CN107550964A - Response phase method optimizes Ageratum conyzoides extracting flavonoids technique and its application - Google Patents
Response phase method optimizes Ageratum conyzoides extracting flavonoids technique and its application Download PDFInfo
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- CN107550964A CN107550964A CN201711003886.5A CN201711003886A CN107550964A CN 107550964 A CN107550964 A CN 107550964A CN 201711003886 A CN201711003886 A CN 201711003886A CN 107550964 A CN107550964 A CN 107550964A
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- 150000002215 flavonoids Chemical class 0.000 title claims abstract description 31
- 235000017173 flavonoids Nutrition 0.000 title claims abstract description 31
- 238000000605 extraction Methods 0.000 claims abstract description 103
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- 235000011949 flavones Nutrition 0.000 claims abstract description 84
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Abstract
The invention discloses a kind of response phase method optimization Ageratum conyzoides extracting flavonoids technique and its application, according to Box Behnken experimental design principles, using the Design Expert Software Design Optimizations extraction concentration of alcohol of Ageratum conyzoides flavones, extraction time, solid-liquid ratio, extraction time process conditions, Ageratum conyzoides flavones is extracted using ultrasonic assistant, comprised the following steps that：Ageratum conyzoides is dried, is ground into powder, Ageratum conyzoides powder is put into extractor to mix with ethanol solution and carries out ultrasonic wave extraction, extract solution is placed in Rotary Evaporators and is evaporated under reduced pressure, and evaporation of solvent of divulging information, water bath method is recycled, obtains Ageratum conyzoides flavones product.The technique extraction yield of the present invention is high, and the Ageratum conyzoides flavones oxidation resistance extracted is strong, has stronger application effect in anti-oxidation medicine and food antioxidant.
Description
【Technical field】
The invention belongs to medicinal plant active ingredient and food additives extractive technique field, and in particular to a kind of response surface
Method optimizes Ageratum conyzoides extracting flavonoids technique and its application.
【Background technology】
Ageratum conyzoides (Ageratum ConyzoidesL.), alias Ageratum conyzoides, halfspreading ironweed herb, Pemisetum flaccidum Griseb, anti-inflammatory be careless, smelly black earth grass
Deng being composite family ageratum annual herb plant.Introduce a fine variety incoming China and be now distributed widely in each provinces and regions on the south the Changjiang river.It is incorporated in
South China each province Chinese medicine provincial standard
Ageratum conyzoides have it is clearing heat and detoxicating, hemostasis, analgesia the effect of, be countries in the world Traditional Folk medicinal plant, Asia,
There is traditional medicinal history on the ground such as Africa, South America, be mainly used in treat headache, expiratory dyspnea, skin disease, vaginitis, pneumonia,
Relieving asthma, stop spasm, hemostasis, antipruritic, desinsection etc..And southern region of China it is among the people be mainly used in treating cat fever, throat swell
Bitterly, the disease such as ulcer boil, traumatism and bleeding；Clinically it is mainly used in the diseases such as pelvic infecton, tonsillitis, pharyngitis, has preferable
Curative effect.
Modern pharmacology research shows Ageratum conyzoides in anti-inflammatory, analgesic, antipyretic, radioresistance, stomach protective effect etc. wrinkled giant hyssop
Ji has very high bioactivity.In order to disclose the effective component of Ageratum conyzoides pharmacological action, chemistry of the scholars to Ageratum conyzoides
Composition is also studied, and is as a result found, Ageratum conyzoides chemical composition is various, and that reports at present has volatile oil, flavones, terpene, steroid
Alcohol, alkaloid and amino acid etc..It is main wherein in Ageratum conyzoides volatile oil to contain monoterpene and sesquiterpenoids, and the place of production is different,
Contained volatile oil component and content has certain difference.Flavonoids it is separated go out more than 20 kinds of flavone compound, wherein 14 kinds
For polymethoxyflavone class compound, and domestic scholars fly to wait research discovery recklessly, and Ageratum conyzoides Flavonoid substances are main to Citrus Orchard
Pathogen has very strong inhibitory action, and the bacteriostatic activity for the flavones molecule that can be especially accumulated in Citrus Orchard soil exceedes business
Industry bactericide.Alkaloid compound mainly has 4 kinds.Triterpenes and phytosterin compound also have 4 kinds.In addition other compositions also have sesame
Numb element (Z-sesamin), amidoacetic acid ester (aurantiamide acetate), fumaric acid (fumaric acid), coffee
Sour (caffeic acid), aliphatic acid, amino acid and vitamin A and C etc..
Find that it includes abundant flavonoids and terpene compound from Ageratum conyzoides chemical composition analysis document, this is exactly
The maximally effective composition of Chinese medicine anti-oxidation, but domestic and foreign literature is showed no Ageratum conyzoides extracting flavonoids technique and its anti-oxidant at present
The report of Effect study.Particularly in China, Ageratum conyzoides is taken as weeds processing always, and it is studied and also only rests on chemurgy
In terms of sense effect, but seldom it is related to for Ageratum conyzoides drug efficacy study.
Ageratum conyzoides has substantial amounts of wild resource in Guangxi District, is typically regarded as weeds processing at this stage.In order to abundant
Excavate Ageratum conyzoides medical value, study Ageratum conyzoides flavones extraction and its oxidation resistance analysis, be Ageratum conyzoides exploitation into
Theoretical foundation is provided for natural.
【The content of the invention】
The invention provides a kind of response phase method optimization Ageratum conyzoides extracting flavonoids technique and its application, concrete technical scheme is such as
Under：
A kind of response phase method optimizes Ageratum conyzoides extracting flavonoids technique, according to Box-Behnken experimental design principles, utilizes
The Design-Expert Software Design Optimizations extraction concentration of alcohol of Ageratum conyzoides flavones, extraction time, solid-liquid ratio, extraction time
Process conditions, Ageratum conyzoides flavones is extracted using ultrasonic assistant, comprised the following steps that：
S1：Ageratum conyzoides complete stool is dried, 20-60 mesh sieves are crossed after crushing, it is standby to weigh Ageratum conyzoides powder 2.0g；
S2：Ageratum conyzoides powder is put into extractor, adds 30-50% ethanol solution, solid-liquid ratio 1：(20-40),
Cold soaking 24h；
S3：Ageratum conyzoides powder after cold soaking is subjected to ultrasonic wave extraction, extraction time 20-40min, extraction time 1-
3 times, filtering, merge extract solution；
S4：Extract solution is placed in Rotary Evaporators and is evaporated under reduced pressure, recycling design, the evaporation of solvent in fume hood, then
Using water bath method, Ageratum conyzoides flavones product is obtained.
As optimal preferred, described response phase method optimization Ageratum conyzoides extracting flavonoids technique, tested according to Box-Behnken
Design principle, utilize the concentration of alcohol of Design-Expert Software Design Optimizations extraction Ageratum conyzoides flavones, extraction time, feed liquid
Than the process conditions of, extraction time, Ageratum conyzoides flavones is extracted using ultrasonic assistant, comprised the following steps that：
S1：Ageratum conyzoides complete stool is dried, 60 mesh sieves are crossed after crushing, it is standby to weigh Ageratum conyzoides powder 2.0g；
S2：Ageratum conyzoides powder is put into extractor, adds 48% ethanol solution, solid-liquid ratio 1：40, cold soaking 24h；
S3：Ageratum conyzoides powder after cold soaking is subjected to ultrasonic wave extraction, extraction time 24min, extraction time 3 times, mistake
Filter, merge extract solution；
S4：Extract solution is placed in Rotary Evaporators and is evaporated under reduced pressure, recycling design, the evaporation of solvent in fume hood, then
Using water bath method, Ageratum conyzoides flavones product is obtained.
Preferably, Ageratum conyzoides powder described in step S2 should shake up once during cold soaking every 2h.
Preferably, the feed temperature control described in step S3 during ultrasonic wave extraction is 50-60 DEG C.
Preferably, Rotary Evaporators rotating speed described in step S4 is 120r/min, and pressure 0.06-0.08MPa, temperature
For 70-75 DEG C.
It is evaporated preferably, water bath method described in step S4 is 70 DEG C of constant temperature water baths.
It is a further object of the present invention to provide Ageratum conyzoides flavones made from described extraction process to prepare anti-oxidation medicine
With the application of food antioxidant.
Described anti-oxidation medicine and the oxidation resistance of food antioxidant increases with the increase of Ageratum conyzoides flavones content
By force.
The anti-oxidation medicine and food antioxidant possess the Scavenging activity to free radical.
In one of the embodiments, the free radical includes DPPH free radicals and ABTS+Free radical.
The invention has the advantages that：
(1) pharmacological action and clinical practice of Ageratum conyzoides, and the research to Ageratum conyzoides chemical composition of volatile oil are a lot, but
Research on Ageratum conyzoides oxidation resistance, which but has no, to be had been reported that.Invention herein utilizes 30-50% ethanol using Ageratum conyzoides as raw material
Solution is simultaneously aided with ultrasonic wave Ageratum conyzoides flavones is extracted, using DPPH methods, ABTS+Method and phosphomolybdate method evaluation wrinkled giant hyssop
The oxidation resistance of Ji chromocor extract, scientific basis is provided for the Ageratum conyzoides utilization of resources；
(2) response phase method optimization Ageratum conyzoides extracting flavonoids technique provided by the present invention and its application, according to Box-
Behnken experimental design principles, extract the concentration of alcohol of Ageratum conyzoides flavones using Design-Expert Software Design Optimizations, carry
Time, solid-liquid ratio, the process conditions of extraction time are taken, Ageratum conyzoides flavones are extracted using ultrasonic assistant, purifying difficulty is low, yield
Height, cost is low, removes free radical and oxidation resistance is strong, have stronger application in anti-oxidation medicine and food antioxidant
Effect.
【Brief description of the drawings】
Influence figure of Fig. 1 time gradients to extracting flavonoids；
Influence figure of Fig. 2 concentration of alcohol to extracting flavonoids；
Influence figure of Fig. 3 solid-liquid ratios to extracting flavonoids；
Fig. 4 Y-f (X1,X2)) response surface and contour map；
Fig. 5 Y=f (X1,X3) response surface and contour map；
Fig. 6 Y=f (X1,X4) response surface and contour map；
Fig. 7 Y=f (X2,X3)Response surface and contour map；
Fig. 8 Y=f (X2,X4) response surface and contour map；
Fig. 9 Y=f (X3,X4) response surface and contour map；
Figure 10 Ageratum conyzoides flavones TAC figures；
Scavenging activity figure of Figure 11 Ageratum conyzoides flavones to DPPH free radicals；
Figure 12 Ageratum conyzoides flavones is to ABTS+The Scavenging activity figure of free radical.
【Embodiment】
It is specifically described with reference to specific embodiment.
Embodiment 1：
A kind of response phase method optimizes Ageratum conyzoides extracting flavonoids technique, according to Box-Behnken experimental design principles, utilizes
The Design-Expert Software Design Optimizations extraction concentration of alcohol of Ageratum conyzoides flavones, extraction time, solid-liquid ratio, extraction time
Process conditions, Ageratum conyzoides flavones is extracted using ultrasonic assistant, comprised the following steps that：
S1：By Ageratum conyzoides complete stool under conditions of cool place natural drying, after crushing cross 60 mesh sieves, weigh Ageratum conyzoides powder
It is 2.0g standby；
S2：Ageratum conyzoides powder is put into extractor, adds 48% ethanol solution, solid-liquid ratio 1：40, cold soaking 24h,
Shaken up once every 2h；
S3：Ageratum conyzoides powder after cold soaking is subjected to ultrasonic wave extraction, feed temperature control when taking is 50 DEG C, during extraction
Between be 24min, extraction time is 3 times, filtering, merges extract solution；
S4：It is 120r/min that extract solution is placed in into rotating speed, and pressure 0.06MPa, temperature is in 70 DEG C of Rotary Evaporators
It is evaporated under reduced pressure, recycling design, while the evaporation of solvent in fume hood, recycle 70 DEG C of constant temperature water baths to be evaporated, obtain Ageratum conyzoides
Flavones product 396mg.
Embodiment 2：
A kind of response phase method optimizes Ageratum conyzoides extracting flavonoids technique, according to Box-Behnken experimental design principles, utilizes
The Design-Expert Software Design Optimizations extraction concentration of alcohol of Ageratum conyzoides flavones, extraction time, solid-liquid ratio, extraction time
Process conditions, Ageratum conyzoides flavones is extracted using ultrasonic assistant, comprised the following steps that：
S1：By Ageratum conyzoides complete stool under conditions of cool place natural drying, after crushing cross 60 mesh sieves, weigh Ageratum conyzoides powder
It is 2.0g standby；
S2：Ageratum conyzoides powder is put into extractor, adds 48% ethanol solution, solid-liquid ratio 1：40, cold soaking 24h,
Shaken up once every 2h；
S3：Ageratum conyzoides powder after cold soaking is subjected to ultrasonic wave extraction, feed temperature control when taking is 55 DEG C, during extraction
Between be 24min, extraction time is 3 times, filtering, merges extract solution；
S4：It is 120r/min that extract solution is placed in into rotating speed, and pressure 0.07MPa, temperature is in 72 DEG C of Rotary Evaporators
It is evaporated under reduced pressure, recycling design, while the evaporation of solvent in fume hood, recycle 70 DEG C of constant temperature water baths to be evaporated, obtain Ageratum conyzoides
Flavones product 407mg.
Embodiment 3：
A kind of response phase method optimizes Ageratum conyzoides extracting flavonoids technique, according to Box-Behnken experimental design principles, utilizes
The Design-Expert Software Design Optimizations extraction concentration of alcohol of Ageratum conyzoides flavones, extraction time, solid-liquid ratio, extraction time
Process conditions, Ageratum conyzoides flavones is extracted using ultrasonic assistant, comprised the following steps that：
S1：By Ageratum conyzoides complete stool under conditions of cool place natural drying, after crushing cross 60 mesh sieves, weigh Ageratum conyzoides powder
It is 2.0g standby；
S2：Ageratum conyzoides powder is put into extractor, adds 48% ethanol solution, solid-liquid ratio 1：40, cold soaking 24h,
Shaken up once every 2h；
S3：Ageratum conyzoides powder after cold soaking is subjected to ultrasonic wave extraction, feed temperature control when taking is 60 DEG C, during extraction
Between be 24min, extraction time is 3 times, filtering, merges extract solution；
S4：It is 120r/min that extract solution is placed in into rotating speed, and pressure 0.08MPa, temperature is in 75 DEG C of Rotary Evaporators
It is evaporated under reduced pressure, recycling design, while the evaporation of solvent in fume hood, recycle 70 DEG C of constant temperature water baths to be evaporated, obtain Ageratum conyzoides
Flavones product 383mg.
Embodiment 4：
A kind of response phase method optimizes Ageratum conyzoides extracting flavonoids technique, according to Box-Behnken experimental design principles, utilizes
The Design-Expert Software Design Optimizations extraction concentration of alcohol of Ageratum conyzoides flavones, extraction time, solid-liquid ratio, extraction time
Process conditions, Ageratum conyzoides flavones is extracted using ultrasonic assistant, comprised the following steps that：
S1：By Ageratum conyzoides complete stool under conditions of cool place natural drying, after crushing cross 60 mesh sieves, weigh Ageratum conyzoides powder
It is 2.0g standby；
S2：Ageratum conyzoides powder is put into extractor, adds 48% ethanol solution, solid-liquid ratio 1：40, cold soaking 24h,
Shaken up once every 2h；
S3：Ageratum conyzoides powder after cold soaking is subjected to ultrasonic wave extraction, feed temperature control when taking is 58 DEG C, during extraction
Between be 24min, extraction time is 3 times, filtering, merges extract solution；
S4：It is 120r/min that extract solution is placed in into rotating speed, and pressure 0.078MPa, temperature is in 75 DEG C of Rotary Evaporators
It is evaporated under reduced pressure, recycling design, while the evaporation of solvent in fume hood, recycle 70 DEG C of constant temperature water baths to be evaporated, obtain Ageratum conyzoides
Flavones product 415mg.
Embodiment 5：In order to inquire into influence of the single factor test extraction to Ageratum conyzoides flavones content, while carry out Box-Behnken
Experimental design and response surface analysis, and the oxidation resistance of Ageratum conyzoides flavones is evaluated, make following experiment.
First, single factor test extracts the influence to Ageratum conyzoides flavones content
1. influence of the time to flavones content
Using the Ageratum conyzoides powder 2.0g of 60 mesh sieves excessively as raw material, by 1:40 liquid ratio adds 40% ethanol extractant, if
Fixed different extraction time：20、30、40、50、60、90min.Flavones content is determined after extraction, as a result as shown in Figure 1.
Before extraction time is 30min, flavones content increases with the increase of extraction time in extract solution,
Content highest during 30min, flavones content 46.931mg, hereafter yield declined.It can make because extraction time extends
Flavones is destructurized or oxidation occurs, polymerize and loses.Therefore, continue to examine using extraction time 30min as follow-up test
Examine.
2. influence of the volume fraction of ethanol to flavones content
Respectively with 30,40,50,60,70,80,95% concentration of alcohol, 30min is extracted.Flavones content is determined after extraction,
As a result it is as shown in Figure 2.
From Figure 2 it can be seen that having content peak 65.485mg when concentration of alcohol is 40%, hereafter contain with the rise of concentration
Amount have dropped on the contrary, so the Ageratum conyzoides powder for crossing 60 mesh sieves is raw material, by 1:40 solid-liquid ratio, which adds, sets different extractions
Concentration of alcohol.For cost and the difficulty of reduction subsequent concentration purifying, it is 40% to select concentration of alcohol.
3. influence of the solid-liquid ratio to flavones content
Using the Ageratum conyzoides powder 2.0g for crossing 60 mesh sieves as raw material, the different solid-liquid ratios of setting are pressed with 40% ethanol：1:10、
1:15、1:20、1:30、1:40、1:50 extraction 30min, determine flavones content, as a result as shown in Figure 3 after extraction.
As seen from Figure 3, solid-liquid ratio 1:Before 30, flavones content raises with the increase of solid-liquid ratio, and 1:When 30
There is content peak 75.857mg；And 1:The trend unobvious that flavones content rises after 30.So with reference to other single factor tests
Influence, and for cost and the difficulty of reduction subsequent concentration purifying, it is 1 to select solid-liquid ratio:30.
4. influence of the extraction time to flavones content
Using the Ageratum conyzoides powder 2.0g for crossing 60 mesh sieves as raw material, solid-liquid ratio 1 is fixed:40, concentration of alcohol 40%, during extraction
Between be 30min, investigate influence of the extraction time to flavones content, as a result as shown in table 1, after 2 extractions, flavones content increases
It is significantly smaller, so for cost and the difficulty of reduction subsequent concentration purifying, selection and withdrawal number is 3 times.
Table 1：Influence of the extraction time to extracting flavonoids
| Extraction time/n | 1 | 2 | 3 |
| Flavones content/mg | 43.110 | 84.571 | 87.118 |
2nd, Box-Behnken experimental designs and response surface analysis
The process conditions of flavones in Ageratum conyzoides are extracted for optimization, according to Box-Behnken experimental design principles, in single factor test
The optimization of extraction conditions is carried out in experimental basis using the horizontal response surface design analysis method of four factor three：Precision weighs the 2.0g leaves of pulse plants
Fragrant Ji sample chooses concentration of alcohol (X in triangular flask1), extraction time (X2), solid-liquid ratio (X3), extraction time (X4) four it is right
The factor that Ageratum conyzoides effective component extraction rate has a great influence, to extraction process carry out response surface analysis, its specific testing program and
It the results are shown in Table 2,3.
Table 2：Response surface experiments design factor water-glass
Table 3：Response surface experiments design and result
1. the foundation and its significance test of model
According to the data of table 3, multiple regression Fitting Analysis is carried out with the softwares of Design Expert 7.0, it is auxiliary to obtain ultrasound
The predicted value (Y) of extraction yogimbine extracted amount is helped to independent variable volume fraction of ethanol (X1), extraction time (X2), solid-liquid ratio (X3)、
Extraction time (X4) secondary multiple regression equation it is as follows：
Y=155.649-4.0597X1-0.96123·X2-4.34823·X3-11.2577·X4+0.0556·X1·
X2+0.01542·X1·X3+0.60842·X1·X4-0.0429·X2·X3+0.11893·X2·X4+0.0133·X1 2+
0.1006·X3 2-2.5156·X4 2
Regression model equation coefficient, variance analysis and the result of the significance test of factor according to the result of the test of table 3
It is shown in Table 4.
Table 4：Regression model significance test
Note：* differences are extremely notable, p<0.01；Significant difference, p<0.05
From the results of analysis of variance table 4, the F=3.24, p=of the regression equation mathematical modeling of Ageratum conyzoides extracting flavonoids
0.0177(p<0.05), show that the fitting of the regression model equation reaches the level of signifiance.The coefficient of determination R of regression model equation2=
0.9013, adjust coefficient R2 Adj=0.8018, this shows that the fitting degree of the model equation and test data is high, test group
Error it is small, so model equation be fully available for Ageratum conyzoides flavones ultrasound assisted extraction result analysis and prediction instruct.
Simultaneously by the results of analysis of variance table 4 it can also be seen that, in first order, solid-liquid ratio and extraction time are to extracting flavonoids amount
With significant impact (p<0.01 or p<0.05), other factorses influence not notable；In quadratic term, solid-liquid ratio (C2) reach non-
Normal significance (p<0.01)；In interaction item, not significantly (p is influenceed>0.05) reciprocation between each factor, is prompted not
Significantly.From F values, influence size of each factor to extracting flavonoids amount is followed successively by C (solid-liquid ratio) ＞ D (extraction time) ＞ A (second
Determining alcohol) ＞ B (extraction time).
2. response surface design analysis and optimization
Secondary polynomial fitting, which is carried out, using the softwares of Design Expert 7.0 obtains the response surface song of quadratic regression equation
Line, in the case where fixing other two factor levels values and being zero, another two factors reciprocation is investigated to Ageratum conyzoides extracting flavonoids amount
(Y) influence, gained response surface is as shown in figures 4-9.If the gradient of a response surface design is shallower, show with treatment conditions
Change, response value changes it is smaller；And if the gradient of a response surface design is more precipitous, show the change with treatment conditions,
Response changes greatly.
From Fig. 5, Fig. 7 and Fig. 9, the gradient of response surface is steeper, shows solid-liquid ratio and concentration of alcohol, solid-liquid ratio and extraction
The reciprocation of time, solid-liquid ratio and extraction time has considerable influence to flavones yield；It may determine that from the density of contour
Go out, influence of the solid-liquid ratio to response will be big than concentration of alcohol, extraction time and extraction time；Can from Fig. 4, Fig. 6 and Fig. 8
Go out, the gradient of response surface is shallower, and concentration of alcohol and extraction time, the reciprocation of concentration of alcohol and extraction time obtain to flavones
Rate influences smaller, may determine that from the density of contour, influence of the extraction time to response is more than concentration of alcohol, and second
Influence of the determining alcohol to response is more than extraction time.In summary, influence of the solid-liquid ratio to flavones yield is maximum, extraction time
Take second place, the influence of extraction time is minimum；And with the rise of solid-liquid ratio and extraction time, flavones content increase.
3. the determination of optimum condition and the checking of model
Using the mathematical modeling established, optimization optimal conditions is that concentration of alcohol is 48.04% in scope of experiment, is carried
It is 23.9min to take the time, solid-liquid ratio 1:40；Extraction time is 3；On this condition, ultrasonic wave added is from Ageratum conyzoides extracting flavonoids
Measure as 427.03mg.In view of practical operation condition, it is 48%, extraction time 24min that condition is modified into concentration of alcohol, is expected
Liquor ratio is 1:40；Extraction time is 3.Proving test is carried out using above-mentioned condition, carries out 3 parallel tests, experimental result table
Bright, it is 415mg that can obtain flavones average value with 2g Ageratum conyzoides, and compared with theoretical expectation values, its relative error is about 2.81%, explanation
Optimum results are reliable.
3rd, the oxidation resistance evaluation experimental of Ageratum conyzoides flavones.
1. the measure of Ageratum conyzoides flavones TAC
The optimal extraction conditions obtained using response surface analysis extract Ageratum conyzoides flavones, and obtained Ageratum conyzoides flavones is produced
Product, it is made into following serial solution：5mg/ml、10mg/ml、15mg/ml、20mg/ml、25mg/ml、30mg/ml、35mg/ml
Deng, by above-mentioned serial solution do Ageratum conyzoides flavones oxidation resistance test, as a result see Figure 10.
As can be seen from Figure 10, Ageratum conyzoides flavones has very strong TAC, and TAC is with wrinkled giant hyssop
The increase of Ji flavones concentration and strengthen.
2. Ageratum conyzoides flavones is to the clearance rate of DPPH free radicals
The optimal extraction conditions obtained using response surface analysis extract Ageratum conyzoides flavones, and obtained Ageratum conyzoides flavones is produced
Product are made into following serial solution：5mg/ml, 10mg/ml, 15mg/ml, 20mg/ml, 25mg/ml, 30mg/ml, 35mg/ml,
Above-mentioned serial solution is done into the experiment of DPPH radical scavenging activities, as a result sees Figure 11.
As can be seen from Figure 11, Ageratum conyzoides flavones has a very strong DPPH Scavenging activities, half elimination ratio 22.78mg/ml, and
And Scavenging activity (clearance rate) strengthens with the increase of Ageratum conyzoides flavones concentration.
3. Ageratum conyzoides flavones is to ABTS+The clearance rate of free radical
The optimal extraction conditions obtained using response surface analysis extract Ageratum conyzoides flavones, and obtained Ageratum conyzoides flavones is produced
Product are made into following serial solution：5mg/ml, 10mg/ml, 15mg/ml, 20mg/ml, 25mg/ml, 30mg/ml, 35mg/ml,
Above-mentioned serial solution is done into ABTS+Radical scavenging activity is tested, and as a result sees Figure 12.
As can be seen from Figure 12, Ageratum conyzoides flavones has stronger ABTS+Radical scavenging activity, and Scavenging activity is (clear
Except rate) strengthen with the increase of Ageratum conyzoides flavones concentration.
Above content is to combine specific preferred embodiment further description made for the present invention, it is impossible to is assert
The specific implementation of the present invention is confined to these explanations, for person of an ordinary skill in the technical field, is not departing from
On the premise of present inventive concept, some simple deduction or replace can also be made, should all be considered as belonging to the present invention by being submitted
Claims determine scope of patent protection.
Claims (10)
1. a kind of response phase method optimizes Ageratum conyzoides extracting flavonoids technique, it is characterised in that：It is former according to Box-Behnken experimental designs
Reason, utilize the concentration of alcohol of Design-Expert Software Design Optimizations extraction Ageratum conyzoides flavones, extraction time, solid-liquid ratio, extraction
The process conditions of number, Ageratum conyzoides flavones is extracted using ultrasonic assistant, comprised the following steps that：
S1：Ageratum conyzoides complete stool is dried, 20-60 mesh sieves are crossed after crushing, it is standby to weigh Ageratum conyzoides powder 2.0g；
S2：Ageratum conyzoides powder is put into extractor, adds 30-50% ethanol solution, solid-liquid ratio 1：(20-40), cold soaking
24h；
S3：Ageratum conyzoides powder after cold soaking is subjected to ultrasonic wave extraction, extraction time 20-40min, extraction time is 1-3 times,
Filtering, merge extract solution；
S4：Extract solution is placed in Rotary Evaporators and is evaporated under reduced pressure, recycling design, the evaporation of solvent in fume hood, is recycled
Water bath method, obtain Ageratum conyzoides flavones product.
2. response phase method according to claim 1 optimizes Ageratum conyzoides extracting flavonoids technique, it is characterised in that：According to Box-
Behnken experimental design principles, extract the concentration of alcohol of Ageratum conyzoides flavones using Design-Expert Software Design Optimizations, carry
Time, solid-liquid ratio, the process conditions of extraction time are taken, Ageratum conyzoides flavones, including following technique step are extracted using ultrasonic assistant
Suddenly：
S1：Ageratum conyzoides complete stool is dried, 60 mesh sieves are crossed after crushing, it is standby to weigh Ageratum conyzoides powder 2.0g；
S2：Ageratum conyzoides powder is put into extractor, adds 48% ethanol solution, solid-liquid ratio 1：40, cold soaking 24h；
S3：Ageratum conyzoides powder after cold soaking is subjected to ultrasonic wave extraction, extraction time 24min, extraction time 3 times, filters, closes
And extract solution；
S4：Extract solution is placed in Rotary Evaporators and is evaporated under reduced pressure, recycling design, the evaporation of solvent in fume hood, is recycled
Water bath method, obtain Ageratum conyzoides flavones product.
3. response phase method according to claim 1 or 2 optimizes Ageratum conyzoides extracting flavonoids technique, it is characterised in that：Step S2
Described in Ageratum conyzoides powder during cold soaking, should be shaken up once every 2h.
4. response phase method according to claim 1 or 2 optimizes Ageratum conyzoides extracting flavonoids technique, it is characterised in that：Step S3
Described in ultrasonic wave extraction when feed temperature control be 50-60 DEG C.
5. response phase method according to claim 1 or 2 optimizes Ageratum conyzoides extracting flavonoids technique, it is characterised in that：Step S4
Described in Rotary Evaporators rotating speed be 120r/min, pressure 0.06-0.08MPa, temperature is 70-75 DEG C.
6. response phase method according to claim 1 or 2 optimizes Ageratum conyzoides extracting flavonoids technique, it is characterised in that：Step S4
Described in water bath method be 70 DEG C of constant temperature water baths be evaporated.
7. Ageratum conyzoides flavones made from the extraction process according to claim any one of 1-6 is preparing anti-oxidation medicine and food
The application of product antioxidant.
8. application according to claim 7, it is characterised in that：The anti-oxidation medicine and food antioxidant it is anti-oxidant
Ability strengthens with the increase of Ageratum conyzoides flavones content made from the extraction process described in claim any one of 1-6.
9. application according to claim 7, it is characterised in that：The anti-oxidation medicine and food antioxidant possess to certainly
By the Scavenging activity of base.
10. application according to claim 9, it is characterised in that：The free radical includes DPPH free radicals and ABTS+From
By base.
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