CN108690724A - A kind of application of ford dutchmanspipe root or herb volatile oil in preparing anti-curing cancers and/antibacterial product - Google Patents
A kind of application of ford dutchmanspipe root or herb volatile oil in preparing anti-curing cancers and/antibacterial product Download PDFInfo
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- CN108690724A CN108690724A CN201810449997.7A CN201810449997A CN108690724A CN 108690724 A CN108690724 A CN 108690724A CN 201810449997 A CN201810449997 A CN 201810449997A CN 108690724 A CN108690724 A CN 108690724A
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- Prior art keywords
- volatile oil
- herb
- dutchmanspipe root
- ford dutchmanspipe
- ford
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B9/00—Essential oils; Perfumes
- C11B9/02—Recovery or refining of essential oils from raw materials
- C11B9/025—Recovery by solvent extraction
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/26—Aristolochiaceae (Birthwort family), e.g. heartleaf
- A61K36/264—Aristolochia (Dutchman's pipe)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B9/00—Essential oils; Perfumes
- C11B9/02—Recovery or refining of essential oils from raw materials
- C11B9/022—Refining
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B9/00—Essential oils; Perfumes
- C11B9/02—Recovery or refining of essential oils from raw materials
- C11B9/027—Recovery of volatiles by distillation or stripping
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/37—Extraction at elevated pressure or temperature, e.g. pressurized solvent extraction [PSE], supercritical carbon dioxide extraction or subcritical water extraction
Abstract
The invention discloses a kind of application of ford dutchmanspipe root or herb volatile oil in preparing anti-curing cancers and/antibacterial product.The step of ford dutchmanspipe root or herb volatile oil, extraction is:(1) ford dutchmanspipe root or herb is crushed, adds stirring progress steam distillation extraction after water, obtains water system containing volatile oil;(2) organic solvent is added into water system containing volatile oil to be extracted, discards the water layer separated, dehydration and drying agent dehydration is added into the organic solvent containing volatile oil, organic solvent is recovered by filtration to get to the pure volatile oil of drying.The ingredient of the ford dutchmanspipe root or herb obtained using steam distillation can have good antibacterial and active anticancer simultaneously.
Description
Technical field
The present invention relates to a kind of application of ford dutchmanspipe root or herb volatile oil in preparing anti-curing cancers and/antibacterial product.
Background technology
Volatile oil (volatile oils), also known as essential oil (essential oils) are one kind can volatilize at normal temperatures
, can with steam distillation, with the general name of the immiscible oily liquids of water.Most of volatile oil have aromatic odor.Volatilization
Oil is a kind of important active constituent, clinically in addition to directly applying the mainly crude drug containing volatile oil, can also apply and therefrom refine
Volatile oil, such as eucalyptus oil, peppermint oil.Volatile oil has diverging inducing diaphoresis, resuscitation with aromatics, regulating qi-flowing for relieving pain, dispelling wind and eliminating dampness, promoting blood circulation
In stagnation resolvation, dispelling cold temperature, clearing heat and detoxicating, relieving summer-heat dispel dirty, insecticidal and antibacterial the effects that.But the active constituent of the volatile oil of separate sources is not
Together, there is also larger differences for function, and such as peppermint oil dispelling-wind invigorating stomach, Angelica oil analgesia, radix bupleuri oil is brought down a fever, and American wormseed oil drives intestines
Worm etc..
Ford dutchmanspipe root or herb, belongs to Aristolochiaceae, ford dutchmanspipe root or herb platymiscium, the brave Tongcheng of alias five, centering grass, natural grass, root hyoscine,
The benefits of removing toxicity for detumescence, dispelling wind and eliminating pain, promoting the circulation of qi cough-relieving.It is distributed in Guangxi, Guangdong, Jiangxi, Zhejiang and the Fujian in China.Adaptability Yushan Hill
In paddy underbrush and in the stone gap of mountainous region.
Currently, the research for ford dutchmanspipe root or herb is less, decoction for oral administration is mostly used, or mashes or grinds and dress external application;Also not
About ford dutchmanspipe root or herb volatile oil extracting and the report used.
Invention content
In view of the deficiencies of the prior art, the object of the present invention is to provide a kind of ford dutchmanspipe root or herb volatile oil to prepare anti-curing cancers
With the application in/antibacterial product.
The step of the first aspect of the present invention, provides a kind of ford dutchmanspipe root or herb volatile oil, extraction be:
(1) ford dutchmanspipe root or herb is crushed, adds stirring progress steam distillation extraction after water, obtains water system containing volatile oil;
(2) organic solvent is added into water system containing volatile oil to be extracted, the water layer separated is discarded, to having containing volatile oil
Dehydration and drying agent dehydration is added in solvent, organic solvent is recovered by filtration to get to the pure volatile oil of drying.
Present invention discover that using steam distillation obtain ford dutchmanspipe root or herb ingredient can have simultaneously it is good it is antibacterial and
Active anticancer.
Preferably, in step (1), it is 50~100 mesh that ford dutchmanspipe root or herb, which is crushed to grain size,;It is further preferred that by ford dutchmanspipe root or herb
It is 60 mesh to be crushed to grain size.
Preferably, in step (1), the mass ratio that ford dutchmanspipe root or herb and water are added is 1:2~6, further preferably 1:3.
Preferably, in step (1), steam distillation extraction time is 3~6h.
Preferably, in step (2), the organic solvent is in ethyl acetate, ethyl acrylate, n-hexane or ether
It is one or more of;Still more preferably it is ether.
Preferably, in step (2), the dehydration and drying agent is selected from anhydrous sodium sulfate or dead plaster;It is further excellent
It is selected as anhydrous sodium sulfate.
Preferably, the chemical composition of ford dutchmanspipe root or herb volatile oil of the invention includes:Nopinene, pseudolimonene, limonene, γ-
Terpinene, terpinolene, linalool, alloocimene, trans- -2- menthols, camphor, terpinene-4-alcohol, alpha-terpineol, pepper
Ketone, δ-elemene, the long firpenes of α-, α-cubebene, β-patchoulene, australene, beta-elemene, comes into leaves at Bronyl acetate
Alkene, eneα-cedar, α-gurjunene, β-carypohyllene, β-farnesene, β-chamigrene, bicyclic big Mang bicyclogermacrene, β-Se Lin alkene, α-
Cloth lamb's-quarters alkene, γ-cadinene, δ-cadinene, the peaceful alkene of cubeb oil, 3,7 (11)-celery diene, α-epoxy cedrene, table blue gum
Alcohol, blue gum alkene, viridiflorol, guaiol, τ-cadinol, cedrol, new Andropogon intermedicus alcohol, cubeb oil enol, Wei get
Alcohol, α-cadinol, intermedeol, bisabol, iodothymol -1 (10)-alkene -9- alcohol, β-grapefruit alcohol, sesquialter myrtanol, α-cananga oil
Alkene -14- alcohol.
The second aspect of the present invention provides a kind of above-mentioned ford dutchmanspipe root or herb volatile oil and is preparing anti-curing cancers and/antibacterial product
In application.
In above application, the product includes:Food, drug or health products.
In above application, the bacterium is staphylococcus aureus, hay bacillus, Escherichia coli or Pseudomonas aeruginosa.Ford dutchmanspipe root or herb
Volatile oil has preferable fungistatic effect to staphylococcus aureus, hay bacillus, Escherichia coli and Pseudomonas aeruginosa.
In above application, the cancer is liver cancer or breast cancer.
The third aspect of the present invention, provides a kind of bacteriostatic agent, and active constituent is above-mentioned ford dutchmanspipe root or herb volatile oil.
The fourth aspect of the present invention provides a kind of pharmaceutical composition of anticancer, above-mentioned Tongcheng containing therapeutically effective amount
Brave volatile oil.
Terms used herein " therapeutically effective amount " indicate, treatment, the disease for improving targeting or illness or show can
The amount of therapeutic agent needed for the therapeutic effect of detection.
The pharmaceutical composition of the present invention is effective in comparatively wide dosage range.Actually take pharmaceutical composition of the present invention
For dosage can be determined according to related situation.These situations include:Physical condition, administration route, age, the body of subject
Again, to the individual reaction of drug, the severity etc. of symptom.
Preferably, various pharmaceutical dosage forms can be made in aforementioned pharmaceutical compositions, and corresponding medicine is matched according to different dosage forms
Oral suspension, capsule, emulsus agent is made in acceptable carrier on, is directly absorbed by stomach and is used for antitumaous effect;
Foaming agent, emulsus agent and suspension can be made, directly smear or be sprayed at lesions position, the principle of externally applied drug is to utilize volatilization
The osmosis of oil, by acting on lesion after skin or mucosa absorption, breast cancer is for example treated in externally applied drug.
Beneficial effects of the present invention are:
(1) present invention uses steam distillation and organic extraction techniques extraction purification to obtain ford dutchmanspipe root or herb volatile oil, institute for the first time
It states extraction process and is directed to ford dutchmanspipe root or herb volatile oil component feature, raw material coking can be effectively prevent, heating process is avoided to cause to volatilize
Thermal sensitivity in oil component, easily decompositions, oxidizable component damages avoid causing volatile oil loss of activity and volatilization oil quality reduction,
Improve the recovery rate of volatile oil component (including ingredient type and active component content);And method for extraction and purification is simple for process,
At low cost, distilled water and organic solvent in preparation process can reuse, the volatile oil obtained through preparation process of the present invention
Moisture content is low, and the main active constituent of volatile oil retains more comprehensively.
(2) present invention establishes composition and the quantitative analysis of ford dutchmanspipe root or herb volatile oil for the first time, passes through the spy of ford dutchmanspipe root or herb volatile oil
The difference of other species volatile oil active constituents of point, establishes the quality control detection side of ford dutchmanspipe root or herb Extraction Process of Volatile Oil respectively
The discriminating of method and ford dutchmanspipe root or herb volatile oil and discrimination method in ford dutchmanspipe root or herb.
(3) the ford dutchmanspipe root or herb volatile oil that extraction purification of the present invention obtains, experiment proves that having simultaneously good antibacterial and anti-
Cancer activity, has good bacteriostasis, to HepG2 to staphylococcus aureus, hay bacillus, Escherichia coli and Pseudomonas aeruginosa
(human liver cancer cell) and MCF-7 (human breast cancer cell) have good anticancer cancer resistant effect, are suitble to industrialized production and application.
Specific implementation mode
It is noted that described further below be all exemplary, it is intended to provide further instruction to the application.Unless another
It indicates, all technical and scientific terms used herein has usual with the application person of an ordinary skill in the technical field
The identical meanings of understanding.
It should be noted that term used herein above is merely to describe specific implementation mode, and be not intended to restricted root
According to the illustrative embodiments of the application.As used herein, unless the context clearly indicates otherwise, otherwise singulative
It is also intended to include plural form, additionally, it should be understood that, when in the present specification using term "comprising" and/or " packet
Include " when, indicate existing characteristics, step, operation, device, component and/or combination thereof.
It is less for the research of ford dutchmanspipe root or herb as background technology is introduced, decoction for oral administration is mostly used, or mash or grind
Mill dresses external application;At present also not about ford dutchmanspipe root or herb volatile oil extracting and the report used.Based on this, present applicant proposes one kind
Application of the ford dutchmanspipe root or herb volatile oil in preparing anti-curing cancers and/antibacterial product.
In a kind of embodiment of the application, a kind of ford dutchmanspipe root or herb volatile oil is provided, the step of extraction is:
(1) ford dutchmanspipe root or herb is crushed, adds stirring progress steam distillation extraction after water, obtains water system containing volatile oil;
(2) organic solvent is added into water system containing volatile oil to be extracted, the water layer separated is discarded, to having containing volatile oil
Dehydration and drying agent dehydration is added in solvent, organic solvent is recovered by filtration to get to the pure volatile oil of drying.
Present invention discover that using steam distillation obtain ford dutchmanspipe root or herb ingredient can have simultaneously it is good it is antibacterial and
Active anticancer.
The existing compound structure having found of plant that ford dutchmanspipe root or herb belongs to is mainly lignanoids, Phenylpropanoid Glycosides class, flavonoids, perfume (or spice)
Beans essence class, organic acid, alcohols, hydro carbons, saponins, amino acid and polysaccharide, volatile oil etc., and inventor has found ford dutchmanspipe root or herb volatilization
The characteristics of the characteristics of oil has oneself and classes of compounds (as shown in table 2), inventor's early-stage study ford dutchmanspipe root or herb volatile oil, using more
Kind mode attempts extraction ford dutchmanspipe root or herb volatile oil, and is detected by GC-MS, finds the volatile oil component overwhelming majority contained by ford dutchmanspipe root or herb
Closing object, solubility is extremely low in water, relatively preferably to be extracted by steam distillation, therefore proposes this programme, compares
For other volatile oil extracting modes, volatile oil extracting efficiency and volatile oil type ratio extract higher.
Wherein, during extraction by steam distillation ford dutchmanspipe root or herb volatile oil, raw material coking, inventor are easily led to the problem of
Experiment finds that the grain size of control ford dutchmanspipe root or herb dry powder is a vital factor, grain size is limited in 50 mesh or more, coking is existing
As obviously disappearing, on this basis, inventor further uses the ratio of suitable ford dutchmanspipe root or herb and distilled water, slows down raw material coke
Change problem and assisted extraction.
In specific embodiment, the mass ratio of the herein described ford dutchmanspipe root or herb cleaned after drying and distilled water is 1:2~
6, further preferably 1:3;It is preferably 60 mesh that the ford dutchmanspipe root or herb, which smashes grain size,;Inventor has found in experimental study, smashes
Grain is larger, and water vapour can be made to be flowed out from raw material gap, so that recovery rate reduces, and waste of resource;And it is too small to smash grain size,
Then so that water vapour can not uniformly penetrate raw material, causes steam to be escaped from the raw material washed open, it is not high to also result in recovery rate.
Further, since the extraction process of ford dutchmanspipe root or herb is newly-established, during the application, inventor is with reference to other plant
The extraction process of volatile oil, however it is different, and volatile oil of the present invention is contained more oxidizable ingredient and (is occupied with alkenes
It is more), to avoid heating process from causing thermal sensitivity in volatile oil component, easily decomposing, oxidizable component damages, water of the present invention steams
Steam distillation extraction time is limited in 3~6h;Suitable extraction time can ensure as far as possible by extracting volatile oil in ford dutchmanspipe root or herb
Component, while the wasting of resources caused by also avoiding extraction time long.
The organic solvent is selected from one or more of ethyl acetate, ethyl acrylate, n-hexane, ether, preferably
Ether;Inventor carries out experiment of single factor discovery for steam distillation-organic solvent extractionprocess, and ether is to ford dutchmanspipe root or herb volatile oil
Component extracting best results have better recovery rate compared to other organic solvents.
The dehydration and drying agent is selected from anhydrous sodium sulfate or dead plaster, preferably anhydrous sodium sulfate.
In the preferred embodiment of the application, the method for extraction and purification of ford dutchmanspipe root or herb volatile oil, including:
(1) it is 60 mesh to be smashed after taking fresh ford dutchmanspipe root or herb is clean to dry to grain size, and stirring progress water vapour after distilled water is added
Distillation, distillation extraction time are 5h, obtain water system containing volatile oil;Wherein, the matter for cleaning ford dutchmanspipe root or herb and distilled water after drying
Amount is than being 1:3;
(2) ether is added into water system containing volatile oil to be extracted, discards the water layer separated, it is molten to the ether containing volatile oil
Anhydrous sodium sulfate dehydration is added in liquid, ether is recovered by filtration and obtains dry pure volatile oil.
The main chemical compositions of the ford dutchmanspipe root or herb volatile oil of acquisition include β-chamigrene, β-carypohyllene, α-cloth lamb's-quarters alkene, β-pinane
Alkene, β-Se Lin alkene, limonene, linalool, bicyclogermacrene, longifolene.
More comprehensively, the main chemical compositions of ford dutchmanspipe root or herb volatile oil of the present invention include:Nopinene, pseudolimonene,
Limonene, γ-terpinene, terpinolene, linalool, alloocimene, trans- -2- menthols, camphor, terpinene-4-alcohol, α-pine
Oleyl alcohol, piperitone, Bronyl acetate, δ-elemene, the long firpenes of α-, α-cubebene, β-patchoulene, australene, β-olive
Fragrant alkene, longifolene, eneα-cedar, α-gurjunene, β-carypohyllene, β-farnesene, β-chamigrene, bicyclic big Mang bicyclogermacrene, β-
Plucked instrument woods alkene, α-cloth lamb's-quarters alkene, γ-cadinene, δ-cadinene, the peaceful alkene of cubeb oil, 3,7 (11)-celery diene, α-epoxy cypress
Alkene, table globulol, blue gum alkene, viridiflorol, guaiol, τ-cadinol, cedrol, new Andropogon intermedicus alcohol, cubeb oil
Enol, widdrol, α-cadinol, intermedeol, bisabol, iodothymol -1 (10)-alkene -9- alcohol, β-grapefruit alcohol, sesquialter Benny
Alcohol, α-muurolene -14- alcohol.
Process quality control, currently preferred reality are carried out for the ease of the more efficiently quality to ford dutchmanspipe root or herb volatile oil
The mode of applying is a kind of detection method by above-mentioned ford dutchmanspipe root or herb volatile oil, and the method includes using gas chromatography-mass spectrography (GC-
MS) analysis detection, the condition of detection are:HP-5MS chromatographic columns (30m × 0.25mm × 0.25 μm);Carrier gas:Helium;Flow velocity:
1.2mL/min, solvent delay 5min;Temperature program:Initial temperature is 60 DEG C, keeps 1min, 200 DEG C are warming up to 6 DEG C/min,
Keep 5min;280 DEG C are warming up to 5 DEG C/min, keeps 2min.Ionization mode:EI, 70eV;Ion source temperature:250℃;It carries
Gas:Helium;Column flow rate:1.2mL/min;
Detect and control the ford dutchmanspipe root or herb chemical composition of volatile oil, including β-chamigrene, β-carypohyllene, α-cloth lamb's-quarters alkene, β-
The control of the ingredients such as firpene, β-Se Lin alkene, limonene, linalool, bicyclogermacrene, longifolene;
More comprehensively, the main chemical compositions for detecting the ford dutchmanspipe root or herb volatile oil include:Nopinene, pseudolimonene, lemon
Lemon alkene, γ-terpinene, terpinolene, linalool, alloocimene, trans- -2- menthols, camphor, terpinene-4-alcohol, α-pine tar
Alcohol, piperitone, Bronyl acetate, δ-elemene, the long firpenes of α-, α-cubebene, β-patchoulene, australene, β-elemi
Alkene, longifolene, eneα-cedar, α-gurjunene, β-carypohyllene, β-farnesene, β-chamigrene, bicyclic big Mang bicyclogermacrene, β-plucked instrument
Woods alkene, α-cloth lamb's-quarters alkene, γ-cadinene, δ-cadinene, the peaceful alkene of cubeb oil, 3,7 (11)-celery diene, α-epoxy cedrene,
Table globulol, blue gum alkene, viridiflorol, guaiol, τ-cadinol, cedrol, new Andropogon intermedicus alcohol, cubebene
Alcohol, widdrol, α-cadinol, intermedeol, bisabol, iodothymol -1 (10)-alkene -9- alcohol, β-grapefruit alcohol, sesquialter myrtanol,
α-muurolene -14- alcohol.
Above-mentioned detection method is used for the discriminating of ford dutchmanspipe root or herb volatile oil in the embodiment of the application.
For example, a kind of discrimination method of ford dutchmanspipe root or herb volatile oil, the method includes volatile oil to be measured is used gas-chromatography-
Mass spectrography (GC-MS) analysis detection, the condition of detection are:HP-5MS chromatographic columns (30m × 0.25mm × 0.25 μm);Carrier gas:Helium
Gas;Flow velocity:1.2mL/min, solvent delay 5min;Temperature program:Initial temperature is 60 DEG C, keeps 1min, is heated up with 6 DEG C/min
To 200 DEG C, 5min is kept;280 DEG C are warming up to 5 DEG C/min, keeps 2min.Ionization mode:EI, 70eV;Ion source temperature:
250℃;Carrier gas:Helium;Column flow rate:1.2mL/min;
Volatile oil testing result to be measured and ford dutchmanspipe root or herb chemical composition of volatile oil (β-chamigrene, β-carypohyllene, α-cloth lamb's-quarters alkene,
Nopinene, β-Se Lin alkene, limonene, linalool, bicyclogermacrene, longifolene) it compares.
A kind of discrimination method of ford dutchmanspipe root or herb, the method includes the extractions of species volatile oil to be measured and species volatile oil to be measured
Detection,
The extraction of the species volatile oil to be measured includes:(1) take species to be measured clean dry after smash to grain size be 50~
100 mesh are added stirring progress steam distillation after distilled water, obtain water system containing volatile oil;
(2) organic solvent is added into water system containing volatile oil to be extracted, the water layer separated is discarded, to having containing volatile oil
Dehydration and drying agent dehydration is added in solvent, organic solvent is recovered by filtration and obtains dry pure volatile oil;
The detection of species volatile oil to be measured includes:By species volatile oil to be measured using gas chromatography-mass spectrography (GC-MS) point
Analysis detection, the condition of detection are:HP-5MS chromatographic columns (30m × 0.25mm × 0.25 μm);Carrier gas:Helium;Flow velocity:1.2mL/
Min, solvent delay 5min;Temperature program:Initial temperature is 60 DEG C, keeps 1min, is warming up to 200 DEG C with 6 DEG C/min, keeps
5min;280 DEG C are warming up to 5 DEG C/min, keeps 2min.Ionization mode:EI, 70eV;Ion source temperature:250℃;Carrier gas:
Helium;Column flow rate:1.2mL/min;
Volatile oil testing result to be measured and ford dutchmanspipe root or herb chemical composition of volatile oil (β-chamigrene, β-carypohyllene, α-cloth lamb's-quarters alkene,
Nopinene, β-Se Lin alkene, limonene, linalool, bicyclogermacrene, longifolene) it compares.
In addition, the present invention has studied the possible function of series of the ford dutchmanspipe root or herb volatile oil, above-mentioned ford dutchmanspipe root or herb volatilization is disclosed
The application of oil.
Inventor also studies the function of excessive ESSENTIAL OIL early period, such as carves dish volatile oil, bean curd dish volatile oil, seta
Wild chervil volatile oil, Geophila herbacea (L) O Kumtze volatile oil etc., but different phytovolatilization oil component is sufficiently complex, and source is different, function there is also
Larger difference understands according to inventor, while having antibacterial, antitumor activity multifunctional usage volatile oil type still less,
Function is still mainly related with active constituent and its related ratio, and inventor is found that bean curd dish volatile oil has the function of this early period,
Seta wild chervil volatile oil also has effects that this, and during research ford dutchmanspipe root or herb volatile oil is crossed, inventor has found that the volatile oil is same
When there are good antibacterial and active anticancer multiple functions, to staphylococcus aureus, hay bacillus, Escherichia coli and green pus bar
Bacterium has good bacteriostasis, presses down with good anticancer to HepG2 (human liver cancer cell) and MCF-7 (human breast cancer cell)
Cancer effect.
In the other embodiment of the application, discloses a kind of above-mentioned ford dutchmanspipe root or herb volatile oil and preparing anti-curing cancers
With the application in/antibacterial product.
Further, the product includes:Food, drug or health products.The bacterium is staphylococcus aureus, withered grass bar
Bacterium, Escherichia coli or Pseudomonas aeruginosa.Ford dutchmanspipe root or herb volatile oil is to staphylococcus aureus, hay bacillus, Escherichia coli and green pus bar
Bacterium has preferable fungistatic effect.The cancer is liver cancer or breast cancer.
Such as drug, it is corresponding pharmaceutically acceptable according to different dosage form apolegamys using ford dutchmanspipe root or herb volatile oil as raw material
Oral suspension, capsule, emulsus agent is made in carrier, is directly absorbed by stomach and is used for antitumaous effect;Foam can be made
Agent, emulsus agent and suspension, directly smear or are sprayed at lesions position, and the principle of externally applied drug is made using the infiltration of volatile oil
With by acting on lesion after skin or mucosa absorption, breast cancer is for example treated in externally applied drug.
In order to enable those skilled in the art can clearly understand the technical solution of the application, below with reference to tool
The technical solution of the application is described in detail in the embodiment of body.
Test material used in following embodiment is the test material of this field routine, can be purchased by commercial channel
It can buy.
Embodiment 1:Preliminary experiment
Different extracting methods is respectively adopted to extract ford dutchmanspipe root or herb volatile oil, it is specific as follows:
1. steam distillation:
(1) it takes the dry ford dutchmanspipe root or herbs of 500g to smash, stirring progress steam distillation after 1500g distilled water is added, when distilling extraction
Between be 5h, obtain water system containing volatile oil;
(2) ether is added into water system containing volatile oil to be extracted, discards the water layer separated, it is molten to the ether containing volatile oil
Anhydrous sodium sulfate dehydration is added in liquid, ether is recovered by filtration and obtains dry pure volatile oil.
2. Simultaneous distillation:
It weighs the dry ford dutchmanspipe root or herbs of 500g to smash in distilling flask towel, 1500g aquae destillatas is added, while measuring ether and being burnt in distillation
In bottle, moral extraction equipment is connected while being steamed, while steaming and extraction is stayed to collect ether solution, anhydrous sodium sulfate is used in combination to be dehydrated, ether extraction
Liquid is concentrated in rotavapor under vacuum, obtains volatile oil.
3. aether backflow extraction:
It weighs the dry ford dutchmanspipe root or herbs of 500g to smash in distilling flask towel, ether is added and carries out reflux extraction.
The investigation of preliminary experiment the results are shown in Table 1.
1 preliminary result of table
As can be seen from Table 1, higher using the volatile oil yield of steam distillation, therefore, for waving in subsequent experimental
The extraction of hair oil uses steam distillation.
Embodiment 2:The method for extraction and purification of ford dutchmanspipe root or herb volatile oil
(1) take the fresh ford dutchmanspipe root or herbs of 500g clean dry after smash to grain size be 60 mesh, be added 1500g distilled water after stir into
Row steam distillation, distillation extraction time are 5h, obtain water system containing volatile oil;
(2) ether is added into water system containing volatile oil to be extracted, discards the water layer separated, it is molten to the ether containing volatile oil
Anhydrous sodium sulfate dehydration is added in liquid, ether is recovered by filtration and obtains dry pure volatile oil;Recovery rate is 0.32%, volatilization
Moisture content in oil is 0.55%.
Embodiment 3:The detection method of ford dutchmanspipe root or herb volatile oil
The ford dutchmanspipe root or herb volatile oil that embodiment 2 obtains is detected using GC-MS, the condition specifically detected is as follows:HP-
5MS chromatographic columns (30m × 0.25mm × 0.25 μm);Carrier gas:Helium;Flow velocity:1.2mL/min, solvent delay 5min;Heat up journey
Sequence:Initial temperature is 60 DEG C, keeps 1min, is warming up to 200 DEG C with 6 DEG C/min, keeps 5min;It is warming up to 280 with 5 DEG C/min
DEG C, keep 2min.Ionization mode:EI, 70eV;Ion source temperature:250℃;Carrier gas:Helium;Column flow rate:1.2mL/min.Altogether
Detect that 51 kinds of chemical composition in ford dutchmanspipe root or herb volatile oil, specific chemical composition and relative amount are shown in Table 2.
2 ford dutchmanspipe root or herb chemical composition of volatile oil of table
Embodiment 4:The bacteriostasis of ford dutchmanspipe root or herb volatile oil
1. testing strain and culture medium
Test strain:Four kinds of bacterium bacterial strains of selection are used for bacteriostatic test, including gram-positive bacteria:Staphylococcus aureus
(ATCC 6538), bacillus subtilis (ATCC 6633) and Gram-negative bacteria:Escherichia coli (ATCC 25922), green pus bar
Bacterium (ATCC 27853).
Experiment culture medium:Bacterium MHB (Mueller Hinton Broth) culture medium.
2. test method
(1) it sterilizes:
It is damp and hot at 121 DEG C that required culture medium, test tube, culture dish, filter paper (6mm), pipette tips, distilled water etc. will be tested
Sterilize 20min, and bacterium of having gone out is placed superclean bench ultraviolet sterilization 30min.
(2) preparation of bacteria suspension:
It is inoculated in corresponding MHB culture mediums from one ring thalline of picking on the thalline inclined-plane through overactivation, is put into constant temperature and shakes
Culture overnight under the conditions of 37 DEG C in bed.Draw respectively it is above it is cultured in exponential phase for trying bacterium solution 0.5mL, use MH
Bacteria suspension of the broth dilution at 0.5 Maxwell than turbid standard.After above-mentioned bacteria suspension is carried out 1: 100 dilution with MH meat soups again, 1 is obtained
~2 × 106CFU/ml is spare.
(3) for reagent object:
Control drug:Chloramphenicol;Sample:The 5mg/mL dilutions of 2 gained ford dutchmanspipe root or herb volatile oil of embodiment.
(4) measurement of bacteriostatic activity:
Bacteriostatic activity is measured using filter paper enzyme.Under aseptic condition, MHA culture mediums are poured into sterilized culture dish
Solid plate is made after cooled and solidified in 20mL.The bacteria suspension that 100 μ L have diluted is added into each solid plate, coating is equal
It is even, 5min is placed, culture medium is made to fully absorb bacterium solution.With marking pen on having bacterio-agar tablet label.It will be gone out with aseptic nipper
The filter paper of the drying of complete bacterium presss from both sides out, is put into the culture dish for writing corresponding control number, is dripped to the filter paper of sterilized drying
Add 10 μ L of prepared sample solution or chloromycetin solution, each sample do three it is parallel, be put into 37 DEG C of constant incubator cultures
25h.Incubation time takes out culture dish after arriving and observes transparent inhibition zone, and each inhibition zone is measured using crossing method micrometer
The diameter of two vertical direction, it is measurement result to take its average value (mm), each bacteriostatic experiment is parallel to be repeated 3 times.With
The size of antibacterial circle diameter carries out the judgement of bacteriostatic activity power, and inhibition zone shows bigger, then show its antibacterial activity
It is stronger.
(5) measurement of MIC:
It draws the bacterium solution diluted to be added in 96 porocyte culture plates, per 100 μ L of hole, and it is dense that configured good difference is added
The 100 μ L of drug of degree.Hole second from the bottom is not added with drug (only plus culture medium and bacterium) and dilution 100 μ L of bacterium solution is added, and is given birth to for bacterium
Long control wells.Last is classified as positive control chloramphenicol.It is placed on oscillator and vibrates 1min, make after solution mixes well in hole,
The oscillation of 96 orifice plates is shaken up after being placed on and being cultivated 16~18 hours in 37 DEG C of constant incubators, into every hole, addition sterile water is matched
The 20 μ L of 1%TTC (2,3, 5-Triphenyltertrazoliumchloride) solution set, oscillation shake up in 37 DEG C of constant incubators culture 30~
It 60 minutes, observes per hole color change.It takes and volatile oil contained by last hole of color change is not presented a concentration of its is minimum antibacterial
Concentration MIC.
The measurement for repeating all MIC values is triplicate.
3. experimental result
Fungistatic effect indicates that measurement result is shown in Table 3 with minimal inhibitory concentration MIC.
3 ford dutchmanspipe root or herb volatile oil bacteriostatic activity of table
As can be seen from Table 3,4 kinds of ford dutchmanspipe root or herb volatile oil pair has certain inhibiting effect for examination bacterium, to golden yellow Portugal
The minimum inhibitory concentration of grape coccus is 0.078mg/mL, and the minimum inhibitory concentration to hay bacillus is 0.078mg/mL, to large intestine
The minimum inhibitory concentration of bacillus is more than 5.00mg/mL, and the minimum inhibitory concentration to Pseudomonas aeruginosa is 2.50mg/mL.
By be embodied example illustrate ford dutchmanspipe root or herb volatile oil can be used as natural antibacterial agent for food, drugs and health care products or
Cosmetic industry, instead of the synthetic antibacterial agents of toxic side effect.
Embodiment 5:The cytotoxicity of ford dutchmanspipe root or herb volatile oil
1. cell culture
HepG2 (human liver cancer cell) and MCF-7 (human breast cancer cell) cell strain, which are used, contains 10% newborn bovine serum and blueness
The RPMI1640 culture mediums of each 100U/mL of mycin, streptomysin be placed in 37 DEG C, saturated humidity, 5%CO2Incubator in cultivate.
2.MTT methods detect cell viability
The tumour cell of logarithmic growth phase is suspended in the RPMI-1640 containing 10% fetal calf serum and 2mM glutamine
In culture medium, single cell suspension is gently blown and beaten into glass pipette, is counted with TC10 automated cell calculating instruments.By cell inoculation in
96 well culture plates, cell density are 5 × 103A/mL, per 200 μ L of hole.In 37 DEG C, 5%CO2Incubator preculture 25h make carefully
After born of the same parents' adherency, it is respectively 0.016,0.031,0.063,0.125,0.250,0.500,1 and 2mg/ that the concentration prepared in advance, which is added,
The volatile oil of mL (is prepared) solution by embodiment 2 and handled tumour cell.And set negative control and blank control.Distinguish again
Continuous culture 25h, 48h, 72h, are then measured with mtt assay.The MTT solution of 10 μ L 5mg/mL is added per hole, continues to cultivate 2h
Afterwards, supernatant is sucked.100 μ L DMSO are added per hole, setting micro oscillator oscillation 10min makes crystallization be completely dissolved, in
Multiskan Ascent microplate reader measures absorbance (A) value at wavelength 570nm, calculates cell proliferation inhibition rate, the measurement
In triplicate.Cell proliferation inhibition rate calculates as follows.
Cell proliferation inhibition rate/%=[1-ODProcessing group/ODControl]× 100%
3. experimental result
Required sample concentration i.e. (IC when cytotoxicity is to inhibit 50% cell growth50Value) it indicates, measurement result is shown in Table 4.
4 ford dutchmanspipe root or herb volatile oil of table is to HepG2, MCF-7 cytotoxic activity
As can be seen from Table 4, ford dutchmanspipe root or herb volatile oil has certain inhibiting effect to two kinds of cells of HepG2, MCF-7,
To 72 hours IC of HepG250For 0.69mg/mL;To 72 hours IC of MCF-750For 0.22mg/mL.
The above results illustrate that ford dutchmanspipe root or herb volatile oil can be used as natural anti-cancer cell agent for food, drug or health care conduct
Industry.
The foregoing is merely the preferred embodiments of the application, are not intended to limit this application, for the skill of this field
For art personnel, the application can have various modifications and variations.Within the spirit and principles of this application, any made by repair
Change, equivalent replacement, improvement etc., should be included within the protection domain of the application.
Claims (10)
1. a kind of ford dutchmanspipe root or herb volatile oil, characterized in that the step of extraction is:
(1) ford dutchmanspipe root or herb is crushed, adds stirring progress steam distillation extraction after water, obtains water system containing volatile oil;
(2) organic solvent is added into water system containing volatile oil to be extracted, the water layer separated is discarded, to organic molten containing volatile oil
Dehydration and drying agent dehydration is added in agent, organic solvent is recovered by filtration to get to the pure volatile oil of drying.
2. ford dutchmanspipe root or herb volatile oil as described in claim 1, characterized in that in step (1), it is 50 that ford dutchmanspipe root or herb, which is crushed to grain size,
~100 mesh;It is further preferred that it is 60 mesh that ford dutchmanspipe root or herb, which is crushed to grain size,;
Or, in step (1), the mass ratio that ford dutchmanspipe root or herb and water are added is 1:2~6, further preferably 1:3.
3. ford dutchmanspipe root or herb volatile oil as described in claim 1, characterized in that in step (1), steam distillation extraction time is 3
~6h.
4. ford dutchmanspipe root or herb volatile oil as described in claim 1, characterized in that in step (2), the organic solvent is selected from acetic acid second
One or more of ester, ethyl acrylate, n-hexane or ether;Further preferably ether.
5. ford dutchmanspipe root or herb volatile oil as described in claim 1, characterized in that in step (2), the dehydration and drying agent is selected from anhydrous
Sodium sulphate or dead plaster;Still more preferably it is anhydrous sodium sulfate.
6. ford dutchmanspipe root or herb volatile oil as described in claim 1, characterized in that the chemical composition of ford dutchmanspipe root or herb volatile oil includes:β-pinane
Alkene, pseudolimonene, limonene, γ-terpinene, terpinolene, linalool, alloocimene, trans- -2- menthols, camphor, pine tar
Alkene -4- alcohol, alpha-terpineol, piperitone, Bronyl acetate, δ-elemene, the long firpenes of α -, α-cubebene, β-patchoulene,
It is australene, beta-elemene, longifolene, eneα-cedar, α-gurjunene, β-carypohyllene, β-farnesene, β-chamigrene, bicyclic big
Mang bicyclogermacrene, β-Se Lin alkene, α-cloth lamb's-quarters alkene, γ-cadinene, δ-cadinene, the peaceful alkene of cubeb oil, 3,7 (11)-celery diene,
α-epoxy cedrene, table globulol, blue gum alkene, viridiflorol, guaiol, τ-cadinol, cedrol, new Andropogon intermedicus
Alcohol, cubeb oil enol, widdrol, α-cadinol, intermedeol, bisabol, iodothymol -1 (10)-alkene -9- alcohol, β-grapefruit
Alcohol, sesquialter myrtanol, α-muurolene -14- alcohol.
7. a kind of claim 1~6 any ford dutchmanspipe root or herb volatile oil is preparing anti-curing cancers and answering in/antibacterial product
With.
8. the use as claimed in claim 7, characterized in that the bacterium is staphylococcus aureus, hay bacillus, Escherichia coli
Or Pseudomonas aeruginosa.Ford dutchmanspipe root or herb volatile oil has staphylococcus aureus, hay bacillus, Escherichia coli and Pseudomonas aeruginosa preferable
Fungistatic effect;
Or, the cancer is liver cancer or breast cancer.
9. a kind of bacteriostatic agent, characterized in that active constituent is any ford dutchmanspipe root or herb volatile oil of claim 1~6.
10. a kind of pharmaceutical composition of anticancer, characterized in that it is any described that it contains the claim 1~6 of therapeutically effective amount
Ford dutchmanspipe root or herb volatile oil.
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| CN110227071B (en) * | 2019-08-05 | 2019-11-22 | 江西省林业科学院 | Compound borneol antibacterial agent |
| CN113025418A (en) * | 2021-03-16 | 2021-06-25 | 山东智汇达海洋生物科技有限公司 | Preparation and application of volatile oil of Odontia papyrifera |
| CN113025418B (en) * | 2021-03-16 | 2022-12-06 | 山东智汇达海洋生物科技有限公司 | Preparation and application of volatile oil of Odontia papyrifera |
| CN113021558A (en) * | 2021-03-17 | 2021-06-25 | 浙江省林业科学研究院 | Preparation method of wear-resistant antibacterial bamboo straw |
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