CN110066313A - A kind of preparation method of high-purity Lycium chinense glycopeptide - Google Patents
A kind of preparation method of high-purity Lycium chinense glycopeptide Download PDFInfo
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- CN110066313A CN110066313A CN201910366105.1A CN201910366105A CN110066313A CN 110066313 A CN110066313 A CN 110066313A CN 201910366105 A CN201910366105 A CN 201910366105A CN 110066313 A CN110066313 A CN 110066313A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/145—Extraction; Separation; Purification by extraction or solubilisation
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
- C07K1/22—Affinity chromatography or related techniques based upon selective absorption processes
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/30—Extraction; Separation; Purification by precipitation
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/36—Extraction; Separation; Purification by a combination of two or more processes of different types
Abstract
The invention discloses a kind of preparation method of high-purity Lycium chinense glycopeptide, be fructus lycii or the crushing of fructus lycii waste residue, sieving, subcritical degreasing, high speed shear low temperature broken wall water are mentioned, high speed centrifugation, micro-filtration UF membrane, micro-filtration membrane concentration, inorganic/organic aqueous two-phase extraction, macroporous absorbent resin removal of impurities, ethanol precipitation, freeze-drying obtain high-purity Lycium chinense glycopeptide.Effective component polysaccharides content >=80% through detecting, in the extract.The present invention utilizes the micro-filtration UF membrane Lycium chinense glycopeptide of 30kD, the Lycium chinense glycopeptide that molecular weight is lower than 30kD is only obtained, and the Lycium chinense glycopeptide by molecular weight greater than 30kD discards, because the latter is metabolized slowly in vivo, causing largely to accumulate can induce " excessive internal heat ", and the bioactivity of this part Lycium chinense glycopeptide is low；Using nanofiltration membrane concentration, aqueous two-phase extraction, four kinds of macroporous absorbent resin removal of impurities, ethanol precipitation purification techniques, the purity and quality of obtained Lycium chinense glycopeptide ensure that.
Description
Technical field
The present invention relates to a kind of preparation process of Lycium chinense glycopeptide more particularly to a kind of preparation works of high-purity Lycium chinense glycopeptide
Skill belongs to active ingredient of Chinese herbs extraction and separation technology field.
Background technique
Fructus lycii, be matrimony vine of solanaceae plant (Lycium barbartum L.) dry mature fruit, name come from " mind
Agriculture book on Chinese herbal medicine warp ", have the history in more than 3000 years so far, is the earliest dual-purpose of drug and food substance for obtaining Ministry of Public Health's approval.Chinese medicine ancient books and records
How on the books to the medicinal efficacy of fructus lycii, modern pharmacology and natural product chemistry are studies have shown that Lycium chinense glycopeptide (LBP) is Chinese holly
One of main active of matrimony vine, be acid heteroglycan, polypeptide or protein composition complicated glycopeptide structure, content be 3% ~
9%.Studies have shown that Lycium chinense glycopeptide is a kind of water-solubility protein polysaccharide, 95% or more ingredient is glucosides compound, mainly includes Portugal
The ingredients such as grape sugar, arabinose, mannose, xylose, rhamnose and fructose, there are also various polysaccharide compounds.Lycium chinense glycopeptide has
Following physiologic function: 1) immunoregulation effect: Lycium chinense glycopeptide can be improved the phagocytic function of phagocyte, and it is thin to improve T lymph
The competence for added value of born of the same parents, while promoting cellular immunity and humoral immunity and then playing immunoregulation effect；2) delaying senility function: Chinese holly
The activity of glutathione and Sudismase can be improved in Qi glycopeptide, removes excessive free radical, reduces malonaldehyde and lipofuscin
Content to inhibit Apoptosis, is finally reached anti-aging, it is raw to improve body by inhibiting the expression of marrow gene c-myc
Order the purpose of power.It can also be by reducing the toxicity of amyloid beta albumen (Ab-peptide), reducing the excitability of glutamic acid
Neurotrosis caused by neurotrosis, reduction homocysteine (homocysteine) and reduction homocysteine cause
Neuron Apoptosis number of mechanisms play delay senescence effect；3) hypoglycemic effect: Lycium chinense glycopeptide can enhance impaired pancreas islet
Intracellular Sudismase activity, improves the oxidation resistance of islet cells, mitigates damage of the peroxide to cell, reduces
Malonaldehyde production quantity, to play hypoglycemic effect；4) liver protection: Lycium chinense glycopeptide is clear by increasing reactive oxygen species
Except enzymatic activity, body anti-oxidation protection mechanism is improved, oxygen radical is removed in time, rather than accumulated in intracellular,
Cause cytolipin peroxidating, leads to cell damage.By adjusting antioxidant enzyme, inhibition hepatocellular apoptosis protects liver cell,
And then reduce the generation of hepatic injury.5) antifatigue effect: polysaccharides can dramatically increase muscle glycogen, liver glycogen reserves amount, improve
Movement front and back blood lactic acid dehydrogenase total activity, reduction are moved through rear urea nitrogen content incrementss, accelerate blood urea nitrogen after movement
Clearance rate, to relieve fatigue and accelerate eliminate fatigue have the function of it is fairly obvious.6) antitumor action: polysaccharides passes through
Improve macrophages phagocytic capacity, the formation of splenocyte antibody, lymphocyte transformation reaction, cytotoxic T lymphocyte killing function
Lipid peroxidation energy number of ways can and be reduced to realize to kinds cancers such as liver cancer, colon cancer, cervical carcinoma, gastric cancer, leukaemia
Inhibiting effect；7) eye protection acts on: Lycium chinense glycopeptide acts on photosensory cell and retinal ganglial cells, can mitigate view
Retinal pigment is denaturalized patient's vision degeneration, has good effect to retinal pigment degeneration, glaucoma, diabetic retinopathy
Fruit.
Currently, the report that Lycium chinense glycopeptide extracts preparation method is more, it is broadly divided into direct Hot water extraction and organic solvent
Hot water extraction after (ether, petroleum ether, ethyl alcohol etc.) degreasing.These conventional methods not only can make Lycium chinense glycopeptide by high temperature shadow
It rings, space structure variation causes active decline, can also expend a large amount of organic solvent, cause environmental pollution.Therefore, research and development green
Efficient Lycium chinense glycopeptide industrialization method has great importance.CN104311690B reports a kind of extraction side of polysaccharides
Method, specifically: fructus lycii is crushed, 40 ~ 80 meshes are crossed, obtains wolfberry fruit powder, the water of solid-liquid ratio 1:5 ~ 1:10 is added, it is small to impregnate 8 ~ 16
When, active carbon decoloring is added, cellulase degradation is added, alkali protease and ficin are added, in microwave condition
Lower enzymatic hydrolysis is concentrated and is decolourized after enzymatic hydrolysis, is centrifuged, and is taken supernatant that alcohol precipitation is concentrated, then be centrifuged, is dried to obtain polysaccharides.This method
Obtained polysaccharides purity is no doubt higher, but soak extraction 8 ~ 18 hours, increase production cost, microwave and high temperature enzyme deactivation
Process all may cause Lycium chinense glycopeptide space structure and variation, cause activity and quality decline, active carbon decoloring can adsorb greatly
The Lycium chinense glycopeptide active constituent of amount.
Summary of the invention
The object of the present invention is to provide a kind of preparation methods of high-purity Lycium chinense glycopeptide.
The preparation method of Lycium chinense glycopeptide of the present invention, comprises the following steps that:
(1) it pre-processes: fructus lycii raw material being placed in pulverizer and is crushed, cross 40 ~ 60 meshes, obtain fructus lycii powder.Wherein fructus lycii raw material
For gained dried wolfberry slag after fructus lycii and medlar fresh fruit preparation fructus lycii magma；
(2) subcritical abstraction degreasing: fructus lycii powder is added in sub-critical extraction device, using organic solvent as spe medium, nitrogen
Under atmosphere is enclosed, extraction degreasing is carried out at 0.1 ~ 1MPa of extracting pressure, 10 ~ 30MHz of supersonic frequency；Extraction times are 1 ~ 5 time；Extraction
Taking the time is 10 ~ 60min；Fructus lycii slag is obtained after extraction.Wherein, spe medium is propane, butane, dimethyl ether etc.；Fructus lycii
The solid-liquid ratio of powder and spe medium is 1 ~ 3kg/L；
(3) high speed shear low temperature broken wall water mentions: above-mentioned fructus lycii slag being placed in multi-function extractor, deionized water is added, is cutting
0.5 ~ 2 h is extracted at 6000 ~ 12000 revs/min of revolving speed, temperature are cut lower than 60 DEG C, obtains Lycium chinense glycopeptide extraction mixture；
(4) high speed centrifugation: above-mentioned fructus lycii extraction mixed liquor is pumped into tube centrifuge, at rpm revs/min of revolving speed 10000 ~ 16000
High speed centrifugation under clock obtains Lycium chinense glycopeptide extracting solution A；
(5) micro-filtration UF membrane: above-mentioned Lycium chinense glycopeptide extracting solution A is pumped into the microfiltration membranes that molecular weight is 30kD, in pressure 2 ~ 6
Bar collects permeate at 30 ~ 50 DEG C of temperature, obtains Lycium chinense glycopeptide extracting solution B；
(6) micro-filtration membrane concentration: being pumped into the microfiltration membranes that molecular weight is 5kD for above-mentioned Lycium chinense glycopeptide extracting solution B, is 5 ~ 10 in pressure
Bar, temperature are to collect trapped fluid at 25 ~ 40 DEG C, obtain Lycium chinense glycopeptide extracting solution C；
(7) inorganic/organic aqueous two-phase extraction: being added inorganic/organic double-aqueous phase system in Lycium chinense glycopeptide extracting solution C, sufficiently shakes
It is shaken to mixed conjunction, 8000 ~ 10000 revs/min of lower high speed centrifugations collect lower phase (inorganic phase), obtain Lycium chinense glycopeptide solution D.Wherein inorganic/
In organic aqueous two-phase, inorganic phase is potassium dihydrogen phosphate, dipotassium hydrogen phosphate, ammonium sulfate；The mass fraction of inorganic phase is 15 ~ 30%；Have
Machine is mutually polyethylene glycol or ethylene glycol, and the mass fraction of organic phase is 15% ~ 30%；
(8) macroporous absorbent resin cleans: Lycium chinense glycopeptide solution D is pumped into large pore resin absorption column, loading flow velocity is 1.0 ~
2.0BV/ h collects absorption raffinate, obtains Lycium chinense glycopeptide solution E.Wherein, large pore resin absorption column includes 2 ~ 4 macroporous absorptions
Resin column wherein at least has 2 large pore resin absorption column series connection；The diameter height of large pore resin absorption column compares for 1:8 ~ 1:12；Macropore
Adsorbing resin is at least one of D101, LX-11, LX-60, LSA-10, LX-28, LX-38, AB-8；
(9) Lycium chinense glycopeptide solution E: being concentrated into 1.5 ~ 2 times of volumes of fructus lycii slag quality by ethanol precipitation, and ethyl alcohol is added, makes most
Whole concentration of alcohol is 50% ~ 70%, stands 10 ~ 12h, and filtering obtains Lycium chinense glycopeptide precipitating；
(10) it is freeze-dried: above-mentioned Lycium chinense glycopeptide pellet frozen is dry, obtain light yellow cotton-shaped Lycium chinense glycopeptide.
Lycium chinense glycopeptide prepared by the present invention is in light yellow or white class fluffy solid, polyoses content >=80%, particular exam side
Method is executed according to " polysaccharides " content assaying method in fructus lycii standard GB/T/T 18672-2014.
The advantages of the invention patent, is as follows:
1, the present invention utilizes the micro-filtration UF membrane Lycium chinense glycopeptide of 30kD, only Lycium chinense glycopeptide of the acquisition molecular weight lower than 30kD, and incite somebody to action
Lycium chinense glycopeptide of the molecular weight greater than 30kD discards, because the latter is metabolized slowly in vivo, causing largely to accumulate can induce " excessive internal heat ", and
And the bioactivity of this part Lycium chinense glycopeptide is low；
2, the present invention uses nanofiltration membrane concentration, aqueous two-phase extraction, four kinds of macroporous absorbent resin removal of impurities, ethanol precipitation purification techniques,
It ensure that the purity and quality of obtained Lycium chinense glycopeptide.
Specific embodiment
The preparation method of Lycium chinense glycopeptide of the present invention and the purity of product are further elaborated below by specific embodiment.
Embodiment 1
(1) 500g fructus lycii is successively cleaned with tap water, pure water, is dry, pulverize, cross 40 meshes；
(2) fructus lycii to pulverize and sieve is packed into and extracts cloth bag and fastens mouth, be placed in the closed extractor of subcritical abstraction equipment
In, feeding-passage is closed, is evacuated to -0.075MPa, after 10 ~ 20min of inflated with nitrogen, is pumped into according to solid-liquid ratio 1:1.5kg/L
The butane of 0.75L is heated to 35 DEG C, and setting extracting pressure 0.5MPa, supersonic frequency are to be extracted under 15KHz, extraction time
15min is extracted 3 times；After extraction, extractant is drawn in knockout drum.Vacuum pump is opened, by the pressure drop of knockout drum
To -0.06MPa, temperature is maintained at 35 DEG C, evaporates the butane in material, and the butane of gasification is compressed into liquid through diaphragm type compressor
State is back in solvent tank, closes vacuum pump, and the pressure of knockout drum is made to reach 0MPa, discharge fructus lycii oil, by fructus lycii slag from mentioning
It takes in tank and takes out；
(3) above-mentioned fructus lycii slag is set in a round bottom flask, deionized water 5.0L is added, open high speed shear system, revolving speed is set
It is 8000 revs/min, the temperature to water-bath rises to 60 DEG C of beginning timing, and 0.5h, closing high-speed shearing and water-bath are extracted in shearing
Pot heater.
(4) it is cooled to room temperature to temperature, above-mentioned fructus lycii extraction mixed liquor is poured into beaker, is opened with high speed low temperature centrifugal machine
Begin to be centrifuged, centrifuge speed is 8000 revs/min, merges supernatant and obtains Lycium chinense glycopeptide extracting solution A；
(5) continue that deionized water is pumped into Sai Duolisi cross-flow ultrafiltration film packet (molecular weight 30kD) cleaning systems with peristaltic pump
Then above-mentioned Lycium chinense glycopeptide extracting solution A is pumped into above-mentioned cleaned ultrafiltration membrane packet by 30min, collect through the saturating of microfiltration membranes packet
Liquid is crossed, after the flow velocity of permeate obviously slows down, stops peristaltic pump, obtains Lycium chinense glycopeptide extracting solution B；It is cleaned again with deionized water
Cross-flow ultrafiltration film packet is then pumped into dehydrated alcohol, closes liquid in-out mouth, saves；
(6) continue that deionized water is pumped into Sai Duolisi cross-flow ultrafiltration film packet (molecular weight 5kD) cleaning systems with peristaltic pump
30min, is then pumped into above-mentioned cleaned ultrafiltration membrane packet for above-mentioned Lycium chinense glycopeptide extracting solution B, and the flow velocity to permeate obviously subtracts
After slow, stop peristaltic pump, collects the Lycium chinense glycopeptide extracting solution C of ultrafiltration membrane packet retention；It is super to clean slipstream again with deionized water
Filter membrane packet is then pumped into dehydrated alcohol, closes liquid in-out mouth, saves；
(7) aqueous two-phase is formed with the water of 20% ammonium sulfate and 20% ethylene glycol and 60%, above-mentioned resulting Lycium chinense glycopeptide is extracted
Liquid C and ethylene glycol-ammonium sulfate double-aqueous phase system are added in separatory funnel, sufficiently shake mixing 20 minutes, 9000 revs/min of high speeds
Centrifugation 10 minutes collects lower phase, obtains Lycium chinense glycopeptide solution D；
(8) 50gD101 resin and 75gAB-8 resin are weighed, with 95% ethyl alcohol soaked overnight, be packed into large pore resin absorption column (if any
Bubble generates, and must drive bubble out of), successively cleaned with 95% ethyl alcohol and deionized water.Pre- place is added in above-mentioned Lycium chinense glycopeptide solution D
In the large pore resin absorption column managed, coutroi velocity 1BV/HR collects the absorption by resin layer from resin column outlet outflow
Raffinate, Lycium chinense glycopeptide solution E.After resin column is once cleaned with 60% second alcohol and water, the preservation of 95% ethyl alcohol is added.
(9) Lycium chinense glycopeptide solution E is concentrated under reduced pressure into 1L, ethyl alcohol is added, makes its final concentration of alcohol 50%, stood
Overnight, filtering obtains Lycium chinense glycopeptide precipitating；
(10) above-mentioned Lycium chinense glycopeptide pellet frozen is dry, obtain the cotton-shaped Lycium chinense glycopeptide of off-white color.It is marked according to fructus lycii country
" polysaccharides " content assaying method detects in quasi- GB/T 18672-2014, and the content of polysaccharides is 82.3%.
Embodiment 2
(1) the fructus lycii waste residue that 30kg preparation fructus lycii magma obtains dry, pulverize, crosses 40 meshes；
(2) the fructus lycii waste residue to pulverize and sieve is dispensed and extracts cloth bag and fastens mouth, packed 5 kilograms of each cloth is respectively implanted 3 Asias
In the closed extractor of critical extraction equipment, feeding-passage is closed, is evacuated to -0.075MPa, after 10 ~ 20min of inflated with nitrogen, beaten
Butane holding vessel is opened, the butane of 39L is passed through according to solid-liquid ratio 1:1.3kg/L, is heated to 35 DEG C, extracting pressure 0.5MPa is set,
Extraction time 20min, supersonic frequency 15KHz are extracted 5 times, after extraction, extractant are drawn in knockout drum.It opens
Vacuum pump is opened, the pressure of knockout drum is down to -0.06MPa, temperature is maintained at 35 DEG C, evaporates the butane in material, and will gasification
Butane be compressed into liquid through diaphragm type compressor, be back in solvent tank, close vacuum pump, reach the pressure of knockout drum
0MPa, discharge fructus lycii oil, fructus lycii slag is taken out from extractor；
(3) above-mentioned fructus lycii slag is taken out from cloth bag, is placed in multi-function extractor, deionized water 360L is added, open high speed
Cutting system, setting revolving speed are 4000 revs/min, and the feed liquid temperature being steam heated in multi-function extractor is 55 DEG C or so
Start timing, control temperature not above 60 DEG C, 1.0h is extracted in shearing, and closing high-speed shearing is logical in multi-function extractor interlayer
Cross tap water cooling；
(4) it is cooled to room temperature to temperature, above-mentioned fructus lycii extraction mixed liquor is pumped into the tubular type that revolving speed is 16000 revs/min and is centrifuged
Machine collects 300L Lycium chinense glycopeptide extracting solution A；
(5) above-mentioned Lycium chinense glycopeptide extracting solution A is pumped into the microfiltration membrane system of 30kD, the permeate for penetrating microfiltration membranes is collected, to saturating
Cross liquid flow velocity obviously slow down after, stop filtering, obtain 240L Lycium chinense glycopeptide extracting solution B；
(6) above-mentioned Lycium chinense glycopeptide extracting solution B is pumped into the microfiltration membrane system of 5kD, after the flow velocity of permeate obviously slows down, is stopped
It is pumped into, collects the 60L Lycium chinense glycopeptide extracting solution C of retention；
(7) aqueous two-phase is formed with the water of 20% ammonium sulfate and 20% ethylene glycol and 60%, above-mentioned resulting Lycium chinense glycopeptide is extracted
Liquid C and ethylene glycol-ammonium sulfate double-aqueous phase system are added in multi-function extractor, open stirring, are sufficiently mixed 20 minutes, static mistake
Night collects lower phase, obtains 60L Lycium chinense glycopeptide solution D；
(8) water that 1/3 volume is injected into resin container, by 100 kilograms D101 and 100 kilogram of LX-60 resin from roof manhole
Place is packed into tank, is intake from tank bottom, coutroi velocity about 2-4BV/HR, for about 1hr after drain since top.Backwash terminates
Afterwards, inlet valve is closed, static 0.5 hour, falling to resin just to be washed, it intakes from tank deck, flow velocity about 4-6BV/HR, from
Tank bottom discharge, continues just washing an end in.With the solution of the 4%NaOH of 4 times of resin volumes, resin is passed through with the flow velocity of 2BV/h
Layer after being all passed through, liquid level is put to retaining solution 2 hours at 30cm on resin layer, is then with demineralized water to pH 8
Only.It with 4% HCl solution of about 4 times of resin volumes, is passed through by above by the method for NaOH, then uses demineralized water, directly
To water outlet pH7.It is rinsed until when efflux uses one times of water dilution not muddy in test tube with 95% or more ethyl alcohol.Most
2hr is eluted with the speed of 2BV/hr with water afterwards, washing is to ethanol content less than 1%.Pre- place is added in above-mentioned Lycium chinense glycopeptide solution D
In the large pore resin absorption column managed, coutroi velocity 1BV/HR collects the absorption by resin layer from resin column outlet outflow
Raffinate, Lycium chinense glycopeptide solution E.After resin column is once cleaned with 60% second alcohol and water, the preservation of 95% ethyl alcohol is added；
(9) ethyl alcohol is added in Lycium chinense glycopeptide solution E, makes its final concentration of alcohol 50%, stands overnight, filtered, obtain fructus lycii sugar
Peptide precipitating；
(10) above-mentioned Lycium chinense glycopeptide pellet frozen is dry, obtain the cotton-shaped Lycium chinense glycopeptide of off-white color.It is marked according to fructus lycii country
" polysaccharides " content assaying method detects in quasi- GB/T 18672-2014, and the content of polysaccharides is 86.9%.
Claims (6)
1. a kind of preparation method of high-purity Lycium chinense glycopeptide, comprises the following steps that:
(1) it pre-processes: fructus lycii raw material being placed in pulverizer and is crushed, cross 40 ~ 60 meshes, obtain fructus lycii powder；
(2) subcritical abstraction degreasing: fructus lycii powder is added in sub-critical extraction device, using organic solvent as spe medium, nitrogen
Under atmosphere is enclosed, extraction degreasing is carried out at 0.1 ~ 1MPa of extracting pressure, 10 ~ 30MHz of supersonic frequency；Extraction times are 1 ~ 5 time；Extraction
Taking the time is 10 ~ 60min；Fructus lycii slag is obtained after extraction；
(3) high speed shear low temperature broken wall water mentions: above-mentioned fructus lycii slag being placed in multi-function extractor, deionized water is added, is cutting
0.5 ~ 2 h is extracted at 6000 ~ 12000 revs/min of revolving speed, temperature are cut lower than 60 DEG C, obtains Lycium chinense glycopeptide extraction mixture；
(4) high speed centrifugation: above-mentioned fructus lycii extraction mixed liquor is pumped into tube centrifuge, at rpm revs/min of revolving speed 10000 ~ 16000
High speed centrifugation under clock obtains Lycium chinense glycopeptide extracting solution A；
(5) micro-filtration UF membrane: above-mentioned Lycium chinense glycopeptide extracting solution A is pumped into the microfiltration membranes that molecular weight is 30kD, in pressure 2 ~ 6
Bar collects permeate at 30 ~ 50 DEG C of temperature, obtains Lycium chinense glycopeptide extracting solution B；
(6) micro-filtration membrane concentration: being pumped into the microfiltration membranes that molecular weight is 5kD for above-mentioned Lycium chinense glycopeptide extracting solution B, is 5 ~ 10 in pressure
Bar, temperature are to collect trapped fluid at 25 ~ 40 DEG C, obtain Lycium chinense glycopeptide extracting solution C；
(7) inorganic/organic aqueous two-phase extraction: being added inorganic/organic double-aqueous phase system in Lycium chinense glycopeptide extracting solution C, sufficiently shakes
It is shaken to mixed conjunction, 8000 ~ 10000 revs/min of lower high speed centrifugations collect lower phase, obtain Lycium chinense glycopeptide solution D；
(8) macroporous absorbent resin cleans: Lycium chinense glycopeptide solution D is pumped into large pore resin absorption column, loading flow velocity is 1.0 ~
2.0BV/ h collects absorption raffinate, obtains Lycium chinense glycopeptide solution E；
(9) Lycium chinense glycopeptide solution E: being concentrated into 1.5 ~ 2 times of volumes of fructus lycii slag quality by ethanol precipitation, and ethyl alcohol is added, makes most
Whole concentration of alcohol is 50% ~ 70%, stands 10 ~ 12h, and filtering obtains Lycium chinense glycopeptide precipitating；
(10) it is freeze-dried: above-mentioned Lycium chinense glycopeptide pellet frozen is dry, obtain light yellow cotton-shaped Lycium chinense glycopeptide.
2. a kind of preparation method of high-purity Lycium chinense glycopeptide as described in claim 1, it is characterised in that: pretreatment of raw material process
In, fructus lycii raw material is that fructus lycii and medlar fresh fruit prepare gained dried wolfberry slag after fructus lycii magma.
3. a kind of preparation method of high-purity Lycium chinense glycopeptide as described in claim 1, it is characterised in that: subcritical abstraction degreasers
In skill, spe medium is propane, butane, dimethyl ether.
4. a kind of preparation method of high-purity Lycium chinense glycopeptide as described in claim 1, it is characterised in that: subcritical abstraction degreasers
In skill, the solid-liquid ratio of fructus lycii powder and spe medium is 1 ~ 3kg/L.
5. a kind of preparation method of high-purity Lycium chinense glycopeptide as described in claim 1, it is characterised in that: inorganic/organic aqueous two-phase
In phase extraction process, inorganic phase is potassium dihydrogen phosphate, dipotassium hydrogen phosphate, ammonium sulfate；The mass fraction of inorganic phase is 15 ~ 30%；Have
Machine is mutually polyethylene glycol or ethylene glycol, and the mass fraction of organic phase is 15% ~ 30%.
6. a kind of preparation method of high-purity Lycium chinense glycopeptide as described in claim 1, it is characterised in that: macroporous absorbent resin removal of impurities
In technique, large pore resin absorption column includes 2 ~ 4 large pore resin absorption columns, wherein at least there is 2 large pore resin absorption column strings
Connection；The diameter height of large pore resin absorption column compares for 1:8 ~ 1:12；Macroporous absorbent resin is D101, LX-11, LX-60, LSA-10, LX-
28, at least one of LX-38, AB-8.
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| CN112451655A (en) * | 2020-12-10 | 2021-03-09 | 中国科学院兰州化学物理研究所 | Anti-fatigue composition and application thereof |
| CN113599495A (en) * | 2021-06-04 | 2021-11-05 | 爱尔眼科医院集团股份有限公司 | Composition and application thereof in preparation of medicine for treating retinitis pigmentosa |
| CN113599495B (en) * | 2021-06-04 | 2024-04-05 | 爱尔眼科医院集团股份有限公司 | Composition and application thereof in preparation of medicines for treating retinitis pigmentosa |
| CN113501859A (en) * | 2021-07-01 | 2021-10-15 | 宁夏杞奕农业发展有限公司 | Lycium barbarum glycopeptide purification and standing equipment and use method thereof |
| CN113616770A (en) * | 2021-08-10 | 2021-11-09 | 宁夏杞肽科技有限公司 | Use of a pharmaceutical mixture for the treatment or prevention of a disease caused by a coronavirus |
| CN113750217A (en) * | 2021-10-26 | 2021-12-07 | 宁夏杞肽科技有限公司 | Application of lycium barbarum glycopeptide |
| CN114052253A (en) * | 2021-11-19 | 2022-02-18 | 宁夏农林科学院枸杞科学研究所 | Method for preparing lycium barbarum zeaxanthin microcapsules by lycium barbarum polysaccharide composite wall material |
| CN114588250A (en) * | 2022-03-18 | 2022-06-07 | 宁夏杞肽科技有限公司 | Application of lycium barbarum glycopeptide in preparation of medicine for preventing or treating xerophthalmia |
| CN114588250B (en) * | 2022-03-18 | 2024-01-09 | 宁夏杞肽科技有限公司 | Application of lycium barbarum glycopeptide in preparing medicine for preventing or treating xerophthalmia |
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