CN103494797A - 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 total stilbene compound compositions of a kind of seed of Flos Moutan and extracting method and application.
Background technology
Stilbene compound (stilbeniods) refers to monomer with 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 contains rare odd-numbered fatty acid, is a kind of edible oil and fat source that important exploitation is worth that has.On March 22nd, 2011, Ministry of Health of the People's Republic of China approval from the 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 as new resource food, can produce and edible [Ministry of Health of the People's Republic of China's bulletin, 2011 No. 9: Ministry of Public Health is the bulletin as new resource food about approval Acer Truncatum Buge seed oil and peony seed oil].This will play very large facilitation for production and the use of peony seed oil.
Due to the high nutritive value of peony seed oil, China's oil is increasing gradually by the plantation amount of seed of Flos Moutan, and oil also increases year by year by the output of seed of Flos Moutan.At present, the preparation technology of peony seed oil is first by the dry shelling of seed of Flos Moutan.According to current output, oil is 30,000 tons by seed of Flos Moutan output, can produce the seed of Flos Moutan skin of about 3000-5000 ton left and right through shelling, and the exploitation of these seed of Flos Moutan skins will be the research topics of an extremely important and arduous task.
In recent years, existing seminar is studied stilbene compound in the medicinal plants seed of Flos Moutan: and document [He Chunnian. the research of Paeonia Introduction of Pharmaphylogeny, China Concord Medical Science University [D], 2010] reported the separation process of 9 kinds of stilbene compounds, its resulting stilbene compound yield is very low, from 4 kilograms of raw materials, resulting monomer stilbene compound is maximum more than 100 milligrams, is difficult to industrialization.In addition, its parting material used is seed of Paeonia suffruticosa, also exists the application problem of poisonous organic solvent in research process.
This seminar early-stage Study shows, contains more oligomeric stilbene compound in Paeonia suffruticosa seedcake, and further active testing shows that these stilbene compounds have antibacterial activity preferably.Therefore, from Paeonia suffruticosa seedcake, separate stilbene compound, and study its antibacterial activity and have very important significance.
Summary of the invention
The purpose of this invention is to provide the total stilbene compound compositions of a kind of seed of Flos Moutan and extracting method and application, the stilbene compound good anti-bacterial effect extracted from seed of Flos Moutan, applied range, natural component, safe.
The present invention realizes that the technical scheme that above-mentioned purpose adopts is: the extracting method of the total stilbene compound compositions of a kind of seed of Flos Moutan comprises the following steps:
1) get the seed of Flos Moutan raw material, remove impurity, and clean 2-3 time with clear water, then be placed in thermostatic drying chamber, dry 3-9h under 60-80 ℃ of condition, add in hulling machine and remove the peel after oven dry, then the seed of Flos Moutan skin obtained is dried to water content and is no more than 10%, pulverize, be placed in extractor, add the ethanol that seed of Flos Moutan tare weight amount 10-20 mass concentration doubly is 60-80%, reflux, extract, 8-10h under 40-60 ℃ of condition, after extracting end, filtered, the filtrate evaporation and concentration that filtration is obtained obtains soaking paste, standby;
2) step 1) is obtained soak paste with its weight 3-5 organic solvent dissolution doubly, remove insoluble matter, then adding weight is to soak paste 3-6 adsorbent doubly, described adsorbent is silica gel and the diatomaceous mixture that mass ratio is 1:1-3, be placed on the dry 4-12h in ventilation after stirring, obtain mixing, standby;
3) by step 2) mixing that obtains extracted with solvent petroleum ether-ethyl acetate, ethyl acetate, methanol successively, collects methanol extraction liquid, and concentratedly under 60 ℃ of conditions obtain total stilbene compound crude product being less than, standby;
4) acetone solution that total stilbene compound crude product step 3) obtained is doubly measured with its weight 2-4 under 40-50 ℃ of condition, adding weight after dissolve complete is total stilbene compound crude product 4-7 active carbon doubly, after stirring 10-30min, filter, mixing speed is 60-100r/min, collect filtrate and be less than under 60 ℃ of conditions concentrated, be concentrated into solvent-freely, obtain total stilbene compound compositions.
Described seed of Flos Moutan raw material is one or both of Paeonia ostii seed of Flos Moutan or paeonia rockii seed.
Seed of Flos Moutan skin reflux, extract, and filter process triplicate at least in described step 1), merging filtrate.
Described step 2) organic solvent in is 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.
The 3-5 that each extraction solvent for use in described step 3) is mixing weight doubly.
The total stilbene compound compositions of described 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 the total stilbene compound compositions of a kind of seed of Flos Moutan in the preparation antibacterials.
The application of the total stilbene compound compositions of a kind of seed of Flos Moutan in the food antibacterial.
Beneficial effect of the present invention
One, the total stilbene compound compositions of Paeonia suffruticosa of the present invention, through pharmacology test, prove, there is the activity of resisting gram-positive bacteria and negative bacterium preferably, can be used as antibiotic active constituents of medicine, can use separately, or be combined with suitable excipient, be used for food as food additive, or for the preparation of antibiotic medicine.
Two, extracting method of the present invention adopts ethanol extraction, the ethanol of low concentration can make polar phase to larger as much as possible being extracted of stilbene constituents, extract at the temperature of 40-60 ℃ and both can guarantee that effective ingredient proposed, can avoid again the at high temperature shortcoming of poor stability of stilbene compound, as long reflux, extract; Silica gel and the kieselguhr low price of sample mixed in employing, and solvent ethyl acetate and the ethanol safety better, whole preparation process economical and convenient, be easy to industrialization.
The accompanying drawing explanation
The HPLC collection of illustrative plates that Fig. 1 is reference substance;
The HPLC collection of illustrative plates that Fig. 2 is the total stilbene compound of embodiment 1 gained;
The HPLC collection of illustrative plates that Fig. 3 is the total stilbene compound of embodiment 2 gained;
The HPLC collection of illustrative plates that Fig. 4 is 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).
The specific embodiment
The extracting method of the total stilbene compound compositions of a kind of seed of Flos Moutan comprises the following steps:
1) get the seed of Flos Moutan raw material, remove impurity, and clean 2-3 time with clear water, then be placed in thermostatic drying chamber, dry 3-9h under 60-80 ℃ of condition, add in hulling machine and remove the peel after oven dry, then the seed of Flos Moutan skin obtained is dried to water content and is no more than 10%, pulverize, be placed in extractor, add the ethanol that seed of Flos Moutan tare weight amount 10-20 mass concentration doubly is 60-80%, reflux, extract, 8-10h under 40-60 ℃ of condition, after extracting end, filtered, the filtrate evaporation and concentration that filtration is obtained obtains soaking paste, standby;
2) step 1) is obtained soak paste with its weight 3-5 organic solvent dissolution doubly, remove insoluble matter, then adding weight is to soak paste 3-6 adsorbent doubly, described adsorbent is silica gel and the diatomaceous mixture that mass ratio is 1:1-3, be placed on the dry 4-12h in ventilation after stirring, obtain mixing, standby;
3) by step 2) mixing that obtains extracted with solvent petroleum ether-ethyl acetate, ethyl acetate, methanol successively, collects methanol extraction liquid, and concentratedly under 60 ℃ of conditions obtain total stilbene compound crude product being less than, standby;
4) acetone solution that total stilbene compound crude product step 3) obtained is doubly measured with its weight 2-4 under 40-50 ℃ of condition, adding weight after dissolve complete is total stilbene compound crude product 4-7 active carbon doubly, after stirring 10-30min, filter, mixing speed is 60-100r/min, collect filtrate and be less than under 60 ℃ of conditions concentrated, be concentrated into solvent-freely, obtain total stilbene compound compositions.
Described seed of Flos Moutan raw material is one or both of Paeonia ostii seed of Flos Moutan or paeonia rockii seed.
Seed of Flos Moutan skin reflux, extract, and filter process triplicate at least in described step 1), merging filtrate.
Described step 2) organic solvent in is 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.
The 3-5 that each extraction solvent for use in described step 3) is mixing weight doubly.
The total stilbene compound compositions of described 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 the total stilbene compound compositions of a kind of seed of Flos Moutan in the preparation antibacterials.
The application of the total stilbene compound compositions of a kind of seed of Flos Moutan in the food antibacterial.
the detection chromatographic condition of stilbene compound
The 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;
Detect wavelength: 230 nm;
Column temperature: 30 ℃;
Sample size: 10
.
Through the standard substance linearity curve, calculate successively the content of each stilbene compound.
The accurate sample of label taking carries out the high performance liquid chromatography detection, and as shown in Figure 1, in figure, compound corresponding to numbering is the 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 referring to it nuclear magnetic resonance, NMR IH-NMR and 13C-NMR figure separately, as Fig. 5-16.
below in conjunction with specific embodiment, the present invention will be further described:
embodiment 1:
The extracting method of the total stilbene compound compositions of a kind of seed of Flos Moutan comprises the following steps:
1) get Paeonia ostii seed of Flos Moutan raw material, remove impurity, and clean 2 times with clear water, then be placed in thermostatic drying chamber, dry 3h under 60 ℃ of conditions, add in hulling machine and remove the peel after oven dry, 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 ethanol that the mass concentration that adds 10 times of seed of Flos Moutan tare weight amounts is 60%, reflux, extract, 8h under 40 ℃ of conditions, extraction is filtered after finishing, repeat extraction and filter process three times, merging filtrate, the filtrate evaporation and concentration that filtration is obtained obtains soaking paste, standby;
2) paste that soaks step 1) obtained dissolves with the acetone solvent of 3 times of its weight, remove insoluble matter, then adding weight is to soak the adsorbent of 3 times of pastes, described adsorbent is silica gel and the diatomaceous mixture that mass ratio is 1:1, be placed on the dry 4h in ventilation after stirring, obtain mixing, standby;
3) by step 2) mixing that obtains is 3 times with the quality multiple solvent petroleum ether-ethyl acetate, ethyl acetate, methanol are extracted 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 ℃ of conditions, standby;
4) total stilbene compound crude product step 3) obtained under 40 ℃ of conditions with the acetone solution of 2 times of amounts of its weight, add the active carbon that weight is 4 times of total stilbene compound crude products after dissolve complete, after stirring 10min, filter, mixing speed is 60r/min, collect filtrate and be less than under 60 ℃ of conditions concentrated, be concentrated into solvent-freely, obtain total stilbene compound compositions, altogether 0.9g.
After testing, total stilbene compound compositions 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 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;
Get total stilbene compound compositions that embodiment 1 obtains and carry out the high performance liquid chromatography detection, as shown in Figure 2, in figure, the numeral of labelling is corresponding with compound for the gained collection of illustrative plates.
embodiment 2:
The extracting method of the total stilbene compound compositions of a kind of seed of Flos Moutan comprises the following steps:
1) get the Paeonia papaveracea seed raw material, remove impurity, and clean 3 times with clear water, then be placed in thermostatic drying chamber, dry 9h under 80 ℃ of conditions, add in hulling machine and remove the peel after oven dry, 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 ethanol that the mass concentration that adds 20 times of seed of Flos Moutan tare weight amounts is 80%, reflux, extract, 10h under 60 ℃ of conditions, extraction is filtered after finishing, repeat extraction and filter process five times, merging filtrate, the filtrate evaporation and concentration that filtration is obtained obtains soaking paste, standby;
2) paste that soaks step 1) obtained dissolves with the alcohol solvent of 5 times of its weight, remove insoluble matter, then adding weight is to soak the adsorbent of 6 times of pastes, described adsorbent is silica gel and the diatomaceous mixture that mass ratio is 1:3, be placed on the dry 12h in ventilation after stirring, obtain mixing, standby;
3) by step 2) mixing that obtains is 5 times with the quality multiple solvent petroleum ether-ethyl acetate, ethyl acetate, methanol are extracted 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 ℃ of conditions, standby;
4) total stilbene compound crude product step 3) obtained under 50 ℃ of conditions with the acetone solution of 4 times of amounts of its weight, add the active carbon that weight is 7 times of total stilbene compound crude products after dissolve complete, after stirring 30min, filter, mixing speed is 100r/min, collect filtrate and be less than under 60 ℃ of conditions concentrated, be concentrated into solvent-freely, obtain total stilbene compound compositions, altogether 3.1g.
After testing, the total stilbene compound compositions of the 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 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;
Get total stilbene compound compositions that embodiment 2 obtains and carry out the high performance liquid chromatography detection, as shown in Figure 3, in figure, the numeral of labelling is corresponding with compound for the gained collection of illustrative plates.
embodiment 3:
The extracting method of the total stilbene compound compositions of a kind of seed of Flos Moutan comprises the following steps:
1) get Paeonia ostii seed of Flos Moutan that mass ratio is 1:1 and the mixed material of paeonia rockii seed, remove impurity, and clean 3 times with clear water, then be placed in thermostatic drying chamber, dry 6h under 70 ℃ of conditions, add in hulling machine and remove the peel after oven dry, 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 ethanol that the mass concentration that adds 15 times of seed of Flos Moutan tare weight amounts is 70%, reflux, extract, 9h under 50 ℃ of conditions, after extracting end, filtered, repeat extraction and filter process six times, merging filtrate, the filtrate evaporation and concentration that filtration is obtained obtains soaking paste, standby,
2) paste that soaks step 1) obtained dissolves with the ethyl acetate solvent of 4 times of its weight, remove insoluble matter, then adding weight is to soak the adsorbent of 5 times of pastes, described adsorbent is silica gel and the diatomaceous mixture that mass ratio is 1:2, be placed on the dry 8h in ventilation after stirring, obtain mixing, standby;
3) by step 2) mixing that obtains is 4 with the quality multiple solvent petroleum ether-ethyl acetate, ethyl acetate, methanol are extracted 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 ℃ of conditions, standby;
4) total stilbene compound crude product step 3) obtained under 40-50 ℃ of condition with the acetone solution of 3 times of amounts of its weight, add the active carbon that weight is 5 times of total stilbene compound crude products after dissolve complete, after stirring 20min, filter, mixing speed is 80r/min, collect filtrate and be less than under 60 ℃ of conditions concentrated, be concentrated into solvent-freely, obtain total stilbene compound compositions, altogether 0.85g.
After testing, the total stilbene compound compositions of the 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 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;
The stilbene compound that surplus is not yet clear and definite structure and inevitable impurity;
Get total stilbene compound compositions that embodiment 2 obtains and carry out the high performance liquid chromatography detection, as shown in Figure 3, in figure, the numeral of labelling is corresponding with compound for the 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, be 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
samonellalby The First Affiliated Hospital of Henan University of Science and Technology, provided.
2. test method
The mensuration of minimum inhibitory concentration (MIC):
Adopt document [Li Jianqiang, Li Liujin. the veterinary microbiology test practice instructs [M]. Xi'an: Shaanxi science tech publishing house, 1999:29-30] middle test tube turbidimetry, by 10 numberings of the sterilizing test tubes with tampon, No. 1 pipe adds cultured solution of broth 1.5 mL, and all the other each pipes add 1 mL.Getting 10 mg/mL sample solutions (mixed liquor that solvent is V (methanol): V (water)=1: 1), 0.5 mL adds in No. 1 pipe and mixes, get again 1 mL and add pipe No. 2 from No. 1 pipe, so serial dilution to No. 8 is managed, and 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 pipe adds 50
meat soup.Take out the observation turbidity after putting 37 ℃ of cultivation 12～15 h.No. 9 pipes are muddy positive control pipe, and No. 10 pipes are muddy negative control pipe.Ratio is managed for No. 9 limpid and is respectively managed thereafter and manage with No. 9 the test tube that turbidity is consistent, and its medicament contg is minimum inhibitory concentration.
3. 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 inhibitory action preferably, 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 (9)
1. the extracting method of the total stilbene compound compositions of seed of Flos Moutan is characterized in that: comprise the following steps:
1) get the seed of Flos Moutan raw material, remove impurity, and clean 2-3 time with clear water, then be placed in thermostatic drying chamber, dry 3-9h under 60-80 ℃ of condition, add in hulling machine and remove the peel after oven dry, then the seed of Flos Moutan skin obtained is dried to water content and is no more than 10%, pulverize, be placed in extractor, add the ethanol that seed of Flos Moutan tare weight amount 10-20 mass concentration doubly is 60-80%, reflux, extract, 8-10h under 40-60 ℃ of condition, after extracting end, filtered, the filtrate evaporation and concentration that filtration is obtained obtains soaking paste, standby;
2) step 1) is obtained soak paste with its weight 3-5 organic solvent dissolution doubly, remove insoluble matter, then adding weight is to soak paste 3-6 adsorbent doubly, described adsorbent is silica gel and the diatomaceous mixture that mass ratio is 1:1-3, be placed on the dry 4-12h in ventilation after stirring, obtain mixing, standby;
3) by step 2) mixing that obtains extracted with solvent petroleum ether-ethyl acetate, ethyl acetate, methanol successively, collects methanol extraction liquid, and concentratedly under 60 ℃ of conditions obtain total stilbene compound crude product being less than, standby;
4) acetone solution that total stilbene compound crude product step 3) obtained is doubly measured with its weight 2-4 under 40-50 ℃ of condition, adding weight after dissolve complete is total stilbene compound crude product 4-7 active carbon doubly, after stirring 10-30min, filter, mixing speed is 60-100r/min, collect filtrate and be less than under 60 ℃ of conditions concentrated, be concentrated into solvent-freely, obtain total stilbene compound compositions.
2. the extracting method of the total stilbene compound compositions of a kind of seed of Flos Moutan as claimed in claim 1, it is characterized in that: described seed of Flos Moutan raw material is one or both of Paeonia ostii seed of Flos Moutan or paeonia rockii seed.
3. the extracting method of the total stilbene compound compositions of a kind of seed of Flos Moutan as claimed in claim 1 is characterized in that: seed of Flos Moutan skin reflux, extract, and filter process triplicate at least in described step 1), merging filtrate.
4. the extracting method of the total stilbene compound compositions of a kind of seed of Flos Moutan as claimed in claim 1, it is characterized in that: the organic solvent described step 2) is one or more of acetone, ethanol or ethyl acetate.
5. the extracting method of the total stilbene compound compositions of a kind of seed of Flos Moutan as claimed in claim 1, it is characterized in that: the volume ratio of the extractant petroleum ether-ethyl acetate in described step 3) is 1:1.
6. the extracting method of the total stilbene compound compositions of a kind of seed of Flos Moutan as claimed in claim 1, is characterized in that: the 3-5 that the each extraction solvent for use in described step 3) is mixing weight times.
7. the total stilbene compound compositions of a kind of seed of Flos Moutan as claimed in claim 1 is characterized in that: the stilbene compound that comprises 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:
8. the application of the total stilbene compound compositions of a kind of seed of Flos Moutan as claimed in claim 7 in the preparation antibacterials.
9. the application of the total stilbene compound compositions of a kind of seed of Flos Moutan as claimed in claim 7 in the food antibacterial.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103766913A (en) * | 2014-01-10 | 2014-05-07 | 河南科技大学 | Method for extracting total stilbene compound from penoy seeds |
| CN103772334A (en) * | 2014-01-10 | 2014-05-07 | 河南科技大学 | Method for extracting stilbene compound from penoy seed husk |
| CN107361203A (en) * | 2017-08-08 | 2017-11-21 | 河南科技大学 | It is a kind of naturally to replace anti-feed granules agent and its preparation method and application |
| CN107412221A (en) * | 2017-08-03 | 2017-12-01 | 西北大学 | A kind of tree peony seed extract and its application as antineoplastic |
| CN108078885A (en) * | 2018-02-05 | 2018-05-29 | 海南师范大学 | A kind of dendrobium extract and its application in whitening sunscreen agent is prepared |
| CN112538066A (en) * | 2020-12-14 | 2021-03-23 | 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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| CN103130850A (en) * | 2013-02-27 | 2013-06-05 | 河南科技大学 | Method for preparing paeoniflorin from oil peony seed meal |
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| CN103130850A (en) * | 2013-02-27 | 2013-06-05 | 河南科技大学 | Method for preparing paeoniflorin from oil peony seed meal |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103766913A (en) * | 2014-01-10 | 2014-05-07 | 河南科技大学 | Method for extracting total stilbene compound from penoy seeds |
| CN103772334A (en) * | 2014-01-10 | 2014-05-07 | 河南科技大学 | Method for extracting stilbene compound from penoy seed husk |
| CN103772334B (en) * | 2014-01-10 | 2015-07-08 | 河南科技大学 | Method for extracting stilbene compound from penoy seed husk |
| CN103766913B (en) * | 2014-01-10 | 2016-08-17 | 河南科技大学 | A kind of method extracting total stilbene compound from seed of Flos Moutan |
| CN107412221A (en) * | 2017-08-03 | 2017-12-01 | 西北大学 | A kind of tree peony seed extract and its application as antineoplastic |
| CN107361203A (en) * | 2017-08-08 | 2017-11-21 | 河南科技大学 | It is a kind of naturally to replace anti-feed granules agent and its preparation method and application |
| CN108078885A (en) * | 2018-02-05 | 2018-05-29 | 海南师范大学 | A kind of dendrobium extract and its application in whitening sunscreen agent is prepared |
| CN112538066A (en) * | 2020-12-14 | 2021-03-23 | 中国科学院植物研究所 | Paeonia lactiflora seed coat extract for resisting skin cell aging |
| 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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