CN104127509B - A kind of aqueous enzymatic method-semi-biomimetic method extracts the method for general anthraquinone in Rhizoma Polygoni Cuspidati - Google Patents
A kind of aqueous enzymatic method-semi-biomimetic method extracts the method for general anthraquinone in Rhizoma Polygoni Cuspidati Download PDFInfo
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Abstract
The invention discloses the method that a kind of aqueous enzymatic method-semi-biomimetic method extracts general anthraquinone in Rhizoma Polygoni Cuspidati, comprise the following steps: 1) claim Rhizoma Polygoni Cuspidati powder, add hydrolytic enzyme reactant liquor, and be adjusted to optimum pH, stir, high-temperature inactivation enzyme after enzymolysis 4 ~ 8h, filters; 2) adding citrate-phosphate sodium dihydrogen buffer adjust ph in the Rhizoma Polygoni Cuspidati after enzymolysis is 1.5 ~ 3.0, filters after extracting; 3) filtering residue adds citrate-phosphate sodium dihydrogen buffer adjust ph is 6.5 ~ 7.5, filters after extracting; 4) filtering residue adds citrate-phosphate sodium dihydrogen buffer adjust ph is 8.0 ~ 8.5, filters after extracting; 5) 1 is merged) ~ 4) filtrate, concentrated also vacuum drying, obtains the extract containing Anthraquinones of Polygonum Cuspidatum Sieb.This method is avoided with an organic solvent, and with other assisted extraction methods, as microwave extraction, supersound extraction are compared, extraction ratio is high, and extraction equipment is simple, and extraction conditions is gentle, is a kind of environmental friendliness, the extracting method that suitability for industrialized is produced.
Description
Technical field
The invention belongs to technical field of natural product extraction, 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 the root welding technology of Polygonaceae Liao platymiscium Rhizoma Polygoni Cuspidati, and be China's Chinese medicine, bitter in the mouth is cold in nature, the effects such as 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.Main containing Anthraquinones, stilbene class, flavone, liposoluble ingredient in Rhizoma Polygoni Cuspidati, these compositions have the important physiologically actives such as antibacterial, antithrombotic, blood fat reducing.Anthraquinones of Polygonum Cuspidatum Sieb is its important antimicrobial component, and its extraction adopts organic solvent extraction more, and cost is high, environmental pollution is large, and also useful microwave extraction, supersound extraction, high to equipment requirements.Therefore, develop with water is that the eco-friendly extracting mode of solvent is significant.
Aqueous enzymatic method is a kind of extracting method of comparatively high-efficiency environment friendly, but extraction efficiency is not high, causes the waste of herb resource to a certain extent.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
A kind of aqueous enzymatic method-semi-biomimetic method is the object of the present invention is to provide to extract the method for general anthraquinone in Rhizoma Polygoni Cuspidati, aqueous enzymatic method combines with semi-biomimetic method for the extraction of general anthraquinone in Rhizoma Polygoni Cuspidati by the method, extraction efficiency can be significantly improved, simple to operate, low for equipment requirements, be applicable to the demand of industrialization large-scale production.
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, abundant mixing, adjust ph to 4.0 ~ 5.5, then under the stirring condition of 45 ~ 55 DEG C, enzymolysis 4 ~ 8 hours, again through high-temperature inactivation ferment treatment, finally filter;
Wherein, every 100mL hydrolytic enzyme reactant liquor to be dissolved in distilled water by cellulase according to the ratio adding 125mg cellulase in every 100mL distilled water to make;
2) to step 1) filter after filtering residue in add citrate-phosphate sodium dihydrogen buffer, fully mix, adjust ph to 1.5 ~ 3.0, water-bath extract after filter;
3) to step 2) filter after filtering residue in add citrate-phosphate sodium dihydrogen buffer, fully mix, adjust ph to 6.5 ~ 7.5, water-bath extract after filter;
4) to step 3) filter after filtering residue in add citrate-phosphate sodium dihydrogen buffer, fully mix, adjust ph to 8.0 ~ 8.5, water-bath extract after filter;
5) by step 1) ~ 4) filter the filtrate merging obtained, dried after recycling design, the dry extract obtaining comprising Rhizoma Polygoni Cuspidati total anthraquinones obtained.
Step 1) described in high-temperature inactivation ferment treatment be by enzymolysis processing after the bath temperature of reaction system be increased to 80 DEG C, heating 10min.
Step 2), 3), 4) in the reaction material liquor ratio of filtering residue and citrate-phosphate sodium dihydrogen buffer be 1g:(10 ~ 22) mL.
Step 2), 3), 4) described in water-bath Extracting temperature be 50 ~ 80 DEG C.
Step 2), 3), 4) described in water-bath extraction time be 1 ~ 3h.
Step 5) described in recycling design after dried be by merge after filtrate reduced in volume after at 60 ~ 80 DEG C, carry out vacuum drying treatment.
Described Giant Knotweed Rhizome is after being cleaned up by Rhizoma Polygoni Cuspidati, obtained through pulverizing and sieving, drying to constant weight.
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 adopted all not with an organic solvent, avoids problem of environmental pollution and reduces running cost.In leaching process, first add water enzyme digestion reaction liquid carry out enzymolysis processing, this water enzyme digestion reaction liquid makes in the cellulase solution distilled water can clearing up plant cell wall, stirring condition under carry out enzymolysis, destruction can be decomposed to greatest extent to the cell wall of Rhizoma Polygoni Cuspidati, thus reduce resistance to mass tranfer, be more conducive to the stripping of effective ingredient.And regulate solution ph to the optimum pH (4.0 ~ 5.5) of enzyme when enzymolysis, and adopt optimal reactive temperature (45 ~ 55 DEG C), be beneficial to enzyme and play effectiveness.Be more conducive to the extraction of general anthraquinone after enzymolysis, improve extraction efficiency.In leaching process, the feature of Chinese medicine clinical practice is met, the process that simulation oral drugs absorb at gastrointestinal transit at semi-biomimetic method.The present invention adopts citrate-phosphate disodium hydrogen buffer system as the Extraction solvent of semi-biomimetic method, by arranging different pH value gradients, reaches the object that aids drug transports absorption process in vivo.
The aqueous enzymatic method that the present invention adopts-semi-biomimetic method extracts the general anthraquinone in Rhizoma Polygoni Cuspidati, and compare with microwave―assisted extraction with ultrasound assisted extraction method, extraction ratio is significantly improved, and low to the requirement of equipment.Extraction conditions of the present invention is gentle, with low cost, environmental friendliness, applicable large-scale industrial production, has a good application prospect.
Detailed description of the invention
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) process of Rhizoma Polygoni Cuspidati: Rhizoma Polygoni Cuspidati cleaning-drying, shatters and sieve, dries to constant weight, obtains Rhizoma Polygoni Cuspidati powder;
2) aqueous enzymatic extraction: accurately take 10g Rhizoma Polygoni Cuspidati powder, in Giant Knotweed Rhizome, hydrolytic enzyme reactant liquor is added by the solid-liquid ratio of 1g:16mL, after mixing, adjust ph to 4.5, under agitation, enzymolysis 6 hours in 50 DEG C of water-baths, raises bath temperature to 80 DEG C, heat 10 minutes, cold filtration; Wherein, described hydrolytic enzyme reactant liquor to be dissolved in distilled water by cellulase according to the ratio adding 125mg cellulase in every 100mL distilled water to make;
3) semi-biomimetic method extracts: according to the solid-liquid ratio of solid-liquid ratio 1g:18mL, to step 1) add citrate-phosphate sodium dihydrogen buffer in the filtering residue that filters, adjust ph is 2.0, stirs, and after extracting 2h, filters in 70 DEG C of water-baths; It is filter after 2h is extracted in 7.5,70 DEG C of water-baths that filtering residue adds citrate-phosphate sodium dihydrogen buffer adjust ph according to above solid-liquid ratio; It is filter after 2h is extracted in 8.0,70 DEG C of water-baths that filtering residue adds citrate-phosphate sodium dihydrogen buffer adjust ph according to above solid-liquid ratio again;
4) by step 2), 3) in filter all filtrates obtained and merge, be concentrated into fluid extract on a rotary evaporator, then in vacuum drying oven with 60 DEG C of vacuum dryings, obtain the dry extract 7.08g containing 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 in 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 and 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 μm of filtering with microporous membrane, sample introduction 20 μ L respectively, each concentration sample introduction 3 times.Mobile phase is methanol: 0.015mol/L phosphate aqueous solution (80:20), isocratic elution, and ultraviolet detection wavelength is 284nm, and chromatographic column is DiamonsilC18 (5 μm, 250 × 4.6mm), and flow velocity is 1.0mL/min.With average peak area value for vertical coordinate, emodin concentration is abscissa, drawing standard curve, and acquisition linear equation is Y=254155.29X-5173.93 (R2=0.9997), and concentration range is 0.002 ~ 0.012mg/mL.
2, in extract, emodin content measures
Precision takes the above-mentioned dry extract of 10mg, as in 10mL volumetric flask, adds mobile phase and makees solvent and be settled to scale, shake up, filter, 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, shake up, filters, sample introduction 20 μ L, repeats 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 in 10mL volumetric flask again, adds mobile phase solution and is settled to scale, shake up, and filters, sample introduction 20 μ L, repeats sample introduction 5 times.Recording relative standard deviation is 1.73%.
3) recovery of standard addition: precision takes said extracted extractum 199.78mg, adds the emodin standard substance of wherein emodin content 80%, 100% and 120% respectively, is placed in 25mL volumetric flask, adds mobile phase solution and is settled to scale, shake 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, shake up, and filters.Sample introduction 20 μ L.According to following formulae discovery 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) process of Rhizoma Polygoni Cuspidati: Rhizoma Polygoni Cuspidati cleaning-drying, shatters and sieve, dries to constant weight, obtains Rhizoma Polygoni Cuspidati powder;
2) aqueous enzymatic extraction: accurately take 10g Rhizoma Polygoni Cuspidati powder, in Giant Knotweed Rhizome, hydrolytic enzyme reactant liquor is added by the solid-liquid ratio of 1g:10mL, after mixing, adjust ph to 4.5, under agitation, enzymolysis 7 hours in 50 DEG C of water-baths, raises bath temperature to 80 DEG C, heat 10 minutes, cold filtration; Wherein, described hydrolytic enzyme reactant liquor to be dissolved in distilled water by cellulase according to the ratio adding 125mg cellulase in every 100mL distilled water to make;
3) semi-biomimetic method extracts: according to the solid-liquid ratio of solid-liquid ratio 1g:14mL, to step 1) add citrate-phosphate sodium dihydrogen buffer in the filtering residue that filters, adjust ph is 2.0, stirs, and after extracting 2h, filters in 80 DEG C of water-baths; It is filter after 2h is extracted in 7.5,80 DEG C of water-baths that filtering residue adds citrate-phosphate sodium dihydrogen buffer adjust ph according to above solid-liquid ratio; It is filter after 2h is extracted in 8.0,80 DEG C of water-baths that filtering residue adds citrate-phosphate sodium dihydrogen buffer adjust ph according to above solid-liquid ratio again;
4) by step 2), 3) in filter all filtrates obtained and merge, be concentrated into fluid extract on a rotary evaporator, then in vacuum drying oven with 60 DEG C of vacuum dryings, obtain the dry extract 7.02g containing Anthraquinones of Polygonum Cuspidatum Sieb.
Emodin content in extract is measured: precision takes the above-mentioned dry extract of 10mg, as in 10mL volumetric flask, add mobile phase and make solvent and be settled to scale, shake 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) process of Rhizoma Polygoni Cuspidati: Rhizoma Polygoni Cuspidati cleaning-drying, shatters and sieve, dries to constant weight, obtains Rhizoma Polygoni Cuspidati powder;
2) aqueous enzymatic extraction: accurately take 10g Rhizoma Polygoni Cuspidati powder, in Giant Knotweed Rhizome, hydrolytic enzyme reactant liquor is added by the solid-liquid ratio of 1g:15mL, after mixing, adjust ph to 4.0, under agitation, enzymolysis 6 hours in 55 DEG C of water-baths, raises bath temperature to 80 DEG C, heat 10 minutes, cold filtration; Wherein, every 100mL hydrolytic enzyme reactant liquor to be dissolved in distilled water by cellulase according to the ratio adding 125mg cellulase in every 100mL distilled water to make;
3) semi-biomimetic method extracts: according to the solid-liquid ratio of solid-liquid ratio 1g:22mL, to step 1) add citrate-phosphate sodium dihydrogen buffer in the filtering residue that filters, adjust ph is 2.0, stirs, and after extracting 1.5h, filters in 70 DEG C of water-baths; It is filter after 1.5h is extracted in 7.5,70 DEG C of water-baths that filtering residue adds citrate-phosphate sodium dihydrogen buffer adjust ph according to above solid-liquid ratio; It is filter after 1.5h is extracted in 8.0,70 DEG C of water-baths that filtering residue adds citrate-phosphate sodium dihydrogen buffer adjust ph according to above solid-liquid ratio again;
4) by step 2), 3) in filter all filtrates obtained and merge, be concentrated into fluid extract on a rotary evaporator, then in vacuum drying oven with 60 DEG C of vacuum dryings, obtain the dry extract 7.83g containing Anthraquinones of Polygonum Cuspidatum Sieb.
Emodin content in extract is measured: precision takes the above-mentioned dry extract of 10mg, as in 10mL volumetric flask, add mobile phase and make solvent and be settled to scale, shake 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) process of Rhizoma Polygoni Cuspidati: Rhizoma Polygoni Cuspidati cleaning-drying, shatters and sieve, dries to constant weight, obtains Rhizoma Polygoni Cuspidati powder;
2) aqueous enzymatic extraction: accurately take 10g Rhizoma Polygoni Cuspidati powder, in Giant Knotweed Rhizome, hydrolytic enzyme reactant liquor is added by the solid-liquid ratio of 1g:5mL, after mixing, adjust ph to 5.0, under agitation, enzymolysis 8 hours in 45 DEG C of water-baths, raises bath temperature to 80 DEG C, heat 10 minutes, cold filtration; Wherein, every 100mL hydrolytic enzyme reactant liquor to be dissolved in distilled water by cellulase according to the ratio adding 125mg cellulase in every 100mL distilled water to make;
3) semi-biomimetic method extracts: according to the solid-liquid ratio of solid-liquid ratio 1g:10mL, to step 1) add citrate-phosphate sodium dihydrogen buffer in the filtering residue that filters, adjust ph is 1.5, stirs, and after extracting 3h, filters in 50 DEG C of water-baths; It is filter after 3h is extracted in 6.5,50 DEG C of water-baths that filtering residue adds citrate-phosphate sodium dihydrogen buffer adjust ph according to above solid-liquid ratio; It is filter after 3h is extracted in 8.3,50 DEG C of water-baths that filtering residue adds citrate-phosphate sodium dihydrogen buffer adjust ph according to above solid-liquid ratio again;
4) by step 2), 3) in filter all filtrates obtained and merge, be concentrated into fluid extract on a rotary evaporator, then in vacuum drying oven with 70 DEG C of vacuum dryings, obtain the dry extract 7.57g containing 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) process of Rhizoma Polygoni Cuspidati: Rhizoma Polygoni Cuspidati cleaning-drying, shatters and sieve, dries to constant weight, obtains Rhizoma Polygoni Cuspidati powder;
2) aqueous enzymatic extraction: accurately take 10g Rhizoma Polygoni Cuspidati powder, in Giant Knotweed Rhizome, hydrolytic enzyme reactant liquor is added by the solid-liquid ratio of 1g:20mL, after mixing, adjust ph to 5.5, under agitation, enzymolysis 4 hours in 55 DEG C of water-baths, raises bath temperature to 80 DEG C, heat 10 minutes, cold filtration; Wherein, every 100mL hydrolytic enzyme reactant liquor to be dissolved in distilled water by cellulase according to the ratio adding 125mg cellulase in every 100mL distilled water to make;
3) semi-biomimetic method extracts: according to the solid-liquid ratio of solid-liquid ratio 1g:15mL, to step 1) add citrate-phosphate sodium dihydrogen buffer in the filtering residue that filters, adjust ph is 3.0, stirs, and after extracting 1h, filters in 80 DEG C of water-baths; It is filter after 1h is extracted in 7.2,80 DEG C of water-baths that filtering residue adds citrate-phosphate sodium dihydrogen buffer adjust ph according to above solid-liquid ratio; It is filter after 1h is extracted in 8.5,80 DEG C of water-baths that filtering residue adds citrate-phosphate sodium dihydrogen buffer adjust ph according to above solid-liquid ratio again;
4) by step 2), 3) in filter all filtrates obtained and merge, be concentrated into fluid extract on a rotary evaporator, then in vacuum drying oven with 80 DEG C of vacuum dryings, obtain the dry extract 7.24g containing Anthraquinones of Polygonum Cuspidatum Sieb.
Comparative example 1
Ultrasound assisted extraction method extracts the method for general anthraquinone in Rhizoma Polygoni Cuspidati, comprises the following steps:
(1) process of Rhizoma Polygoni Cuspidati: Rhizoma Polygoni Cuspidati cleaning-drying, shatters and sieve, dries to constant weight, obtains Rhizoma Polygoni Cuspidati powder.
(2) ultrasound assisted extraction: accurately take three parts of each 10g of Rhizoma Polygoni Cuspidati powder, add distilled water according to solid-liquid ratio 1:40,50 DEG C of preheatings are after 20 minutes, and adopt ultrasound on extracting, ultrasonic power is 10kW/m3, and dutycycle is 2:1, and ultrasonic time is 1 hour.Repeat extraction 3 times, merging filtrate.
(3) make extractum: the filtrate of extracting for 3 times merges respectively, is concentrated into fluid extract on a rotary evaporator, then in vacuum drying oven with 60 DEG C of vacuum dryings, the average quality of dry extract obtained containing Anthraquinones of Polygonum Cuspidatum Sieb is 3.31g.
(4) in extract, emodin content measures: precision takes the above-mentioned dry extract of 10mg respectively, as in 10mL volumetric flask, adds mobile phase and makees solvent and be settled to scale, shake up, filter, 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 extracts the method for general anthraquinone in Rhizoma Polygoni Cuspidati, comprises the following steps:
(1) process of Rhizoma Polygoni Cuspidati: Rhizoma Polygoni Cuspidati cleaning-drying, shatters and sieve, dries to constant weight, obtains Rhizoma Polygoni Cuspidati powder.
(2) microwave radiation exaraction: accurately take three parts of each 10g of Rhizoma Polygoni Cuspidati powder, add distilled water according to solid-liquid ratio 1:40,70 DEG C of preheatings are after 20 minutes, and adopt microwave auxiliary extraction, microwave power is 480W, and dutycycle is 2:3, and ultrasonic time is 20 minutes.Repeat extraction 3 times, merging filtrate.
(3) make extractum: the filtrate of extracting for 3 times merges respectively, is concentrated into fluid extract on a rotary evaporator, then in vacuum drying oven with 60 DEG C of vacuum dryings, the average quality of dry extract obtained containing Anthraquinones of Polygonum Cuspidatum Sieb is 5.04g.
(4) in extract, emodin content measures: precision takes the above-mentioned dry extract of 10mg respectively, as in 10mL volumetric flask, adds mobile phase and makees solvent and be settled to scale, shake up, filter, sample introduction 20 μ L, sample introduction 3 times, averaged.Recording emodin content in dry extract is 1.97mg/10g medical material.
Compare with comparative example from embodiments of the invention, 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 the stripping that all can promote active substance, and thus, the new method for extracting of Rhizoma Polygoni Cuspidati provided by the present invention has certain superiority.
Claims (5)
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, abundant mixing, adjust ph to 4.0 ~ 5.5, then under the stirring condition of 45 ~ 55 DEG C, enzymolysis 4 ~ 8 hours, again through high-temperature inactivation ferment treatment, finally filter;
Wherein, every 100mL hydrolytic enzyme reactant liquor to be dissolved in distilled water by cellulase according to the ratio adding 125mg cellulase in every 100mL distilled water to make;
Described high-temperature inactivation ferment treatment be by enzymolysis processing after the bath temperature of reaction system be increased to 80 DEG C, heating 10min;
2) to step 1) filter after filtering residue in add citrate-phosphate sodium dihydrogen buffer, fully mix, adjust ph to 1.5 ~ 3.0, water-bath extract after filter;
3) to step 2) filter after filtering residue in add citrate-phosphate sodium dihydrogen buffer, fully mix, adjust ph to 6.5 ~ 7.5, water-bath extract after filter;
4) to step 3) filter after filtering residue in add citrate-phosphate sodium dihydrogen buffer, fully mix, adjust ph to 8.0 ~ 8.5, water-bath extract after filter;
5) by step 1) ~ 4) filter the filtrate merging obtained, dried after recycling design, obtains the dry extract comprising Rhizoma Polygoni Cuspidati total anthraquinones;
Step 2), 3), 4) in the reaction material liquor ratio of filtering residue and citrate-phosphate sodium dihydrogen buffer be 1g:(10 ~ 22) mL.
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 2), 3), 4) described in water-bath Extracting temperature be 50 ~ 80 DEG C.
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) described in water-bath extraction time be 1 ~ 3h.
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 5) described in recycling design after dried be by merge after filtrate reduced in volume after at 60 ~ 80 DEG C, carry out vacuum drying treatment.
5. extract the method for general anthraquinone in Rhizoma Polygoni Cuspidati according to a kind of aqueous enzymatic method in Claims 1 to 4 described in any one-semi-biomimetic method, it is characterized in that, described Giant Knotweed Rhizome is after being cleaned up by Rhizoma Polygoni Cuspidati, obtained through pulverizing and sieving, drying to constant weight.
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