CN107029028A - A kind of application of leaflet bush cinqefoil extract and preparation method and its antitumor activity - Google Patents
A kind of application of leaflet bush cinqefoil extract and preparation method and its antitumor activity Download PDFInfo
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Abstract
Application the invention discloses a kind of leaflet bush cinqefoil extract and preparation method thereof and its in antitumor activity, step is as follows：（1）It dry, pulverize；（2）Sub-critical extraction；（3）Microfiltration membranes, milipore filter and nanofiltration membrane treatment；（4）Freeze-drying, obtains a kind of leaflet bush cinqefoil extract with obvious antitumor activity.The inventive method not only can effectively prepare a kind of leaflet bush cinqefoil extract, especially preparation technology is very efficient, processing step mechanization degree is high, and leaflet bush cinqefoil extract made from the invention has the activity for significantly suppressing human nerve cell oncocyte SH SY5Y cells propagation, is a kind of antitumor agent of great application prospect.
Description
Technical field
The present invention relates to biological technical field, more particularly to a kind of leaflet bush cinqefoil extract and preparation method and its anti-swollen
The application of tumor activity.
Background technology
Leaflet bush cinqefoil（Potentilla parvifolia Fisch.）, also known as gold Lao Mei medicines, littleleaf ciquefoil leaf and flower, Ban Na
With, Bai La, Banma etc., rose family Potentilla machaka, be distributed widely in Sichuan of China, the Inner Mongol, Xinjiang, Qinghai,
The ground such as Gansu, Shanxi, are a kind of traditional middle Tibetan medicines.
《Jingzhubencao》Record：Ban Na closes treatment female breast distending pain.《Blue coloured glaze》Record：Ban Na closes stem red, spends Huang
Color, leaf is small, ashing, treats female breast distending pain, treating ascites.Bush cinqefoil rhizome, leaf, flower and branch have very strong pharmacology to live
Property, leaf and flower can be used as medicine, and slight bitter is cold in nature, clear away heart-fire with clearing away summerheat, stomach invigorating, invigorating the spleen, brain tonic, treat female irregular menstruation, solid
The functions such as tooth, rhizome extract has enhanced immunomodulatory properties, the extract edible of branch, moreover it is possible to added as food
Cholesterol and blood-sugar content in agent, reduction blood.Leaf among the people with bush cinqefoil is used on behalf of the tea, can effectively prevent heart and brain
Vascular diseases.《Dictionary of medicinal plant》Described in littleleaf ciquefoil leaf and flower major function：Diuresis disappears water, controls the disturbance of lower legs due to pathogenic cold and dampness, pruigo, mammary gland
It is scorching.
The various plants of Potentilla are medicinal plant, and the main component of the platymiscium has flavonoids, triterpenes, tannin
Class compound etc., with anti-inflammatory, anti-diabetic is antitumor, anti-oxidant, the multiple pharmacological effect such as anti-bacteria and anti-virus.
At present, the modern pharmacology research to leaflet bush cinqefoil is less, is badly in need of providing a kind of carrying with antitumor activity
Take thing and preparation method, and its antitumor activity application.
The content of the invention
Medicinal material used of the invention is leaflet bush cinqefoil（Potentilla parvifolia Fisch.）Drying overground part
Point, adopted in August, 2014 in Haibei Prefecture, plant specimen is stored in Qinghai-Tibet biological sample shop, sample numbering is
252558。
It is an object of the present invention to provide a kind of preparation method of leaflet bush cinqefoil extract.
The preparation method for the leaflet bush cinqefoil extract that the present invention is provided, comprises the following steps：
（1）By leaflet bush cinqefoil, dry, shred, obtain leaflet bush cinqefoil powder；
（2）Leaflet bush cinqefoil powder is placed in sub-critical extraction post（5L）In, ultra-pure water is conveyed and preheated into preheater by compression pump,
Reach after design temperature, be pumped into extraction column, system pressure is reached after setting value, and extraction column starts timing when filling water, extraction
Extract solution flow to collection kettle through cooler and collected after certain time, obtains leaflet bush cinqefoil subcritical abstraction liquid；
（3）Subcritical abstraction liquid is used into microfiltration membranes, milipore filter and nanofiltration membrane treatment successively, microfiltration membranes permeate, ultrafiltration are obtained successively
Film permeate and NF membrane retention component；
（4）By NF membrane retention component freeze-drying, a kind of leaflet bush cinqefoil extract is obtained.
Step（1）In, 0.4 ~ 1.0cm of leaflet bush cinqefoil powder path length.
Step（2）In, the subcritical abstraction agent is water, and temperature is 140 DEG C ~ 160 DEG C, and 4 ~ 6MPa of extracting pressure is extracted
Time is 10min ~ 40min, solid-liquid ratio 1:10g/mL ~1:30g/mL, the number of times of extraction is 2 ~ 4 times.
Step（3）In, the micro-filtration membrane aperture that micro-filtration film process is used is 0.02 μm ~ 8.00 μm, 20 DEG C of the temperature of processing, pressure
Power is 0.1 ~ 0.3MPa, and flow velocity is 10 ~ 30kg/h, obtains microfiltration membranes permeate.
Step（3）In, the ultrafiltration membrane aperture that ultrafiltration membrane treatment is used is 0.003 μm ~ 0.090 μm, 20 DEG C of temperature, pressure
0.1 ~ 0.3MPa, flow velocity is 10 ~ 30kg/h, obtains milipore filter permeate.
Step（3）In, the nanofiltration membrane aperture that nanofiltration membrane treatment is used is 0.001 μm ~ 0.002 μm, 20 DEG C of temperature, pressure
0.7 ~ 0.9MPa, 30 ~ 50kg/h of flow velocity, molecular cut off are 100 ~ 2000 leaflet bush cinqefoil extracted component, obtain NF membrane
Retain component.
Step（4）In, the freeze-drying program is -60 DEG C ~ -80 DEG C 1 ~ 3h of pre-freeze, -60 DEG C ~ -80 DEG C freeze-dryings 12
~ 36h, 5 ~ 10Pa of vacuum.
Step（4）In, in described leaflet bush cinqefoil extract, terpenoid content 5.42% ~ 17.61%, flavonoids
Compounds content 7.35% ~ 30.82%.
It is a further object to provide a kind of antitumor activity agent.
The antitumor activity agent that the present invention is provided, its active component is above-mentioned leaflet bush cinqefoil extract.
The experiment proves that, the present invention has advantages below compared with prior art：
（1）The present invention can be produced from leaflet bush cinqefoil to be obtained a kind of to suppress carrying for human nerve cell oncocyte SH-SY5Y
Thing is taken, is a kind of pure natural, plant source and no dependence tumor inhibitor；
（2）The extract that the present invention is obtained can significantly suppress human neuroblastoma cells SH-SY5Y propagation, its activity
It is 0.074 times of broad spectrum activity tumor inhibitor medicine 5-Fluorourail, IC50Respectively 10.89 μ g/mL and 0.14 μ g/mL,
Show the ability of stronger suppression SH-SY5Y propagation；
（3）The preparation technology of present invention offer leaflet bush cinqefoil extract realizes the mechanization preparation flow of closed loop substantially, significantly
Extract preparation time has been saved, extract preparation efficiency is improved.
In a word, the present invention not only enriches the source of pure natural antitumor activity agent, also opened up with region feature and
The leaflet bush cinqefoil plant of resources advantage new effect and application value.
Experimental method used in following embodiments is conventional method unless otherwise specified.
Material, reagent used etc., unless otherwise specified, are commercially obtained in following embodiments.
The present invention is described in further details below by embodiment, but is not the limit to the present invention
System, according to the above of the present invention, according to the ordinary technical knowledge and customary means of this area, not departing from, the present invention is above-mentioned
Under the premise of basic fundamental thought, the modification of other diversified forms can also be made, replaces or changes.
Embodiment
Subcritical abstraction equipment, purchased from Changzhou Te Wei electrical automation systems Co., Ltd；Microfiltration membranes ~ milipore filter ~ are received
Filter membrane system, purchased from Hefei Yu Wangmo engineering technology Co., Ltd；Human nerve cell oncocyte SH-SY5Y, purchased from Shanghai cell
Storehouse.
Embodiment 1：The preparation method of leaflet bush cinqefoil extract and its antitumor activity application.
First, preparation method
（1）It by leaflet bush cinqefoil, dry, pulverize, obtain leaflet bush cinqefoil powder, the long 1.0cm of particle diameter；
（2）The kg of leaflet bush cinqefoil powder 1.0 is placed in sub-critical extraction post and extracted, extractant is water, temperature is 140 DEG C,
Extracting pressure 4MPa, extraction time is 10min, solid-liquid ratio 1:10g/mL, the number of times of extraction is 2 times；
（3）Subcritical abstraction liquid is used into microfiltration membranes, milipore filter and nanofiltration membrane treatment successively.Wherein, micro-filtration membrane aperture is 8.00 μm,
20 DEG C of temperature, pressure is 0.3MPa, and flow velocity is 10kg/h, removes the impurity such as oil removal, bacterium and macromolecule colloid, obtains micro-
Filter membrane permeate；Ultrafiltration membrane aperture is 0.090 μm, 20 DEG C of temperature, and pressure 0.3MPa, flow velocity is 10kg/h, removes colloid, albumen
Matter, microorganism and larger molecular organicses, obtain milipore filter permeate；Nanofiltration membrane aperture is 0.002 μm, 20 DEG C of temperature, pressure
0.9MPa, flow velocity 30kg/h, molecular cut off are 100 ~ 2000 leaflet bush cinqefoil extracted component, obtain NF membrane retention group
Point；
（4）By NF membrane retention component freeze-drying, freeze-drying condition is -60 DEG C of pre-freeze 1h, and -60 DEG C are freeze-dried 12h, very
Reciprocal of duty cycle 5Pa, obtains a kind of leaflet bush cinqefoil extract A 65.08g.
2nd, the detection of leaflet bush cinqefoil
（1）The measure of terpenoid content in leaflet bush cinqefoil extract：It is published in April, 2008 with reference to Yang Manqin etc.《Peace
Emblem college of traditional Chinese medicine journal》Volume 27 the 2nd phase article " the outer Fen light Guang degree methods Ce of Zi Ding Common St.Paulswort Herb in total ter penoidses content " record
Method, terpenoid content in leaflet bush cinqefoil is measured；
（2）Flavonoid content in leaflet bush cinqefoil extract：With reference to People's Republic of China (PRC) agricultural industry criteria NYT
1295-2007《The measure of general flavone content in buckwheat and its product》The method of page 1 ~ page 3 record, in leaflet bush cinqefoil
Flavonoid content is measured；
（3）Inhibitory action of the leaflet bush cinqefoil extract to human nerve cell oncocyte SH-SY5Y：Using mtt assay to leaflet gold
Reveal prunus mume extract to carry out suppressing the test of human neuroblastoma cells SH-SY5Y proliferation activities.Leaflet bush cinqefoil extract it is dense
Degree is respectively 0.00,6.25,12.50,25.00,50.00,100.00 μ g/mL, with every hole 104Individual cell is inoculated in 96 orifice plates
In, per the μ L of pore volume 200, culture plate is put into incubator, in 37 DEG C, 5% CO2Saturated humidity under the conditions of cultivate 12 h
Afterwards, add after given the test agent, given the test agent effect 24h, 10 μ L 5mg/mL MTT solution is added per hole, in 37 DEG C, 5%CO2
And 4h is incubated under the conditions of saturated humidity, discard culture medium and terminate culture, 100 μ L DMSO are added per hole, vibrate 10min, make first a ceremonial jade-ladle, used in libation
Fully dissolving.Light absorption value is determined at 490nm with multi-function microplate reader, leaflet bush cinqefoil extract is calculated to the half of SH-SY5Y
Inhibiting rate IC50, and be compared using antineoplastic 5-Fluorourail as positive control.
After testing, terpenoid content 5.42%, flavones in the leaflet bush cinqefoil extract that the inventive method is prepared
Kind compound content 17.35%, SH-SY5Y semi-inhibit rate IC50For 10.89 μ g/mL.In addition, positive control medicine 5-
Fluorourail SH-SY5Y semi-inhibit rates IC50For 0.14 μ g/mL.
Embodiment 2：The preparation method of leaflet bush cinqefoil extract and its antitumor activity application.
First, preparation method
（1）It by leaflet bush cinqefoil, dry, pulverize, obtain leaflet bush cinqefoil powder, the long 0.4cm of particle diameter；
（2）Leaflet bush cinqefoil powder 2.0kg is placed in sub-critical extraction post and extracted, extractant is water, temperature is 160 DEG C,
Extracting pressure 6MPa, extraction time is 40min, solid-liquid ratio 1:30g/mL, the number of times of extraction is 4 times；
（3）Subcritical abstraction liquid is used into microfiltration membranes, milipore filter and nanofiltration membrane treatment successively.Wherein, micro-filtration membrane aperture is 0.02 μm,
20 DEG C of temperature, pressure is 0.1MPa, and flow velocity is 30kg/h, removes the impurity such as oil removal, bacterium and macromolecule colloid, obtains micro-
Filter membrane permeate；Ultrafiltration membrane aperture is 0.003 μm, 20 DEG C of temperature, and pressure 0.1MPa, flow velocity is 30kg/h, removes colloid, albumen
Matter, microorganism and larger molecular organicses, obtain milipore filter permeate；Nanofiltration membrane aperture is 0.001 μm, 20 DEG C of temperature, pressure
0.7MPa, flow velocity 50kg/h, molecular cut off are 100 ~ 2000 leaflet bush cinqefoil extracted component, obtain NF membrane retention group
Point；
（4）By NF membrane retention component freeze-drying, freeze-drying condition is -80 DEG C of pre-freeze 3h, and -80 DEG C are freeze-dried 36h, very
Reciprocal of duty cycle 10Pa, obtains a kind of leaflet bush cinqefoil extract B 174.25g.
2nd, the detection of leaflet bush cinqefoil
（1）Terpenoid content in leaflet bush cinqefoil extract：With the two of embodiment 1（1）It is identical；
（2）Flavonoid content in leaflet bush cinqefoil extract：With the two of embodiment 1（2）It is identical；
（3）Inhibitory action of the leaflet bush cinqefoil extract to human nerve cell oncocyte SH-SY5Y：With the two of embodiment 1（3）
It is identical.
After testing, terpenoid content 14.61% in the leaflet bush cinqefoil extract that the inventive method is prepared, it is yellow
Ketone compounds content 30.82%, SH-SY5Y semi-inhibit rate IC50For 6.24 μ g/mL.
Embodiment 3：The preparation method of leaflet bush cinqefoil extract and its antitumor activity application.
First, preparation method
（1）It by leaflet bush cinqefoil, dry, pulverize, obtain leaflet bush cinqefoil powder, the long 0.7cm of particle diameter；
（2）Leaflet bush cinqefoil powder 4.0kg is placed in sub-critical extraction post and extracted, extractant is water, temperature is 150 DEG C,
Extracting pressure 5MPa, extraction time is 25min, solid-liquid ratio 1:20g/mL, the number of times of extraction is 3 times；
（3）Subcritical abstraction liquid is used into microfiltration membranes, milipore filter and nanofiltration membrane treatment successively.Wherein, micro-filtration membrane aperture is 1.00 μm,
20 DEG C of temperature, pressure is 0.2MPa, and flow velocity is 20kg/h, removes the impurity such as oil removal, bacterium and macromolecule colloid, obtains micro-
Filter membrane permeate；Ultrafiltration membrane aperture is 0.030 μm, 20 DEG C of temperature, and pressure 0.2MPa, flow velocity is 20kg/h, removes colloid, albumen
Matter, microorganism and larger molecular organicses, obtain milipore filter permeate；Nanofiltration membrane aperture is 0.002 μm, 20 DEG C of temperature, pressure
0.8MPa, flow velocity 40kg/h, molecular cut off are 100 ~ 2000 leaflet bush cinqefoil extracted component, obtain NF membrane retention group
Point；
（4）By NF membrane retention component freeze-drying, freeze-drying condition is -70 DEG C of pre-freeze 2h, and -70 DEG C are freeze-dried 24h, very
Reciprocal of duty cycle 7Pa, obtains a kind of leaflet bush cinqefoil extract C 362.74g.
2nd, the detection of leaflet bush cinqefoil extract
（1）Terpenoid content in leaflet bush cinqefoil extract：With the two of embodiment 1（1）It is identical；
（2）Flavonoid content in leaflet bush cinqefoil extract：With the two of embodiment 1（2）It is identical；
（3）Inhibitory action of the leaflet bush cinqefoil extract to human nerve cell oncocyte SH-SY5Y：With the two of embodiment 1（3）
It is identical.
After testing, terpenoid content 9.03%, flavones in the leaflet bush cinqefoil extract that the inventive method is prepared
Kind compound content 24.61%, SH-SY5Y semi-inhibit rate IC50For 8.51 μ g/mL.
Comparative example 1：The preparation method of leaflet bush cinqefoil extract and its antitumor activity application.
First, preparation method
（1）It by leaflet bush cinqefoil, dry, pulverize, obtain leaflet bush cinqefoil powder, the long 0.7cm of particle diameter；
（2）The kg of leaflet bush cinqefoil powder 1.0 is placed in sub-critical extraction post and extracted, extractant is water, temperature is 130 DEG C,
Extracting pressure 3MPa, extraction time is 25min, solid-liquid ratio 1:20g/mL, the number of times of extraction is 3 times；
（3）Subcritical abstraction liquid is used into microfiltration membranes, milipore filter and nanofiltration membrane treatment successively.Wherein, micro-filtration membrane aperture is 1.00 μm,
20 DEG C of temperature, pressure is 0.2 MPa, and flow velocity is 20 kg/h, removes the impurity such as oil removal, bacterium and macromolecule colloid, obtains
Microfiltration membranes permeate；Ultrafiltration membrane aperture be 0.030 μm, 20 DEG C of temperature, the MPa of pressure 0.2, flow velocity be 20 kg/h, remove colloid,
Protein, microorganism and larger molecular organicses, obtain milipore filter permeate；Nanofiltration membrane aperture is 0.002 μm, 20 DEG C of temperature, pressure
The MPa of power 0.9, the kg/h of flow velocity 40, molecular cut off are 100 ~ 2000 leaflet bush cinqefoil extracted component, obtain NF membrane retention
Component；
（4）By NF membrane retention component freeze-drying, freeze-drying condition is -70 DEG C of pre-freeze 2h, and -70 DEG C are freeze-dried 24h, very
Reciprocal of duty cycle 7Pa, obtains a kind of leaflet bush cinqefoil extract D 50.64g.
2nd, the detection of leaflet bush cinqefoil extract
（1）Terpenoid content in leaflet bush cinqefoil extract：With the two of embodiment 1（1）It is identical；
（2）Flavonoid content in leaflet bush cinqefoil extract：With the two of embodiment 1（2）It is identical；
（3）Inhibitory action of the leaflet bush cinqefoil extract to human nerve cell oncocyte SH-SY5Y：With the two of embodiment 1（3）
It is identical.
After testing, terpenoid content 1.08%, flavones in the leaflet bush cinqefoil extract that the inventive method is prepared
Kind compound content 5.24%, SH-SY5Y semi-inhibit rate IC50For 105.34 μ g/mL.
Comparative example 2：The preparation method of leaflet bush cinqefoil extract and its antitumor activity application.
First, preparation method
（1）It by leaflet bush cinqefoil, dry, pulverize, obtain leaflet bush cinqefoil powder, the long 0.4cm of particle diameter；
（2）Leaflet bush cinqefoil powder 2.0kg is placed in sub-critical extraction post and extracted, extractant is water, temperature is 170 DEG C,
Extracting pressure 7MPa, extraction time is 25min, solid-liquid ratio 1:20g/mL, the number of times of extraction is 3 times；
（3）Subcritical abstraction liquid is used into microfiltration membranes, milipore filter and nanofiltration membrane treatment successively.Wherein, micro-filtration membrane aperture is 0.02 μm,
20 DEG C of temperature, pressure is 0.1 MPa, and flow velocity is 30 kg/h, removes the impurity such as oil removal, bacterium and macromolecule colloid, obtains
Microfiltration membranes permeate；Ultrafiltration membrane aperture be 0.003 μm, 20 DEG C of temperature, the MPa of pressure 0.1, flow velocity be 30 kg/h, remove colloid,
Protein, microorganism and larger molecular organicses, obtain milipore filter permeate；Nanofiltration membrane aperture is 0.001 μm, 20 DEG C of temperature, pressure
The MPa of power 0.7, the kg/h of flow velocity 50, molecular cut off are 100 ~ 2000 leaflet bush cinqefoil extracted component, obtain NF membrane retention
Component；
（4）By NF membrane retention component freeze-drying, freeze-drying condition is -70 DEG C of pre-freeze 2h, and -70 DEG C are freeze-dried 24h, very
Reciprocal of duty cycle 7Pa, obtains a kind of leaflet bush cinqefoil extract E 195.01g.
2nd, the detection of leaflet bush cinqefoil extract
（1）Terpenoid content in leaflet bush cinqefoil extract：With the two of embodiment 1（1）It is identical；
（2）Flavonoid content in leaflet bush cinqefoil extract：With the two of embodiment 1（2）It is identical
（3）Inhibitory action of the leaflet bush cinqefoil extract to human nerve cell oncocyte SH-SY5Y：With the two of embodiment 1（3）
It is identical.
After testing, terpenoid content 2.65%, flavones in the leaflet bush cinqefoil extract that the inventive method is prepared
Kind compound content 7.03%, SH-SY5Y semi-inhibit rate IC50For 80.52 μ g/mL.
In summary, the inventive method not only can effectively prepare a kind of leaflet bush cinqefoil extract, and preparation technology
Very efficiently, processing step mechanization degree is high, and preparation method mature and reliable, the especially invention are made leaflet bush cinqefoil and extracted
Thing has the activity for significantly suppressing human nerve cell oncocyte SH-SY5Y cells propagation, is a kind of the anti-of great application prospect
Tumour agent.
Claims (10)
1. the application of a kind of leaflet bush cinqefoil extract and preparation method and its antitumor activity, it is characterised in that including following step
Suddenly：
（1）By leaflet bush cinqefoil, dry, shred, particle diameter 0.4 ~ 1.0cm of length obtains leaflet bush cinqefoil powder；
（2）Leaflet bush cinqefoil powder is placed in sub-critical extraction post, ultra-pure water is conveyed and preheated into preheater by compression pump, is reached
After design temperature, it is pumped into extraction column, system pressure is reached after setting value, extraction column starts timing when filling water, extraction is certain
Extract solution flow to collection kettle through cooler and collected after time, obtains leaflet bush cinqefoil subcritical abstraction liquid；
（3）Subcritical abstraction liquid is used into microfiltration membranes, milipore filter and nanofiltration membrane treatment successively, microfiltration membranes permeate, ultrafiltration are obtained successively
Film permeate and NF membrane retention component；
（4）By NF membrane retention component freeze-drying, a kind of leaflet bush cinqefoil extract is obtained.
2. according to the method described in claim 1, it is characterised in that：Step（2）In, the subcritical abstraction agent is water, temperature
For 140 DEG C ~ 160 DEG C, 4 ~ 6MPa of extracting pressure, extraction time is 10min ~ 40min, solid-liquid ratio 1:10g/mL ~1:30g/mL,
The number of times of extraction is 2 ~ 4 times.
3. according to the method described in claim 1, it is characterised in that：Step（3）In, the micro-filtration membrane aperture that micro-filtration film process is used
For 0.02 μm ~ 8.00 μm, 20 DEG C of the temperature of processing, pressure is 0.1 ~ 0.3MPa, and flow velocity is 10 ~ 30kg/h, obtains microfiltration membranes saturating
Cross liquid.
4. according to the method described in claim 1, it is characterised in that：Step（3）In, the ultrafiltration membrane aperture that ultrafiltration membrane treatment is used
For 0.003 μm ~ 0.090 μm, 20 DEG C of temperature, 0.1 ~ 0.3MPa of pressure, flow velocity is 10 ~ 30 kg/h, obtains milipore filter transmission
Liquid.
5. according to the method described in claim 1, it is characterised in that：Step（3）In, the nanofiltration membrane aperture that nanofiltration membrane treatment is used
For 0.001 μm ~ 0.002 μm, 20 DEG C of temperature, 0.7 ~ 0.9MPa of pressure, the kg/h of flow velocity 30 ~ 50 obtain NF membrane retention component.
6. according to the method described in claim 1, it is characterised in that：Step（4）In, the freeze-drying program be -60 DEG C ~ -
80 DEG C of pre-freezes 1 ~ 3h, -60 DEG C ~ -80 DEG C freeze-dryings 12 ~ 36h, 5 ~ 10Pa of vacuum.
7. according to the method described in claim 1, it is characterised in that：Step（4）In, in described leaflet bush cinqefoil extract,
Terpenoid content 5.42% ~ 17.61%.
8. according to the method described in claim 1, it is characterised in that：Step（4）In, in described leaflet bush cinqefoil extract,
Flavonoid content 7.35% ~ 30.82%.
9. according to the method described in claim 1, it is characterised in that：Step（4）In, described leaflet bush cinqefoil extract, tool
There is the propagation for significantly suppressing human neuroblastoma cells SH-SY5Y.
10. the leaflet bush cinqefoil extract that claim 1 ~ 6 any one methods described is prepared.
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| CN115721726A (en) * | 2022-07-27 | 2023-03-03 | 江苏长泰药业有限公司 | Nasal cavity product for preventing or relieving cold symptoms and preparation method thereof |
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| CN109966191A (en) * | 2017-12-28 | 2019-07-05 | 伽蓝（集团）股份有限公司 | A kind of bush cinqefoil flower extract and its new opplication |
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