CN110508031A - A method of separating ursolic acid and oleanolic acid from plant - Google Patents
A method of separating ursolic acid and oleanolic acid from plant Download PDFInfo
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- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
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- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/18—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns
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Abstract
A method of separating ursolic acid and oleanolic acid from plant, the method are as follows: the mixed solvent system of n-hexane, methylene chloride, first alcohol and water composition is added in separatory funnel, stratification after shake well, take and be added to acidic materials as stationary phase, under be added to alkaline matter as mobile phase；The mixed solution for the lower phase for taking the crude extract of plant stationary phase and not alkalizing dissolves, and obtains sample solution；Stationary phase is taken to fill the splitter of counter-current chromatograph, it is subsequently injected into sample solution, after the completion of sample introduction, opening speed controller rotates forward splitter, persistently inject mobile phase, UV detector detection collects eluent with automatic fraction collector, merges the eluent containing target substance, evaporating solvent under reduced pressure and drying, obtain target product；The present invention passes through pH zone refining countercurrent separating ursolic acid and oleanolic acid, simple process, rate of recovery height.
Description
(1) technical field
The method of the present invention relates to a kind of from plant separating ursolic acid and oleanolic acid.
(2) background technique
Ursolic acid is to be present in one of natural plants triterpene compound, has calm, anti-inflammatory, antibacterial, anti-glycosuria
Various biologicals effect, the ursolic acid such as disease, antiulcer, reduction blood glucose also have apparent anti-oxidation function, thus by widely
As medicine and cosmetic material.Ursolic acid clinical manifestation has significant and reduces rapidly glutamic-pyruvic transaminase, serum transaminase, recession
Yellow subcutaneous ulcer improves a poor appetite, the effect of anti-fibrosis and recovery liver function, has the characteristics that quick, short treating period, effect stability.Together
Pier tartaric acid exists very extensively in plant, and general content 0.2%~2% is broad spectrum antibiotic, clinically drops enzyme for protect liver,
Treat bronchitis, pneumonia, acute tonsillitis, periodontitis, bacillary dysentery, acute gastroenteritis, urinary system infection contamination.In addition, clinical
On be also used to treat oxyhepatitis.
Aralia wood (Aralia chinensis), is Umbellales Araliaceae, has dispelling wind and eliminating dampness, inducing diuresis to remove edema, promoting blood circulation
The effects of analgesic, can be used for hepatitis, enlargement of lymph nodes, nephritic dropsy, diabetes, stomachache, rheumatic arthritis, pain in waist and lower extremities, bruise
The treatment of the diseases such as damage.Apple skin is the pericarp of rosaceae Malus apple (Malus domestica), in apple skin
Antioxidant content and bioactive substance rich in, it is beneficial to health to eat apple skin.Loguat leaf is that rosaceae loquat belongs to plant
The effect of dried leaf of object loquat (Eriobotrya japonica Thunb.) has a clearing lung and relieving cough, stomach function regulating diuresis, quenches the thirst and
There are treatment lung feverish cough, hemoptysis, bleeding from five sense organs or subcutaneous tissue, stomach hot vomiting effect.Main active in oleanolic acid Shi Aralia wood, ursolic acid with
Oleanolic acid is active constituent important in apple skin and loguat leaf.
The Aralia wood whole nation is widely distributed, early very high by the civil content using oleanolic acid in , Aralia wood in various regions as medicinal material,
In development of the West Regions, conceding the land to forestry, afforestation, this resource of active development is answered.China is the world's largest apple production
State and country of consumption can generate a large amount of apple skin in current consumption and processing apple.If apple skin is not handled well, will produce
Raw serious problem of environmental pollution.Ursolic acid is the most abundant pentacyclic triterpenoid of content in apple skin, is become in recent years
Research hotspot.China is that all there are plantation, therefore loguat leaf in most important loguat leaf producing country, each province on the south the Changjiang river in the world
Resource is very rich.However, most of loguat leaves are still used as garbage disposal, if dealt with improperly, such as burn, can also pollute ring
Border.The content of ursolic acid and oleanolic acid is higher in loguat leaf, is extracted by the deep processing of the effective active composition to loguat leaf
Purifying research makes its " turning waste into wealth ", then its market prospects is inevitable very optimistic.Therefore, Devoting Major Efforts To Developing rich in ursolic acid and
The plant of oleanolic acid, by with good economic efficiency and social effect.
Currently, the extraction of ursolic acid and oleanolic acid mainly passes through silica gel column chromatography, silica gel column chromatography is cumbersome, step
More, process is slowly increased the production cycle, and obtaining object ursolic acid and oleanolic acid, time-consuming, at high cost, it is difficult to obtain wide
General application.PH zone refining countercurrent (pH-zone-refining countercurrent chromatography) be by
A kind of special separation technology that common adverse current chromatogram develops has sample volume big (reaching gram-grade), separation purity height, separation
The advantages that time is short.The prominent feature of the technology is to joined reservation acid or alkali in stationary phase, joined and washes in mobile phase
Dealkalize or acid, according to the dissociation constant (pK of separate substancea) and hydrophobic difference and realize separation, target components are with rectangle peak
It is eluted out, and impurity is then enriched in rectangle peak edge, and different component is eluted out the variation along with pH value.
It is mainly used in the preparatives such as organic acids and base derivative, polypeptide, organic dyestuff, alkaloid, isomer, free peptide point
From.
(3) summary of the invention
The object of the present invention is to provide the methods of a kind of preparative separating ursolic acid and oleanolic acid, rapidly by ursolic acid
It is separated from plant with two kinds of substances of oleanolic acid, simple process, there is stronger purpose, quickly and efficiently.
Technical scheme is as follows:
A method of separating ursolic acid and oleanolic acid, the method from plant are as follows:
(1) mixed solvent system of n-hexane, methylene chloride, first alcohol and water composition is added in separatory funnel, sufficiently shakes
Shake rear stratification, take and be added to acidic materials as stationary phase, under be added to alkaline matter as mobile phase；
In the mixed solvent system, n-hexane, methylene chloride, methanol, water volume parts be respectively 6~8 parts, 3~5
Part, 2~4 parts, 6~8 parts, preferably respectively 7 parts, 3 parts, 2 parts, 8 parts；
The acidic materials are preferably trifluoroacetic acid or acetic acid, most preferably trifluoroacetic acid, and the acidic materials are in stationary phase
In concentration be 5~20mM, preferably 10mM；
The alkaline matter is preferably ammonium hydroxide (25~28wt%NH3) or triethylamine, most preferably ammonium hydroxide, the alkaline matter
Concentration in mobile phase is 5~20mM, preferably 10mM；
(2) mixed solution for the lower phase for taking the crude extract of plant stationary phase and not alkalizing dissolves, and obtains sample solution；
The plant is, for example: Aralia wood, apple skin, loguat leaf；
The volume ratio of the stationary phase and the lower phase not alkalized is 1:1；
The volumetric usage of the mixed solution of the stationary phase and the lower phase not alkalized is calculated as with the quality of the crude extract of plant
0.1mL/mg；
(3) it takes stationary phase to fill the splitter of counter-current chromatograph, is subsequently injected into sample solution, after the completion of sample introduction, open
Speed control rotates forward splitter, under 550~850r/min (preferably 600r/min) revolving speed, (preferably with 1~2mL/min
Flow velocity 1.5mL/min) persistently injects mobile phase, is detected with the UV detector of 200~254nm of wavelength, is received with automatic part
Storage collects eluent, merges the eluent containing target substance, evaporating solvent under reduced pressure and drying, obtains target product；
The sample solution can be injected by sample introduction circle or by being pumped into counter-current chromatograph；
The sampling volume of the sample solution is usually less than the 30% of splitter column volume, preferably 5%~10%；
The target substance can be monitored by thin-layer chromatography or high performance liquid chromatography, be merged and ursolic acid and olive
Sour standard items RfValue or the identical eluent containing ursolic acid and oleanolic acid of retention time；
The solvent of the thin-layer chromatography is that the mixing of hexamethylene, ethyl acetate and glacial acetic acid volume ratio 10:3:0.5 is molten
Agent；
The testing conditions of the high performance liquid chromatography are as follows: 5 HC-C of Agilent18(2) column (4.6 × 250mm, 5 μ
m)；25 DEG C of column temperature；Mobile phase is -0.5% ammonium acetate solution of acetonitrile-methanol (67:12:21, v/v/v), isocratic elution mode；Stream
Speed: 1mL/min；Detection wavelength: 210nm；Sample volume: 20 μ L.
In the present invention, the preparation method of the crude extract of the plant enumerates as follows:
The crude extract of Aralia wood is made as follows:
Qu Aralia wood root powder, with 95% ethanol water refluxing extraction of volume fraction 2 times, 1 hour every time, extracting solution was evaporated off molten
Agent adds 10wt%H at thick medicinal extract shape, thick paste object2SO4Aqueous solution is condensed back hydrolysis 5h, after the completion of hydrolysis, by water at 100 DEG C
Sediment filtering is solved, then sediment is washed till neutrality with water repeatedly, sediment adds 10wt%NaOH aqueous solution, pH to 8~9 is adjusted,
70 DEG C are heated to, is filtered while hot, filtrate adds 10wt%HCl aqueous solution tune pH to 1, stands 8~12h, filters, the sediment filtered out
Dry get Dao Aralia wood crude extract.
The crude extract of apple skin is made as follows:
Apple skin powder is added in 100 DEG C of water and decocts 10min, repeats to decoct 5~7 times, is clarified until the water of filtering becomes,
Apple skin powder after decoction is placed on (40 DEG C) dryings of air -oven, is then mentioned with the reflux of 95% ethanol water of volume fraction
It takes 2~3 times, 1 hour every time, after extracting solution concentration, 10wt%HCl aqueous solution is added, adjusts pH to 3, stands 8~12h, filters,
Filter cake petroleum ether, drying obtain the crude extract of apple skin.
The crude extract of loguat leaf is made as follows:
Dry loguat leaf is crushed, 20 meshes is crossed, weighs loguat leaf fine powder, 95% ethanol water of volume fraction is added to condense
Refluxing extraction 1h, filtering, filter residue repeat refluxing extraction 2~3 times, combined extract, and active carbon is added and is warming up to 70 DEG C, oscillation is de-
Color 20min, filtering, filtrate decompression are concentrated to dryness, and obtain the crude extract of loguat leaf.
The beneficial effects of the present invention are: pass through pH zone refining countercurrent separating ursolic acid and oleanolic acid, technique
Simply, the rate of recovery is high, provides new approaches for ursolic acid and isolating and purifying for oleanolic acid.
(4) Detailed description of the invention
Fig. 1: high speed adverse current chromatogram (HSCCC) figure of embodiment 1.
Fig. 2: high speed adverse current chromatogram (HSCCC) figure of embodiment 2.
Fig. 3: high speed adverse current chromatogram (HSCCC) figure of embodiment 3.
(5) specific embodiment
Technical solution of the present invention is described further with specific embodiment below, but protection scope of the present invention is not
It is limited to this.
Semi-preparative adverse current chromatogram is used in the embodiment of the present invention, (MP0106, Shanghai three are counter-current chromatograph by constant flow pump
Scientific instrument Co., Ltd), UV detector (UVD-680-3, Shanghai Jin Da biochemical instrument Co., Ltd), recorder (Jin Da
Work station, Shanghai Jin Da biochemical instrument Co., Ltd) etc. composition.
Embodiment 1:
Drying Aralia wood is crushed, 20 mesh , Qu Aralia wood powder 50g are crossed, 95% alcohol reflux measured with 10 times extracts 2 times
(every time 1 hour), filtering recycle ethyl alcohol into thick medicinal extract shape, and thick paste object adds the 10%H of 100mL2SO4It is condensed back at 100 DEG C
5h is hydrolyzed, after the completion of hydrolysis, hydrolytic precipitation object is filtered, then precipitating is washed till neutrality with water repeatedly, sediment adds 10%NaOH
Solution is adjusted pH to 8-9, is filtered while hot after being heated to 70 DEG C.Filtrate adds 10% hydrochloric acid tune pH to 1, and after standing overnight, drying is heavy
It forms sediment, obtains crude product 1.014g.
N-hexane, methylene chloride, first alcohol and water are configured in separatory funnel according to volume ratio 7:3:2:8, shake well
Stratification afterwards is above added to trifluoroacetic acid, makes the final concentration of 10mM of trifluoroacetic acid as stationary phase, under be added to ammonium hydroxide
(25%~28%NH3), make the final concentration of 10mM of ammonium hydroxide as mobile phase；Cheng Qu Aralia wood crude extract 100mg, is fixed with 5ml
The lower phased soln not alkalized with 5ml mutually, obtains sample solution.
Loquat Leave extract is separated using semi-preparative counter-current chromatograph, the column volume of splitter is 105ml.It is first
Stationary phase is first filled into splitter with the flow velocity of 10ml/min, above-mentioned sample solution is then injected into splitter by sample introduction circle,
After the completion of sample introduction, opening speed controller rotates forward splitter, and adjusting revolving speed is 600r/min, and the flow velocity of mobile phase is arranged
For 1.5ml/min, start to be pumped into mobile phase, receives eluent by the speed of 3 minutes/pipe with automatic fraction collector, use thin layer
(solvent is hexamethylene: ethyl acetate: glacial acetic acid=10:3:0.5) tracing detection is chromatographed into eluent without oleanolic acid group
Point, stop collecting.It collects obtained eluent to be detected by thin-layer chromatography and efficient liquid phase, merge and oleanolic acid standard
Product RfIt is worth (Rf=0.67) and the identical eluent of retention time (21.7min) with Rotary Evaporators recycling design obtains olive
Sour 38.56mg, the wherein purity 99.01% of oleanolic acid, the rate of recovery 90.23%.
The testing conditions of high performance liquid chromatography are as follows: 5 HC-C of Agilent18(2) column (4.6 × 250mm, 5 μm)；Column
25 DEG C of temperature；Mobile phase is -0.5% ammonium acetate solution of acetonitrile-methanol (67:12:21, v/v/v), isocratic elution mode；Detect wave
It is long: 210nm；Sample volume: 20 μ L.
Embodiment 2:
Dry apple skin is crushed, 20 meshes is crossed, weighs 50g apple skin powder, be added in 100 DEG C of water and decoct
10min is repeated several times (5-7 times), clarifies until remaining water becomes, places it in (40 DEG C) dryings of air -oven, is then added
10 times of 95% ethanol solutions of amount are condensed back 1h, filtering, and 95% ethanol solution that residue is measured with 10 times repeats above-mentioned reflux 2 times,
After concentrated extract to 50mL, 10% dilute hydrochloric acid is slowly added into concentrate, pH to 3 or so is adjusted, stood for merging filtrate
Night filters, and for filter cake with petroleum ether 2 times, drying obtains crude product 2.353g.
N-hexane, methylene chloride, first alcohol and water are configured in separatory funnel according to volume ratio 7:3:2:8, shake well
Stratification afterwards is above added to trifluoroacetic acid, makes the final concentration of 10mM of trifluoroacetic acid as stationary phase, under be added to ammonium hydroxide
(25%~28%NH3), make the final concentration of 10mM of ammonium hydroxide as mobile phase；Apple skin crude extract 100mg is weighed, it is solid with 5ml
The fixed lower phased soln not alkalized with 5ml mutually, obtains sample solution.
Loquat Leave extract is separated using semi-preparative counter-current chromatograph, the column volume of splitter is 105ml.It is first
Stationary phase is first filled into splitter with the flow velocity of 10ml/min, above-mentioned sample solution is then injected into splitter by sample introduction circle,
After the completion of sample introduction, opening speed controller rotates forward splitter, and adjusting revolving speed is 600r/min, and the flow velocity of mobile phase is arranged
For 1.5ml/min, start to be pumped into mobile phase, receives eluent by the speed of 3 minutes/pipe with automatic fraction collector, use thin layer
(solvent is hexamethylene: ethyl acetate: glacial acetic acid=10:3:0.5) tracing detection is chromatographed into eluent without ursolic acid and neat
Pier tartaric acid component stops collecting.It collects obtained eluent to be detected by thin-layer chromatography and efficient liquid phase, merging and black bearberry
Sour standard items RfIt is worth (Rf=0.60) and retention time (22.5min) is identical and oleanolic acid standard items RfIt is worth (Rf=0.67)
And the identical eluent of retention time (21.7min) obtains the mixing of ursolic acid and oleanolic acid with Rotary Evaporators recycling design
Object 65.6mg, the wherein purity 90.98% of ursolic acid, the rate of recovery 89.66%；The purity of oleanolic acid is 6.51%, recycling
Rate is 88.89%.
The testing conditions of high performance liquid chromatography are as follows: Agilent 5 HC-C18 (2) column (4.6 × 250mm, 5 μm)；Column
25 DEG C of temperature；Mobile phase is -0.5% ammonium acetate solution of acetonitrile-methanol (67:12:21, v/v/v), isocratic elution mode；Detect wave
It is long: 210nm；Sample volume: 20 μ L.
Embodiment 3:
Dry loguat leaf is crushed, 20 meshes is crossed, weighs the loguat leaf fine powder of 100g, add 10 times of 95% ethyl alcohol of amount molten
Liquid is condensed back 1h, filtering, and 95% ethanol solution that residue is measured with 10 times repeats above-mentioned reflux 2 times, and merging filtrate, filtrate adds
Enter 1.5% active carbon, 70 DEG C of decolorations are filtered after oscillation decoloration 20min, then concentrate filtrate to dry, obtain loguat leaf
Ethanol extract, obtain 1.535g crude extract.
N-hexane, methylene chloride, first alcohol and water are configured in separatory funnel according to volume ratio 7:3:2:8, shake well
Stratification afterwards is above added to trifluoroacetic acid, makes the final concentration of 10mM of trifluoroacetic acid as stationary phase, under be added to ammonium hydroxide
(25%~28%NH3), make the final concentration of 10mM of ammonium hydroxide as mobile phase；Loquat Leave extract 100mg is weighed, it is solid with 5ml
The fixed lower phased soln not alkalized with 5ml mutually, obtains sample solution.
Loquat Leave extract is separated using semi-preparative counter-current chromatograph, the column volume of splitter is 105ml.It is first
Stationary phase is first filled into splitter with the flow velocity of 10ml/min, above-mentioned sample solution is then injected into splitter by sample introduction circle,
After the completion of sample introduction, opening speed controller rotates forward splitter, and adjusting revolving speed is 600r/min, and the flow velocity of mobile phase is arranged
For 1.5ml/min, start to be pumped into mobile phase, receives eluent by the speed of 3 minutes/pipe with automatic fraction collector, use thin layer
(solvent is hexamethylene: ethyl acetate: glacial acetic acid=10:3:0.5) tracing detection is chromatographed into eluent without ursolic acid and neat
Pier tartaric acid component stops collecting.It collects obtained eluent to be detected by thin-layer chromatography and efficient liquid phase, merging and black bearberry
Sour standard items RfIt is worth (Rf=0.60) and retention time (22.5min) is identical and oleanolic acid standard items RfIt is worth (Rf=0.67)
And the identical eluent of retention time (21.7min) obtains the mixing of ursolic acid and oleanolic acid with Rotary Evaporators recycling design
Object 46.6mg, the wherein purity 74.35% of ursolic acid, the rate of recovery 86.59%；The purity of oleanolic acid is 23.61%, recycling
Rate is 90.99%.
The testing conditions of high performance liquid chromatography are as follows: 5 HC-C of Agilent18(2) column (4.6 × 250mm, 5 μm)；Column
25 DEG C of temperature；Mobile phase is -0.5% ammonium acetate solution of acetonitrile-methanol (67:12:21, v/v/v), isocratic elution mode；Detect wave
It is long: 210nm；Sample volume: 20 μ L.
Comparative example:
(Niu Huiying vinca alkaloids extract the separation purifying technique of ursolic acid and oleanolic acid in residue to Niu Huiying
Study [D] Northeast Forestry University, 2008.) ursolic acid and neat is extracted in residue using column chromatography purifying vinca alkaloids
Pier tartaric acid, specific process parameter are as follows: column chromatographic stuffing is 300-400 mesh silica gel, and applied sample amount is 0.2g/10g silica gel, is eluted molten
Agent is n-hexane: ethyl acetate=5:4, flow velocity 3ml/min, collects corresponding outflow component.4.84g vinca alkaloids acetic acid
Sample 2.28g is obtained after ethyl ester extract column chromatography, the purity of ursolic acid is 68.14% in column chromatographed product, and the rate of recovery is
86.05%；The purity of oleanolic acid is 17.79%, the rate of recovery 81.21%.
The production cost of the method is relatively high, the technique very complicated and production cycle is long, is not suitable for large-scale production.And
It is easy to operate using the ursolic acid and oleanolic acid in pH zone refining countercurrent separation loguat leaf in the present invention, when separation
Between it is short, the rate of recovery is high, and obtained ursolic acid and oleanolic acid purity is high has certain promotional value.
Claims (9)
1. a kind of method of the separating ursolic acid from plant and oleanolic acid, which is characterized in that the method are as follows:
(1) mixed solvent system of n-hexane, methylene chloride, first alcohol and water composition is added in separatory funnel, after shake well
Stratification takes and is added to acidic materials as stationary phase, under be added to alkaline matter as mobile phase；
In the mixed solvent system, n-hexane, methylene chloride, methanol, water volume parts be respectively 6~8 parts, 3~5 parts, 2
~4 parts, 6~8 parts；
The acidic materials are trifluoroacetic acid or acetic acid, and concentration of the acidic materials in stationary phase is 5~20mM；
The alkaline matter is ammonium hydroxide or triethylamine, and concentration of the alkaline matter in mobile phase is 5~20mM；
(2) mixed solution for the lower phase for taking the crude extract of plant stationary phase and not alkalizing dissolves, and obtains sample solution；
(3) it takes stationary phase to fill the splitter of counter-current chromatograph, is subsequently injected into sample solution, after the completion of sample introduction, opening speed
Controller rotates forward splitter, under 550~850r/min revolving speed, persistently injects mobile phase with the flow velocity of 1~2mL/min, with
The UV detector of 200~254nm of wavelength detects, and collects eluent with automatic fraction collector, merges washing containing target substance
De- liquid, evaporating solvent under reduced pressure and drying, obtain target product.
2. as described in claim 1 from plant separating ursolic acid and oleanolic acid method, which is characterized in that step (1)
In, in the mixed solvent system, n-hexane, methylene chloride, methanol, water volume parts be respectively 7 parts, 3 parts, 2 parts, 8 parts.
3. as described in claim 1 from plant separating ursolic acid and oleanolic acid method, which is characterized in that step (2)
In, the plant is: Aralia wood, apple skin, loguat leaf.
4. as described in claim 1 from plant separating ursolic acid and oleanolic acid method, which is characterized in that step (2)
In, the volume ratio of the stationary phase and the lower phase not alkalized is 1:1.
5. as described in claim 1 from plant separating ursolic acid and oleanolic acid method, which is characterized in that step (2)
In, the volumetric usage of the mixed solution of the stationary phase and the lower phase not alkalized is calculated as 0.1mL/ with the quality of the crude extract of plant
mg。
6. as described in claim 1 from plant separating ursolic acid and oleanolic acid method, which is characterized in that step (3)
In, the sample solution is injected by sample introduction circle or by being pumped into counter-current chromatograph.
7. as described in claim 1 from plant separating ursolic acid and oleanolic acid method, which is characterized in that step (3)
In, the sampling volume of the sample solution is less than the 30% of splitter column volume.
8. as described in claim 1 from plant separating ursolic acid and oleanolic acid method, which is characterized in that step (3)
In, the target substance is monitored by thin-layer chromatography or high performance liquid chromatography, is merged and ursolic acid and oleanolic acid standard
Product RfValue or the identical eluent containing ursolic acid and oleanolic acid of retention time；
The solvent of the thin-layer chromatography is the mixed solvent of hexamethylene, ethyl acetate and glacial acetic acid volume ratio 10:3:0.5；
The testing conditions of the high performance liquid chromatography are as follows: 5 HC-C of Agilent18(2)column；25 DEG C of column temperature；Mobile phase is second
The mixed solution of -0.5% ammonium acetate solution volume ratio 67:12:21 of nitrile-methanol, isocratic elution mode；Flow velocity: 1mL/min；Inspection
Survey wavelength: 210nm；Sample volume: 20 μ L.
9. as described in claim 1 from plant separating ursolic acid and oleanolic acid method, which is characterized in that the plant
Crude extract Wei Aralia wood, apple skin or loguat leaf crude extract, the crude extract of the plant the preparation method comprises the following steps:
The crude extract of Aralia wood is made as follows:
Qu Aralia wood root powder, with 95% ethanol water refluxing extraction of volume fraction 2 times, every time 1 hour, extracting solution be evaporated off solvent at
Thick medicinal extract shape, thick paste object add 10wt%H2SO4Aqueous solution is condensed back hydrolysis 5h at 100 DEG C, after the completion of hydrolysis, hydrolysis is heavy
Starch filtering, then sediment is washed till neutrality with water repeatedly, sediment adds 10wt%NaOH aqueous solution, adjusts pH to 8~9, heating
It to 70 DEG C, filters while hot, filtrate adds 10wt%HCl aqueous solution tune pH to 1, stands 8~12h, filters, the sediment drying filtered out
Crude extract of the get Dao Aralia wood；
The crude extract of apple skin is made as follows:
Apple skin powder is added in 100 DEG C of water and decocts 10min, repeats to decoct 5~7 times, clarifies, will decoct until the water of filtering becomes
Apple skin powder after boiling is placed on air -oven drying, then uses 95% ethanol water refluxing extraction of volume fraction 2~3 times,
1 hour every time, after extracting solution concentration, 10wt%HCl aqueous solution is added, adjusts pH to 3, stands 8~12h, filters, filter cake petroleum
Ether washing, drying obtain the crude extract of apple skin；
The crude extract of loguat leaf is made as follows:
Dry loguat leaf is crushed, 20 meshes is crossed, weighs loguat leaf fine powder, 95% ethanol water of volume fraction is added to be condensed back
1h, filtering are extracted, filter residue repeats refluxing extraction 2~3 times, combined extract, and active carbon is added and is warming up to 70 DEG C, oscillation decoloration
20min, filtering, filtrate decompression are concentrated to dryness, and obtain the crude extract of loguat leaf.
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