasound extraction method was optimized by response surface method,the antioxidant activity and bacteriostatic activity of sinigrin were studied.It provides data support for the production practice of sinigrin in T.arvense,and lays a theoretical foundation for the in-depth development of T.arvense.The research content and results of this experiment are as follows:1.In this paper,a high performance liquid chromatography detect method for the determination of the target product sinigrin was established with the goal of extracting sinigrin from T.arvense seeds:Mobile phases A and B are methanol and 0.5%aqueous phosphoric acid（A:B is 5:95）.Detection wavelength:215 nm,column temperature:25°C;Flow rate:1m L·min^-1;Injection volume:10μL;Elute with an isocratic gradient for 10 min.Five enzymes（acid protease,neutral protease,alkaline protease,cellulase,pectinase）were screened to obtain the optimal enzyme for subsequent experiments.The optimal extraction conditions were determined by the pre-experiment of ultrasonic extraction method,and the extraction experiment was carried out by this method combined with enzymatic hydrolysis.The process optimization of the extraction of sinigrin from T.arvense seeds by response surface method and the regression equation were established,and the optimal process conditions for extracting sinigrin were as follows:material-liquid ratio 1:15,ultrasonic power 360 W,ultrasonic time25min,enzyme addition amount of 2.90%,enzymatic hydrolysis time of 17.90 min,enzymatic hydrolysis temperature of 49.05°C,at this time,the predicted content of sinigrin content was2.687 mg·g^-1,and the actual content of sinigrin was 2.679 mg·g^-1,The difference between and the predicted value of the response surface was not significant（P>0.05）.Compared with enzymatic hydrolysis-assisted ultrasound extraction,ultrasonic extraction and Soxhlet extraction,the content of sinigrin from T.arvense seeds obtained by enzymatic hydrolysis-assisted ultrasonic extraction method was significantly higher than that of the other two extraction methods.2.The water extract of sinigrin was obtained by ultrasonic extraction method and enzymatic hydrolysis-assisted ultrasonic extraction method,and different concentrations of sinigrin samples were prepared for antioxidant test analysis.The results showed that the sinigrin aqueous extract obtained by the two extraction methods had certain antioxidant capacity and certain reducing ability to iron ions,and the antioxidant activity of enzymatic hydrolysis-assisted ultrasonic extraction method was higher than that of the target product of ultrasonic extraction.With the increase of the concentration of the target product,the antioxidant activity is also enhanced,but it is weaker than the antioxidant activity of ascorbic acid.3.The bacteriostatic activity of sinigrin aqueous extract was detected by agar filter paper diffusion method,and the antibacterial activity of sinigrin aqueous extract was judged by the size of the inhibition circle diameter.The results showed that the water extract of sinigrin had a certain inhibitory effect on Escherichia coli,Staphylococcus aureus,Bacillus subtilis,Candida albicans and Acinetobacter baumannii,among which the inhibitory effect was the best against Acinetobacter baumannii and the weakest effect on Escherichia coli.The minimum inhibitory concentrations（MIC）of the five tested bacteria were determined by microdilution method at25,12.5,6.25,25,and 6.25 mg·m L^-1,and the minimum bactericidal concentrations（MBC）were 50,50,12.5,50,and 12.5 mg·m L^-1,respectively.In summary,the enzymatic hydrolysis-assisted ultrasound extraction method is conducive to the extraction of the active ingredients of sinigrin,which is convenient for the precipitation of sinigrin,and is feasible in improving the yield of sinigrin.Sinigrin water extract has certain antioxidant activity and bacteriostatic activity,which can be used to produce health food and be used in the pharmaceutical industry,providing a wide range of ways for the utilization of T.arvense seed resources.Sinigrin water extract has certain antioxidant activity and bacteriostatic activity,which can be used to produce health food and be used in the pharmaceutical industry,providing a wide range of ways for the utilization of T.arvense seed resources.