CN107163013A - A kind of method that Osthole is prepared from plant and the application in cosmetics are prepared - Google Patents

Abstract

The invention discloses a kind of method that Osthole is prepared from plant and the application in cosmetics are prepared.The present invention is first pre-processed to the fruit position containing Osthole, then by steps such as ethanol extraction, melt cinder separation, the alcohol crystal of Osthole runic precipitating, accurate concentration and recrystallizations, obtains Osthole monomer reactivity composition.The method that the present invention is provided not only can effectively dispel impurity, and the rate of transform of Osthole is high.The composition of acquisition has the functions such as antipruritic antibacterial anti-inflammatory, desinsection, antiallergy, spasmolysis decompression, anti-arrhythmia, enhancing immunologic function, emulsification system, surfactant system can be prepared into, as antiallergy, antipruritic desinsection, antibacterial anti-inflammatory, go acne the effects such as cosmetic skincare product.

Description

A kind of method that Osthole is prepared from plant and the application in cosmetics are prepared

Technical field

The invention belongs to medicine and cosmetic field, more particularly to a kind of method that Osthole is prepared from plant and Prepare the application in cosmetics.

Background technology

Osthole also known as osthole, or osthole, or osthol (Osthole, Ost), are primarily present It is samphire cnidium monnieri Cnidium monnieri (L.) Cuss. or Cnidium in Umbelliferae and rutaceae Main active in formosanum YABE dry mature fruit, its chemical name is 7- methoxyl group -8- isopentene groups Cumarin (7-methoxy-8- [3-methylpent-2-eny1] coumarin), is a kind of hydrocarbon of content highest in frutus cnidii Butylcoumariii class active chemical, is colourless column crystallization, molecular formula:C₁₅H₁₆O₃, relative molecular mass is 244.29, fusing point For 82~84 DEG C.The pharmacological experiment study of Osthole shows：Osthole have anti-hypertension, anti-arrhythmia, anti-aging, The effect such as antitumor, anti-osteoporosis disease, anti-inflammatory and resistance state, enhancing immunologic function.In terms of daily use chemicals external application, frutus cnidii have Anti-allergy action, has stronger antihistamine effect, energy obvious antagonizing histamine, chronic reactive material have stronger antiallergy Effect；There is preferably treatment and health-care effect to various allergy, pruitus, psoriasis, zona, eczema；This Outside, frutus cnidii have stronger resisting pathogenic microbes effect, to acrothesium floccosum, gypsum sample microspore bacterium, ulotrichy budlet Born of the same parents bacterium and broad spectrum of bacteria have inhibitory action, there is stronger killing action to Trichomonas vaginalis, can suppress the formation of various inflammatory factors, Antipruritic, the function of removing acne with antibacterial anti-inflammatory well, desinsection.

In order to adapt to Osthole it is growing the need for, the preparation research of Osthole has faster development, but Have clearly disadvantageous.Osthole and different fennel are prepared from frutus cnidii as patent of invention CN201110151743.5 discloses one kind The process of sedanonic acid lactone；Method is Common Cnidium Fruit first through supercritical CO₂Extraction, obtains common cnidium fruit P.E, then through poly- Amide resin is isolated and purified, and obtains crude product, is most further purified through high speed adverse current chromatogram afterwards, obtains Osthole and isoimpinellin is pure Product；The process equipment is expensive, production cost is high, is not suitable for carrying out large-scale industrial production.And for example patent of invention CN201010182729.7 is disclosed in a kind of " technique for extracting Osthole " method, the process and is employed micro-filtration, surpasses Filter, the crystallization of nanofiltration petroleum ether, the method for absolute ethyl alcohol repeated recrystallize, the process is not only relative complex, cost is high, and Petroleum ether is also easy to produce potential safety hazard for big industrial production.And for example patent of invention CN 201010531931.6 discloses " a kind of again The highly basic such as NaOH, KOH have been used to handle in the preparation method of Osthole ", the process, its Osthole of highly basic malleable Structure produce side reaction.Also and for example patent of invention CN 201010527673.4 discloses " one kind extraction from Cnidium Monnieri The method of Osthole ", though the process method is simple, also is adapted for large-scale industrial production；But used in process The ethanol solution that concentration is more than 90% is crystallized and recrystallized, because 90% ethanol solution is to Osthole and impurity Dissolubility is all very strong, so, not only need recrystallization often, and the influence to the yield of product is also larger.The present invention Above-mentioned deficiency can be overcome by extracting the method for separation Osthole, be produced using this method, high income is simple to operate, and energy consumption is low, easily Realize that industrialization is amplified, yet there are no and have been reported that.

The content of the invention

The primary and foremost purpose of the present invention is to overcome the shortcoming and deficiency of prior art to prepare cnidium monnieri from plant there is provided one kind The method of sub- element.

Another object of the present invention is to provide the Osthole prepared by the above method.

It is still another object of the present invention to provide the application of the above method and Osthole in cosmetics are prepared.

The purpose of the present invention is achieved through the following technical solutions：A kind of method that Osthole is prepared from plant, including Following steps：

(1) extraction of Osthole：Using the fruit of samphire as raw material, cleaned, dried, crushed, obtain thick Powder；With the ethanol solution heating extraction or heating temperature leaching ultrasonic extraction of percent by volume 65%~95%；After having extracted, solid-liquid point From obtaining the extract solution containing Osthole；

(2) separation of Osthole runic：The extract solution containing Osthole is taken, ethanol is reclaimed to complete, obtains concentrate； Add temperature in concentrate to be dissolved for 40~80 DEG C of hot water, then cool down, then at 1~25 DEG C of standing 24 hours with On, separation of solid and liquid discards liquid, collects solid, obtains the crude extract containing Osthole；

(3) polishing purification of Osthole monomer component：By step 1. or 1. and 2. step carries out polishing purification；

1. crystallize for the first time：Concentration is added in Osthole crude extract water-soluble for the ethanol of percent by volume 50~60% Liquid, is dissolved by heating, and is cooled to room temperature, is stood for the first time, separation of solid and liquid takes liquid, discards solid；Liquid obtained by separation of solid and liquid in Second of standing is fully separated out to crystal in 0~10 DEG C of environment, then separation of solid and liquid, discards liquid, collects crystallization；

2. recrystallize；1. obtained crystallization is recrystallized according still further to step, the polishing purification crystallization of Osthole is obtained.

Samphire described in step (1) be preferably cnidium monnieri Cnidium monnieric (L.) Cusson. or Cnidium formosanum YABE。

Coarse powder described in step (1) is preferably the coarse powder of 15~20 mesh.

Ethanol solution preferred concentration described in step (1) is the ethanol solution of percent by volume 85~90%.

The number of times of heating extraction described in step (1) is preferably 2~3 times；Specific steps are preferably as follows：Extraction time is At 2 times：The ethanol solution equivalent to 10 times of volumes (L) of meal quality (kg) is added for the first time, is sufficiently stirred for mixing, in 70~ 100 DEG C of refluxing extractions 1~3 hour；The ethanol solution equivalent to 8 times of volumes of meal quality is added for the second time, in 70~100 DEG C Refluxing extraction 0.5~2 hour；When extraction time is 3 times：It is preceding twice with extraction time be 2 times when the step of, third time add phase When in the ethanol solution of 8 times of volumes of meal quality, in 70~100 DEG C of refluxing extractions 0.5~2 hour；After having extracted, solid-liquid point From 2~3 isolated liquid of merging obtain the extract solution containing Osthole；Specific steps are more preferably as follows：Extract secondary When number is 2 times：The ethanol solution equivalent to 10 times of volumes (L) of meal quality (kg) is added for the first time, is sufficiently stirred for mixing, in 80 DEG C of refluxing extractions 2 hours；The ethanol solution equivalent to 8 times of volumes of meal quality is added for the second time, in 80 DEG C of refluxing extractions 1.5 hour；When extraction time is 3 times：It is preceding twice with extraction time be 2 times when the step of, third time add equivalent to coarse powder matter The ethanol solution of 8 times of volumes is measured, in 80 DEG C of refluxing extractions 1.5 hours；After having extracted, separation of solid and liquid merges 2~3 separation Obtained liquid, obtains the extract solution containing Osthole.

The ultrasound condition in heating temperature leaching ultrasonic extraction described in step (1) is preferably 200~300Hz；More preferably 250Hz。

The number of times of heating temperature leaching ultrasonic extraction described in step (1) is preferably 2~3 times；Specific steps are preferably as follows：Carry Take number of times be 2 times when：The ethanol solution equivalent to 10 times of volumes (L) of meal quality (kg) is added for the first time, is sufficiently stirred for mixing It is even, soak ultrasonic extraction 0.5~2 hour in 60~80 DEG C of temperature；Second of addition is molten equivalent to the ethanol of 8 times of volumes of meal quality Liquid, ultrasonic extraction is soaked 0.5~1.5 hour in 60~80 DEG C of temperature；When extraction time is 3 times：It is preceding twice with extraction time be 2 times when The step of, third time adds the ethanol solution equivalent to 8 times of volumes of meal quality, in 60~80 DEG C of temperature leaching ultrasonic extractions 0.5 ~1.5 hours；After having extracted, separation of solid and liquid merges 2~3 isolated liquid, obtains the extract solution containing Osthole； Specific steps are more preferably as follows：When extraction time is 2 times：Add for the first time equivalent to 10 times of volumes (L) of meal quality (kg) Ethanol solution, be sufficiently stirred for mixing, in 60 DEG C of temperature leaching ultrasonic extractions 1.5 hours；Second adds equivalent to 8 times of meal quality The ethanol solution of volume, ultrasonic extraction is soaked 1 hour in 60 DEG C of temperature；When extraction time is 3 times：Preceding is twice 2 with extraction time The step of when secondary, third time adds the ethanol solution equivalent to 8 times of volumes of meal quality, small in 60 DEG C of temperature leaching ultrasonic extractions 1 When；After having extracted, separation of solid and liquid merges 2~3 isolated liquid, obtains the extract solution containing Osthole.

Hot water preferable temperature described in step (2) is 50~70 DEG C of hot water；More preferably 60 DEG C of hot water.

The addition of hot water described in step (2) is preferably by carrying that 1kg raw materials (fruit of samphire) are obtained The volume that concentrate is added after hot water is taken to be calculated for 1~2L；The volume more preferably added after hot water calculates for 1.5~1.8L.

Cooling described in step (2) is preferably natural cooling.

The degree of cooling described in step (2) is preferably to be cooled to less than 30 DEG C；More preferably it is cooled to less than 25 DEG C.

The condition of standing described in step (2) is preferably to stand 24~30h in 2~8 DEG C；More preferably 3~6 DEG C standings 26~28h.

Step (3) 1. described in the concentration of ethanol water be preferably percent by volume 53~57%；More preferably body Product percentage 55~56%.

Step (3) 1. described in ethanol water addition be preferably press 1kg raw materials (fruit of samphire) Obtained crude extract matches 1~3L ethanol waters and calculated；More preferably proportioning 2L ethanol waters are calculated.

Step (3) 1. described in room temperature refer to 5~35 DEG C；Preferably 10~30 DEG C；More preferably 20~28 DEG C；Most Preferably 25 DEG C.

Step (3) 1. described in first time stand time be preferably 7~8 hours.

Step (3) 1. described in second standing time be preferably more than 48 hours；More preferably 72 hours.

Step (3) 1. described in second standing temperature be preferably 0~8 DEG C；More preferably 1~2 DEG C.

Step (3) 2. described in the number of times of recrystallization be preferably 1 time, the polishing purification crystallization of obtained Osthole Purity content >=98% (w/w).

Heretofore described separation of solid and liquid can be separated by centrifugation or separated by filter type.

Application of the described method in cosmetics are prepared.

A kind of refined Osthole, is prepared by the above method, purity content >=98% (w/w).

Application of the described refined Osthole in cosmetics are prepared.

Described cosmetics be with antiallergy, antibacterial anti-inflammatory, function of removing acne cosmetics.

Antiallergy refers to that cosmetics prepared by Osthole can suppress the release of histamine, can suppress passive skin mistake strongly Quick reaction, can substantially suppress delayed type hypersensitivity, DTH, with antiallergic action, itching-relieving action；Antibacterial anti-inflammatory, acne are gone to refer to snake Cosmetics prepared by machine tool element have stronger antibacterial and anti-inflammation functions, can suppress staphylococcus aureus, staphylococcus aureus resistance to The bacteriums such as medicine bacterium, Escherichia coli, shigella dysenteriae, salmonella typhimurium, Pseudomonas aeruginosa；Can significantly inhibit it is various it is acute and Chronic inflammation, with antibacterial anti-inflammatory well, function of removing acne.

Application of the described refined Osthole in cosmetics are prepared, comprises the following steps：By described frutus cnidii Element is first dissolved or dissolved by heating in preparing in the drug excipient or carrier of cosmetics permission, then is mixed with other compositions.

Content of the described refined Osthole in described cosmetics is therapeutically effective amount.

The described drug excipient or carrier that prepare cosmetics permission can be selected from：It is solvent, solubilizer, preservative, anti- Oxygen agent, pH adjusting agent, penetrating agent, liposome, NMF, thickener, chelating agent, skin sense conditioning agent, surfactant, emulsification Agent, propulsion/propellant, essence, pigment, and other functional additives.

The form of described cosmetics is emulsification system, surfactant system, externally-applied liniment, ointment.

In order to illustrate the beneficial effect that Osthole and its application in cosmetics are prepared from plant, the present invention is with gold Staphylococcus aureus, staphylococcus aureus resistance bacterium, Escherichia coli, shigella dysenteriae, salmonella typhimurium, green pus bar Bacterium is research object, and it is notable to prove that the Osthole of the preparation of the present invention has by quick paper disk method, doubling dilution experiment Antibacterial and anti-inflammation functions；Using male mice as research object, scorching method is caused to test by mouse auricular concha, it was demonstrated that frutus cnidii of the invention Element has the effect of significant anti-inflammatory analgetic；Using mouse and rat as research object, by suppressing mouse models of passive skin irritability (PCA) Reaction, it was demonstrated that Osthole of the invention has significant anti-allergic effects；

The present invention has the following advantages and effect relative to prior art：

(1) method that the present invention prepares Osthole from plant frutus cnidii, overcomes there is now for Osthole preparation CO₂Supercritical fluid extraction, acid-alkali treatment method, petroleum ether processing crystallisation, high concentration alcohol handle lacking for the methods such as crystallisation Point, according to the property and feature of Osthole, has invented the new method of 50~60% intermediate concentration alcohol processing impurity elimination crystallization, maximum Remain to limit the content and activity of Osthole.

(2) in the method that the present invention prepares Osthole from plant frutus cnidii, the refining pure of Osthole monomer component Change, invented first by adding intermediate concentration ethanol water thermosol, room temperature removal of impurities, the method for refrigerating crystallization, not only can be by Impurity is effectively dispelled, and the rate of transform of Osthole is high (about 88%).

(3) due to Osthole poorly water-soluble, it is unfavorable for directly applying to cosmetics, the mode frequently with inclusion enters it Row processing, otherwise its cosmetics prepared is also easy to produce pinch mud sense and active component difficulty is absorbed by the skin.The present invention is ingenious to be make use of Many phase compositions prepared by the characteristic and cosmetics of Osthole, Osthole is first dissolved or dissolved by heating and is permitted in preparing cosmetics Perhaps in fat-soluble excipient or carrier, its Osthole can reach without inclusion and overcome above-mentioned deficiency, greatly save Cost.

(4) method provided by the present invention is used, technological process is simple, easy to operate, and control of product quality is convenient, and having can Operability, is suitable for industrial safe and reliable preparation method.

Brief description of the drawings

Fig. 1 is the Osthole that embodiment 1 is prepared from cnidium monnieri Cnidium monnieric (L.) Cusson. fruit Photo figure.

Fig. 2 is the cnidium monnieri that embodiment 1 is prepared from cnidium monnieri Cnidium monnieric (L.) Cusson. fruit Sub- element HPLC chromatogram；Wherein, figure A is Osthole reference substance, and figure B is cnidium monnieri Cnidium monnieric (L.) Cusson. Fruit in the Osthole sample for preparing.

Fig. 3 is the Osthole photo figure that embodiment 2 is prepared from cnidium monnieri Cnidium formosanum YABE.

Fig. 4 is the Osthole HPLC chromatogram that embodiment 2 is prepared from cnidium monnieri Cnidium formosanum YABE Figure；Wherein, figure A is Osthole reference substance, the Osthole for scheming B to prepare in cnidium monnieri Cnidium formosanum YABE Sample.

Fig. 5 is the Osthole that embodiment 1 is prepared from cnidium monnieri Cnidium monnieric (L.) Cusson. fruit Photo figure.

Fig. 6 is the cnidium monnieri that embodiment 1 is prepared from cnidium monnieri Cnidium monnieric (L.) Cusson. fruit Sub- element HPLC chromatogram；Wherein, figure A is Osthole reference substance, and figure B is cnidium monnieri Cnidium monnieric (L.) Cusson. Fruit in the Osthole sample for preparing.

Embodiment

With reference to embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are not limited In this.

Embodiment 1

Extraction prepares Osthole one from plant cnidium monnieri Cnidium monnieric (L.) Cusson. fruit

(1) extraction of Osthole

Fruit using plant cnidium monnieri Cnidium monnieric (L.) Cusson. is cleaned as raw material, by drying, It is ground into the coarse powder of about 20 mesh.Frutus cnidii coarse powder 1000g is taken, the concentration for adding 10 times of amounts (10L) is that percent by volume is 85% Ethanol water, be sufficiently stirred for mixing, be heated to 80 DEG C of refluxing extractions 2 hours, filtering, filtrate is stand-by；Filter residue continuously adds 8 85% (v/v) ethanol water of (8L) times is measured, is sufficiently stirred for mixing, is heated to 80 DEG C of refluxing extractions 1.5 hours, filter, filter Liquid is stand-by；Filter residue continuously adds 85% (v/v) ethanol water of 8 times of amounts (8L), is sufficiently stirred for mixing, and is heated to 80 DEG C of backflows Extract 1.5 hours, filtering, filtrate merge with preceding secondary filtrate Osthole extract solution, it is stand-by；Filter residue is discarded.

(2) separation of Osthole runic

The extract solution of above-mentioned Osthole is taken, ethanol is recovered under reduced pressure to the greatest extent, concentrate (about 800mL) is obtained, plus 60 DEG C of heat Water is adjusted to 1500mL solution, is sufficiently stirred for mixing, is let cool to room temperature, put in about 5 DEG C of refrigerating chambers, stands 28 hours, 4000r/min centrifuges 20min, and abandoning supernatant collects precipitation, obtains the crude extract precipitation about 185g of Osthole.

(3) polishing purification of Osthole monomer component

Take the crude extract of above-mentioned Osthole to precipitate, add ethanol water 2000ml of the concentration for 55% (v/v), 60 DEG C After heating, stirring and dissolving, filtering discards precipitation, and filtrate is put to room temperature (about 25 DEG C), stands 8 hours, and filtering discards precipitation；Filter Liquid is put in 2 DEG C of refrigerating chambers, is stood 72 hours crystallizations, is refiltered separation, discards filtrate, and collection crystallizes to obtain Osthole coarse-grain 22.8g；Reprocessing recrystallization once, obtains the polishing purification crystallization 20.3g of Osthole as stated above for gained crystallization.By pharmacopeia The content assaying method of Osthole in Fructus Cnidii is determined, and the purity content for measuring Osthole is 98.9%, its Osthole Photo figure and measure chromatogram are shown in Fig. 1 and Fig. 2 respectively.Content of Osthole is 2.31%, frutus cnidii in raw material frutus cnidii (fruit) The plain rate of transform is 87.9%.

Comparative example 1

Fruit using plant cnidium monnieri Cnidium monnieric (L.) Cusson. is raw material, by patent of invention A kind of method of CN201010527673.4 " method that Osthole is extracted from Cnidium Monnieri " embodiment 3, takes cnidium monnieri Sub- coarse powder 1000g, extraction prepares Osthole, obtains the polishing purification crystallization 10.2g of Osthole, measures the purity of Osthole Content is 99.1%, and the Osthole rate of transform is 44.2%.

Embodiment 2

Extraction prepares Osthole from cnidium monnieri Cnidium formosanum YABE fruit

(1) extraction of Osthole

Fruit using plant cnidium monnieri Cnidium formosanum YABE is cleaned, by dry, pulverize as raw material Into the coarse powder of about 18 mesh.Frutus cnidii coarse powder 1000g is taken, 85% (v/v) ethanol water of 10 times of amounts (10L) is added, fully stirs Mixing is mixed, 80 DEG C of refluxing extractions are heated to 2 hours, filtering, filtrate is stand-by；Filter residue continuously adds 85% (v/v) of 8 times of amounts (8L) Ethanol water, is sufficiently stirred for mixing, is heated to 80 DEG C of refluxing extractions 1.5 hours, filtering, and filtrate is stand-by；Filter residue continuously adds 8 85% (v/v) ethanol water of (8L) times is measured, is sufficiently stirred for mixing, is heated to 80 DEG C of refluxing extractions 1.5 hours, filter, filter Liquid merge with preceding secondary filtrate Osthole extract solution, it is stand-by；Filter residue is discarded.

(2) separation of Osthole runic

The extract solution of above-mentioned Osthole is taken, ethanol is recovered under reduced pressure to the greatest extent, concentrate (about 750mL), plus 60 DEG C of hot water are obtained Adjust to 1500mL solution, be sufficiently stirred for mixing, let cool to room temperature, put in about 5 DEG C of refrigerating chambers, stand 26 hours, 4000r/ Min centrifuges 20min, and abandoning supernatant collects precipitation, obtains the crude extract precipitation about 181g of Osthole.

(3) polishing purification of Osthole monomer component

Take the crude extract of above-mentioned Osthole to precipitate, add ethanol water 2000ml of the concentration for 55% (v/v), 60 DEG C After heating, stirring and dissolving, filtering discards precipitation, and filtrate is put to room temperature (about 25 DEG C), stands 7 hours, and filtering discards precipitation；Filter Liquid is put in 1 DEG C of refrigerating chamber, is stood 72 hours crystallizations, is refiltered separation, discards filtrate, and collection crystallizes to obtain Osthole coarse-grain 21.3g；Reprocessing recrystallization once, obtains the polishing purification crystallization 19.7g of Osthole as stated above for gained crystallization.By pharmacopeia The content assaying method of Osthole in Fructus Cnidii is determined, and the purity content for measuring Osthole is 99.1%, its Osthole Photo figure and measure chromatogram are shown in Fig. 3 and Fig. 4 respectively.Content of Osthole is 2.27%, frutus cnidii in raw material frutus cnidii (fruit) The plain rate of transform is 86.8%.Its Osthole photo figure and measure chromatogram are shown in Fig. 3 and Fig. 4 respectively.

Comparative example 2

Fruit using plant cnidium monnieri Cnidium formosanum YABE is raw material, by patent of invention CN 2010 The method of the embodiment 3 of a kind of 10527673.4 " methods that Osthole is extracted from Cnidium Monnieri ", takes frutus cnidii coarse powder 1000g, extraction prepares Osthole, obtains the polishing purification crystallization 10.8g of Osthole, and the purity content for measuring Osthole is 98.8%, the Osthole rate of transform is 47.6%.

Embodiment 3

Extraction prepares Osthole two from plant cnidium monnieri Cnidium monnieric (L.) Cusson. fruit

(1) extraction of Osthole

Fruit using plant cnidium monnieri Cnidium monnieric (L.) Cusson. is cleaned as raw material, by drying, It is ground into the coarse powder of about 15 mesh.Frutus cnidii coarse powder 1000g is taken, 90% (v/v) ethanol water of 10 times of amounts (10L) is added, fills Divide and stir and evenly mix, be heated to 60 DEG C of ultrasounds (250Hz) and extract 1.5 hours, filtering, filtrate is stand-by；Filter residue continuously adds 8 times of amounts 90% (v/v) ethanol water of (8L), is sufficiently stirred for mixing, and is heated to 60 DEG C of ultrasounds (250Hz) and extracts 1.0 hours, filtering, Filtrate is stand-by；Filter residue continuously adds 90% (v/v) ethanol water of 8 times of amounts (8L), is sufficiently stirred for mixing, and is heated to 60 DEG C and surpasses Sound (250Hz) extract 1.0 hours, filtering, filtrate merge with preceding secondary filtrate Osthole extract solution, it is stand-by；Filter residue Discard.

(2) separation of Osthole runic

The extract solution of above-mentioned Osthole is taken, ethanol is recovered under reduced pressure to the greatest extent, concentrate (about 700mL), plus 60 DEG C of hot water are obtained Adjust to 1800mL solution, be sufficiently stirred for mixing, let cool to room temperature, put in about 5 DEG C of refrigerating chambers, stand 28 hours, 4000r/ Min centrifugation 20min separation, abandoning supernatant collects precipitation, obtains the crude extract precipitation about 173g of Osthole.

(3) polishing purification of Osthole monomer component

Take the crude extract of above-mentioned Osthole to precipitate, add ethanol water 2000ml of the concentration for 56% (v/v), 60 DEG C After heating, stirring and dissolving, filtering discards precipitation, and filtrate is put to room temperature (about 25 DEG C), stands 8 hours, and filtering discards precipitation；Filter Liquid is put in 1 DEG C of refrigerating chamber, is stood 72 hours crystallizations, is refiltered separation, discards filtrate, and collection crystallizes to obtain Osthole coarse-grain 22.4g；Reprocessing recrystallization once, obtains the polishing purification crystallization 20.1g of Osthole as stated above for gained crystallization.By pharmacopeia The content assaying method of Osthole in Fructus Cnidii is determined, and the purity content for measuring Osthole is 99.4%, its Osthole Photo figure and measure chromatogram are shown in Fig. 5 and Fig. 6 respectively.Content of Osthole is 2.31%, frutus cnidii in raw material frutus cnidii (fruit) The plain rate of transform is 87.0%.

Embodiment 4：The inventive method extracts the Osthole In Vitro Bacteriostasis research of separation

The fruit of cnidium monnieri subsystem Umbelliferae cnidium monnieri platymiscium cnidium monnieri (Cnidium monnieric (L.) Cusson.), Chinese medicine What frutus cnidii was often referred to is exactly that this is a kind of.Another is Cnidium formosanum YABE.Two kinds of frutus cnidiis are in China various regions There is growth, but it is in the majority with former.Jiangsu, the province of Anhui two are the main products of frutus cnidii.Frutus cnidii is more as outer since ancient times Medication is mainly outer to be used to treat skin disease such as using recording seldom about its purposes in the medical works of Ancient Times in China《The legendary god of farming Book on Chinese herbal medicine warp》In have " curing mainly wet tinea, malignant sore "；《Collect letter side》There is the record of " married woman's pruritus vulvae is permitted end with frutus cnidii and lard applies it "； 《Compendium of Materia Medica》The effect such as cloudy sweat, wet tinea is recorded, the effect such as Trichomonas vaginalis, skin eczema is cured mainly.Frutus cnidii have preferably Antibacterial and anti-inflammation functions, staphylococcus aureus, staphylococcus aureus resistance bacterium, Escherichia coli, dysentery will Hayes can be suppressed The bacteriums such as bacterium, salmonella typhimurium, Pseudomonas aeruginosa, has a broad antifungal spectrum, experimental method and result are as follows:

1 materials and methods

1.1 instrument

The electro-heating standing-temperature cultivators of DY6000 II (Tianjin Stettlen Instrument Ltd.)；The double super-clean benches of HT-20-5MT (Soviet Union The safe and sound air technique Co., Ltd in state)；Micropipettor (Shanghai thermoelectricity Instrument Ltd.)；LDZX-50K autoclaves (Shanghai Sanshen Medical Instrument Co., Ltd.).

1.2 reagent

Osthole is by the inventive method by embodiment 1 from plant cnidium monnieri Cnidium monnieric (L.) Cus-son. Fruit in extract and be prepared, HPLC controls through Osthole reference substance, and containing by pharmacopeia Osthole in Fructus Cnidii Quantity measuring method is determined, and the purity content for measuring Osthole is 98.9%, its Osthole photo figure and measure chromatogram point Fig. 1 and Fig. 2 are not seen.

1.3 strain

Staphylococcus aureus (CMCC26003), staphylococcus aureus resistance bacterium (S4050), Escherichia coli (JM110), shigella dysenteriae (CMCC51252), salmonella typhimurium (CMCC50115), Pseudomonas aeruginosa (CMCC10104) Bought from Guangdong Province's Culture Collection.

2 methods and result

The preparation of 2.1 Osthole fat emulsion solutions and blank fat emulsion solution

2.1.1 the preparation of Osthole fat emulsion solution

Osthole 1.5g prepared by embodiment 1 adds 15ml soybean oil, and Osthole is heated in 60 DEG C of water-baths Hold solution completely, under conditions of logical nitrogen, soybean lecithin 1.5g is added in soybean oil, smashs complete to soybean lecithin to pieces at a high speed Dissolving, 60 DEG C of keeping temperature obtains oil phase；Take purified water 50mL to put in container, glycerine 15g is slowly added under stirring, heating rises Temperature is to 60 DEG C, and stir obtained aqueous phase；Under conditions of 60 DEG C of constant temperature and high speed dispersion and emulsion 12000rpm, by oil phase slowly Add in aqueous phase, be extremely well mixed, plus purified water is adjusted to 100mL, is put into homogenizer, regulation homogenization pressure 60Mpa, homogeneous Temperature 60 C, homogeneous 10 times, the filtering of 350 mesh filters, obtain 15mgmL^-1Osthole fat emulsion solution, it is standby.

2.1.2 the preparation of blank fat emulsion solution

15ml soybean oil is taken, under conditions of logical nitrogen, soybean lecithin 1.5g is added in soybean oil, at a high speed 12000rpm is smashed to pieces to soybean lecithin and is completely dissolved, and 60 DEG C of keeping temperature obtains oil phase；Take purified water 50mL to put in container, stir shape Glycerine 15g is slowly added under state, 60 DEG C are heated to, stir obtained aqueous phase；In 60 DEG C of constant temperature and high speed dispersion and emulsion Under conditions of 12000rpm, oil phase is slowly added into aqueous phase, is extremely well mixed, plus purified water is adjusted to 100mL, is put into homogeneous In machine, regulation homogenization pressure 60Mpa, 60 DEG C of homogenizing temperature, homogeneous 10 times, 350 mesh filters filtering obtain blank fat emulsion solution, It is standby.

The preparation of 2.2 bacterium solutions

The single bacterium colony of different strain is inoculated in LB nutrient solutions respectively, 37 DEG C of 18~24h of constant-temperature table Zengjing Granule. A small amount of bacterium solution is further taken out, 10 are diluted to 0.9% sodium chloride solution of sterilizing⁷~10⁸CFU·mL^-1

2.3 bacteriostatic experiment

2.3.1 quick paper disk method

Make the drug sensitive test paper containing various concentrations Osthole and a diameter of 5mm.The different μ L of strain 100 are coated on On LB culture mediums, after media surface is dried, the scraps of paper are affixed on after media surface, 37 DEG C of culture 24h, observes and determines suppression Collarium diameter, is repeated 3 times, the negative control group blank fat emulsion solution scraps of paper.

2.3.2 doubling dilution

Culture medium 2.0mL needed for being separately added into each strain in 5mL band plug sterilizing test tubes, every kind of each 8 pipe of strain.By 1~8 Numbering, adds Osthole fat emulsion solution 2mL in the 1st pipe respectively, and 2.0mL is taken out after mixing and is put into the 2nd pipe, using two times Dilution method, it is respectively 7.50 that decoction is absorbed into 1: 2,1: 4,1: 8,1: 16,1: 32,1: 64 serial solution, i.e. content of dispersion successively, 3.75,1.88,0.94,0.47,0.24mgmL^{- 1}.It is about 5 × 105CFUmL that each pipe, which adds concentration,^{- 1}Each cause of disease indicate Bacterium bacterium solution 0.2mL, while setting negative control pipe (being free of bacterium solution containing decoction) respectively, positive control pipe (contains bacterium solution not drug containing). Cultivated in 37 DEG C of incubators and growth situation is observed after 24h.Pathogen density calculating method:Bacterium colony on observation flat board is formed Unit (CFU).The concentration of CFU≤5 is Mlc, marks "-" number, the concentration of CFU >=5 is without Mlc, mark "+", the Cmin for producing bacteriostasis represents that the pipe dose is the minimum inhibitory concentration (MIC) of by reagent.

2.4 results are with discussing

2.4.1 result

Paper disk method determines Osthole and is shown in Table 1 to the bacteriostatic activity of 6 kinds of bacterium；Doubling dilution determines Osthole to 6 kinds The MIC value of bacterium is shown in Table 2.

Average inhibition zone diameter mm of the Osthole of table 1 to 6 kinds of bacterium

MIC value n=3, mgmL of the Osthole of table 2 to 6 kinds of bacterium^-1

Understood by table 1,2, Osthole has obvious bacteriostasis to 6 kinds of bacterium, and makees as the increase of concentration is antibacterial With enhancing.Wherein to the bacteriostasis of staphylococcus aureus, staphylococcus aureus resistance bacterium, Pseudomonas aeruginosa and Escherichia coli Most strong, MIC is 0.94mgmL^-1；To salmonella typhimurium bacteriostasis secondly, MIC is 1.88mgmL^-1；To dysentery Shigella bacteriostasis is most weak, and MIC is 3.75mgmL^-1。

2.4.2 discuss

The present invention has investigated Osthole to staphylococcus aureus, golden yellow with quick paper disk method and test tube doubling dilution Staphylococcus drug-fast bacteria, Escherichia coli, shigella dysenteriae, salmonella typhimurium, Pseudomonas aeruginosa bacteriostasis.Result is ground to show Show Osthole to staphylococcus aureus, staphylococcus aureus resistance bacterium, Pseudomonas aeruginosa and Escherichia coli bacteriostasis most By force, next to that salmonella typhimurium, acts on weaker to shigella dysenteriae.As a result point out：Osthole and its cosmetics system Agent has stronger antibacterial and anti-inflammation functions in clinic, can suppress staphylococcus aureus, staphylococcus aureus resistance bacterium, large intestine bar The bacteriums such as bacterium, shigella dysenteriae, salmonella typhimurium, Pseudomonas aeruginosa；There are significant anti-inflammation, function of removing acne.

Embodiment 5：The inventive method extracts the Osthole anti-inflammatory analgetic effect experiment of separation

Osthole fat emulsion solution has preferable antibacterial action, can suppress staphylococcus aureus, staphylococcus aureus The bacteriums such as drug-fast bacteria, Escherichia coli, shigella dysenteriae, salmonella typhimurium, Pseudomonas aeruginosa；In addition, Osthole has Anti-inflammatory analgetic is acted on, and experimental method and result are as follows:

1 reagent

The preparation of 1.1 Osthole fat emulsion solutions and blank fat emulsion solution:Be performed as described above in example 4 it is 2.1 made, for abdomen Chamber injection.

1.2 indocin:Shanghai pharmaceuticals product, before use, is made 5mg/ml, for intraperitoneal injection.

2 experimental animals and environment

Kunming mice, male, 20 ± 2g of body weight, Guangdong Medical Lab Animal Center provides, animal quality verification of conformity Numbering：No.44411600000901.Experimental animal uses credit number：SYXK (Guangdong) 2013-0002.

Experimental situation condition:23 DEG C ± 2 DEG C of temperature, relative humidity 60% ± 10%, interior is led to based on natural lighting Wind is good, and environment is kept quite, SPF grades of experimental animal environmental facilities.

120 mouse are divided into 2 big group, each big group 60；Each big group is divided into the control of blank fat emulsion solution again Group, indocin make positive controls, Osthole fat emulsion solution it is small, in, big three dosage administration groups, each group 12.

3 experimental methods

With reference to bureau of drug administration of Ministry of Health of the People's Republic of China chief editor《New drug preclinical study guideline collects》Method Two

3.1 mouse writhings are tested

After mice group, negative control is made with blank fat emulsion solution, indocin makees positive control, intraperitoneal injection, After 40~60 minutes, 0.6% acetic acid 0.2m1 is injected intraperitoneally, mouse writhing number of times is observed in 15 minutes behind.Show between comparative group Write sex differernce (explanation:Mouse writhing reaction refers to the belly indent that mouse produces because being pierced emblem by pain, and trunk is stretched with back leg , the phenomenon of hips up).

3.2 mouse auricular conchas cause scorching method

After mice group, negative control is made with blank fat emulsion solution, indocin makees positive control, drug injection abdominal cavity Injection, after being administered 30 minutes, dimethylbenzene 0.03-0.05ml is put on clearly inside and outside mouse right ear auricular concha.Mouse is put to death after 3 hours and is taken Bottom right auricular concha, while remove the control of left auricular concha, 8mm card punch is respectively after left and right ear lays round auricle, on assay balance Weigh, calculate swelling.(swelling=auris dextra auricular concha weight-left ear auricular concha weight).Significant difference between comparative group.

4 experimental results

The Osthole Dichlorodiphenyl Acetate of table 3 induce small people's writhing inhibitory action (N=12)

^*P<0.05,^**P<0.O1 is compared with blank fat emulsion solution group.

The Osthole paraxylene of table 4 induction mouse auricular concha swelling inhibitory action (N=12)

^*P<0.05,^**P<0.01 compared with blank fat emulsion solution group.

Test result indicates that, the mouse writhing of Osthole Dichlorodiphenyl Acetate induction has significant inhibitory action, and action intensity It is related to dosage.And big or middle 2 dosage groups are better than indocin.It the results are shown in Table 3.

Experimental result shows again：Osthole has significant inhibitory action to mouse auricular concha swelling, and inhibitory action is with dosage Increase and increase and big or middle 2 dosage action intensities are better than indocin group.It the results are shown in Table 4.5 conclusions

This test result indicates that, there is Osthole of the invention significant anti-inflammatory analgetic to act on.Effect is better than indocin. It can be used for preparing natural cosmetics or antiinflammation pain-stopping pharmaceutical.

Embodiment 6：The inventive method extracts the Osthole anti-allergy action experiment of separation

Osthole can be used for another important function of cosmetics to be its antiallergic action, herein using mouse and rat to grind Study carefully object, reacted by suppressing mouse models of passive skin irritability (PCA), suppress rat peritoneal mast cells degranulation, it was demonstrated that this hair Bright Osthole has significant anti-allergic effects, is now summarized as follows result：

1 reagent

The preparation of 1.1 Osthole fat emulsion solutions and blank fat emulsion solution:Be performed as described above in example 4 it is 2.1 made, for mouth Clothes and intraperitoneal injection are used.

1.2 root of Chinese trichosanthes parenteral solutions：Shanghai biochemical-pharmaceutical factory product, is made into 0.5% Evans blue, physiological saline 1.00mg/ml, 1.25mg/ml, injection；Gel aluminum hydroxide is bought for market.

1.3 antiserum：Edited by Xu Shuyun etc.《Pharmacological experimental methodology》(the 1st edition, Beijing:People's Health Publisher, 1982:923) method is produced.

1.4 nasmil parenteral solutions：Shanghai biochemical-pharmaceutical factory product, 5.8mg/ml, injection are made into physiological saline.

2 experimental animals

Kunming mice (20 ± 2g of body weight), Wister rats (body weight 200g ± 20g), Guangdong Medical Lab Animal Center There is provided, animal quality verification of conformity numbering：No.44411600000901.Experimental animal uses credit number：SYXK (Guangdong) 2013-0002。

3 experimental methods

3.1 mouse models of passive skin irritability (PCA) are tested

Healthy mice is taken, belly is lost hair or feathers quiet after tail after intracutaneous injection antiserum 0.03ml (10 times of serum-dilution), 48h Arteries and veins injects root of Chinese trichosanthes parenteral solution (1.25mg/ml) 0.2mL, carries out sacrificed by decapitation animal after attack 30min, overturns skin of abdomen, Measurement locus coeruleus diameter simultaneously cuts color spot skin graft, and 5ml acetone physiological saline (7 is dipped in respectively:3), eluted after 24h, use spectrophotometric Meter reads optical density in 610nm wavelength colorimetrics.

3.2 rat peritoneal mast cells degranulations are tested

Healthy rat is taken, sole injection root of Chinese trichosanthes-gel aluminum hydroxide suspension, 4 sole co-injection 0.4ml are that is, every Mouse injects root of Chinese trichosanthes 1mg, 15d after injection, is injected intraperitoneally and is tested etherization after medicine, 0.5h, femoral artery sacrificed by exsanguination animal, Abdominal cut skin, to intraperitoneal injection pH 6.9 HankShi liquid (without phenol red) 10ml, includes heparin 0.005%, gently Abdomen massage 2min, then opens abdominal cavity, and peritoneal fluid is collected in test tube with dropper, puts after ice bath cooling with 1000r/min 15min is centrifuged, supernatant is abandoned, sediment is suspended from 2ml HankShi liquid, refrigerator is put standby.

Above-mentioned mast cell suspension 0.5ml, plus man equivalent antigen liquid (containing root of Chinese trichosanthes 1mg/ml) are taken, 37 DEG C are incubated 15min, 0.125% neutral red staining, 100 mast cells of high power Microscopic observation calculate medicine to fertilizer by the formula of (1-a/q) × 100% The suppression percentage of maxicell degranulation.A and q represent administration group and control group degranulation cell number respectively in formula.

4 experimental results

The inhibitory action that 4.1 Ostholes are reacted mouse PCA

Three groups are divided into after taking mouse 30, belly intracutaneous injection antiserum, control group gavages blank fat emulsion solution, separately Two groups once gavage Osthole 100mg/kg and 200mg/kg respectively fat emulsion solution.The locus coeruleus diameter and light of three groups of animals Density is shown in Table 5.As a result show that Osthole there are mouse PCA reactions to act on compared with high inhibition.

The Osthole of table 5 to mouse models of passive skin irritability (PCA) inhibiting rate influence (N=10)

Osthole mg/kg	Locus coeruleus diameter (mm)	Optical density
			0	1.21±0.23	0.167±0.07
100	0.51±0.25*	0.074±0.04*
			200	0.43±0.28**	0.051±0.02**

^*P<0.05,^**P<0.01 compared with blank fat emulsion solution group.

Inhibitory action of 4.2 Ostholes to mast cell degranulation

Rat 40 is taken, four groups are divided into, control group distinguishes intraperitoneal injection of saline 0.7ml/ and nasmil Only, Osthole fat emulsion solution 0.7mL/ and 1.4mL/ is injected intraperitoneally only in administration group to 0.7ml/ respectively.To root of Chinese trichosanthes antigen Caused rat peritoneal mast cells degranulation, the inhibiting rate of nasmil is 64.0%, and the inhibiting rate of Osthole is distinguished For 57.7% and 79.1%, compared that there were significant differences with control group, show de- of Osthole in Rats peritoneal cavity mast cells Grain has more significant inhibitory action.Its experiment the results are shown in Table 6.

The Osthole of table 6 to the inhibitory action of mast cell degranulation (N=10)

5 discuss

Experiment is proved：PCA reaction of the Osthole to mouse 48h has stronger inhibitory action, shows that it can substantially suppress The formation of IgE antibody, has inhibitory action to the mast cell degranulation in allergy.Osthole is anti-for preparing resistance state The cosmetics answered play an important roll.

Embodiment 7：Osthole cosmetics preparating example

(1) the preparation of face cream

Prescription：

Preparation technology:

Take Osthole, cetearyl glucoside, hexadeca-octadecyl alcohol, caprylic/capric glyceryl ester, jojoba oil, palm fibre Palmitic acid isopropyl propionate, glycerine are mixed and heated to 60 DEG C, as A phases, are incubated standby；Extracting liquorice acid dipotassium, addition xanthan gum, card ripple 940th, Sodium Hyaluronate, deionized water, are mixed and heated to 60 DEG C, are uniformly dispersed, as B phases, are incubated standby；By A phases in homogeneous Under be slowly added to be uniformly dispersed and be heated in 80 DEG C of B phases in advance, add ring dimethicone 345 homogeneous 3 minutes, add Sodium hydroxide adjusts pH6-7, and stirring is cooled to 50 DEG C, adds essence, stirs, is cooled to 35 DEG C and produces.

(2) the preparation of emulsion

Prescription：

Preparation technology:

Osthole, cyclopentasiloxane 995, vaseline, Tween-80, Vitwas E, glycerine is taken to be mixed and heated to 60 DEG C, as A phases, it is incubated standby；Extracting liquorice acid dipotassium, add xanthan gum, Carbopol Ultrez 10,1,3-BDO, Deionized water, is mixed and heated to 60 DEG C, is uniformly dispersed, as B phases, is incubated standby；A phases are slowly added in advance under homogeneous It is uniformly dispersed and is heated in 80 DEG C of B phases, homogeneous 3 minutes adds triethanolamine regulation pH6-7, and stirring is cooled to 50 DEG C, plus Enter essence, preservative, stir, be cooled to 35 DEG C and produce.

(3) the preparation of eye cream

Prescription：

Preparation technology:

Extracting liquorice acid dipotassium, adds 1,3-BDO, green-tea extract, disodium ethylene diamine tetraacetate, deionized water, disperses Uniformly, 50 DEG C are heated to, stirring is until limpid；Elastomer silicone is added slowly with stirring, it is standby as A phases；Take cnidium monnieri Sub- element, DC 245, jojoba oil, Vitwas E, Retinol Palmitate are mixed and heated to 60 DEG C, are used as B phases, insulation It is standby；A phases are slowly added under homogeneous to be uniformly dispersed and be heated in 60 DEG C of B phases in advance, homogeneous 3 minutes is cooled to 35 DEG C produce.

(4) the preparation of gel

Prescription：

Preparation technology:

Take Osthole, dipotassium glycyrrhizinate, add polyacrylic acid, glycerine, 1,3-BDO, fucoidan, thoroughly The sour sodium of bright matter, dispersed with stirring is uniform, standby as A phases；Remove ionized water to add A phases, be uniformly dispersed, be heated to 60 DEG C, addition Triethanolamine adjusts pH6-7, and stirring is cooled to 50 DEG C and adds essence, methyl-isothiazol beautiful jade ketone, stirs, is cooled to 35 DEG C and produces.

Above-described embodiment is preferably embodiment, but embodiments of the present invention are not by above-described embodiment of the invention Limitation, other any Spirit Essences without departing from the present invention and the change made under principle, modification, replacement, combine, simplification, Equivalent substitute mode is should be, is included within protection scope of the present invention.

Claims (10)

1. a kind of method that Osthole is prepared from plant, it is characterised in that comprise the following steps：

(1) extraction of Osthole：Using the fruit of samphire as raw material, cleaned, dried, crushed, obtain coarse powder；With The ethanol solution heating extraction or heating temperature leaching ultrasonic extraction of percent by volume 65%~95%；After having extracted, separation of solid and liquid is obtained To the extract solution containing Osthole；

(2) separation of Osthole runic：The extract solution containing Osthole is taken, ethanol is reclaimed to complete, obtains concentrate；Dense Temperature is added in contracting liquid to be dissolved for 40~80 DEG C of hot water, then cools down, more than 24 hours are stood then at 1~25 DEG C, Gu Liquid is separated, and discards liquid, is collected solid, is obtained the crude extract containing Osthole；

(3) polishing purification of Osthole monomer component：By step 1. or 1. and 2. step carries out polishing purification；

1. crystallize for the first time：The ethanol water that concentration is percent by volume 50~60% is added in Osthole crude extract, Dissolve by heating, be cooled to room temperature, stand for the first time, separation of solid and liquid takes liquid, discards solid；Liquid obtained by separation of solid and liquid in 0~ Second of standing is fully separated out to crystal in 10 DEG C of environment, then separation of solid and liquid, discards liquid, collects crystallization；

2. recrystallize；1. obtained crystallization is recrystallized according still further to step, the polishing purification crystallization of Osthole is obtained.

2. the method for Osthole is prepared from plant according to claim 1, it is characterised in that：

Ethanol solution described in step (1) is the ethanol solution that concentration is percent by volume 85~90%；

Hot water described in step (2) is the hot water that temperature is 50~70 DEG C；

Step (3) 1. described in ethanol water concentration be percent by volume 53~57%.

3. the method for Osthole is prepared from plant according to claim 1, it is characterised in that：

The number of times of heating extraction described in step (1) is 2~3 times；Comprise the following steps that：When extraction time is 2 times：For the first time The ethanol solution equivalent to 10 times of volumes of meal quality is added, is sufficiently stirred for mixing, it is small in 70~100 DEG C of refluxing extractions 1~3 When；Second added the ethanol solution equivalent to 8 times of volumes of meal quality, in 70~100 DEG C of refluxing extractions 0.5~2 hour； When extraction time is 3 times：It is preceding twice with extraction time be 2 times when the step of, third time add equivalent to 8 times of volumes of meal quality The ethanol solution of amount, in 70~100 DEG C of refluxing extractions 0.5~2 hour；After having extracted, separation of solid and liquid merges 2~3 times and separated The liquid arrived, obtains the extract solution containing Osthole；

The number of times of heating temperature leaching ultrasonic extraction described in step (1) is 2~3 times；Comprise the following steps that：Extraction time is 2 times When：The ethanol solution equivalent to 10 times of volumes of meal quality is added for the first time, is sufficiently stirred for mixing, it is super in 60~80 DEG C of temperature leachings Sound is extracted 0.5~2 hour；Second of addition surpasses equivalent to the ethanol solution of 8 times of volumes of meal quality in 60~80 DEG C of temperature leachings Sound is extracted 0.5~1.5 hour；When extraction time is 3 times：It is preceding twice with extraction time be 2 times when the step of, third time add phase When in the ethanol solution of 8 times of volumes of meal quality, ultrasonic extraction is soaked 0.5~1.5 hour in 60~80 DEG C of temperature；After having extracted, Separation of solid and liquid, merges 2~3 isolated liquid, obtains the extract solution containing Osthole.

4. the method for Osthole is prepared from plant according to claim 3, it is characterised in that：

The ultrasound condition in heating temperature leaching ultrasonic extraction described in step (1) is 200~300Hz.

5. the method for Osthole is prepared from plant according to claim 1, it is characterised in that：

The condition of standing described in step (2) is to stand 24~30h in 2~8 DEG C；

Step (3) 1. described in first time stand time be 7~8 hours；

Step (3) 1. described in second standing time be more than 48 hours；

Step (3) 1. described in second standing temperature be 0~8 DEG C.

6. the method for Osthole is prepared from plant according to claim 1, it is characterised in that：

The addition of hot water described in step (2) by extracting of obtaining of the 1kg raw materials volume that concentrate added after hot water for 1~ 2L is calculated；

Step (3) 1. described in the crude extract that obtains by 1kg raw materials of addition of ethanol water to match 1~3L ethanol water-soluble Liquid is calculated.

7. the method for Osthole is prepared from plant according to claim 1, it is characterised in that：

Samphire described in step (1) is cnidium monnieri Cnidium monnieric (L.) Cusson. or Cnidium formosanum YABE；

Coarse powder described in step (1) is the coarse powder of 15~20 mesh；

Step (3) 2. described in recrystallization number of times be 1 time.

8. application of the method described in any one of claim 1~7 in cosmetics are prepared.

9. a kind of refined Osthole, it is characterised in that：Prepared by the method described in any one of claim 1~7, Purity content >=98% (w/w).

10. application of the refined Osthole in cosmetics are prepared described in claim 9.