CN105218695B - A kind of lycium ruthenicum polysaccharide extract and preparation method thereof - Google Patents
A kind of lycium ruthenicum polysaccharide extract and preparation method thereof Download PDFInfo
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Abstract
The present invention relates to a kind of lycium ruthenicum polysaccharide extract and preparation method thereof, by clearing up, the lycium ruthenicum polysaccharide that dry, broken, removal of impurities, extraction, decolouring, removing protein, concentration, the preparation method such as purifying obtain there is the strong kidney of enriching yin, improving eyesight nourishing liver, lipid-loweringing, anti-cancer, anti-aging and other effects, while the present invention lycium ruthenicum polysaccharide preparation method technique it is simple it is easily operated, yield is higher, quality is good, is advantageously implemented industrialized production and processing.
Description
Technical field
It is specially a kind of there is the strong kidney of enriching yin, improving eyesight nourishing liver, lipid-loweringing, anti-cancer, anti-the present invention relates to plant extracts field
Lycium ruthenicum polysaccharide extract of the functions such as aging and preparation method thereof.
Background technology
Black fruit fructus lycii is Solanaceae Lycium black fruit fructus lycii (Lycium Ruthenicum Murr) dry fruit, in China
All there is fragmentary distribution on the ground such as Jiuquan, East of Qinghai Province, Xinjiang middle part, West Inner Mongolia, Ningxia and Tibet.It is grown in the mankind
The barren hill that can not be survived and riverbed sandy beach, possess extremely strong vitality, and the wild black fruit fructus lycii of Golmud of Qinghai is praised
For grassland " soft gold ".But with exploitation of the mankind to the Nature, the living space of black fruit fructus lycii is less and less, creates black fruit
This rare precious ecosystem top grade excellent tonic product of matrimony vine.Its is sweet, mild-natured, have strong kidney, nourishing liver, improving eyesight, stomach invigorating, cerebrum tonifying,
Function that is anti-ageing and stimulating the menstrual flow.Uygur medicine treats the disease such as dizziness, blurring of vision, kidney deficiency sun ruffian, in poor health with it.Tibetan medicine uses
In treatment cardiopyretic disease, heart disease, reduce cholesterol, excited cerebral nerve, enhancing immunologic function, anti-curing cancers, anti-aging, beauty
Beauty treatment, irregular menstruation, menopause etc. and drug effect is notable, also commonly use black fruit fructus lycii fruit and root skin treatment urethral calculus, tinea scabies, gums go out
The diseases such as blood.Black fruit fructus lycii fruit is rich in protein, fat, carbohydrate, free amino acid, organic acid, mineral matter, trace element, biology
The various nutritional ingredients such as alkali, vitamin C, B1, B2, calcium, magnesium, copper, zinc, manganese, iron, lead, nickel, cadmium, cobalt, chromium, potassium, sodium.Black fruit Chinese holly
Qi with red matrimony vine it is the same can tonifying kidney and benefiting sperm, the anthocyanidin (Anthocyanosides) in black fruit fructus lycii, which has, removes free radical
Effect, also cancer cell can be allowed not spread smoothly, the cell of the more health of protection by cancer cell from being corroded whereby.Seem mammary gland
The mechanism of causing a disease of cancer is in this way, therefore taking development of the anthocyanidin for breast cancer has good inhibiting effect.Black fruit Chinese holly
Qi can also prevent and treat diabetes, and with obvious antifatigue, liver protection, antitumor, dredging vascellum, decompression and protection cardiovascular system
Effect.Particularly lycium ruthenicum polysaccharide content is high, has good health value and medical value.
Polysaccharide (polysaccharide) is also known as polysaccharide, is to be gathered by the monose of more than 20 with what sugared former times key was connected to form
Compound.It is most colourful member in composition high-molecular compound family.Polysaccharide comes from higher plant, zooblast
Film, microorganism cell membrane in natural macromolecular material, be the important composition composition of all living organisms with sustaining life
Necessary structural material.People are it to be regarded as the energy source in food first to the understanding of sugar.Research to polysaccharide, most
Be early in the 1940s, but its cause as broad immune accelerator the very big attention of people to be in the sixties, process
The continuous development in more than 40 years, to polysaccharide, this kind of important living matter generates new understanding to people, this subject is turned into mesh
One of most active field is studied in preceding life science.It is found that carbohydrate functions not only as energy matter or knot in organism
Structure material, it is often more important that it take part in the various activities of cell in life science, have diversified biological function.Therefore
The research of sugar is progressively active, wherein, number molecular weight has grinding for the very active polysaccharide of strong biological activity more than thousands of
Study carefully and receive increasing attention.Physiologically active, chemical constitution and the structure-activity relationship of these active polysaccharides turn into the forward position of polysaccharide researches
Position, and make great progress.Scientific worker gradually has found that polysaccharide compound has antitumor, anti-aging, anti-sense
The bioactivity of dye, reducing blood sugar and blood lipid and treatment AIDS etc..In recent years, find the sugar chain of polysaccharide in control cell again
Division and differentiation, decisive role is played in terms of the growth and aging that adjust cell, polysaccharide has gradually shown more and more extensive
Application prospect.
Lycium ruthenicum polysaccharide is the principle active component of black fruit fructus lycii, and pharmacology is very noticeable with clinical research.Black fruit
LBP-X has extensive pharmacotoxicological effect, can promote the effect such as hematopoiesis, reducing blood lipid, liver protection and anticancer improving eyesight nourishing liver, lipid-loweringing,
Anti-cancer and anti-ageing function of waiting for a long time.
Currently available technology, which prepares the conventional extracting method of polysaccharide, to be had：Hot water extraction, acidleach formulation, alkali method and enzyme
Method.These methods all there is it is many problem of, for example destroy polysaccharide structures, Polyose extraction efficiency is low, extraction time mistake
Long and extraction process complexity etc..
The content of the invention
It is an object of the invention to provide the lycium ruthenicum polysaccharide that a kind of loss of effective components is few, quality is high, extraction cost is low
Extracting method.The lycium ruthenicum polysaccharide that this method extracts possesses with the strong kidney of enriching yin, improving eyesight nourishing liver, lipid-loweringing, anti-cancer and resisted
The functions such as aging.
Specifically, the extracting method of the present invention is as follows：
A kind of extracting method of lycium ruthenicum polysaccharide, it is characterised in that using black fruit fructus lycii, after crushing, using backflow
Method is extracted.
A kind of extracting method of lycium ruthenicum polysaccharide, it is characterised in that comprise the following steps：
1st, clear up：Black fruit fructus lycii is screened out into silt, cleaned with clear water；
2nd, dry：By the black fruit fructus lycii cleaned up at 5~30 DEG C 10~24h of low temperature drying；
3rd, crush：The black fruit fructus lycii that step 2 is obtained carries out ultramicro grinding, and it is 1~75 μm of powder to obtain particle size range
End；
4th, clean：The powder that step 3 is obtained is placed in curved continuous box girder under 450~650W power, is returned with ether
Stream 15~30min of extraction, obtains dregs of a decoction I；Dregs of a decoction I extracts 15~30min with aether backflow；Obtain dregs of a decoction II；Dregs of a decoction II is used
Alcohol reflux extracts to obtain dregs of a decoction III；Dregs of a decoction III volatilizes solvent；
5th, extract：Obtained dregs of a decoction III is added to 10~15 times of water, is placed in ultrasonic refluxing extraction system, ultrasound is returned
Stream 1~2h of extraction, 0.5~1.5KW of ultrasonic power, 40~60 ° of Extracting temperature, obtains filtrate I；
6th, decolourize：0.1% activated carbon is added in filtrate I, decolourizes, obtains filtrate II；
7th, removing protein：Filtrate II is obtained into filtrate II I with Sevage method removing proteins；
8th, concentrate：Filtrate II I is put into microwave vacuum dryer, the span of Microwave Power Density and vacuum is
(2~8w/g) and (0.02~0.08Mpa), obtains lycium ruthenicum polysaccharide concentrated extracting solution；
9th, purify：By the D101 macroreticular resin wet method dress posts by pretreatment, above-mentioned crude extract is taken in 0.3~0.9ml/
Min loading different in flow rate, with the ethanol of 80% concentration with 0.4~1.5ml/min elution different in flow rate, eluent is closed
And concentrate, it is drying to obtain the higher polyoses extract of purity.
In the first step, if not cleaning carefully, silt dead leaf of doping etc. can influence extraction efficiency.
In second step, it is dried under low temperature, it is ensured that the preservation of polysaccharide active ingredient.
In 3rd step, superfine communication technique can make powder diameter small (typically below 15 μm) and be evenly distributed, ball
Degree and homogeneity are obviously improved, and bulk density and are significantly improved than surface, and plant cell wall breaking rate is high, in black fruit fructus lycii it is effective into
The dissolving and release for dividing (particularly refractory components) are accelerated, and the dissolution rate of active ingredient is accelerated.
In 5th step, the advantage of ultrasound and refluxing extraction is combined, extraction efficiency is higher.
In addition, for the consideration of control cost, low temperature drying, backflow, ultrasound in each step of the present invention, etc. be ripe
Technology, simple to operate, easy to get started.But compared with prior art, simplify extraction process and but obtained higher yield and product
The good product of matter.
Preferably, a kind of extracting method of lycium ruthenicum polysaccharide, it is characterised in that comprise the following steps：
1st, clear up：Black fruit fructus lycii 1kg is screened out into silt, cleaned with clear water；
2nd, dry：By the black fruit fructus lycii cleaned up at 20 DEG C low temperature drying 10h；
3rd, crush：The black fruit fructus lycii obtained to step 2 carries out ultramicro grinding, and it is 30 μm of powder to obtain particle size range；
4th, clean：The powder that step 3 is obtained is placed in curved continuous box girder under 450W power, is extracted with aether backflow
20min, obtain dregs of a decoction I；Dregs of a decoction I extracts 20min with aether backflow；Obtain dregs of a decoction II；Dregs of a decoction II is extracted with alcohol reflux
Dregs of a decoction III；Dregs of a decoction III volatilizes solvent；
5th, extract：Obtained dregs of a decoction III is added to 10 times of water, is placed in ultrasonic refluxing extraction system, ultrasound backflow carries
1h is taken, ultrasonic power 0.5KW, 40 ° of Extracting temperature, obtains filtrate I；
6th, decolourize：0.1% activated carbon is added in filtrate I, decolourizes, obtains filtrate II；
7th, removing protein：Filtrate II is obtained into filtrate II I with Sevage method removing proteins；
8th, concentrate：Filtrate II I is put into microwave vacuum dryer, Microwave Power Density 8w/g and 0.02Mpa, obtained black
Fruit LBP-X concentrated extracting solution；
9th, purify：By the D101 macroreticular resin wet method dress posts by pretreatment, above-mentioned crude extract is taken in 0.3ml/min stream
Fast loading, eluted with the ethanol of 80% concentration with 1.5ml/min flow velocity, eluent merged, concentrated, be drying to obtain purity compared with
High polyoses extract.
Preferably, a kind of extracting method of lycium ruthenicum polysaccharide, it is characterised in that comprise the following steps：
1st, clear up：Black fruit fructus lycii 1kg is screened out into silt, cleaned with clear water；
2nd, dry：By the black fruit fructus lycii cleaned up at 30 DEG C low temperature drying 10h；
3rd, crush：The black fruit fructus lycii obtained to step 2 carries out ultramicro grinding, and it is 35 μm of powder to obtain particle size range；
4th, clean：The powder that step 3 is obtained is placed in curved continuous box girder under 500W power, is extracted with aether backflow
25min, obtain dregs of a decoction I；Dregs of a decoction I extracts 25min with aether backflow；Obtain dregs of a decoction II；Dregs of a decoction II is extracted with alcohol reflux
Dregs of a decoction III；Dregs of a decoction III volatilizes solvent；
5th, extract：Obtained dregs of a decoction III is added to 9 times of water, is placed in ultrasonic refluxing extraction system, ultrasonic refluxing extraction
1.5h, ultrasonic power 1.5KW, 50 ° of Extracting temperature, obtain filtrate I；
6th, decolourize：0.1% activated carbon is added in filtrate I, decolourizes, obtains filtrate II；
7th, removing protein：Filtrate II is obtained into filtrate II I with Sevage method removing proteins；
8th, concentrate：Filtrate II I is put into microwave vacuum dryer, Microwave Power Density 5w/g and 0.05Mpa, obtained black
Fruit LBP-X concentrated extracting solution；
9th, purify：By the D101 macroreticular resin wet method dress posts by pretreatment, above-mentioned crude extract is taken in 0.5ml/min stream
Fast loading, eluted with the ethanol of 80% concentration with 1.0ml/min flow velocity, eluent merged, concentrated, be drying to obtain purity compared with
High polyoses extract.
Embodiment
The present invention is further detailed explanation by the following examples, but protection scope of the present invention is not by specific reality
Any restrictions of example are applied, but are defined in the claims.
Embodiment 1：The preparation of lycium ruthenicum polysaccharide
1st, clear up：Black fruit fructus lycii 1kg is screened out into silt, cleaned with clear water；
2nd, dry：By the black fruit fructus lycii cleaned up at 30 DEG C low temperature drying 10h；
3rd, crush：The black fruit fructus lycii obtained to step 2 carries out ultramicro grinding, and it is 35 μm of powder to obtain particle size range；
4th, clean：The powder that step 3 is obtained is placed in curved continuous box girder under 500W power, is extracted with aether backflow
25min, obtain dregs of a decoction I；Dregs of a decoction I extracts 25min with aether backflow；Obtain dregs of a decoction II；Dregs of a decoction II is extracted with alcohol reflux
Dregs of a decoction III；Dregs of a decoction III volatilizes solvent；
5th, extract：Obtained dregs of a decoction III is added to 9 times of water, is placed in ultrasonic refluxing extraction system, ultrasonic refluxing extraction
1.5h, ultrasonic power 1.5KW, 50 ° of Extracting temperature, obtain filtrate I；
6th, decolourize：0.1% activated carbon is added in filtrate I, decolourizes, obtains filtrate II；
7th, removing protein：Filtrate II is obtained into filtrate II I with Sevage method removing proteins；
8th, concentrate：Filtrate II I is put into microwave vacuum dryer, Microwave Power Density 5w/g and 0.05Mpa, obtained black
Fruit LBP-X concentrated extracting solution.
9th, purify：By the macroreticular resin wet method dress post by pretreatment, above-mentioned crude extract is taken on 0.5ml/min flow velocity
Sample, eluted with the ethanol of 80% concentration with 1.0ml/min flow velocity, eluent is merged, concentrates, it is higher to be drying to obtain purity
Polyoses extract.
The recovery rate for the lycium ruthenicum polysaccharide that embodiment 1 is extracted is far above the 1.37% of prior art for 3.89%, because
This extraction of present invention for lycium ruthenicum polysaccharide extract has preferable beneficial effect.
Embodiment 2：The preparation of lycium ruthenicum polysaccharide
1st, clear up：Black fruit fructus lycii 1kg is screened out into silt, cleaned with clear water；
2nd, dry：By the black fruit fructus lycii cleaned up at 20 DEG C low temperature drying 10h；
3rd, crush：The black fruit fructus lycii obtained to step 2 carries out ultramicro grinding, and it is 30 μm of powder to obtain particle size range；
4th, clean：The powder that step 3 is obtained is placed in curved continuous box girder under 450W power, is extracted with aether backflow
20min, obtain dregs of a decoction I；Dregs of a decoction I extracts 20min with aether backflow；Obtain dregs of a decoction II；Dregs of a decoction II is extracted with alcohol reflux
Dregs of a decoction III；Dregs of a decoction III volatilizes solvent.
5th, extract：Obtained dregs of a decoction III is added to 10 times of water, is placed in ultrasonic refluxing extraction system, ultrasound backflow carries
1h is taken, ultrasonic power 0.5KW, 40 ° of Extracting temperature, obtains filtrate I.
6th, decolourize：0.1% activated carbon is added in filtrate I, decolourizes, obtains filtrate II.
7th, removing protein：Filtrate II is obtained into filtrate II I with Sevage method removing proteins.
8th, concentrate：Filtrate II I is put into microwave vacuum dryer, Microwave Power Density 8w/g and 0.02Mpa, obtained black
Fruit LBP-X concentrated extracting solution.
9th, purify：By the macroreticular resin wet method dress post by pretreatment, above-mentioned crude extract is taken on 0.8ml/min flow velocity
Sample, eluted with the ethanol of 80% concentration with 0.8ml/min flow velocity, eluent is merged, concentrates, it is higher to be drying to obtain purity
Polyoses extract.
The recovery rate for the lycium ruthenicum polysaccharide that embodiment 2 is extracted is far above the 1.37% of prior art for 3.78%, because
This extraction of present invention for lycium ruthenicum polysaccharide extract has preferable beneficial effect.
Embodiment 3
Phenol-sulfuric acid and colorimetric method determines the polyoses content of different macroreticular resin type purifying
The preparation of 1 phenol liquid
Phenol 200g is taken, adds 0.2g aluminium flakes and 0.1g sodium acid carbonates to distill, collects 182 DEG C of cuts.5g is weighed, adds distilled water
100mL dissolvings are settled to, mixes and is transferred in brown bottle, are put standby in refrigerator.
The preparation of 2 standard liquids
The pure glucose 100.00mg of 105 DEG C of analyses for being dried to constant weight accurately is weighed, is dissolved with distilled water and held in 100mL
Constant volume in measuring bottle, 1mg/mL standard liquid is obtained, pipettes 2mL, 4mL, 6mL, 8mL, 10mL respectively, in 100mL volumetric flasks
Constant volume obtains 20 μ g/mL, 40 μ g/mL, 60 μ g/mL, 80 μ g/mL, 100 μ g/mL standard sugar juice.
The drafting of 3 standard curves
Take 2mL standards sugar juice to add the phenol solution 1mL that mass fraction is 5% to shake up, be rapidly added the 5mL concentrated sulfuric acids,
Shake up, after room temperature places 5min, boiling water bath 15min, be rapidly cooled to room temperature, length scanning is carried out with ultraviolet-visual spectrometer,
It is maximum absorption band to measure at 485nm, and blank control replaces sugar juice with distilled water.Obtain calibration curve equation.
4 Different Extraction Method polysaccharide determinations (except resin types are prepared using the method for embodiment 1)
Experimental result is as shown in table 1, and the D101 macroreticular resins that the present invention uses can significantly improve lycium ruthenicum polysaccharide
Recovery rate.
The contrast for the total starches content that the Different Extraction Method of table 1 obtains
| Extracting method | H-30 | XDA-1 macroporous absorbent resins | The equal hole decolorizing resins of XDA-7 | D101 macroreticular resins |
| Absorbance | 0.254 | 0.242 | 0.343 | 0.461 |
| Account for raw medicinal herbs content (%) | 1.49 | 1.29 | 1.61 | 3.78 |
Polyoses content under phenol-sulfuric acid and colorimetric method measure Different Extraction Method
The preparation of 1 phenol liquid
Phenol 200g is taken, adds 0.2g aluminium flakes and 0.1g sodium acid carbonates to distill, collects 182 DEG C of cuts.5g is weighed, adds distilled water
100mL dissolvings are settled to, mixes and is transferred in brown bottle, are put standby in refrigerator.
The preparation of 2 standard liquids
The pure glucose 100.00mg of 105 DEG C of analyses for being dried to constant weight accurately is weighed, is dissolved with distilled water and held in 100mL
Constant volume in measuring bottle, the standard liquid of 1mg mL~1 is obtained, pipettes 2mL, 4mL, 6mL, 8mL, 10mL respectively, in 100mL volumetric flasks
Middle constant volume obtains 20 μ g/mL, 40 μ g/mL, 60 μ g/mL, 80 μ g/mL, 100 μ g/mL standard sugar juice.
The drafting of 3 standard curves
Take 2mL standards sugar juice to add the phenol solution 1mL that mass fraction is 5% to shake up, be rapidly added the 5mL concentrated sulfuric acids,
Shake up, after room temperature places 5min, boiling water bath 15min, be rapidly cooled to room temperature, length scanning is carried out with ultraviolet-visual spectrometer,
It is maximum absorption band to measure at 485nm, and blank control replaces sugar juice with distilled water.Obtain calibration curve equation.
Polysaccharide determination (being prepared in addition to extracting method using the method for embodiment 1) under 4 different extractions
Experimental result：As shown in table 2, the extraction efficiency of lycium ruthenicum polysaccharide can be greatly improved using the present invention,
The content balance for the total starches that the Different Extraction Method of table 2 obtains
| Extracting method | Ultrasonic extraction | Refluxing extraction | Microwave Extraction | (present invention) |
| Absorbance | 0.249 | 0.243 | 0.350 | 0.459 |
| Account for raw medicinal herbs content (%) | 1.55 | 1.37 | 1.70 | 3.89 |
Embodiment 4
Research of the lycium ruthenicum polysaccharide to experimental mouse aging inoxidizability
The foundation and packet of 1 subacute aging animal model
Healthy kunming mice 96,25 ± 5g of body weight, female (♀) hero (♂) half and half, after feeding normal diet adaptation, claim on an empty stomach
Weight, eyeground vein blood sampling, survey in serum after MDA, be randomly divided into 6 groups, every group 16, body weight, serum MDA water between the groups of ♀ ♂ half and half
Flat no significant difference.Packet and administrations are as follows：
1 group：Normal group (Normal group)：Physiological saline+distilled water
2 groups：Aging model group (Control group)：D (+) galactolipin 200mg/kg+ distilled water
3 groups：Positive controls (positive group)：D (+) galactolipin 200mg/kg+ vitamin e1s 00mg/kg
4 groups：Polysaccharide experimental group (preparation of the method for embodiment 1) (Test group)：The black fruit Chinese hollys of D (+) galactolipin 200mg/kg+
Qi polyoses extract 400mg/kg
Above-mentioned lycium ruthenicum polysaccharide extract, obtained using the method for embodiment 1 is prepared.
2 medications
Lycium ruthenicum polysaccharide is dissolved in distilled water and controlled respectively at the daily morning to mouse stomach, gavage amount in 4ml/d
It is interior, continuous 8 weeks.It is injected intraperitoneally D (+) galactolipin 200mg/kg simultaneously, vitamin E is dissolved in salad oil that to be made into respective concentration same
Time gives gavage 8 weeks.
MDA, SOD, GSH-PX measure in 3 blood plasma
Mouse fasting be can't help into water 12 hours after eight weeks, 1.5~2.0ml of blood is taken from eyeground vein clump, prepare blood plasma, carried out
The measure of indices.When taking blood, with anticoagulant heparin, 4 DEG C, 3000r/min is centrifuged 15 minutes, is taken supernatant to be put into refrigerator and is treated
Survey.MDA, SOD, GSH-PX content use enzymatic assays by kit explanation in blood plasma.
The preparation of 4 liver homogenates and wherein MDA, SOD, GSH-PX measure
Mouse takes blood collare spondyloschisis to put to death from eyeground vein clump after being administered 8 weeks, wins mouse liver 0.5g, is put into grinding
In device, 4.5ml physiological saline is added, grinding is made 10% liver homogenate, vibration, stood overnight, 6 DEG C, 3000r/min centrifugations 30
Minute, supernatant is taken, the content of MDA, SOD, GSH-PX in kit measurement liver are determined using MDA, SOD, GSH-PX.Egg
Quantitatively determined in vain by kit explanation with biuret method.
Experimental result
Influence of 1 lycium ruthenicum polysaccharide to subacute aging model mice serum and Liver MDA
Serum MDA determination data is shown：Model group serum lipid peroxide MDA contents have obvious increasing than Normal group
Height (P<0.001), positive controls and lycium ruthenicum polysaccharide group can substantially reduce serum MDA level, more equal with model group
There is significant (P<0.01)；Hepatic tissue MDA determination datas are shown：It is more right than normal after model group Liver MDA modeling
Have according to a group MDA and substantially increase (P<0.001), positive controls, LBP-X group was given can substantially to reduce hepatic tissue MDA water
It is flat, with the more equal significance (P of model group<0.01), (table 3).
The lycium ruthenicum polysaccharide of table 3 to subacute aging model mice serum and Liver MDA influence (nmol/ml,
X±s)
| N | Serum MDA | Hepatic tissue MDA | |
| Normal group | 15 | 3.85±0.33 | 2.28±0.25 |
| Aging model group | 14 | 6.79±0.88*** | 4.17±0.35*** |
| Positive controls | 13 | 4.63±0.61△△△ | 3.56±0.35△△△ |
| Polysaccharide control group | 16 | 4.80±0.49△△△ | 2.68±0.14△△△ |
Note：Compared with Normal group：* * are P<0.001, * * is P<0.01, * P<0.05；With hyperlipidemia model group ratio
Compared with：△ is P<0.05, △ △ is P<0.01；Compared with positive controls：## is P<0.01
Influence serum of the 2 lycium ruthenicum polysaccharide groups to subacute aging model mouse superoxide dismutase (SOD) content
Measure SOD data are shown：There is obvious reduction (P than Normal group after model group SOD in serum content modeling<0.001) it is, positive right
It is horizontal according to group and polysaccharide the group significantly raised serum activity of SOD of energy, more it is statistically significant (P with model group<
0.05)；Hepatic tissue SOD determination datas are shown：There is obvious drop than Normal group SOD after model group hepatic tissue SOD content modelings
Low (P<0.001), positive controls, more sugar amount groups significantly raised hepatic tissue SOD activity levels of energy, it is more equal with model group
Significance (P<0.01), (table 4)
Influence of the lycium ruthenicum polysaccharide of table 4 to subacute aging model mouse superoxide dismutase (SOD) content
(nmol/ml, X ± s)
| N | SOD in serum | Hepatic tissue SOD | |
| Normal group | 15 | 330.45±55.67 | 266.38±33.71 |
| Aging model group | 14 | 260.03±58.03*** | 131.59±16.57*** |
| Positive controls | 13 | 310.55±31.98△△ | 193.7±19.74*△△ |
| Polysaccharide control group | 16 | 320.02±32.86△△ | 251.11±26.70△△ |
Note：Compared with Normal group：* * are P<0.001, * * is P<0.01, △ P<0.05, △ △ is P<0.01；
Compared with positive controls：## is P<0.01
Influence of 3 lycium ruthenicum polysaccharides to subacute aging model mice serum and hepatic tissue GSH-PX contents
Serum GSH-PX determination datas are shown：There is obvious drop than Normal group after model group serum GSH-PX content modelings
Low (P<0.001), positive controls and the lycium ruthenicum polysaccharide group significantly raised serum GSH-PX of energy are horizontal, compared with model group
Equal significance (P<0.001),；Hepatic tissue GSH-PX determination datas are shown：After model group hepatic tissue GSH-PX content modelings
There is obvious reduction (P than Normal group GSH-PX<0.001), the positive controls amount group significantly raised hepatic tissue GSH- of energy
PX activity levels, with the more equal significance (P of model group<0.001), (table 5).
The lycium ruthenicum polysaccharide of table 5 is to subacute aging model mice serum and the influence (nmol/ of hepatic tissue GSH-PX contents
Ml, X ± s)
| N | Serum GSH-PX | Hepatic tissue GSH-PX | |
| Normal group | 15 | 680.1±113.08 | 342.24±45.6 |
| Aging model group | 14 | 510.54±36.30*** | 266.69±69.69*** |
| Positive controls | 13 | 645.05±47.52△△△ | 257.49±23.22*△△ |
| Polysaccharide control group | 16 | 697.58±63.95** | 263.09±18.52* |
Note：Compared with Normal group：* * are P<0.001, * * is P<0.01, * P<0.05；With hyperlipidemia model group ratio
Compared with：△ is P<0.05, △ △ is P<0.01；Compared with positive controls：## is P<0.01
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not limited to the substantial technological content model of the present invention
Enclose, substantial technological content of the invention is broadly to be defined in the right of application, any technology that other people complete
Entity or method, if with the right of application defined in it is identical, also or a kind of equivalent change, will
It is considered as being covered by among the right.
Claims (5)
1. a kind of preparation method of lycium ruthenicum polysaccharide extract, it is characterised in that comprise the following steps：
1) clear up：Black fruit fructus lycii is screened out into silt, cleaned with clear water；
2) dry：By the black fruit fructus lycii cleaned up at 5~30 DEG C 10~24h of low temperature drying；
3) crush：The black fruit fructus lycii obtained to step 2 carries out ultramicro grinding, and it is 1~75 μm of powder to obtain particle size range；
4) clean：The powder that step 3 is obtained is placed in curved continuous box girder under 450~650W power, is carried with aether backflow
15~30min is taken, obtains dregs of a decoction I；Dregs of a decoction I extracts 15~30min with aether backflow；Obtain dregs of a decoction II；By dregs of a decoction II ethanol
Refluxing extraction obtains dregs of a decoction III；Dregs of a decoction III volatilizes solvent；
5) extract：Obtained dregs of a decoction III is added to 10~15 times of water, is placed in ultrasonic refluxing extraction system, ultrasound backflow carries
1~2h is taken, 0.5~1.5kW of ultrasonic power, 40~60 DEG C of Extracting temperature, obtains filtrate I；
6) decolourize：0.1% activated carbon is added in filtrate I, decolourizes, obtains filtrate II；
7) removing protein：Filtrate II is obtained into filtrate II I with Sevage method removing proteins；
8) concentrate：Filtrate II I is put into microwave vacuum dryer, the span of Microwave Power Density and vacuum is respectively
2~8W/g and 0.02~0.08MPa, obtain lycium ruthenicum polysaccharide concentrated extracting solution；
9) purify：Will by the D101 macroreticular resin wet method dress posts of pretreatment, take concentrated extracting solution in step 8 in 0.3~
0.9mL/min flow velocity loading, eluted with the ethanol of 80% concentration with 0.4~1.5mL/min flow velocity, eluent merged,
Concentration, is drying to obtain.
2. the preparation method of lycium ruthenicum polysaccharide extract according to claim 1, it is characterised in that the black fruit Chinese holly
The preparation method of Qi polyoses extract is：
1) clear up：Black fruit fructus lycii 1kg is screened out into silt, cleaned with clear water；
2) dry：By the black fruit fructus lycii cleaned up at 20 DEG C low temperature drying 10h；
3) crush：Ultramicro grinding is carried out to black fruit fructus lycii prepared by step 2, it is 30 μm of powder to obtain particle size range；
4) clean：The powder that step 3 is obtained is placed in curved continuous box girder under 450W power, is extracted with aether backflow
20min, obtain dregs of a decoction I；Dregs of a decoction I extracts 20min with aether backflow；Obtain dregs of a decoction II；Dregs of a decoction II is extracted with alcohol reflux
Dregs of a decoction III；Dregs of a decoction III volatilizes solvent；
5) extract：Obtained dregs of a decoction III is added to 10 times of water, is placed in ultrasonic refluxing extraction system, ultrasonic refluxing extraction 1h,
Ultrasonic power 0.5kW, 40 DEG C of Extracting temperature, obtains filtrate I；
6) decolourize：0.1% activated carbon is added in filtrate I, decolourizes, obtains filtrate II；
7) removing protein：Filtrate II is obtained into filtrate II I with Sevage method removing proteins；
8) concentrate：Filtrate II I is put into microwave vacuum dryer, Microwave Power Density and vacuum value be respectively 8W/g and
0.02MPa, obtain lycium ruthenicum polysaccharide concentrated extracting solution；
9) purify：By the D101 macroreticular resin wet method dress posts by pretreatment, the concentrated extracting solution in step 8 is taken in 0.3mL/
Min flow velocity loading, eluted with the ethanol of 80% concentration with 1.5mL/min flow velocity, eluent is merged, concentrated, drying is
.
3. the preparation method of lycium ruthenicum polysaccharide extract according to claim 1, it is characterised in that the black fruit Chinese holly
The preparation method of Qi polyoses extract is：
1) clear up：Black fruit fructus lycii 1kg is screened out into silt, cleaned with clear water；
2) dry：By the black fruit fructus lycii cleaned up at 30 DEG C low temperature drying 10h；
3) crush：The black fruit fructus lycii obtained to step 2 carries out ultramicro grinding, and it is 35 μm of powder to obtain particle size range；
4) clean：The powder that step 3 is obtained is placed in curved continuous box girder under 500W power, is extracted with aether backflow
25min, obtain dregs of a decoction I；Dregs of a decoction I extracts 25min with aether backflow；Obtain dregs of a decoction II；Dregs of a decoction II is extracted with alcohol reflux
Dregs of a decoction III；Dregs of a decoction III volatilizes solvent；
5) extract：Obtained dregs of a decoction III is added to 9 times of water, is placed in ultrasonic refluxing extraction system, ultrasonic refluxing extraction
1.5h, ultrasonic power 1.5kW, 50 DEG C of Extracting temperature, obtain filtrate I；
6) decolourize：0.1% activated carbon is added in filtrate I, decolourizes, obtains filtrate II；
7) removing protein：Filtrate II is obtained into filtrate II I with Sevage method removing proteins；
8) concentrate：Filtrate II I is put into microwave vacuum dryer, Microwave Power Density and vacuum value be respectively 5W/g and
0.05MPa, obtain lycium ruthenicum polysaccharide concentrated extracting solution；
9) purify：By the D101 macroreticular resin wet method dress posts by pretreatment, the concentrated extracting solution in step 8 is taken in 0.5mL/
Min flow velocity loading, eluted with the ethanol of 80% concentration with 1.0mL/min flow velocity, eluent is merged, concentrated, drying is
.
4. the preparation method of lycium ruthenicum polysaccharide extract according to claim 1, it is characterised in that the black fruit Chinese holly
The preparation method of Qi polyoses extract is：
1) clear up：Black fruit fructus lycii is screened out into silt, cleaned with clear water；
2) dry：By the black fruit fructus lycii cleaned up at 20 DEG C low temperature drying 15h；
3) crush：The black fruit fructus lycii obtained to step 2 carries out ultramicro grinding, and it is 1~75 μm of powder to obtain particle size range；
4) clean：The powder that step 3 is obtained is placed in curved continuous box girder under 500W power, is extracted with aether backflow
20min, obtain dregs of a decoction I；Dregs of a decoction I extracts 25min with aether backflow；Obtain dregs of a decoction II；Dregs of a decoction II is extracted with alcohol reflux
Dregs of a decoction III；Dregs of a decoction III volatilizes solvent；
5) extract：Obtained dregs of a decoction III is added to 14 times of water, is placed in ultrasonic refluxing extraction system, ultrasonic refluxing extraction
1.5h, ultrasonic power 1.0kW, 50 DEG C of Extracting temperature, obtain filtrate I；
6) decolourize：0.1% activated carbon is added in filtrate I, decolourizes, obtains filtrate II；
7) removing protein：Filtrate II is obtained into filtrate II I with Sevage method removing proteins；
8) concentrate：Filtrate II I is put into microwave vacuum dryer, the value of Microwave Power Density and vacuum is respectively 5W/g
And 0.05MPa, obtain lycium ruthenicum polysaccharide concentrated extracting solution；
9) purify：By the D101 macroreticular resin wet method dress posts by pretreatment, the concentrated extracting solution in step 8 is taken in 0.6mL/
Min flow velocity loading, eluted with the ethanol of 80% concentration with 0.9mL/min flow velocity, eluent is merged, concentrated, drying is
.
5. a kind of have improving eyesight nourishing liver, lipid-loweringing, anti-cancer, the lycium ruthenicum polysaccharide extract of anti-aging effects, it is characterised in that
Described lycium ruthenicum polysaccharide method for preparing extractive comprises the following steps：
1) clear up：Black fruit fructus lycii is screened out into silt, cleaned with clear water；
2) dry：By the black fruit fructus lycii cleaned up at 20 DEG C low temperature drying 23h；
3) crush：The black fruit fructus lycii obtained to step 2 carries out ultramicro grinding, and it is 1~75 μm of powder to obtain particle size range；
4) clean：The powder that step 3 is obtained is placed in curved continuous box girder under 600W power, is extracted with aether backflow
28min, obtain dregs of a decoction I；Dregs of a decoction I extracts 28min with aether backflow；Obtain dregs of a decoction II；Dregs of a decoction II is extracted with alcohol reflux
Dregs of a decoction III；Dregs of a decoction III volatilizes solvent；
5) extract：Obtained dregs of a decoction III is added to 12 times of water, is placed in ultrasonic refluxing extraction system, ultrasonic refluxing extraction 1~
2h, ultrasonic power 1.2kW, 45 DEG C of Extracting temperature, obtain filtrate I；
6) decolourize：0.1% activated carbon is added in filtrate I, decolourizes, obtains filtrate II；
7) removing protein：Filtrate II is obtained into filtrate II I with Sevage method removing proteins；
8) concentrate：Filtrate II I is put into microwave vacuum dryer, the value of Microwave Power Density and vacuum is respectively 7W/g
And 0.07MPa, obtain lycium ruthenicum polysaccharide concentrated extracting solution；
9) purify：By the D101 macroreticular resin wet method dress posts by pretreatment, the concentrated extracting solution in step 8 is taken in 0.8mL/
Min flow velocity loading, eluted with the ethanol of 80% concentration with 1.4mL/min flow velocity, eluent is merged, concentrated, drying is
.
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| CN111777695B (en) * | 2019-04-04 | 2021-11-26 | 中国科学院上海药物研究所 | Lycium ruthenicum polysaccharide LRP3-S1, and preparation method and application thereof |
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| CN111925457A (en) * | 2020-07-24 | 2020-11-13 | 哈工大机器人南昌智能制造研究院 | Method for preparing high-purity lycium ruthenicum polysaccharide |
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| CN103333268A (en) * | 2013-07-23 | 2013-10-02 | 草本创想生物科技（成都）有限公司 | Method for preparing lycium ruthenicum polysaccharide |
| CN103848921A (en) * | 2012-12-05 | 2014-06-11 | 中国科学院大连化学物理研究所 | Lyceum ruthenicum alkali-extracted polysaccharide and preparation method and application thereof |
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| CN103848921A (en) * | 2012-12-05 | 2014-06-11 | 中国科学院大连化学物理研究所 | Lyceum ruthenicum alkali-extracted polysaccharide and preparation method and application thereof |
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