CN115364149A - Preparation method and application of XinnaoUNIC capsule - Google Patents
Preparation method and application of XinnaoUNIC capsule Download PDFInfo
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- CN115364149A CN115364149A CN202211043751.2A CN202211043751A CN115364149A CN 115364149 A CN115364149 A CN 115364149A CN 202211043751 A CN202211043751 A CN 202211043751A CN 115364149 A CN115364149 A CN 115364149A
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- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
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- A61K36/18—Magnoliophyta (angiosperms)
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Abstract
The invention relates to the field of biological medicine, and particularly provides a preparation method of a Xinnaotong capsule, wherein the Xinnaotong capsule is prepared from erigeron breviscapus, giant knotweed rhizome, wild hawthorn fruit, persimmon leaf, acanthopanax senticosus, kudzu root and red sage root, and the preparation method comprises the steps of firstly extracting the giant knotweed rhizome, the red sage root and the acanthopanax senticosus by using an aqueous solution containing ethanol, separating to obtain an alcohol extract and medicinal dregs, and then extracting the medicinal dregs by using water, so that the content of resveratrol, tanshinone and salvianolic acid B in the Xinnaotong capsule is improved, and a hyperlipemia APOE mouse test proves that the Xinnaotong capsule prepared by the preparation method provided by the invention has the effect of reducing the content of TC and TG in serum; the invention also provides application of the Xinnaotong capsule prepared by the preparation method of the Xinnaotong capsule as a medicament for reducing the content of TC and TG in serum.
Description
Technical Field
The invention belongs to the field of biological medicines, and particularly relates to a preparation method and application of a Chinese patent medicine Xinnaotong capsule.
Background
The capsule is prepared from herba Erigerontis, rhizoma Polygoni Cuspidati, fructus crataegi, folium kaki, radix Acanthopanacis Senticosi, radix Puerariae and Saviae Miltiorrhizae radix, and has effects of promoting blood circulation for removing blood stasis, dredging collaterals and relieving pain. Can be used for treating thoracic obstruction, vertigo, chest distress, chest pain, palpitation, dizziness, headache, tinnitus, coronary heart disease, angina pectoris, cerebral arteriosclerosis and hyperlipidemia caused by blood stasis obstruction. The research on the heart-brain communicating capsules is less, the existing quality standard of the heart-brain communicating capsules, namely the standard number WS-10031 (ZD-0031) -2002 of the State drug administration of the people's republic of China, is provided by the Guizhou Taihe pharmaceutical Co., ltd, is rechecked by the drug inspection institute of Hubei province, is released by the State drug administration of the people's republic of China, is tried out and implemented 12 months and 1 day in 2002, and has been carried out for 20 years so far.
In the 20 years of pharmaceutical technology development and clinical application process of the capsule with heart and brain communication, the research summary of a certain medicinal material in the capsule with heart and brain communication is disclosed in the research progress of mechanism of water-soluble components of red sage root affecting bone metabolism disclosed by the Li Shuhui et al, the research progress of chemical components and pharmacological action of erigeron breviscapus disclosed by the Guoshen et al, and the research progress of chemical components, pharmacological action and modern clinical application research progress of acanthopanax senticosus leaf disclosed by the Huang Xiao Wei et al; shenyang pharmaceutical university Koelreuterian 2008 published quality control method and pharmacokinetics research of Xinnaoton capsules further research on the quality control method of Xinnaoton capsules is disclosed in the Master thesis, but deep research on the preparation method of the Xinnaoton capsules is less.
Disclosure of Invention
The invention provides a preparation method of a Xinnaotong capsule, the Xinnaotong capsule is prepared from erigeron breviscapus, giant knotweed rhizome, wild hawthorn fruit, persimmon leaf, acanthopanax, kudzu root and red sage root, and the preparation method comprises the following steps: step A, extracting giant knotweed rhizome, red sage root and acanthopanax root with an aqueous solution containing ethanol, and separating to obtain an ethanol extract and dregs; step B, extracting the dregs of a decoction with water, and separating to obtain a water extract and dregs of a decoction; step C, concentrating the alcohol extract and the water extract respectively, mixing, drying in vacuum, and crushing to obtain paste powder; step D, extracting erigeron breviscapus, wild hawthorn, persimmon leaves and kudzu roots with water, concentrating the extracting solution, drying in vacuum, and crushing to obtain paste powder; and E, combining the paste powder obtained in the step C and the paste powder obtained in the step D to obtain medicinal powder, adding auxiliary materials, uniformly mixing and then encapsulating.
On the basis of the existing preparation method, firstly, extracting polygonum cuspidatum, salvia miltiorrhiza and acanthopanax senticosus with an ethanol-containing aqueous solution, separating to obtain an ethanol extract and medicine residues, then, extracting the medicine residues with water, improving the content of resveratrol, tanshinone and salvianolic acid B in the capsule with the heart-brain communication through the two steps of fixing the sequence, and proved by a hyperlipemia APOE mouse test, the capsule has the effect of reducing the content of TC and TG in serum. On the basis of the existing preparation method, a person skilled in the art can aim at improving the content of resveratrol, tanshinone and salvianolic acid B in the capsule for cardio-cerebral concatevation, and on the basis of the two steps, other steps are selected from the prior art and combined with the steps to prepare the capsule for cardio-cerebral concatevation, so that the content of resveratrol, tanshinone and salvianolic acid B in the capsule for cardio-cerebral concatevation is improved. The heart-brain communication capsule prepared by the preparation method of the heart-brain communication capsule provided by the invention has the efficacy of reducing the content of TC and TG in serum as proved by a hyperlipemia APOE mouse test.
In a preferred embodiment of the present invention, the combination of the step of extracting polygonum cuspidatum, salvia miltiorrhiza and acanthopanax senticosus with an ethanol-containing aqueous solution and the steps B, C, D and E can significantly increase the content of resveratrol, tanshinone and salvianolic acid B in the capsule for cardio-cerebral-intercommunion. Particularly, the applicant is summarized through a large number of experiments, and when the step a is combined with the step B, namely that the contents of resveratrol, tanshinone and salvianolic acid B in the capsule for treating cardio-cerebral-pulmonary diseases can be obviously improved by firstly extracting the polygonum cuspidatum, the salvia miltiorrhiza and the acanthopanax senticosus with an ethanol-containing aqueous solution and then extracting the medicine residues with water. The heart-brain communication capsule prepared by the preparation method of the heart-brain communication capsule provided by the invention has the efficacy of reducing the content of TC and TG in serum as proved by a hyperlipemia APOE mouse test.
In a preferred embodiment of the present invention, the step C is to separately concentrate the alcohol extract and the water extract, mix them, dry them in vacuum, and pulverize them. In order to obtain the preferred implementation step C of the present invention, when considering whether the alcoholic extractive solution prepared in step a and the aqueous extractive solution prepared in step B are "mixed after concentration" or "mixed before concentration", the applicant has concluded through a large number of experiments that the content of resveratrol, tanshinone and salvianolic acid B in the capsule prepared by the preparation method including the operation step "mixed after concentration" is higher.
In a non-limiting embodiment of the present invention, in the step D, erigeron breviscapus, wild hawthorn, persimmon leaf and kudzu root are extracted with water, and the extract is concentrated, vacuum-dried and pulverized to obtain the paste powder. In this embodiment, step D follows the existing quality standards for extracting the four herbs, so as to reduce the cost of equipment modification and process control due to the change of the preparation method to the maximum extent.
In a non-limiting embodiment of the invention, the step E is to combine the paste powders obtained in the steps C and D to obtain a medicinal powder, add the auxiliary materials, mix the mixture uniformly, and fill the mixture into capsules. In this embodiment, the proportion ratio of the powder to the auxiliary materials for encapsulating, the preparation process, and the specific test reagents, materials, conditions and parameters are not described in step E, and it is well known to those skilled in the art that after the powder is obtained, the capsule can be prepared without creative work according to the ratio specified by the existing quality standards and the pharmaceutical auxiliary materials commonly used in the art. Such as: and D, combining the paste powder obtained in the step C and the paste powder obtained in the step D to obtain medicinal powder, adding 1-4 per mill of magnesium stearate for mixing, filling the mixed medicinal powder into empty capsules by using a filling machine according to the standard that each capsule is filled with 0.4g +/-7.5 percent to prepare the Xinnaotong capsules, finally filling the Xinnaotong capsules into aluminum-plastic plates, and packaging the Xinnaotong capsules into finished products through the procedures of boxing, code scanning, boxing and the like.
In a preferred embodiment of the present invention, the ethanol-containing aqueous solution is an aqueous solution having a concentration of 40 to 90% by volume. In the above embodiment that does not provide specific concentration of the aqueous solution of ethanol, those skilled in the art can use an aqueous solution with any concentration of ethanol to achieve the purpose of extracting polygonum cuspidatum, salvia miltiorrhiza and acanthopanax senticosus, and at the same time, those skilled in the art know that the higher the concentration of ethanol, the higher the cost, the more dangerous in the actual production process, therefore, the applicant has concluded through a large number of experiments that it is known that when the aqueous solution of ethanol is an aqueous solution containing 40-90% volume concentration, various components in polygonum cuspidatum, salvia miltiorrhiza and acanthopanax senticosus can be extracted at higher extraction rate while ensuring lower cost and controllable risk, that is, the content of resveratrol, tanshinone and salvianolic acid B in the capsule for cardio-cerebral conciliation can be increased.
In a preferred embodiment of the present invention, the amount of water extracted with water in step B is 4 times of the amount of the medicinal materials, and the extraction time is 1 hour.
In a preferred embodiment of the present invention, the number of times of water extraction in step D is two, wherein the water consumption in the first time is 6 times of the amount of the medicinal materials, and the water consumption in the second time is 4 times of the amount of the medicinal materials.
In a preferred embodiment of the invention, the concentration is such as to obtain an extract with a specific gravity of 1.25-1.30 (80 ℃).
In a preferred embodiment of the present invention, the pulverization is to 60 mesh.
In a preferred embodiment of the invention, the amount of the ethanol-containing aqueous solution is 6 times of the amount of the medicinal materials, the extraction temperature is less than or equal to 85 ℃, and the extraction time is 3-7 hours.
In a preferred embodiment of the invention, the content of resveratrol in the Xinnaotong capsule is not less than 1.62 mg/g. Because the preparation method of the capsule for treating cardio-cerebral-pulmonary diseases provided by the invention comprises the step of extracting giant knotweed rhizome, red sage root and acanthopanax root with an aqueous solution containing ethanol, the content of resveratrol in the prepared capsule for treating cardio-cerebral-pulmonary diseases is not lower than 1.62 mg/g.
In a preferred embodiment of the invention, the content of tanshinone in the capsule for treating cardio-cerebral concussion is not less than 1.67 mg/g. Because the preparation method of the capsule for treating cardio-cerebral-intercommunion provided by the invention comprises the step of extracting giant knotweed rhizome, red sage root and acanthopanax root by using an aqueous solution containing ethanol, the content of tanshinone in the prepared capsule for treating cardio-cerebral-intercommunion is not lower than 1.67 mg/g.
In a preferred embodiment of the invention, the content of salvianolic acid B in the capsule for treating cardio-cerebral communication is not less than 21.78 mg/g. Because the preparation method of the capsule for treating cardio-cerebral diseases provided by the invention comprises the step of extracting giant knotweed rhizome, salvia miltiorrhiza and acanthopanax root with an aqueous solution containing ethanol, the content of salvianolic acid B in the prepared capsule for treating cardio-cerebral diseases is not lower than 21.78 mg/g.
The invention also provides application of the capsule prepared by the preparation method of the capsule for heart-brain communication as a medicament for reducing the content of TC and TG in serum.
The preparation method of the heart-brain linking capsule provided by the invention comprises the two steps of firstly extracting giant knotweed rhizome, red sage root and acanthopanax root by using an aqueous solution containing ethanol, separating to obtain an ethanol extract and dregs, then extracting the dregs by using water, and fixing the sequence, so that the content of resveratrol, tanshinone and salvianolic acid B in the heart-brain linking capsule is improved, and the hyperlipemia APOE mouse test proves that the heart-brain linking capsule has the effect of reducing the content of TC and TG in serum. In a preferred embodiment of the present invention, the preparation method of the capsule for cardio-cerebral circulation provided by the present invention combines the step of extracting polygonum cuspidatum, salvia miltiorrhiza and acanthopanax senticosus with an aqueous solution containing ethanol, and separating to obtain an ethanol extract and dregs with the above step B, step C, step D and step E, so that the content of resveratrol, tanshinone and salvianolic acid B in the capsule for cardio-cerebral circulation can be significantly increased. The heart-brain connecting capsule prepared by the preparation method of the heart-brain connecting capsule provided by the invention has the effect of reducing the content of TC and TG in serum proved by hyperlipemia APOE mouse tests.
Drawings
Fig. 1 is a flow chart of a preparation method of the heart-brain communication capsule provided by the invention;
FIG. 2 is a chromatogram for measuring the content of resveratrol in the content of the capsule prepared in example 5;
fig. 3 is a chromatogram for measuring the tanshinone content in the capsule prepared in example 4 of the present invention;
fig. 4 is a chromatogram for measuring the content of salvianolic acid B in the content of the capsule prepared in example 3.
Detailed Description
In order to accurately represent and explain the technical solutions provided by the present invention, the technical terms used in the present invention are explained as follows before the summary of the invention.
The invention discloses a heart-brain communication capsule, which is characterized in that: is prepared from herba Erigerontis, rhizoma Polygoni Cuspidati, fructus crataegi Pinnatifidae, folium kaki, radix Acanthopanacis Senticosi, radix Puerariae and Saviae Miltiorrhizae radix.
The "preparation method of the heart-brain communication capsule" of the present invention refers to the preparation method claimed in the present invention, except that the preparation method is described in the quality standard.
The quality standard of the invention refers to: standard No. WS-10031 (ZD-0031) -2002, which is a standard listed in the national drug administration for people's republic of China.
The "aqueous solution of ethanol" in the invention refers to an aqueous solution containing ethanol; the concentrations of the solutions according to the invention are, unless otherwise specified, volume concentrations.
The dosage of the solvent used for extracting the medicinal materials by the solvent is the mass-volume ratio. For example: the method for extracting the medicinal material with the ethanol-containing aqueous solution with the amount of 6 times of the medicinal material comprises adding 6L of ethanol-containing aqueous solution into 1kg of the medicinal material; another example is: the step B, wherein the amount of water extracted by water is 4 times of the amount of the medicinal materials, is to add 4L of water to 1kg of the medicinal materials, and the other steps are the same.
Non-limiting embodiments of the present invention are further described below in conjunction with the appended drawings. It should be noted that the following embodiments are only illustrative and should not be construed as limiting the technical scope of the present invention.
Example 1
The embodiment illustrates a preparation method of a capsule for heart-brain communication, which is prepared from erigeron breviscapus, giant knotweed rhizome, wild hawthorn fruit, persimmon leaf, acanthopanax senticosus, kudzuvine root and red sage root, and the preparation method comprises the step of extracting the giant knotweed rhizome, the red sage root and the acanthopanax senticosus with an ethanol-containing aqueous solution according to the quality standard specified amount of the capsule for heart-brain communication, and then preparing the capsule powder for heart-brain communication. Finally, the medicinal powder is prepared into capsules according to the conventional preparation process.
Other test reagents, materials, conditions and parameters not described in this example can be selected by those skilled in the art with the creative efforts of referring to FIG. 1, and the specific steps can be implemented by selecting the reagents, materials, parameters and the implementation sequence. Wherein the capsule is prepared from herba Erigerontis, rhizoma Polygoni Cuspidati, fructus crataegi Pinnatifidae, folium kaki, radix Acanthopanacis Senticosi, radix Puerariae and Saviae Miltiorrhizae radix, and is prescribed by the standards of WS-10031 (ZD-0031) -2002 of the State drug administration of the people's republic of China. Based on the existing quality standard, the applicant is concluded through a large number of experiments that the content of resveratrol, tanshinone and salvianolic acid B in the capsule can be effectively improved when the preparation method of the capsule for treating cardio-cerebral vascular diseases comprises the step of extracting giant knotweed rhizome, salvia miltiorrhiza and acanthopanax senticosus with an ethanol-containing aqueous solution.
Example 2
In this embodiment, a preparation method of a capsule for cardio-cerebral vascular diseases is illustrated, based on embodiment 1, according to the specified quality standard of the capsule for cardio-cerebral vascular diseases, 390kg of giant knotweed rhizome, 320kg of acanthopanax root and 320kg of salvia miltiorrhiza are taken, 90% ethanol with 6 times of the amount of the medicinal materials is added for reflux extraction, the extraction temperature is controlled at 65 ℃, and the extraction time is 7 hours. Recovering ethanol from the extractive solution, concentrating to specific gravity of 1.27 (80 deg.C), and making into extract. After the ethanol is completely removed from the dregs after the ethanol extraction, adding water with the amount 4 times of the amount of the medicinal materials for decocting and extracting for 1 hour, separating the extracting solution, discarding the dregs, and concentrating to obtain an extract with the specific gravity of 1.30 (80 ℃) for later use. Mixing the ethanol extract and water extract, vacuum drying (at temperature below 75 deg.C) to obtain dry extract, and pulverizing into 60 mesh powder. Taking 390kg of erigeron breviscapus, 390kg of wild hawthorn, 390kg of persimmon leaf and 320kg of kudzu root according to the specified quality standard of the Xinnaotong capsule, adding water for decoction and extraction twice, adding 6 times of water for decoction and extraction for 1.5 hours for the first time, adding 4 times of water for decoction and extraction for 1 hour for the second time, combining the two extracting solutions, concentrating to obtain an extract with the specific gravity of 1.25 (80 ℃), and performing vacuum drying (less than 78 ℃) to obtain dry paste, wherein the dry paste powder is 60-mesh paste powder for later use. Mixing the two parts of the 60-mesh paste powder to obtain the capsule powder for heart-brain communication. Finally, 1-4 per mill of magnesium stearate is added into the medicinal powder for mixing, and the mixed medicinal powder is filled into empty capsules by a filling machine according to the standard of 0.4g +/-7.5 percent of each capsule, so as to prepare the Xinnaotong capsules.
In the present embodiment, other test reagents, materials, conditions, and parameters are not described, and those skilled in the art can select the reagents, materials, parameters, and the specific steps to implement in a creative manner, so as to obtain the capsule with enhanced contents of resveratrol, tanshinone, and salvianolic acid B.
Example 3
In this embodiment, a preparation method of a capsule for cardio-cerebral vascular diseases is illustrated, based on embodiment 1, according to the specified quality standard of the capsule for cardio-cerebral vascular diseases, 390kg of giant knotweed rhizome, 320kg of acanthopanax root and 320kg of salvia miltiorrhiza are taken, 80% ethanol with 6 times of the amount of the medicinal materials is added for reflux extraction, the extraction temperature is controlled at 75 ℃, and the extraction time is 3 hours. Recovering ethanol from the extractive solution, concentrating to specific gravity of 1.29 (80 deg.C), and making into extract. After the ethanol is completely removed from the dregs after the ethanol extraction, adding water with the amount 4 times of the amount of the medicinal materials for decocting and extracting for 1 hour, separating the extracting solution, discarding the dregs, and concentrating to obtain an extract with the specific gravity of 1.29 (80 ℃) for later use. Mixing the ethanol extract and water extract, vacuum drying (below 78 deg.C) to obtain dry extract, and making into 60 mesh extract powder. According to the specified weight of the Xinnaotong capsule based on the quality standard, 390kg of erigeron breviscapus, 390kg of wild hawthorn, 390kg of persimmon leaf and 320kg of kudzu root are taken, water is added for decoction and extraction for two times, water with 6 times of the amount of the medicinal materials is added for the first time, the water is added for decoction and extraction for 1.5 hours, water with 4 times of the amount of the medicinal materials is added for decoction and extraction for 1 hour for the second time, the two extracting solutions are combined and concentrated to extract with the specific gravity of 1.230 (80 ℃), and then vacuum drying is carried out (less than 78 ℃) to obtain dry paste, and the dry paste powder is 60-mesh paste powder for standby. Mixing the two parts of the 60-mesh paste powder to obtain the capsule powder for heart-brain communication. Finally, 1-4 per mill of magnesium stearate is added into the medicinal powder for mixing, and the mixed medicinal powder is filled into empty capsules by a filling machine according to the standard of 0.4g +/-7.5 percent of each capsule, so as to prepare the Xinnaotong capsules.
In the present embodiment, other test reagents, materials, conditions, and parameters are not described, and those skilled in the art can select the reagents, materials, parameters, and the specific steps to implement in a creative manner, so as to obtain the capsule with enhanced contents of resveratrol, tanshinone, and salvianolic acid B.
Example 4
This example illustrates a preparation method of a capsule for cardio-cerebral concussion, which is based on example 1, according to the specified quality standard of the capsule for cardio-cerebral concussion, 390kg of polygonum cuspidatum, 320kg of acanthopanax, and 320kg of salvia miltiorrhiza are taken, 70% ethanol 6 times the amount of the medicinal materials is added for reflux extraction, the extraction temperature is controlled at 82 ℃, and the extraction time is 6 hours. Recovering ethanol from the extractive solution, concentrating to specific gravity of 1.30 (80 deg.C), and making into extract. After the ethanol is removed from the dregs after the ethanol extraction, adding water with the amount 4 times of the medicinal materials for decocting and extracting for 1 hour, separating the extracting solution, discarding the dregs, and concentrating to obtain an extract with the specific gravity of 1.30 (80 ℃) for later use. Mixing the ethanol extract and water extract, vacuum drying (below 80 deg.C) to obtain dry extract, and pulverizing into 60 mesh powder. Taking 390kg of erigeron breviscapus, 390kg of wild hawthorn, 390kg of persimmon leaf and 320kg of kudzu root according to the specified quality standard of the Xinnaotong capsule, adding water for decoction and extraction twice, adding 6 times of water for decoction and extraction for 1.5 hours for the first time, adding 4 times of water for decoction and extraction for 1 hour for the second time, combining the two extracting solutions, concentrating to obtain an extract with the specific gravity of 1.30 (80 ℃), and performing vacuum drying (less than 80 ℃) to obtain dry paste, wherein the dry paste powder is 60-mesh paste powder for later use. Mixing the two parts of the 60-mesh paste powder to obtain the capsule powder for heart-brain communication. Finally, 1-4 per mill of magnesium stearate is added into the medicinal powder for mixing, and the mixed medicinal powder is filled into empty capsules by a filling machine according to the standard of 0.4g +/-7.5 percent of each capsule, so as to prepare the Xinnaotong capsules.
In the present embodiment, other test reagents, materials, conditions, and parameters are not described, and those skilled in the art can select the reagents, materials, parameters, and the specific steps to implement in a creative manner, so as to obtain the capsule with enhanced contents of resveratrol, tanshinone, and salvianolic acid B.
Example 5
In this embodiment, a preparation method of a capsule for cardio-cerebral vascular diseases is illustrated, based on embodiment 1, according to the specified quality standard of the capsule for cardio-cerebral vascular diseases, 390kg of giant knotweed rhizome, 320kg of acanthopanax root and 320kg of salvia miltiorrhiza are taken, 40% ethanol with 6 times of the amount of the medicinal materials is added for reflux extraction, the extraction temperature is controlled at 85 ℃, and the extraction time is 7 hours. Recovering ethanol from the extractive solution, concentrating to specific gravity of 1.29 (80 deg.C), and making into extract. After the ethanol is completely removed from the dregs after the ethanol extraction, adding water with the amount 4 times of the amount of the medicinal materials for decocting and extracting for 1 hour, separating the extracting solution, discarding the dregs, and concentrating to obtain an extract with the specific gravity of 1.30 (80 ℃) for later use. Mixing the ethanol extract and water extract, vacuum drying (below 75 deg.C) to obtain dry extract, and making into 60 mesh extract powder. Taking 390kg of erigeron breviscapus, 390kg of wild hawthorn, 390kg of persimmon leaf and 320kg of kudzuvine root according to the specified quality standard of the Xinnaotong capsule, adding water for decoction and extraction twice, adding 6 times of water for decoction and extraction for 1.5 hours for the first time, adding 4 times of water for decoction and extraction for 1 hour for the second time, combining the two extracting solutions, concentrating to obtain an extract with the specific gravity of 1.29 (80 ℃), and then carrying out vacuum drying (less than 75 ℃) to obtain dry paste, wherein the dry paste powder is 60-mesh paste powder for later use. Mixing the two parts of the 60-mesh paste powder to obtain the capsule powder for heart-brain communication. Finally, 1-4 per mill of magnesium stearate is added into the medicinal powder for mixing, and the mixed medicinal powder is filled into empty capsules by a filling machine according to the standard of 0.4g +/-7.5 percent of each capsule, so as to prepare the Xinnaotong capsules.
In the present embodiment, other test reagents, materials, conditions, and parameters are not described, and those skilled in the art can select the reagents, materials, parameters, and the specific steps to implement in a creative manner, so as to obtain the capsule with enhanced contents of resveratrol, tanshinone, and salvianolic acid B.
Example 6
In this example, the content of the capsule prepared in example 3 and having a linkage with the heart and the brain is measured by high performance liquid chromatography (general rule 0512) in accordance with pharmacopoeia of the people's republic of China. The method specifically comprises the following steps:
chromatographic conditions and system applicability test: octadecylsilane chemically bonded silica is used as a filling agent; taking acetonitrile-0.2% acetic acid solution (25) as a mobile phase, wherein the column temperature is 35 ℃; the detection wavelength was 306nm. The number of theoretical plates is not less than 3000 calculated according to the peak of resveratrol.
Preparation of control solutions: accurately weighing appropriate amount of resveratrol reference substance, placing in brown measuring flask, and adding methanol to obtain solution containing 20 μ g per 1 ml.
Preparation of a test solution: taking contents of the Xinnaotong capsule, grinding the contents into powder, taking about 1g (about 1g of small test mixed powder and about 3g of large production extract powder), precisely weighing, placing the powder into a conical flask with a plug, precisely adding 50ml of methanol, sealing the plug, weighing the weight, carrying out ultrasonic treatment (power 400W and frequency 40 kHz) for 45 minutes, cooling, weighing again, complementing the lost weight with methanol, shaking up, filtering, and taking a subsequent filtrate to obtain the Xinnaotong capsule.
The content determination method comprises the following steps: precisely sucking 10 μ l of each of the reference solution and the sample solution, injecting into liquid chromatograph, and measuring.
The content determination result is as follows: the content of resveratrol in the test product of the Xinnaotong capsule is 1.80mg/g. Taking the cardio-cerebral communication capsule prepared in the embodiment 5 of the invention as an example, please refer to fig. 2 for a chromatogram for measuring the content of resveratrol in the content of the cardio-cerebral communication capsule, and it needs to be explained that: FIG. 2 is only one of typical content measurement chromatograms for the test sample.
The content determination method of this embodiment requires attention to: the control and sample are prepared fresh.
Example 7
In this example, the content of the capsule prepared in example 4 was measured for tanshinone content by high performance liquid chromatography (general guideline 0512) in pharmacopoeia of the people's republic of China. The method comprises the following specific steps:
chromatographic conditions and system applicability test: octadecylsilane chemically bonded silica is used as a filling agent; taking acetonitrile as a mobile phase A and 0.02% phosphoric acid solution as a mobile phase B, and carrying out gradient elution according to the specification in the following table; the column temperature is 20 ℃; the detection wavelength was 270nm. The number of theoretical plates is not less than 60000 calculated according to tanshinone IIA peak.
Preparation of control solutions: taking a proper amount of tanshinone IIA reference substance, precisely weighing, placing in a brown measuring flask, and adding methanol to obtain a solution containing 20 μ g of tanshinone IIA per 1 ml.
Preparation of a test solution: taking the contents of the Xinnaotong capsule, grinding the powder (passing through a third sieve), taking about 2g of the powder, precisely weighing the powder, placing the powder into a conical flask with a plug, precisely adding 50ml of methanol, sealing the plug, weighing the weight, carrying out ultrasonic treatment (power 140W and frequency 42 kHz) for 30 minutes, cooling the mixture, weighing the weight again, complementing the weight loss by using the methanol, shaking the mixture evenly, filtering the mixture, and taking a subsequent filtrate to obtain the Xinnaotong capsule.
The content determination method comprises the following steps: precisely sucking 10 μ l of each of the reference solution and the sample solution, injecting into liquid chromatograph, and measuring. And calculating the relative retention time of cryptotanshinone and tanshinone I by taking the tanshinone IIA reference substance as a reference and the corresponding peak as an S peak, wherein the relative retention time is within +/-5% of a specified value. The relative retention times and correction factors are shown in the following table:
and taking the peak area of the tanshinone IIA as a reference, and multiplying the reference by a correction factor respectively to calculate the contents of the cryptotanshinone, the tanshinone I and the tanshinone IIA.
The content determination result is as follows: in the test sample of the Xinnaotong capsule, the content of tanshinone is 1.85mg/g based on the total amount of tanshinone IIA (C19H 18O 3), cryptotanshinone (C19H 20O 3) and tanshinone I (C18H 12O 3). Taking the cardio-cerebral communicating capsule prepared in the embodiment 4 of the invention as an example, please refer to fig. 3 for a chromatogram for measuring the content of tanshinone in the content of the cardio-cerebral communicating capsule, and it needs to be explained that: FIG. 3 is only one of typical content measurement chromatograms for the test sample.
Example 8
In this example, the content of the capsule prepared in example 5, XINNAOUNICOM, was measured for salvianolic acid B by high performance liquid chromatography (general rule 0512) in pharmacopoeia of the people's republic of China. The method comprises the following specific steps:
chromatographic conditions and system applicability test: octadecylsilane chemically bonded silica is used as a filling agent; acetonitrile-0.1% phosphoric acid solution (22; the column temperature is 20 ℃; the flow rate was 1.2 ml per minute and the detection wavelength was 286 nm. The number of theoretical plates should not be less than 6000 calculated according to the salvianolic acid B peak.
Preparation of control solutions: taking a proper amount of salvianolic acid B reference substance, precisely weighing, and adding a methanol-water (8).
Preparing a test solution: taking the contents of the Xinnaotong capsule, grinding the contents into powder (passing through a No. three sieve), taking about 2g of the powder, precisely weighing the powder, placing the powder into a conical flask with a plug, precisely adding 50ml of a methanol-water (8).
The content determination method comprises the following steps: precisely sucking 10 μ l of each of the reference solution and the sample solution, injecting into liquid chromatograph, and measuring.
The content determination result is as follows: the content of salvianolic acid B (C36H 30O 16) in the XinnaoUnion capsule is 24.2 mg/g. Taking the capsule for cardio-cerebral communication prepared in embodiment 3 of the present invention as an example, the chromatogram for measuring the content of salvianolic acid B in the content of the capsule for cardio-cerebral communication is shown in fig. 4, which is to be described as follows: FIG. 4 is only one of typical content measurement chromatograms for the test sample.
Example 9
In this example, the content of resveratrol, tanshinone and salvianolic acid B in the capsule prepared in example 4 of the present invention is determined by the high performance liquid chromatography described in examples 6 to 8, in which the capsule is compared with the capsule prepared by the preparation method described in the existing quality standard, and the specific steps are as follows:
the preparation method recorded by the existing quality standard comprises the following steps: decocting fleabane 390g, giant knotweed rhizome 390g, wild haw 390g, persimmon leaf 390g, acanthopanax root 320g, kudzu vine root 320g and red sage root 320g, filtering, concentrating the filtrate into clear paste with relative density of 1.25-1.30 (80 deg.C), vacuum drying to obtain dry extract for later use. Reflux-extracting the rest radix Acanthopanacis Senticosi and Saviae Miltiorrhizae radix with 4.5 times of 75% ethanol for 2.5 hr, filtering, recovering ethanol from filtrate, concentrating to obtain fluid extract with relative density of 1.22-1.27 (80 deg.C), and vacuum drying to obtain dry extract. Pulverizing the above two dry extracts, sieving, mixing, and making into capsule.
Chromatographic conditions and system applicability test:
the content of resveratrol was measured by the content measurement method described in example 6;
the content determination method described in example 7 was used to determine the tanshinone content;
the content of salvianolic acid B was measured by the content measurement method described in example 8.
Preparation of control solutions:
reference solutions of resveratrol, tanshinone and salvianolic acid B were prepared according to the methods for preparing reference solutions described in examples 6, 7 and 8, respectively.
Preparation of a test solution:
taking the contents of the Xinnaotong capsule, grinding the contents into powder (passing through a No. three sieve), taking about 2g of the powder, precisely weighing the powder, placing the powder into a conical flask with a plug, precisely adding 50ml of a methanol-water (8).
Test solution 1: the capsules for cardio-cerebral concussion prepared in example 4 were prepared according to the method for preparing the test solution.
Sample solution 2: the preparation method of the test solution is used for preparing the Xinnaotong capsule prepared by the preparation method recorded in the existing quality standard.
The content determination method comprises the following steps: preparing 3 parts of each sample solution by the above method, respectively sucking 10 μ l of each of the reference solution, sample solution 1 and sample solution 2, injecting into liquid chromatograph, and measuring.
The content determination result is as follows:
from the content measurement results, it can be seen that the content of the resveratrol, the tanshinone and the salvianolic acid B in the capsule for cardio-cerebral concatevation prepared in the embodiment 4 of the invention is higher than that of the capsule for cardio-cerebral concatevation prepared by the preparation method recorded in the quality standard. Compared with the capsule prepared by the preparation method recorded in the quality standard, the capsule prepared in the embodiment 4 of the invention has the advantages that the content of resveratrol is improved by 309.1%, the content of tanshinone is improved by 179.5%, and the content of salvianolic acid B is improved by 6.2%.
Example 10
In this embodiment, when comparing the capsule for cardio-cerebral concussion prepared in example 4 of the present invention with the capsule for cardio-cerebral concussion prepared by other preparation methods explored during the process of obtaining the preparation method of the capsule for cardio-cerebral concussion of the present invention, the contents of resveratrol, tanshinone and salvianolic acid B are measured by the high performance liquid chromatography described in examples 6 to 8, specifically as follows:
other preparation methods of the heart-brain communication capsule comprise the following steps: according to the specified amount of the quality standard of the Xinnaotong capsule, 390kg of giant knotweed, 320kg of acanthopanax and 320kg of red sage root are taken, 70 percent ethanol with 6 times of the amount of the medicinal materials is added for reflux extraction, the extraction temperature is controlled at 80 ℃, and the extraction time is 3 hours. Recovering ethanol from the extractive solution, concentrating to specific gravity of 1.30 (80 deg.C), and making into extract. After the ethanol is removed from the dregs after the ethanol extraction, 70 percent ethanol with the amount 4 times of the amount of the medicinal materials is added for extraction for 1 hour, the extracting solution is separated, the dregs are discarded, and the dregs are concentrated to extract with the specific gravity of 1.30 (80 ℃) for standby. Mixing the two ethanol extracts, vacuum drying (below 75 deg.C) to obtain dry extract, and pulverizing into 60 mesh extract powder. Taking 390kg of erigeron breviscapus, 390kg of wild hawthorn, 390kg of persimmon leaf and 320kg of kudzu root according to the specified quality standard of the Xinnaotong capsule, adding water for decoction and extraction twice, adding 6 times of water for decoction and extraction for 2 hours for the first time, adding 4 times of water for decoction and extraction for 1 hour for the second time, combining the two extracting solutions, concentrating to obtain an extract with the specific gravity of 1.29 (80 ℃), and then carrying out vacuum drying (less than 75 ℃) to obtain a dry paste, and pulverizing into 60-mesh paste powder for later use. Mixing the two parts of the 60-mesh paste powder to obtain the capsule powder for heart-brain communication. Finally, 1-4 per mill of magnesium stearate is added into the medicinal powder for mixing, and the mixed medicinal powder is filled into empty capsules by a filling machine according to the standard of 0.4g +/-7.5 percent of each capsule, so as to prepare the Xinnaotong capsules.
Other preparation methods of the heart-brain communication capsule 2: taking 390kg of giant knotweed, 320kg of acanthopanax and 320kg of salvia miltiorrhiza according to the specified quality standard of the Xinnaotong capsule, adding 75% ethanol with 6 times of the amount of the medicinal materials for reflux extraction, controlling the extraction temperature at 80 ℃, extracting for 3 hours, and separating out the extracting solution. After ethanol is completely removed from the dregs after the ethanol extraction, adding water with the amount 4 times of the amount of the medicinal materials for decoction and extraction for 1 hour, separating the extracting solution, discarding the dregs, combining the extracting solutions (the ethanol extracting solution and the water extracting solution) twice, concentrating to obtain an extract with the specific gravity of 1.30 (80 ℃), and drying in vacuum (less than 75 ℃) to obtain dry paste, and pulverizing the dry paste into 60-mesh paste powder for later use. Taking 390kg of erigeron breviscapus, 390kg of wild hawthorn, 390kg of persimmon leaf and 320kg of kudzu root according to the specified quality standard of the Xinnaotong capsule, adding water for decoction and extraction twice, adding 6 times of water for decoction and extraction for 2 hours for the first time, adding 4 times of water for decoction and extraction for 1 hour for the second time, combining the two extracting solutions, concentrating to obtain an extract with the specific gravity of 1.30 (80 ℃), and then carrying out vacuum drying (less than 75 ℃) to obtain a dry paste, and pulverizing into 60-mesh paste powder for later use. Mixing the two parts of the 60-mesh paste powder to obtain the capsule powder for heart-brain communication. Finally, 1-4 per mill of magnesium stearate is added into the medicinal powder for mixing, and the mixed medicinal powder is filled into empty capsules by a filling machine according to the standard of 0.4g +/-7.5 percent of each capsule, so as to prepare the Xinnaotong capsules.
Chromatographic conditions and system applicability test:
the content of resveratrol was measured by the content measurement method described in example 6;
the content determination method described in example 7 was used to determine the tanshinone content;
the content of salvianolic acid B was measured by the method described in example 8.
Preparation of control solutions:
the reference solutions of resveratrol, tanshinone and salvianolic acid B were prepared according to the methods of preparation of the reference solutions described in examples 6, 7 and 8, respectively.
Preparation of a test solution:
taking the contents of the Xinnaotong capsule, grinding the capsule to powder (passing through a No. three sieve), taking about 2g, precisely weighing, placing the capsule in a conical flask with a plug, precisely adding 50ml of a methanol-water (8).
Test solution 1: the capsules for cardio-cerebral concussion prepared in example 4 were prepared according to the method for preparing the test solution.
Sample solution 2: the Xinnaotong capsule prepared by the other preparation method 1 of the Xinnaotong capsule is prepared according to the preparation method of the test solution.
Test solution 3: preparing the Xinnaotong capsule prepared by the other preparation methods 2 of the Xinnaotong capsule according to the preparation method of the test solution.
The content determination method comprises the following steps: preparing 3 parts of each sample solution by the above method, respectively sucking 10 μ l of each of the reference solution, sample solution 1 and sample solution 2, injecting into liquid chromatograph, and measuring.
The content determination result is as follows:
from the content measurement results, it can be seen that, compared with the capsule prepared by the other preparation method 1, the content of resveratrol in the capsule prepared in embodiment 4 of the present invention is increased by 3.4%, the content of salvianolic acid B is increased by 8.8%, and the content of tanshinone iia is decreased by 0.5%; compared with other cardio-cerebral communicated capsules prepared by the preparation method 2, the content of resveratrol in the cardio-cerebral communicated capsule prepared in the embodiment 4 of the invention is reduced by 1.6%, the content of salvianolic acid B is improved by 160.3%, and the content of tanshinone IIA is reduced by 0.5%. The preparation method of the test sample 1 (i.e., the preparation method in embodiment 4 of the present invention) is determined to be a better method by comprehensively considering reagent cost and operation cost generated by performing two times of alcohol extraction on giant knotweed rhizome, acanthopanax root and red sage root in the other preparation methods 1 and comprehensively considering the increase range of the content of resveratrol, salvianolic acid B and tanshinone IIA.
Example 11
In this example, the two different preparation methods of the capsule for treating cardiovascular and cerebrovascular diseases prepared by the method are respectively used to study the pharmacodynamic changes of the hyperlipemia APOE mice.
Wherein, the capsule for heart and brain communication prepared by two different preparation methods is respectively:
the first method comprises the following steps: the capsule for treating cardio-cerebral diseases is prepared by the preparation method of the embodiment 3;
and the second method comprises the following steps: the capsule is prepared by the preparation method recorded by the quality standard of the XinnaoUNIC.
The specific implementation method comprises the following steps:
1. materials and methods
1.1 Test animals: SPF cleaning grade male healthy APOE mice 32, 6-7 weeks old, 18-22g in body mass. The test animals were provided by the laboratory animal technology, inc. of Weitonglihua, beijing.
1.2 Drugs and reagents: the capsule for cardio-cerebral communication prepared by the preparation method of invention example 3 (hereinafter referred to as "example 3 method group"); the capsule (hereinafter referred to as "quality standard method group") is prepared by the existing preparation method recorded by the quality standard of cardio-cerebral communication; TG, TC detect reagent box (purchased from Nanjing institute of bioengineering, inc.).
1.3 The instrument comprises the following steps: a centrifuge and a microplate reader.
1.4 Method of producing a composite material
1.4.1 modeling: the 32 APOE mice were randomly divided into normal group (8) and high fat group (24), and were all raised in a well ventilated room at 20-22 deg.C and 45-55% humidity, and the animals were allowed to drink water freely. Normal group mice were given normal diet, and high-fat group mice were given high-fat diet daily for 8 weeks. After 8 weeks of modeling, blood of each group of mice is collected by infraorbital veins of the mice, blood lipid indexes (TC and TG) are detected, and at least one blood lipid index is obviously different from that of a blank control group, namely the modeling is successful, and 24 hyperlipidemia mouse models are successfully prepared in the research.
1.4.2 administration: the 24 high-fat mice were randomly divided into 8 mice each of a model group, a quality standard method group (administered dose of 0.78 g/kg), and a method group of example 3 (administered dose of 0.78 g/kg). The normal group of mice was given no additional intervention, and the remaining groups of mice were given daily high-fat diet feeding and the corresponding drugs for 4 weeks.
1.4.3 specimen Collection: after all mice are dosed, the eyeballs are picked and blood is taken, after standing for 30min at room temperature, centrifugation is carried out for 30min at 2500r/min, and serum is collected and placed in a refrigerator at minus 80 ℃ for storage to be tested.
1.5 Statistical analysis: statistical analysis using GraphPad Prism7 software, data was measured to
Formally express, measure data and
the form is expressed, the single-factor variance analysis is adopted for the comparison measurement data among a plurality of groups, and the T test is adopted for the pairwise comparison. P <0.05 indicates that the difference of the comparison results is statistically significant.
2. Test results
2.1 Serum TC and TG changes of each group of mice 8 weeks after model building
The blood lipid data result shows that the TC and TG levels in the serum of the high-fat feed group are obviously increased (P is less than 0.01) compared with the blank control group after 8 weeks of molding.
2.2 Serum TC, TG changes in groups of mice 4 weeks after administration
The blood lipid data result shows that after 4 weeks of administration, compared with the model group, the TC level in the serum of the quality standard method group is remarkably reduced (P < 0.05), the TG level has no remarkable difference but has a callback effect, and the TC level and the TG level in the serum of the method group of the example 3 are both remarkably reduced (P < 0.01).
3. Conclusion of the experiment
The test results show that the Xinnaotong capsule has better blood fat reducing effect, the sample of the Xinnaotong capsule prepared by the preparation method of the embodiment 3 can obviously improve the content of TC and TG in serum, and the lipid reducing effect is better than that of the Xinnaotong capsule prepared by the existing preparation method recorded by the quality standard of the Xinnaotong.
The contents of resveratrol, salvianolic acid and tanshinone ia in the capsules for cardio-cerebral concateurs prepared by the preparation method described in the existing quality standard of the capsules for cardio-cerebral concateurs were compared with the contents of resveratrol, salvianolic acid and tanshinone ia in the capsules for cardio-cerebral concateurs prepared in example 9 and example 10. It should be noted that the applicant has obtained almost the same experimental results after performing the above experiments using the pluripotent vascular progenitor cells provided by the present invention including the methods of examples 1 to 5.
In the above example 11, the pharmacodynamic study of the hyperlipemia APOE mouse was performed by using the cardiocerebral communicating capsule prepared in example 3 as a representative. It should be noted that the applicant has obtained almost the same experimental results after performing the above experiments using the pluripotent vascular progenitor cells provided by the present invention including the methods of examples 1 to 5.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the technical scope of the present invention, and any person skilled in the art should be able to substitute or change the technical idea of the present invention within the technical scope of the present invention.
Claims (9)
1. The preparation method of the Xinnaotong capsule is characterized by comprising the following steps of:
step A, extracting giant knotweed rhizome, red sage root and acanthopanax root with an aqueous solution containing ethanol, and separating to obtain an ethanol extract and dregs of a decoction;
step B, extracting the dregs of a decoction with water, and separating to obtain a water extract and dregs of a decoction;
step C, concentrating the alcohol extract and the water extract respectively, mixing, drying in vacuum, and crushing to obtain paste powder;
step D, extracting erigeron breviscapus, wild hawthorn, persimmon leaves and kudzu roots with water, concentrating the extracting solution, drying in vacuum, and crushing to obtain paste powder;
and E, combining the paste powder obtained in the step C and the paste powder obtained in the step D to obtain medicinal powder, adding auxiliary materials, uniformly mixing and then encapsulating.
2. The method according to claim 1, wherein the aqueous solution containing ethanol is an aqueous solution having a concentration of 40-90% by volume.
3. The method according to claim 1, wherein the amount of the ethanol-containing aqueous solution is 6 times of the amount of the medicinal material, the extraction temperature is 85 ℃ or less, and the extraction time is 3-7 hours.
4. The method of claim 1, wherein the amount of water extracted with water in step B is 4 times the amount of the medicinal material, and the extraction time is 1 hour.
5. The method of claim 1, wherein the number of times of water extraction in step D is two, wherein the amount of water used in the first time is 6 times of the amount of the herb, and the amount of water used in the second time is 4 times of the amount of the herb.
6. The method according to claim 1, wherein the concentration is such as to obtain an extract having a specific gravity of 1.25-1.30 at 80 ℃.
7. The method of claim 1, wherein the pulverizing is to 60 mesh.
8. The method according to any one of claims 1 to 7, wherein the content of resveratrol in the capsule for cardio-cerebral communication is not less than 1.62 mg/g; or, the content of tanshinone in the capsule is not lower than 1.67 mg/g; or the content of the salvianolic acid B in the capsule for treating cardio-cerebral communication is not less than 21.78 mg/g.
9. Use of the Xinnaotong capsule prepared by the method of any one of claims 1 to 8 as a medicament for reducing the content of TC and TG in serum.
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| CN1490028A (en) * | 2003-04-01 | 2004-04-21 | 贵州太和制药有限公司 | Chinese medicinal preparations for cardiac and encephalic blood vessel diseases and productions thereof |
| CN101966239A (en) * | 2010-09-30 | 2011-02-09 | 贵州太和制药有限公司 | Chinese medicinal composition for preventing and treating cardiac cerebral and vascular diseases and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1490028A (en) * | 2003-04-01 | 2004-04-21 | 贵州太和制药有限公司 | Chinese medicinal preparations for cardiac and encephalic blood vessel diseases and productions thereof |
| CN101966239A (en) * | 2010-09-30 | 2011-02-09 | 贵州太和制药有限公司 | Chinese medicinal composition for preventing and treating cardiac cerebral and vascular diseases and preparation method thereof |
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| Title |
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| 苏子仁等: "虎杖提取工艺初探" * |
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