CN103202884B - Method for extracting hovenaia dulcis total alkaloid by semi-bionic-enzymic method - Google Patents
Method for extracting hovenaia dulcis total alkaloid by semi-bionic-enzymic method Download PDFInfo
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Abstract
The invention discloses a method for extracting hovenaia dulcis total alkaloids by a semi-bionic-enzymic method and belongs to the technical field of alkaloid extraction. The method comprises the following steps of: 1) taking the hovenaia dulcis, adding a disodium hydrogen phosphate-citric acid buffer solution for regulating the pH value to the range from 1.5 to 3.0, and filtering after extraction; 2) taking filter residues, adding the disodium hydrogen phosphate-citric acid buffer solution for regulating pH value to the range from 3.5 to 5.5, and after mixing, adding an enzyme for digesting plant cell walls, and filtering after enzymolysis; 3) taking filter residues, adding the disodium hydrogen phosphate-citric acid buffer solution for regulating pH value to the range from 7.0 to 8.0, and filtering after extraction; 4) taking filter residues, adding the disodium hydrogen phosphate-citric acid buffer solution for regulating pH value to the range from 8.1 to 8.5, and filtering after extraction; and 5) blending the filtrates of the steps 1)-4), recovering the solvent and then drying the fluid extract in vacuum, thereby obtaining dry extract containing hovenaia dulcis total alkaloids. Compared with independent use of the bionic method or the enzymic method, the method is high in extraction rate, mild in extraction condition, simple in operation, friendly to environment and suitable for industrial large-scale production.
Description
Technical field
The invention belongs to natural product extracting technique field, relate to the extractive technique field of total alkaloids, be specifically related to a kind of half bionical-method of Enzymatic Extraction honey raisin tree seven total alkaloidss.
Background technology
Cardiovascular and cerebrovascular disease are a kind of serious threat Human diseases, particularly the healthy common disease of more than 50 years old the elderly.Cardiovascular and cerebrovascular diseases has " sickness rate is high, disability rate is high, mortality ratio is high, recurrence rate is high, and complication is many " i.e. feature of " four is high by more than ".At present, the number of cardiovascular and cerebrovascular diseases is died from every year up to 1,500 ten thousand people in the whole world, occupies various causes of the death first place.Therefore, cardiovascular and cerebrovascular diseases is known as the No.1 killer that mankind's Death causes is the highest.
Honey raisin tree seven is the root and rhizome of bloodroot root of Japanese Hylomecon, and main alkaloid constituents is: protopine, cryptopine, coptisine, Berberine, sanguinarine(e), white chelerythrine, stylopine etc.These alkaloids are isoquinoline alkaloid substantially; Recent study report finds that this Alkaloid has multiple effect to cardiovascular systems; such as: from honey raisin tree seven, separating the protopine obtaining has provide protection to Myocardial Ischemia-reperfusion, has wide practical use clinically.
Summary of the invention
The problem that the present invention solves be to provide a kind of half bionical-method of Enzymatic Extraction honey raisin tree seven total alkaloidss, present method raw material sources are extensive, easy and simple to handle, extraction yield is high, is applicable to the Production requirement of large-scale industrialization.
The present invention is achieved through the following technical solutions:
The method of half bionical-Enzymatic Extraction honey raisin tree seven total alkaloidss, comprises the following steps:
1) get the honey raisin tree seven of pulverizing, by the solid-liquid ratio of 1g:8～10ml, add Sodium phosphate dibasic-citrate buffer solution, regulating pH value is 1.5～3.0, fully mix, and after water-bath is extracted, filtration;
2) get the filter residue after step 1) is filtered, add Sodium phosphate dibasic-citrate buffer solution, regulating pH value is 3.5～5.5, adds the enzyme of clearing up plant cell wall, water enzyme digestion at 50～60 DEG C after mixing; The consumption of the described enzyme of clearing up plant cell wall is 0.3～0.6% of honey raisin tree seven quality;
3) get step 2) filter residue after filtering, add Sodium phosphate dibasic-citrate buffer solution, regulating pH value is 7.0～8.0, fully mix, after water-bath is extracted, filtration;
4) get the filter residue after step 3) is filtered, add Sodium phosphate dibasic-citrate buffer solution, regulating pH value is 8.1～8.5, fully mixes, and after water-bath is extracted, filters;
5) by step 1)～4) filter the filtrate merging obtaining, reclaim after solvent, by the fluid extract vacuum-drying obtaining, obtain the dry extract that comprises honey raisin tree seven total alkaloidss.
The described enzyme of clearing up plant cell wall is cellulase, beta-glucanase or extracts prozyme.
Step 1), 3), 4) to extract temperature be 50～60 DEG C in described water-bath.
Step 1), 3), 4) described water-bath extraction time is 1～2h.
Step 2) the described water enzyme digestion time is 1～2h.
Vacuum-drying described in step 5) is to carry out at 60～80 DEG C.
Compared with prior art, the present invention has following useful technique effect:
That the present invention adopts is half bionical-method extract total alkaloids from honey raisin tree seven that enzyme process combines, half bionical-enzyme process is that semi-bionic extraction and biological enzyme are combined to a kind of novel method of extracting active ingredient of natural product, gather the advantage of half biomimetic method and Enzymatic Extraction technology, half biomimetic method meets traditional Chinese medical science compatibility at leaching process, the feature that clinical application and oral pharmaceutical absorb in gi tract transhipment, the present invention adopts the extraction solvent of Sodium phosphate dibasic-citric acid buffer system as half bionical-enzyme process, by different pH value gradients is set, reach aids drug and transport in vivo the object of absorption, in leaching process, add the applicable enzyme that can clear up plant cell wall, this enzyme is as a kind of biological catalyst, the effective constituent of herbal medicine cell walls is decomposed to destruction, reduce resistance to mass transfer, the optimal reactive temperature that regulates this enzyme is 50～60 DEG C, after enzymolysis, be more conducive to the extraction of total alkaloids, thereby raising extraction efficiency.
Half bionical-enzyme process is the further imitation biochemistry of half biomimetic method, the total alkaloids in the present invention adopts half bionical-Enzymatic Extraction honey raisin tree seven, and compared with independent use half biomimetic method, enzyme process, extraction yield increases.Extraction conditions gentleness of the present invention, simple to operate, environmental friendliness, be applicable to large-scale industrialization and produce, be with a wide range of applications.
Embodiment
Below in conjunction with specific embodiment and comparative example, the present invention is described in further detail, and the explanation of the invention is not limited.
Honey raisin tree seven medicinal materials described in embodiments of the invention and comparative example, are selected from papaveracease, and root of Japanese Hylomecon belongs to, and root of Japanese Hylomecon is dried root and rhizome, calls as root of Japanese Hylomecon root, sword bean pseudo-ginseng, water Chang pseudo-ginseng, great Ye LAOSHUQI, black muscle seven etc.
Embodiment 1
Half bionical-enzyme process (select and extract prozyme) extracts the method for total alkaloids in honey raisin tree seven, comprises the following steps:
1) get honey raisin tree seven medicinal materials after 10g pulverizes, Gl tract pH value, adds 100ml Sodium phosphate dibasic-citric acid solution, and adjust pH is 2.0, and fully concussion shakes up, and water-bath 2h at 60 DEG C of temperature filters.
2) in the filter residue obtaining to step 1), add Sodium phosphate dibasic-citric acid solution, adjust pH is 4.5, add extraction prozyme (to extract prozyme purchased from Ningxia jade of the He family Bioisystech Co., Ltd, comprise beta-glucanase, cellulase, zytase and polygalacturonase, wherein, the vigor of various enzymes is: beta glucan Mei≤3,000,000 u/g, Xian ties up Su Mei≤400,000 u/g, Mu Ju Tang Mei≤2,600,000 u/g, Guo Jiao Mei≤20,000 u/g) 0.05g, 50 DEG C of water enzyme digestion 2h, filter.
3) to step 2) continue to add Sodium phosphate dibasic-citric acid solution in the filter residue that obtains, adjust pH is 7.5, water-bath 2h at 60 DEG C filters.
4) in the filter residue obtaining to step 3), continue to add Sodium phosphate dibasic-citric acid solution, adjust pH is 8.3, and water-bath 2h at 60 DEG C filters.
5) combining step 1)～4) all filtrate, reclaim solvent and obtain total fluid extract, fluid extract, 80 DEG C of vacuum-dryings, is obtained to the dry extract 7.2564g that comprises honey raisin tree seven total alkaloidss.
Honey raisin tree seven Determination of Total Alkaloid:
1) precision takes white chelerythrine standard substance 1.0mg and is placed in the volumetric flask of 10ml, with (v/v) solution constant volume of hydrochloric acid-70% methyl alcohol (0.5:100), obtains reference liquid.
2) precision takes dry extract 6.7mg, is placed in the volumetric flask of 5ml, and use hydrochloric acid-70% methyl alcohol (0.5:100) (v/v) solution constant volume, to scale, obtains sample liquid.
3) measure respectively 0.5ml reference liquid and 0.5ml sample liquid, add respectively wherein 5ml citric acid-sodium citrate buffering test solution, 2ml tetrabromophenol sulfonphthalein test solution and 9.5ml methylene dichloride, get dichloromethane layer, with after anhydrous sodium sulfate dehydration, using methylene dichloride as reference, at 200～800nm scanning wavelength, the two all has maximum absorption at 287nm place.
4) precision takes dry extract 6.7mg, is placed in the volumetric flask of 5ml, with concentrated hydrochloric acid-70% methyl alcohol (0.5:100) (v/v) solution constant volume to scale.
5) precision measures 0.5ml sample liquid, add respectively 5ml citric acid-sodium citrate buffering test solution, 2ml tetrabromophenol sulfonphthalein test solution and 9.5ml methylene dichloride, get dichloromethane layer, with after anhydrous sodium sulfate dehydration, using methylene dichloride as reference, measure absorbancy at 287nm place, the content that calculates total alkaloids is 4.7842mg/g.
Embodiment 2
Half bionical-enzyme process (selecting cellulase) extracts the method for total alkaloids in honey raisin tree seven, comprises the following steps:
1) get honey raisin tree seven medicinal materials that 10g pulverizes, Gl tract pH value, adds 80ml Sodium phosphate dibasic-citric acid solution, and adjust pH is 2.0, and fully concussion shakes up, and water-bath 2h at 60 DEG C filters.
2) in the filter residue obtaining to step 1), continue to add Sodium phosphate dibasic-citric acid solution, adjust pH is 4.5, adds cellulase 0.05g, and 50 DEG C of water enzyme digestion 2h filter.
3) to step 2) continue to add Sodium phosphate dibasic-citric acid solution in the filter residue that obtains, adjust pH is 7.5, water-bath 2h at 60 DEG C filters.
4) in the filter residue obtaining to step 3), continue to add Sodium phosphate dibasic-citric acid solution, adjust pH is 8.3, and water-bath 2h at 60 DEG C filters.
5) combining step 1)～4) all filtrate, reclaim solvent and obtain total fluid extract, fluid extract, 80 DEG C of vacuum-dryings, is obtained to the dry extract 6.3274g that comprises honey raisin tree seven total alkaloidss.
Honey raisin tree seven Determination of Total Alkaloid:
1) precision takes dry extract 6.7mg, is placed in the volumetric flask of 5ml, and use hydrochloric acid-70% methyl alcohol (0.5:100) (v/v) solution constant volume, to scale, obtains sample liquid.
2) precision measures 0.5ml sample liquid, add respectively 5ml citric acid-sodium citrate buffering test solution, 2ml tetrabromophenol sulfonphthalein test solution and 9.5ml methylene dichloride, get dichloromethane layer, with after anhydrous sodium sulfate dehydration, using methylene dichloride as reference, measure absorbancy at 287nm place, the content that calculates total alkaloids is 3.9319mg/g.
Embodiment 3
Half bionical-enzyme process (selecting beta-glucanase) extracts the method for total alkaloids in honey raisin tree seven, comprises the following steps:
1) get honey raisin tree seven medicinal materials that 10g pulverizes, Gl tract pH value, adds 90ml Sodium phosphate dibasic-citric acid solution, and adjust pH is 2.5, and fully concussion shakes up, and water-bath 2h at 60 DEG C filters.
2) in the filter residue obtaining to step 1), continue to add Sodium phosphate dibasic-citric acid solution, adjust pH is 4.0, adds beta-glucanase 0.06g, and 50 DEG C of water enzyme digestion 2h filter.
3) to step 2) continue to add Sodium phosphate dibasic-citric acid solution in the filter residue that obtains, adjust pH is 7.0, water-bath 1.5h at 60 DEG C filters.
4) in the filter residue obtaining to step 3), continue to add Sodium phosphate dibasic-citric acid solution, adjust pH is 8.5, and water-bath 1.5h at 60 DEG C filters.
5) combining step 1)～4) all filtrate, reclaim solvent and obtain total fluid extract, fluid extract, 60 DEG C of vacuum-dryings, is obtained to the dry extract 6.1639g that comprises honey raisin tree seven total alkaloidss.
Embodiment 4
Half bionical-enzyme process (selecting beta-glucanase) extracts the method for total alkaloids in honey raisin tree seven, comprises the following steps:
1) get honey raisin tree seven medicinal materials that 10g pulverizes, Gl tract pH value, adds 80ml Sodium phosphate dibasic-citric acid solution, and adjust pH is 1.5, and fully concussion shakes up, and water-bath 1.5h at 60 DEG C filters.
2) in the filter residue obtaining to step 1), continue to add Sodium phosphate dibasic-citric acid solution, adjust pH is 3.5, adds beta-glucanase 0.03g, and 50 DEG C of water enzyme digestion 1h filter.
3) to step 2) continue to add Sodium phosphate dibasic-citric acid solution in the filter residue that obtains, adjust pH is 7.3, water-bath 1.5h at 60 DEG C filters.
4) in the filter residue obtaining to step 3), continue to add Sodium phosphate dibasic-citric acid solution, adjust pH is 8.1, and water-bath 1.5h at 60 DEG C filters.
5) combining step 1)～4) all filtrate, reclaim solvent and obtain total fluid extract, fluid extract, 70 DEG C of vacuum-dryings, is obtained to the dry extract 5.9715g that comprises honey raisin tree seven total alkaloidss.
Embodiment 5
Half bionical-enzyme process (select and extract prozyme) extracts the method for total alkaloids in honey raisin tree seven, comprises the following steps:
1) get honey raisin tree seven medicinal materials that 10g pulverizes, Gl tract pH value, adds 100ml Sodium phosphate dibasic-citric acid solution, and adjust pH is 3.0, and fully concussion shakes up, and water-bath 1h at 60 DEG C filters.
2) in the filter residue obtaining to step 1), continue to add Sodium phosphate dibasic-citric acid solution, adjust pH is 5.5, adds and extracts prozyme 0.04g, and 50 DEG C of water enzyme digestion 1.5h, filter.
3) to step 2) continue to add Sodium phosphate dibasic-citric acid solution in the filter residue that obtains, adjust pH is 8.0, water-bath 1h at 60 DEG C filters.
4) in the filter residue obtaining to step 3), continue to add Sodium phosphate dibasic-citric acid solution, adjust pH is 8.5, and water-bath 1h at 60 DEG C filters.
5) combining step 1)～4) all filtrate, reclaim solvent and obtain total fluid extract, fluid extract, 75 DEG C of vacuum-dryings, is obtained to the dry extract 6.7124g that comprises honey raisin tree seven total alkaloidss.
Be below the extraction comparative example that half biomimetic method, enzyme process use separately:
Comparative example 1:
Adopt enzyme process (selecting cellulase) to extract the total alkaloids in honey raisin tree seven, comprise the following steps:
1) get honey raisin tree seven medicinal materials of 10g through pulverizing, add 100ml Sodium phosphate dibasic-citric acid solution, adjust pH 4.5, then adds 0.05g cellulase, after 50 DEG C of water enzyme digestion 2h, filters;
2) filter residue after step 1) is filtered is with 70% ethanol water-bath 2 times at 60 DEG C, and each 1h, filters, merging filtrate, and recovery solvent obtains fluid extract, and fluid extract, 80 DEG C of vacuum-dryings, is obtained to dry extract 4.9835g.
Honey raisin tree seven Determination of Total Alkaloid:
1) precision takes white chelerythrine standard substance 1.0mg and is placed in the volumetric flask of 10ml, with (v/v) solution constant volume of hydrochloric acid-70% methyl alcohol (0.5:100), obtains reference liquid.
2) precision takes dry extract 6.7mg, is placed in the volumetric flask of 5ml, and use hydrochloric acid-70% methyl alcohol (0.5:100) (v/v) solution constant volume, to scale, obtains sample liquid.
3) measure respectively 0.5ml reference liquid and 0.5ml sample liquid, add respectively wherein 5ml citric acid-sodium citrate buffering test solution, 2ml tetrabromophenol sulfonphthalein test solution and 9.5ml methylene dichloride, get dichloromethane layer, with after anhydrous sodium sulfate dehydration, using methylene dichloride as reference, at 200～800nm scanning wavelength, the two all has maximum absorption at 287nm place.
4) precision measures 0.5ml sample liquid, add respectively 5ml citric acid-sodium citrate buffering test solution, 2ml tetrabromophenol sulfonphthalein test solution and 9.5ml methylene dichloride, get dichloromethane layer, with after anhydrous sodium sulfate dehydration, using methylene dichloride as reference, measure absorbancy at 287nm place, the content that calculates total alkaloids is 3.4130mg/g.
Comparative example 2
Adopt half biomimetic method to extract the total alkaloids in honey raisin tree seven, comprise the following steps:
1) get honey raisin tree seven medicinal materials of 10g through pulverizing, add 100ml Sodium phosphate dibasic-citric acid solution, adjust pH is 2.0, and fully concussion shakes up, and water-bath 2h at 60 DEG C filters.
2) filter in the filter residue obtaining and add Sodium phosphate dibasic-citric acid solution to step 1), adjust pH is 7.5, and water-bath 2h at 60 DEG C filters.
3) to step 2) filter in the filter residue obtaining and continue to add Sodium phosphate dibasic-citric acid solution, adjust pH is 8.3, water-bath 2h at 60 DEG C filters.
4) combining step 1), 2), 3) filtrate, reclaim solvent total fluid extract, fluid extract, 80 DEG C of vacuum-dryings, is obtained to the dry extract 5.5821g that comprises honey raisin tree seven total alkaloidss.
Honey raisin tree seven Determination of Total Alkaloid:
1) take dry extract 6.7mg obtained above, be placed in the volumetric flask of 5ml, use hydrochloric acid-70% methyl alcohol (0.5:100) (v/v) solution constant volume, to scale, is made into sample liquid.
2) precision measures 0.5ml sample liquid, add 5ml citric acid-sodium citrate buffering test solution, 2ml tetrabromophenol sulfonphthalein test solution and 9.5ml methylene dichloride, mix rear leaving standstill, get dichloromethane layer, with after anhydrous sodium sulfate dehydration, using methylene dichloride as reference, measuring absorbancy at 287nm place, calculate the content of total alkaloids, is 3.700mg/g.
Comparative example 3
Adopt enzyme process (select and extract prozyme) to extract the total alkaloids in honey raisin tree seven, comprise the following steps:
1) get honey raisin tree seven medicinal materials of 10g through pulverizing, add 100ml Sodium phosphate dibasic-citric acid solution adjust pH 4.5, then add 0.05g to extract prozyme (purchased from Ningxia jade of the He family Bioisystech Co., Ltd, comprise beta-glucanase, cellulase, zytase and polygalacturonase), after 50 DEG C of water enzyme digestion 2h, filter.
2) filter residue after step 1) is filtered is with 70% ethanol water-bath 2 times at 60 DEG C, and each 1h, filters, merging filtrate, and recovery solvent obtains fluid extract, and fluid extract, 80 DEG C of vacuum-dryings, is obtained to the dry extract 4.0919g that comprises honey raisin tree seven total alkaloidss.
Honey raisin tree seven Determination of Total Alkaloid:
1) precision takes dry extract 6.7mg, is placed in the volumetric flask of 5ml, and use hydrochloric acid-70% methyl alcohol (0.5:100) (v/v) solution constant volume, to scale, obtains sample liquid.
2) precision measures 0.5ml sample liquid, add 5ml citric acid-sodium citrate buffering test solution, 2ml tetrabromophenol sulfonphthalein test solution and 9.5ml methylene dichloride, get dichloromethane layer, with after anhydrous sodium sulfate dehydration, using methylene dichloride as reference, measure absorbancy at 287nm place, the content that calculates total alkaloids is 4.1451mg/g.
Comparative example 4
Adopt enzyme process (selecting beta-glucanase) to extract the total alkaloids in honey raisin tree seven, comprise the following steps:
1) get 10g honey raisin tree seven medicinal materials after pulverizing, add 100ml Sodium phosphate dibasic-citric acid solution, adjust pH 4.5, then adds 0.05g beta-glucanase, after 50 DEG C of water enzyme digestion 2h, filters.
2) by the filter residue after filtering at 60 DEG C, extract 2 times with 70% ethanol water-bath, each 1h, filters, merging filtrate, recovery solvent obtains fluid extract, and fluid extract, 80 DEG C of vacuum-dryings, is obtained to the dry extract 3.7264g that comprises honey raisin tree seven total alkaloidss.
Honey raisin tree seven Determination of Total Alkaloid:
1) precision takes dry extract 6.7mg, is placed in the volumetric flask of 5ml, and use hydrochloric acid-70% methyl alcohol (0.5:100) (v/v) solution constant volume, to scale, obtains sample liquid.
2) precision measures 0.5ml sample liquid, add 5ml citric acid-sodium citrate buffering test solution, 2ml tetrabromophenol sulfonphthalein test solution and 9.5ml methylene dichloride, get dichloromethane layer, with after anhydrous sodium sulfate dehydration, using methylene dichloride as reference, measure absorbancy at 287nm place, the content that calculates total alkaloids is 2.4333mg/g.
Can find out from embodiment and comparative example: half bionical-extracting combined-enzyme method has improved 29.30% than the total alkaloid content of half biomimetic method extraction; Half bionical-extracting combined-enzyme method than the total alkaloid content that extracts combined-enzyme method and extract has improved 15.42%; The total alkaloid content that half bionical-Mierocrystalline cellulose enzyme process extracts than half biomimetic method has improved 6.27%; The total alkaloid content that half bionical-Mierocrystalline cellulose enzyme process extracts than Mierocrystalline cellulose enzyme process has improved 15.2%; Half bionical-to extract combined-enzyme method higher by 21.68% than the total alkaloid content of half bionical-Mierocrystalline cellulose enzyme process extraction.Compared with biomimetic method, enzyme process, the extraction yield of half biomimetic enzyme method improves, and this has manifested the superiority of this new extracting method of half biomimetic enzyme method to a certain extent.
Extract and obtained the dry extract that comprises honey raisin tree seven total alkaloidss by the inventive method, adopt acid-dye colorimetry, using white chelerythrine as standard substance, the total alkaloids extracting in the dry extract obtaining is carried out to assay, honey raisin tree seven total alkaloidss comprise protopine, cryptopine, coptisine, Berberine, sanguinarine(e), white chelerythrine, chelilutine, Stylophorine, stylopine etc., the vital role that honey raisin tree seven total alkaloidss of reporting in view of Recent study play in treatment cardiovascular system diseases, the extraction efficiency that improves honey raisin tree seven total alkaloidss is significant.
Claims (5)
1. half is bionical-methods of Enzymatic Extraction honey raisin tree seven total alkaloidss, it is characterized in that, and comprise the following steps:
1) get the honey raisin tree seven of pulverizing, by the solid-liquid ratio of 1g:8～10ml, add Sodium phosphate dibasic-citrate buffer solution, regulating pH value is 1.5～3.0, fully mix, and after water-bath is extracted, filtration;
2) get the filter residue after step 1) is filtered, add Sodium phosphate dibasic-citrate buffer solution, regulating pH value is 3.5～5.5, adds the enzyme of clearing up plant cell wall, water enzyme digestion at 50～60 DEG C after mixing; The consumption of the described enzyme of clearing up plant cell wall is 0.3～0.6% of honey raisin tree seven quality;
Wherein, the described enzyme of clearing up plant cell wall is for extracting prozyme, extract prozyme and comprise beta-glucanase, cellulase, zytase and polygalacturonase, wherein, the vigor of various enzymes is: beta glucan Mei≤3,000,000 u/g, Xian ties up Su Mei≤400,000 u/g, Mu Ju Tang Mei≤2,600,000 u/g, Guo Jiao Mei≤20,000 u/g;
3) get step 2) filter residue after filtering, add Sodium phosphate dibasic-citrate buffer solution, regulating pH value is 7.0～8.0, fully mix, after water-bath is extracted, filtration;
4) get the filter residue after step 3) is filtered, add Sodium phosphate dibasic-citrate buffer solution, regulating pH value is 8.1～8.5, fully mixes, and after water-bath is extracted, filters;
5) by step 1)～4) filter the filtrate merging obtaining, reclaim after solvent, by the fluid extract vacuum-drying obtaining, obtain the dry extract that comprises honey raisin tree seven total alkaloidss.
According to claim 1 a kind of half bionical-method of Enzymatic Extraction honey raisin tree seven total alkaloidss, it is characterized in that: step 1), 3), 4) to extract temperature be 50～60 DEG C in described water-bath.
According to claim 1 a kind of half bionical-method of Enzymatic Extraction honey raisin tree seven total alkaloidss, it is characterized in that: step 1), 3), 4) described water-bath extraction time is 1～2h.
According to claim 1 a kind of half bionical-method of Enzymatic Extraction honey raisin tree seven total alkaloidss, it is characterized in that: step 2) the described water enzyme digestion time is 1～2h.
According to claim 1 a kind of half bionical-method of Enzymatic Extraction honey raisin tree seven total alkaloidss, it is characterized in that: the vacuum-drying described in step 5) is to carry out at 60～80 DEG C.
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