CN104857036B - A kind of extracting method and product of euscaphis konishii active material - Google Patents
A kind of extracting method and product of euscaphis konishii active material Download PDFInfo
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- CN104857036B CN104857036B CN201410430542.2A CN201410430542A CN104857036B CN 104857036 B CN104857036 B CN 104857036B CN 201410430542 A CN201410430542 A CN 201410430542A CN 104857036 B CN104857036 B CN 104857036B
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Abstract
Increase the present invention relates to a kind of extracting method of euscaphis konishii active material, product and cancer inhibitor cell and apply；Main process steps：Euscaphis konishii material after dries pulverizing and ethanol are loaded into extraction vessel by solid-liquid ratio 1: 10, evaporative condenser reflux is installed, flowed back under 80 DEG C of heating water baths, 2 hours every time, totally 3 times, filtering, merge 3 filtrates and collect rotary evaporation in vacuo, ethanol is reclaimed, final 50 DEG C of drying obtain crude extract S；S is dissolved with isometric distilled water in ultrasonic vibration, extracted 3 times by sample solution and ethyl acetate 1: 3, merge upper strata ethyl acetate phase liquid and collect rotary evaporation in vacuo, recovery ethyl acetate obtains ethyl acetate phase extract, extract S1 is obtained in 50 DEG C of drying, method as provided by the present invention and product.Extraction separation method is simple, aboundresources, and extract reaches 81.63% to liver cancer cells inhibiting rate, close with 5 fluorouracils (90.81%).
Description
Technical field
Patent of the present invention belongs to extracting method, product and the application field of active substance of plant, more particularly to a kind of knuckle-tooth
The extracting method and product of wild crow Chinese toon active material.
Background technology
Euscaphis konishii（ Euscaphis konishii）It is Staphyleaceae open country crow Toona（Euscaphis）Plant, alias
Chicken gizzard, corn eyeball, dog head green pepper.As a kind of China's traditional Chinese herbal medicine among the people, there is good pharmaceutical ground and long
History, primary treatment stomachache, traumatic injury, rheumatalgia pain, dizziness, rush down disease, dysentery.According to relevant report, from wild crow Chinese toon blade
Isolated three kinds of ester type compounds in extract, it is found that Euscapholide class compound can suppress HeLa undesired cell proliferations,
Promote apoptosis of tumor cells, there is important practical application meaning, but wild crow Chinese toon without the artificial breeding carried out in scale with
Cultivation, resource lack；Euscaphis konishii belongs to together with wild crow Chinese toon, and the seminar where inventor has breached artificial breeding technique,
And large-scale artificial breeding is carried out；But extraction and application field about euscaphis konishii active material also nobody are carried out
Research, also without the report of correlation.
The content of the invention
China coast province, city are the high Burst Regions of liver cancer, and tremendous influence, the object of the invention are produced to people's health
It is to be directed to this provincial characteristics, there is provided one kind separation screening out of euscaphis konishii plant, which has, suppresses the pernicious increasing of liver cancer cells
Method, product and the application for suppressing liver cancer cells malignant proliferation for the compound grown.
The purpose of the present invention is realized as follows：
A kind of extracting method of euscaphis konishii active material, its key step
A. euscaphis konishii plant, chopping, 50 DEG C of drying, crushing are taken；
B. A powders are taken, by feed liquid weight than 1：10 95% ethanol of addition, mixing are fitted into round-bottomed flask, install evaporative condenser
Reflux, flowed back 2 hours under 80 DEG C of heating water baths, filter to get filtrate a；
C. B filter residues are taken, repeat B extraction steps, filter to get filtrate b；
D. C filter residues are taken, repeat C extraction steps, filter to get filtrate c；
E. merging filtrate a, b, c, evaporated on rotary evaporation in vacuo device, reclaim ethanol, final 50 DEG C of drying obtain thick
Product extract S；
F. extract S measurements are weighed, with etc. weight distilled water ultrasonic vibration dissolve, by 1：3 add ethyl acetate
After extraction 3 times, merge upper strata ethyl acetate phase extract and be evaporated under reduced pressure to obtain ethyl acetate phase extract, obtained in 50 DEG C of drying
Extract S1,4 DEG C save backup.
A kind of extracting method of above-mentioned euscaphis konishii active material, the euscaphis konishii plant used in it include each
Kind organ, it is one or more mixing of branch, leaf, root, flower, kind skin, seed.
A kind of euscaphis konishii active material product, it is the extract S1 obtained using the above method.
A kind of euscaphis konishii active material suppresses liver cancer cells malignant proliferation, is the extraction obtained using the above method
Thing S1, for suppressing liver cancer cells malignant proliferation.
It is simple using said extracted separation method, it is practical, and raw material euscaphis konishii can a large amount of artificial cultures, money
Source is enriched, and extract is obvious to liver cancer cells inhibitory action, can reach 81.63%, positive with largely being used in doctor's medical treatment at present
According to medicine 5 FU 5 fluorouracil（90.81%）It is fairly close, Small side effects.
Embodiment 1
Euscaphis konishii branches and leaves material after dries pulverizing and 95% ethanol are pressed into solid-liquid ratio 1：10 mixing load round-bottomed flask
In, evaporative condenser reflux is installed, flowed back under 80 DEG C of heating water baths, 2 hours every time, totally 3 times, filtering, merges 3 filtrates
And rotary evaporation in vacuo is collected, ethanol is reclaimed, final 50 DEG C of drying obtain crude extract S；Obtained medicinal extract survey will be dried again
Amount weigh after with etc. weight distilled water ultrasonic vibration dissolve, by sample solution and ethyl acetate 1：3 carry out liquid-liquid extraction 3
It is secondary, merge upper strata ethyl acetate phase, merge 3 filtrates and collect rotary evaporation in vacuo, recovery ethyl acetate obtains ethyl acetate
Phase extract, extract S1 is obtained in 50 DEG C of drying, weighs and measures sample yield, 4 DEG C save backup；
BEL- of several effectors in vitro culture is determined using Methyl Thiazolylte trazolium (MTT) method
The inhibitory action of 7404 liver cancer cells.Exponential phase cell is washed twice with PBS solution, adds 0.25% trypsase 37
Degree Celsius, digest 5 minutes, make its de- wall, be resuspended with containing 10% or so calf serum RPMI1640 nutrient solutions into unicellular outstanding
Liquid, counted with blood cell counting plate, be inoculated into every 104 cells in hole in 96 orifice plates, the calf serum per 200 microlitres of Kong Zhongjia
RPMI1640 nutrient solutions, 96 orifice plates are put into CO2Incubator 24 hours, then former culture medium is suctioned out and abandons it, addition contains medicine
100 microlitres and 100 microlitres nutrient solutions of solution continue culture 48 hours, to the time after add 20 microlitres of 5mg/mlMTT solution, culture
Terminate culture within 4 hours, add DMSO150 microlitres and shake 10 minutes, enzyme micro-plate reader measure is per hole light absorbs under 550nm wavelength
Value（A values）, finally calculate inhibiting rate using formula：
Inhibiting rate（%）=（Control wells A values-experimental port A values）/ control wells A value × 100% calculates final inhibiting rate,
As a result such as table 1.
Inhibitory action of the S1 of table 1 to BEL-7404 liver cancer cells
* positive control medicine selection 5 FU 5 fluorouracil
Embodiment 2
Such as the method for embodiment 1, using the bar of euscaphis konishii as material, extract S1 is obtained, its 1000 mg/L is to BEL-
The inhibition of 7404 liver cancer cells is higher than 81.63%.
Embodiment 3
Such as the method for embodiment 1, using the root of euscaphis konishii as material, extract S1 is obtained, its 1000 mg/L is to BEL-
The inhibition of 7404 liver cancer cells is higher than 81.63%.
Embodiment 4
Such as the method for embodiment 1, using the seed of euscaphis konishii as material, extract S1 is obtained, its 1000 mg/L is to BEL-
The inhibition of 7404 liver cancer cells is higher than 81.63%.
Embodiment 5
Such as the method for embodiment 1, using the kind skin of euscaphis konishii as material, extract S1 is obtained, its 1000 mg/L is to BEL-
The inhibition of 7404 liver cancer cells is higher than 81.63%.
Embodiment 6
Such as the method for embodiment 1, using the branches and leaves in euscaphis konishii winter as material, the branches and leaves include and its accompanying kind
Skin, seed, obtain extract S1, and its 1000 mg/L is higher than 81.63% to the inhibition of BEL-7404 liver cancer cells.
Claims (4)
1. a kind of extracting method of euscaphis konishii active material, it is characterized in that
A. euscaphis konishii plant, chopping, 50 DEG C of drying, crushing are taken；
B. A powders are taken, by feed liquid weight than 1：10 95% ethanol of addition, mixing are fitted into round-bottomed flask, and installation evaporative condenser returns
Device is flowed, is flowed back 2 hours under 80 DEG C of heating water baths, filter to get filtrate a；
C. B filter residues are taken, repeat B extraction steps, filter to get filtrate b；
D. C filter residues are taken, repeat C extraction steps, filter to get filtrate c；
E. merging filtrate a, b, c, evaporated on rotary evaporation in vacuo device, reclaim ethanol, final 50 DEG C of drying obtain crude product leaching
Cream S；
F. medicinal extract S measurements are weighed, with etc. weight distilled water ultrasonic vibration dissolve, by 1：3 add ethyl acetate extraction 3
After secondary, merge upper strata ethyl acetate phase extract and be evaporated under reduced pressure to obtain ethyl acetate phase extract, medicinal extract is obtained in 50 DEG C of drying
S1,4 DEG C save backup.
A kind of 2. extracting method of euscaphis konishii active material according to claim 1, it is characterized in that knuckle-tooth used is wild
Crow Chinese toon plant includes various organs, is one or more mixing of branch, leaf, root, flower, kind skin, seed.
A kind of 3. euscaphis konishii active material product, it is characterized in that the medicinal extract S1 products obtained using claim 1 method.
4. a kind of euscaphis konishii active material is preparing the application in suppressing liver cancer cells malignant proliferation medicine, it is characterized in that institute
The euscaphis konishii active material stated is the medicinal extract S1 described in claim 1.
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| CN104857036A CN104857036A (en) | 2015-08-26 |
| CN104857036B true CN104857036B (en) | 2018-01-12 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106727767A (en) * | 2017-01-22 | 2017-05-31 | 福建农林大学 | Application of the euscaphis konishii extract in medicines resistant to liver cancer is prepared |
| CN110845632B (en) * | 2019-11-03 | 2023-02-10 | 福建农林大学 | Novel preparation method of euscaphis konishii hayata pericarp polysaccharide and product application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101595846A (en) * | 2009-07-21 | 2009-12-09 | 福建农林大学 | Euscaphis konishii blade high frequency regenerating system is set up and method for quickly breeding |
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| KR100913917B1 (en) * | 2008-08-27 | 2009-08-26 | 성균관대학교산학협력단 | Plant Extracts Modulating Neurotransmitter Release |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101595846A (en) * | 2009-07-21 | 2009-12-09 | 福建农林大学 | Euscaphis konishii blade high frequency regenerating system is set up and method for quickly breeding |
Non-Patent Citations (2)
| Title |
|---|
| "福建野鸦椿籽中的酚酸类化学成分";黄云等;《中草药》;20140930;第45卷(第18期);第2611-2613页 * |
| "野鸦椿对Hela细胞的抗增殖作用及其机制的初步研究";左敏等;《癌变.畸变.突变》;20080930;第20卷(第5期);第350-353页，尤其是第351页左栏第1段和最后1段，第352页左栏第3段 * |
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Effective date of registration: 20171117 Address after: 350002 Fujian Agriculture And Forestry University, Fuzhou, Fujian Applicant after: Fujian Agricultural and Forestry University Applicant after: Scientific and Technical Developnent General Co., Fujian Agriculture Univ. Address before: 350002 Cangshan District, Fuzhou, Fujian Province, Fujian Agriculture And Forestry University Applicant before: Scientific and Technical Developnent General Co., Fujian Agriculture Univ. |
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Effective date of registration: 20200715 Address after: Area a, No. 199, No. 2 workshop, hongku2 District, hongku village, Hanjiang Town, Shishi City, Quanzhou City, Fujian Province Patentee after: Fujian Yaoyi Pharmaceutical Co., Ltd Address before: 350002 Fujian Agriculture And Forestry University, Jinshan, Fujian, Fuzhou Co-patentee before: FUJIAN NONGDA TECHNOLOGY DEVELOPMENT Corp. Patentee before: FUJIAN AGRICULTURE AND FORESTRY University |
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