Summary of the invention
The objective of the invention is to provide the preparation method of acanthopanax senticosus extract/thing at above-mentioned deficiency.
Preparation method of the present invention comprises the steps: to adopt decoction and alcohol sedimentation technique to prepare the Radix Et Caulis Acanthopanacis Senticosi drug extract; With the extractum thin up, with lime cream adjusting pH to 10.00~12.00, pH to 4.5~6.5 are regulated with sulphuric acid in the back that stirs, and leave standstill, and extract supernatant liquid filtering, concentrate, and precipitate with ethanol, remove impurity is left standstill in cold preservation, gets acanthopanax senticosus extract; Or concentrated drying under reduced pressure gets Radix Et Caulis Acanthopanacis Senticosi extract.
The present invention utilizes lime cream to transfer alkali, makes total flavonoid form the sequestration thing with it, adds the sulphuric acid acid adjustment again, and the total flavonoid composition is dissolved in the solution, and the impurity component precipitation, this method has increased the stripping of total flavones, and can more effectively remove impurity.
More specifically, extractum is diluted with water to 15 times more than the volume, stirs, treat that temperature reduces to below 50 ℃, add the fresh lime cream of 10-30%, adjust pH to 10.00~12.00 are after stirring, reuse 10-30% sulphuric acid adjust pH to 4.5~6.5, continue to stir after 5~10 minutes, be cooled to room temperature, left standstill 4~6 hours, extract supernatant liquid filtering, further improve the effect of remove impurity again through precipitate with ethanol.Here the method for precipitate with ethanol is, filtrate decompression is concentrated into relative density 1.08~1.17 (80 ℃ of surveys), when treating that temperature is reduced to below 40 ℃, slowly adds ethanol and makes and contain alcohol amount and reach 82~85%, precipitates 12~36 hours.
The method that above-mentioned decoction and alcohol sedimentation technique prepares drug extract is: the Radix Et Caulis Acanthopanacis Senticosi bar is cut into pieces, add water to few extraction 2 times, add 2~10 times of water gagings at every turn, decocted 30~60 minutes, merging filtrate concentrates, and 2 precipitate with ethanol leave standstill, filter, reclaim ethanol, promptly get drug extract to there not being the alcohol flavor.
More specifically, can prepare extractum by following method: get the Radix Et Caulis Acanthopanacis Senticosi bar, cut into the fragment of 1~3cm, decoct 2~4 times, add 4~8 times of water gagings at every turn, decocted 30~60 minutes, collecting decoction is concentrated into relative density 1.08~1.17 (80 ℃ of surveys), when treating that temperature is reduced to below 40 ℃, adding ethanol makes and contains the alcohol amount and reach 65~75%, left standstill after stirring 6-18 hour, and filtered, residue adds ethanol to be made and contains the alcohol amount and reach 75~85%, left standstill 4~12 hours, filter, filtrate is collected in residue centrifugal filtration, recovery ethanol gets equivalent extract to there not being the alcohol flavor.
The present invention carries out the purpose that cold preservation leaves standstill and is further remove impurity, the cold preservation standing methods is: get the precipitate with ethanol supernatant liquid filtering, residue centrifugal filtration, reclaim ethanol to not having the alcohol flavor and being concentrated into relative density 1.05~1.20 (80 ℃ of surveys), add water for injection and be diluted to 5g (crude drug)/ml, sterilization, freezing, negative catalysis, cold preservation is filtered.
More specifically, can leave standstill remove impurity by following method cold preservation: get supernatant liquid filtering, residue centrifugal filtration, reclaiming ethanol does not extremely have the alcohol flavor and is concentrated into relative density 1.08~1.17 (80 ℃ of surveys), adds water for injection and is diluted to 5g (crude drug)/ml, sterilizes 40 minutes for 115 ℃, freezing more than 12 hours down in-20 ℃~-10 ℃, negative catalysis, cold preservation are more than seven days, and medicinal liquid is through 0.45 μ m membrane filtration.
Preparation technology's advantage of the present invention: the content height of total flavones and syringoside, impurity is few, and technology is simple, and rationally, cost is low, pollutes few.
1) precipitate with ethanol repeatedly repeatedly can effectively be removed impurity such as macro-molecular protein.
2) add lime cream earlier and transfer alkali, make the total flavonoid composition form the sequestration thing with it, add the sulphuric acid acid adjustment again, the total flavonoid composition is dissolved in the solution, and the impurity component precipitation has increased the stripping of total flavones, and can more effective removal impurity.
Acanthopanax senticosus extract/the thing and the pharmaceutic adjuvant that can further method for preparing be obtained are prepared into various dosage forms, comprise injection, lyophilized injectable powder, infusion solution, oral liquid, capsule, granule and tablet etc.Described pharmaceutic adjuvant comprises glucose, ethanol (95%), mannitol, dextran, lactose, sodium chloride, polyoxyethylene sorbitan monoleate etc.When said preparation was injection, it was 1.8-5.5mg that this injection of every 1ml contains total flavones; Contain syringoside and should be 0.20-2.0mg.
The specific embodiment
Further set forth the present invention below in conjunction with specific embodiment.Should be appreciated that these embodiment only are used to illustrate the present invention, and can not limit protection scope of the present invention.
The preparation of embodiment 1 acanthopanax senticosus extract
Get the Radix Et Caulis Acanthopanacis Senticosi bar, cut into the fragment of 1~3cm.Decoct three times, add 5 times of water gagings for the first time and decocted 30 minutes; Adding for the second time 6 times of water gagings decocted 40 minutes; Adding 10 times of water gagings for the third time decocted 30 minutes.Collecting decoction is concentrated into relative density 1.10 (80 ℃ of surveys), when treating that temperature is reduced to below 40 ℃, adding ethanol makes and contains the alcohol amount and reach 70%, left standstill after stirring 10 hours, and filtered, residue adds ethanol to be made and contains the alcohol amount and reach 80%, left standstill 8 hours, filter, filtrate is collected in residue centrifugal filtration, recovery ethanol gets equivalent extract to there not being the alcohol flavor.Add purified water to 15 times of volumes of equivalent extract, stir, treat that temperature reduces to below 50 ℃, add 20% fresh lime cream, adjust pH to 11.00 is after stirring, reuse 20% sulphuric acid adjust pH to 5.5, continue to stir after 8 minutes, be cooled to room temperature, left standstill 5 hours, extract supernatant liquid filtering, be evaporated to relative density 1.15 (80 ℃ of surveys), when treating that temperature is reduced to below 40 ℃, slowly add ethanol and make and contain alcohol amount and reach 85%, precipitate 24 hours, get supernatant liquid filtering, residue centrifugal filtration is reclaimed ethanol to not having the alcohol flavor and being concentrated into relative density 1.05 (80 ℃ of surveys).Add water for injection and be diluted to 5g (crude drug)/ml, sterilized 40 minutes for 115 ℃.Descended negative catalysis, cold preservation seven days freezing 12 hours in-10 ℃.Medicinal liquid promptly gets acanthopanax senticosus extract through 0.45 μ m membrane filtration.
The preparation of embodiment 2 Radix Et Caulis Acanthopanacis Senticosi extracts
Get the Radix Et Caulis Acanthopanacis Senticosi bar, cut into the fragment of 1~3cm.Decoct 2 times, add 4 times of water gagings for the first time and decocted 40 minutes; Adding for the second time 8 times of water gagings decocted 60 minutes.Collecting decoction is concentrated into relative density 1.08 (80 ℃ of surveys), when treating that temperature is reduced to below 40 ℃, adding ethanol makes and contains the alcohol amount and reach 75%, left standstill after stirring 18 hours, and filtered, residue adds ethanol to be made and contains the alcohol amount and reach 85%, left standstill 12 hours, filter, filtrate is collected in residue centrifugal filtration, recovery ethanol gets equivalent extract to there not being the alcohol flavor.Add purified water to 18 times of volumes of equivalent extract, stir, treat that temperature reduces to below 50 ℃, add 10% fresh lime cream, adjust pH to 12.00 is after stirring, reuse 10% sulphuric acid adjust pH to 6.5, continue to stir after 10 minutes, be cooled to room temperature, left standstill 6 hours, extract supernatant liquid filtering, be evaporated to relative density 1.17 (80 ℃ of surveys), when treating that temperature is reduced to below 40 ℃, slowly add ethanol and make and contain alcohol amount and reach 82%, precipitate 36 hours, get supernatant liquid filtering, residue centrifugal filtration is reclaimed ethanol to not having the alcohol flavor and being concentrated into relative density 1.08 (80 ℃ of surveys).Add water for injection and be diluted to 5g (crude drug)/ml, sterilized 40 minutes for 115 ℃.Descended negative catalysis, cold preservation ten days freezing 16 hours in-15 ℃.Medicinal liquid is through 0.45 μ m membrane filtration, and concentrated drying under reduced pressure promptly gets Radix Et Caulis Acanthopanacis Senticosi extract.
The preparation of embodiment 3 acanthopanax senticosus extracts
Get the Radix Et Caulis Acanthopanacis Senticosi bar, cut into the fragment of 1~3cm.Decoct 3 times, add 5 times of water gagings for the first time and decocted 30 minutes; Adding for the second time 6 times of water gagings decocted 40 minutes; Adding 4 times of water gagings for the third time decocted 50 minutes.Collecting decoction is concentrated into relative density 1.17 (80 ℃ of surveys), when treating that temperature is reduced to below 40 ℃, adding ethanol makes and contains the alcohol amount and reach 65%, left standstill after stirring 6 hours, and filtered, residue adds ethanol to be made and contains the alcohol amount and reach 75%, left standstill 4 hours, filter, filtrate is collected in residue centrifugal filtration, recovery ethanol gets equivalent extract to there not being the alcohol flavor.Add purified water to 15 times of volumes of equivalent extract, stir, treat that temperature reduces to below 50 ℃, add 30% fresh lime cream, adjust pH to 10.00 is after stirring, reuse 30% sulphuric acid adjust pH to 4.5, continue to stir after 5 minutes, be cooled to room temperature, left standstill 4 hours, extract supernatant liquid filtering, be evaporated to relative density 1.17 (80 ℃ of surveys), when treating that temperature is reduced to below 40 ℃, slowly add ethanol and make and contain alcohol amount and reach 84%, precipitate 12 hours, get supernatant liquid filtering, residue centrifugal filtration is reclaimed ethanol to not having the alcohol flavor and being concentrated into relative density 1.17 (80 ℃ of surveys).Add water for injection and be diluted to 5g (crude drug)/ml, sterilized 40 minutes for 115 ℃.Descended negative catalysis, cold preservation 15 days freezing 20 hours in-20 ℃.Medicinal liquid promptly gets acanthopanax senticosus extract through 0.45 μ m membrane filtration.
The preparation of embodiment 4 injection
Get the extracting solution of embodiment 1, with 40% NaOH solution adjust pH to 5.5, post-heating stirs, the active carbon of adding 0.2%, boil absorption 15 minutes, be cooled to below 50 ℃ NaOH solution adjust pH to 5.5, add fresh water for injection to original volume with 40%.Through 0.45 μ m membrane filtration to clarification, fill, 115 ℃ of autoclavings 40 minutes.Freezing, cold preservation is 15 days after the negative catalysis, and room temperature was placed 3 months.0.65 μ m filter element filtering adds the injection water to original volume, adjust pH to 5.5 adds activated carbon decolorizing, filters, and adjust pH to 5.5 is added water for injection to original volume, through 0.45 μ m, 0.22 μ m filter element fine straining, and embedding, 115 ℃ of sterilizations in 30 minutes, promptly.
Through check, the every 1ml of this injection contains total flavones, with anhydrous rutin (C
27H
30O
16) count 2.0mg, contain syringoside 0.20mg.
The preparation of embodiment 5 injection
Get the extract of embodiment 2, add the injection water and make dissolving, NaOH solution adjust pH to 5.5 with 40%, post-heating stirs, the active carbon of adding 0.2%, boil absorption 15 minutes, be cooled to below 50 ℃ NaOH solution adjust pH to 5.7, add fresh water for injection to containing crude drug 5g/ml with 40%.Through 0.45 μ m membrane filtration to clarification, fill, 115 ℃ of autoclavings 40 minutes.Freezing, cold preservation is 20 days after the negative catalysis, and room temperature was placed 3.5 months.0.65 μ m filter element filtering adds the injection water to containing crude drug 5g/ml, adjust pH to 5.0 adds activated carbon decolorizing, filter, adjust pH to 5.0 is added water for injection to containing crude drug 5g/ml, respectively through 0.45 μ m, 0.22 μ m filter element fine straining, embedding, 115 ℃ of sterilizations in 30 minutes, promptly.
Through check, the every 1ml of this injection contains total flavones, with anhydrous rutin (C
27H
30O
16) count 5.1mg, contain syringoside 2.0mg.
The preparation of embodiment 6 injection
Get the extracting solution of embodiment 3, with 40% NaOH solution adjust pH to 5.5, post-heating stirs, the active carbon of adding 0.2%, boil absorption 15 minutes, be cooled to below 50 ℃ NaOH solution adjust pH to 5.3, add fresh water for injection to original volume with 40%.Through 0.45 μ m membrane filtration to clarification, fill, 115 ℃ of autoclavings 40 minutes.Freezing, cold preservation is 25 days after the negative catalysis, and room temperature was placed 4 months.0.65 μ m filter element filtering adds the injection water to original volume, adjust pH to 6.0 adds activated carbon decolorizing, filters, and adjust pH to 6.0 is added water for injection to original volume, respectively through 0.45 μ m, 0.22 μ m filter element fine straining, and embedding, 115 ℃ of sterilizations in 30 minutes, promptly.
Through check, the every 1ml of this injection contains total flavones, with anhydrous rutin (C
27H
30O
16) count 3.3mg, contain syringoside 0.85mg.