CN103494797B - Composition of peony seed total stilbene compounds as well as extraction method and application thereof - Google Patents
Composition of peony seed total stilbene compounds as well as extraction method and application thereof Download PDFInfo
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Abstract
The invention relates to a composition of peony seed total stilbene compounds as well as an extraction method and application thereof. The extraction method comprises the following steps: carrying out reflux extraction on peony seed skins by using ethanol, and filtering and concentrating to obtain an extract; then, dissolving the extract and mixing with an absorber; then, sequentially extracting the mixture by solvents; and finally, refining by using acetone and active carbon and concentrating till no solvents exist, thus obtaining the composition of the total stilbene compounds. The six compounds are respectively peony alcohol A, peony alcohol, peony alcohol C, cis-epsilon viniferin, trans-epsilon viniferin and common jointfir stem H. Verified by pharmacological experiments, the composition of the peony total stilbene compounds has good anti-gram positive and negative bacteria activity, can be used as an active component of anti-bacterial medicines and can be used independently or together with appropriate excipients to serve as a food additive for preservation of foods or can be used for preparing anti-bacterial medicines.
Description
Technical field
The present invention relates generally to the compositions of the total stilbene compound of a kind of seed of Flos Moutan and extracting method thereof and application.
Background technology
Stilbene compound (stilbeniods) refers to have the monomer of stilbene skeleton and the general name of polymer thereof.Oligomeric stilbene class belongs to phenolic acid compound, and this compounds has biological activity widely, as antibacterium and antifungal activity, and anti-inflammatory activity, antioxidant activity, anti-tumor activity, anti-HIV activity and liver-protecting activity etc.[Li little Mei, Li Na, Huang Kaisheng, Lin Mao. the spectral signature of Oligostilbenes, source of students and biological activity, Acta Pharmaceutica Sinica, 2002,37 (1): 69-74; Chen Li, Chen Jianbo. the pharmacological research summary of stilbene compound, Guangdong pharmacy, 2005,15 (3): 84-86]
Peony seed oil contains multiple unsaturated fatty acid, especially containing rare odd-numbered fatty acid, is a kind of edible oil and fat source with important Development volue.On March 22nd, 2011, Ministry of Health of the People's Republic of China approval from Paeonia ostii Paeonia suffruticosa (
paeonia ostiit.Hong et J.X.Zhang) and Paeonia papaveracea (
paeonia rockii) the peony seed oil made through techniques such as transmission from one meridian to another squeezing, decolouring, deodorizes of seed core can produce as new resource food and edible [Ministry of Health of the People's Republic of China announces, 2011 No. 9: Ministry of Public Health is about approval Acer Truncatum Buge seed oil and the peony seed oil bulletin as new resource food].This plays very large facilitation by for the production of peony seed oil and use.
Due to the high nutritive value of peony seed oil, the plantation amount of China's oil seed of Flos Moutan is increasing gradually, and the output of oil seed of Flos Moutan also increases year by year.At present, the preparation technology of peony seed oil is first by seed of Flos Moutan drying shelling.According to current output, oil seed of Flos Moutan output is 30,000 tons, the seed of Flos Moutan skin about shelling can produce about 3000-5000 ton, and the exploitation of these seed of Flos Moutan skins will be extremely important and research topics for arduous task.
In recent years, existing seminar is studied stilbene compound in medicinal plants seed of Flos Moutan: and document [He Chunnian. Paeonia Introduction of Pharmaphylogeny is studied, China Concord Medical Science University [D], 2010] separation process of 9 kinds of stilbene compounds is reported, the stilbene compound yield that it obtains is very low, maximum more than 100 milligrams of the monomer stilbene compound obtained from 4 kilograms of raw materials, is difficult to industrialization.In addition, its parting material used is seed of Paeonia suffruticosa, also there is the application problem of toxic organic solvents in research process.
This seminar early-stage Study shows, containing more oligomeric stilbene compound in Paeonia suffruticosa seedcake, further active testing shows that these stilbene compounds have good antibacterial activity.Therefore, from Paeonia suffruticosa seedcake, be separated stilbene compound, and study its antibacterial activity and have very important significance.
Summary of the invention
The object of this invention is to provide the compositions of the total stilbene compound of a kind of seed of Flos Moutan and extracting method thereof and application, the stilbene compound good anti-bacterial effect extracted from seed of Flos Moutan, applied range, natural component, safety is high.
The technical scheme that the present invention realizes above-mentioned purpose employing is: the extracting method of the compositions of the total stilbene compound of a kind of seed of Flos Moutan, comprises the following steps:
1) seed of Flos Moutan raw material is got, removing impurity, and with clean water 2-3 time, then thermostatic drying chamber is placed in, dry 3-9h under 60-80 DEG C of condition, add in hulling machine after oven dry and remove the peel, then the seed of Flos Moutan skin obtained is dried to water content and is no more than 10%, pulverize, be placed in extractor, adding seed of Flos Moutan tare weight amount 10-20 mass concentration is doubly the ethanol of 60-80%, reflux, extract, 8-10h under 40-60 DEG C of condition, extraction is filtered after terminating, and obtains soaking paste by filtering the filtrate evaporation and concentration obtained, for subsequent use;
2) leaching paste step 1) obtained its weight 3-5 organic solvent dissolution doubly, removing insoluble matter, then adding weight is leaching paste 3-6 adsorbent doubly, described adsorbent is mass ratio is the silica gel of 1:1-3 and diatomaceous mixture, the dry 4-12h in ventilation is placed on after stirring, obtain mixing, for subsequent use;
3) by step 2) mixing that obtains extracts with solvent petroleum ether-ethyl acetate, ethyl acetate, methanol successively, collects methanol extraction liquid, and concentratedly under 60 DEG C of conditions obtain total stilbene compound crude product being less than, for subsequent use;
4) total stilbene compound crude product step 3) obtained under 40-50 DEG C of condition with the acetone solution of its weight 2-4 times amount, dissolve completely and add the active carbon that weight is total stilbene compound crude product 4-7 times, filter after stirring 10-30min, mixing speed is 60-100r/min, collect filtrate and to be less than under 60 DEG C of conditions concentrated, be concentrated into solvent-free, namely obtain the compositions of total stilbene compound.
Described seed of Flos Moutan raw material is one or both of Paeonia ostii Paeonia suffruticosa seed or paeonia rockii seed.
In described step 1), seed of Flos Moutan skin reflux, extract, and filter process are at least in triplicate, merging filtrate.
Described step 2) in organic solvent be one or more of acetone, ethanol or ethyl acetate.
The volume ratio of the extractant petroleum ether-ethyl acetate in described step 3) is 1:1.
Each extraction solvent for use in described step 3) is 3-5 times of mixing weight.
The compositions of the described total stilbene compound of a kind of seed of Flos Moutan, comprises the stilbene compound of following percentage by weight:
Compound
1: Paeonia suffruticosa alcohol A, content is 10-15%;
Compound
2: Paeonia suffruticosa alcohol B, content is 22-28%;
Compound
3: Paeonia suffruticosa alcohol C, content is 10-20%;
Compound
4: cis Portugal root element, content is 0.5-1%;
Compound
5: trans Portugal root element, content is 2-5%;
Compound
6: Caulis Gneti H, content is 23-33%;
The structural formula of compound 1-6 is respectively:
。
The application of compositions in preparation antibacterials of the total stilbene compound of a kind of seed of Flos Moutan.
The application of compositions in food antibacterial of the total stilbene compound of a kind of seed of Flos Moutan.
Beneficial effect of the present invention
The compositions of one, the total stilbene compound of Paeonia suffruticosa of the present invention, prove through pharmacology test, there is the activity of good resisting gram-positive bacteria and negative bacterium, can as antibacterial active constituents of medicine, can be used alone, or be combined with suitable excipient, be used in food as food additive, or for the preparation of antibacterial medicine.
Two, extracting method of the present invention adopts ethanol extraction, the stilbene constituents that the ethanol of low concentration can make polarity relatively large is as much as possible to be extracted, extract at the temperature of 40-60 DEG C and both can ensure that effective ingredient proposed, the shortcoming of stilbene compound at high temperature poor stability can be avoided again, as long reflux, extract; Adopt and mix silica gel and the kieselguhr low price of sample, and solvent ethyl acetate and ethanol safety better, whole preparation process economical and convenient, be easy to industrialization.
Accompanying drawing explanation
Fig. 1 is the HPLC collection of illustrative plates of reference substance;
Fig. 2 is the HPLC collection of illustrative plates of the total stilbene compound of embodiment 1 gained;
Fig. 3 is the HPLC collection of illustrative plates of the total stilbene compound of embodiment 2 gained;
Fig. 4 is the HPLC collection of illustrative plates of the total stilbene compound of embodiment 3 gained;
The nuclear magnetic resonance, NMR IH-NMR collection of illustrative plates of Fig. 5 compound 1 (Paeonia suffruticosa alcohol A);
The nuclear magnetic resonance, NMR 13C-NMR collection of illustrative plates of Fig. 6 compound 1 (Paeonia suffruticosa alcohol A);
The nuclear magnetic resonance, NMR 1H-NMR collection of illustrative plates of Fig. 7 compound 2 (Paeonia suffruticosa alcohol B);
The nuclear magnetic resonance, NMR 13C-NMR collection of illustrative plates of Fig. 8 compound 2 (Paeonia suffruticosa alcohol B);
The nuclear magnetic resonance, NMR 1H-NMR collection of illustrative plates of Fig. 9 compound 3 (Paeonia suffruticosa alcohol C);
The nuclear magnetic resonance, NMR 13C-NMR collection of illustrative plates of Figure 10 compound 3 (Paeonia suffruticosa alcohol C);
The nuclear magnetic resonance, NMR 1H-NMR collection of illustrative plates of Figure 11 compound 4 (cis Portugal root element);
The nuclear magnetic resonance, NMR 13C-NMR collection of illustrative plates of Figure 12 compound 4 (cis Portugal root element);
The nuclear magnetic resonance, NMR 1H-NMR collection of illustrative plates of Figure 13 compound 5 (trans Portugal root element);
The nuclear magnetic resonance, NMR 13C-NMR collection of illustrative plates of Figure 14 compound 5 (trans Portugal root element);
The nuclear magnetic resonance, NMR 1H-NMR collection of illustrative plates of Figure 15 compound 6 (Caulis Gneti H);
The nuclear magnetic resonance, NMR 13C-NMR collection of illustrative plates of Figure 16 compound 6 (Caulis Gneti H).
Detailed description of the invention
An extracting method for the compositions of the total stilbene compound of seed of Flos Moutan, comprises the following steps:
1) seed of Flos Moutan raw material is got, removing impurity, and with clean water 2-3 time, then thermostatic drying chamber is placed in, dry 3-9h under 60-80 DEG C of condition, add in hulling machine after oven dry and remove the peel, then the seed of Flos Moutan skin obtained is dried to water content and is no more than 10%, pulverize, be placed in extractor, adding seed of Flos Moutan tare weight amount 10-20 mass concentration is doubly the ethanol of 60-80%, reflux, extract, 8-10h under 40-60 DEG C of condition, extraction is filtered after terminating, and obtains soaking paste by filtering the filtrate evaporation and concentration obtained, for subsequent use;
2) leaching paste step 1) obtained its weight 3-5 organic solvent dissolution doubly, removing insoluble matter, then adding weight is leaching paste 3-6 adsorbent doubly, described adsorbent is mass ratio is the silica gel of 1:1-3 and diatomaceous mixture, the dry 4-12h in ventilation is placed on after stirring, obtain mixing, for subsequent use;
3) by step 2) mixing that obtains extracts with solvent petroleum ether-ethyl acetate, ethyl acetate, methanol successively, collects methanol extraction liquid, and concentratedly under 60 DEG C of conditions obtain total stilbene compound crude product being less than, for subsequent use;
4) total stilbene compound crude product step 3) obtained under 40-50 DEG C of condition with the acetone solution of its weight 2-4 times amount, dissolve completely and add the active carbon that weight is total stilbene compound crude product 4-7 times, filter after stirring 10-30min, mixing speed is 60-100r/min, collect filtrate and to be less than under 60 DEG C of conditions concentrated, be concentrated into solvent-free, namely obtain the compositions of total stilbene compound.
Described seed of Flos Moutan raw material is one or both of Paeonia ostii Paeonia suffruticosa seed or paeonia rockii seed.
In described step 1), seed of Flos Moutan skin reflux, extract, and filter process are at least in triplicate, merging filtrate.
Described step 2) in organic solvent be one or more of acetone, ethanol or ethyl acetate.
The volume ratio of the extractant petroleum ether-ethyl acetate in described step 3) is 1:1.
Each extraction solvent for use in described step 3) is 3-5 times of mixing weight.
The compositions of the described total stilbene compound of a kind of seed of Flos Moutan, comprises the stilbene compound of following percentage by weight:
Compound
1: Paeonia suffruticosa alcohol A, content is 10-15%;
Compound
2: Paeonia suffruticosa alcohol B, content is 22-28%;
Compound
3: Paeonia suffruticosa alcohol C, content is 10-20%;
Compound
4: cis Portugal root element, content is 0.5-1%;
Compound
5: trans Portugal root element, content is 2-5%;
Compound
6: Caulis Gneti H, content is 23-33%;
The structural formula of compound 1-6 is respectively:
。
The application of compositions in preparation antibacterials of the total stilbene compound of a kind of seed of Flos Moutan.
The application of compositions in food antibacterial of the total stilbene compound of a kind of seed of Flos Moutan.
the detection chromatographic condition of stilbene compound
Agilent1100 high performance liquid chromatograph;
Chromatographic column: Zorbax SB C
18chromatographic column (4.6 × 250 mm, 5
);
Mobile phase: acetonitrile-potassium dihydrogen phosphate buffer salt gradient elution (45:55);
Flow: 1.0 mLmin
-1;
Determined wavelength: 230 nm;
Column temperature: 30 DEG C;
Sample size: 10
.
Through standard substance linearity curve, calculate the content of each stilbene compound successively.
Get standard sample and carry out high performance liquid chromatography detection, as shown in Figure 1, number corresponding compound in figure is gained collection of illustrative plates: 1:Suffruficosol A (Paeonia suffruticosa alcohol A); 2:Suffruficosol B (Paeonia suffruticosa alcohol B); 3:Suffruficosol C (Paeonia suffruticosa alcohol C); 4:Cis-e-Viniferin (cis Portugal root element); 5:Trans-e-Viniferin (trans Portugal root element); 6:Gnetin H (Caulis Gneti H).
The structural analysis of compound 1-6 can be schemed, as Fig. 5-16 see its nuclear magnetic resonance, NMR IH-NMR and 13C-NMR separately.
below in conjunction with specific embodiment, the present invention will be further described:
embodiment 1:
An extracting method for the compositions of the total stilbene compound of seed of Flos Moutan, comprises the following steps:
1) Paeonia ostii Paeonia suffruticosa seed raw material is got, removing impurity, and by clean water 2 times, then thermostatic drying chamber is placed in, dry 3h under 60 DEG C of conditions, add in hulling machine after oven dry and remove the peel, then the seed of Flos Moutan skin obtained is dried to water content and is no more than 10%, pulverize, get 100g and be placed in extractor, the mass concentration adding seed of Flos Moutan tare weight amount 10 times is the ethanol of 60%, reflux, extract, 8h under 40 DEG C of conditions, extracts after terminating and filters, repeat to extract and filter process three times, merging filtrate, obtains soaking paste by filtering the filtrate evaporation and concentration obtained, for subsequent use;
2) leaching paste step 1) the obtained acetone solvent of its weight 3 times dissolves, removing insoluble matter, then the adsorbent that weight is leaching paste 3 times is added, described adsorbent is mass ratio is the silica gel of 1:1 and diatomaceous mixture, the dry 4h in ventilation is placed on after stirring, obtain mixing, for subsequent use;
3) by step 2) mixing that obtains extracts with solvent petroleum ether-ethyl acetate, ethyl acetate, methanol that quality multiple is 3 times successively, the volume ratio of petroleum ether-ethyl acetate is 1:1, collect methanol extraction liquid, and concentratedly obtain total stilbene compound crude product being less than under 60 DEG C of conditions, for subsequent use;
4) total stilbene compound crude product step 3) obtained under 40 DEG C of conditions with the acetone solution of its weight 2 times amount, dissolve completely and add the active carbon that weight is total stilbene compound crude product 4 times, filter after stirring 10min, mixing speed is 60r/min, collect filtrate and to be less than under 60 DEG C of conditions concentrated, be concentrated into solvent-free, namely obtain the compositions of total stilbene compound, altogether 0.9g.
After testing, the compositions of total stilbene compound that the present embodiment obtains comprises the stilbene compound of following percentage by weight:
Compound 1: Paeonia suffruticosa alcohol A (Suffruficosol A), content is 10.5%;
Compound 2: Paeonia suffruticosa alcohol B (Suffruficosol B), content is 27.3%;
Compound 3: Paeonia suffruticosa alcohol C (Suffruficosol C), content is 19.6%;
Compound 4: cis Portugal root element (Cis-e-Viniferin), content is 0.8%;
Compound 5: trans Portugal's root element (Trans-e-Viniferin), content is 2.6%;
Compound 6: Caulis Gneti H (Gnetin H), content is 23.8%;
The content 83.6% of total stilbene compound;
The compositions of total stilbene compound that Example 1 obtains carries out high performance liquid chromatography detection, and as shown in Figure 2, in figure, the numeral of labelling is corresponding with compound for gained collection of illustrative plates.
embodiment 2:
An extracting method for the compositions of the total stilbene compound of seed of Flos Moutan, comprises the following steps:
1) Paeonia papaveracea seed raw material is got, removing impurity, and by clean water 3 times, then thermostatic drying chamber is placed in, dry 9h under 80 DEG C of conditions, add in hulling machine after oven dry and remove the peel, then the seed of Flos Moutan skin obtained is dried to water content and is no more than 10%, pulverize, get 300g and be placed in extractor, the mass concentration adding seed of Flos Moutan tare weight amount 20 times is the ethanol of 80%, reflux, extract, 10h under 60 DEG C of conditions, extracts after terminating and filters, repeat to extract and filter process five times, merging filtrate, obtains soaking paste by filtering the filtrate evaporation and concentration obtained, for subsequent use;
2) leaching paste step 1) the obtained alcohol solvent of its weight 5 times dissolves, removing insoluble matter, then the adsorbent that weight is leaching paste 6 times is added, described adsorbent is mass ratio is the silica gel of 1:3 and diatomaceous mixture, the dry 12h in ventilation is placed on after stirring, obtain mixing, for subsequent use;
3) by step 2) mixing that obtains extracts with solvent petroleum ether-ethyl acetate, ethyl acetate, methanol that quality multiple is 5 times successively, the volume ratio of petroleum ether-ethyl acetate is 1:1, collect methanol extraction liquid, and concentratedly obtain total stilbene compound crude product being less than under 60 DEG C of conditions, for subsequent use;
4) total stilbene compound crude product step 3) obtained under 50 DEG C of conditions with the acetone solution of its weight 4 times amount, dissolve completely and add the active carbon that weight is total stilbene compound crude product 7 times, filter after stirring 30min, mixing speed is 100r/min, collect filtrate and to be less than under 60 DEG C of conditions concentrated, be concentrated into solvent-free, namely obtain the compositions of total stilbene compound, altogether 3.1g.
After testing, the compositions of the total stilbene compound of seed of Flos Moutan that the present embodiment obtains comprises the stilbene compound of following percentage by weight:
Compound 1: Paeonia suffruticosa alcohol A (Suffruficosol A), content is 14.6%;
Compound 2: Paeonia suffruticosa alcohol B (Suffruficosol B), content is 22.3%;
Compound 3: Paeonia suffruticosa alcohol C (Suffruficosol C), content is 14.1%;
Compound 4: cis Portugal root element (Cis-e-Viniferin), content is 0.9%;
Compound 5: trans Portugal's root element (Trans-e-Viniferin), content is 3.1%;
Compound 6: Caulis Gneti H (Gnetin H), content is 32.4%.
The content 87.4% of total stilbene compound;
The compositions of total stilbene compound that Example 2 obtains carries out high performance liquid chromatography detection, and as shown in Figure 3, in figure, the numeral of labelling is corresponding with compound for gained collection of illustrative plates.
embodiment 3:
An extracting method for the compositions of the total stilbene compound of seed of Flos Moutan, comprises the following steps:
1) getting mass ratio is the Paeonia ostii Paeonia suffruticosa seed of 1:1 and the mixed material of paeonia rockii seed, removing impurity, and by clean water 3 times, then thermostatic drying chamber is placed in, dry 6h under 70 DEG C of conditions, add in hulling machine after oven dry and remove the peel, then the seed of Flos Moutan skin obtained is dried to water content and is no more than 10%, pulverize, get 100g and be placed in extractor, the mass concentration adding seed of Flos Moutan tare weight amount 15 times is the ethanol of 70%, reflux, extract, 9h under 50 DEG C of conditions, extraction is filtered after terminating, repeat to extract and filter process six times, merging filtrate, obtain soaking paste by filtering the filtrate evaporation and concentration obtained, for subsequent use,
2) leaching paste step 1) the obtained ethyl acetate solvent of its weight 4 times dissolves, removing insoluble matter, then the adsorbent that weight is leaching paste 5 times is added, described adsorbent is mass ratio is the silica gel of 1:2 and diatomaceous mixture, the dry 8h in ventilation is placed on after stirring, obtain mixing, for subsequent use;
3) by step 2) mixing that obtains extracts with solvent petroleum ether-ethyl acetate, ethyl acetate, methanol that quality multiple is 4 successively, the volume ratio of petroleum ether-ethyl acetate is 1:1, collect methanol extraction liquid, and concentratedly obtain total stilbene compound crude product being less than under 60 DEG C of conditions, for subsequent use;
4) total stilbene compound crude product step 3) obtained under 40-50 DEG C of condition with the acetone solution of its weight 3 times amount, dissolve completely and add the active carbon that weight is total stilbene compound crude product 5 times, filter after stirring 20min, mixing speed is 80r/min, collect filtrate and to be less than under 60 DEG C of conditions concentrated, be concentrated into solvent-free, namely obtain the compositions of total stilbene compound, altogether 0.85g.
After testing, the compositions of the total stilbene compound of seed of Flos Moutan that the present embodiment obtains comprises the stilbene compound of following percentage by weight:
Compound 1: Paeonia suffruticosa alcohol A (Suffruficosol A), content is 14.1%;
Compound 2: Paeonia suffruticosa alcohol B (Suffruficosol B), content is 26.4%;
Compound 3: Paeonia suffruticosa alcohol C (Suffruficosol C), content is 10.5%;
Compound 4: cis Portugal root element (Cis-e-Viniferin), content is 0.6%;
Compound 5: trans Portugal's root element (Trans-e-Viniferin), content is 4.6%;
Compound 6: Caulis Gneti H (Gnetin H), content is 27.9%.
The content 84.1% of total stilbene compound;
Surplus is the stilbene compound of not yet clear and definite structure and inevitable impurity;
The compositions of total stilbene compound that Example 2 obtains carries out high performance liquid chromatography detection, and as shown in Figure 3, in figure, the numeral of labelling is corresponding with compound for gained collection of illustrative plates.
pharmacological testing
1. test material
Seed of Flos Moutan skin stilbene compound compositions (three groups of compositionss that above-mentioned three groups of embodiments obtain are respectively compositions 1, compositions 2, compositions 3);
Bacterial strain: staphylococcus aureus
s. aureas, streptococcus uberis
s. uberis, bacillus subtilis
bacillus subtilis, escherichia coli
e. agalactiae, Salmonella
samonellalthered is provided by The First Affiliated Hospital of Henan University of Science and Technology.
2. test method
The mensuration of minimum inhibitory concentration (MIC):
Adopt document [Li Jianqiang, Li Liujin. veterinary microbiology test practice instructs [M]. Xi'an: Shaanxi science tech publishing house, 1999:29-30] middle test tube turbidimetry, by the sterilizing test tubes numbering of 10 band tampons, No. 1 pipe adds cultured solution of broth 1.5 mL, and all the other each pipes add 1 mL.Get 10 mg/mL sample solutions (solvent is the mixed liquor of V (methanol): V (water)=1: 1) 0.5 mL to add in No. 1 pipe and mix, from No. 1 pipe, get 1 mL again add No. 2 pipes, serial dilution to like this No. 8 pipe, No. 8 pipes discard 1 mL containing medicinal liquid meat soup.Draw 50
bacteria containing amount is equivalent to the bacterial suspension (bacterial concentration 1.5 hundred million/mL) of Maxwell opacity tube the 1st pipe 1/2 to 1 ~ No. 9 pipe, and No. 10 pipes add 50
meat soup.Take out after putting 37 DEG C of cultivation 12 ~ 15 h and observe turbidity.No. 9 pipes are muddy positive control pipe, and No. 10 pipes are muddy negative control pipe.Manage than No. 9 limpid and respectively manage thereafter the test tube consistent with No. 9 pipe turbidity, its medicament contg is minimum inhibitory concentration.
result of the test
Research finds that seed of Flos Moutan skin stilbene compound total extract is to gram positive bacteria (staphylococcus aureus, streptococcus uberis, bacillus subtilis) and gram negative bacteria (escherichia coli, Salmonella) all there is good inhibitory action, and drug effect is relatively better.Experimental result is as follows:
Table seed of Flos Moutan stilbene compound compositions minimum inhibitory concentration (MIC) (
/ mL)
。
Claims (8)
1. an extracting method for the compositions of the total stilbene compound of seed of Flos Moutan, is characterized in that: comprise the following steps:
1) seed of Flos Moutan raw material is got, removing impurity, and with clean water 2-3 time, then thermostatic drying chamber is placed in, dry 3-9h under 60-80 DEG C of condition, add in hulling machine after oven dry and remove the peel, then the seed of Flos Moutan skin obtained is dried to water content and is no more than 10%, pulverize, be placed in extractor, adding seed of Flos Moutan tare weight amount 10-20 mass concentration is doubly the ethanol of 60-80%, reflux, extract, 8-10h under 40-60 DEG C of condition, extraction is filtered after terminating, and obtains soaking paste by filtering the filtrate evaporation and concentration obtained, for subsequent use;
2) leaching paste step 1) obtained its weight 3-5 organic solvent dissolution doubly, removing insoluble matter, organic solvent is one or more of acetone, ethanol or ethyl acetate; Then adding weight is leaching paste 3-6 adsorbent doubly, and described adsorbent is mass ratio is the silica gel of 1:1-3 and diatomaceous mixture, is placed on the dry 4-12h in ventilation, obtains mixing after stirring, for subsequent use;
3) by step 2) mixing that obtains extracts with solvent petroleum ether-ethyl acetate, ethyl acetate, methanol successively, collects methanol extraction liquid, and concentratedly under 60 DEG C of conditions obtain total stilbene compound crude product being less than, for subsequent use;
4) total stilbene compound crude product step 3) obtained under 40-50 DEG C of condition with the acetone solution of its weight 2-4 times amount, dissolve completely and add the active carbon that weight is total stilbene compound crude product 4-7 times, filter after stirring 10-30min, mixing speed is 60-100r/min, collect filtrate and to be less than under 60 DEG C of conditions concentrated, be concentrated into solvent-free, namely obtain the compositions of total stilbene compound.
2. the extracting method of the compositions of the total stilbene compound of a kind of seed of Flos Moutan as claimed in claim 1, is characterized in that: described seed of Flos Moutan raw material is one or both of Paeonia ostii Paeonia suffruticosa seed or paeonia rockii seed.
3. the extracting method of the compositions of the total stilbene compound of a kind of seed of Flos Moutan as claimed in claim 1, is characterized in that: in described step 1), seed of Flos Moutan skin reflux, extract, and filter process are at least in triplicate, merging filtrate.
4. the extracting method of the compositions of the total stilbene compound of a kind of seed of Flos Moutan as claimed in claim 1, is characterized in that: the volume ratio of the extractant petroleum ether-ethyl acetate in described step 3) is 1:1.
5. the extracting method of the compositions of the total stilbene compound of a kind of seed of Flos Moutan as claimed in claim 1, is characterized in that: each extraction solvent for use in described step 3) is 3-5 times of mixing weight.
6. the compositions of the total stilbene compound of a kind of seed of Flos Moutan as claimed in claim 1, is characterized in that: the stilbene compound comprising following percentage by weight:
Compound
1: Paeonia suffruticosa alcohol A, content is 10-15%;
Compound
2: Paeonia suffruticosa alcohol B, content is 22-28%;
Compound
3: Paeonia suffruticosa alcohol C, content is 10-20%;
Compound
4: cis Portugal root element, content is 0.5-1%;
Compound
5: trans Portugal root element, content is 2-5%;
Compound
6: Caulis Gneti H, content is 23-33%;
The structural formula of compound 1-6 is respectively:
。
7. the application of compositions in preparation antibacterials of the total stilbene compound of a kind of seed of Flos Moutan as claimed in claim 6.
8. the application of compositions in food antibacterial of the total stilbene compound of a kind of seed of Flos Moutan as claimed in claim 6.
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| CN103772334B (en) * | 2014-01-10 | 2015-07-08 | 河南科技大学 | Method for extracting stilbene compound from penoy seed husk |
| CN107412221A (en) * | 2017-08-03 | 2017-12-01 | 西北大学 | A kind of tree peony seed extract and its application as antineoplastic |
| CN107361203B (en) * | 2017-08-08 | 2020-07-14 | 河南科技大学 | Preparation method and application of natural anti-feed granules |
| CN108078885B (en) * | 2018-02-05 | 2020-09-18 | 海南师范大学 | Dendrobium extract and application thereof in preparation of whitening sunscreen agent |
| CN112538066B (en) * | 2020-12-14 | 2023-11-10 | 中国科学院植物研究所 | Paeonia lactiflora seed coat extract for resisting skin cell aging |
| CN113509421A (en) * | 2021-08-19 | 2021-10-19 | 河南科技大学 | Peony seed shell cosmetic sunscreen raw material and application thereof |
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