CN103202884A - 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 the natural product extracting technique field, relate to the extractive technique field of total alkaloids, be specifically related to a kind of method of half bionical-Enzymatic Extraction Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss.
Background technology
Cardiovascular and cerebrovascular disease is a kind of serious threat human diseases, particularly the commonly encountered diseases of middle-aged and elderly people health more than 50 years old.Cardiovascular and cerebrovascular disease has " sickness rate height, disability rate height, mortality rate height, relapse rate height, complication is many " i.e. characteristics of " four is high by more than ".At present, every year is died from the number of cardiovascular and cerebrovascular disease up to 1,500 ten thousand people in the whole world, occupies various causes of the death first place.Therefore, cardiovascular and cerebrovascular disease is become the highest No.1 killer of the human dead cause of disease.
Semen Hoveniae (Fructus Hoveniae) seven is the root and rhizome of bloodroot Radix Hylomeci japonicae, and the main alkaloid constituents is: protopine, cryptocavine, coptisine, berberine, Sanguinarine, chelerythrine, stylopine etc.These alkaloids are isoquinoline alkaloid substantially; the Recent study report finds that this Alkaloid has multiple effect to cardiovascular system; such as: separating the protopine that obtains from Semen Hoveniae (Fructus Hoveniae) seven has protective effect to the ischemia-reperfusion cardiac muscle, has wide practical use clinically.
Summary of the invention
The problem that the present invention solves is to provide a kind of method of half bionical-Enzymatic Extraction Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss, and this method raw material sources are extensive, and are easy and simple to handle, and the extraction ratio height is fit to industrial and large-scale production demand.
The present invention is achieved through the following technical solutions:
The method of half bionical-Enzymatic Extraction Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss may further comprise the steps:
1) get the Semen Hoveniae (Fructus Hoveniae) seven of pulverizing, press the solid-liquid ratio of 1g:8～10ml, add sodium hydrogen phosphate-citrate buffer solution, regulating pH value is 1.5～3.0, and fully mixing after water-bath is extracted, filters;
2) get filtering residue after step 1) is filtered, add sodium hydrogen phosphate-citrate buffer solution, regulating pH value is 3.5～5.5, adds the enzyme of clearing up plant cell wall behind the mixing, at 50～60 ℃ of following water enzyme digestions; The described consumption of clearing up the enzyme of plant cell wall is 0.3～0.6% of Semen Hoveniae (Fructus Hoveniae) seven quality;
3) get step 2) filtering residue after filtering, add sodium hydrogen phosphate-citrate buffer solution, regulating pH value is 7.0～8.0, fully mixing after water-bath is extracted, filters;
4) get filtering residue after step 3) is filtered, add sodium hydrogen phosphate-citrate buffer solution, regulating pH value is 8.1～8.5, and fully mixing after water-bath is extracted, filters;
5) with step 1)～4) filter the filtrate merging that obtains, behind the recovery solvent, with the fluid extract vacuum drying that obtains, obtain comprising the dry extract of Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss.
The described enzyme of clearing up plant cell wall is cellulase, 1,4 beta-glucanase or extracts compound enzyme.
Step 1), 3), 4) described water-bath extraction temperature is 50～60 ℃.
Step 1), 3), 4) described water-bath extraction time is 1～2h.
Step 2) described water enzyme digestion time is 1～2h.
The described vacuum drying of step 5) is to carry out under 60～80 ℃.
Compared with prior art, the present invention has following beneficial technical effects:
Method extract total alkaloids from Semen Hoveniae (Fructus Hoveniae) seven that the present invention adopts half bionical-enzyme process to combine, half bionical-enzyme process is that semi-bionic extraction and enzyme are combined a kind of new method of extracting active ingredient of natural product, the advantage of having gathered semi-biomimetic method and Enzymatic Extraction technology, semi-biomimetic method meets traditional Chinese medical science compatibility at leaching process, the characteristics that clinical application and oral drugs absorb in the gastrointestinal tract transhipment, the present invention adopts sodium hydrogen phosphate-citric acid buffer system as the extraction solvent of half bionical-enzyme process, by different pH value gradients is set, reach aids drug and transport the purpose of absorption in vivo, in leaching process, add the enzyme that can clear up plant cell wall that is fit to, this enzyme is as a kind of biocatalyzer, effective ingredient to the Chinese herbal medicine cell wall decomposes destruction, reduce resistance to mass tranfer, the optimal reactive temperature of regulating this enzyme is 50～60 ℃, be more conducive to the extraction of total alkaloids behind the enzymolysis, thereby improve extraction efficiency.
Half bionical-enzyme process is the further imitation biochemistry of semi-biomimetic method, and the present invention adopts the total alkaloids in half bionical-Enzymatic Extraction Semen Hoveniae (Fructus Hoveniae) seven, compares with independent use semi-biomimetic method, enzyme process, and extraction ratio increases.Extraction conditions gentleness of the present invention, simple to operate, environmental friendliness, be fit to large-scale industrialization production, be with a wide range of applications.
The specific embodiment
The present invention is described in further detail below in conjunction with specific embodiment and Comparative Examples, and the explanation of the invention is not limited.
Semen Hoveniae (Fructus Hoveniae) seven medical materials described in embodiments of the invention and the Comparative Examples are selected from Papaveraceae, and Radix Hylomeci japonicae belongs to, and the dry root and rhizome of Radix Hylomeci japonicae is called and is Radix Hylomeci japonicae root, Semen Canavaliae Radix Notoginseng, water Chang Radix Notoginseng, Da Ye Hylomecon japonica, black muscle seven etc.
Embodiment 1
Half bionical-enzyme process (select for use and extract compound enzyme) extracts the method for total alkaloids in the Semen Hoveniae (Fructus Hoveniae) seven, may further comprise the steps:
1) Semen Hoveniae (Fructus Hoveniae) seven medical materials after getting 10g and pulverizing, the Gl tract pH value adds 100ml sodium hydrogen phosphate-citric acid solution, and adjust pH is 2.0, and fully concussion shakes up, and water-bath 2h under 60 ℃ of temperature filters.
2) add sodium hydrogen phosphate-citric acid solution in the filtering residue that obtains to step 1), adjust pH is 4.5, adding the extraction compound enzyme (extracts compound enzyme available from Ningxia jade of the He family Bioisystech Co., Ltd, comprises 1,4 beta-glucanase, cellulase, xylanase and pectase, wherein, the vigor of various enzymes is: beta glucan Mei ≧ 3,000,000 u/g, Xian dimension 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 ℃ of water enzyme digestion 2h filter.
3) to step 2) continue to add sodium hydrogen phosphate-citric acid solution in the filtering residue that obtains, adjust pH is 7.5, at 60 ℃ of following water-bath 2h, filters.
4) continue in the filtering residue that obtains to step 3) to add sodium hydrogen phosphate-citric acid solution, adjust pH is 8.3, at 60 ℃ of following water-bath 2h, filters.
5) combining step 1)～4) all filtrate, reclaim solvent and get total fluid extract, fluid extract at 80 ℃ of vacuum dryings, is obtained comprising the dry extract 7.2564g of Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss.
Semen Hoveniae (Fructus Hoveniae) seven Determination of Total Alkaloid:
1) precision takes by weighing the volumetric flask that chelerythrine standard substance 1.0mg places 10ml, uses (v/v) solution standardize solution of hydrochloric acid-70% methanol (0.5:100), obtains titer.
2) precision takes by weighing dry extract 6.7mg, places the volumetric flask of 5ml, and usefulness hydrochloric acid-70% methanol (0.5:100) (v/v) solution standardize solution obtains sample liquid to scale.
3) measure 0.5ml titer and 0.5ml sample liquid respectively, to wherein adding 5ml citric acid-sodium citrate buffering test solution respectively, 2ml bromophenol blue test solution and 9.5ml dichloromethane, get dichloromethane layer, behind anhydrous sodium sulfate dehydration, as reference, at 200～800nm scanning wavelength, the two all has absorption maximum at the 287nm place with dichloromethane.
4) precision takes by weighing dry extract 6.7mg, places the volumetric flask of 5ml, with concentrated hydrochloric acid-70% methanol (0.5:100) (v/v) the solution standardize solution to scale.
5) precision is measured the 0.5ml sample liquid, add 5ml citric acid-sodium citrate buffering test solution respectively, 2ml bromophenol blue test solution and 9.5ml dichloromethane, get dichloromethane layer, behind anhydrous sodium sulfate dehydration, as reference, measure absorbance with dichloromethane at the 287nm place, calculate total alkaloid contents, be 4.7842mg/g.
Embodiment 2
Half bionical-enzyme process (selecting cellulase for use) extracts the method for total alkaloids in the Semen Hoveniae (Fructus Hoveniae) seven, may further comprise the steps:
1) get Semen Hoveniae (Fructus Hoveniae) seven medical materials that 10g pulverizes, the Gl tract pH value adds 80ml sodium hydrogen phosphate-citric acid solution, and adjust pH is 2.0, and fully concussion shakes up, and at 60 ℃ of following water-bath 2h, filters.
2) continue in the filtering residue that obtains to step 1) to add sodium hydrogen phosphate-citric acid solution, adjust pH is 4.5, adds cellulase 0.05g, and 50 ℃ of water enzyme digestion 2h filter.
3) to step 2) continue to add sodium hydrogen phosphate-citric acid solution in the filtering residue that obtains, adjust pH is 7.5, at 60 ℃ of following water-bath 2h, filters.
4) continue in the filtering residue that obtains to step 3) to add sodium hydrogen phosphate-citric acid solution, adjust pH is 8.3, at 60 ℃ of following water-bath 2h, filters.
5) combining step 1)～4) all filtrate, reclaim solvent and get total fluid extract, fluid extract at 80 ℃ of vacuum dryings, is obtained comprising the dry extract 6.3274g of Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss.
Semen Hoveniae (Fructus Hoveniae) seven Determination of Total Alkaloid:
1) precision takes by weighing dry extract 6.7mg, places the volumetric flask of 5ml, and usefulness hydrochloric acid-70% methanol (0.5:100) (v/v) solution standardize solution obtains sample liquid to scale.
2) precision is measured the 0.5ml sample liquid, add 5ml citric acid-sodium citrate buffering test solution respectively, 2ml bromophenol blue test solution and 9.5ml dichloromethane, get dichloromethane layer, behind anhydrous sodium sulfate dehydration, as reference, measure absorbance with dichloromethane at the 287nm place, calculate total alkaloid contents, be 3.9319mg/g.
Embodiment 3
Half bionical-enzyme process (selecting 1,4 beta-glucanase for use) extracts the method for total alkaloids in the Semen Hoveniae (Fructus Hoveniae) seven, may further comprise the steps:
1) get Semen Hoveniae (Fructus Hoveniae) seven medical materials that 10g pulverizes, the Gl tract pH value adds 90ml sodium hydrogen phosphate-citric acid solution, and adjust pH is 2.5, and fully concussion shakes up, and at 60 ℃ of following water-bath 2h, filters.
2) continue in the filtering residue that obtains to step 1) to add sodium hydrogen phosphate-citric acid solution, adjust pH is 4.0, adds 1,4 beta-glucanase 0.06g, and 50 ℃ of water enzyme digestion 2h filter.
3) to step 2) continue to add sodium hydrogen phosphate-citric acid solution in the filtering residue that obtains, adjust pH is 7.0, at 60 ℃ of following water-bath 1.5h, filters.
4) continue in the filtering residue that obtains to step 3) to add sodium hydrogen phosphate-citric acid solution, adjust pH is 8.5, at 60 ℃ of following water-bath 1.5h, filters.
5) combining step 1)～4) all filtrate, reclaim solvent and get total fluid extract, fluid extract at 60 ℃ of vacuum dryings, is obtained comprising the dry extract 6.1639g of Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss.
Embodiment 4
Half bionical-enzyme process (selecting 1,4 beta-glucanase for use) extracts the method for total alkaloids in the Semen Hoveniae (Fructus Hoveniae) seven, may further comprise the steps:
1) get Semen Hoveniae (Fructus Hoveniae) seven medical materials that 10g pulverizes, the Gl tract pH value adds 80ml sodium hydrogen phosphate-citric acid solution, and adjust pH is 1.5, and fully concussion shakes up, and at 60 ℃ of following water-bath 1.5h, filters.
2) continue in the filtering residue that obtains to step 1) to add sodium hydrogen phosphate-citric acid solution, adjust pH is 3.5, adds 1,4 beta-glucanase 0.03g, and 50 ℃ of water enzyme digestion 1h filter.
3) to step 2) continue to add sodium hydrogen phosphate-citric acid solution in the filtering residue that obtains, adjust pH is 7.3, at 60 ℃ of following water-bath 1.5h, filters.
4) continue in the filtering residue that obtains to step 3) to add sodium hydrogen phosphate-citric acid solution, adjust pH is 8.1, at 60 ℃ of following water-bath 1.5h, filters.
5) combining step 1)～4) all filtrate, reclaim solvent and get total fluid extract, fluid extract at 70 ℃ of vacuum dryings, is obtained comprising the dry extract 5.9715g of Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss.
Embodiment 5
Half bionical-enzyme process (select for use and extract compound enzyme) extracts the method for total alkaloids in the Semen Hoveniae (Fructus Hoveniae) seven, may further comprise the steps:
1) get Semen Hoveniae (Fructus Hoveniae) seven medical materials that 10g pulverizes, the Gl tract pH value adds 100ml sodium hydrogen phosphate-citric acid solution, and adjust pH is 3.0, and fully concussion shakes up, and at 60 ℃ of following water-bath 1h, filters.
2) continue in the filtering residue that obtains to step 1) to add sodium hydrogen phosphate-citric acid solution, adjust pH is 5.5, adds and extracts compound enzyme 0.04g, and 50 ℃ of water enzyme digestion 1.5h filter.
3) to step 2) continue to add sodium hydrogen phosphate-citric acid solution in the filtering residue that obtains, adjust pH is 8.0, at 60 ℃ of following water-bath 1h, filters.
4) continue in the filtering residue that obtains to step 3) to add sodium hydrogen phosphate-citric acid solution, adjust pH is 8.5, at 60 ℃ of following water-bath 1h, filters.
5) combining step 1)～4) all filtrate, reclaim solvent and get total fluid extract, fluid extract at 75 ℃ of vacuum dryings, is obtained comprising the dry extract 6.7124g of Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss.
Below be the extraction Comparative Examples that semi-biomimetic method, enzyme process use separately:
Comparative Examples 1:
Adopt the total alkaloids in enzyme process (selecting cellulase for use) the extraction Semen Hoveniae (Fructus Hoveniae) seven, may further comprise the steps:
1) get Semen Hoveniae (Fructus Hoveniae) seven medical materials of 10g through pulverizing, add 100ml sodium hydrogen phosphate-citric acid solution, adjust pH 4.5 adds the 0.05g cellulase then, behind 50 ℃ of water enzyme digestion 2h, filters;
2) filtering residue after step 1) is filtered 60 ℃ down with 70% ethanol water-bath 2 times, each 1h filters, merging filtrate, the recovery solvent gets fluid extract, and fluid extract at 80 ℃ of vacuum dryings, is obtained dry extract 4.9835g.
Semen Hoveniae (Fructus Hoveniae) seven Determination of Total Alkaloid:
1) precision takes by weighing the volumetric flask that chelerythrine standard substance 1.0mg places 10ml, uses (v/v) solution standardize solution of hydrochloric acid-70% methanol (0.5:100), obtains titer.
2) precision takes by weighing dry extract 6.7mg, places the volumetric flask of 5ml, and usefulness hydrochloric acid-70% methanol (0.5:100) (v/v) solution standardize solution obtains sample liquid to scale.
3) measure 0.5ml titer and 0.5ml sample liquid respectively, to wherein adding 5ml citric acid-sodium citrate buffering test solution respectively, 2ml bromophenol blue test solution and 9.5ml dichloromethane, get dichloromethane layer, behind anhydrous sodium sulfate dehydration, as reference, at 200～800nm scanning wavelength, the two all has absorption maximum at the 287nm place with dichloromethane.
4) precision is measured the 0.5ml sample liquid, add 5ml citric acid-sodium citrate buffering test solution respectively, 2ml bromophenol blue test solution and 9.5ml dichloromethane, get dichloromethane layer, behind anhydrous sodium sulfate dehydration, as reference, measure absorbance with dichloromethane at the 287nm place, calculate total alkaloid contents, be 3.4130mg/g.
Comparative Examples 2
Adopt the total alkaloids in the semi-biomimetic method extraction Semen Hoveniae (Fructus Hoveniae) seven, may further comprise the steps:
1) get Semen Hoveniae (Fructus Hoveniae) seven medical materials of 10g through pulverizing, add 100ml sodium hydrogen phosphate-citric acid solution, adjust pH is 2.0, and fully concussion shakes up, and at 60 ℃ of following water-bath 2h, filters.
2) filter in the filtering residue obtain to step 1) and add sodium hydrogen phosphate-citric acid solution, adjust pH is 7.5, at 60 ℃ of following water-bath 2h, filters.
3) to step 2) filter and continue in the filtering residue that obtains to add sodium hydrogen phosphate-citric acid solution, adjust pH is 8.3, at 60 ℃ of following water-bath 2h, filters.
4) combining step 1), 2), 3) filtrate, reclaim solvent total fluid extract, fluid extract at 80 ℃ of vacuum dryings, is obtained comprising the dry extract 5.5821g of Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss.
Semen Hoveniae (Fructus Hoveniae) seven Determination of Total Alkaloid:
1) take by weighing the above-mentioned dry extract 6.7mg that obtains, place the volumetric flask of 5ml, usefulness hydrochloric acid-70% methanol (0.5:100) (v/v) solution standardize solution is made into sample liquid to scale.
2) precision is measured the 0.5ml sample liquid, add 5ml citric acid-sodium citrate buffering test solution, 2ml bromophenol blue test solution and 9.5ml dichloromethane, leave standstill behind the mixing, get dichloromethane layer, behind anhydrous sodium sulfate dehydration, with dichloromethane as reference, measure absorbance at the 287nm place, calculate total alkaloid contents, be 3.700mg/g.
Comparative Examples 3
Adopt enzyme process (select for use and extract compound enzyme) to extract the total alkaloids in the Semen Hoveniae (Fructus Hoveniae) seven, may further comprise the steps:
1) gets Semen Hoveniae (Fructus Hoveniae) seven medical materials of 10g through pulverizing, add 100ml sodium hydrogen phosphate-citric acid solution adjust pH 4.5, add 0.05g then and extract compound enzyme (available from Ningxia jade of the He family Bioisystech Co., Ltd, comprise 1,4 beta-glucanase, cellulase, xylanase and pectase), behind 50 ℃ of water enzyme digestion 2h, filter.
2) filtering residue after step 1) is filtered 60 ℃ down with 70% ethanol water-bath 2 times, each 1h filters, merging filtrate, the recovery solvent gets fluid extract, and fluid extract at 80 ℃ of vacuum dryings, is obtained comprising the dry extract 4.0919g of Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss.
Semen Hoveniae (Fructus Hoveniae) seven Determination of Total Alkaloid:
1) precision takes by weighing dry extract 6.7mg, places the volumetric flask of 5ml, and usefulness hydrochloric acid-70% methanol (0.5:100) (v/v) solution standardize solution obtains sample liquid to scale.
2) precision is measured the 0.5ml sample liquid, add 5ml citric acid-sodium citrate buffering test solution, 2ml bromophenol blue test solution and 9.5ml dichloromethane, get dichloromethane layer, behind anhydrous sodium sulfate dehydration, as reference, measure absorbance with dichloromethane at the 287nm place, calculate total alkaloid contents, be 4.1451mg/g.
Comparative Examples 4
Adopt the total alkaloids in enzyme process (selecting 1,4 beta-glucanase for use) the extraction Semen Hoveniae (Fructus Hoveniae) seven, may further comprise the steps:
1) get 10g Semen Hoveniae (Fructus Hoveniae) seven medical materials after the pulverizing, add 100ml sodium hydrogen phosphate-citric acid solution, adjust pH 4.5 adds the 0.05g 1,4 beta-glucanase then, filters behind 50 ℃ of water enzyme digestion 2h.
2) filtering residue after will filtering extracts 2 times with 70% ethanol water-bath under 60 ℃, and each 1h filters, and merging filtrate reclaims solvent and gets fluid extract, and fluid extract at 80 ℃ of vacuum dryings, is obtained comprising the dry extract 3.7264g of Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss.
Semen Hoveniae (Fructus Hoveniae) seven Determination of Total Alkaloid:
1) precision takes by weighing dry extract 6.7mg, places the volumetric flask of 5ml, and usefulness hydrochloric acid-70% methanol (0.5:100) (v/v) solution standardize solution obtains sample liquid to scale.
2) precision is measured the 0.5ml sample liquid, add 5ml citric acid-sodium citrate buffering test solution, 2ml bromophenol blue test solution and 9.5ml dichloromethane, get dichloromethane layer, behind anhydrous sodium sulfate dehydration, as reference, measure absorbance with dichloromethane at the 287nm place, calculate total alkaloid contents, be 2.4333mg/g.
From embodiment and Comparative Examples as can be seen: half bionical-extracting combined-enzyme method has improved 29.30% than the total alkaloid content that semi-biomimetic method extracts; Half bionical-extraction combined-enzyme method has improved 15.42% than the total alkaloid content that extracts the combined-enzyme method extraction; Half bionical-cellulose enzyme process has improved 6.27% than the total alkaloid content that semi-biomimetic method extracts; Half bionical-cellulose enzyme process has improved 15.2% than the total alkaloid content that cellulose enzyme process extracts; Half bionical-extraction combined-enzyme method is higher by 21.68% than the total alkaloid content that half bionical-cellulose enzyme process extracts.Compare with biomimetic method, enzyme process, the extraction ratio of half bionical enzyme process improves, and this has manifested the superiority of this new extracting method of half bionical enzyme process to a certain extent.
Extract the dry extract that has obtained comprising Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss by the inventive method, adopt acid-dye colorimetry, with chelerythrine as standard substance, total alkaloids in the dry extract that extraction is obtained carries out assay, Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss comprise protopine, cryptocavine, coptisine, berberine, Sanguinarine, chelerythrine, chelilutine, Chelidonine, stylopine etc., the important function that Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss of reporting in view of Recent study play in the treatment cardiovascular system diseases, the extraction efficiency that improves Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss is significant.
Claims (6)
1. one and half is bionical-method of Enzymatic Extraction Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss, it is characterized in that, and may further comprise the steps:
1) get the Semen Hoveniae (Fructus Hoveniae) seven of pulverizing, press the solid-liquid ratio of 1g:8～10ml, add sodium hydrogen phosphate-citrate buffer solution, regulating pH value is 1.5～3.0, and fully mixing after water-bath is extracted, filters;
2) get filtering residue after step 1) is filtered, add sodium hydrogen phosphate-citrate buffer solution, regulating pH value is 3.5～5.5, adds the enzyme of clearing up plant cell wall behind the mixing, at 50～60 ℃ of following water enzyme digestions; The described consumption of clearing up the enzyme of plant cell wall is 0.3～0.6% of Semen Hoveniae (Fructus Hoveniae) seven quality;
3) get step 2) filtering residue after filtering, add sodium hydrogen phosphate-citrate buffer solution, regulating pH value is 7.0～8.0, fully mixing after water-bath is extracted, filters;
4) get filtering residue after step 3) is filtered, add sodium hydrogen phosphate-citrate buffer solution, regulating pH value is 8.1～8.5, and fully mixing after water-bath is extracted, filters;
5) with step 1)～4) filter the filtrate merging that obtains, behind the recovery solvent, with the fluid extract vacuum drying that obtains, obtain comprising the dry extract of Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss.
2. the method for a kind of half bionical-Enzymatic Extraction Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss according to claim 1, it is characterized in that: the described enzyme of clearing up plant cell wall is cellulase, 1,4 beta-glucanase or extracts compound enzyme.
3. the method for a kind of half bionical-Enzymatic Extraction Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss according to claim 1 is characterized in that: step 1), 3), 4) to extract temperature be 50～60 ℃ in described water-bath.
4. the method for a kind of half bionical-Enzymatic Extraction Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss according to claim 1 is characterized in that: step 1), 3), 4) described water-bath extraction time is 1～2h.
5. the method for a kind of half bionical-Enzymatic Extraction Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss according to claim 1 is characterized in that: step 2) the described water enzyme digestion time is 1～2h.
6. the method for a kind of half bionical-Enzymatic Extraction Semen Hoveniae (Fructus Hoveniae) seven total alkaloidss according to claim 1, it is characterized in that: the described vacuum drying of step 5) is to carry out under 60～80 ℃.
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| CN104370814A (en) * | 2014-06-20 | 2015-02-25 | 西安交通大学 | Method for extracting glaucine from pachysandra terminalis Sieb.et Zucc. |
| CN105820172A (en) * | 2016-03-23 | 2016-08-03 | 安徽农业大学 | Preparation method of alkaloid hydrochloride with anesthetic effect |
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| CN110051596A (en) * | 2019-04-24 | 2019-07-26 | 武汉科技大学 | A kind of dendrobium nobile toothpaste of antibacterial anti-inflammatory |
| CN115636883A (en) * | 2022-09-05 | 2023-01-24 | 溯玄(上海)生物技术有限公司 | Plant purification component, preparation method, skin repair activity and application of plant purification component in preparation of skin repair cosmetics |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104370814A (en) * | 2014-06-20 | 2015-02-25 | 西安交通大学 | Method for extracting glaucine from pachysandra terminalis Sieb.et Zucc. |
| CN104127509A (en) * | 2014-07-22 | 2014-11-05 | 西安交通大学 | Method for extracting total anthraquinone from Rhizoma Polygoni Cuspidati through aqueous enzymatic process-semi-bionic process |
| CN104127509B (en) * | 2014-07-22 | 2016-04-27 | 西安交通大学 | A kind of aqueous enzymatic method-semi-biomimetic method extracts the method for general anthraquinone in Rhizoma Polygoni Cuspidati |
| CN105820172A (en) * | 2016-03-23 | 2016-08-03 | 安徽农业大学 | Preparation method of alkaloid hydrochloride with anesthetic effect |
| CN105832789A (en) * | 2016-05-16 | 2016-08-10 | 威海松龄诺可佳中药饮片股份有限公司 | Method for preparing saponin enriched product through high-pressure microwave assisted semi-bionic extraction |
| WO2019080571A1 (en) * | 2017-10-24 | 2019-05-02 | 江苏金太阳纺织科技股份有限公司 | Method for extracting toosendanin |
| CN110051596A (en) * | 2019-04-24 | 2019-07-26 | 武汉科技大学 | A kind of dendrobium nobile toothpaste of antibacterial anti-inflammatory |
| CN110051596B (en) * | 2019-04-24 | 2022-05-17 | 武汉科技大学 | Antibacterial and anti-inflammatory dendrobium toothpaste |
| CN115636883A (en) * | 2022-09-05 | 2023-01-24 | 溯玄(上海)生物技术有限公司 | Plant purification component, preparation method, skin repair activity and application of plant purification component in preparation of skin repair cosmetics |
| CN115636883B (en) * | 2022-09-05 | 2024-04-26 | 广州梵之容化妆品有限公司 | Plant purification component, preparation method, skin repair activity and application of plant purification component in preparation of skin repair cosmetics |
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