CN108159195B - Acer truncatum extract, preparation method and application thereof - Google Patents
Acer truncatum extract, preparation method and application thereof Download PDFInfo
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- 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)
- A61K36/85—Verbenaceae (Verbena family)
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- 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
- A61K2236/35—Extraction with lipophilic solvents, e.g. Hexane or petrol ether
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- 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
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- 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
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- 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
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
Abstract
The invention discloses a Acer truncatum extract, a preparation method and application thereof, and belongs to the technical field of natural product extraction. The Acer acutangum extract contains mixture of carnosol, rosmanol and isorosmanol. The invention also discloses a preparation method and application of the Acer truncatum extract. The Acer acutangum extract contains a mixture of carnosol, rosmanol and isorosmanol, and the components are diterpene phenol components which have better activities of oxidation resistance, cancer resistance, inflammation resistance and the like. The pharmacodynamic study on the Acer truncatum extract provides a wider space for further application of the Acer truncatum extract.
Description
Technical Field
The invention relates to a Acer truncatum extract, a preparation method and application thereof, and belongs to the technical field of natural product extraction.
Background
Nitric Oxide (NO) is a free radical that functions as both a second messenger and a neurotransmitter and as an effector molecule that mediates the pathological and physiological processes of a variety of diseases such as inflammation and immunity. NO is an important inflammatory factor in acute and chronic inflammation. A plurality of research results show that the NO level in the synovial fluid and serum of the patients with rheumatism is obviously higher than that of normal people. NO is not only directly involved in rheumatism as an inflammation medium, but also can promote the release of other inflammatory factors, further promote the destruction of articular cartilage and aggravate the disease. If the production of NO is reduced, protection against joint inflammation may be provided.
Acer acutus (Callicarpa longissima) is a plant of Callicarpa of Verbenaceae, and is named as Zhanfeng, Chuangu maple, xue Tu, Niangguang, etc. Distributed in Jiangxi, Fujian, Taiwan, Guangdong, Guangxi, Sichuan and so on. It is pungent, slightly bitter and warm in nature, and has effects of stopping bleeding, relieving pain, removing blood stasis, relieving swelling, and dispelling pathogenic wind, and can be used for treating traumatic hemorrhage, hemoptysis, puerperal rheumatalgia, quadriplegia, and rheumatalgia.
However, at present, the research on the acer truncatum is less at home and abroad, and the application of the active components of the acer truncatum is more rarely reported. Therefore, research on extraction and purification processes of the active components of acer truncatum and related applications thereof is needed to overcome the above defects.
Disclosure of Invention
The invention aims to provide a Acer truncatum extract. The Acer acutangum extract contains a mixture of carnosol, rosmanol and isorosmanol, and the components are diterpene phenol components which have various activities such as better oxidation resistance, cancer resistance, anti-inflammation and the like. The pharmacodynamic study on the Acer truncatum extract provides a wider space for further application of the Acer truncatum extract.
The technical scheme for solving the technical problems is as follows: the Acer acutangum extract contains a mixture of carnosol, rosmanol and isorosmanol, wherein the weight percentage of the carnosol in the Acer acutangum extract is 53.0-82.6%, the weight percentage of the rosmanol in the Acer acutangum extract is 4.5-12.4%, and the weight percentage of the isorosmanol in the Acer acutangum extract is 2.1-8.0%
Carnosol, rosmanol and isorosmanol are diterpene phenol components. Pharmacological research shows that diterpene phenol components in natural products have better drug effect activities of antioxidation, antibiosis, anticancer, neuroprotection, anti-inflammation and the like, wherein in the aspect of anti-inflammatory activity, the components can inhibit the formation of lipopolysaccharide, target on microsomal prostaglandin E2 synthase and inhibit the synthesis of prostaglandin E2 synthase; also can exert anti-inflammatory activity by inhibiting Src/NF-kB signal channels and the expression and release of inflammatory factors IL-1 beta and IL-6.
The inventor of the application finds that the acer truncatum extract is rich in diterpene phenolic components such as carnosol, rosmanol, isorosmanol and the like, and pharmacological results show that the extract has good anti-inflammatory activity. This discovery of the present application will provide a broader space for further applications of acer truncatum extracts.
The second object of the present invention is to provide a method for preparing the acer truncatum extract. The invention adopts the principle of similar intermiscibility, utilizes an organic solvent with low and medium polarity to quickly extract diterpene phenol components from acer truncatum, and effectively removes pigments and enriches the diterpene phenol components by a column chromatography separation technology.
The technical scheme for solving the technical problems is as follows: a preparation method of Acer truncatum Bunge extract comprises the following steps:
step 1: preparation of Acer acutangula crude extract
Taking dried Acer acutus powder, adding organic solvent with the mass 10-20 times of that of the Acer acutus powder, reflux-extracting for 3 times, filtering, combining filtrates, and concentrating the filtrate under reduced pressure to obtain extract, i.e. Acer acutus crude extract;
step 2: purification of
Purifying the Acer truncatum crude extract obtained in the step 1 by silica gel column chromatography, removing oil and fat components by using petroleum ether, eluting by using a mixed solvent of the petroleum ether and ethyl acetate with a volume ratio of (5-3) to 1 or a mixed solvent of the petroleum ether and acetone with a volume ratio of (6-4) to 1, collecting eluent, recovering the solvent, and drying to obtain a purified Acer truncatum extract;
or purifying the Acer truncatum crude extract obtained in the step 1 by reverse phase material column chromatography, removing impurities by using alcohols with the volume concentration of 40-50%, eluting by using alcohols with the volume concentration of 75-95%, collecting eluent, recovering the solution, and drying to obtain the purified Acer truncatum extract.
On the basis of the technical scheme, the invention can be further improved as follows.
Further, the Acer acutangum powder in the step 1 is obtained by drying stems, branches and leaves of the Acer acutangum serving as raw materials and then crushing the dried material to 2 mm.
The adoption of the further beneficial effects is as follows: the proper particle size is not only beneficial to the extraction of effective components, but also beneficial to the filtration of the extracting solution.
Further, the organic solvent in step 1 is one of dichloromethane, chloroform, ethyl acetate and acetone.
The adoption of the further beneficial effects is as follows: the organic solvent is a medium-low polarity solvent, and can be used as an extraction solvent to effectively and rapidly extract diterpene phenol components.
Further, the temperature of the extraction in the step 1 is 50-70 ℃ and each time is 1 hour.
The adoption of the further beneficial effects is as follows: the extraction temperature is mild, and the active ingredients are prevented from changing due to high temperature.
Further, the pore size of the filtration in the step 1 is 30-70 μm.
The adoption of the further beneficial effects is as follows: can effectively and quickly filter the extracting solution.
Further, the specific gravity of the extract in the step 1 at 65 ℃ is 1.02-1.08.
Further, the medium of the reversed phase column chromatography in the step 2 is one of C18, C8, C4 or MCI.
Wherein the particle size of the C18 is 40-60 μm, the particle size of the C8 is 40-75 μm, the particle size of the C4 is 40-60 μm, and the particle size of the MCI is 75-150 μm.
The adoption of the further beneficial effects is as follows: can effectively remove impurities such as pigment in the acer truncatum raw material, and the product has good color and luster.
Further, the alcohols in step 2 are all methanol or ethanol.
The adoption of the further beneficial effects is as follows: the methanol or ethanol solution with proper concentration can effectively resolve the diterpene phenol components in the reversed phase material, so that the diterpene phenol components are more concentrated.
The third purpose of the invention is to provide the application of the acer truncatum bunge extract. The Acer acutangum extract can be applied to preparing anti-inflammatory drugs or anti-inflammatory auxiliary drugs, and has wide application space.
The technical scheme for solving the technical problems is as follows: an application of the Acer acutangum extract in preparing anti-inflammatory drugs or anti-inflammatory auxiliary drugs.
The mouse ear swelling model induced by xylene causes the dissolution and release of certain inflammatory mediators such as histamine, kinin and fibrin, causes local vasodilatation, increases capillary permeability, infiltrates inflammatory cells, and causes ear acute exudative inflammatory edema. The model is widely used in domestic evaluation of the anti-inflammatory activity of plant extracts or synthetic compounds.
The inventor of the application adopts an inflammatory factor NO generation inhibition cell model and a xylene-induced mouse ear swelling acute inflammation model as an anti-inflammatory efficacy evaluation method. The extract is subjected to anti-inflammatory pharmacodynamic evaluation, and the result shows that the Acer truncatum extract disclosed by the invention shows good anti-inflammatory activity and can be applied to preparation of anti-inflammatory drugs or anti-inflammatory auxiliary drugs.
The invention has the beneficial effects that:
(1) the Acer acutangum extract contains a mixture of carnosol, rosmanol and isorosmanol, all the components are diterpene phenol components, and the components have various activities such as better antioxidation, anticancer, anti-inflammatory and the like. The pharmacodynamic study on the Acer truncatum extract provides a wider space for further application of the Acer truncatum extract.
(2) The invention adopts the principle of similar intermiscibility, utilizes an organic solvent with low and medium polarity to quickly extract diterpene phenol components from acer truncatum, and effectively removes pigments and enriches the diterpene phenol components by a column chromatography separation technology.
(3) The preparation method of the acer truncatum bunge extract is simple, easy to operate, wide in market prospect and suitable for large-scale production.
(4) The Acer acutangum extract can be applied to preparing anti-inflammatory drugs or anti-inflammatory auxiliary drugs, and has wide application space.
Drawings
FIG. 1 is a HPLC chromatogram of three control samples of carnosol, rosmanol and isorosmanol in example 1 of the present invention.
FIG. 2 is a HPLC chromatogram obtained in example 2 of the present invention.
FIG. 3 is a HPLC chromatogram obtained in example 3 of the present invention.
FIG. 4 is a HPLC chromatogram obtained in example 4 of the present invention.
FIG. 5 is a HPLC chromatogram obtained in example 5 of the present invention.
FIG. 6 is a HPLC chromatogram obtained in example 6 of the present invention.
Detailed Description
The principles and features of this invention are described below in conjunction with the following detailed drawings, which are given by way of illustration only and are not intended to limit the scope of the invention.
Example 1
Weighing carnosol 2.0mg, rosmanol 2.2mg and isorosmanol 2.5mg respectively, placing in a 10mL volumetric flask together as a reference, adding methanol for dissolving, and fixing the volume to scale to obtain a mixed reference to-be-detected liquid, sucking 10 μ L of the mixed reference to-be-detected liquid, and injecting into a high performance liquid phase for determination.
Liquid chromatography conditions: agilent 1200 liquid chromatograph, column: ZORBAX SB-C18(5 μm,4.6X250mm), column temperature: 30 ℃, flow rate: 1mL/min, detection wavelength: 210nm, mobile phase acetonitrile (a)/water (B): gradient elution, 0-25min, 40% -74% (A).
The results are shown in FIG. 1.
As can be seen from FIG. 1, carnosol, rosmanol and isorosmanol have good separation under the above chromatographic conditions.
Example 2
The acer truncatum extract of the embodiment contains a mixture of carnosol, rosmanol and isorosmanol, wherein the mass content of the carnosol in the acer truncatum extract is 79.6%, the mass content of the rosmanol in the acer truncatum extract is 7.8%, and the mass content of the isorosmanol in the acer truncatum extract is 6.6%.
The preparation method of the Acer truncatum extract comprises the following steps:
step 1: preparation of Acer acutangula crude extract
Drying stems, branches and leaves of the raw material Acer acutangum, and crushing to 2mm to obtain dried Acer acutangum powder.
Taking 600g of dried Acer acutus powder, adding dichloromethane which is 10 times of the mass of the Acer acutus powder, carrying out reflux extraction for 3 times at 50 ℃, each time for 1 hour, sieving by a 30-micron sieve, combining filtrates, and concentrating the filtrate under reduced pressure to an extract with the specific gravity of 1.02 at 65 ℃ to obtain 32.2g of Acer acutus crude extract.
Step 2: purification of
And (2) purifying 30g of the Acer truncatum crude extract obtained in the step (1) by silica gel column chromatography, removing oil and fat components by using petroleum ether, eluting by using a mixed solvent of the petroleum ether and ethyl acetate in a volume ratio of 5:1, collecting eluent, recovering the solvent, and drying to obtain 10.4g of the purified Acer truncatum extract.
HPLC sample test solution preparation for this example: weighing 3.6mg of the prepared Acer acutangum extract, placing into a 10mL volumetric flask, adding methanol for dissolving, and fixing the volume to scale to obtain HPLC solution to be detected, sucking 10 mu L of the solution to be detected, and injecting into high performance liquid for detection.
Liquid chromatography conditions: the same as in example 1.
The results are shown in FIG. 2.
As can be seen from fig. 2, the content of carnosol by mass% was 79.6% by HPLC analysis.
The Acer acutus extract is applied to the preparation of anti-inflammatory drugs or anti-inflammatory auxiliary drugs.
Example 3
The acer truncatum extract of the embodiment contains a mixture of carnosol, rosmanol and isorosmanol, wherein the mass content of the carnosol in the acer truncatum extract is 59.6%, the mass content of the rosmanol in the acer truncatum extract is 12.4%, and the mass content of the isorosmanol in the acer truncatum extract is 8.0%.
The preparation method of the Acer truncatum extract comprises the following steps:
step 1: preparation of Acer acutangula crude extract
Drying stems, branches and leaves of the raw material Acer acutangum, and crushing to 2mm to obtain dried Acer acutangum powder.
Taking 600g of dried Acer acutus powder, adding trichloromethane 15 times of the mass of the Acer acutus powder, extracting under reflux at 60 ℃ for 3 times, each time for 1 hour, sieving with a 50-micron sieve, combining filtrates, and concentrating the filtrate under reduced pressure to an extract with a specific gravity of 1.05 at 65 ℃ to obtain 34.3g of Acer acutus crude extract.
Step 2: purification of
Purifying 30g of the Acer truncatum crude extract obtained in the step 1 by using a C18 column chromatography with the particle size of 40-60 mu m, removing impurities by using 40% methanol by volume concentration, eluting by using 75% methanol by volume concentration, collecting eluent, recovering solution, and drying to obtain 4.5g of purified Acer truncatum extract.
HPLC sample test solution preparation for this example: the same as in example 2.
Liquid chromatography conditions: the same as in example 1.
The results are shown in FIG. 3.
As can be seen from fig. 3, the content of carnosol was 59.6% by mass by HPLC analysis.
The Acer acutus extract is applied to the preparation of anti-inflammatory drugs or anti-inflammatory auxiliary drugs.
Example 4
The acer truncatum extract of the embodiment contains a mixture of carnosol, rosmanol and isorosmanol, wherein the mass content of the carnosol in the acer truncatum extract is 82.6%, the mass content of the rosmanol in the acer truncatum extract is 5.4%, and the mass content of the isorosmanol in the acer truncatum extract is 2.1%.
The preparation method of the Acer truncatum extract comprises the following steps:
step 1: preparation of Acer acutangula crude extract
Drying stems, branches and leaves of the raw material Acer acutangum, and crushing to 2mm to obtain dried Acer acutangum powder.
Taking 600g of dried Acer acutangum powder, adding acetone 20 times of the weight of the Acer acutangum powder, extracting under reflux at 70 ℃ for 3 times, each time for 1 hour, sieving by a 70-micron sieve, combining filtrates, and concentrating the filtrate under reduced pressure to an extract with the specific gravity of 1.08 at 65 ℃ to obtain 51.4g of Acer acutangum crude extract.
Step 2: purification of
And (2) purifying 30g of the Acer truncatum crude extract obtained in the step (1) by MCI column chromatography with the particle size of 75-150 microns, removing impurities by using 45% ethanol in volume concentration, eluting by using 85% ethanol in volume concentration, collecting eluent, recovering solution, and drying to obtain 10.4g of purified Acer truncatum extract.
HPLC sample test solution preparation for this example: the same as in example 2.
Liquid chromatography conditions: the same as in example 1.
The results are shown in FIG. 4.
As can be seen from fig. 4, the content of carnosol was 82.6% by mass by HPLC analysis.
The Acer acutus extract is applied to the preparation of anti-inflammatory drugs or anti-inflammatory auxiliary drugs.
Example 5
The acer truncatum extract of the embodiment contains a mixture of carnosol, rosmanol and isorosmanol, wherein the mass content of the carnosol in the acer truncatum extract is 63.2%, the mass content of the rosmanol in the acer truncatum extract is 7.9%, and the mass content of the isorosmanol in the acer truncatum extract is 4.9%.
The preparation method of the Acer truncatum extract comprises the following steps:
step 1: preparation of Acer acutangula crude extract
Drying stems, branches and leaves of the raw material Acer acutangum, and crushing to 2mm to obtain dried Acer acutangum powder.
Taking 600g of dried Acer acutangum powder, adding ethyl acetate 20 times of the weight of the Acer acutangum powder, performing reflux extraction for 3 times at 55 ℃, each time for 1 hour, sieving by a 55-micron sieve, combining filtrates, and concentrating the filtrate under reduced pressure to an extract with the specific gravity of 1.02 at 65 ℃ to obtain 43.6g of Acer acutangum crude extract.
Step 2: purification of
And (2) purifying 30g of the Acer truncatum crude extract obtained in the step (1) by silica gel column chromatography, removing oil and fat components by using petroleum ether, eluting by using a mixed solvent of the petroleum ether and acetone in a volume ratio of 6:1, collecting eluent, recovering the solvent, and drying to obtain 9.3g of the purified Acer truncatum extract.
HPLC sample test solution preparation for this example: the same as in example 2.
Liquid chromatography conditions: the same as in example 1.
The results are shown in FIG. 5.
As can be seen from fig. 5, the mass percentage of carnosol was 63.2% by HPLC analysis.
The Acer acutus extract is applied to the preparation of anti-inflammatory drugs or anti-inflammatory auxiliary drugs.
Example 6
The acer truncatum extract of the embodiment contains a mixture of carnosol, rosmanol and isorosmanol, wherein the content of carnosol in the acer truncatum extract is 53.0% by mass, the content of rosmanol in the acer truncatum extract is 4.5% by mass, and the content of isorosmanol in the acer truncatum extract is 4.4% by mass.
The preparation method of the Acer truncatum extract comprises the following steps:
step 1: preparation of Acer acutangula crude extract
Drying stems, branches and leaves of the raw material Acer acutangum, and crushing to 2mm to obtain dried Acer acutangum powder.
Taking 600g of dried Acer acutangum powder, adding ethyl acetate 20 times of the weight of the Acer acutangum powder, performing reflux extraction for 3 times at 65 ℃, each time for 1 hour, sieving by a sieve of 65 mu m, combining filtrates, and concentrating the filtrate under reduced pressure to an extract with the specific gravity of 1.06 at 65 ℃ to obtain 44.6g of Acer acutangum crude extract.
Step 2: purification of
And (2) purifying 30g of the Acer truncatum crude extract obtained in the step (1) by silica gel column chromatography, removing oil and fat components by using petroleum ether, eluting by using a mixed solvent of the petroleum ether and acetone in a volume ratio of 4:1, collecting eluent, recovering the solvent, and drying to obtain 12.8g of the purified Acer truncatum extract.
HPLC sample test solution preparation for this example: the same as in example 2.
Liquid chromatography conditions: the same as in example 1.
The results are shown in FIG. 6.
As can be seen from fig. 6, the mass percentage of carnosol was 53.0% by HPLC analysis.
The Acer acutus extract is applied to the preparation of anti-inflammatory drugs or anti-inflammatory auxiliary drugs.
NO production inhibition experiment
Taking RAW264.7 cells in logarithmic growth phase, counting with a cell counter, and making into 5 × 104-6×104The single cell suspension was inoculated into 12-well plates (1 mL/well) at 37 ℃ with 5% CO2Cultured in a cell culture box. After 24h of incubation, the drug-adding group (Acer acutus extract prepared in examples 2-6), LPS group (concentration of 0.2. mu.g/mL) and blank control group were set separately, wherein the drug-adding group was added with drugs (25. mu.g/mL, 12.5. mu.g/mL, 6.25. mu.g/mL, 3.125. mu.g/mL (25. mu.g/mL is the maximum safe concentration) and LPS with concentration of 0.2. mu.g/mL, and each group was set with 3 parallel wells. And after continuously culturing for 24 hours, collecting the culture medium, detecting the NO content in the culture medium by a Griess method, and calculating the IC50 value. The results are shown in Table 1.
TABLE 1 Effect of Acer acutus extracts prepared in examples 2-6 on NO release from LPS-induced RAW264.7 cells
As can be seen from table 1, the acer truncatum extracts prepared in examples 2-6 all showed strong NO inhibitory activity at safe concentrations. Therefore, the Acer acutangum extract can be applied to preparation of anti-inflammatory drugs or anti-inflammatory auxiliary drugs.
Mouse ear swelling test
70 KM male mice are selected, and the physical quality is 18-22 g. After 1 week of adaptive feeding (at this time, the body mass is 27-30g), the groups were randomly divided into 7 groups according to body mass, and a model group, a positive control group (aspirin) and an experimental group (acer truncatum extract prepared in examples 2-6) were set. After grouping, the medicine is administrated by intragastric administration according to 0.2mL/10g, the positive control group is administrated with 200mg/Kg aspirin, the experimental group is correspondingly administrated with 400mg/Kg of medicine, and the model group is administrated with the same amount of distilled water. And (3) performing intragastric administration for 1 time/d continuously for 1 hour after the last intragastric administration, and uniformly smearing 0.015mL of dimethylbenzene in front of and behind the right ear of 7 groups of mice by using a liquid-transferring gun. After 1h of smearing, the mice are killed by dislocation of cervical vertebrae, left and right ears are cut along the base line of auricles, round ear pieces are respectively punched at the same positions of the two ears by a 7mm puncher, weighing is carried out, and the swelling inhibition rate is calculated.
The results are shown in Table 2.
TABLE 2 Effect of Acer truncatum extract on ear swelling in mice
As can be seen from Table 2, the swelling degree of the model group reaches 15.64mg, the inhibition rate of the positive aspirin group reaches 33.76%, and the model is good. The acer acutissima extracts prepared in examples 2 to 6 all showed swelling inhibitory activity, as compared to the model group.
Therefore, the Acer acutangum extract disclosed by the invention is rich in diterpene phenol components, the components have various activities such as better oxidation resistance, cancer resistance and anti-inflammation, and a wider space is provided for further application of the Acer acutangum extract through pharmacodynamic study on the Acer acutangum extract. The Acer acutangum extract can be applied to preparing anti-inflammatory drugs or anti-inflammatory auxiliary drugs, and has wide application space.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (6)
1. The Acer acutangum extract is characterized by comprising a mixture of carnosol, rosmanol and isorosmanol, wherein the mass percent of the carnosol in the Acer acutangum extract is 53.0-82.6%, the mass percent of the rosmanol in the Acer acutangum extract is 4.5-12.4%, and the mass percent of the isorosmanol in the Acer acutangum extract is 2.1-8.0%; the preparation method of the Acer truncatum bunge extract comprises the following steps:
step 1: preparation of Acer acutangula crude extract
Taking dried Acer acutus power, adding an organic solvent which is 10-20 times of the weight of the Acer acutus power, extracting for 3 times under reflux, wherein the organic solvent is one of dichloromethane, chloroform, ethyl acetate and acetone, filtering, combining filtrates, and concentrating the filtrate under reduced pressure to obtain an extract, namely the Acer acutus crude extract;
step 2: purification of
Purifying the Acer truncatum crude extract obtained in the step 1 by silica gel column chromatography, removing oil and fat components by using petroleum ether, eluting by using a mixed solvent of the petroleum ether and ethyl acetate with a volume ratio of (5-3) to 1 or a mixed solvent of the petroleum ether and acetone with a volume ratio of (6-4) to 1, collecting eluent, recovering the solvent, and drying to obtain a purified Acer truncatum extract;
or purifying the Acer truncatum crude extract obtained in the step 1 by using reverse phase material column chromatography, wherein the medium of the reverse phase column chromatography is one of C18, C8, C4 or MCI, impurities are removed by using alcohol with the volume concentration of 40-50%, then the alcohol with the volume concentration of 75-95% is used for elution, eluent is collected, solution is recovered, and the purified Acer truncatum extract is obtained after drying, wherein the alcohol is methanol or ethanol.
2. The Acer acutangum extract as claimed in claim 1, wherein the Acer acutangum powder in step 1 is prepared by drying stems, branches and leaves of Acer acutangum, and pulverizing to 2 mm.
3. The Acer truncatum extract of claim 1, wherein the temperature of the extraction in step 1 is 50-70 ℃ for 1 hour each.
4. The Acer truncatum extract of claim 1, wherein the pore size of the filtration in step 1 is 30-70 μm.
5. The Acer truncatum bunge extract of claim 1, wherein the specific gravity of the extract in the step 1 at 65 ℃ is 1.02-1.08.
6. Use of the Acer acutus extract of claim 1 in the preparation of an anti-inflammatory agent or an anti-inflammatory adjuvant.
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| CN106266262A (en) * | 2015-06-05 | 2017-01-04 | 九芝堂股份有限公司 | A kind of Callicarpa nudiflora extract with antiinflammatory action |
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| JP2014097945A (en) * | 2012-11-14 | 2014-05-29 | Momotani Juntendo:Kk | Tyrosinase induction inhibitors and melanin synthesis inhibitors |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106266262A (en) * | 2015-06-05 | 2017-01-04 | 九芝堂股份有限公司 | A kind of Callicarpa nudiflora extract with antiinflammatory action |
Non-Patent Citations (4)
| Title |
|---|
| Callicarpa longissima extract，carnosol-rich，potently inhibits melanogenesis in B16F10 melanoma cells;Minori Yamahara，等;《Journal of Natural Medicines》;20160131;第70卷(第1期);摘要 * |
| 尖尾枫化学成分研究;高微，等;《中国实验方剂学杂志》;20131031;第19卷(第19期);第154页左栏第2-4段 * |
| 尖尾枫挥发油化学成分分析研究;高微，等;《香料香精化妆品》;20150630(第3期);正文第一段 * |
| 鼠尾草酚对人脐静脉内皮细胞黏附淋巴细胞能力的影响及其机制;姚晖，等;《山东医药》;20171231;第57卷(第23期);正文第一段 * |
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