CN104127509A - Method for extracting total anthraquinone from Rhizoma Polygoni Cuspidati through aqueous enzymatic process-semi-bionic process - Google Patents
Method for extracting total anthraquinone from Rhizoma Polygoni Cuspidati through aqueous enzymatic process-semi-bionic process Download PDFInfo
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Abstract
The invention discloses a method for extracting total anthraquinone from from Rhizoma Polygoni Cuspidati through an aqueous enzymatic process-semi-bionic process. The method comprises the following steps: 1, weighing Rhizoma Polygoni Cuspidati powder, adding a hydrolase reaction liquid, adjusting to an appropriate pH value, uniformly stirring, carrying out enzymatic hydrolysis for 4-8h, carrying out high temperature enzyme inactivation, and filtering; 2, adding a citric acid-sodium dihydrogen phosphate buffer solution to the enzymatic hydrolyzed Rhizoma Polygoni Cuspidati in order to adjust the pH value to 1.5-3.0, extracting, and filtering; 3, adding the citric acid-sodium dihydrogen phosphate buffer solution to filter residues in order to adjust the pH value to 6.5-7.5, extracting, and filtering; 4, adding the citric acid-sodium dihydrogen phosphate buffer solution to newly obtained filter residues in order to adjust the pH value to 8.0-8.5, extracting, and filtering; and 5, mixing filtrates obtained in step 1-4, concentrating, and carrying out vacuum drying to obtain an extract containing Rhizoma Polygoni Cuspidati anthraquinone. The method avoids the use of organic solvents; compared with microwave extraction, ultrasonic extraction and other assisted extraction methods, the method disclosed in the invention has the advantages of high extraction rate, simple extraction device and mild extraction conditions; and the method disclosed in the invention is an environmentally-friendly extraction method suitable for industrialized production.
Description
Technical field
The invention belongs to natural product extracting technique field, relate to a kind of extracting method of anthraquinone, be specifically related to the method that a kind of aqueous enzymatic method-semi-biomimetic method extracts general anthraquinone in Rhizoma Polygoni Cuspidati.
Background technology
Rhizoma Polygoni Cuspidati is root and the rhizome of Polygonaceae Liao platymiscium Rhizoma Polygoni Cuspidati, is China's Chinese medicine, and bitter in the mouth is cold in nature, the effects such as the analgesic therapy of invigorating blood circulation, clearing away heat-damp and promoting diuresis, preventing phlegm from forming and stopping coughing.Clinical in diseases such as jaundice due to damp-heat, cough due to lung-heat, sore, carbuncle and painful swelling, traumatic injury.In Rhizoma Polygoni Cuspidati, mainly contain Anthraquinones, stilbene class, flavone, liposoluble ingredient, the important physiologically active such as that these compositions have is antibacterial, antithrombotic, blood fat reducing.Anthraquinones of Polygonum Cuspidatum Sieb is its important antimicrobial component, and it extracts the organic solvent extraction that adopt more, and cost is high, environmental pollution is large, and also useful microwave extraction, supersound extraction, high to equipment requirements.Therefore, to take the eco-friendly extracting mode that water is solvent significant in exploitation.
Aqueous enzymatic method is a kind of extracting method of comparatively high-efficiency environment friendly, but extraction efficiency is not high, has caused to a certain extent the waste of herb resource.Semi-bionic extraction is that aids drug enters gastrointestinal tract environment, from a kind of new extraction process of biopharmaceutical angle design.
Summary of the invention
The object of the present invention is to provide a kind of aqueous enzymatic method-semi-biomimetic method to extract the method for general anthraquinone in Rhizoma Polygoni Cuspidati, the method combines aqueous enzymatic method for the extraction of Rhizoma Polygoni Cuspidati general anthraquinone with semi-biomimetic method, can significantly improve extraction efficiency, simple to operate, low for equipment requirements, be applicable to the demand that large-scale industrialization is produced.
The present invention is achieved through the following technical solutions:
Aqueous enzymatic method-semi-biomimetic method extracts a method for general anthraquinone in Rhizoma Polygoni Cuspidati, comprises the following steps:
1) first, get Giant Knotweed Rhizome, by 1g:(5～20) solid-liquid ratio of mL adds hydrolytic enzyme reactant liquor in Giant Knotweed Rhizome, fully mix, regulate pH value to 4.0～5.5, then under the stirring condition of 45～55 ℃, enzymolysis 4～8 hours, then process through high-temperature inactivation enzyme, finally filter;
Wherein, every 100mL hydrolytic enzyme reactant liquor is according to adding the ratio of 125mg cellulase that cellulase is dissolved in distilled water and is made in every 100mL distilled water;
2) to step 1) add citric acid-phosphate sodium dihydrogen buffer solution in filtering residue after filtering, fully mix, regulate pH value to 1.5～3.0, water-bath is filtered after extracting;
3) to step 2) add citric acid-phosphate sodium dihydrogen buffer solution in filtering residue after filtering, fully mix, regulate pH value to 6.5～7.5, water-bath is filtered after extracting;
4) to step 3) add citric acid-phosphate sodium dihydrogen buffer solution in filtering residue after filtering, fully mix, regulate pH value to 8.0～8.5, water-bath is filtered after extracting;
5) by step 1)～4) filter the filtrate obtain and merge, dried after solvent reclaimed, the dry extract that obtaining of obtaining comprises Rhizoma Polygoni Cuspidati total anthraquinones.
Step 1) described high-temperature inactivation enzyme processing is that the bath temperature of the reaction system after enzymolysis processing is increased to 80 ℃, heating 10min.
Step 2), 3), 4), the reaction material liquor ratio of filtering residue and citric acid-phosphate sodium dihydrogen buffer solution is 1g:(10～22) mL.
Step 2), 3), 4) described water-bath extraction temperature is 50～80 ℃.
Step 2), 3), 4) described water-bath extraction time is 1～3h.
Step 5) after described recovery solvent, dried is at 60～80 ℃, to carry out vacuum drying treatment after the filtrate decompression after merging is concentrated.
Described Giant Knotweed Rhizome is, after Rhizoma Polygoni Cuspidati is cleaned up, through pulverizing and sieving, drying, to constant weight, to make.
Compared with prior art, the present invention has following useful technique effect:
The present invention adopts aqueous enzymatic method from Rhizoma Polygoni Cuspidati, to extract general anthraquinone in conjunction with the method for semi-biomimetic method, and the extracting method adopting all not with an organic solvent, has been avoided problem of environmental pollution and reduced running cost.In leaching process, first add water enzyme digestion reaction liquid to carry out enzymolysis processing, this water enzyme digestion reaction liquid is can clear up in the cellulase solution distilled water of plant cell wall to make, stirring condition under carry out enzymolysis, can decompose to greatest extent destruction to the cell wall of Rhizoma Polygoni Cuspidati, thereby reduced resistance to mass tranfer, be more conducive to the stripping of effective ingredient.And when enzymolysis, regulator solution pH value is to the optimum pH (4.0～5.5) of enzyme, and adopts optimal reactive temperature (45～55 ℃), is beneficial to enzyme and plays effectiveness.The extraction that is more conducive to general anthraquinone after enzymolysis, has improved extraction efficiency.At semi-biomimetic method, in leaching process, meet the feature of Chinese medicine clinical practice, the process that simulation oral drugs absorb in gastrointestinal tract transhipment.The present invention adopts citric acid-sodium hydrogen phosphate buffer system as the extraction solvent of semi-biomimetic method, by different pH value gradients is set, reaches the object that aids drug is transported absorption process in vivo.
The aqueous enzymatic method that the present invention adopts-semi-biomimetic method extracts the general anthraquinone in Rhizoma Polygoni Cuspidati, follows the example of with microwave-assisted extraction method and compares with ultrasonic assisted Extraction, and extraction ratio is significantly improved, and to equipment require low.Extraction conditions of the present invention is gentle, with low cost, environmental friendliness, be applicable to large-scale industrial production, has a good application prospect.
The specific embodiment
Below in conjunction with specific embodiment, the present invention is described in further detail, and the explanation of the invention is not limited.
Embodiment 1
Aqueous enzymatic method-semi-biomimetic method extracts a method for general anthraquinone in Rhizoma Polygoni Cuspidati, comprises the following steps:
1) processing of Rhizoma Polygoni Cuspidati: Rhizoma Polygoni Cuspidati cleaning-drying, shatter and sieve, dry to constant weight, obtain Rhizoma Polygoni Cuspidati powder;
2) aqueous enzymatic extraction: accurately take 10g Rhizoma Polygoni Cuspidati powder, by the solid-liquid ratio of 1g:16mL, in Giant Knotweed Rhizome, add hydrolytic enzyme reactant liquor, after mixing, regulate pH value to 4.5, under stirring condition, in 50 ℃ of water-baths, enzymolysis is 6 hours, rising bath temperature to 80 ℃, heat cold filtration 10 minutes; Wherein, described hydrolytic enzyme reactant liquor is according to adding the ratio of 125mg cellulase that cellulase is dissolved in distilled water and is made in every 100mL distilled water;
3) in filtering residue semi-biomimetic method extraction: according to the solid-liquid ratio of solid-liquid ratio 1g:18mL, to step 1) filtering, add citric acid-phosphate sodium dihydrogen buffer solution, regulating pH value is 2.0, stirs, and in 70 ℃ of water-baths, extracts after 2h, filters; It is to filter after 2h is extracted in 7.5,70 ℃ of water-baths that filtering residue adds citric acid-phosphate sodium dihydrogen buffer solution to regulate pH value according to above solid-liquid ratio; It is to filter after 2h is extracted in 8.0,70 ℃ of water-baths that filtering residue adds citric acid-phosphate sodium dihydrogen buffer solution to regulate pH value according to above solid-liquid ratio again;
4) by step 2), 3) in filter all filtrates obtain and merge, on Rotary Evaporators, be concentrated into fluid extract, then in vacuum drying oven with 60 ℃ of vacuum dryings, obtain the dry extract 7.08g that contains Anthraquinones of Polygonum Cuspidatum Sieb.
Emodin content in Anthraquinones of Polygonum Cuspidatum Sieb dry extract is measured:
1, the foundation of emodin standard curve
Precision takes 10mg emodin standard substance to 100mL volumetric flask, adds methanol: 0.015mol/L phosphate aqueous solution (80:20) is settled to scale as solvent, shakes up.Precision measures in above-mentioned standard solution 0.2mL, 0.4mL, 0.6mL, 0.8mL, 1.0mL, 1.2mL to a six 10mL volumetric flask respectively, add solvent to be settled to scale, shake up, obtain emodin solution concentration and be respectively 0.002mg/mL, 0.004mg/mL, 0.006mg/mL, 0.008mg/mL, 0.010mg/mL, 0.012mg/mL.With 0.25 μ m filtering with microporous membrane, difference sample introduction 20 μ L, each concentration sample introduction 3 times.Mobile phase is methanol: 0.015mol/L phosphate aqueous solution (80:20), and isocratic elution, ultraviolet detection wavelength is 284nm, and chromatographic column is Diamonsil C18 (5 μ m, 250 * 4.6mm), and flow velocity is 1.0mL/min.Take average peak area value as vertical coordinate, and emodin concentration is abscissa, drawing standard curve, and acquisition linear equation is Y=254155.29X-5173.93 (R2=0.9997), concentration range is 0.002～0.012mg/mL.
2, in extract, emodin content is measured
Precision takes the above-mentioned dry extract of 10mg, in 10mL volumetric flask, adds mobile phase to make solvent and is settled to scale, shakes up, and filters sample introduction 20 μ L, sample introduction 3 times, averaged.Recording emodin content in dry extract is 6.95mg/10g medical material.
3, methodological study
1) precision: precision measures 0.1mg/mL emodin standard solution 1mL, is placed in 10mL volumetric flask, adds mobile phase solution and is settled to scale as solvent, shakes up, and filters, and sample introduction 20 μ L, repeat sample introduction 6 times.Recording relative standard deviation (RSD) is 1.27%.
2) repeatability: get said extracted extractum 100.32mg, be placed in 25mL volumetric flask, add mobile phase solution and be settled to scale, shake up.Precision measures the above-mentioned solution of 2mL to 10mL volumetric flask again, adds mobile phase solution and is settled to scale, shakes up, and filters, and sample introduction 20 μ L, repeat sample introduction 5 times.Recording relative standard deviation is 1.73%.
3) recovery of standard addition: precision takes said extracted extractum 199.78mg, adds respectively the emodin standard substance of emodin content 80%, 100% wherein and 120%, is placed in 25mL volumetric flask, adds mobile phase solution and is settled to scale, shakes up.Precision measures this solution of 5mL and is placed in 50mL volumetric flask respectively again, adds mobile phase solution and is settled to scale, shakes up, and filters.Sample introduction 20 μ L.According to following formula, calculate recovery of standard addition: recovery of standard addition=(mark-on Specimen Determination value-Specimen Determination value)/add scalar * 100%.It is 101.38% that final calculating obtains recovery of standard addition, and relative standard deviation is 2.97%.
Embodiment 2
Aqueous enzymatic method-semi-biomimetic method extracts a method for general anthraquinone in Rhizoma Polygoni Cuspidati, comprises the following steps:
1) processing of Rhizoma Polygoni Cuspidati: Rhizoma Polygoni Cuspidati cleaning-drying, shatter and sieve, dry to constant weight, obtain Rhizoma Polygoni Cuspidati powder;
2) aqueous enzymatic extraction: accurately take 10g Rhizoma Polygoni Cuspidati powder, by the solid-liquid ratio of 1g:10mL, in Giant Knotweed Rhizome, add hydrolytic enzyme reactant liquor, after mixing, regulate pH value to 4.5, under stirring condition, in 50 ℃ of water-baths, enzymolysis is 7 hours, rising bath temperature to 80 ℃, heat cold filtration 10 minutes; Wherein, described hydrolytic enzyme reactant liquor is according to adding the ratio of 125mg cellulase that cellulase is dissolved in distilled water and is made in every 100mL distilled water;
3) in filtering residue semi-biomimetic method extraction: according to the solid-liquid ratio of solid-liquid ratio 1g:14mL, to step 1) filtering, add citric acid-phosphate sodium dihydrogen buffer solution, regulating pH value is 2.0, stirs, and in 80 ℃ of water-baths, extracts after 2h, filters; It is to filter after 2h is extracted in 7.5,80 ℃ of water-baths that filtering residue adds citric acid-phosphate sodium dihydrogen buffer solution to regulate pH value according to above solid-liquid ratio; It is to filter after 2h is extracted in 8.0,80 ℃ of water-baths that filtering residue adds citric acid-phosphate sodium dihydrogen buffer solution to regulate pH value according to above solid-liquid ratio again;
4) by step 2), 3) in filter all filtrates obtain and merge, on Rotary Evaporators, be concentrated into fluid extract, then in vacuum drying oven with 60 ℃ of vacuum dryings, obtain the dry extract 7.02g that contains Anthraquinones of Polygonum Cuspidatum Sieb.
Emodin content in extract is measured: precision takes the above-mentioned dry extract of 10mg, in 10mL volumetric flask, adds mobile phase to make solvent and is settled to scale, shaken up, filter sample introduction 20 μ L, sample introduction 3 times, averaged.Recording emodin content in dry extract is 3.87mg/10g medical material.
Embodiment 3
Aqueous enzymatic method-semi-biomimetic method extracts a method for general anthraquinone in Rhizoma Polygoni Cuspidati, comprises the following steps:
1) processing of Rhizoma Polygoni Cuspidati: Rhizoma Polygoni Cuspidati cleaning-drying, shatter and sieve, dry to constant weight, obtain Rhizoma Polygoni Cuspidati powder;
2) aqueous enzymatic extraction: accurately take 10g Rhizoma Polygoni Cuspidati powder, by the solid-liquid ratio of 1g:15mL, in Giant Knotweed Rhizome, add hydrolytic enzyme reactant liquor, after mixing, regulate pH value to 4.0, under stirring condition, in 55 ℃ of water-baths, enzymolysis is 6 hours, rising bath temperature to 80 ℃, heat cold filtration 10 minutes; Wherein, every 100mL hydrolytic enzyme reactant liquor is according to adding the ratio of 125mg cellulase that cellulase is dissolved in distilled water and is made in every 100mL distilled water;
3) in filtering residue semi-biomimetic method extraction: according to the solid-liquid ratio of solid-liquid ratio 1g:22mL, to step 1) filtering, add citric acid-phosphate sodium dihydrogen buffer solution, regulating pH value is 2.0, stirs, and in 70 ℃ of water-baths, extracts after 1.5h, filters; It is to filter after 1.5h is extracted in 7.5,70 ℃ of water-baths that filtering residue adds citric acid-phosphate sodium dihydrogen buffer solution to regulate pH value according to above solid-liquid ratio; It is to filter after 1.5h is extracted in 8.0,70 ℃ of water-baths that filtering residue adds citric acid-phosphate sodium dihydrogen buffer solution to regulate pH value according to above solid-liquid ratio again;
4) by step 2), 3) in filter all filtrates obtain and merge, on Rotary Evaporators, be concentrated into fluid extract, then in vacuum drying oven with 60 ℃ of vacuum dryings, obtain the dry extract 7.83g that contains Anthraquinones of Polygonum Cuspidatum Sieb.
Emodin content in extract is measured: precision takes the above-mentioned dry extract of 10mg, in 10mL volumetric flask, adds mobile phase to make solvent and is settled to scale, shaken up, filter sample introduction 20 μ L, sample introduction 3 times, averaged.Recording emodin content in dry extract is 3.34mg/10g medical material.
Embodiment 4
Aqueous enzymatic method-semi-biomimetic method extracts a method for general anthraquinone in Rhizoma Polygoni Cuspidati, comprises the following steps:
1) processing of Rhizoma Polygoni Cuspidati: Rhizoma Polygoni Cuspidati cleaning-drying, shatter and sieve, dry to constant weight, obtain Rhizoma Polygoni Cuspidati powder;
2) aqueous enzymatic extraction: accurately take 10g Rhizoma Polygoni Cuspidati powder, by the solid-liquid ratio of 1g:5mL, in Giant Knotweed Rhizome, add hydrolytic enzyme reactant liquor, after mixing, regulate pH value to 5.0, under stirring condition, in 45 ℃ of water-baths, enzymolysis is 8 hours, rising bath temperature to 80 ℃, heat cold filtration 10 minutes; Wherein, every 100mL hydrolytic enzyme reactant liquor is according to adding the ratio of 125mg cellulase that cellulase is dissolved in distilled water and is made in every 100mL distilled water;
3) in filtering residue semi-biomimetic method extraction: according to the solid-liquid ratio of solid-liquid ratio 1g:10mL, to step 1) filtering, add citric acid-phosphate sodium dihydrogen buffer solution, regulating pH value is 1.5, stirs, and in 50 ℃ of water-baths, extracts after 3h, filters; It is to filter after 3h is extracted in 6.5,50 ℃ of water-baths that filtering residue adds citric acid-phosphate sodium dihydrogen buffer solution to regulate pH value according to above solid-liquid ratio; It is to filter after 3h is extracted in 8.3,50 ℃ of water-baths that filtering residue adds citric acid-phosphate sodium dihydrogen buffer solution to regulate pH value according to above solid-liquid ratio again;
4) by step 2), 3) in filter all filtrates obtain and merge, on Rotary Evaporators, be concentrated into fluid extract, then in vacuum drying oven with 70 ℃ of vacuum dryings, obtain the dry extract 7.57g that contains Anthraquinones of Polygonum Cuspidatum Sieb.
Embodiment 5
Aqueous enzymatic method-semi-biomimetic method extracts a method for general anthraquinone in Rhizoma Polygoni Cuspidati, comprises the following steps:
1) processing of Rhizoma Polygoni Cuspidati: Rhizoma Polygoni Cuspidati cleaning-drying, shatter and sieve, dry to constant weight, obtain Rhizoma Polygoni Cuspidati powder;
2) aqueous enzymatic extraction: accurately take 10g Rhizoma Polygoni Cuspidati powder, by the solid-liquid ratio of 1g:20mL, in Giant Knotweed Rhizome, add hydrolytic enzyme reactant liquor, after mixing, regulate pH value to 5.5, under stirring condition, in 55 ℃ of water-baths, enzymolysis is 4 hours, rising bath temperature to 80 ℃, heat cold filtration 10 minutes; Wherein, every 100mL hydrolytic enzyme reactant liquor is according to adding the ratio of 125mg cellulase that cellulase is dissolved in distilled water and is made in every 100mL distilled water;
3) in filtering residue semi-biomimetic method extraction: according to the solid-liquid ratio of solid-liquid ratio 1g:15mL, to step 1) filtering, add citric acid-phosphate sodium dihydrogen buffer solution, regulating pH value is 3.0, stirs, and in 80 ℃ of water-baths, extracts after 1h, filters; It is to filter after 1h is extracted in 7.2,80 ℃ of water-baths that filtering residue adds citric acid-phosphate sodium dihydrogen buffer solution to regulate pH value according to above solid-liquid ratio; It is to filter after 1h is extracted in 8.5,80 ℃ of water-baths that filtering residue adds citric acid-phosphate sodium dihydrogen buffer solution to regulate pH value according to above solid-liquid ratio again;
4) by step 2), 3) in filter all filtrates obtain and merge, on Rotary Evaporators, be concentrated into fluid extract, then in vacuum drying oven with 80 ℃ of vacuum dryings, obtain the dry extract 7.24g that contains Anthraquinones of Polygonum Cuspidatum Sieb.
Comparative example 1
Ultrasonic assisted Extraction is followed the example of the method for extracting general anthraquinone in Rhizoma Polygoni Cuspidati, comprises the following steps:
(1) processing of Rhizoma Polygoni Cuspidati: Rhizoma Polygoni Cuspidati cleaning-drying, shatter and sieve, dry to constant weight, obtain Rhizoma Polygoni Cuspidati powder.
(2) ultrasonic assisted extraction: accurately take three parts of each 10g of Rhizoma Polygoni Cuspidati powder, 1:40 adds distilled water according to solid-liquid ratio, and 50 ℃ of preheatings, after 20 minutes, adopt ultrasonic auxiliary extraction, and ultrasonic power is 10kW/m3, and dutycycle is 2:1, and ultrasonic time is 1 hour.Repeat to extract merging filtrate 3 times.
(3) make extractum: the filtrate of extracting for 3 times merges respectively, is concentrated into fluid extract on Rotary Evaporators, then in vacuum drying oven with 60 ℃ of vacuum dryings, the average quality that obtains the dry extract that contains Anthraquinones of Polygonum Cuspidatum Sieb is 3.31g.
(4) in extract, emodin content is measured: precision takes the above-mentioned dry extract of 10mg respectively, in 10mL volumetric flask, adds mobile phase to make solvent and is settled to scale, shakes up, and filters sample introduction 20 μ L, sample introduction 3 times, averaged.Recording emodin content in dry extract is 1.43mg/10g medical material.
Comparative example 2
Microwave-assisted extraction method is extracted the method for general anthraquinone in Rhizoma Polygoni Cuspidati, comprises the following steps:
(1) processing of Rhizoma Polygoni Cuspidati: Rhizoma Polygoni Cuspidati cleaning-drying, shatter and sieve, dry to constant weight, obtain Rhizoma Polygoni Cuspidati powder.
(2) microwave-assisted extracts: accurately take three parts of each 10g of Rhizoma Polygoni Cuspidati powder, according to solid-liquid ratio, 1:40 adds distilled water, and 70 ℃ of preheatings, after 20 minutes, adopt microwave auxiliary extraction, and microwave power is 480W, and dutycycle is 2:3, and ultrasonic time is 20 minutes.Repeat to extract merging filtrate 3 times.
(3) make extractum: the filtrate of extracting for 3 times merges respectively, is concentrated into fluid extract on Rotary Evaporators, then in vacuum drying oven with 60 ℃ of vacuum dryings, the average quality that obtains the dry extract that contains Anthraquinones of Polygonum Cuspidatum Sieb is 5.04g.
(4) in extract, emodin content is measured: precision takes the above-mentioned dry extract of 10mg respectively, in 10mL volumetric flask, adds mobile phase to make solvent and is settled to scale, shakes up, and filters sample introduction 20 μ L, sample introduction 3 times, averaged.Recording emodin content in dry extract is 1.97mg/10g medical material.
From embodiments of the invention, compare with comparative example, can find out: the yield of extract that aqueous enzymatic method-semi-biomimetic method extracts the acquisition of Rhizoma Polygoni Cuspidati powder is the highest, and in extractum, the content of emodin is also the highest.In the application of enzyme and leaching process, the gradient of pH value raises and all can promote the stripping of active substance, thereby the new method for extracting of Rhizoma Polygoni Cuspidati provided by the present invention has certain superiority.
Claims (7)
1. aqueous enzymatic method-semi-biomimetic method extracts a method for general anthraquinone in Rhizoma Polygoni Cuspidati, it is characterized in that, comprises the following steps:
1) first, get Giant Knotweed Rhizome, by 1g:(5～20) solid-liquid ratio of mL adds hydrolytic enzyme reactant liquor in Giant Knotweed Rhizome, fully mix, regulate pH value to 4.0～5.5, then under the stirring condition of 45～55 ℃, enzymolysis 4～8 hours, then process through high-temperature inactivation enzyme, finally filter;
Wherein, every 100mL hydrolytic enzyme reactant liquor is according to adding the ratio of 125mg cellulase that cellulase is dissolved in distilled water and is made in every 100mL distilled water;
2) to step 1) add citric acid-phosphate sodium dihydrogen buffer solution in filtering residue after filtering, fully mix, regulate pH value to 1.5～3.0, water-bath is filtered after extracting;
3) to step 2) add citric acid-phosphate sodium dihydrogen buffer solution in filtering residue after filtering, fully mix, regulate pH value to 6.5～7.5, water-bath is filtered after extracting;
4) to step 3) add citric acid-phosphate sodium dihydrogen buffer solution in filtering residue after filtering, fully mix, regulate pH value to 8.0～8.5, water-bath is filtered after extracting;
5) by step 1)～4) filter the filtrate obtain and merge, dried after solvent reclaimed, the dry extract that obtaining of obtaining comprises Rhizoma Polygoni Cuspidati total anthraquinones.
2. a kind of aqueous enzymatic method according to claim 1-semi-biomimetic method extracts the method for general anthraquinone in Rhizoma Polygoni Cuspidati, it is characterized in that step 1) described high-temperature inactivation enzyme is processed is that the bath temperature of the reaction system after enzymolysis processing is increased to 80 ℃, heating 10min.
3. a kind of aqueous enzymatic method according to claim 1-semi-biomimetic method extracts the method for general anthraquinone in Rhizoma Polygoni Cuspidati, it is characterized in that step 2), 3), 4) in the reaction material liquor ratio of filtering residue and citric acid-phosphate sodium dihydrogen buffer solution be 1g:(10～22) mL.
4. a kind of aqueous enzymatic method according to claim 1-semi-biomimetic method extracts the method for general anthraquinone in Rhizoma Polygoni Cuspidati, it is characterized in that step 2), 3), 4) to extract temperature be 50～80 ℃ in described water-bath.
5. a kind of aqueous enzymatic method according to claim 1-semi-biomimetic method extracts the method for general anthraquinone in Rhizoma Polygoni Cuspidati, it is characterized in that step 2), 3), 4) described water-bath extraction time is 1～3h.
6. a kind of aqueous enzymatic method according to claim 1-semi-biomimetic method extracts the method for general anthraquinone in Rhizoma Polygoni Cuspidati, it is characterized in that step 5) dried is at 60～80 ℃, to carry out vacuum drying treatment after the filtrate decompression after merging is concentrated after described recovery solvent.
7. according to a kind of aqueous enzymatic method described in any one in claim 1～6-semi-biomimetic method, extract the method for general anthraquinone in Rhizoma Polygoni Cuspidati, it is characterized in that, described Giant Knotweed Rhizome is, after Rhizoma Polygoni Cuspidati is cleaned up, through pulverizing and sieving, drying, to constant weight, to make.
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| CN103202884A (en) * | 2013-04-03 | 2013-07-17 | 西安交通大学 | Method for extracting hovenaia dulcis total alkaloid by semi-bionic-enzymic method |
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| CN106188331A (en) * | 2016-08-12 | 2016-12-07 | 黑龙江众生生物工程有限公司 | The method that a kind of biomimetic method associating enzymatic isolation method extracts Auricularia polycose |
| 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 |
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