CN107823420A - The preparation method and Shenmai injection of Shenmai injection - Google Patents
The preparation method and Shenmai injection of Shenmai injection Download PDFInfo
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- CN107823420A CN107823420A CN201711101468.XA CN201711101468A CN107823420A CN 107823420 A CN107823420 A CN 107823420A CN 201711101468 A CN201711101468 A CN 201711101468A CN 107823420 A CN107823420 A CN 107823420A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
- A61K36/258—Panax (ginseng)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
- A61K36/8968—Ophiopogon (Lilyturf)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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Abstract
The present invention relates to pharmaceutical field, in particular to the preparation method and Shenmai injection of a kind of Shenmai injection.This method, in red ginseng extraction process, extract solution is concentrated into organic solvent-free, after moisturizing, regulating liquid medicine pH value, macromolecular precipitation is associated with so as to promote decoction small molecule, is associated with using macromolecular substances in milipore filter removing decoction and easily the small molecular weight impurity of macromolecular substances.Not only quality meets the Shenmai injection being prepared by this method《Chinese Pharmacopoeia》Four injection item items Testing index, and without polyoxyethylene sorbitan monoleate auxiliary material is added, so as to greatly reduce adverse reaction clinical safety hidden danger.
Description
Technical field
The present invention relates to pharmaceutical field, in particular to the preparation method and injection containing ginseng extract of a kind of Shenmai injection
Liquid.
Background technology
At present, the Shenmai injection finished product produced according to statutory standards preparation method, it is necessary to polyoxyethylene sorbitan monoleate auxiliary material is added,
Carry out in increase-volume decoction to be also easy to produce the material of visible foreign matters, if not adding polyoxyethylene sorbitan monoleate auxiliary material, final product quality is not met《China
Pharmacopeia》Four injection item visible foreign matters check criteria.
Polyoxyethylene sorbitan monoleate adds as solubilized auxiliary materials, but it is to cause traditional Chinese medicine adverse reaction to have the document report auxiliary material
One of the main reason for, and addition is related to adverse reaction.Therefore it is bad to there is introducing in the Shenmai injection of prior art
The risk of reaction source.
The content of the invention
, can be not add polyoxyethylene sorbitan monoleate auxiliary it is an object of the invention to provide a kind of preparation method of Shenmai injection
Under material, meet product quality《Chinese Pharmacopoeia》Four injection item items Testing index, can greatly reduce adverse reaction and face
Bed potential safety hazard.
The second object of the present invention is to provide a kind of Shenmai injection, can in the case where not adding polyoxyethylene sorbitan monoleate auxiliary material,
Quality meets《Chinese Pharmacopoeia》Four injection item items Testing index, can greatly reduce adverse reaction clinical safety hidden danger.
In order to realize the above-mentioned purpose of the present invention, spy uses following technical scheme：
A kind of preparation method of Shenmai injection, Shenmai injection are made up of red ginseng, the tuber of dwarf lilyturf and water；The system of Shenmai injection
Preparation Method comprises the following steps：Red ginseng is extracted with the first extractant, the solvent in the first extract solution is removed, adds water, obtain red ginseng and carry
Liquid is taken, adjusts the pH to 5.5~8.0 of red ginseng extract；The tuber of dwarf lilyturf is extracted with the second extractant, removed molten in the second extract solution
Agent, add water, obtain Radix Ophiopogonis extract, adjust the pH to 5.0~8.0 of Radix Ophiopogonis extract；Red ginseng extract and Radix Ophiopogonis extract are adopted
With merging after ultrafiltration membrance filter；First extractant and the second extractant are the mixture of organic solvent or organic solvent and water；
Any one of organic solvent in methanol, ethanol and ethyl acetate.
A kind of Shenmai injection, it is prepared by the preparation method of above-mentioned Shenmai injection.
Compared with prior art, beneficial effects of the present invention are：
The preparation method of Shenmai injection provided by the invention, comprises the following steps：Red ginseng is extracted with the first extractant, gone
Except the solvent in the first extract solution, add water, obtain red ginseng extract, adjust the pH to 5.5~8.0 of red ginseng extract；Carried with second
Take agent to extract the tuber of dwarf lilyturf, remove the solvent in the second extract solution, add water, obtain Radix Ophiopogonis extract, adjust the pH to 5.0 of Radix Ophiopogonis extract
~8.0；Merge red ginseng extract and Radix Ophiopogonis extract, add water, regulation pH value to 7.5~8.5, Shenmai injection is made.Using
Above-mentioned preparation method, in red ginseng extraction process, extract solution is concentrated into organic solvent-free, after moisturizing, regulating liquid medicine pH value, so as to
Promote decoction small molecule and be associated with macromolecular precipitation, remove macromolecular substances in decoction using milipore filter is associated with big point with easy
The small molecular weight impurity of sub- material, so that in the case where not adding polyoxyethylene sorbitan monoleate auxiliary material, the Shenmai injection that is prepared, quality
Meet《Chinese Pharmacopoeia》Four injection item items Testing index, and can greatly reduce adverse reaction clinical safety hidden danger.
Shenmai injection provided by the invention, it is prepared by the preparation method of above-mentioned Shenmai injection.The ginseng wheat is noted
Penetrating liquid, not only quality meets《Chinese Pharmacopoeia》Four injection item items Testing index, and it is auxiliary without polyoxyethylene sorbitan monoleate is added
Material, so as to greatly reduce adverse reaction clinical safety hidden danger.
Brief description of the drawings
For the technical scheme of the clearer explanation embodiment of the present invention, the accompanying drawing needed to use in embodiment is made below
It is simple to introduce.It should be appreciated that the following drawings illustrate only some embodiments of the present invention, it is not construed as to model of the present invention
The limitation enclosed.To those skilled in the art, without creative efforts, can be obtained according to these accompanying drawings
Obtain other accompanying drawings.
Fig. 1 is the finger-print of Shenmai injection provided in an embodiment of the present invention.
Embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the present invention.It is unreceipted specific in embodiment
Condition person, the condition suggested according to normal condition or manufacturer are carried out.Agents useful for same or the unreceipted production firm person of instrument, it is
The conventional products that can be obtained by commercially available purchase.
The present embodiment provides a kind of preparation method of Shenmai injection, and Shenmai injection is made up of red ginseng, the tuber of dwarf lilyturf and water；Ginseng
The preparation method of wheat parenteral solution comprises the following steps：
S1, with the first extractant extract red ginseng, remove the first extract solution in solvent, add water, obtain red ginseng extract, adjust
The pH of the red ginseng extract to 5.5~8.0.
In red ginseng extraction process, extract solution is concentrated into organic solvent-free, after moisturizing, by regulating liquid medicine pH value, can promote
Enter decoction small molecule and be associated with macromolecular precipitation, contribute in follow-up process that macromolecular precipitation filter off removing, so as to really
Protect the quality of medicine.
It should be understood that during solvent in the above-mentioned extract solution of removal first, solvent includes the first extractant and the first extractant
Solution, obtained red ginseng extract only include the decoction and water of remaining red ginseng medicinal material.
It is further preferred that the pH to 7.0 of regulation red ginseng extract, it is easier to promote decoction small molecule to be associated with macromolecular
Precipitation.
Further, extraction red ginseng is with the first extractant heating and refluxing extraction red ginseng 3~6 times.
Wherein, the first extractant is organic solvent.Organic solvent is any one in methanol, ethanol and ethyl acetate
Kind.
It is further preferred that above-mentioned organic solvent selection ethanol., can not only be effective when organic solvent selects ethanol
Ground extracts to red ginseng, and extraction effect is good, moreover, cost is relatively low, extraction is more prone to, extraction process safety.
Further preferably, the selection of times of refluxing extraction red ginseng 5 times.When the selection of times 5 times of refluxing extraction red ginseng
When, effectively red ginseng can not only be extracted, and extraction effect is good, will not cause the waste of resource.
It should be noted that during the above-mentioned progress refluxing extraction to red ginseng, heating-up temperature can be according to selected extractant
Type is selected.Further, the weight of each 1~2 hour extraction time, the first extractant and red ginseng than control 2~
5：1.So as to not extracted only effectively, and improve the recovery rate of the active component in raw material.
Further, when removing the solvent in the first extract solution activated carbon is additionally added to be adsorbed.
Adsorbed by adding activated carbon, can further remove the impurity in decoction, so as to obtain height to be follow-up
The medicine of quality provides favourable guarantee.
Specifically, after complete to red ginseng refluxing extraction, phegma is filtered, filtrate decompression is recycled to regulation crude drug amount, refrigerates,
Filtering.Then a certain amount of activated carbon is added, stirring certain time makes it fully adsorb, and filters.
It should be noted that being adsorbed to adding activated carbon in above-mentioned decoction, can be selected according to the situation of reality
Absorption is once or absorption is multiple.In the process, suitably moisturizing can be carried out to decoction according to being actually needed.Further
Ground, the selection of activated carbon can select the common activated carbon species applicatory in this area according to actual conditions.
Further, the pH for adjusting red ginseng extract is adjusted with inorganic alkali solution.
In optional embodiment of the present invention, inorganic alkali solution can select NaOH solution.
In other optional embodiments of the invention, above-mentioned inorganic alkali solution is also an option that other this areas are common
Aqueous slkali.For example, KOH solution.
It should be noted that the pH of red ginseng extract is adjusted with inorganic alkali solution to select regulation according to the situation of reality
Once or regulation is multiple, until the pH of decoction is adjusted to 5.0~8.0.In the process, can be according to being actually needed to medicine
Liquid carries out suitably moisturizing.
In optional embodiment of the present invention, adsorbed by adding activated carbon, regulating liquid medicine pH after filtration, repeat with
Upper process is twice.
S2, with the second extractant extract the tuber of dwarf lilyturf, remove the second extract solution in solvent, add water, obtain Radix Ophiopogonis extract, adjust
The pH of Radix Ophiopogonis extract to 5.0~8.0.
It should be understood that during solvent in the above-mentioned extract solution of removal second, the second extractant and the second extraction agent solution, obtain
Radix Ophiopogonis extract only include the decoction and water of remaining tuber of dwarf lilyturf medicinal material.
It is further preferred that the pH to 8.0 of regulation Radix Ophiopogonis extract.Decoction small molecule is further promoted to be associated with big point
Son precipitation, contribute in follow-up process that macromolecular precipitation filter off removing, so that it is guaranteed that the quality of medicine.
Further, the extraction tuber of dwarf lilyturf is with second extractant heating and refluxing extraction tuber of dwarf lilyturf 2~3 times.
Wherein, the second extractant is organic solvent.Organic solvent is any one in methanol, ethanol and ethyl acetate
Kind.
It is further preferred that above-mentioned organic solvent selection ethanol., can not only be effective when organic solvent selects ethanol
Ground extracts to the tuber of dwarf lilyturf, and extraction effect is good, moreover, cost is relatively low, extraction is more prone to, extraction process safety.
Further preferably, the selection of times of the refluxing extraction tuber of dwarf lilyturf 2 times.When the selection of times 2 times of the refluxing extraction tuber of dwarf lilyturf
When, effectively the tuber of dwarf lilyturf can not only be extracted, and extraction effect is good, will not cause the waste of resource.
It should be noted that during the above-mentioned progress refluxing extraction to the tuber of dwarf lilyturf, heating-up temperature can be according to selected extractant
Type is selected.Further, each 1~2 hour extraction time, the second extractant and the weight of the tuber of dwarf lilyturf than control 2~
5：1.So as to not extracted only effectively, and improve the recovery rate of the active component in raw material.
Further, when removing the solvent in the second extract solution activated carbon is additionally added to be adsorbed.
Adsorbed by adding activated carbon, can further remove the impurity in decoction, so as to obtain height to be follow-up
The medicine of quality provides favourable guarantee.
Specifically, after complete to tuber of dwarf lilyturf refluxing extraction, phegma is filtered, filtrate decompression is recycled to regulation crude drug amount, refrigerates,
Filtering.Then a certain amount of activated carbon is added, stirring certain time makes it fully adsorb, and filters.
It should be noted that being adsorbed to adding activated carbon in above-mentioned decoction, can be selected according to the situation of reality
Absorption is once or absorption is multiple.
Further, the pH for adjusting Radix Ophiopogonis extract is adjusted with inorganic alkali solution.
The basic skills and the step all same of extraction red ginseng of the tuber of dwarf lilyturf is extracted, specifically other steps refer to said extracted
The process of red ginseng.
S3, merge after using ultrafiltration membrance filter to red ginseng extract and Radix Ophiopogonis extract.
It is associated with using macromolecular substances in milipore filter removing decoction and easily the small molecular weight impurity of macromolecular substances.Can not only
Enough ensure that the Shenmai injection quality being prepared meets《Chinese Pharmacopoeia》Four injection item items Testing index, and do not have
Polyoxyethylene sorbitan monoleate auxiliary material is added, adverse reaction clinical safety hidden danger can greatly be reduced.
Further, ultrafiltration retaining molecular weight is 10KD~50KD.So as to effectively filter off except in decoction
The small molecular weight impurity of macromolecular substances.
It is further preferred that above-mentioned ultrafiltration retaining molecular weight is 30KD.The removal of impurity can not only be effectively gone, and
Integrated cost is relatively low.
A kind of it should be understood that aperture that above-mentioned Ultra filtration membrane, concentration, purifying biological product, pharmaceutical products field are common
Specification is consistent, and nominal pore scope is less than 0.01 micron of micropore filtering film.Other explanations should not be made herein.
Further, after merging red ginseng extract and Radix Ophiopogonis extract, add water to every 1 milliliter of decoction to decoction and given birth to containing red ginseng
Dose is 0.1g, and regulating liquid medicine pH value is to 7.5~8.5.
In an optional embodiment of the invention, specifically, above-mentioned red ginseng extract, Radix Ophiopogonis extract filtering are taken, is merged
Decoction, prescribed volume is supplied with water for injection, is stirred evenly, adjust pH value.Filtering, embedding, sterilizing, obtain Shenmai injection.
Some embodiments of the present invention also provide a kind of Shenmai injection, its by above-mentioned Shenmai injection preparation side
Method is prepared.Not only quality meets the Shenmai injection《Chinese Pharmacopoeia》Four injection item items Testing index, and do not have
There is addition polyoxyethylene sorbitan monoleate auxiliary material, so as to greatly reduce adverse reaction clinical safety hidden danger.
The feature and performance of the present invention are described in further detail with reference to embodiments：
Embodiment 1
A kind of Shenmai injection that the present embodiment provides, is prepared by following preparation method.
Got it filled material according to parts by weight：100 parts of red ginseng, 100 parts of the tuber of dwarf lilyturf.
Red ginseng extracts：Red ginseng is taken, shave, is added in refluxing extraction tank, is soaked with the ethanol of volumetric concentration 85% and the above
Bubble, backflow 3 times is reheated, 1 hour every time, each addition amount of alcohol was 2 times of red ginseng weight；Filter phegma, filtrate decompression
It is 0.3g/ml to be recycled to crude drug amount；Refrigeration, filtering.Activated carbon is added, stirring more than 30min makes it fully adsorb, filtered；Filter
Liquid adjusts pH to 5.5 with NaOH solution, is concentrated under reduced pressure into no alcohol taste, is 0.25g/ml with water for injection moisturizing to crude drug amount, stirs evenly,
PH value is adjusted to be refrigerated, filtration to 5.0 with NaOH solution；Filtrate is boiled into 30min, adding activated carbon and stirring 5 minutes makes it fully inhale
It is attached, filter, refrigeration is to be prepared.
The tuber of dwarf lilyturf extracts：Tuber of dwarf lilyturf medicinal material is taken, is crushed, is added in refluxing extraction tank, with the ethanol of volumetric concentration 85% and the above
Immersion, backflow 2 times is reheated, 1 hour every time, each addition amount of alcohol was 2 times of tuber of dwarf lilyturf weight；Phegma is filtered, filtrate subtracts
It is 0.3g/ml that pressure, which is recycled to crude drug amount,；Refrigeration, filtering.Activated carbon is added, stirring 30min makes it fully adsorb, filtered；Decompression
No alcohol taste is concentrated into, is 0.25g/ml with water for injection moisturizing to crude drug amount, stirs evenly, pH value is adjusted to 5.0 with NaOH solution, it is cold
Hide, filtration；Filtrate is boiled into 30min, adding activated carbon and stirring 5 minutes makes it fully adsorb, and filters, and refrigeration is to be prepared.
Above-mentioned red ginseng extract, Radix Ophiopogonis extract is taken to be filtered using ultrafiltration, ultrafiltration retaining molecular weight is 10KD.Merge
Decoction, moisturizing to every 1 milliliter of crude drug amount containing red ginseng is 0.1g, is stirred evenly, and adjusts pH value to 7.5.Filtering, embedding, sterilizing, wheat note must be joined
Penetrate liquid.
Embodiment 2
A kind of Shenmai injection that the present embodiment provides, is prepared by following preparation method.
Got it filled material according to parts by weight：100 parts of red ginseng, 100 parts of the tuber of dwarf lilyturf.
Red ginseng extracts：Red ginseng is taken, shave, is added in refluxing extraction tank, is soaked with the ethanol of volumetric concentration 85% and the above
Bubble, backflow 6 times is reheated, 2 hours every time, each addition amount of alcohol was 5 times of red ginseng weight；Filter phegma, filtrate decompression
It is 0.4g/ml to be recycled to crude drug amount；Refrigeration, filtering.Activated carbon is added, stirring more than 35min makes it fully adsorb, filtered；Filter
Liquid adjusts pH to 5.5 with NaOH solution, is concentrated under reduced pressure into no alcohol taste, is 0.50g/ml with water for injection moisturizing to crude drug amount, stirs evenly,
PH value is adjusted to be refrigerated, filtration to 8.0 with NaOH solution；Filtrate is boiled into 60min, the activated carbon of addition, which stirs 6 minutes, makes its abundant
Absorption, filter, refrigeration is to be prepared.
The tuber of dwarf lilyturf extracts：Tuber of dwarf lilyturf medicinal material is taken, is crushed, is added in refluxing extraction tank, with the ethanol of volumetric concentration 85% and the above
Immersion, backflow 3 times is reheated, 2 hours every time, each addition amount of alcohol was 5 times of tuber of dwarf lilyturf weight；Phegma is filtered, filtrate subtracts
It is 0.4g/ml that pressure, which is recycled to crude drug amount,；Refrigeration, filtering.Activated carbon is added, stirring 30min makes it fully adsorb, filtered；Decompression
No alcohol taste is concentrated into, is 0.50g/ml with water for injection moisturizing to crude drug amount, stirs evenly, pH value is adjusted to 8.0 with NaOH solution, it is cold
Hide, filtration；Filtrate is boiled into 60min, the activated carbon of addition, which stirs 5 minutes, makes it fully adsorb, and filters, and refrigeration is to be prepared.
Above-mentioned red ginseng extract, Radix Ophiopogonis extract is taken to be filtered using ultrafiltration, ultrafiltration retaining molecular weight is 50KD.Merge
Decoction, moisturizing to every 1 milliliter of crude drug amount containing red ginseng is 0.1g, is stirred evenly, and adjusts pH value to 8.5.Filtering, embedding, sterilizing, wheat note must be joined
Penetrate liquid.
Embodiment 3
A kind of Shenmai injection that the present embodiment provides, is prepared by following preparation method.
Got it filled material according to parts by weight：100 parts of red ginseng, 100 parts of the tuber of dwarf lilyturf.
Red ginseng extracts：Red ginseng is taken, shave, is added in refluxing extraction tank, is soaked with the ethanol of volumetric concentration 85% and the above
Bubble, backflow 5 times is reheated, 1.5 hours every time, each addition amount of alcohol was 4 times of red ginseng weight；Phegma is filtered, filtrate subtracts
It is 0.35g/ml that pressure, which is recycled to crude drug amount,；Refrigeration, filtering.Activated carbon is added, stirring more than 40min makes it fully adsorb, filtered；
Filtrate adjusts pH to 7.0 with NaOH solution, is concentrated under reduced pressure into no alcohol taste, is 0.3g/ml with water for injection moisturizing to crude drug amount, stirs
It is even, adjust pH value to be refrigerated, filtration to 7.0 with NaOH solution；Filtrate is boiled into 45min, adding activated carbon and stirring 7 minutes fills it
Divide absorption, filter, refrigeration is to be prepared.
The tuber of dwarf lilyturf extracts：Tuber of dwarf lilyturf medicinal material is taken, is crushed, is added in refluxing extraction tank, with the ethanol of volumetric concentration 85% and the above
Immersion, backflow 2 times is reheated, 1.5 hours every time, each addition amount of alcohol was 4 times of tuber of dwarf lilyturf weight；Filter phegma, filtrate
It is 0.35g/ml to be recovered under reduced pressure to crude drug amount；Refrigeration, filtering.Activated carbon is added, stirring 30min makes it fully adsorb, filtered；Subtract
Pressure is concentrated into no alcohol taste, is 0.30g/ml with water for injection moisturizing to crude drug amount, stirs evenly, and pH value is adjusted to 7.0 with NaOH solution, cold
Hide, filtration；Filtrate is boiled into 45min, adding activated carbon and stirring 6 minutes makes it fully adsorb, and filters, and refrigeration is to be prepared.
Above-mentioned red ginseng extract, Radix Ophiopogonis extract is taken to be filtered using ultrafiltration, ultrafiltration retaining molecular weight is 30KD.Merge
Decoction, moisturizing to every 1 milliliter of crude drug amount containing red ginseng is 0.1g, is stirred evenly, and adjusts pH value to 8.0.Filtering, embedding, sterilizing, wheat note must be joined
Penetrate liquid.
Embodiment 4
A kind of Shenmai injection that the present embodiment provides, is prepared by following preparation method.
Got it filled material according to parts by weight：100 parts of red ginseng, 100 parts of the tuber of dwarf lilyturf.
Red ginseng extracts：Red ginseng is taken, shave, adds in refluxing extraction tank, is soaked with methanol, reheats backflow 5 times, every time
2 hours, each addition quantity of methyl alcohol was 3.5 times of red ginseng weight；Phegma is filtered, it is 0.35g/ that filtrate decompression, which is recycled to crude drug amount,
ml；Refrigeration, filtering.Activated carbon is added, stirring more than 40min makes it fully adsorb, filtered；Filtrate adjusts pH extremely with NaOH solution
7.0, no alcohol taste is concentrated under reduced pressure into, is 0.3g/ml with water for injection moisturizing to crude drug amount, stirs evenly, pH value is adjusted extremely with NaOH solution
7.0, refrigerate, filtration；Filtrate is boiled into 45min, adding activated carbon and stirring 7 minutes makes it fully adsorb, and filters, and refrigeration, waits to match somebody with somebody
System.
The tuber of dwarf lilyturf extracts：Tuber of dwarf lilyturf medicinal material is taken, is crushed, adds in refluxing extraction tank, is soaked with ethyl acetate, reheats backflow 2
Secondary, 2 hours every time, each addition ethyl acetate amount was 4 times of tuber of dwarf lilyturf weight；Phegma is filtered, filtrate decompression is recycled to crude drug
Measure as 0.35g/ml；Refrigeration, filtering.Activated carbon is added, stirring 30min makes it fully adsorb, filtered；It is concentrated under reduced pressure into no alcohol
Taste, it is 0.30g/ml with water for injection moisturizing to crude drug amount, stirs evenly, adjusts pH value to be refrigerated, filtration to 7.0 with NaOH solution；Will filter
Liquid boils 45min, and adding activated carbon and stirring 6 minutes makes it fully adsorb, and filters, and refrigeration is to be prepared.
Above-mentioned red ginseng extract, Radix Ophiopogonis extract is taken to be filtered using ultrafiltration, ultrafiltration retaining molecular weight is 30KD.Merge
Decoction, moisturizing to every 1 milliliter of crude drug amount containing red ginseng is 0.1g, is stirred evenly, and adjusts pH value to 8.0.Filtering, embedding, sterilizing, wheat note must be joined
Penetrate liquid.
Experimental example
The embodiment 1-4 Shenmai injections provided are detected, detect whether it contains polyoxyethylene sorbitan monoleate.
1st, the Shenmai injection that embodiment 1-4 is provided is accelerated 6 months and 25 DEG C to place 36 months at 40 DEG C, decoction clarification
Degree is good, meets《Pharmacopoeia of People's Republic of China》Testing requirements under injection item.
2nd, detected using HPLC methods in the Shenmai injection that embodiment 1-4 provides without oleic acid；Above-mentioned physicochemical characteristicses
Testing conditions are as follows：Chromatographic column (2.0mm × 150mm, 3 μm) of the stationary phase using octadecylsilane chemically bonded silica as filler, with
Acetonitrile is mobile phase A, and 0.01% phosphoric acid is Mobile phase B, and from 0~50 minute, Mobile phase B was uniformly changed into 0% from 70%；Flow velocity is
0.4ml/min, keep 40 DEG C of column temperature, Detection wavelength 203nm.It is prepared by reference substance solution：Take oleic acid reference substance appropriate, precision claims
It is fixed, add acetonitrile that the solution that every 1ml contains 2 μ g is made, produce.Assay：Precision draws this product and reference substance solution, is injected separately into
In high performance liquid chromatography, it is measured according to above-mentioned chromatographic condition.
3rd, detected using HPLC-GPC-ELSD methods in the Shenmai injection that embodiment 1-4 provides without polyoxyethylene sorbitan monoleate；On
The testing conditions for stating physicochemical characteristicses are as follows：Stationary phase use hydrophilic silica gels for filler (TSK GEL G2000SWxl 7.8mm ×
300mm, 5 μm) chromatographic column；With acetonitrile -20mmol/L ammonium acetates (10:90) it is mobile phase；Flow velocity is 0.6ml per minute；Evaporation
Light scattering detector detects.It is prepared by reference substance solution：Take polyoxyethylene sorbitan monoleate reference substance appropriate, it is accurately weighed, add water that every 1ml is made
Solution containing 2.0mg/ml, is produced.Assay：Precision draws this product and reference substance solution, is injected separately into high performance liquid chromatography
In, it is measured according to above-mentioned chromatographic condition.
Fig. 1 is refer to, Ginsenosides finger-print is detected using HPLC methods, with ginsenoside Rb1's peak area
5% integration, a total of 23 characteristic peaks, wherein there have 11 characteristic peaks to point out to be as follows：Peak #2 peaks are ginsenoside Re, #3 peaks are
Ginsenoside Rg1, #4 peaks are ginsenoside Rf, #5 peaks are ginsenoside Rb1, #6 peaks are ginsenoside Rg2, and #7 peaks are ginseng soap
Glycosides Rc, #8 peaks are ginsenoside Rh 1, and #9 peaks are ginsenoside Rb2, and #10 peaks are ginsenoside Rb3, and #12 peaks are ginsenoside
Rd, #22 peak are ginsenoside Ck.
In finger-print, it is respectively by the relative retention time with reference to peak, 23 characteristic peaks of ginsenoside Rb1：0.30
~0.35 (#1), 0.32~0.37 (#2), 0.34~0.39 (#3), 0.82~0.87 (#4), 1.000 (#5), 1.05~1.10
(#6), 1.08~1.13 (#7), 1.10~1.15 (#8), 1.13~1.18 (#9), 1.14~1.19 (#10), 1.18~1.23
(#11), 1.23~1.27 (#12), 1.40~1.45 (#13), 1.45~1.50 (#14), 1.48~1.53 (#15), 1.52~
1.57 (#16), 1.54~1.59 (#17), 1.59~1.64 (#18), 1.61~1.66 (#19), 1.78~1.83 (#20),
1.80~1.85 (#21), 1.82~1.87 (#22), 1.87~1.92 (#23)；Using ginsenoside Rb1 as with reference to peak, 23 spies
Sign peak relative peak area be respectively：0.10~1.5 (#1), 0.40~0.45 (#2), 0.8~0.85 (#3), 0.27~0.32
(#4), 1.000 (#5), 0.07~0.12 (#6), 0.55~0.60 (#7), 0.17~0.22 (#8), 0.40~0.45 (#9),
0.15~0.20 (#10), 0.05~0.10 (#11), 0.25~0.30 (#12), 0.05~0.10 (#13), 0.05~0.10 (#
14), 0.10~0.15 (#15), 0.09~0.14 (#16), 0.42~0.47 (#17), 0.22~0.27 (#18), 0.15~
0.20 (#19), 0.16~0.21 (#20), 0.27~0.32 (#21), 0.56~0.61 (#22), 0.23~0.28 (#23).On
The condition determination for stating physical and chemical parameter is as follows：Chromatographic condition：Stationary phase is with the chromatographic column of octadecylsilane chemically bonded silica filler
(4.6mm × 250mm, 5.0 μm)；Elution requirement：Acetonitrile is mobile phase A, and water is Mobile phase B, from 0~10 minute, Mobile phase B from
72% is uniformly changed into 66%, and from 10~15 minutes, Mobile phase B was uniformly changed into 64% from 66%, from 15~25 minutes, Mobile phase B
Uniformly it is changed into 40% from 64%, from 25~30 minutes, Mobile phase B was uniformly changed into 20% from 40%, from 30~35 minutes, mobile phase
B is uniformly changed into 0% from 20%.Flow velocity is 1.0ml/min, and it is 40~50 DEG C to keep column temperature, and Detection wavelength is 200~220nm.Ginseng
According to the preparation of thing solution：It is appropriate that precision weighs ginsenoside Rb1's reference substance, adds 20% acetonitrile that every 1ml is made and contains 0.20mg respectively
Solution, produce.Determining fingerprint pattern：Precision draws Shenmai injection finished product and each 10 μ l of reference solution, is injected separately into height
Effect liquid phase chromatogram instrument, it is measured according to above-mentioned chromatographic condition, obtains high-efficient liquid phase chromatogram.
In finger-print, it is respectively by the relative retention time with reference to peak, 23 characteristic peaks of ginsenoside Rb1：0.323
(#1)、0.348(#2)、0.363(#3)、0.858(#4)、1.000(#5)、1.070(#6)、1.096(#7)、1.111(#8)、
1.151(#9)、1.162(#10)、1.197(#11)、1.256(#12)、1.422(#13)、1.478(#14)、1.508(#15)、
1.542(#16)、1.577(#17)、1.611(#18)、1.632(#19)、1.805(#20)、1.824(#21)、1.843(#22)、
1.893(#23)；It is respectively by the relative peak area with reference to peak, 23 characteristic peaks of ginsenoside Rb1：0.113(#1)、0.434
(#2)、0.820(#3)、0.296(#4)、1.000(#5)、0.086(#6)、0.572(#7)、0.197(#8)、0.418(#9)、
0.167(#10)、0.072(#11)、0.264(#12)、0.073(#13)、0.079(#14)、0.130(#15)、0.114(#16)、
0.449(#17)、0.241(#18)、0.173(#19)、0.186(#20)、0.292(#21)、0.584(#22)、0.252(#23)。
The condition determination of above-mentioned physical and chemical parameter is as follows：Chromatographic condition：Stationary phase is with the chromatographic column of octadecylsilane chemically bonded silica filler
(4.6mm × 250mm, 5.0 μm)；Elution requirement：Acetonitrile is mobile phase A, and water is Mobile phase B, from 0~10 minute, Mobile phase B from
72% is uniformly changed into 66%, and from 10~15 minutes, Mobile phase B was uniformly changed into 64% from 66%, from 15~25 minutes, Mobile phase B
Uniformly it is changed into 40% from 64%, from 25~30 minutes, Mobile phase B was uniformly changed into 20% from 40%, from 30~35 minutes, mobile phase
B is uniformly changed into 0% from 20%.Flow velocity is 1.0ml/min, and it is 40~50 DEG C to keep column temperature, and Detection wavelength is 200~220nm.Ginseng
According to the preparation of thing solution：It is appropriate that precision weighs ginsenoside Rb1's reference substance, adds 20% acetonitrile that every 1ml is made and contains 0.20mg respectively
Solution, produce.Determining fingerprint pattern：Precision draws Shenmai injection finished product and each 10 μ l of reference solution, is injected separately into height
Effect liquid phase chromatogram instrument, it is measured according to above-mentioned chromatographic condition, obtains high-efficient liquid phase chromatogram.
To sum up, the Shenmai injection that embodiment 1-4 is provided, does not detect polyoxyethylene sorbitan monoleate, meets《Chinese Pharmacopoeia》
Injection general rule items Testing index.
Comparative example 1
A kind of Shenmai injection provided, add 0.25% polyoxyethylene sorbitan monoleate.
Comparative example 2
A kind of Shenmai injection provided, add 0.5% polyoxyethylene sorbitan monoleate.
It is the anaphylactoid reality of Shenmai injection cavy that the present invention is provided embodiment 1-4 and comparative example 1-2 below
Test pharmacological research.
1 materials and methods
1.1 animal cavys 40, male and female half and half, female body weight 300-400g, it is special to be purchased from Sichuan Province experimental animal without pregnancy
Wei Hui plants, animal credit number：SCXK (river) 2013-14.Feed is purchased from plant of experimental animal special commission of Sichuan Province, is permitted
Can the number of card：SCXK (river) 2013-14.
1.2 medicines and three kinds of Shenmai injections of reagent (polyoxyethylene sorbitan monoleate addition is respectively 0%, 0.25%, 0.5%).
0.9% sodium chloride solution (is made into the sodium chloride solution that concentration is 0.9% with appropriate sodium chloride, water for injection；Filling 10ml/ branch,
Sealing；121 DEG C, sterilize within 12 minutes, produce).Egg albumin (freeze-dried powder) (Sigma companies).The Guinea pig IgE (U.S.
Rapidbio lab).Guinea pig Histamine (U.S. Rapidbio Lab).
1.3 hole instrument high-speed refrigerated centrifuges, model IEC-CL3IR, Thermo Fisher Scientific Inc.；ELIASA, type
Number MultisKan FC, Thermo Fisher Scientific Inc.；Electro-heating standing-temperature cultivator, BPH-9052, the allusion quotation instrument of Shanghai City hundred
Factory.
Cavy is randomly divided into 5 groups, every group 8 by 1.4 animal packets.A groups：Negative control group (0.9% chloride injection
Liquid)；B groups：Positive controls (0.15mg/ml ovalbumins liquid)；C groups：Shenmai injection control group (C1 groups：Containing 0% poly- sorb
The Shenmai injection group of ester 80；C2 groups：Containing 0.25% polyoxyethylene sorbitan monoleate Shenmai injection group；C3 groups：Join containing 0.5% polyoxyethylene sorbitan monoleate
Wheat parenteral solution group).
1.5 test methods use intraperitoneal injection sensitization, administered volume be 1ml/ only, the next day once, totally 4 times.Last causes
11st day after quick, each group cavy is excited by digital veins of the foot injection respective sets decoction, and booster dose is 2ml/.During sensitization
Daily observation animal state, intravenous injection excite Continuous Observation cavy reaction 40min after preceding 15min and injection, and record symptom goes out
Between current, extinction time.Blood is taken after exciting drug withdrawal 40min, anticoagulant heparin, prepares blood plasma.
1.6 observation index
1.6.1 during reaction of animals symptom sensitization and when exciting administration, observe and record the reaction symptom of animal and continue
Time, such as scratch nose, sneeze, it is unpeaceful, jump, pant, purpura, and according to whole body sensitization reaction symptom grade scale (table
1) scoring and the rank of reaction symptom are judged.
The whole body sensitization reaction symptom grade scale of table 1
1.6.2 biochemical indicator is using IGE, histamine content in double-antibody method ELISA kit detection blood plasma.And according to anti-
Answer symptom rank and IGE, histamine elevated-levels synthetic determination sensitization standard (table 2), synthetic determination sensitization.
The synthetic determination sensitization standard of table 2
| Reaction symptom rank | IGE contents elevated-levels/% | Histamine content elevated-levels/% | Result of determination |
| It is negative | <30 | <30 | It is negative |
| Weakly positive | 30~100 | 30~100 | It is suspicious |
| It is positive and more than | >100 | >100 | It is positive |
Note：Reaction of animals symptom, IgE, histamine content are raised 3 indexs reach delimitation line simultaneously when to be judged by table 2, if
3 indexs do not reach simultaneously, but have significant difference to be, using high target as criterion, a rank is dropped during judgement.
1.7 statistical method measurement datas are using " ± s " represents that the comparison between each group is using single factor test variance point
Analysis, homogeneity of variance are examined with SNK, and heterogeneity of variance Tamhane ' s T2 are examined.P<0.05 is statistically significant.
2 result of the tests
2.1 cavy reaction symptom each group animals do not occur abnormal symptom in sensitization first.Diet, drink during sensitization
Water, behavior are showed no exception, and Normal-weight increases.A groups animal does not occur allergic reaction after administration is excited；B groups excite to
Occur strong allergic reaction after medicine, show as spasm, purpura, instability of gait, pant, jump, shedding tears, be One's spirits are drooping, 7 are being swashed
Issue dead in 2min after medicine；C1 groups animal does not occur allergic reaction after administration is excited；C2 groups have 2 cavys to scratch
Grab, unpeaceful symptom, remaining cavy is without exception；There is jump, instability of gait symptom by 1 cavy in C3 groups, and 2 cavys occur
There is unpeaceful, perpendicular hair, shake, scratch nasal symptom in urination, defecation symptom, 3 cavys, and remaining 2 cavy is without exception.According to table
1 judges each group cavy reaction symptom rank, and B groups are the extremely strong positive, and C3 groups are the positive, and C2 groups are weakly positive, and A groups, C1 groups are the moon
Property.Reaction symptom classification results are shown in Table 3 after each group cavy excites administration.
The each group cavy of table 3 excites reaction symptom classification results (n=8) after administration
After 2.2 blood plasma IGE and histamine content each group animal excite administration, blood plasma IGE contents are above A groups, but without notable
Sex differernce (P>0.05), elevated-levels are below 100%.Plasma Histamine content is above A groups, wherein B groups histamine content and A groups
Comparing has significant difference (P<0.05)；C1, C2, C3 group Plasma Histamine content average value are higher than A groups, but there was no significant difference (P
>0.05) 4, are shown in Table.
The each group cavy of table 4 excites after administration IGE and histamine content (n=8, ± s) in blood plasma
Note：Rate of change (%)=(each class value-A class values)/A class value × 100%
2.3 synthetic determination sensitization combination each group animals excite the reaction symptom occurred after administration to become with blood plasma IGE, histamine
Change situation, sensitization of the tested decoction of synthetic determination each group to cavy.As a result show, B groups are the positive, and C1, C2 group is negative, C3
Group is suspicious.
The each group of table 5 is by test solution to cavy sensitization synthetic determination result (n=8)
This research is positive control and using 0.9% sodium chloride injection as negative control using egg albumin, be have detected not
With the Shenmai injection finished product of polyoxyethylene sorbitan monoleate addition, as a result find as the increase of polyoxyethylene sorbitan monoleate addition, cavy go out
Existing allergic reaction number of elements increase；Detection serum I GE and histamine content, its average also with the increase of polyoxyethylene sorbitan monoleate addition, go out
Now increase trend；Comprehensive three Testing index are thought to be proportionate with allergic reaction in polyoxyethylene sorbitan monoleate addition；Addition is low
In 0.25% safety of Shenmai injection be higher than the addition 0.5% of tween 90 Shenmai injection.
Therefore, paradoxical reaction is no different during each group cavy sensitization, excites rear synthetic determination sensitization；Ovalbumin positive control
Group induces cavy strong positive allergic reaction；0.5% polyoxyethylene sorbitan monoleate Shenmai injection group induces cavy positive allergic reaction,
0.25% polyoxyethylene sorbitan monoleate Shenmai injection group induces cavy weakly positive allergic reaction, and other each groups do not induce cavy and allusion quotation occur
The allergic reaction of type；Each group blood plasma IgE contents are above negative control group content, but there was no significant difference (P>0.05)；Each group
Plasma Histamine content is above negative control group content, and wherein positive controls have significant difference (P with negative control group<
0.05), the not notable (P of sample sets difference>0.05).
In summary, reduce polyoxyethylene sorbitan monoleate addition in Shenmai injection (being less than 0.25%) or be added without polysorbate
80, cavy allergic reaction generation can be reduced.
It should be understood that although being illustrated with specific embodiment and being described the present invention, but will be appreciated that without departing substantially from this hair
Many other change and modification can be made in the case of bright spirit and scope.It is, therefore, intended that will in appended right
Ask including belonging to all such changes and modifications in the scope of the invention.
Claims (10)
1. a kind of preparation method of Shenmai injection, it is characterised in that the Shenmai injection is made up of red ginseng, the tuber of dwarf lilyturf and water；
The preparation method of the Shenmai injection comprises the following steps：
Red ginseng is extracted with the first extractant, the solvent in the first extract solution is removed, adds water, obtains red ginseng extract, regulation is described red
Join the pH to 5.0~8.0 of extract solution；
The tuber of dwarf lilyturf is extracted with the second extractant, the solvent in the second extract solution is removed, adds water, obtain Radix Ophiopogonis extract, adjust the wheat
The pH of winter extract solution to 5.0~8.0；
Merge after using ultrafiltration membrance filter to the red ginseng extract and the Radix Ophiopogonis extract；
First extractant and second extractant are organic solvent；The organic solvent is selected from methanol, ethanol and second
Any one in acetoacetic ester.
2. the preparation method of Shenmai injection according to claim 1, it is characterised in that
Adjust the pH to 7.0 of the red ginseng extract；
Adjust the pH to 8.0 of the Radix Ophiopogonis extract.
3. the preparation method of Shenmai injection according to claim 1, it is characterised in that
It is red ginseng 3~6 times described in the first extractant heating and refluxing extraction to extract red ginseng；
It is the tuber of dwarf lilyturf 2~3 times described in the second extractant heating and refluxing extraction to extract the tuber of dwarf lilyturf.
4. the preparation method of Shenmai injection according to claim 1, it is characterised in that
The weight ratio of first extractant and the red ginseng is 2~5：1；
Second extractant and the weight ratio of the tuber of dwarf lilyturf are 2~5：1.
5. the preparation method of Shenmai injection according to claim 1, it is characterised in that remove in first extract solution
Solvent and be additionally added activated carbon when removing the solvent in second extract solution and adsorbed.
6. the preparation method of Shenmai injection according to claim 1, it is characterised in that
Adjusting the pH of the red ginseng extract and the pH of the regulation Radix Ophiopogonis extract is adjusted with inorganic alkali solution.
7. the preparation method of the Shenmai injection according to claim any one of 1-6, it is characterised in that
The ultrafiltration retaining molecular weight is 10KD~50KD.
8. the preparation method of Shenmai injection according to claim 7, it is characterised in that
The ultrafiltration retaining molecular weight is 30KD.
9. the preparation method of Shenmai injection according to claim 8, it is characterised in that
After merging the red ginseng extract and the Radix Ophiopogonis extract, add water to every 1 milliliter of decoction to decoction and given birth to containing red ginseng
Dose is 0.1g, and adjusts the decoction pH value to 7.5~8.5.
A kind of 10. Shenmai injection, it is characterised in that
It is prepared as the preparation method of the Shenmai injection described in claim any one of 1-9.
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| CN101375990A (en) * | 2007-08-29 | 2009-03-04 | 济南天瑞本草医药科技有限公司 | Composition for treating cardiovascular disease and preparation thereof |
| CN101623437A (en) * | 2009-08-04 | 2010-01-13 | 江苏苏中药业集团股份有限公司 | Pulse activating preparation for injection and its preparation method |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101375990A (en) * | 2007-08-29 | 2009-03-04 | 济南天瑞本草医药科技有限公司 | Composition for treating cardiovascular disease and preparation thereof |
| CN101623437A (en) * | 2009-08-04 | 2010-01-13 | 江苏苏中药业集团股份有限公司 | Pulse activating preparation for injection and its preparation method |
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