CN107281212B - A kind of jujube complex polysaccharide composition and its preparation method and application - Google Patents
A kind of jujube complex polysaccharide composition and its preparation method and application Download PDFInfo
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- CN107281212B CN107281212B CN201710465590.9A CN201710465590A CN107281212B CN 107281212 B CN107281212 B CN 107281212B CN 201710465590 A CN201710465590 A CN 201710465590A CN 107281212 B CN107281212 B CN 107281212B
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- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
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Abstract
A kind of jujube complex polysaccharide composition of the present invention and its preparation method and application, the present invention uses jujube, Tremella fructification, the fruit of Chinese wolfberry, Radix Notoginseng respectively, Radix Notoginseng is the waste residue for extracting arasaponin, date polysaccharide, tremella polysaccharides, polysaccharides, neutral notoginseng polysaccharide are extracted, yield is respectively 6.5-6.9%, 12.0-21.0%, 2.8-4.4%, 0.32-0.42%.It is respectively 48-58%, 37-41%, 28-34%, 80-94% that H2SO4-anthrone method, which measures polyoses content,.This formula contributes to immunocompromised person and tumor patient strengthen immunity, improves the symptoms such as apathetic, physically weak weak, appetite reduction, nausea and vomiting, loose stool diarrhea, irritated insomnia, the harm for effectively mitigating chemicotherapy and performing the operation to tumour patient.
Description
Technical field
The invention belongs to field of medicaments, it is related to a kind of jujube complex polysaccharide composition and its preparation method and application.
Background technology
According to the website (http of the World Health Organization://www.who.int/mediacentre/factsheets/fs297/
Zh/) No. 297 display of cancer commentary in 2 months in 2015：The main reason for cancer is whole world morbidity and is dead.In 184 states
In family and area, Chinese Cancer Mortality occupies medium level on the upper side in general, and number of the infected accounts for about the whole world
21.8%.With aging of population aggravation and socio-economic development, industrialization, urbanization process are accelerated, environmental factor, life side
Formula constantly changes, and the situation is tense for China's tumour prevention and control.
In recent years, with the development of biotechnology, tumour immunotherapy becomes after tumor operation, radiotherapy, chemotherapy, and targeting is controlled
The means for being most hopeful successfully to cure cancer after treatment, this several years are even more to achieve major progress, currently, to target PD-1/
The Antybody therapy of PD-L1, it is a variety of pernicious in melanoma, non-small cell lung cancer, gastric cancer, liver cancer, carcinoma of urinary bladder and lymthoma etc.
Tumour all shows fine curative effect.Malignant tumour immunization therapy is given as breakthrough and is reported by Science, Nature
Road.
Neuroleptic Leukocytopenia, clinical rush leucocyte drug common at present, such as leucogen, shark are easily caused during chemotherapy of tumors
Liver alcohol etc., effect is slow, and effect is not notable.And though it is notable to inject the leukogenic effects such as class drug G-CSF, there are rebound phenomenon,
It will appear the side reactions such as ostalgia, fever when large dosage application, and price is relatively expensive.Therefore eager demand is curative for effect, valence of fine quality
Honest and clean, Small side effects drugs.
Chinese medicine removes the traditional idea for the treatment of disease (including tumour) from the angle for restoring body equilibrium between yin and yang, with modern medicine
Tumour immunotherapy realizes that the thinking of homeostasis plays the same tune on different musical instruments wonderful from immune function of human body is adjusted.Chinese medicine thinks
Why " heresy is gathered, and gas must be empty ", disease and evil can invade the imbalance of yin and yang that human body is mostly due to people itself, under immunity
Drop.By drug come correct negative and positive, strengthen the body resistance to consolidate the constitution be only cure the disease it is basic.Ginseng, Radix Astragali, ganoderma lucidum etc. are much rich in the benefit of polysaccharide
Beneficial class Chinese medicine is widely used in the various diseases including tumour.
Polysaccharide is widely present in animal, plant and microorganism.One of constituent as living matter, it joins extensively
With the various biological phenomenas of cell and the adjusting of physiology course, such as the transmission and impression of information between immunocyte, cell turns
The activities such as change, division and regeneration.Polysaccharide has antiviral, anti-infective, antitumor, anti-oxidant, radioresistance, hypoglycemic, liver protection, tune
Section is immune to wait physiological activity.A large amount of pharmacology and clinical research show that polysaccharide is a kind of immunomodulator, it can activate it is immune by
Body improves the immune function of body, realizes antitumor function.
The patent of invention that number of patent application is 201210532229.0, which discloses, provides a kind of strengthen immunity of contributing to
Health preserving health-care food formula and preparation method thereof.It includes the raw material of following weight part ratio：1000 parts of the fruit of Chinese wolfberry, sealwort 1000
Part, 1000 parts of Poria cocos, 1500 parts of Chinese yam, 1000 parts of mulberries, 1000 parts of jujube, 600 parts of donkey-hide gelatin, 600 parts of white fungus.It is prepared by the invention
Technique uses traditional water decoction extraction method, has a single function, the effect unobvious in terms of improving immunity.
Invention content
For overcome the deficiencies in the prior art, the present invention provides a kind of jujube complex polysaccharide composition of the present invention and its preparation
Methods and applications, it is to extract giving up for arasaponin that the present invention uses jujube, Tremella fructification, the fruit of Chinese wolfberry, Radix Notoginseng, Radix Notoginseng respectively
Slag, extraction date polysaccharide, tremella polysaccharides, polysaccharides, neutral notoginseng polysaccharide, yield is respectively 6.5-6.9%, 12.0-
21.0%, 2.8-4.4%, 0.32-0.42%, it is respectively 48-58%, 37-41%, 28- that H2SO4-anthrone method, which measures polyoses content,
34%, 80-94%.This formula contributes to immunocompromised person and tumor patient strengthen immunity, improves apathetic, physically weak weary
The symptoms such as power, appetite reduction, nausea and vomiting, loose stool diarrhea, irritated insomnia effectively mitigate chemicotherapy and operation to tumour patient
Harm.
The present invention provides the following technical solutions：
A kind of jujube complex polysaccharide composition, is made by mixing using plurality of raw materials extract, including date polysaccharide carries
Take object, Lycium barbarum polysaccharide extract and tremella polysaccharides extract, neutral notoginseng polysaccharide extract.
Preferably, include the extract of following parts by weight:0.5-5 parts of date polysaccharide extract, Lycium barbarum polysaccharide extract
0.5-5 parts, 0.5-5 parts of tremella polysaccharides extract, neutral 0-5 parts of notoginseng polysaccharide extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:4.5 parts of date polysaccharide extract, matrimony vine
1 part of polyoses extract, 4.5 parts of tremella polysaccharides extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:4.5 parts of date polysaccharide extract, matrimony vine
1 part of polyoses extract, 4.5 parts of tremella polysaccharides extract, neutral 1 part of notoginseng polysaccharide extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:2 parts of date polysaccharide extract, matrimony vine are more
1 part of sugar extract, 2 parts of tremella polysaccharides extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:2 parts of date polysaccharide extract, matrimony vine are more
1 part of sugar extract, 2 parts of tremella polysaccharides extract, neutral 2 parts of notoginseng polysaccharide extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:1 part of date polysaccharide extract, matrimony vine are more
1 part of sugar extract, 1 part of tremella polysaccharides extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:1 part of date polysaccharide extract, matrimony vine are more
1 part of sugar extract, 1 part of tremella polysaccharides extract, neutral 1 part of notoginseng polysaccharide extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:2 parts of date polysaccharide extract, matrimony vine are more
1 part of sugar extract, 1 part of tremella polysaccharides extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:2 parts of date polysaccharide extract, matrimony vine are more
1 part of sugar extract, 1 part of tremella polysaccharides extract, neutral 1 part of notoginseng polysaccharide extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:3 parts of date polysaccharide extract, matrimony vine are more
1 part of sugar extract, 1 part of tremella polysaccharides extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:3 parts of date polysaccharide extract, matrimony vine are more
1 part of sugar extract, 1 part of tremella polysaccharides extract, neutral 1 part of notoginseng polysaccharide extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:4 parts of date polysaccharide extract, matrimony vine are more
1 part of sugar extract, 1 part of tremella polysaccharides extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:4 parts of date polysaccharide extract, matrimony vine are more
1 part of sugar extract, 1 part of tremella polysaccharides extract, neutral 1 part of notoginseng polysaccharide extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:1 part of date polysaccharide extract, matrimony vine are more
2 parts of sugar extract, 1 part of tremella polysaccharides extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:1 part of date polysaccharide extract, matrimony vine are more
2 parts of sugar extract, 1 part of tremella polysaccharides extract, neutral 1 part of notoginseng polysaccharide extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:1 part of date polysaccharide extract, matrimony vine are more
3 parts of sugar extract, 1 part of tremella polysaccharides extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:1 part of date polysaccharide extract, matrimony vine are more
3 parts of sugar extract, 1 part of tremella polysaccharides extract, neutral 1 part of notoginseng polysaccharide extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:1 part of date polysaccharide extract, matrimony vine are more
4 parts of sugar extract, 1 part of tremella polysaccharides extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:1 part of date polysaccharide extract, matrimony vine are more
4 parts of sugar extract, 1 part of tremella polysaccharides extract, neutral 1 part of notoginseng polysaccharide extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:1 part of date polysaccharide extract, matrimony vine are more
1 part of sugar extract, 2 parts of tremella polysaccharides extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:1 part of date polysaccharide extract, matrimony vine are more
1 part of sugar extract, 2 parts of tremella polysaccharides extract, neutral 1 part of notoginseng polysaccharide extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:1 part of date polysaccharide extract, matrimony vine are more
1 part of sugar extract, 3 parts of tremella polysaccharides extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:1 part of date polysaccharide extract, matrimony vine are more
1 part of sugar extract, 3 parts of tremella polysaccharides extract, neutral 1 part of notoginseng polysaccharide extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:1 part of date polysaccharide extract, matrimony vine are more
1 part of sugar extract, 4 parts of tremella polysaccharides extract.
Any of the above-described scheme is preferably, and includes the extract of following parts by weight:1 part of date polysaccharide extract, matrimony vine are more
1 part of sugar extract, 4 parts of tremella polysaccharides extract, neutral 1 part of notoginseng polysaccharide extract.
Any of the above-described scheme is preferably, and further includes pharmaceutic adjuvant.Pharmaceutic adjuvant includes sucrose, starch, maltodextrin, bee
Honey, benzic acid sodium, ethyl hydroxy benzoate, magnesium stearate, protein sugar etc..
Any of the above-described scheme is preferably, and dosage form is any one in granule, tablet, capsule or oral solution.
The present invention also provides the preparation methods of above-mentioned jujube complex polysaccharide composition, include the following steps：
(1) extraction of date polysaccharide；
(2) extraction of polysaccharides；
(3) extraction of tremella polysaccharides；
(4) extraction of neutral notoginseng polysaccharide；
(5) said extracted object, packing are mixed according to weight proportion.
Any of the above-described scheme is preferably, and the extraction specific method of date polysaccharide is in the step (1)：Jujube crushes,
More times of amount water are added to impregnate, multi-reflow extraction discards filter residue merging filtrate, concentrates, ethyl alcohol alcohol precipitation is added, pumps supernatant and obtains
To precipitation and drying, dried powder is obtained.
Any of the above-described scheme is preferably, and the specific method of the alcohol precipitation is：The filtrate of separating and filtering, is concentrated into 1.06,
3 times of 95% ethyl alcohol is added to alcohol content 75%, the alcohol precipitation time is for 24 hours.
Any of the above-described scheme is preferably, and the specific method of the multi-reflow extraction is：The 8-10 times of water measured is added, just
Secondary refluxing extraction 5-8h, filters out solution, adds the 5-10 times of secondary water refluxing extraction 2-4h measured, filters out solution, add 4-6
The refluxing extraction 2-4h of water three times of amount again, discards filter residue merging filtrate.
Any of the above-described scheme is preferably, and the extraction specific method of polysaccharides is in the step (2)：Matrimony vine is added more
Amount water impregnates again, and multi-reflow extraction, merging filtrate, concentration, separating and filtering solution, addition ethyl alcohol alcohol precipitation, pumps after concentration
Clear liquid is precipitated and is dried, and dried powder is obtained.
Any of the above-described scheme is preferably, and the specific method of the multi-reflow extraction is：Water is measured in first 9-11 times of addition
It impregnates 12 hours, first refluxing extraction 5-8h filters out solution, adds the 4-10 times of water refluxing extraction 2-4h measured, secondary to filter out
Solution adds water that 3-5 times is measured refluxing extraction 2-4h three times, discards filter residue merging filtrate later.
Any of the above-described scheme is preferably, and the extraction specific method of tremella polysaccharides is in the step (3)：White fungus is added more
Amount water impregnates again, multi-reflow extraction, merging filtrate, concentration, and addition ethyl alcohol alcohol precipitation pumps supernatant and precipitated and dried,
Obtain dried powder.
Any of the above-described scheme is preferably, and the specific method of the multi-reflow extraction is：Water is measured in first 48-52 times of addition
It impregnates 12 hours, first refluxing extraction 5-8h filters out solution, adds the 30-40 times of water refluxing extraction 3-5h measured, secondary to filter out
Solution, merging filtrate.
Any of the above-described scheme is preferably, and neutral notoginseng polysaccharide is by extracting arasaponin in the step (4)
Waste residue, through water extract-alcohol precipitation, DEAE Sepharose Fast Flow anion-exchange chromatographies gained after purification.
Any of the above-described scheme is preferably, and packing further includes before that wheat is added into mixed extract in the step (5)
After mixing, 95% appropriate amount of ethanol is added in bud dextrin, crosses the granulation of 14 mesh screens, and drying is crossed 20 mesh screens and arranged, packing.Point
It can also can again be dispensed after tabletting with direct packaging when dress.
The present invention also provides above-mentioned jujube complex polysaccharide composition improve immunity, improve it is apathetic, physically weak it is weak,
Application in appetite reduction, nausea and vomiting, loose stool diarrhea, irritated insomnia.
The present invention also provides above-mentioned jujube complex polysaccharide compositions to be applied to health food.
The present invention also provides above-mentioned jujube complex polysaccharide compositions to be applied to cosmetics.
The present invention also provides above-mentioned jujube complex polysaccharide compositions to be applied to daily chemical product.Jujube in this formula is sandlwood
The ripening fruits of section's plant jujube.Chinese medicine theory thinks that jujube has the effects that tonify deficiency benefit health, nourishing blood and tranquilization, strengthening the spleen and stomach,
It is the good health nutrient of the patients such as weakness of the spleen and the stomach, insufficiency of vital energy and blood, lassitude, insomnia and dreamful sleep.Date polysaccharide has a variety of
Pharmacological activity, date polysaccharide are the main actives of jujube qi-supplementing, blood-engendering, and date polysaccharide is equal to the various active oxygens in human body
It is turned into antioxygen, repairs hepatic injury, antifatigue effect, immunocompetence, improves enteron aisle, inhibits tumour cell and promote the suction of calcium
It receives.
Matrimony vine is China's tradition rare traditional Chinese medicine, is known as " red treasured " laudatory title, rich in plant polyose, protein, vitamin etc.
Nutritional ingredient.Polysaccharides is a kind of water-soluble polysaccharide obtained by being extracted in matrimony vine.The clear polysaccharide system proteoglycan, by
6 kinds of arabinose, glucose, galactolipin, mannose, xylose, rhamnose monosaccharide components compositions.With promote T, B, CTL, NK and
The immune functions such as macrophage promote the cell factors such as IL-2, IL-3 and TNF β to generate.Enhance lotus knurl, chemotherapy and radiation injury
Immune function of mice and the effect for adjusting neuro-endocrine-immunoregulatory adjusting (NIM) network.
White fungus (Tremella fuciformisBerk), is commonly called as tremella, and it is more to obtain white fungus in fructification
(Tremellam) be a kind of acid heteroglycan, backbone structure be by the mannosan of α-(1 → 3) glucosides key connection, branch by
Glucuronic acid and xylose composition.For Basidiomycetes polysaccharide para-immunity reinforcing agent, having improves body's immunity and is promoted white thin
The effect of born of the same parents.It is remarkably improved the phagocytic function of experimental animal reticuloendothelial cell, has and promotes nospecific immunity effect, improve
The level of immunoglobulin, total complement of serum.It can prevent and treat and be pressed down by radiation and marrow caused by antitumor drug (such as CTX)
System, can also promote the synthesis of liver internal protein.
The present invention, using water extraction and alcohol precipitation method, obtains neutrality using the Radix Notoginseng waste residue for extracting notoginsenoside in producing as raw material
Radix Notoginseng roughly polysaccharide purifies Radix Notoginseng Thick many candies, in obtaining using DEAE Sepharose Fast Flow anion-exchange chromatographies
Property notoginseng polysaccharide PNPS I, study its influence to Periodontal ligament stem cell, it was demonstrated that neutral notoginseng polysaccharide PNPS I promote periodontal
The effect of film stem cell is more apparent, is protruded improving the immunity of the human body aspect effect.
A kind of jujube complex polysaccharide composition of present invention offer and its preparation method and application uses jujube, white fungus respectively
Entity, the fruit of Chinese wolfberry, Radix Notoginseng, Radix Notoginseng are the waste residue for extracting arasaponin, extraction date polysaccharide, tremella polysaccharides, polysaccharides,
Neutral notoginseng polysaccharide, yield are respectively 6.5-6.9%, 12.0-21.0%, 2.8-4.4%, 0.32-0.42%, sulfuric acid-anthrone
It is respectively 48-58%, 37-41%, 28-34%, 80-94% that method, which measures polyoses content,.This formula contributes to immunocompromised person
And tumor patient strengthen immunity, improve apathetic, physically weak weak, appetite reduction, nausea and vomiting, loose stool diarrhea, irritated mistake
The symptoms such as dormancy, the harm for effectively mitigating chemicotherapy and performing the operation to tumour patient.
Description of the drawings
Fig. 1 is the High Performance Gel Permeation chromatogram of neutral notoginseng polysaccharide PNPSI.
Specific implementation mode
In order to further appreciate that the technical characteristic of the present invention, the present invention is explained in detail with reference to specific embodiment
It states.Embodiment only has illustrative effect to the present invention, without the effect of any restrictions, those skilled in the art
The modification for any unsubstantiality made on the basis of the present invention should all belong to the scope of protection of the present invention.
Embodiment 1
The present invention is made of following raw materials, date polysaccharide extract, Lycium barbarum polysaccharide extract, tremella polysaccharides extract, in
Property notoginseng polysaccharide extract and pharmaceutically acceptable pharmaceutic adjuvant composition.Pharmaceutic adjuvant include sucrose, starch, maltodextrin,
Honey, benzic acid sodium, ethyl hydroxy benzoate, magnesium stearate, protein sugar etc..
The jujube complex polysaccharide composition of the present invention, including date polysaccharide extract 0.5-5kg, Lycium barbarum polysaccharide extract
0.5-5kg and tremella polysaccharides extract 0.5-5kg, neutral notoginseng polysaccharide extract 0-5kg, are added suitable for pharmaceutic adjuvant, mixing,
Add 95% appropriate amount of ethanol, cross the granulation of 14 mesh screens, drying is crossed 20 mesh screens and arranged, is distributed into 1000 bags.Packing is prepared into
The peroral dosage forms such as granula, tablet, capsule, oral solution.
In 1 bag of jujube complex polysaccharide composition of the invention, adult's recommendation dosage is：One day 2-3 times, one time 1-2 bags.
The preparation method of jujube complex polysaccharide composition of the present invention, including following step：
(1) extraction of date polysaccharide extract and assay
400 kilograms of jujube is weighed, is crushed, 8 times of amount water are added and impregnate 12 hours, refluxing extraction 8h, filter out solution, medicinal material is again
The water refluxing extraction 4h of 5 times of amounts is added, filters out solution, medicinal material adds the water refluxing extraction 4h of 4 times of amounts, discards filter residue and merges filter
Liquid is concentrated into 1.06,3 times of 95% ethyl alcohol is added to alcohol content 75%, the alcohol precipitation time is for 24 hours that pumping supernatant must precipitate and do
It is dry, obtain 27.60 kilograms of dried powder, yield 6.90%.It is appropriate that precision weighs powder, and H2SO4-anthrone method measures date polysaccharide
Content is 57.17%.
(2) extraction of Lycium barbarum polysaccharide extract and assay
400 kilograms of matrimony vine is weighed, 10 times of amount water are added and impregnate 12 hours, refluxing extraction 8h, filter out solution, medicinal material adds
The water refluxing extraction 4h of 5 times of amounts filters out solution, and medicinal material adds the water refluxing extraction 4h of 4 times of amounts, discards filter residue merging filtrate,
Density is concentrated into 1.06, it is 75% that 3 times of 95% ethyl alcohol to amount of alcohol, which is added, and the alcohol precipitation time is for 24 hours that pumping supernatant must precipitate
And it is dry, obtain 11.20 kilograms of dried powder, yield 2.80%.It is appropriate that precision weighs powder, and H2SO4-anthrone method measures matrimony vine
Polyoses content is 33.50%.
(3) extraction of tremella polysaccharides extract and assay
400 kilograms of white fungus is weighed, 50 times of amount water are added and impregnate 12 hours, refluxing extraction 8h, filter out solution, medicinal material adds
The water refluxing extraction 4h of 30 times of amounts, filters out solution, and merging filtrate is concentrated into density to 1.06,3 times of 95% ethyl alcohol is added to ethyl alcohol
Amount is 75%, and the alcohol precipitation time is for 24 hours that pumping supernatant must precipitate and dry, and obtain 47.84 kilograms of dried powder, and yield is
11.96%.It is appropriate that precision weighs powder, and it is 37.00% that H2SO4-anthrone method, which measures tremella polysaccharides content,.
(4) extraction of neutral notoginseng polysaccharide and assay
(4.1) extraction of Radix Notoginseng Thick many candies：The waste residue 1kg for extracting arasaponin is weighed, 10 times of amounts are added for the first time
Water, boiling water bath boil 8h, collect filtrate, second of addition, 5 times of amount water, boiling water bath 8h.Merge filtrate twice, 4500rpm centrifugations
5min takes supernatant.80 DEG C of spin concentrations are stood for 24 hours to the 1/8 of original volume, and centrifugation abandons precipitation, is slowly added to the anhydrous second of three times
Alcohol, stirring while adding, 4 DEG C of placement 12h filter to obtain precipitation.75% ethyl alcohol washing precipitation for several times, is used appropriate after ethyl alcohol volatilization is done
Supernatant is collected in water dissolution, centrifugation, and three times absolute ethyl alcohol is added, and 4 DEG C of placement 12h are filtered to obtain and be precipitated, 75% ethyl alcohol washing precipitation,
50 DEG C of baking oven drying are to get Radix Notoginseng Thick many candies.
(4.2) DEAE Sepharose Fast Flow anion-exchange chromatographies purify Thick many candies
(4.2.1) fills column：Wet method dress post, pillar fill in cotton, and distilled water is added in column and keeps a bit of liquid level, glass
Stick pressing cotton dispels bubble.It is poured slowly into column along column wall with glass bar guiding homogenate, bubble cannot be generated.Open pillar
Liquid outlet makes gel free settling in column, washes with water chromatographic column edge.There cannot be tomography when filling column, then use distilled water
Balance, column specification are 2.4 × 40cm.
(4.2.2) loading：Radix Notoginseng Thick many candies are dissolved in a small amount of distilled water, are configured to 30mg/mL, 4500rpm centrifugations
10min, 0.45 μm of filter filtering of supernatant, loading.When loading, chromatographic column liquid outlet is opened, it is to be distilled to move to column bed under water
When surface, liquid outlet is closed, notoginseng polysaccharide solution is added along column wall with dropper, opens liquid outlet, sample liquid is allowed all to flow into
A small amount of distilled water flushing column wall is used in combination in column interior, then row elution.
(4.2.3) is eluted：It is eluted with water, about 5 times of column volumes of elution, flow velocity 0.8mL/min, often pipe meets 2mL, and decompression is dense
It is reduced to 1/10 volume.
(4.2.4) dialyses：Bag filter is cut into every section of about 10cm, is put in distilled water and boils 30min, boiled again after washing three times
Boil 10min, distilled water wash clean.Bag filter can use three times, use 2%NaHCO for the second time3After being boiled with 1mmol/L EDTA
Cleaning.Respectively by bag filter of the eluent after concentration loaded on molecular weight 14000, about 1/2 volume is clamped with dialysis clamp and is dialysed
Bag both ends, are put in distilled water the 36h that dialyses, and a water is changed per 4h.
(4.2.5) is freeze-dried：Eluent is collected in 50mL centrifuge tubes, fills about 16mL, -40 DEG C of pre-freeze 12h are put in cold
Lyophilizer 36h is added about 1mL distillation water dissolutions, shakes up after taking-up.- 40 DEG C of pre-freeze 12h, are put in freeze drier for 24 hours.It takes
Sealing preserves in drier after going out.Anthrone-sulfuricacid method measures neutrality notoginseng polysaccharide PNPS I, PNPS I (neutral notoginseng polysaccharide)
Content 93.6%, yield 28.9%, number-average molecular weight 11170, weight average molecular weight 31590, polydispersity coefficient D (Mw/
Mn it is) 2.828, the High Performance Gel Permeation chromatogram of neutral notoginseng polysaccharide PNPS I is as shown in Figure 1.
(4.3) mtt assay measures influence of the neutral notoginseng polysaccharide to Periodontal ligament stem cell in-vitro multiplication
Using regular growth cultivation culture cell.The Periodontal ligament stem cell for taking 7 generation exponential phases is adjusted thin after counting
Born of the same parents' concentration, is inoculated in 96 orifice plates, 100/hole, per hole 90 μ l, CO2Incubator culture adds neutral notoginseng polysaccharide respectively after 12 hours
(PNPS I) and each 10 μ l of Radix Notoginseng Thick many candies solution, make to act on final concentration of 6.72mg/ml, 4.48mg/ml of cell,
2.24mg/ml, 1.12mg/ml, 560ug/ml, 280ug/ml, 140ug/ml, 70ug/ml, each concentration set 6 parallel holes, cloudy
Property control be isometric MEM-ALPHA culture solutions, add 10 μ l of physiological saline.After 48 hours, add 20 μ l of MTT solution (5mg/ml) every
Culture 4 hours is continued in hole, and 150 μ lDMSO are added per hole and fully dissolve, the A570 values in each hole are measured with microplate reader.It calculates different
The OD values of the A570 of concentration, experimental result are shown in Table 1：
Influence experimental result (n=6) of 1 notoginseng polysaccharide of table to Periodontal ligament stem cell in-vitro multiplication
Note：*P<0.05；**P<0.01
By data in table 1 it is found that neutrality notoginseng polysaccharide PNPS I is in 35~1120ug/ml of concentration, to Periodontal ligament stem cell
With apparent Effect of promoting growth；Relative to Radix Notoginseng Thick many candies, increasings of the neutral notoginseng polysaccharide PNPS I to Periodontal ligament stem cell
It is more notable to grow facilitation.
2 jujube complex polysaccharide granule of embodiment
Add maltodextrin appropriate, mixing adds 95% appropriate amount of ethanol, crosses the granulation of 14 mesh screens, and it is whole to cross 20 mesh screens for drying
Reason, is distributed into 1000 bags.
Date polysaccharide extract, Lycium barbarum polysaccharide extract, tremella polysaccharides extract, neutral notoginseng polysaccharide in the present embodiment
The preparation method of extract is and embodiment 1 is identical.
3 jujube complex polysaccharide tablet of embodiment
Add maltodextrin appropriate, mixing adds 95% appropriate amount of ethanol, crosses the granulation of 14 mesh screens, and it is whole to cross 20 mesh screens for drying
Reason, tabletting, film coating, packing.
Date polysaccharide extract, Lycium barbarum polysaccharide extract, tremella polysaccharides extract, neutral notoginseng polysaccharide in the present embodiment
The preparation method of extract is and embodiment 1 is identical.
4 jujube complex polysaccharide tablet of embodiment
Add maltodextrin appropriate, mixing adds 95% appropriate amount of ethanol, crosses the granulation of 14 mesh screens, and it is whole to cross 20 mesh screens for drying
Reason, tabletting, film coating, packing.
Date polysaccharide extract, Lycium barbarum polysaccharide extract, tremella polysaccharides extract, neutral notoginseng polysaccharide in the present embodiment
The preparation method of extract is and embodiment 1 is identical.
5 jujube complex polysaccharide granule of embodiment
Add maltodextrin appropriate, mixing adds 95% appropriate amount of ethanol, crosses the granulation of 14 mesh screens, and it is whole to cross 20 mesh screens for drying
Reason, is distributed into 1000 bags.
Date polysaccharide extract, Lycium barbarum polysaccharide extract, tremella polysaccharides extract, neutral notoginseng polysaccharide in the present embodiment
The preparation method of extract is and embodiment 1 is identical.
6 jujube complex polysaccharide granule of embodiment
Add maltodextrin appropriate, mixing adds 95% appropriate amount of ethanol, crosses the granulation of 14 mesh screens, and it is whole to cross 20 mesh screens for drying
Reason, is distributed into 1000 bags.
Date polysaccharide extract, Lycium barbarum polysaccharide extract, tremella polysaccharides extract, neutral notoginseng polysaccharide in the present embodiment
The preparation method of extract is and embodiment 1 is identical.
Embodiment 7
Jujube complex polysaccharide composition treats leukopenic observation of curative effect after malignant tumor chemotherapy
7.1 data and method
7.1.1 data
Caused Neuroleptic Leukocytopenia patient (is demonstrate,proved by pathology after choosing in June, 2015~2016 year malignant tumor chemotherapy in June
It is real), exclude to use other shengbai drugs person, KPS scorings >=60 points, total 50 are included in this research.It is randomly divided into observation
Each 10 with control group, in observation group man 4, female 6, average age (54.2 ± 7.3) year, breast cancer 4, cervical cancer 2 example,
Colorectal cancer 2, lung cancer 2.
In control group A man 5, female 5, average age be (53.9 ± 10.6) year, breast cancer 2, liver cancer 3, large intestine
Cancer 1, lung cancer 3, gastric cancer 1.In control group B man 6, female 4, average age be (54.1 ± 9.3) year, breast cancer 2,
Liver cancer 3, colorectal cancer 1, lung cancer 2, gastric cancer 2.Man 5 in control group C, female 5, average age are (53.6 ± 10.9)
Year, mammary cancer 1 example, liver cancer 3, colorectal cancer 2, lung cancer 4.In control group D man 6, female 4, average age be (53.7 ±
10.8) year, breast cancer 3, liver cancer 4, gastric cancer 3.
Four groups in gender composition, average age, KPS scorings, tumour classification there are no significant difference (P>0.05), having can
Compare property.
7.1.2 therapy
Observation group give prepared in embodiment 2 jujube complex polysaccharide composition (date polysaccharide 4.5g, polysaccharides 1.0g,
Tremella polysaccharides 0.5g, neutrality notoginseng polysaccharide 0.5g) each 1 time sooner or later, primary two bags, it even served 3 weeks.
Control group A gives batyl alcohol 100mg, and leucogen 20mg 3 times/d, even served 3 weeks.Close observation is various simultaneously during treatment
Send out the generation of disease.
Control group B takes product prepared by the patent of invention embodiment 1 that number of patent application is 201210532229.0, daily
Twice, three weeks be even served.
The jujube complex polysaccharide granule that control group C takes the preparation of embodiment 5 twice daily even served three weeks.
The jujube complex polysaccharide granule that control group D takes the preparation of embodiment 6 twice daily even served three weeks.
7.1.3 observation index and the standard of curative effect evaluation
Periphery blood routine is checked during treatment 2 times a week.According to the Ministry of Public Health《Clinical drug studies guideline》Middle formulation
Increasing leukocyte criterion of therapeutical effect, it is effective：WBC Zong Shuos >4.0 × 109/L can maintain curative effect to being discontinued 1 week, and clinical symptoms significantly subtract
Light or disappearance；Effectively：WBC Zong Shuos <4.0 × 109/L can maintain curative effect to stopping than rising (0.5-1.0) × 109/L before treatment
Medicine 1 week, clinical symptoms improve；In vain：WBC sum Zeng Gaos <0.5 × 109/L, clinical symptoms are without improvement.
7.2 treatment results
Data result is shown in Table 2 after treatment, and the treatment of observation group relative comparison group A, control group B, control group C, control group D are imitated
Fruit is more notable.Observation group has no adverse reaction.
Effect data after table 2 is treated
Clinical rush leucocyte drug common at present, such as leucogen, batyl alcohol, effect is slow, and effect is not notable.And it injects
Though the leukogenic effects such as class drug G-CSF are notable, there are rebound phenomenon, large dosage will appear ostalgia, fever etc. when applying secondary anti-
It answers, and price is relatively expensive.Therefore eager demand drug curative for effect, of high quality and at a reasonable price, Small side effects.
By the data in table 2 it is found that occurring the patient of leucocyte reduction after chemotherapy, jujube prepared by Application Example 2 is multiple
After closing polysaccharide composition, the deficiency of spleen and stomach symptoms such as soreness and weakness of waist and knees, dizzy, spiritlessness and weakness, deficiency of food are indigestion and loss of appetite have clear improvement,
KPS scorings before medication compared with being significantly improved.Effect unobvious after control group A, control group B and control group C, control group D treatments.
The above results show that jujube complex polysaccharide composition can treat leucocyte caused by chemotherapy of tumors to a certain extent
It reduces, to ensure being normally carried out for chemotherapy, extends patient survival.
Claims (17)
1. a kind of jujube complex polysaccharide composition, is made by mixing using plurality of raw materials extract, it is characterised in that：By following
The extract of parts by weight is made:0.5-5 parts of date polysaccharide extract, 0.5-5 parts of Lycium barbarum polysaccharide extract, tremella polysaccharides extract
0.5-5 parts, neutral 0-5 parts of notoginseng polysaccharide extract.
2. jujube complex polysaccharide composition according to claim 1, it is characterised in that：Include the extraction of following parts by weight
Object:4.5 parts of date polysaccharide extract, 1 part of Lycium barbarum polysaccharide extract, 4.5 parts of tremella polysaccharides extract, neutral notoginseng polysaccharide extraction
1 part of object.
3. jujube complex polysaccharide composition according to claim 1, it is characterised in that：Include the extraction of following parts by weight
Object:2 parts of date polysaccharide extract, 1 part of Lycium barbarum polysaccharide extract, 2 parts of tremella polysaccharides extract, neutral notoginseng polysaccharide extract 2
Part.
4. jujube complex polysaccharide composition according to claim 1, it is characterised in that：It further include pharmaceutic adjuvant.
5. jujube complex polysaccharide composition according to claim 1, it is characterised in that：Its dosage form is granule, tablet, glue
Any one in wafer or oral solution.
6. the preparation method of the jujube complex polysaccharide composition according to any one of claim 1-4 claims,
It is characterized in that：Include the following steps：
（1）The extraction of date polysaccharide；
（2）The extraction of polysaccharides；
（3）The extraction of tremella polysaccharides；
（4）The extraction of neutral notoginseng polysaccharide；
（5）Said extracted object, packing are mixed according to weight proportion.
7. the preparation method of jujube complex polysaccharide composition according to claim 6, it is characterised in that：The step（1）
The extraction specific method of middle date polysaccharide is：Jujube crushes, and more times of amount water are added and impregnate, multi-reflow extraction discards filter residue conjunction
And filtrate, concentration are added ethyl alcohol alcohol precipitation, pump supernatant and precipitated and dried, obtain dried powder.
8. the preparation method of jujube complex polysaccharide composition according to claim 7, it is characterised in that：The tool of the alcohol precipitation
Body method is：The filtrate of separating and filtering is concentrated into relative density 1.06, and 3 times of 95% ethyl alcohol is added to alcohol content 75%, alcohol precipitation time
For for 24 hours.
9. the preparation method of jujube complex polysaccharide composition according to claim 7, it is characterised in that：The multi-reflow
The specific method of extraction is：It is added the 8-10 times of water measured, first refluxing extraction 5-8h filters out solution, adds 5-10 times and measures
Secondary water refluxing extraction 2-4h, filters out solution, adds the 4-6 times of refluxing extraction 2-4h of water three times measured, discards filter residue and merges filter
Liquid.
10. the preparation method of jujube complex polysaccharide composition according to claim 6, it is characterised in that：The step（2）
The extraction specific method of middle polysaccharides is：Matrimony vine is added more times of amount water and impregnates, and multi-reflow extraction, merging filtrate concentrates, point
From filtering solution, ethyl alcohol alcohol precipitation is added after concentration, pumps supernatant and is precipitated and dried, obtain dried powder.
11. the preparation method of jujube complex polysaccharide composition according to claim 10, it is characterised in that：It is described repeatedly to return
Flowing the specific method extracted is：First 9-11 times of addition is measured water and is impregnated 12 hours, and first refluxing extraction 5-8h filters out solution, then
The 4-10 times of water refluxing extraction 2-4h measured is added, it is secondary to filter out solution, water that 3-5 times is measured refluxing extraction 2- three times is added later
4h discards filter residue merging filtrate.
12. the preparation method of jujube complex polysaccharide composition according to claim 6, it is characterised in that：The step（3）
The extraction specific method of middle tremella polysaccharides is：White fungus is added more times of amount water and impregnates, multi-reflow extraction, merging filtrate, and concentration adds
Enter ethyl alcohol alcohol precipitation, pumps supernatant and precipitated and dried, obtain dried powder.
13. the preparation method of jujube complex polysaccharide composition according to claim 12, it is characterised in that：It is described repeatedly to return
Flowing the specific method extracted is：First 48-52 times of addition is measured water and is impregnated 12 hours, and first refluxing extraction 5-8h filters out solution, then
The 30-40 times of water refluxing extraction 3-5h measured is added, it is secondary to filter out solution, merging filtrate.
14. the preparation method of jujube complex polysaccharide composition according to claim 6, it is characterised in that：The step（4）
Neutral notoginseng polysaccharide is the waste residue by extracting arasaponin, through water extract-alcohol precipitation, DEAE Sepharose Fast Flow
Anion-exchange chromatography after purification gained.
15. jujube complex polysaccharide composition according to any one of claim 1 to 5 is preparing raising immunity, improve
Application in apathetic, physically weak weak, appetite reduction, nausea and vomiting, loose stool diarrhea, irritated insomnia drug.
16. jujube complex polysaccharide composition according to any one of claim 1 to 5 is preparing raising immunity, improve
Application in apathetic, physically weak weak, appetite reduction, nausea and vomiting, loose stool diarrhea, irritated insomnia health food.
17. jujube complex polysaccharide composition according to any one of claim 1 to 5 is preparing treatment malignant tumor chemotherapy
The application of Neuroleptic Leukocytopenia pharmaceutical preparations afterwards.
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