CN106822228B - Subprostrate sophora polysaccharide effective part and preparation method thereof - Google Patents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/489—Sophora, e.g. necklacepod or mamani
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
Abstract
The invention discloses a subprostrate sophora polysaccharide effective part and a preparation method thereof, wherein the subprostrate sophora polysaccharide effective part is obtained by carrying out alcohol precipitation on water extract of subprostrate sophora and then carrying out fractional drying, wherein the mass percent of total polysaccharide is more than or equal to 65%, and the mass percent of matrine is less than 0.5%. Experiments show that: the acute toxicity and the liver toxicity of the subprostrate sophora polysaccharide effective part provided by the invention are both obviously reduced, and the subprostrate sophora polysaccharide has obvious immunity enhancement effect, and has important significance for researching and developing subprostrate sophora pharmaceutical preparations with clinical application value; and the preparation method is simple, good in repeatability, high in yield, suitable for large-scale production and high in industrial application value.
Description
Technical Field
The invention relates to a subprostrate sophora polysaccharide effective part and a preparation method thereof, belonging to the technical field of medicines.
Background
The radix Sophorae Tonkinensis is dried root and rhizome of Sophora tonkinensis (Sophora tonkinensis Gagnep) of Leguminosae, is mainly produced in Guangxi, Guangdong, Guizhou provinces, has bitter and cold nature, and enters lung meridian. The pharmacopoeia records that the subprostrate sophora has the efficacies of clearing away heat and toxic material, relieving sore throat and dissipating swelling and pain. The subprostrate sophora comprises the following main components: alkaloids, flavonoids, organic acids, saponins and polysaccharides. Pharmacological experiments show that the subprostrate sophora has various pharmacological activities such as anti-tumor, anti-virus, anti-hepatitis, anti-arrhythmia and the like, and the alkaloid components contained in the subprostrate sophora are the material basis of the pharmacological activities according to research reports. Modern researches show that the subprostrate sophora has obvious curative effect on hepatitis, for example, long-term clinical application of a hepatitis treating injection containing subprostrate sophora total alkaloids shows that the injection can reduce transaminase, improve immunity of organisms and has better curative effect on chronic active hepatitis. For example, chinese patent CN85100392A discloses a method for extracting a drug for treating hepatitis from a plant subprostrate sophora, and chinese patent CN1457778A discloses an oral preparation of subprostrate sophora, a preparation method and an application thereof. All the patents refer to a alkaloid component which is a main secondary metabolite in the subprostrate sophora.
However, animal experimental studies show that: the subprostrate sophora has liver toxicity, obvious liver injury can be caused by giving 16g/kg of subprostrate sophora water decoction to rats, the injury mechanism of the subprostrate sophora water decoction is related to the action of inflammatory factors and lipid peroxidation, and the subprostrate sophora water decoction has certain similarity with the liver toxicity of carbon tetrachloride (China journal of Experimental prescriptions, Vol. 19, No. 18, 2013, No. 9, p. 293-. And preclinical studies found: the total extract (mainly alkaloid and flavonoid) of the subprostrate sophora which is soluble in water and ethanol is perfused into the stomach (25g/kg) of a mouse, and symptoms such as respiratory depression, ultra-large tremor, convulsion and the like appear, even the death of the animal is caused. Therefore, studies on the activity of non-alkaloid components of subprostrate sophora have recently been conducted, such as: chinese patent CN1306854A proposes a water extraction and alcohol precipitation part of subprostrate sophora, and the specific preparation method is: soaking radix Sophorae Tonkinensis in water, heating for extraction, extracting with hot water for 0-3 times, filtering, mixing water extractive solutions, concentrating, adding ethanol, and precipitating with ethanol. "the patent indicates that the water and alcohol precipitate in the subprostrate sophora is mainly the non-alkaloid part of macromolecular substances such as polysaccharide; in addition, the experiments of this patent show that: the water and alcohol precipitation in the subprostrate sophora can reduce the CCL4The acute liver injury can reduce the death rate of rats from 30% to 10%, and has liver protecting and immunoregulation effects. However, no research report about acute toxicity and liver toxicity of the subprostrate sophora polysaccharide effective part and the research report about the medicinal value of the part after toxicity is reduced are found so far.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to provide the subprostrate sophora polysaccharide effective part with low acute toxicity and liver toxicity and remarkable immunity enhancing activity and the preparation method thereof, so as to widen the clinical application of the subprostrate sophora.
The subprostrate sophora polysaccharide effective part is obtained by carrying out alcohol precipitation on water extract of subprostrate sophora and then drying by stages, wherein the mass percent of the total polysaccharide is more than or equal to 65 percent, and the mass percent of the matrine is less than 0.5 percent.
Preferably, the mass percent of the total polysaccharide contained in the subprostrate sophora polysaccharide effective part is more than or equal to 70%.
As a further preferable scheme, the mass percent of the total polysaccharide contained in the subprostrate sophora polysaccharide effective part is more than or equal to 85 percent.
The preparation method of the subprostrate sophora polysaccharide effective part comprises the following specific steps:
a) decocting radix Sophorae Tonkinensis with water to obtain radix Sophorae Tonkinensis water decoction;
b) concentrating the subprostrate sophora water decoction at 75-80 ℃ under reduced pressure until the concentration is 1-2 kg/L, then heating at 95-100 ℃ for 1-2 hours under normal pressure, and cooling to room temperature to obtain the subprostrate sophora water extract;
c) adding 90-100% by volume of ethanol into the water extract of the subprostrate sophora obtained in the step b), stirring while adding until the final volume fraction of the ethanol is 75-85%, standing, and collecting precipitates;
d) and (3) carrying out fractional reduced pressure drying on the obtained precipitate: drying under reduced pressure at 80 +/-2 ℃ for 10-14 hours, and then drying under reduced pressure at 75 +/-2 ℃ until the subprostrate sophora polysaccharide is completely dried, thus obtaining the subprostrate sophora polysaccharide effective part.
As a preferred scheme, the preparation of the subprostrate sophora water decoction comprises the following operations:
soaking the medicinal material of the subprostrate sophora in water, heating to 95-100 ℃, and decocting for 1-3 times, wherein the decocting time is 0.5-1.5 hours each time.
As a preferred scheme, the weight of the water added in each decoction is 4-8 times of that of the subprostrate sophora.
Compared with the prior art, the invention has the following remarkable beneficial effects:
1. through test analysis: the subprostrate sophora polysaccharide effective part provided by the invention contains more than or equal to 65% of total polysaccharide by mass and less than 0.5% of matrine by mass; compared with the existing water extraction and alcohol precipitation part, the acute toxicity and the liver toxicity of the subprostrate sophora polysaccharide effective part provided by the invention are remarkably reduced, which shows that the safety of the subprostrate sophora polysaccharide effective part is remarkably improved by controlling the content of matrine;
2. in vitro and in vivo experiments show that: the subprostrate sophora polysaccharide effective part provided by the invention has obvious immunity enhancement effect, which shows that the toxicity is obviously reduced and the subprostrate sophora polysaccharide has obvious immunity enhancement activity by reasonably controlling the contents of total polysaccharide and matrine in the subprostrate sophora polysaccharide effective part, and compared with the prior art, the subprostrate sophora polysaccharide effective part has obvious progress and unexpected effect, and has important significance for researching and developing subprostrate sophora pharmaceutical preparations with clinical application value;
3. in addition, the preparation method is simple, has good repeatability (the similarity of effective part fingerprints prepared in different batches is up to 99 percent), has the yield up to 9.11 percent, is suitable for large-scale production, and has industrial application value.
Drawings
FIG. 1 is a fingerprint of different batches of samples prepared in example 2.
Detailed Description
The present invention will be further illustrated below with reference to specific examples and comparative examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out according to conventional conditions or according to conditions recommended by the manufacturers.
Example 1: preparation of subprostrate sophora polysaccharide effective part
a) Soaking 8kg of radix Sophorae Tonkinensis decoction pieces in 48kg of water for 60min, heating, refluxing and decocting for 1h, filtering the decoction with 200 meshes, adding 48kg of water into the residue, heating, refluxing and decocting for 1h, filtering the decoction with 200 meshes, and mixing the two filtrates to obtain radix Sophorae Tonkinensis water decoction;
b) concentrating the subprostrate sophora water decoction under reduced pressure at 75-80 ℃ until the concentration is 1.5kg/L, then heating at 98 ℃ under normal pressure for 1.5 hours, and cooling to room temperature to obtain the subprostrate sophora water extract;
c) adding ethanol with the volume fraction of 95% into the subprostrate sophora water extract obtained in the step b), stirring while adding until the volume fraction of the final ethanol is 80%, standing, and collecting precipitate;
d) and (3) carrying out fractional reduced pressure drying on the obtained precipitate: drying under reduced pressure at 80 deg.C for 12 hr, and drying under reduced pressure at 75 deg.C to dry completely to obtain 728.8g of the effective fraction of radix Sophorae Tonkinensis polysaccharide with yield of 9.11%.
The effective part of the subprostrate sophora polysaccharide contains 88 wt% of sugar by measuring with a phenol-sulfuric acid method and a spectrophotometry, and the sugar is calculated by glucose.
Analysis by HPLC: the effective fraction of radix Sophorae Tonkinensis polysaccharide contains Matrine (MT) 0.46 wt% and Oxymatrine (OMT) 0.046 wt%.
Example 2: stability study of the preparation Process
According to the preparation process of the example 1, three batches of subprostrate sophora polysaccharide effective parts are prepared repeatedly, the fingerprint of the traditional Chinese medicine is prepared by HPLC, and the similarity among different batches of samples is compared. The HPLC conditions are as follows:
the experimental results are shown in fig. 1, wherein R is the sample of example 1, S1, S2, S3 are the samples of batches 1-3 prepared in this example, respectively, as can be seen from fig. 1: the similarity of the fingerprint spectrums of the samples R, S1, S2 and S3 reaches 99 percent, which shows that the preparation process is stable, has good repeatability and is expected to be applied to industrial mass production.
Comparative example: preparation of water extraction and alcohol precipitation part of subprostrate sophora
Refer to chinese patent CN1306854A example 1:
weighing 8kg of chopped subprostrate sophora, soaking in 12 times of water for overnight, heating and extracting for 2h, pouring out water solution, centrifuging when the solution is hot (3000r/min, l0min), continuously adding 10 times of water and 8 times of water into the medicine residue, respectively heating and extracting for 1.5h and 1h, respectively pouring out the water solution, centrifuging when the solution is hot, and concentrating the water solution to 1 in an inverse gradient manner: 1(1g crude drug is equivalent to 1mL), adding ethanol with the volume fraction of 95% to ensure that the alcohol content reaches 80%, standing at low temperature overnight, centrifuging, adding 1: dissolving with 50% hot water, centrifuging, adding 95% ethanol to the aqueous solution to make the ethanol content reach 80%, standing at low temperature overnight, centrifuging, adding 95% ethanol into the precipitate, stirring, centrifuging, vacuum drying the precipitate (40 deg.C), to obtain herba Desmodii Multifloi water extraction and ethanol precipitation part 257.6g, with a yield of 3.20%.
The sugar content of the subprostrate sophora water extraction and alcohol precipitation part is 58 wt% in terms of glucose by adopting a phenol-sulfuric acid method and being measured by a spectrophotometry.
Analysis by HPLC: the water extraction and alcohol precipitation part of the subprostrate sophora contains 0.65 wt% of Matrine (MT) and 0.132 wt% of Oxymatrine (OMT).
Toxicity test
The subprostrate sophora polysaccharide effective part prepared in example 1 and the subprostrate sophora water extraction and alcohol precipitation part prepared in the comparative example are respectively acted on wild AB line zebra fish processed to 5dpf (5 days after fertilization), and the acute toxicity and the liver toxicity are evaluated.
Treating the wild AB line zebra fish juvenile fish at a certain stage to 5dpf with the drug to be tested, setting a series of concentrations, and simultaneously setting a blank control group, wherein 30 zebra fish are treated at each concentration. During the drug treatment period, the death number of the zebra fish of each experimental group is counted every day and removed in time, and LC can be obtained by fitting concentration-death curve through originPro 8.0 software and curve fitting50、LC10The test results are shown in tables 1 to 3.
TABLE 1 mortality of zebrafish induced by the active fraction of subprostrate sophora polysaccharide (n ═ 30)
| Concentration (μ g/mL) | Counting with fish (tail) | Death number (tail) | Mortality (%) |
| 2500 | 30 | 0 | 0 |
| 3000 | 30 | 0 | 0 |
| 3500 | 30 | 1 | 3.3 |
| 4000 | 30 | 2 | 6.7 |
| 4500 | 30 | 5 | 16.7 |
| 5000 | 30 | 30 | 100 |
TABLE 2 mortality of zebrafish induced by water extraction and alcohol precipitation of Sophora tonkinensis (n ═ 30)
| Concentration (μ g/mL) | Counting with fish (tail) | Death number (tail) | Mortality (%) |
| 600 | 30 | 0 | 0 |
| 800 | 30 | 1 | 3.3 |
| 1000 | 30 | 22 | 73.3 |
| 1200 | 30 | 30 | 100 |
| 1400 | 30 | 30 | 100 |
TABLE 3 toxicity comparison
In combination with tables 1 to 3, it can be seen that: the concentration (5000 mug/mL) of the subprostrate sophora polysaccharide effective part required by the induced 100% mortality of the zebra fish is far higher than the concentration (1200 mug/mL) of the subprostrate sophora water extraction and alcohol precipitation part, and in an acute toxicity experiment, LC (liquid chromatography) of the subprostrate sophora polysaccharide effective part10And LC50LC with values of all the subprostrate sophora water extraction and alcohol precipitation parts10And LC50Value 5 times, and LC of the effective part of subprostrata sophora polysaccharide in the liver toxicity test10The value is also LC of the water extraction and alcohol precipitation part of the subprostrate sophora10More than 5 times the value; the invention obviously reduces the toxicity by reasonably controlling the content of the matrine in the subprostrate sophora polysaccharide effective part, obtains significant progress compared with the prior art, and has important significance for researching and developing the subprostrate sophora pharmaceutical preparation with clinical application value.
Second, in vitro test of immunoregulatory Activity on Normal mouse lymphocytes
The immunoregulatory effect of the subprostrate sophora polysaccharide effective part prepared in example 1 and the subprostrate sophora water extraction and alcohol precipitation part prepared in the comparative example on the mitogen ConA-induced mouse T lymphocyte proliferation function was studied by observing the influence.
(1) Preparation of splenic lymphocytes: the BALB/C mouse is killed after the spine is removed, the spleen of the BALB/C mouse is taken out in an aseptic mode to prepare single cell suspension, and after red blood cells are removed, RPMI-1640 culture solution containing 10% FBS is used for adjusting the cell concentration;
(2) lymphocyte proliferation assay: adding lymphocyte suspension 4X 10 into 96-well cell culture plate5Taking 50 mu L ConA (final concentration is 5 mu g/mL), taking the subprostrate sophora polysaccharide effective part and the subprostrate sophora water extraction and alcohol precipitation part, and respectively preparing administration solutions with final concentrations of 100 mu g/mL, 50 mu g/mL and 25 mu g/mL; each concentration is three-time, the total volume of each hole is 200 mu L, and corresponding non-ConA control holes and non-drug control holes are arranged; 37 degree, 5% CO2Culturing for 48 hours; at 8 hours before the end of the incubation, 25. mu.L of the solution was added to each well3H-thymidine nucleotide (10. mu. Ci/mL); continuing culturing until the experiment is finished; the cells were collected on a glass fiber membrane using a cell collector, and the cell-DNA-doped cells were read in a Beta counter (Microbeta Trilux, Perkinelmer) after addition of scintillation fluid3The amount of H-TdR, expressed as cpm value, represents the cell proliferation, and the test results are shown in Table 4.
TABLE 4 comparison of the Effect on ConA-induced proliferation of mouse T lymphocytes
In the table, compared with the control group,＊＊＊p<0.001。
as can be seen from table 4: the subprostrate sophora polysaccharide effective part group has significance in the proliferation effect of T lymphocytes of normal mice induced by ConA compared with a control group and has concentration dependence; the subprostrate sophora polysaccharide effective part of the invention can promote the proliferation reaction of normal mouse T lymphocyte induced by ConA, and has obvious enhancement effect on the cellular immune function of mice.
Third, in vivo test on immunocompromised mice
The immune enhancement effect of the subprostrate sophora polysaccharide effective part prepared in example 1 and the subprostrate sophora water extraction and alcohol precipitation part prepared in the comparative example on the whole animal is researched by observing the adjusting function of the mouse model with low immunity caused by irradiation.
Irradiating BALB/C mouse with X-ray at 18-22g under 3GRADE for three minutes; after irradiation, the cells were randomly divided into 5 groups according to body weight, which were: normal, model, low, medium and high dose groups; two weeks after administration, the suspension extract was made up to concentrations of 25mg/kg, 50mg/kg and 100mg/kg with 0.3% CMC-Na, respectively. The test results are as follows:
(1) physical signs and body weight
Fifteen days after molding, the model mouse shows obvious weight loss, dull hair color, lassitude, less motion and less food; the above symptoms of each mouse in the administered group were relieved and improved as compared with the model group.
Fifteen days after administration, p of the body weight of the model group mice is less than 0.01 compared with that of the control group, which indicates that the model group is significantly different from the normal group, and indicates that the model building is successful. However, in comparison with the model group, p >0.05 was not statistically significant, but the weight was recovered, and the detailed results are shown in Table 5.
TABLE 5 weight effects on immunocompromised mouse models
In the table, the results, compared to the model set,＊＊p<0.01。
as can be seen from table 5: the subprostrate sophora polysaccharide effective part group has the function of recovering the weight of a mouse model with low immunity, and the effect is slightly higher than that of the subprostrate sophora water extraction and alcohol precipitation part group.
(2) Size and morphology of immune organs and Thymus and spleen index
The mouse cervical spine was sacrificed and spleen size was observed and weighed, and the detailed results are shown in table 6.
TABLE 6 Effect on spleen weight in immunocompromised mice
In the table, the results, compared to the model set,＊p<0.05，＊＊p<0.01。
as can be seen from table 6: the spleen of the model group is smaller than that of the control group, and p is less than 0.01, which indicates that the model is successfully made; however, the effective part group and the water extraction and alcohol precipitation part group of the subprostrate sophora have rising trends compared with the model group, and particularly, compared with the model group, the 25mg/kg dosage group of the effective part of the subprostrate sophora polysaccharide has p less than 0.05 and has significant difference, which indicates that the 25mg/kg dosage group of the effective part of the subprostrate sophora polysaccharide has obvious immunity enhancement effect.
Thymus and spleen of mice were collected and the mass of the blood was measured after the blood residue was blotted with filter paper, and divided by the body mass of the mice and multiplied by 10 to obtain thymus index and spleen index, and the detailed results are shown in Table 7.
TABLE 7 Effect on spleen index and thymus index in immunocompromised mice
In the table, the results, compared to the model set,＊p<0.05。
as can be seen from table 7: the thymus and spleen index of the model group is compared with that of the control group, and p is less than 0.05, which indicates that the model of the low immunity model is successfully made; however, the effective part group of the subprostrate sophora polysaccharide and the water extraction and alcohol precipitation part group of the subprostrate sophora can stimulate the thymus mass of the spleen of a mouse to increase after administration, the thymus index of the effective part group of the subprostrate sophora polysaccharide is increased more obviously, and particularly, the thymus index of the effective part group of the subprostrate sophora polysaccharide of 25mg/kg is increased by statistical difference, which shows that the effective part of the subprostrate sophora polysaccharide has obvious immunity enhancement effect on the mouse with low immunity, and the immunity enhancement effect is obviously higher than that of the effective part of the subprostrate sophora water extraction and alcohol precipitation.
Fourth, ConA stimulation of mouse spleen cell proliferation assay
In order to confirm whether the immune enhancing capability of the subprostrate sophora polysaccharide effective part and the subprostrate sophora water extraction and alcohol precipitation part has concentration dependence, a ConA stimulation mouse spleen cell proliferation experiment is further carried out:
one hour after dosing, mice of different groups were sacrificed by cervical dislocation; taking out spleen, grinding spleen, and mouse spleen lymphocyte 4 × 105One/well was seeded in a 96-well plate,simultaneously adding ConA to make the final concentration of the mixture to be 0.5 mu g/mL, wherein each sample has three composite holes, and the total volume of each hole is 200 mu L; 5% CO2Culturing in incubator for 48 hr, and adding 0.25 μ Ci 8 hr before the end of culturing3H-thymidine, and when the culture is finished, freezing the culture plate in a refrigerator at the temperature of-20 ℃; during measurement, the frozen and thawed cells are collected on a glass fiber membrane by using a cell collector, 3H-thymidine doped with cell DNA is read by a Beta counter after scintillation fluid is added, and the cell proliferation condition is represented by a cpm value; the detailed results are shown in Table 8.
TABLE 8 in vivo cell proliferation assay results
In the table, the results, compared to the model set,＊p<0.05，＊＊p<0.01，＊＊＊p<0.001。
as can be seen from table 8: compared with the model group, the two administration groups of the subprostrate sophora polysaccharide effective part group and the subprostrate sophora water extraction and alcohol precipitation part group can improve the proliferation capacity of immune cells in the spleen, and have concentration dependence.
In combination with tables 4 to 8, it can be seen that: the subprostrate sophora polysaccharide effective part has good immunity enhancement effect, and the activity of the subprostrate sophora polysaccharide effective part is superior to that of the subprostrate sophora water extraction and alcohol precipitation part.
In conclusion, the experiment shows that: the invention not only obviously reduces the toxicity, but also has obvious immunity enhancing activity by controlling the contents of the total polysaccharide and the matrine in the subprostrate sophora polysaccharide effective part, obtains obvious progress and unexpected effect compared with the prior art, and has important significance for researching and developing the subprostrate sophora pharmaceutical preparation with clinical application value; moreover, the preparation method is simple, high in yield, good in stability, easy to realize large-scale production and has the effect of promoting wide application of the subprostrate sophora.
Finally, it should be pointed out here that: the above is only a part of the preferred embodiments of the present invention and should not be construed as limiting the scope of the present invention, and the insubstantial modifications and adaptations of the present invention by those skilled in the art based on the above description are intended to be covered by the present invention.
Claims (1)
1. A method for preparing the subprostrate sophora polysaccharide effective part is characterized by comprising the following specific steps:
a) soaking 8Kg of radix Sophorae Tonkinensis decoction pieces in 48Kg of water for 60min, heating, refluxing and decocting for 1h, filtering the decoction with 200 mesh, adding 48Kg of water into the residue, heating, refluxing and decocting for 1h, filtering the decoction with 200 mesh, and mixing the two filtrates to obtain radix Sophorae Tonkinensis water decoction;
b) concentrating the subprostrate sophora water decoction under reduced pressure at 75-80 ℃ until the concentration is 1.5kg/L, then heating at 98 ℃ under normal pressure for 1.5 hours, and cooling to room temperature to obtain the subprostrate sophora water extract;
c) adding ethanol with the volume fraction of 95% into the subprostrate sophora water extract obtained in the step b), stirring while adding until the volume fraction of the final ethanol is 80%, standing, and collecting precipitate;
d) and (3) carrying out fractional reduced pressure drying on the obtained precipitate: drying under reduced pressure at 80 deg.C for 12 hr, and drying under reduced pressure at 75 deg.C to dry completely, wherein the effective fraction of radix Sophorae Tonkinensis polysaccharide contains total polysaccharide more than or equal to 65 wt%, and matrine less than 0.5 wt%.
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