CN105859906B - With immunocompetent sunset abelmoschus stem or bark leaf polyose acetylation modification product of raising and preparation method thereof - Google Patents
With immunocompetent sunset abelmoschus stem or bark leaf polyose acetylation modification product of raising and preparation method thereof Download PDFInfo
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Abstract
Have the invention discloses one kind and improving immunocompetent sunset abelmoschus stem or bark leaf polyose acetylation modification product and preparation method thereof; acetyl content is 16.83% in sunset abelmoschus stem or bark leaf polyose acetylation modification product; polyoses content is 82.50%, and polysaccharide is 0.004 by molar ratio:1.49:0.008:0.011 mannose, glucose, galactolipin and arabinose composition.The present invention passes through the preferred extraction and separation process of many experiments; Thick many candies are obtained using water extraction and alcohol precipitation method; albumen is taken off again; then it is purified using 52 celluosic resins of DEAE; the high sunset abelmoschus stem or bark leaf polyose of purity is made, then modifies using acetic anhydride method to obtain sunset abelmoschus stem or bark leaf polyose acetylation modification product.The present invention makes full use of discarded sunset abelmoschus root cauline leaf resource, turns waste into wealth, and obtains that immunocompetent sunset abelmoschus stem or bark leaf polyose acetylation modification product can be improved the sustainability application, it can be achieved that natural resources of Chinese medicinal materials, has good economic value.
Description
Technical field
The present invention relates to a kind of plant polyoses, and in particular to one kind, which has, improves immunocompetent sunset abelmoschus stem or bark leaf polyose second
Acylated modified outcome and preparation method thereof.
Background technology
Natural resources of Chinese medicinal materials is the solid foundation for ensureing national health, development of national medicine.Chinese medicine and natural medicinal life in recent years
The production area in goods and materials source is more than 2.40 × 106hm2, root yield is up to 5.40 × 106T, and discarded root system of plant and
The biomass of ground cauline leaf is up to 1.1 × 107~1.6 × 107T is 2~3 times of root yield, causes the serious wasting of resources
And environmental pollution.Therefore, the utility value of medicinal organism resource is improved in natural resources of Chinese medicinal materials industrialization process, especially promotes it
Medical value, sound development and development resource economizing type, environmentally friendly economy for Chinese Medicine Industry have great
Meaning.
Sunset abelmoschus root (Abelmoschus manihot L.Medic) is Malvaceae gumbo platymiscium, first recorded in《Jia Yu sheets
Grass》,《Compendium of Materia Medica》Described in：" it spends smell sweet, cold, sliding, nontoxic, cures mainly urine leaching and expedites the emergence of, controls all malignant sore pus long not
The person of being recovered, what work end was applied is cured, for persons particularly liable to develop skin infection's key medicine " etc..Its root, stem, leaf all have certain medical value.Sunset abelmoschus flower is main
Agents area is dropped or burns its stem and leaf part more in recovery process, causes the very big wave of sunset abelmoschus root cauline leaf resource
Take and the pollution of environment.
Polysaccharide is a kind of complicated polymer substance, has antitumor, antiviral, anti-oxidant, anti-mutation, radioresistance
With the various biologicals effect such as enhancing is immune.But the polysaccharide bioactivity under normal circumstances, naturally extracted is weaker.In natural polysaccharide
When introducing certain ionic group in molecule and there is appropriate degree of substitution, the dissolving of polysaccharide in water can not only be significantly improved
Degree, and the chain conformation of polysaccharide can be made to change, improve bioactivity to make it have certain specific structure.It is more
The acetylation modification of sugar can make polysaccharide branch be stretched and be changed, and cause the hydroxyl of polysaccharide to expose, to increase polysaccharide
Solubility in water can cause the bioactivity of original polysaccharide to substantially change.
Currently, the analysis of physicochemical property and monosaccharide is concentrated mainly on to the research of sunset abelmoschus stem or bark leaf polyose both at home and abroad, it is right
Its active research is less.Therefore, change the structure of sunset abelmoschus stem or bark leaf polyose using acetylation modification technology, improve its physics and chemistry
Matter obtains really active polysaccharide, to the efficient utilization of sunset abelmoschus root cauline leaf resource and the Protection significance weight of ecological environment
Greatly.
Invention content
Goal of the invention：The purpose of the present invention is to solve the deficiencies in the prior art, are original with sunset abelmoschus root cauline leaf waste
Material, sunset abelmoschus root cauline leaf Thick many candies are prepared by preferred method, then acetic anhydride method are used to modify, and obtain having raising immune
Active sunset abelmoschus stem or bark leaf polyose acetylation modification product.Another object of the present invention is to provide sunset abelmoschus stem or bark leaf polyose acetylation
The preparation method of modified outcome and its application.The present invention makes full use of discarded sunset abelmoschus root cauline leaf resource, turns waste into wealth, and height is immune
Active sunset abelmoschus stem or bark leaf polyose acetylation modification product, it can be achieved that natural resources of Chinese medicinal materials sustainability application, have well warp
The Protection significance of Ji value and ecological environment.
Technical solution：In order to achieve the goal above, the technical solution adopted by the present invention is：
A kind of sunset abelmoschus stem or bark leaf polyose acetylation modification product, acetyl group in sunset abelmoschus stem or bark leaf polyose acetylation modification product
Content is 16.83%, polyoses content 82.50%, and polysaccharide is 0.004 by molar ratio:1.49:0.008:0.011 mannose,
Glucose, galactolipin and arabinose composition.
Preferably, the molecular weight of sunset abelmoschus stem or bark leaf polyose and sunset abelmoschus stem or bark leaf polyose acetylation modification product is distinguished
For 760.24kDa and 649.52kDa.
Preferably, the preparation method of above-described sunset abelmoschus stem or bark leaf polyose acetylation modification product, including with
Lower step：
(1) water of the dregs of a decoction 10~30 times of volumes of weight, refluxing extraction is added in the dregs of a decoction after taking sunset abelmoschus root cauline leaf ethyl alcohol to extract
2~3 times, 1~2 hour every time, filtering, merging filtrate was concentrated under reduced pressure；Sevag method removing proteins, centrifugation take supernatant, and nothing is added
Water-ethanol, alcohol precipitation is overnight, filters, takes precipitation, washed successively with absolute ethyl alcohol, acetone, ether, dries, it is thick to obtain sunset abelmoschus root cauline leaf
Polysaccharide；
(2) classification of sunset abelmoschus root cauline leaf Thick many candies
The sunset abelmoschus root cauline leaf Thick many candies that step (1) is prepared are weighed, distillation water dissolution is added, are loaded onto DEAE-52 chromatographies
In column, with 0.0,0.1,0.3,0.5mol/L NaCl solution gradient elutions, and polyoses content is detected using phend-sulphuric acid, point
Different eluting peaks is not collected, is concentrated under reduced pressure, and dialysis obtains sunset abelmoschus stem or bark leaf polyose finally by dialyzate vacuum freeze drying；
(3) acetylation modification of sunset abelmoschus stem or bark leaf polyose
The sunset abelmoschus stem or bark leaf polyose for taking step (2) to be prepared, is modified using acetic anhydride method, the specific steps are：It is accurate
Sunset abelmoschus stem or bark leaf polyose really is weighed, is dissolved in distilled water, is 9.0 with sodium hydroxide tune pH, second is alternately added into solution therebetween
Acid anhydrides and sodium hydroxide control temperature to keep pH value of solution for 8.0~8.5, during reaction at 25~30 DEG C, acetic anhydride are added, instead
It is 7.0 with the pH of hydrochloride adjusted solution after answering, mixed liquor, which is placed in flowing water in bag filter, dialyses 2~3 days, takes dialyzate
It is concentrated under reduced pressure, freeze-drying obtains sunset abelmoschus stem or bark leaf polyose acetylation modification product.
Preferably, the preparation method of above-described sunset abelmoschus stem or bark leaf polyose acetylation modification product, step (3)
Accurately to weigh sunset abelmoschus stem or bark leaf polyose 30mg, it is dissolved in 10mL distilled water, 10min dissolvings is stirred in 60 DEG C, are cooled to room temperature
Afterwards, it is 9.0 with 0.5mol/L sodium hydroxide tune pH value, a certain amount of acetic anhydride and 0.5mol/ is alternately added into solution therebetween
LNaOH, to keep pH value of solution for 8.0~8.5；Temperature is controlled during reaction at 30 DEG C, and 150 μ L acetic anhydrides are then added, react
4h, after reaction, the pH that solution is adjusted with 5mol/L HCl are 7.0, and mixed liquor, which is placed in flowing water in bag filter, dialyses 3 days,
It takes dialyzate to be concentrated under reduced pressure, is lyophilized, obtains sunset abelmoschus stem or bark leaf polyose acetylation modification product.
The present invention shows to have by the sunset abelmoschus stem or bark leaf polyose product that acetylation modification obtains aobvious by experimental study
The effect of the promotion spleen lymphocyte proliferation of work, can be used for preparing the drug or health products for improving immunity.
Sunset abelmoschus stem or bark leaf polyose acetylation modification product of the present invention is preparing the drug for improving immunity or health care
Tablet, capsule can be made in sunset abelmoschus stem or bark leaf polyose acetylation modification product and pharmaceutically acceptable carrier by the application in product
Agent, granule or encapsulated form drug.
The present invention is when being made tablet, by sunset abelmoschus stem or bark leaf polyose acetylation modification product and carrier lactose or cornstarch
It is uniformly mixed, magnesium stearate lubricant is added when needing, be uniformly mixed, tablet is made in then tabletting.
When capsule is made, sunset abelmoschus stem or bark leaf polyose acetylation modification product and carrier lactose or cornstarch are mixed
Uniformly, whole grain, it is then encapsulated that capsule is made.
The present invention is when being made granule, sunset abelmoschus stem or bark leaf polyose acetylation modification product and diluent lactose or corn
Starch is uniformly mixed, whole grain, dry, and granule is made.
Advantageous effect：It is provided by the invention have improve immunocompetent sunset abelmoschus stem or bark leaf polyose acetylation modification product and
The prior art, which is compared, to be had a little：
1, the present invention obtains Thick many candies using water extraction and alcohol precipitation method first by the preferred extraction and separation process of many experiments, then
Then deproteination is purified using DEAE-52 celluosic resins, the high sunset abelmoschus stem or bark leaf polyose of purity is made, then again
It is modified using using acetic anhydride method, then purifies, obtain sunset abelmoschus stem or bark leaf polyose acetylation modification product.
The present invention makes full use of discarded sunset abelmoschus root cauline leaf resource, turns waste into wealth, be prepared can be improved it is immunocompetent
Sunset abelmoschus stem or bark leaf polyose acetylation modification product, it can be achieved that natural resources of Chinese medicinal materials sustainability application, have good economic value
With the Protection significance of ecological environment.
2, the present invention by external mouse spleen lymphocyte proliferation activity experiments have shown that, sunset abelmoschus stem or bark leaf polyose has no bright
Aobvious immunoregulatory activity, but after acetylation modification, there is significant immunoregulatory activity in 50~200 μ g/mL, 50
μ g/mL dosage shows strongest proliferation function, proliferation index 1.217, and the highest proliferation index before modifying is only
1.079, achieve extraordinary unexpected technique effect.
Description of the drawings
Fig. 1 is the infrared spectrogram of sunset abelmoschus root cauline leaf Thick many candies.
Fig. 2 is the infrared spectrogram of sunset abelmoschus stem or bark leaf polyose acetylation modification product.
Fig. 3 is the proliferation of splenocytes block diagram of sunset abelmoschus stem or bark leaf polyose and its acetylation modification product.
Specific implementation mode
According to following embodiments, the present invention may be better understood.However, as it will be easily appreciated by one skilled in the art that real
It applies specific material proportion, process conditions and its result described in example and is merely to illustrate the present invention, without that should will not limit
The present invention described in detail in claims processed.
Embodiment 1
The preparation method of sunset abelmoschus stem or bark leaf polyose acetylation modification product, includes the following steps：
(1) preparation of sunset abelmoschus root cauline leaf Thick many candies
Drying dregs of a decoction 100g of the sunset abelmoschus root cauline leaf after alcohol reflux extracts is taken, the dregs of a decoction 30 times of volumes of weight are added
Water, 100 DEG C of water-bath refluxing extractions 3 times, each 1h.Filtering, merging filtrate are concentrated under reduced pressure.Sevag method removing proteins, centrifugation
(4000r/min, 10min) takes supernatant, the absolute ethyl alcohol of tetraploid product is added, and alcohol precipitation is overnight, filters, and precipitation uses nothing successively
Three times, 50 DEG C of drying are to get sunset abelmoschus root cauline leaf Thick many candies for water-ethanol, acetone, ether washing.
(2) classification of sunset abelmoschus root cauline leaf Thick many candies
The sunset abelmoschus root cauline leaf Thick many candies 3g that step (1) is prepared is weighed, adds appropriate distillation water dissolution, prepares mass concentration
For the polysaccharide solution of 30mg/mL, it is loaded onto in DEAE-52 chromatographic columns.With 0.0,0.1,0.3,0.5mol/LNaCl solution gradients
Elution, flow velocity 1.0mL/min (10min/ pipes) detect polyoses content using phend-sulphuric acid, collect different elutions respectively
Peak is concentrated under reduced pressure, dialysis, and finally by dialyzate vacuum freeze drying, obtaining sunset abelmoschus root cauline leaf Thick many candies, (polyoses content reaches
99%).
(3) acetylation modification of sunset abelmoschus stem or bark leaf polyose
The sunset abelmoschus root cauline leaf Thick many candies that step (2) is prepared are taken, are modified using acetic anhydride method.Accurately weigh yellow another name for Sichuan Province
Certain herbaceous plants with big flowers cauline leaf Thick many candies 30mg, is dissolved in 10mL distilled water, and 10min dissolvings are stirred in 60 DEG C, after being cooled to room temperature, use 0.5mol/L
Sodium hydroxide tune pH is 9.0, a certain amount of acetic anhydride and 0.5mol/LNaOH is alternately added into solution therebetween, to keep solution
PH is 8.0~8.5.Temperature is controlled during reaction at 30 DEG C, 150 μ L acetic anhydrides are added, and reaction 4h uses 5mol/ after reaction
The pH of L HCl adjustment solution is 7.0, and mixed liquor, which is placed in flowing water in bag filter, dialyses 3 days, is concentrated under reduced pressure, freeze-drying obtains sunset abelmoschus root
Stem leaf polysaccharide acetylation modification product.
The measurement of embodiment 2 sunset abelmoschus stem or bark leaf polyose acetylation modification product total sugar content and degree of substitution
1, the measurement of total sugar content and degree of acetylation
Using D- DEXTROSE ANHYDROUSs as standard items, sunset abelmoschus root cauline leaf is thick before and after measuring acetylation modification using phend-sulphuric acid
The total sugar content of polysaccharide；Using determination of acid-basetitration acetyl content, with sodium hydroxide sample dissolution, after abundant saponification, use
The unreacted alkali of titration with hydrochloric acid calculates acetyl content (A%) and degree of substitution (DS) according to the following formula.
In formula：A (%) is acetyl content；V0 (mL) represents the volume of consumed sodium hydroxide solution；C0(mol/L)
Represent the concentration of consumed NaOH solution；V1 (mL) represents the volume of consumed hydrochloric acid solution；C1 (mol/L), which is represented, to be disappeared
The concentration of the hydrochloric acid solution of consumption；M (g) is sample quality.
2, monosaccharide composition analysis
Using trifluoroacetic acid hydrolysis, PMP polysaccharide derivative samples, to mix monosaccharide as standard items, using high performance liquid chromatography
Method determines the composition of monosaccharide according to appearance time, and the monosaccharide that the standard curve determination sample of each monosaccharide is drawn according to peak area contains
Amount.
3, infrared spectrum analysis
Learnt from else's experience dry polysaccharide sample lmg, is ground in agate mortar with the dried potassium bromide powders of 100~200mg
Uniformly, it is pressed into thin slice, 4000~400cm is measured on infrared spectrometer-1Infrared spectrum.
4, mice spleen lymphocytes proliferation is tested
It is sterile to take spleen, mouse spleen lymphocyte suspension is prepared, adjustment cell concentration is 6 × 106A/mL connects in 96 orifice plates
100 holes μ L/ of kind, then culture solution (blank control), 10 μ g/mL ConA (con A, positive control) are separately added into, it is different
The sunset abelmoschus root cauline leaf Thick many candies of concentration and each 100 μ of sunset abelmoschus stem or bark leaf polyose acetylation modification Product samples solution of various concentration
The holes L/, each concentration are all provided with 4 multiple holes, are uniformly mixed and are placed on 37 DEG C, 5%CO2Culture is for 24 hours.Culture terminates preceding 4h, adds per hole
Enter 10 μ L of MTT (5mg/mL), continue to cultivate 4h in above-mentioned condition, supernatant is abandoned in centrifugation, is added 100 μ L DMSO, after fully shaking
The light absorption value of wavelength 570nm is measured with microplate reader.
Lymphopoiesis index=sample sets light absorption value/blank control group light absorption value.
5, experimental result
The measurement analysis result of 5.1 total reducing sugars and degree of substitution, as shown in table 1：
1 sunset abelmoschus root cauline leaf Thick many candies of table and its total sugar content of acetylation modification product and the measurement of degree of substitution
As shown in table 1, the total sugar content of sunset abelmoschus root cauline leaf Thick many candies is 99.76%, not acetyl-containing, the acetyl after modification
Base content is 16.83%, total sugar content 82.50%, the degree of substitution with acetyl group of sunset abelmoschus stem or bark leaf polyose acetylation modification product
It is 0.62, acetyl group has been connected on sugar chain.
The results are shown in Table 2 for 5.2 monosaccharide composition analysis：
The monosaccharide constitutive molar ratio of 2 sunset abelmoschus root cauline leaf Thick many candies of table and its acetylation modification product
The molar ratio of the monosaccharide of sunset abelmoschus root cauline leaf Thick many candies and its acetylation modification product composition is as shown in table 2, sunset abelmoschus root
Cauline leaf Thick many candies are made of mannose, glucose, galactolipin and arabinose, wherein the ratio highest of glucose, Qi Tasan
The ratio of kind sugar is relatively low.After acetylation modification, glucose is mainly contained, mannose is less with galactose content, arabinose
Ratio reduce to less than half original.The molecule of sunset abelmoschus stem or bark leaf polyose and sunset abelmoschus stem or bark leaf polyose acetylation modification product
Amount is respectively 760.24kDa and 649.52kDa.In modification, Partial digestion has occurred in sunset abelmoschus stem or bark leaf polyose.
5.3 infrared spectrum analysis
As depicted in figs. 1 and 2, sunset abelmoschus root cauline leaf Thick many candies are in 3400cm-1The absorption peak of left and right is that the flexible of O-H keys shakes
It is dynamic, 2900cm-1The absorption peak of left and right is the stretching vibration of c h bond, is carbohydrate characteristic peak.And sunset abelmoschus stem or bark leaf polyose acetylation is repaiied
Product is adornd in 1743cm-1There is apparent C=O vibration absorption peaks, 1373cm in place-1Place to be that C-H in acetyl group is flexible shake
Movable property life, and it is located at 1248cm-1What is located is the C-O stretching vibrations in ester group, and the position of other absorption peaks does not change substantially
Become, shows to have had been incorporated into acetyl group on sunset abelmoschus root cauline leaf Thick many candies sugar chain, acetylation modification success.
5.4 proliferation of splenocytes are analyzed
The proliferation of splenocytes experimental result of sunset abelmoschus root cauline leaf Thick many candies and its acetylation modification product is as shown in Figure 3.
A is indicated compared with the control group in figure, humidification significant difference (p ＜ 0.05)；B indicates that compared with the control group, enhancing is made in figure
Extremely significantly (p ＜ 0.01) with difference；C is indicated compared with the control group in figure, and humidification difference is extremely significantly (p ＜ 0.001)；Figure
Middle d is indicated, compared with SLAMP-a groups, humidification significant difference (p ＜ 0.05)；F is indicated in figure, compared with SLAMP-a groups,
Humidification extremely significant difference (p ＜ 0.001).The proliferation index of control group is 1.001, and positive group is 1.509.
From the figure 3, it may be seen that compared with the control group, sunset abelmoschus root cauline leaf Thick many candies cannot pierce in 25~200 μ g/mL concentration ranges
Swash the proliferation of splenic lymphocytes, and its acetylation modification product has the function of promoting spleen lymphocyte proliferation.Sunset abelmoschus root cauline leaf
Polysaccharide acetylation modification product shows strongest proliferation function in 50 μ g/mL dosage, proliferation index 1.217, and before modifying
Highest proliferation index be only 1.079.Achieve extraordinary technique effect.
The technical concepts and features of embodiment of above only to illustrate the invention, its object is to allow be familiar with technique
People understands the content of present invention and is implemented, and it is not intended to limit the scope of the present invention, all according to spirit of that invention reality
The equivalent change or modification that matter is done should all cover within the scope of the present invention.
Claims (4)
1. a kind of solution of sunset abelmoschus stem or bark leaf polyose acetylation modification product, which is characterized in that sunset abelmoschus stem or bark leaf polyose acetylation
Acetyl content is 16.83% in modified outcome, polyoses content 82.50%, and polysaccharide is 0.004 by molar ratio:1.49:
0.008:0.011 mannose, glucose, galactolipin and arabinose composition；Sunset abelmoschus stem or bark leaf polyose acetylation modification produces
A concentration of 50~200 μ g/mL of the solution of object；
The preparation method of hollyhock stem leaf polysaccharide acetylation modification product includes the following steps：
(1) water of the dregs of a decoction 10~30 times of volumes of weight, refluxing extraction 2~3 is added in the dregs of a decoction after taking sunset abelmoschus root cauline leaf ethyl alcohol to extract
Secondary, 1~2 hour every time, filtering, merging filtrate was concentrated under reduced pressure；Sevag method removing proteins, centrifugation, take supernatant, anhydrous second are added
Alcohol, alcohol precipitation is overnight, filters, takes precipitation, washed successively with absolute ethyl alcohol, acetone, ether, dries, obtains sunset abelmoschus root cauline leaf Thick many candies；
(2) classification of sunset abelmoschus root cauline leaf Thick many candies
The sunset abelmoschus root cauline leaf Thick many candies that step (1) is prepared are weighed, adds distillation water dissolution, is loaded onto in DEAE-52 chromatographic columns,
With 0.0,0.1,0.3,0.5mol/L NaCl solution gradient elutions, and polyoses content is detected using phend-sulphuric acid, received respectively
Collect different eluting peaks, be concentrated under reduced pressure, dialysis obtains sunset abelmoschus stem or bark leaf polyose finally by dialyzate vacuum freeze drying；
(3) acetylation modification of sunset abelmoschus stem or bark leaf polyose
The sunset abelmoschus stem or bark leaf polyose for taking step (2) to be prepared, is modified using acetic anhydride method, the specific steps are：It is accurate to claim
Sunset abelmoschus stem or bark leaf polyose is taken, is dissolved in distilled water, is 9.0 with sodium hydroxide tune pH, acetic anhydride is alternately during which added into solution
Temperature is controlled with sodium hydroxide to keep pH value of solution for 8.0~8.5, during reaction at 25~30 DEG C, then adds acetic anhydride,
After reaction, it is 7.0 with the pH of hydrochloride adjusted solution, mixed liquor, which is placed in flowing water in bag filter, dialyses 2~3 days, takes dialysis
Liquid is concentrated under reduced pressure, and freeze-drying obtains sunset abelmoschus stem or bark leaf polyose acetylation modification product.
2. the preparation method of the solution of sunset abelmoschus stem or bark leaf polyose acetylation modification product described in claim 1, which is characterized in that
The preparation method of sunset abelmoschus stem or bark leaf polyose acetylation modification product includes the following steps：
(1) water of the dregs of a decoction 10~30 times of volumes of weight, refluxing extraction 2~3 is added in the dregs of a decoction after taking sunset abelmoschus root cauline leaf ethyl alcohol to extract
Secondary, 1~2 hour every time, filtering, merging filtrate was concentrated under reduced pressure；Sevag method removing proteins, centrifugation, take supernatant, anhydrous second are added
Alcohol, alcohol precipitation is overnight, filters, takes precipitation, washed successively with absolute ethyl alcohol, acetone, ether, dries, obtains sunset abelmoschus root cauline leaf Thick many candies；
(2) classification of sunset abelmoschus root cauline leaf Thick many candies
The sunset abelmoschus root cauline leaf Thick many candies that step (1) is prepared are weighed, adds distillation water dissolution, is loaded onto in DEAE-52 chromatographic columns,
With 0.0,0.1,0.3,0.5mol/L NaCl solution gradient elutions, and polyoses content is detected using phend-sulphuric acid, received respectively
Collect different eluting peaks, be concentrated under reduced pressure, dialysis obtains sunset abelmoschus stem or bark leaf polyose finally by dialyzate vacuum freeze drying；
(3) acetylation modification of sunset abelmoschus stem or bark leaf polyose
The sunset abelmoschus stem or bark leaf polyose for taking step (2) to be prepared, is modified using acetic anhydride method, the specific steps are：It is accurate to claim
Sunset abelmoschus stem or bark leaf polyose is taken, is dissolved in distilled water, is 9.0 with sodium hydroxide tune pH, acetic anhydride is alternately during which added into solution
Temperature is controlled with sodium hydroxide to keep pH value of solution for 8.0~8.5, during reaction at 25~30 DEG C, then adds acetic anhydride,
After reaction, it is 7.0 with the pH of hydrochloride adjusted solution, mixed liquor, which is placed in flowing water in bag filter, dialyses 2~3 days, takes dialysis
Liquid is concentrated under reduced pressure, and freeze-drying obtains sunset abelmoschus stem or bark leaf polyose acetylation modification product.
3. the preparation method of the solution of sunset abelmoschus stem or bark leaf polyose acetylation modification product according to claim 2, feature
It is, step (3) accurately weighs sunset abelmoschus stem or bark leaf polyose 30mg, is dissolved in 10mL distilled water, and 10min dissolvings are stirred in 60 DEG C,
After being cooled to room temperature, it is 9.0 with 0.5mol/L sodium hydroxide tune pH value, a certain amount of acetic anhydride is alternately during which added into solution
With 0.5mol/L NaOH, with keep pH value of solution be 8.0~8.5；Temperature is controlled during reaction at 30 DEG C, and 150 μ L second are then added
Acid anhydrides reacts 4h, and after reaction, the pH that solution is adjusted with 5mol/L HCl is 7.0, and mixed liquor is placed in flowing water in bag filter
Dialysis 3 days, takes dialyzate to be concentrated under reduced pressure, and is lyophilized, obtains sunset abelmoschus stem or bark leaf polyose acetylation modification product.
4. the solution of sunset abelmoschus stem or bark leaf polyose acetylation modification product described in claim 1 improves the drug of immunity preparing
Or the application in health products.
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