CN115282169A - Bufonis venenum extract, and its preparation method and application in preparing medicine for treating heart failure - Google Patents
Bufonis venenum extract, and its preparation method and application in preparing medicine for treating heart failure Download PDFInfo
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Abstract
The invention discloses a preparation method of a toad venom extract, which comprises the following steps: soaking venenum Bufonis powder with softening agent, and standing by after treatment; refluxing in 10-15 times of ethanol solvent, removing ethanol, filtering to obtain first extractive solution, and collecting residue. The venenum bufonis extract is prepared by firstly soaking venenum bufonis powder by using a softening agent, adding chitosan into a sodium alginate solution by using the softening agent, stirring and fully dispersing, and matching modified zeolite powder, wherein the dissolubility of the chitosan is dispersed by using the sodium alginate, and the chitosan is modified by matching the modified zeolite powder, so that the dispersity of the zeolite powder is enhanced after the zeolite powder is modified, and the dispersion of the venenum bufonis powder is promoted in the softening agent, thereby being beneficial to the cooperative extraction in the subsequent reflux extraction and supercritical extraction, and improving the extraction efficiency; the conditioning agent is cooperated with lactose and microcrystalline cellulose, thereby being beneficial to the subsequent application in the medicine for treating heart failure.
Description
Technical Field
The invention relates to the technical field of toad venom extracts, and in particular relates to a toad venom extract, a preparation method thereof and application thereof in preparing a medicine for treating heart failure.
Background
Heart failure is a syndrome in which the heart fails to meet the metabolic demand of tissues due to a decrease in cardiac output caused by primary cardiac damage under normal venous return conditions. Clinically, it is characterized by congestion of the pulmonary and/or systemic circulation and insufficient blood perfusion in the tissues, also called congestive heart failure, which is the terminal stage of heart disease caused by various etiological factors. The concepts of congestive heart failure and cardiac insufficiency are basically the same, but the latter is more broadly defined and includes the stage where there is already a reduction in cardiac output but no clinical symptoms have appeared. The invention provides a toad venom extract, a preparation method thereof and application thereof in preparing a medicine for treating heart failure.
Disclosure of Invention
In view of the defects of the prior art, the present invention aims to provide a toad venom extract, a preparation method thereof and an application thereof in the preparation of a medicament for treating heart failure, so as to solve the problems in the background art.
The technical scheme adopted by the invention for solving the technical problems is as follows:
the invention provides a preparation method of a toad venom extract, which comprises the following steps:
the method comprises the following steps: soaking venenum Bufonis powder with softening agent, and standing by after treatment;
step two: sending into 10-15 times by weight of ethanol solvent for refluxing, removing ethanol after refluxing extraction, filtering to obtain first extractive solution, and collecting residue;
step three: performing supercritical extraction treatment on the filter residue, and obtaining a second extracting solution after extraction is finished;
step four: mixing the first extract and the second extract to obtain an extract, and finally feeding the extract into macroporous resin for elution;
step five: adding 5-10% of conditioning agent into Bufonis venenum extract, stirring, and mixing to obtain Bufonis venenum extract.
Preferably, the preparation method of the softening agent comprises the following steps: adding chitosan into 3-6 times of sodium alginate solution, stirring and dispersing fully, then adding modified zeolite powder accounting for 5-10% of the total amount of chitosan, stirring and mixing fully to obtain a softening agent;
the preparation method of the modified zeolite powder comprises the following steps:
feeding the zeolite powder of 150-200 meshes into a heat treatment at 300-400 ℃ for 20-30min, then cooling to room temperature at the speed of 1-3 ℃/min, then feeding into a hydrochloric acid solution of 3-6 times of mass fraction 5-10% to fully disperse, and washing and drying to obtain the modified zeolite powder.
Preferably, the temperature of the infiltration treatment is 40-45 ℃, the treatment rotating speed is 350-450r/min, and the treatment time is 20-30min.
Preferably, the mass fraction of the sodium alginate solution is 10-15%.
Preferably, the extraction time of the reflux is 40-50min; the mass fraction of the refluxing ethanol solvent is 95-97%.
Preferably, the extraction temperature of the supercritical extraction treatment is 38-42 ℃, the extraction pressure is 28-30MPa, and the extraction time is 1-2h.
Preferably, the specific operation steps of the elution treatment of the macroporous resin are as follows:
and (3) loading the extracting solution on macroporous resin, removing impurities by elution, then eluting, and finally collecting the eluent, wherein the elution is carried out by adopting an ethanol solution with the mass fraction of 55-60%.
The inventor of the invention finds that the preparation of the toad venom extract is not soaked by the softening agent, the extraction rate of the product is obviously reduced, the modified zeolite powder is not added in the preparation of the softening agent and is replaced by zeolite, the extraction rate of the product is in a trend of being poor, meanwhile, the supercritical extraction treatment is not adopted, the extraction rate of the product is further reduced, and the extraction efficiency of the product can be obviously improved by adopting the soaking treatment of the softening agent and the extraction method of the invention.
Preferably, the rotating speed for uniformly stirring and mixing in the step five is 550-650r/min, and the stirring time is 20-30min; the conditioning treatment agent comprises the following raw materials in parts by weight: 5-10 parts of lactose and 1-5 parts of microcrystalline cellulose.
The invention also provides the toad venom extract prepared by the preparation method of the toad venom extract.
The invention also provides application of the toad venom extract in preparing a medicine for treating heart failure.
Compared with the prior art, the invention has the following beneficial effects:
the venenum bufonis extract is prepared by firstly soaking venenum bufonis powder with a molting agent, adding chitosan into a sodium alginate solution to stir and disperse fully, and matching modified zeolite powder, wherein the dissolubility of the chitosan is dispersed through the sodium alginate, and the modified zeolite powder is matched for modification, so that the dispersity of the zeolite powder is enhanced after the zeolite powder is modified, and the dispersion of the venenum bufonis powder is promoted in the molting agent, thereby being beneficial to the cooperative extraction in the subsequent reflux extraction and supercritical extraction, and improving the extraction efficiency; the conditioning agent is cooperated with lactose and microcrystalline cellulose, thereby being beneficial to the subsequent application in the medicine for treating heart failure. The extraction method of the invention increases the extraction rate, improves the curative effect and has excellent safety.
The toad venom extract can obviously increase the cell survival rate, obviously improve the oxidation resistance of cells and further greatly improve the heart failure resistance of the medicament.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below with reference to specific embodiments, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The preparation method of the venenum bufonis extract comprises the following steps:
the method comprises the following steps: soaking venenum Bufonis powder with softening agent, and standing by after treatment;
step two: sending into 10-15 weight times of ethanol solvent for refluxing, removing ethanol, filtering to obtain first extractive solution, and collecting residue;
step three: performing supercritical extraction treatment on the filter residue to obtain a second extracting solution after extraction is finished;
step four: mixing the first extract and the second extract to obtain an extract, and finally feeding the extract into macroporous resin for elution;
step five: adding 5-10% of regulating agent into Bufonis venenum extract, stirring, and mixing to obtain Bufonis venenum extract.
The preparation method of the softening agent in this example is as follows: adding chitosan into 3-6 times of sodium alginate solution, stirring and dispersing fully, then adding modified zeolite powder accounting for 5-10% of the total amount of chitosan, stirring and mixing fully to obtain a softening agent;
the preparation method of the modified zeolite powder comprises the following steps:
feeding the zeolite powder of 150-200 meshes into a heat treatment at 300-400 ℃ for 20-30min, then cooling to room temperature at the speed of 1-3 ℃/min, then feeding into a hydrochloric acid solution of 3-6 times of mass fraction 5-10% to fully disperse, and washing and drying to obtain the modified zeolite powder.
The temperature of the infiltration treatment of the embodiment is 40-45 ℃, the treatment rotating speed is 350-450r/min, and the treatment time is 20-30min.
The mass fraction of the sodium alginate solution in the embodiment is 10-15%.
The reflux extraction time of the embodiment is 40-50min; the mass fraction of the refluxing ethanol solvent is 95-97%.
The supercritical extraction treatment of the embodiment has the extraction temperature of 38-42 ℃, the extraction pressure of 28-30MPa and the extraction time of 1-2h.
The specific operation steps of the macroporous resin elution treatment in this example are as follows:
and (3) loading the extracting solution on macroporous resin, removing impurities by elution, then eluting, and finally collecting the eluent, wherein the elution is carried out by adopting an ethanol solution with the mass fraction of 55-60%.
In the fifth step of this embodiment, the rotation speed for stirring and mixing uniformly is 550-650r/min, and the stirring time is 20-30min; the conditioning treatment agent comprises the following raw materials in parts by weight: 5-10 parts of lactose and 1-5 parts of microcrystalline cellulose.
The preparation method of the venenum bufonis extract of the embodiment is used for preparing the venenum bufonis extract.
The application of the venenum bufonis extract in preparing the medicine for treating heart failure is disclosed.
Example 1.
The preparation method of the venenum bufonis extract comprises the following steps:
the method comprises the following steps: soaking venenum Bufonis powder with softening agent, and standing by after treatment;
step two: sending into 10 weight times of ethanol solvent for refluxing, removing ethanol after refluxing extraction, filtering to obtain first extractive solution, and collecting residue;
step three: performing supercritical extraction treatment on the filter residue to obtain a second extracting solution after extraction is finished;
step four: mixing the first extract and the second extract to obtain an extract, and finally feeding the extract into macroporous resin for elution;
step five: adding 5% of conditioning agent into Bufonis venenum extract, stirring, and mixing to obtain Bufonis venenum extract.
The preparation method of the softening agent in this example is as follows: adding chitosan into 3 times of sodium alginate solution, stirring and dispersing fully, then adding modified zeolite powder accounting for 5% of the total amount of chitosan, stirring and mixing fully to obtain a softening agent;
the preparation method of the modified zeolite powder comprises the following steps:
and (2) feeding the 150-mesh zeolite powder into a furnace for heat treatment at 300 ℃ for 20min, then cooling to room temperature at the speed of 1 ℃/min, then feeding into 3 times of hydrochloric acid solution with the mass fraction of 5% for full dispersion, and washing and drying to obtain the modified zeolite powder.
The temperature of the infiltration treatment in the embodiment is 40 ℃, the treatment rotating speed is 350r/min, and the treatment time is 20min.
The mass fraction of the sodium alginate solution in this example was 10%.
The reflux extraction time of this example was 40min; the mass fraction of the refluxing ethanol solvent is 95%.
The supercritical extraction temperature of the supercritical fluid extraction treatment in this example was 38 ℃, the extraction pressure was 28MPa, and the extraction time was 1h.
The specific operation steps of the macroporous resin for elution treatment in this embodiment are as follows:
and (3) loading the extracting solution on macroporous resin, removing impurities by elution, then eluting, and finally collecting the eluent, wherein the elution is carried out by adopting an ethanol solution with the mass fraction of 55%.
In the fifth step of this embodiment, the rotation speed for stirring and mixing uniformly is 550r/min, and the stirring time is 20min; the conditioning treatment agent comprises the following raw materials in parts by weight: 5 parts of lactose and 1 part of microcrystalline cellulose.
The venenum bufonis extract obtained by the preparation method of the venenum bufonis extract of the embodiment is provided.
The application of the venenum bufonis extract in preparing the medicine for treating heart failure is disclosed.
Example 2.
The preparation method of the venenum bufonis extract comprises the following steps:
the method comprises the following steps: soaking venenum Bufonis powder with softening agent, and standing by after treatment;
step two: sending into 15 weight times of ethanol solvent for refluxing, removing ethanol after refluxing extraction, filtering to obtain first extractive solution, and collecting residue;
step three: performing supercritical extraction treatment on the filter residue to obtain a second extracting solution after extraction is finished;
step four: mixing the first extract and the second extract to obtain an extract, and finally feeding the extract into macroporous resin for elution;
step five: adding 10% of regulating agent into Bufonis venenum extract, stirring, and mixing to obtain Bufonis venenum extract.
The preparation method of the softening agent in this example is as follows: adding chitosan into 6 times of sodium alginate solution, stirring and dispersing fully, then adding modified zeolite powder accounting for 10% of the total amount of the chitosan, stirring and mixing fully to obtain a softening agent;
the preparation method of the modified zeolite powder comprises the following steps:
and (2) feeding the zeolite powder of 200 meshes into a reactor at 400 ℃ for heat treatment for 30min, then cooling to room temperature at the speed of 3 ℃/min, then feeding into a hydrochloric acid solution with the mass fraction of 10% which is 6 times that of the zeolite powder for full dispersion, and washing and drying to obtain the modified zeolite powder.
The temperature of the infiltration treatment in the embodiment is 45 ℃, the treatment rotating speed is 450r/min, and the treatment time is 30min.
The mass fraction of the sodium alginate solution in this example was 15%.
The reflux extraction time in this example was 50min; the mass fraction of the refluxing ethanol solvent is 97%.
The extraction temperature of the supercritical extraction treatment in this example was 42 ℃, the extraction pressure was 30MPa, and the extraction time was 2 hours.
The specific operation steps of the macroporous resin elution treatment in this example are as follows:
and (3) sampling the extracting solution on macroporous resin, removing impurities by adopting elution, then eluting, and finally collecting the eluent, wherein the elution is carried out by adopting an ethanol solution with the mass fraction of 60%.
In the fifth step of this embodiment, the rotation speed of stirring and mixing is 650r/min, and the stirring time is 30min; the conditioning treatment agent comprises the following raw materials in parts by weight: 10 parts of lactose and 5 parts of microcrystalline cellulose.
The preparation method of the venenum bufonis extract of the embodiment is used for preparing the venenum bufonis extract.
The application of the venenum bufonis extract in preparing the medicine for treating heart failure is disclosed.
Example 3.
The preparation method of the venenum bufonis extract comprises the following steps:
the method comprises the following steps: soaking venenum Bufonis powder with softening agent, and standing by after treatment;
step two: sending into 12 weight times of ethanol solvent for refluxing, removing ethanol after refluxing extraction, filtering to obtain first extractive solution, and collecting residue;
step three: performing supercritical extraction treatment on the filter residue to obtain a second extracting solution after extraction is finished;
step four: mixing the first extract and the second extract to obtain an extract, and finally feeding the extract into macroporous resin for elution;
step five: adding 7.5% of the total amount of the conditioning agent into the Bufonis venenum extract, and stirring and mixing to obtain Bufonis venenum extract.
The preparation method of the softening agent in this example is as follows: adding chitosan into 4.5 times of sodium alginate solution, stirring and dispersing fully, then adding modified zeolite powder accounting for 7.5% of the total amount of the chitosan, stirring and mixing fully to obtain a softening agent;
the preparation method of the modified zeolite powder comprises the following steps:
and (2) feeding the 175-mesh zeolite powder to 350 ℃ for heat treatment for 25min, then cooling to room temperature at the speed of 2 ℃/min, then feeding the zeolite powder to 4.5 times of hydrochloric acid solution with the mass fraction of 7.5% for full dispersion, and washing and drying the zeolite powder to obtain the modified zeolite powder.
The temperature of the infiltration treatment in this embodiment is 42 ℃, the treatment rotation speed is 390r/min, and the treatment time is 25min.
The mass fraction of the sodium alginate solution in this example was 12.5%.
The extraction time of the reflux in this example was 45min; the mass fraction of the refluxing ethanol solvent is 96%.
The supercritical extraction temperature of the supercritical fluid extraction treatment in this example was 40 ℃, the extraction pressure was 29MPa, and the extraction time was 1.5h.
The specific operation steps of the macroporous resin elution treatment in this example are as follows:
and (3) loading the extracting solution on macroporous resin, removing impurities by elution, then eluting, and finally collecting the eluent, wherein the elution is carried out by adopting an ethanol solution with the mass fraction of 59%.
In the fifth step of this embodiment, the rotation speed of stirring and mixing uniformly is 600r/min, and the stirring time is 25min; the conditioning treatment agent comprises the following raw materials in parts by weight: 7.5 parts of lactose and 3 parts of microcrystalline cellulose.
The venenum bufonis extract obtained by the preparation method of the venenum bufonis extract of the embodiment is provided.
The application of the venenum bufonis extract in preparing the medicine for treating heart failure is disclosed.
Example 4.
The preparation method of the venenum bufonis extract comprises the following steps:
the method comprises the following steps: soaking venenum Bufonis powder with softening agent, and standing by after treatment;
step two: sending into 12 weight times of ethanol solvent for refluxing, removing ethanol after refluxing extraction, filtering to obtain first extractive solution, and collecting residue;
step three: performing supercritical extraction treatment on the filter residue to obtain a second extracting solution after extraction is finished;
step four: mixing the first extract and the second extract to obtain an extract, and finally feeding the extract into macroporous resin for elution;
step five: adding 6% of conditioning agent into Bufonis venenum extract, stirring, and mixing to obtain Bufonis venenum extract.
The preparation method of the softening agent in this embodiment comprises: adding chitosan into 4 times of sodium alginate solution, stirring and dispersing fully, then adding modified zeolite powder accounting for 6% of the total amount of chitosan, stirring and mixing fully to obtain a softening agent;
the preparation method of the modified zeolite powder comprises the following steps:
and (2) feeding the 160-mesh zeolite powder to 320 ℃ for heat treatment for 22min, then cooling to room temperature at the speed of 2 ℃/min, then feeding the zeolite powder to 4 times of hydrochloric acid solution with the mass fraction of 6% for full dispersion, and washing and drying the zeolite powder to obtain the modified zeolite powder.
The temperature of the infiltration treatment in the embodiment is 42 ℃, the treatment rotating speed is 360r/min, and the treatment time is 22min.
The mass fraction of the sodium alginate solution in this example was 12%.
The extraction time of the reflux in this example was 42min; the mass fraction of the refluxing ethanol solvent is 96%.
The extraction temperature of the supercritical extraction treatment in this example was 39 ℃, the extraction pressure was 29MPa, and the extraction time was 1.2 hours.
The specific operation steps of the macroporous resin elution treatment in this example are as follows:
and (3) loading the extracting solution on macroporous resin, removing impurities by elution, then eluting, and finally collecting the eluent, wherein the elution is carried out by adopting an ethanol solution with the mass fraction of 58%.
In the fifth step of this embodiment, the rotation speed of stirring and mixing uniformly is 570r/min, and the stirring time is 22min; the conditioning treatment agent comprises the following raw materials in parts by weight: 6 parts of lactose and 2 parts of microcrystalline cellulose.
The preparation method of the venenum bufonis extract of the embodiment is used for preparing the venenum bufonis extract.
The application of the toad venom extract in preparing the medicine for treating heart failure is disclosed in the embodiment.
Comparative example 1.
Unlike example 3, no treatment with a softener was used.
Comparative example 2.
The difference from example 3 is that no modified zeolite powder was added in the preparation of the flexibilizer.
Comparative example 3.
The difference from example 3 is that the modified zeolite powder was replaced with zeolite.
Comparative example 4.
Unlike example 3, no supercritical extraction was used.
According to the invention, the extraction rate of bufogenin components is tested, and the performances of the extraction rates of the bufogenin components of examples 1-4 and comparative examples 1-4 are tested as follows:
as can be seen from comparative examples 1 to 4 and examples 1 to 4;
the preparation of the venenum bufonis extract is not soaked by the softening agent, the extraction rate of the product is remarkably reduced, the modified zeolite powder is not added in the preparation of the softening agent and is replaced by zeolite, the extraction rate of the product tends to be poor, meanwhile, the supercritical extraction treatment is not adopted, and the extraction rate of the product is further reduced.
The innovation points of the invention are as follows: the venenum bufonis extract is prepared by matching modified zeolite powder, wherein the solubility of chitosan is dispersed through sodium alginate, and the chitosan is modified and prepared through matching modified zeolite powder, the dispersity of the zeolite powder is enhanced after the zeolite powder is modified, and then the dispersing of the venenum bufonis powder is promoted in the softening agent, so that the synergistic extraction in subsequent reflux extraction and supercritical extraction is facilitated, and the extraction efficiency is improved; the conditioning agent is cooperated with lactose and microcrystalline cellulose, thereby being beneficial to the subsequent application in the medicine for treating heart failure.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
Claims (10)
1. The preparation method of the toad venom extract is characterized by comprising the following steps:
the method comprises the following steps: soaking venenum Bufonis powder with softening agent, and standing by after treatment;
step two: sending into 10-15 weight times of ethanol solvent for refluxing, removing ethanol, filtering to obtain first extractive solution, and collecting residue;
step three: performing supercritical extraction treatment on the filter residue to obtain a second extracting solution after extraction is finished;
step four: mixing the first extract and the second extract to obtain an extract, and finally feeding the extract into macroporous resin for elution;
step five: adding 5-10% of conditioning agent into Bufonis venenum extract, stirring, and mixing to obtain Bufonis venenum extract.
2. The method for preparing the venenum bufonis extract as claimed in claim 1, wherein the method for preparing the mollifying agent comprises the following steps: adding chitosan into 3-6 times of sodium alginate solution, stirring and dispersing fully, then adding modified zeolite powder accounting for 5-10% of the total amount of chitosan, stirring and mixing fully to obtain a softening agent;
the preparation method of the modified zeolite powder comprises the following steps:
feeding the zeolite powder of 150-200 meshes into a heat treatment at 300-400 ℃ for 20-30min, then cooling to room temperature at the speed of 1-3 ℃/min, then feeding into a hydrochloric acid solution of 3-6 times of mass fraction 5-10% to fully disperse, and washing and drying to obtain the modified zeolite powder.
3. The method for preparing a venenum Bufonis extract as claimed in claim 1, wherein the soaking treatment temperature is 40-45 deg.C, the treatment rotation speed is 350-450r/min, and the treatment time is 20-30min.
4. The preparation method of the toad venom extract as claimed in claim 2, wherein the mass fraction of the sodium alginate solution is 10-15%.
5. The method for preparing the venenum bufonis extract as claimed in claim 1, wherein the reflux extraction time is 40-50min; the mass fraction of the refluxing ethanol solvent is 95-97%.
6. The method for preparing the toad venom extract according to claim 1, wherein the supercritical extraction is carried out at an extraction temperature of 38-42 ℃ under an extraction pressure of 28-30MPa for an extraction time of 1-2h.
7. The preparation method of the toad venom extract according to claim 1, wherein the macroporous resin is eluted by the following steps:
and (3) loading the extracting solution on macroporous resin, removing impurities by elution, then eluting, and finally collecting the eluent, wherein the elution is carried out by adopting an ethanol solution with the mass fraction of 55-60%.
8. The method for preparing the toad venom extract according to claim 1, wherein the rotation speed for uniformly stirring and mixing in the fifth step is 550-650r/min, and the stirring time is 20-30min; the conditioning treatment agent comprises the following raw materials in parts by weight: 5-10 parts of lactose and 1-5 parts of microcrystalline cellulose.
9. A venenum bufonis extract obtained by the preparation method of a venenum bufonis extract as claimed in any one of claims 1-8.
10. Use of the bufonis venenum extract as claimed in claim 9 in the preparation of a medicament for the treatment of heart failure.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211220024.9A CN115282169B (en) | 2022-10-08 | 2022-10-08 | Bufonis venenum extract, and its preparation method and application in preparing medicine for treating heart failure |
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