CN105693876B - Shiny-leaved yellowhorn leaf polyose extract, extracting method and its application - Google Patents
Shiny-leaved yellowhorn leaf polyose extract, extracting method and its application Download PDFInfo
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- CN105693876B CN105693876B CN201610089260.XA CN201610089260A CN105693876B CN 105693876 B CN105693876 B CN 105693876B CN 201610089260 A CN201610089260 A CN 201610089260A CN 105693876 B CN105693876 B CN 105693876B
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- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
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Abstract
The invention belongs to technical field of plant extraction, is related to a kind of shiny-leaved yellowhorn leaf polyose extract, extracting method and its application.Solve the prior art technical problem low to shiny-leaved yellowhorn leaf utilization ratio.It comprises the following steps：A, degreasing, dry shiny-leaved yellowhorn leaf is taken, addition petroleum ether and stirring is uniform, stratification, degreasing shiny-leaved yellowhorn leaf is obtained after filtering out petroleum ether；B, decolourize, add ethanol in degreasing shiny-leaved yellowhorn leaf, stir, decolouring shiny-leaved yellowhorn leaf is obtained after filtering out ethanol；C, ultrasonic extraction, pure water is added in decolouring shiny-leaved yellowhorn leaf, ultrasonic extraction, removal shiny-leaved yellowhorn leaf is filtered and obtains ultrasonic extraction liquid；D, it is concentrated and dried, is dried after ultrasonic extraction liquid is concentrated, obtain shiny-leaved yellowhorn leaf polyose extract.Ultrasonic wave added of the present invention can greatly improve shiny-leaved yellowhorn leaf polyose recovery rate, be a high effective way of shiny-leaved yellowhorn leaf polyose extraction, and external activity result of study shows that shiny-leaved yellowhorn leaf polyose has certain active anticancer and good Scavenging ability.
Description
Technical field
The invention belongs to technical field of plant extraction, is related to a kind of shiny-leaved yellowhorn, more particularly to a kind of shiny-leaved yellowhorn leaf polyose carries
Take thing, extracting method and its application.
Background technology
Shiny-leaved yellowhorn (scientific name：Xanthoceras sorbifolium Bunge), Sapindaceae, shiny-leaved yellowhorn category machaka
Or dungarunga, it is high up to 5 meters；Sprig maroon is sturdy, and leaf connecting handle length is up to 30 centimetres；Leaflet is to life, and both sides are slightly asymmetric, top
Hold tapering, base portion wedge shape, there is sharp keen sawtooth at edge, and the inflorescence basidixed of hermaphrodite flower, male inflorescence axillary, uprightly, total bennet are short, petal
White, base portion aubergine or yellow, the horn shape appendage of floral disc is orange-yellow, and filigree is hairless；Capsule is up to 6 centimetres；Seed black
It is and glossy.Spring blooms, autumn firstfruits.
Shiny-leaved yellowhorn originates in northwest China, is mainly distributed on the province such as Shaanxi, Shanxi, Hebei, Inner Mongol, there is higher food
With, it is medicinal, the economic value such as view and admire.There are wind-damp dispelling, swelling and pain relieving and other effects, be clinically mainly used in treating rheumatism joint
The disease such as heat, skin wind-heat, infantile enuresis in scorching, rheumatism, and it has been included in as medicinal part the Chinese Pharmacopoeia of version in 1977.Separately
Outside, shiny-leaved yellowhorn adaptability is very strong, and drought-resistant, barren, well developed root system, sprout tiller strength is strong, grows fast, solid morning, yield height, life-span
It is long, economic value is big, be precious view and admire and important woody oil tree species.Shiny-leaved yellowhorn is classified as woody fuel oil by country at present
Energy chief species, its Breeding and popularization are paid much attention to by country, and it is imperative to greatly develop shiny-leaved yellowhorn.
The existing shiny-leaved yellowhorn cultivated area in the whole nation develops based on being extracted oil with benevolence to it up to 5.35 ten thousand hm2 at present.Wen Guan
Fruit leaf contain a variety of chemical compositions (Ma Yangmin, king wear, research [J] Chinese patent drugs of shiny-leaved yellowhorn study on chemical compositions of leaves, 2010,32
(10), 1750-1753) and there is anti-inflammatory analgetic, hemostasia and detumescence.At present, the utilization to shiny-leaved yellowhorn leaf are with tealeaves
Based on, as Chinese patent literature discloses a kind of shiny-leaved yellowhorn leaf tea and its applies [application number：201310217440.8], it is a kind of with
Shiny-leaved yellowhorn leaf is primary raw material, and the drink made using black tea processing technology supports tea [application number：201110238087.2], Yi Zhonghan
There is the tealeaves [patent No. of bunkankasaponin bioactive substance:200710018943.7].
The content of the invention
A kind of regarding the issue above, the present invention provides extraction method of polysaccharides of the high shiny-leaved yellowhorn leaf of recovery rate.
It is a further object of the present invention to provide a kind of polyoses extract of shiny-leaved yellowhorn leaf.
Another object of the present invention is to provide a kind of application of shiny-leaved yellowhorn leaf polyose extract in cancer cell is suppressed.
The further object of the present invention is to provide a kind of application of shiny-leaved yellowhorn leaf polyose extract in radicals scavenging.
To reach above-mentioned purpose, present invention employs following technical proposal：A kind of extraction method of polysaccharides of shiny-leaved yellowhorn leaf, bag
Include following steps：
A, degreasing, dry shiny-leaved yellowhorn leaf is taken, addition petroleum ether and stirring is uniform, stratification, is taken off after filtering out petroleum ether
Fat shiny-leaved yellowhorn leaf；
B, decolourize, add ethanol in degreasing shiny-leaved yellowhorn leaf, stir, decolouring shiny-leaved yellowhorn leaf is obtained after filtering out ethanol；
C, ultrasonic extraction, pure water is added in decolouring shiny-leaved yellowhorn leaf, ultrasonic extraction, removal shiny-leaved yellowhorn leaf is filtered and is surpassed
Sound extract solution；
D, it is concentrated and dried, is dried after ultrasonic extraction liquid is concentrated, obtain shiny-leaved yellowhorn leaf polyose extract.
In the extraction method of polysaccharides of above-mentioned shiny-leaved yellowhorn leaf, in step D, added after the concentration of ultrasonic extraction liquid liquid appropriate
Washes of absolute alcohol, filtered after being sufficiently stirred and obtain filter residue, shiny-leaved yellowhorn leaf polyose extract is obtained after filter residue is freeze-dried.
In the extraction method of polysaccharides of above-mentioned shiny-leaved yellowhorn leaf, in step D, ultrasonic extraction liquid is concentrated into five points of stoste
One of obtain concentrate, add the absolute ethyl alcohol of 4 times of volumes of concentrate, filtered after being sufficiently stirred and obtain filter residue, filter residue freezing
It is dried to obtain shiny-leaved yellowhorn leaf polyose extract.
In the extraction method of polysaccharides of above-mentioned shiny-leaved yellowhorn leaf, in step, the fresh text of described drying shiny-leaved yellowhorn leaf
Hat fruit leaf is crushed after drying in the shade indoors and is made, and in stepb, described ethanol is 95% ethanol or absolute ethyl alcohol, in step C
In, solid-liquid ratio is to dry shiny-leaved yellowhorn leaf：Pure water=1:10-1:30, the ultrasonic extraction time is 10-60min, and ultrasonic power is
20-100W, ultrasonic extraction temperature are 25-90 DEG C.
In the extraction method of polysaccharides of above-mentioned shiny-leaved yellowhorn leaf, in step, described drying shiny-leaved yellowhorn leaf is crushed to
40-60 mesh, dry shiny-leaved yellowhorn leaf：Petroleum ether=1:1-1:30, in stepb, solid-liquid ratio is to dry shiny-leaved yellowhorn leaf：95% ethanol
=1:1-1:30, in step C, solid-liquid ratio is to dry shiny-leaved yellowhorn leaf：Pure water=1:15, the ultrasonic extraction time is 40min, is surpassed
Acoustical power is 80W, and ultrasonic extraction temperature is 80 DEG C.
According to shiny-leaved yellowhorn leaf polyose extract made from above-mentioned extracting method.
Application of the shiny-leaved yellowhorn leaf polyose extract in cancer cell is suppressed.
In the applications described above, described cancer cell is liver cancer cells.
In the applications described above, described liver cancer cells behaviour hepatocellular carcinoma H22.
Application of the shiny-leaved yellowhorn leaf polyose extract in radicals scavenging.
Compared with prior art, the advantage of the invention is that：Ultrasonic wave added can greatly improve carrying for shiny-leaved yellowhorn leaf polyose
Efficiency is taken, is a high effective way of shiny-leaved yellowhorn leaf polyose extraction.External activity result of study shows that shiny-leaved yellowhorn leaf polyose has
Certain active anticancer and good Scavenging ability, there are potentiality of the exploitation for Related products such as medicine and cosmetics,
This provides new thinking for the comprehensive exploitation of shiny-leaved yellowhorn resource with utilizing.
Brief description of the drawings
Fig. 1 is suppression figure of the polyoses extract of the offer of embodiment 3 to HepG2 cells；
Fig. 2 is the polyoses extract and the curve map of the DPPH clearance rates of VC reference substances that embodiment 3 provides.
Embodiment
Embodiment 1
Fresh shiny-leaved yellowhorn leaf 1000g is taken, dried in the shade under normal temperature laboratory, be crushed to standby after 60 mesh, take that 100g is dry, crushes
Drying shiny-leaved yellowhorn leaf afterwards adds 100g petroleum ethers, stirs, filtered after stratification, filter out oil in flask
Ether, degreasing shiny-leaved yellowhorn leaf is obtained, the ethanol of 100g 95% is added in degreasing shiny-leaved yellowhorn leaf, is sufficiently stirred, is filtered after standing, filtered
Except ethanol, decolouring shiny-leaved yellowhorn leaf is obtained, 1000g pure water is added in decolouring shiny-leaved yellowhorn leaf, is placed in ultrasonic sink ultrasonic
Extraction, ultrasonic power 20W, extraction time 10min, ultrasonic temperature is 25 DEG C, after the completion of extraction, filters, removes shiny-leaved yellowhorn
Leaf, ultrasonic extraction liquid is obtained, ultrasonic extraction liquid is freeze-dried, obtain shiny-leaved yellowhorn leaf polyose extract.
It will be appreciated by those skilled in the art that the pure water of extraction is equal to deionized water, process water etc..
Embodiment 2
Fresh shiny-leaved yellowhorn leaf 1000g is taken, dried in the shade under normal temperature laboratory, be crushed to standby after 40 mesh, take that 100g is dry, crushes
Drying shiny-leaved yellowhorn leaf afterwards adds 3000g petroleum ethers, stirs, filtered after stratification, filter out oil in flask
Ether, degreasing shiny-leaved yellowhorn leaf is obtained, the ethanol of 3000g 95% is added in degreasing shiny-leaved yellowhorn leaf, is sufficiently stirred, is filtered after standing, filtered
Except ethanol, decolouring shiny-leaved yellowhorn leaf is obtained, 3000g pure water is added in decolouring shiny-leaved yellowhorn leaf, is placed in ultrasonic sink ultrasonic
Extraction, ultrasonic power 100W, extraction time 60min, ultrasonic temperature is 90 DEG C, after the completion of extraction, filters, removes shiny-leaved yellowhorn
Leaf, ultrasonic extraction liquid is obtained, ultrasonic extraction liquid is concentrated into 1/5th of original volume, the ultrasonic extraction liquid four added after concentration
The absolute ethyl alcohol of times volume, forms suspension after being sufficiently stirred, removes ethanol water after suspension is filtered, obtain filter residue,
Filter residue is freeze-dried, and obtains shiny-leaved yellowhorn leaf polyose extract, and it is 12.42% that recovery rate is shown after weighing.
Embodiment 3
Fresh shiny-leaved yellowhorn leaf 1000g is taken, dried in the shade under normal temperature laboratory, be crushed to standby after 50 mesh, take that 100g is dry, crushes
Drying shiny-leaved yellowhorn leaf afterwards adds 2000g petroleum ethers, stirs, filtered after stratification, filter out oil in flask
Ether, degreasing shiny-leaved yellowhorn leaf is obtained, 2000g absolute ethyl alcohols are added in degreasing shiny-leaved yellowhorn leaf, is sufficiently stirred, is filtered after standing, filtered
Except ethanol, decolouring shiny-leaved yellowhorn leaf is obtained, 1500g pure water is added in decolouring shiny-leaved yellowhorn leaf, is placed in ultrasonic sink ultrasonic
Extraction, ultrasonic power 80W, extraction time 40min, ultrasonic temperature is 80 DEG C, after the completion of extraction, filters, removes shiny-leaved yellowhorn
Leaf, ultrasonic extraction liquid is obtained, ultrasonic extraction liquid is concentrated into a quarter of original volume, the ultrasonic extraction liquid three added after concentration
The absolute ethyl alcohol of times volume, forms suspension after being sufficiently stirred, removes ethanol water after suspension is filtered, obtain filter residue,
Filter residue is freeze-dried, and obtains shiny-leaved yellowhorn leaf polyose extract, and it is 17.90% that recovery rate is shown after weighing.
Embodiment 4-9
The present embodiment is substantially the same manner as Example 3, and difference is as shown in the table,
As seen from the above table, the yield of shiny-leaved yellowhorn leaf polyose extract changes bigger with extraction time, and extraction time is more than
After 40min, yield is not further added by substantially, therefore it is the optimum extraction time to select 40min.
Embodiment 10-14
The present embodiment is substantially the same manner as Example 3, and difference is as shown in the table,
As seen from the above table, when the solid-liquid ratio of extraction is more than 1: 15, yield still increases as solid-liquid ratio increases, still
Increase trend slows down, and solid-liquid ratio is bigger, and subsequent concentration is difficult, therefore selects 1: the 15 optimal solid-liquid ratio for extraction.
Embodiment 15-21
The present embodiment is substantially the same manner as Example 3, and difference is as shown in the table,
As seen from the above table, rise with ultrasonic temperature, yield rises, and yield rises larger especially between 70-80 DEG C, works as temperature
Yield is not further added by substantially after degree is higher than 80 DEG C, is tended towards stability, so 80 DEG C of selection is optimum extraction temperature.
Embodiment 22-27
The present embodiment is substantially the same manner as Example 3, and difference is as shown in the table,
As seen from the above table, with the increase of ultrasonic power, recovery rate increase, must increase after ultrasonic power is more than 80W
In stable state, it is thus determined that 80W is optimum extraction frequency.
Comparative example 1
This comparative example is substantially the same manner as Example 3, and difference exists, ultrasonic power 0W, namely does not use ultrasonic extraction,
Recovery rate is 13.30%.Understand, the yield of ultrasonic extraction is higher.
Application examples 1
1) culture of HepG2 cells
HepG2 cells are placed in 25mL plastic culture bottles, complete DMEM in high glucose culture medium is added and (contains 10% tire ox blood
Clearly, 100U/L penicillin and 100mg/L streptomysins), it is placed in 37 DEG C of constant incubators containing 5%CO2.Treat cell growth extremely
90% when converging, and with 0.25% pancreatin had digestive transfer culture, 3d is passed on 1 time.
2) mtt assay detection comparative survival rate of cells
HepG2 cells are inoculated in 96 orifice plates with 1 × 104/density per hole, and culture 12h makes cell fully adherent.Abandon
Culture medium is removed, is separately added into the shiny-leaved yellowhorn leaf water extraction positions of various concentrations, 30% ethanol water extract part, the extraction of 50% ethanol water
Position, 70% ethanol water extract part and 95% ethanol water extract part.The 4h before cell culture terminates, add 15 μ L's per hole
MTT.After culture terminates, culture medium is sucked, 150 μ L DMSO is added per hole, 37 DEG C of concussion 10min, is examined with multi-function microplate reader
The absorbance surveyed at 450nm.
As shown in figure 1, shiny-leaved yellowhorn leaf polyose has the activity for suppressing human hepatoma cell HepG 2 proliferation, it is between concentration
Good dose-effect relationship, when sample concentration is 2.5mg/mL, cell proliferation inhibition rate 74.27%.
Application examples 2
1) the preparation precision of DPPH standard liquids weighs DPPH standard items 12mg, is dissolved and is settled to methanol
250ml, keep in dark place at low temperature, it is standby.Before use will with methanol dilution 2 times measure its absorbance.
2) preparation of the solution of Vc and sample measures ascorbic acid and sample 25mg respectively, is dissolved and is settled to methanol
25ml volumetric flasks.It is appropriate to measure above-mentioned solution for precision respectively, is successively 0.01mg/ml with methanol dilution to mass concentration,
The sample solution of six gradients of 0.04mg/ml, 0.08mg/ml, 0.12mg/ml, 0.16mg/ml, 0.20mg/ml.
3) test method：Take 2ml DPPH titer 2ml methanol solutions to measure to mix, reacting at normal temperature without light 15min,
Absorbance (A blank) is measured at 517nm, its absorbance (A samples) of methanol measurement, positive control are replaced with each concentration samples solution
Reference sample method.
DPPH clearance rates=(A blank-A samples)/A blank × 100%
As illustrated, shiny-leaved yellowhorn leaf polyose has stronger scavenging capacity to DPPH free radicals, and there is dose-effect relationship,
To DPPH free radical scavenging activities just up to 90.78% when concentration is 0.2mg/ml.
Specific embodiment described herein is only to spirit explanation for example of the invention.Technology belonging to the present invention is led
The technical staff in domain can be made various modifications or supplement to described specific embodiment or be replaced using similar mode
Generation, but without departing from the spiritual of the present invention or surmount scope defined in appended claims.
Claims (6)
1. a kind of extraction method of polysaccharides of shiny-leaved yellowhorn leaf, it is characterised in that comprise the following steps：
A, degreasing, dry shiny-leaved yellowhorn leaf is taken, addition petroleum ether and stirring is uniform, stratification, and degreasing text is obtained after filtering out petroleum ether
It is preced with fruit leaf；
B, decolourize, add ethanol in degreasing shiny-leaved yellowhorn leaf, stir, decolouring shiny-leaved yellowhorn leaf is obtained after filtering out ethanol；
C, ultrasonic extraction, adds pure water, ultrasonic extraction in decolouring shiny-leaved yellowhorn leaf, and suction filtration removal shiny-leaved yellowhorn leaf obtains ultrasound and carried
Take liquid；
D, it is concentrated and dried, is dried after ultrasonic extraction liquid is concentrated, obtain shiny-leaved yellowhorn leaf polyose extract.
2. the extraction method of polysaccharides of shiny-leaved yellowhorn leaf according to claim 1, it is characterised in that in step D, ultrasonic extraction
Appropriate washes of absolute alcohol is added after the concentration of liquid liquid, is filtered after being sufficiently stirred and obtains filter residue, text is obtained after filter residue is freeze-dried
It is preced with fruit leaf polyose extract.
3. the extraction method of polysaccharides of shiny-leaved yellowhorn leaf according to claim 2, it is characterised in that in step D, ultrasonic extraction
What liquid was concentrated into stoste 1/5th obtains concentrate, adds the absolute ethyl alcohol of 4 times of volumes of concentrate, is filtered after being sufficiently stirred
Filter residue is obtained, filter residue is freeze-dried to obtain shiny-leaved yellowhorn leaf polyose extract.
4. the extraction method of polysaccharides of shiny-leaved yellowhorn leaf according to claim 1, it is characterised in that in step, described is dry
Dry shiny-leaved yellowhorn leaf dried in the shade indoors with fresh shiny-leaved yellowhorn leaf after crush be made, in stepb, described ethanol be 95% ethanol or
Absolute ethyl alcohol, in step C, solid-liquid ratio is to dry shiny-leaved yellowhorn leaf：Pure water=1:10-1:30, the ultrasonic extraction time is 10-
60min, ultrasonic power 20-100W, ultrasonic extraction temperature are 25-90 DEG C.
5. the extraction method of polysaccharides of shiny-leaved yellowhorn leaf according to claim 4, it is characterised in that in step, described is dry
Dry shiny-leaved yellowhorn leaf is crushed to 40-60 mesh, dries shiny-leaved yellowhorn leaf：Petroleum ether=1:1-1:30, in stepb, solid-liquid ratio is drying
Shiny-leaved yellowhorn leaf：95% ethanol=1:1-1:30, in step C, solid-liquid ratio is to dry shiny-leaved yellowhorn leaf：Pure water=1:15, ultrasound
Extraction time is 40min, and ultrasonic power 80W, ultrasonic extraction temperature is 80 DEG C.
6. shiny-leaved yellowhorn leaf polyose extract made from the extracting method according to claim 1-5 any one.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109232757A (en) * | 2018-08-31 | 2019-01-18 | 吉首大学 | Walnut Leaves polyoses extract and application |
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| CN106753912A (en) * | 2016-11-22 | 2017-05-31 | 陕西中医药大学 | A kind of shinyleaf yellowhorn oil handmade soap and preparation method thereof |
| CN111171111B (en) * | 2019-12-31 | 2021-02-26 | 北京海达华环境科技有限公司 | Method for continuously extracting crude protein, polysaccharide and cellulose from fresh xanthoceras sorbifolia shells |
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| CN101242850A (en) * | 2003-10-09 | 2008-08-13 | 太平洋艾瑞有限公司 | Composition, function and use of xanthoceras sorbifolia extract and compound isolated from same, method for preparing same |
| CN101948897A (en) * | 2010-09-11 | 2011-01-19 | 山西金绿禾燕麦研究所 | Method for extracting shinyleaf yellowhorn oil, shinyleaf yellowhorn polysaccharide and shinyleaf yellowhorn protolysate peptide from shinyleaf yellowhorn |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109232757A (en) * | 2018-08-31 | 2019-01-18 | 吉首大学 | Walnut Leaves polyoses extract and application |
| CN109232757B (en) * | 2018-08-31 | 2020-12-01 | 吉首大学 | Walnut leaf polysaccharide extract and application thereof |
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