CN113713074A - Poly-gamma-glutamic acid/chitosan micro-capsule decoction pieces embedded with anti-inflammatory and analgesic components and preparation method thereof - Google Patents
Poly-gamma-glutamic acid/chitosan micro-capsule decoction pieces embedded with anti-inflammatory and analgesic components and preparation method thereof Download PDFInfo
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- CN113713074A CN113713074A CN202111042836.4A CN202111042836A CN113713074A CN 113713074 A CN113713074 A CN 113713074A CN 202111042836 A CN202111042836 A CN 202111042836A CN 113713074 A CN113713074 A CN 113713074A
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Abstract
The invention belongs to the technical field of medicine preparation, relates to a novel composite carrier material, and particularly relates to a poly-gamma-glutamic acid/chitosan microcapsule decoction piece embedded with an anti-inflammatory and analgesic component and a preparation method thereof. The poly-gamma-glutamic acid/chitosan microcapsule decoction piece embedded with the anti-inflammatory and analgesic components is characterized in that the microcapsule decoction piece is prepared by taking poly-gamma-glutamic acid/chitosan as a carrier and loading a traditional Chinese medicine extract. The traditional Chinese medicine extract comprises the following raw materials: turmeric, rosemary, Brazilian pepper, hawthorn, poria cocos, acanthopanax, lucid ganoderma, tripterygium wilfordii, Chinese date, loquat leaves, cinnamon, red pepper, vitamin D, golden chanterelle mushroom, liquorice, white quinoa alcohol, mulberry leaves and moringa oleifera. The gamma-PGA-CS microcapsule decoction pieces prepared by the invention have good relieving effect on the embedded traditional Chinese medicine components and high drug loading, can embed various medicines simultaneously, has very high bioavailability, and can reduce side effects to the maximum extent to realize the effects of diminishing inflammation, detoxifying, reducing internal heat, relieving pain and the like.
Description
Technical Field
The invention belongs to the technical field of medicine preparation, relates to a novel composite carrier material, and particularly relates to a poly-gamma-glutamic acid/chitosan microcapsule decoction piece embedded with an anti-inflammatory and analgesic component and a preparation method thereof.
Background
With the continuous development of life, the pressure caused by the career, family and interpersonal relationship in life is becoming a common phenomenon, and the pressure causes great burden to the human body, which causes the inflammation, fire and qi or pain of a certain part of the body. As is well known, the problems are solved only by pain relievers in western medicines for a long time, so that the problems are solved by traditional Chinese medicines in more cases. However, some traditional Chinese medicines are difficult to absorb and utilize by a human body due to the insolubility, so that the bioavailability of the medicines is greatly reduced. Therefore, improving the bioavailability of some poorly soluble drugs is an important content of the pharmacy of traditional Chinese medicine.
The invention mixes some common traditional Chinese medicines with the effects of diminishing inflammation, reducing internal heat and relieving pain and almost having no side effect according to a proper proportion and prepares the traditional Chinese medicine mixture powder, and the selected traditional Chinese medicines comprise: curcuma longa, Rosemary, Brazil pepper, Crataegus pinnatifida, Poria cocos, Acanthopanax senticosus, Ganoderma lucidum, Tripterygium wilfordii, Zizyphi fructus and Eriobotrya japonica leaves, Cinnamomum cassia, Capsicum annuum, vitamin D, Collybia gallica, Glycyrrhiza uralensis, resveratrol (grape, blueberry), Morus alba, Morus oleifera and the like. Wherein the turmeric is an effective anti-inflammatory agent, and can be used for treating rheumatoid arthritis; brazil pepper can also be used for antibiosis; tripterygium wilfordii also has anti-inflammatory and toxic materials clearing away effects; the golden pheasant mushroom also has anti-inflammatory and antioxidant properties; moringa oleifera also contains various anti-inflammatory and antioxidant substances. In addition, some other medicines also have the auxiliary effects of relieving pain, removing blood stasis, tonifying qi, soothing the nerves and the like. The multiple traditional Chinese medicines are mixed in a certain proportion, so that the mutual drug effects are fully exerted. The efficacy value of the traditional Chinese medicines is fully verified, but the utilization rate of the traditional Chinese medicines is low, so that the problem of further improving the biological activity and the utilization rate of the traditional Chinese medicines is urgently needed to be solved.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a poly-gamma-glutamic acid/chitosan microcapsule decoction piece embedded with anti-inflammatory and analgesic components, which utilizes poly-gamma-glutamic acid and chitosan as carriers, enhances the biocompatibility of the poly-gamma-glutamic acid and the chitosan, enables the poly-gamma-glutamic acid and the chitosan to be better absorbed by organisms, and has good dispersibility due to the existence of the poly-gamma-glutamic acid.
The invention also provides a preparation method of the poly-gamma-glutamic acid/chitosan microcapsule decoction pieces embedded with the anti-inflammatory and analgesic components.
In order to achieve the purpose, the invention relates to the following specific technical scheme:
a poly-gamma-glutamic acid/chitosan microcapsule decoction piece embedded with anti-inflammatory and analgesic components is prepared by taking poly-gamma-glutamic acid/chitosan as a carrier and loading a traditional Chinese medicine extract;
the traditional Chinese medicine extract is prepared by extracting the following raw materials in percentage by mass: 20% of turmeric, 3% of rosemary, 2% of Brazilian pepper, 5% of hawthorn, 7% of poria cocos, 4% of acanthopanax, 8% of lucid ganoderma, 5% of tripterygium wilfordii, 2% of jujube, 1% of loquat leaf, 2% of cinnamon, 1% of red pepper, 5% of vitamin D, 11% of golden chicken oil mushroom, 1% of liquorice, 12% of white quinoa alcohol, 1% of mulberry leaf and 10% of moringa oleifera.
The invention also provides a preparation method of the poly-gamma-glutamic acid/chitosan microcapsule decoction piece, which comprises the following steps:
(1) mixing the raw materials in proportion and grinding into powder to obtain mixed powder, dissolving the mixed powder in anhydrous ethanol to obtain solution B;
(2) adding poly-gamma-glutamic acid into an adipic acid aqueous solution to obtain a poly-gamma-glutamic acid solution C, adding the solution B into the solution C, stirring, and performing rotary evaporation to obtain a mixed solution D;
(3) dissolving chitosan in an acetic acid solution to obtain a mixed solution E, adding cross-linking agents NHS and EDC into the mixed solution D, uniformly stirring, performing ultrasonic treatment to obtain a solution F, adding the solution E into the solution F, continuously stirring in the process of adding the solution E, and performing ultrasonic treatment after the addition is finished to obtain a final solution G;
(4) adjusting the pH value of the solution G, carrying out shaking table reaction, centrifuging, cleaning, freeze-drying to obtain powder, and making into microcapsule decoction pieces by a tablet press.
Further, in the step (1), the concentration of the solution B is 2-5 mg/mL.
Further, in the step (2), the concentration of the poly gamma-glutamic acid solution C is 1 mg/mL; the mass concentration of the adipic acid aqueous solution is 3.5-4%; the volume ratio of the solution B to the poly gamma-glutamic acid solution C is 3: 1.
Further, in the step (2), the stirring is carried out at a rotation speed of 1000-; the rotary evaporation is carried out by using a rotary evaporator, and the rotary evaporation temperature is 40-50 ℃ under the condition of 150-175 mbar.
Further, in the step (3), the concentration of the mixed solution E is 1 mg/mL; the concentration of the glacial acetic acid solution is 2% -7%; the mass ratio of the EDC to the poly gamma-glutamic acid is 2: 1; the mass ratio of the NHS to the poly gamma-glutamic acid is 3: 1; the volume ratio of the solution E to the solution F is as follows: 2:1.
Further, in the step (3), the ultrasound is performed for 10min at a power of 150-.
Further, in the step (4), the pH value is 7-7.4; the shaking table reaction is carried out at the temperature of 37 ℃ and the speed of 180r/min for 2-5 h; the centrifugation is 12000r/min and 10 min.
The invention utilizes gamma-polyglutamic acid (Poly-gamma-glutamic acid, (gamma-PGA)) and chitosan to construct the composite drug carrier. The poly-gamma-glutamic acid is a typical polyelectrolyte, is an amino polymer formed by polymerizing D-glutamic acid and L-glutamic acid through gamma-glutamine bond, has the relative molecular weight of 10-100 ten thousand generally, has excellent biocompatibility, low immunogenicity and no toxic or side effect, and has great advantage as a drug carrier. Chitosan (Chitosan, (CS)) is a functional oligosaccharide, which can reduce cholesterol, enhance immunity, enhance disease resistance and infection resistance of the body, and has excellent biocompatibility.
The invention has the beneficial effects that:
(1) the poly-gamma-glutamic acid and the chitosan used in the invention are colorless, nontoxic, tasteless and easily degradable microbial fermentation extracts, the carboxyl of the gamma-PGA and the amino of the CS have high coordination coefficient and stable structure, the stability of the medicine in the poly-gamma-PGA and the poly-gamma-PGA have good slow release effect and biocompatibility, and the bioavailability of the medicine is higher.
(2) The gamma-PGA-CS microcapsule decoction pieces prepared by the invention have good relieving effect on the embedded traditional Chinese medicine components and high drug loading, can embed various medicines simultaneously, has very high bioavailability, and can reduce side effects to the maximum extent to realize the effects of diminishing inflammation, detoxifying, reducing internal heat, relieving pain and the like.
(3) The gamma-PGA-CS microcapsule decoction piece prepared by the invention has simple preparation method and lower raw material cost.
Drawings
FIG. 1 is a standard curve of drug loading.
Detailed Description
The present invention will be described in further detail with reference to specific embodiments, but it should not be construed that the scope of the present invention is limited to the following examples. Various substitutions and alterations can be made by those skilled in the art and by conventional means without departing from the spirit of the method of the invention described above.
Example 1
(1) Mixing the Chinese medicinal materials at a certain ratio, and grinding into powder to obtain mixed powder with a certain ratio shown in Table 1;
TABLE 1
(2) Because of the insolubility of some traditional Chinese medicines, the mixed powder of the traditional Chinese medicines is dissolved in absolute ethyl alcohol to obtain a traditional Chinese medicine solution with the concentration of 2 mg/mL;
(3) adding poly-gamma-glutamic acid into adipic acid aqueous solution with the mass concentration of 3.5% to prepare poly-gamma-glutamic acid solution with the concentration of about 1mg/mL, adding the traditional Chinese medicine solution into the poly-gamma-glutamic acid solution in a volume ratio of 1:3, stirring, evaporating ethanol on a rotary evaporator to obtain a solution, and setting the water temperature of the evaporator to be 40 ℃ to obtain a mixed solution;
(4) dissolving chitosan in 2% acetic acid solution to obtain 1mg/mL chitosan solution, adding cross-linking agents NHS and EDC into the solution obtained in the step (3), uniformly stirring, performing ultrasonic treatment at 150W for 10min, adding the chitosan solution, wherein the volume ratio of the added chitosan solution to the obtained mixed solution is 2:1, continuously stirring in the adding process, and performing ultrasonic treatment after the adding is finished to obtain the final solution;
(5) and (3) adjusting the pH value of the solution obtained in the step (4) to 7.4 by using PBS buffer solution, placing the solution on a shaking table for reaction for 2 hours, centrifuging, re-suspending the PBS, centrifuging again, freeze-drying the precipitate into powder, and then putting the powder into a tablet press to prepare the microcapsule decoction pieces.
Example 2
(1) Mixing the Chinese medicinal materials in proportion and grinding into powder to obtain mixed powder, wherein the proportion is as in example 1:
(2) because of the insolubility of some traditional Chinese medicines, the mixed powder of the traditional Chinese medicines is dissolved in absolute ethyl alcohol to obtain a traditional Chinese medicine solution with the concentration of 3 mg/mL;
(3) adding poly-gamma-glutamic acid into adipic acid aqueous solution with the mass concentration of 3.5% to prepare poly-gamma-glutamic acid solution with the concentration of about 1mg/mL, adding the traditional Chinese medicine solution into the poly-gamma-glutamic acid solution in a volume ratio of 1:3, stirring, evaporating ethanol on a rotary evaporator to obtain a solution, and setting the water temperature of the evaporator to be 45 ℃ to obtain a mixed solution;
(4) dissolving chitosan in an acetic acid solution with the concentration of 5% to obtain a 1mg/mL chitosan solution, then adding cross-linking agents NHS and EDC into the solution obtained in the step (3), uniformly stirring, performing ultrasonic treatment at 150W for 10min, adding the chitosan solution, wherein the volume ratio of the added chitosan solution to the obtained mixed solution is 2:1, continuously stirring in the adding process, and performing ultrasonic treatment after the adding is finished to obtain a final solution;
(5) and (3) adjusting the pH value of the solution obtained in the step (4) to 7.2 by using PBS buffer solution, placing the solution on a shaking bed for reaction for 3 hours, centrifuging, re-suspending the PBS, centrifuging again, freeze-drying the precipitate into powder, and then putting the powder into a tablet press to prepare the microcapsule decoction pieces.
Example 3
(1) Mixing the Chinese medicinal materials in proportion and grinding into powder to obtain mixed powder, wherein the proportion is as in example 1:
(2) because of the insolubility of some traditional Chinese medicines, the mixed powder of the traditional Chinese medicines is dissolved in absolute ethyl alcohol to obtain a traditional Chinese medicine solution with the concentration of 4 mg/mL;
(3) adding poly-gamma-glutamic acid into adipic acid aqueous solution with the mass concentration of 3.5% to prepare poly-gamma-glutamic acid solution with the concentration of about 1mg/mL, adding the traditional Chinese medicine solution into the poly-gamma-glutamic acid solution in a volume ratio of 1:3, stirring, evaporating ethanol on a rotary evaporator to obtain a solution, and setting the water temperature of the evaporator to be 45 ℃ to obtain a mixed solution;
(4) dissolving chitosan in an acetic acid solution with the concentration of 5% to obtain a 1mg/mL chitosan solution, then adding cross-linking agents NHS and EDC into the solution obtained in the step (3), uniformly stirring, performing ultrasonic treatment at 150W for 10min, adding the chitosan solution, wherein the volume ratio of the added chitosan solution to the obtained mixed solution is 2:1, continuously stirring in the adding process, and performing ultrasonic treatment after the adding is finished to obtain a final solution;
(5) and (3) adjusting the pH value of the solution obtained in the step (4) to 7.0 by using PBS buffer solution, placing the solution on a shaking bed for reaction for 4 hours, centrifuging, re-suspending the PBS, centrifuging again, freeze-drying the precipitate into powder, and then putting the powder into a tablet press to prepare the microcapsule decoction pieces.
Example 4
(1) Mixing the Chinese medicinal materials in proportion and grinding into powder to obtain mixed powder, wherein the proportion is as in example 1:
(2) because of the insolubility of some traditional Chinese medicines, the mixed powder of the traditional Chinese medicines is dissolved in absolute ethyl alcohol to obtain a traditional Chinese medicine solution with the concentration of 5 mg/mL;
(3) adding poly-gamma-glutamic acid into adipic acid aqueous solution with the mass concentration of 3.5% to prepare poly-gamma-glutamic acid solution with the concentration of about 1mg/mL, adding the traditional Chinese medicine solution into the poly-gamma-glutamic acid solution in a volume ratio of 1:3, stirring, evaporating ethanol on a rotary evaporator to obtain a solution, and setting the water temperature of the evaporator to be 45 ℃ to obtain a mixed solution;
(4) dissolving chitosan in an acetic acid solution with the concentration of 7% to obtain a 1mg/mL chitosan solution, then adding cross-linking agents NHS and EDC into the solution obtained in the step (3), uniformly stirring, performing ultrasonic treatment at 150W for 10min, adding the chitosan solution, wherein the volume ratio of the added chitosan solution to the obtained mixed solution is 2:1, continuously stirring in the adding process, and performing ultrasonic treatment after the adding is finished to obtain a final solution;
(5) and (3) adjusting the pH value of the solution obtained in the step (4) to 7.2 by using PBS buffer solution, placing the solution on a shaking table for reaction for 5 hours, centrifuging, re-suspending the PBS, centrifuging again, freeze-drying the precipitate into powder, and then putting the powder into a tablet press to prepare the microcapsule decoction pieces.
Example 5
(1) Mixing the Chinese medicinal materials in proportion and grinding into powder to obtain mixed powder, wherein the proportion is as in example 1:
(2) because of the insolubility of some traditional Chinese medicines, the mixed powder of the traditional Chinese medicines is dissolved in absolute ethyl alcohol to obtain a traditional Chinese medicine solution with the concentration of 3.5 mg/mL;
(3) adding poly-gamma-glutamic acid into adipic acid aqueous solution with the mass concentration of 3.5% to prepare poly-gamma-glutamic acid solution with the concentration of about 1mg/mL, adding the traditional Chinese medicine solution into the poly-gamma-glutamic acid solution in a volume ratio of 1:3, stirring, evaporating ethanol on a rotary evaporator to obtain a solution, and setting the water temperature of the evaporator to be 45 ℃ to obtain a mixed solution;
(4) dissolving chitosan in 2% acetic acid solution to obtain 1mg/mL chitosan solution, adding cross-linking agents NHS and EDC into the solution obtained in the step (3), uniformly stirring, performing ultrasonic treatment at 150W for 10min, adding the chitosan solution, wherein the volume ratio of the added chitosan solution to the obtained mixed solution is 2:1, continuously stirring in the adding process, and performing ultrasonic treatment after the adding is finished to obtain the final solution;
(5) and (3) adjusting the pH value of the solution obtained in the step (4) to 7.2 by using PBS buffer solution, placing the solution on a shaking bed for reaction for 3 hours, centrifuging, re-suspending the PBS, centrifuging again, freeze-drying the precipitate into powder, and then putting the powder into a tablet press to prepare the microcapsule decoction pieces.
Comparative example 1
(1) Mixing the Chinese medicinal materials at a certain ratio and grinding into powder to obtain mixed powder, wherein the ratio is as in example 1;
(2) because of the insolubility of some traditional Chinese medicines, the mixed powder of the traditional Chinese medicines is dissolved in absolute ethyl alcohol to obtain a traditional Chinese medicine solution with the concentration of 2 mg/mL;
(3) adding water into poly-gamma-glutamic acid to prepare poly-gamma-glutamic acid solution with the concentration of about 1mg/mL, adding the traditional Chinese medicine solution into the poly-gamma-glutamic acid solution with the volume ratio of 1:3, stirring, completely evaporating ethanol from the obtained solution on a rotary evaporator, and setting the water temperature of the evaporator to be 40 ℃ to obtain mixed solution;
(4) dissolving chitosan in 2% acetic acid solution to obtain 1mg/mL chitosan solution, adding cross-linking agents NHS and EDC into the solution obtained in the step (3), uniformly stirring, performing ultrasonic treatment, adding the chitosan solution after performing ultrasonic treatment at 150W for 10min, continuously stirring in the adding process, and performing ultrasonic treatment after completing the adding to obtain the final solution;
(5) and (3) adjusting the pH value of the solution obtained in the step (4) to 7.4 by using PBS buffer solution, placing the solution on a shaking table for reaction for 2 hours, centrifuging, re-suspending the PBS, centrifuging again, freeze-drying the precipitate into powder, and then putting the powder into a tablet press to prepare the microcapsule decoction pieces.
By way of example of the gels prepared in the examples and comparative examples, we carried out the following experiments:
1. drug loading rate determination experiment: (1) determination of the standard curve: dissolving the mixed powder in methanol to obtain solutions with concentration of 0, 0.5, 1, 1.5, 2, and 2.5 μ g/mL, and measuring OD at 652 nm.
(2) Detecting the drug loading rate: the microcapsule samples obtained in examples 1 to 5 were placed in a test tube, dissolved in methanol, and then measured for OD at 652nm to calculate the drug loading. The standard curves and the experimental results are shown in fig. 1 and table 2.
TABLE 2 drug loading
From the experimental data it can be seen that examples 1-5 all have relatively high drug loadings.
2. Drug release detection experiments: the microcapsule decoction piece samples prepared in examples 1-5 were placed in test tubes, kept at 37 ℃ and then added with 25ml of phosphate buffered saline respectively, and the supernatants at 0, 4, 8, 12 and 24h were collected and the concentration of the herbal ingredients in the solution was measured by UV spectrophotometer at each time interval, and the experimental results are shown in Table 3.
TABLE 3
The experimental data show that the drug slow release effect of the carrier is good, and the effects of drug slow release and drug utilization rate improvement are achieved.
3. Rat footpad swelling method: (1) selecting 35 rats, dividing the rats into 7 groups, dividing each group into 5 rats, dividing the rats into 2 control groups and 5 experimental groups, performing intragastric administration on the control group 1 according to 0.8mL of normal saline per 100g of body weight, performing intragastric administration on the control group 2 according to a traditional Chinese medicine solution (the mixed powder prepared in the step (1) in the example 1 is dispersed in the normal saline) of traditional Chinese medicine components of 100mg per 100g of body weight, dissolving the traditional Chinese medicine decoction pieces in the normal saline to form a traditional Chinese medicine solution, and performing intragastric administration according to the proportion of 100mg of the traditional Chinese medicine components per 100g of body weight. The gavage was continued for three days, once a day.
(2) And (3) after the intragastric administration of all rats is finished on the third day, performing inflammation model establishment, extracting 0.1mL of 10% egg white physiological saline solution by using a syringe, injecting a part of solution from the middle part of the right hind foot of the rat subcutaneously upwards, straightening the right hind limb of the rat, and then adjusting a syringe needle to inject the rest solution downwards. The volume change of the hind paw after the rat caused inflammation compared to 0.5h, 1h, 1.5h, 2h, 2.5h before the inflammation, i.e. the swelling degree of the paw (swelling degree = volume of hind paw-volume of forefoot before the inflammation), was observed and recorded. The experimental results are shown in table 4 below.
TABLE 4 swelling changes in the footpads of rats
The experimental result shows that compared with a control group, the traditional Chinese medicine decoction piece can obviously reduce the degree of swelling of the foot of a rat to achieve the aim of diminishing inflammation, so that the traditional Chinese medicine micro-capsule decoction piece is proved to have good potential of diminishing inflammation and relieving pain.
Claims (8)
1. A poly-gamma-glutamic acid/chitosan microcapsule decoction piece embedded with anti-inflammatory and analgesic components is characterized in that the microcapsule decoction piece is prepared by taking poly-gamma-glutamic acid/chitosan as a carrier and loading a traditional Chinese medicine extract;
the traditional Chinese medicine extract is prepared by extracting the following raw materials in percentage by mass: 20% of turmeric, 3% of rosemary, 2% of Brazilian pepper, 5% of hawthorn, 7% of poria cocos, 4% of acanthopanax, 8% of lucid ganoderma, 5% of tripterygium wilfordii, 2% of jujube, 1% of loquat leaf, 2% of cinnamon, 1% of red pepper, 5% of vitamin D, 11% of golden chicken oil mushroom, 1% of liquorice, 12% of white quinoa alcohol, 1% of mulberry leaf and 10% of moringa oleifera.
2. The method for preparing the poly-gamma-glutamic acid/chitosan microcapsule decoction piece as claimed in claim 1, comprising the following steps:
(1) mixing the raw materials in proportion and grinding into powder to obtain mixed powder, dissolving the mixed powder in anhydrous ethanol to obtain solution B;
(2) adding poly-gamma-glutamic acid into an adipic acid aqueous solution to obtain a poly-gamma-glutamic acid solution C, adding the solution B into the solution C, stirring, and performing rotary evaporation to obtain a mixed solution D;
(3) dissolving chitosan in an acetic acid solution to obtain a mixed solution E, adding cross-linking agents NHS and EDC into the mixed solution D, uniformly stirring, performing ultrasonic treatment to obtain a solution F, adding the solution E into the solution F, continuously stirring in the process of adding the solution E, and performing ultrasonic treatment after the addition is finished to obtain a final solution G;
(4) adjusting the pH value of the solution G, carrying out shaking table reaction, centrifuging, cleaning, freeze-drying to obtain powder, and making into microcapsule decoction pieces by a tablet press.
3. The method according to claim 2, wherein in the step (1), the concentration of the solution B is 2 to 5 mg/mL.
4. The production method according to claim 2 or 3, wherein in the step (2), the concentration of the poly γ -glutamic acid solution C is 1 mg/mL; the mass concentration of the adipic acid aqueous solution is 3.5-4%; the volume ratio of the solution B to the poly gamma-glutamic acid solution C is 3: 1.
5. The method as claimed in claim 4, wherein in the step (2), the stirring is performed at a rotation speed of 1000-2000r/min for 10-15 min; the rotary evaporation is carried out by using a rotary evaporator, and the rotary evaporation temperature is 40-50 ℃ under the condition of 150-175 mbar.
6. The method according to claim 2, wherein in the step (3), the concentration of the mixed solution E is 1 mg/mL; the concentration of the glacial acetic acid solution is 2% -7%; the mass ratio of the EDC to the poly gamma-glutamic acid is 2: 1; the mass ratio of the NHS to the poly gamma-glutamic acid is 3: 1; the volume ratio of the solution E to the solution F is as follows: 2:1.
7. The preparation method according to claim 6, wherein in the step (3), the ultrasound is performed at a power of 150-200W for 10 min.
8. The production method according to any one of claims 2 to 7, wherein in the step (4), the pH is 7 to 7.4; the shaking table reaction is carried out at the temperature of 37 ℃ and the speed of 180r/min for 2-5 h; the centrifugation is 12000r/min and 10 min.
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Application publication date: 20211130 |