CN102119986A - Traditional Chinese medicine extract with anti-dementia effect and preparation method thereof - Google Patents
Traditional Chinese medicine extract with anti-dementia effect and preparation method thereof Download PDFInfo
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Abstract
The invention provides a traditional Chinese medicine extract which is prepared by ethanol extraction and macroporous absorption resin purification on Kaixinsan. The traditional Chinese medicine extract enriches saponin components and phenol components in the Kaixinsan. Meanwhile, a rat memory disorder model test proves that the traditional Chinese medicine extract provided by the invention can improve the learning and memory capacity of a model rat, so that the number of errors of the model rat is reduced and the latent period is prolonged.
Description
Technical field
The present invention relates to a kind of Chinese medicine extract, particularly a kind of Chinese medicine extract and preparation method thereof with dementia effect.
Background technology
The happy beginning of loosing sees Prescriptions Worth Thousand Gold for Emergencies, and by Radix Ginseng, Radix Polygalae each four minutes, two liang in Poria, Rhizoma Acori Graminei one or two is formed " lead and forget ".Be the representative prescription of Qi-benefiting and heart-nourishing, tranquilizing the mind, cure mainly and be traditional Chinese medical science feelings will disease: deficiency of heart-QI, mind are not peaceful, forgetful insomnia, cards such as the timid palpitation with a distress feeling of the heart.The Radix Ginseng benefit motive among this side, tranquilizing mind can be treated severe palpitation, insomnia forgetfulness; That the Radix Polygalae circulation of qi promoting is loose is strongly fragrant, Fructus Alpiniae Oxyphyllae is intelligent; All warps such as Poria GUIXIN, spleen, kidney cure mainly suffering, frightened heresy is probably throbbed with fear; The happy hole of Rhizoma Acori Graminei, improve visual and auditory acuity are dispersed the property of medicine, inducting the medicines into affected channel.At present clinically, diffusing happily and plus-minus side is used for the treatment of senile dementia and depression more.
In happy each single medicinal material that looses there be main chemical compositions: saponin (Radix Ginseng, Radix Polygalae), polysaccharide (Radix Ginseng, Poria), oligosaccharide ester (Radix Polygalae), mouthful diphenylene ketone oxide (Radix Polygalae), volatile oil (Rhizoma Acori Graminei), triterpene, (Poria, Rhizoma Acori Graminei), phenolic acid (Rhizoma Acori Graminei) etc.; At present, do not see that from happy loosing purification prepares the report of dementia active site, saponin component during the present invention will loose happily and phenols component adopt macroporous adsorbent resin to carry out enriching and purifying, obtain loosing happily the dementia active site, help the research and development of further original new drug.
Summary of the invention
The objective of the invention is to disclose a kind of Chinese medicine extract with dementia effect, another object of the present invention is open its preparation method, and the 3rd purpose of the present invention is to provide its quality determining method.
The objective of the invention is to be achieved through the following technical solutions:
Chinese medicine extract of the present invention is made by following crude drug:
Poria 200-600 weight portion Radix Ginseng 100-300 weight portion
Radix Polygalae 100-300 weight portion Rhizoma Acori Graminei 100-300 weight portion.
Chinese medicine extract of the present invention is preferably made by following crude drug:
Poria 400 weight portion Radix Ginsengs 200 weight portions
Radix Polygalae 200 weight portion Rhizoma Acori Graminei 200 weight portions.
Chinese medicine extract of the present invention is preferably made by following crude drug:
Poria 350 weight portion Radix Ginsengs 280 weight portions
Radix Polygalae 260 weight portion Rhizoma Acori Graminei 180 weight portions.
Chinese medicine extract of the present invention is preferably made by following crude drug:
Poria 560 weight portion Radix Ginsengs 160 weight portions
Radix Polygalae 180 weight portion Rhizoma Acori Graminei 270 weight portions.
The preparation method of Chinese medicine extract of the present invention comprises the steps:
Step 1: choose the aforementioned proportion crude drug;
Step 2: ethanol extraction;
Step 3: purification with macroreticular resin;
Total saponin content is 10-20% in this Chinese medicine extract, and total phenol content is 30-60%, and preferred total phenol content is 40-50%.
In the above-mentioned steps 2,, extracted 0.5-2 hour crude drug 40-80% alcohol reflux 2-4 time at every turn; Preferably use 50% alcohol reflux 3 times, each 1.5 hours;
In the above-mentioned steps 3, add aqueous dispersion dissolving with step 2 is extract obtained, making concentration of aqueous solution is 0.02-0.1g/mL, with the centrifugal 10-30min of 2000-4000 rev/min rotating speed; Preferably making concentration of aqueous solution is 0.05g/mL, with the centrifugal 10min of 3000 rev/mins rotating speed;
In the above-mentioned steps 3, get supernatant after centrifugal, by low pole or nonpolar macroporous adsorption resin, the absorption flow velocity is 1.0-3.0mL/min, and the resin column blade diameter length ratio is 0.5-1.5: 6.7-10.7, and sample solution concentration is 0.02-0.1g/mL, water elution 1-3 times of resin volume carries out remove impurity, the remove impurity flow velocity is 1.0-3.0mL/min, 3-7 times of resin volume of 70-95% ethanol elution, and elution flow rate is 1.0-3.0mL/min, collect ethanol elution, reclaim solvent, drying under reduced pressure, promptly; Preferred 80-95% ethanol elution;
In the above-mentioned steps 3, preferred 1.25L 860021 type macroporous adsorbent resins, absorption flow velocity 2.0mL/min, the resin column blade diameter length ratio is 1: 8.7, sample solution concentration is 0.05g/mL, and 2 times of resin volumes of water elution carry out remove impurity, and the remove impurity flow velocity is 1.0mL/min, 5 times of resin volumes of 90% ethanol elution, elution flow rate is 2.0mL/min, collects ethanol elution, reclaims solvent, drying under reduced pressure, promptly.
With above extract obtained, add conventional adjuvant, preparation process is routinely made the acceptable any conventional dosage form of pharmaceutics, comprises capsule, tablet, granule, gel, slow releasing agent, oral liquid.
Chinese medicine extract of the present invention is obtained through ethanol extraction, purification with macroreticular resin by happy loosing, enrichment saponin component and the phenols component in happy the loosing, pass through the evidence of rat dysmnesia model simultaneously, Chinese medicine extract of the present invention can improve the ability of learning and memory of rat model, make the rat model errors number reduce prolongation of latency.
Experimental example 1 dementia active site of the present invention causes the improvement effect of rat dysmnesia model
(1) experiment material
1. laboratory animal
The SD rat, male, body weight 256.3 ± 8.4g is provided by Beijing Vital River Experimental Animals Technology Co., Ltd., the quality certification number: SCXK (capital) 2009-2010;
2. medicine and reagent
Be subjected to reagent: the dementia active site that looses happily, press the preparation of embodiment 5 methods, dissolve wiring solution-forming with 0.5%CMC-Na before the administration.
Scopolamine, purchase company, product batch number in Fluka: 2040506, rat dosage is 0.5g/kg.
3. equipment
Rat diving tower instrument is divided into Room 2 in the case, can measure 2 rats simultaneously.
(2) experimental technique and result
1. grouping and administration
Rat is divided into 4 groups at random by body weight, be model control group, normal control group, the large and small dosage group of dementia active site of loosing happily, every group 12, the diffusing happily large and small dosage group of dementia active site successive administration 4 days, model group and normal group give isometric 0.5%CMC-Na.
2. experimental technique
(memory obtains experiment) trained in (accurately timing) lumbar injection scopolamine modeling in 30 minutes after the administration in the 4th day after 30 minutes, test after 24 hours (experiment is consolidated in memory).Test after 24 hours was trained in the modeling of model group injection scopolamine in 30 minutes.Normal group is directly trained, test after 24 hours.
Diving tower behavioristics experimentation
Will be on the same group 2 rats put into the diving tower instrument simultaneously, conformed 3 minutes, train 5 minutes, the time (incubation period) of diving tower jumped onto the first time in the monitor record, the time in Safe period, jumps off number of times (errors number) in 5 minutes.After 24 hours, remember and consolidate test, earlier rat is put on the diving tower, record jumps off the time (getting an electric shock incubation period) for the first time, the time in Safe period, jumps off number of times (errors number) in 5 minutes.
3. statistical method
Use the SAS8.0 statistical software to analyze, experimental result is represented with mean ± standard error.Adopt two sample t check and non parametric tests (Mann-Whitney Test) to compare in twos, adopt one factor analysis of variance (ANOVA) to carry out comparing between many groups, group difference adopts LSD (variance is neat) or Games-Howell method (heterogeneity of variance), to remember respectively and consolidate test phase and respectively organize administration rat and model group rat and normal rats and compare, try to achieve group difference.
4. experimental result
The not sour dementia experimental result of table 1 3-benzoyl dehydrogenation soil
Annotate: ﹠amp; ﹠amp; Compare with matched group P＜0.01; Compare with model group * P＜0.01; * compare with model group P＜0.05
(3) conclusion
Compare with model group, the large and small dosage group of dementia active site of loosing happily all can be improved the ability of learning and memory of rat model, makes the rat model errors number reduce prolongation of latency.
Following embodiment all can realize the effect of above-mentioned experimental example.
The specific embodiment
Embodiment 1: capsule
Poria 400g Radix Ginseng 200g
Radix Polygalae 200g Rhizoma Acori Graminei 200g
Get crude drug decoction pieces 1kg according to the above ratio, 50% ethanol 10L reflux, extract, 3 times was extracted 1.5 hours at every turn, and decompression and solvent recovery gets extract; Add the aqueous dispersion dissolving, making concentration of aqueous solution is that 0.05g/mL is (in folding raw medicinal herbs amount, wherein saponin concentration is 0.4009mg/ml, phenols concentration is 2.1626mg/ml), with the centrifugal 10min of 3000 rev/mins rotating speed, get supernatant, by 1.25L860021 type macroporous adsorbent resin, absorption flow velocity 2.0mL/min, the resin column blade diameter length ratio is 1: 8.7, and sample solution concentration is that (in folding raw medicinal herbs amount, wherein saponin concentration is 0.4009mg/ml to 0.05g/mL, phenols concentration is 2.1626mg/ml), 2 times of resin volumes of water elution carry out remove impurity, and the remove impurity flow velocity is 1.0mL/min, 5 times of resin volumes of 90% ethanol elution, elution flow rate is 2.0mL/min, collect 90% ethanol elution, reclaim solvent, drying under reduced pressure, add conventional adjuvant, make capsule according to common process;
The content assaying method of total saponins:
Reference substance: Radix Polygalae sapogenin;
Reagent: 0.5% vanillin-glacial acetic acid solution, perchloric acid, glacial acetic acid;
Coloration method: the accurate sample solution of drawing in right amount in cillin bottle, put behind the evaporate to dryness cold, blank retinue, after adding 0.2ml 5% vanillin-glacial acetic acid solution and 0.8ml perchloric acid successively, be incubated 15min in 60 ℃ of waters bath with thermostatic control, taking-up is put cold, after adding the 5ml glacial acetic acid, shake up, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 580 ± 2nm wave-length coverage, write down its maximum absorbance value, total saponin content is 14.94% after measured;
The total phenol content assay method:
Reference substance: 3,6 '-two mustard acyl sucrose;
Reagent: 0.6% liquor ferri trichloridi, 0.9% potassium ferricyanide solution, 0.3% sodium dodecyl sulfate solution, 70% methanol solution, 0.1mol/L hydrochloric acid solution;
Coloration method: the accurate sample solution of drawing is in right amount in the brown volumetric flask of 25ml, add 70% methanol to 5ml, add 0.3% sodium dodecyl sulfate solution 2ml again, and 1: 1 0.6% ferric chloride-0.9% potassium ferricyanide mixed solution 1ml, 5min is placed in the dark place, adds to the 25ml graduation mark with 0.1mol/L hydrochloric acid, shakes up, 30min is placed in the dark place, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 760 ± 2nm wave-length coverage, write down its maximum absorbance value, total phenol content is 45.36% after measured.
Embodiment 2: tablet
Poria 350g Radix Ginseng 280g
Radix Polygalae 260g Rhizoma Acori Graminei 180g
Get crude drug decoction pieces 1kg according to the above ratio, 60% ethanol 10L reflux, extract, 2 times was extracted 2 hours at every turn, and decompression and solvent recovery gets extract; Add the aqueous dispersion dissolving, making concentration of aqueous solution is that 0.08g/mL is (in folding raw medicinal herbs amount, wherein saponin concentration is 0.4009mg/ml, phenols concentration is 2.1626mg/ml), with the centrifugal 8min of 2500 rev/mins rotating speed, get supernatant, by 1.25L860021 type macroporous adsorbent resin, absorption flow velocity 3.0mL/min, the resin column blade diameter length ratio is 1: 6.7, and sample solution concentration is that (in folding raw medicinal herbs amount, wherein saponin concentration is 0.4009mg/ml to 0.02g/mL, phenols concentration is 2.1626mg/ml), 1.5 times of resin volumes of water elution carry out remove impurity, and the remove impurity flow velocity is 2.0mL/min, 4 times of resin volumes of 95% ethanol elution, elution flow rate is 1.0mL/min, collect ethanol elution, reclaim solvent, drying under reduced pressure, add conventional adjuvant, make tablet according to common process;
The content assaying method of total saponins:
Reference substance: Radix Polygalae sapogenin;
Reagent: 0.5% vanillin-glacial acetic acid solution, perchloric acid, glacial acetic acid;
Coloration method: the accurate sample solution of drawing in right amount in cillin bottle, put behind the evaporate to dryness cold, blank retinue, after adding 0.2ml 5% vanillin-glacial acetic acid solution and 0.8ml perchloric acid successively, be incubated 15min in 60 ℃ of waters bath with thermostatic control, taking-up is put cold, after adding the 5ml glacial acetic acid, shake up, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 580 ± 2nm wave-length coverage, write down its maximum absorbance value, total saponin content is 14.94% after measured;
The total phenol content assay method:
Reference substance: 3,6 '-two mustard acyl sucrose;
Reagent: 0.6% liquor ferri trichloridi, 0.9% potassium ferricyanide solution, 0.3% sodium dodecyl sulfate solution, 70% methanol solution, 0.1mol/L hydrochloric acid solution;
Coloration method: the accurate sample solution of drawing is in right amount in the brown volumetric flask of 25ml, add 70% methanol to 5ml, add 0.3% sodium dodecyl sulfate solution 2ml again, and 1: 1 0.6% ferric chloride-0.9% potassium ferricyanide mixed solution 1ml, 5min is placed in the dark place, adds to the 25ml graduation mark with 0.1mol/L hydrochloric acid, shakes up, 30min is placed in the dark place, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 760 ± 2nm wave-length coverage, write down its maximum absorbance value, total phenol content is 45.36% after measured.
Embodiment 3: pill
Poria 560g Radix Ginseng 160g
Radix Polygalae 180g Rhizoma Acori Graminei 270g
Get crude drug decoction pieces 1kg according to the above ratio, 70% ethanol 10L reflux, extract, 4 times was extracted 1 hour at every turn, and decompression and solvent recovery gets extract; Add the aqueous dispersion dissolving, making concentration of aqueous solution is that 0.03g/mL is (in folding raw medicinal herbs amount, wherein saponin concentration is 0.4009mg/ml, phenols concentration is 2.1626mg/ml), with the centrifugal 12min of 3800 rev/mins rotating speed, get and make supernatant, by 1.25L860021 type macroporous adsorbent resin, absorption flow velocity 1.0mL/min, the resin column blade diameter length ratio is 1: 10.7, and sample solution concentration is that (in folding raw medicinal herbs amount, wherein saponin concentration is 0.4009mg/ml to 0.1g/mL, phenols concentration is 2.1626mg/ml), 3 times of resin volumes of water elution carry out remove impurity, and the remove impurity flow velocity is 2.0mL/min, 6 times of resin volumes of 85% ethanol elution, elution flow rate is 3.0mL/min, collect ethanol elution, reclaim solvent, drying under reduced pressure, add conventional adjuvant, make pill according to common process;
The content assaying method of total saponins:
Reference substance: Radix Polygalae sapogenin;
Reagent: 0.5% vanillin-glacial acetic acid solution, perchloric acid, glacial acetic acid;
Coloration method: the accurate sample solution of drawing in right amount in cillin bottle, put behind the evaporate to dryness cold, blank retinue, after adding 0.2ml 5% vanillin-glacial acetic acid solution and 0.8ml perchloric acid successively, be incubated 15min in 60 ℃ of waters bath with thermostatic control, taking-up is put cold, after adding the 5ml glacial acetic acid, shake up, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 580 ± 2nm wave-length coverage, write down its maximum absorbance value, total saponin content is 14.94% after measured;
The total phenol content assay method:
Reference substance: 3,6 '-two mustard acyl sucrose;
Reagent: 0.6% liquor ferri trichloridi, 0.9% potassium ferricyanide solution, 0.3% sodium dodecyl sulfate solution, 70% methanol solution, 0.1mol/L hydrochloric acid solution;
Coloration method: the accurate sample solution of drawing is in right amount in the brown volumetric flask of 25ml, add 70% methanol to 5ml, add 0.3% sodium dodecyl sulfate solution 2ml again, and 1: 1 0.6% ferric chloride-0.9% potassium ferricyanide mixed solution 1ml, 5min is placed in the dark place, adds to the 25ml graduation mark with 0.1mol/L hydrochloric acid, shakes up, 30min is placed in the dark place, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 760 ± 2nm wave-length coverage, write down its maximum absorbance value, total phenol content is 45.36% after measured.
Embodiment 4: oral liquid
Poria 400g Radix Ginseng 200g
Radix Polygalae 200g Rhizoma Acori Graminei 200g
Get crude drug according to the above ratio and drink sheet 1kg, 80% ethanol 10L reflux, extract, 4 times was extracted 1 hour at every turn, and decompression and solvent recovery gets extract; Add the aqueous dispersion dissolving, making concentration of aqueous solution is that 0.1g/mL is (in folding raw medicinal herbs amount, wherein saponin concentration is 0.4009mg/ml, phenols concentration is 2.1626mg/ml), with the centrifugal 12min of 3800 rev/mins rotating speed, get supernatant, by 1.25L860021 type macroporous adsorbent resin, absorption flow velocity 2.0mL/min, the resin column blade diameter length ratio is 1: 8.7, and sample solution concentration is that (in folding raw medicinal herbs amount, wherein: saponin concentration is 0.4009mg/ml to 0.05g/mL, phenols concentration is 2.1626mg/ml), 2 times of resin volumes of water elution carry out remove impurity, and the remove impurity flow velocity is 1.0mL/min, 5 times of resin volumes of 90% ethanol elution, elution flow rate is 2.0mL/min, collect 90% ethanol elution, reclaim solvent, drying under reduced pressure, add conventional adjuvant, make oral liquid according to common process;
The content assaying method of total saponins:
Reference substance: Radix Polygalae sapogenin;
Reagent: 0.5% vanillin-glacial acetic acid solution, perchloric acid, glacial acetic acid;
Coloration method: the accurate sample solution of drawing in right amount in cillin bottle, put behind the evaporate to dryness cold, blank retinue, after adding 0.2ml 5% vanillin-glacial acetic acid solution and 0.8ml perchloric acid successively, be incubated 15min in 60 ℃ of waters bath with thermostatic control, taking-up is put cold, after adding the 5ml glacial acetic acid, shake up, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 580 ± 2nm wave-length coverage, write down its maximum absorbance value, total saponin content is 14.94% after measured;
The total phenol content assay method:
Reference substance: 3,6 '-two mustard acyl sucrose;
Reagent: 0.6% liquor ferri trichloridi, 0.9% potassium ferricyanide solution, 0.3% sodium dodecyl sulfate solution, 70% methanol solution, 0.1mol/L hydrochloric acid solution;
Coloration method: the accurate sample solution of drawing is in right amount in the brown volumetric flask of 25ml, add 70% methanol to 5ml, add 0.3% sodium dodecyl sulfate solution 2ml again, and 1: 1 0.6% ferric chloride-0.9% potassium ferricyanide mixed solution 1ml, 5min is placed in the dark place, adds to the 25ml graduation mark with 0.1mol/L hydrochloric acid, shakes up, 30min is placed in the dark place, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 760 ± 2nm wave-length coverage, write down its maximum absorbance value, total phenol content is 45.36% after measured.
Embodiment 5: injection
Poria 450g Radix Ginseng 160g
Radix Polygalae 150g Rhizoma Acori Graminei 250g
Get crude drug decoction pieces 1kg according to the above ratio, 40% ethanol 10L reflux, extract, 3 times was extracted 1.5 hours at every turn, and decompression and solvent recovery gets extract; Add the aqueous dispersion dissolving, making concentration of aqueous solution is that 0.05g/mL is (in folding raw medicinal herbs amount, wherein saponin concentration is 0.4009mg/ml, phenols concentration is 2.1626mg/ml), with the centrifugal 10min of 3000 rev/mins rotating speed, get supernatant, by 1.25L860021 type macroporous adsorbent resin, absorption flow velocity 2.0mL/min, the resin column blade diameter length ratio is 1: 8.7, and sample solution concentration is that (in folding raw medicinal herbs amount, wherein: saponin concentration is 0.4009mg/ml to 0.05g/mL, phenols concentration is 2.1626mg/ml), 2 times of resin volumes of water elution carry out remove impurity, and the remove impurity flow velocity is 1.0mL/min, 5 times of resin volumes of 90% ethanol elution, elution flow rate is 2.0mL/min, collect 90% ethanol elution, reclaim solvent, drying under reduced pressure, add conventional adjuvant, make injection according to common process;
The content assaying method of total saponins:
Reference substance: Radix Polygalae sapogenin;
Reagent: 0.5% vanillin-glacial acetic acid solution, perchloric acid, glacial acetic acid;
Coloration method: the accurate sample solution of drawing in right amount in cillin bottle, put behind the evaporate to dryness cold, blank retinue, after adding 0.2ml 5% vanillin-glacial acetic acid solution and 0.8ml perchloric acid successively, be incubated 15min in 60 ℃ of waters bath with thermostatic control, taking-up is put cold, after adding the 5ml glacial acetic acid, shake up, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 580 ± 2nm wave-length coverage, write down its maximum absorbance value, total saponin content is 14.94% after measured;
The total phenol content assay method:
Reference substance: 3,6 '-two mustard acyl sucrose;
Reagent: 0.6% liquor ferri trichloridi, 0.9% potassium ferricyanide solution, 0.3% sodium dodecyl sulfate solution, 70% methanol solution, 0.1mol/L hydrochloric acid solution;
Coloration method: the accurate sample solution of drawing is in right amount in the brown volumetric flask of 25ml, add 70% methanol to 5ml, add 0.3% sodium dodecyl sulfate solution 2ml again, and 1: 1 0.6% ferric chloride-0.9% potassium ferricyanide mixed solution 1ml, 5min is placed in the dark place, adds to the 25ml graduation mark with 0.1mol/L hydrochloric acid, shakes up, 30min is placed in the dark place, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 760 ± 2nm wave-length coverage, write down its maximum absorbance value, total phenol content is 45.36% after measured.
Embodiment 6:
Poria 400g Radix Ginseng 200g
Radix Polygalae 200g Rhizoma Acori Graminei 200g
Get crude drug decoction pieces 1kg according to the above ratio, 50% ethanol 10L reflux, extract, 3 times was extracted 1.5 hours at every turn, and decompression and solvent recovery gets extract; Add the aqueous dispersion dissolving, making concentration of aqueous solution is that 0.05g/mL is (in folding raw medicinal herbs amount, wherein saponin concentration is 0.4009mg/ml, phenols concentration is 2.1626mg/ml), with the centrifugal 10min of 3000 rev/mins rotating speed, get supernatant, by 1.25L860021 type macroporous adsorbent resin, absorption flow velocity 2.0mL/min, the resin column blade diameter length ratio is 1: 8.7, sample solution concentration is 0.05g/mL (in folding raw medicinal herbs amount, wherein saponin concentration is 0.4009mg/ml, and phenols concentration is 2.1626mg/ml), 2 times of resin volumes of water elution carry out remove impurity, the remove impurity flow velocity is 1.0mL/min, 5 times of resin volumes of 90% ethanol elution, and elution flow rate is 2.0mL/min, collect 90% ethanol elution, reclaim solvent, drying under reduced pressure is Chinese medicine extract of the present invention;
The content assaying method of total saponins:
Reference substance: Radix Polygalae sapogenin;
Reagent: 0.5% vanillin-glacial acetic acid solution, perchloric acid, glacial acetic acid;
Coloration method: the accurate sample solution of drawing in right amount in cillin bottle, put behind the evaporate to dryness cold, blank retinue, after adding 0.2ml 5% vanillin-glacial acetic acid solution and 0.8ml perchloric acid successively, be incubated 15min in 60 ℃ of waters bath with thermostatic control, taking-up is put cold, after adding the 5ml glacial acetic acid, shake up, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 580 ± 2nm wave-length coverage, write down its maximum absorbance value, total saponin content is 14.94% after measured;
The total phenol content assay method:
Reference substance: 3,6 '-two mustard acyl sucrose;
Reagent: 0.6% liquor ferri trichloridi, 0.9% potassium ferricyanide solution, 0.3% sodium dodecyl sulfate solution, 70% methanol solution, 0.1mol/L hydrochloric acid solution;
Coloration method: the accurate sample solution of drawing is in right amount in the brown volumetric flask of 25ml, add 70% methanol to 5ml, add 0.3% sodium dodecyl sulfate solution 2ml again, and 1: 1 0.6% ferric chloride-0.9% potassium ferricyanide mixed solution 1ml, 5min is placed in the dark place, adds to the 25ml graduation mark with 0.1mol/L hydrochloric acid, shakes up, 30min is placed in the dark place, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 760 ± 2nm wave-length coverage, write down its maximum absorbance value, total phenol content is 45.36% after measured.
Embodiment 7:
Poria 350g Radix Ginseng 280g
Radix Polygalae 260g Rhizoma Acori Graminei 180g
Get crude drug decoction pieces 1kg according to the above ratio, 60% ethanol 10L reflux, extract, 2 times was extracted 2 hours at every turn, and decompression and solvent recovery gets extract; Add the aqueous dispersion dissolving, making concentration of aqueous solution is that 0.08g/mL is (in folding raw medicinal herbs amount, wherein saponin concentration is 0.4009mg/ml, phenols concentration is 2.1626mg/ml), with the centrifugal 8min of 2500 rev/mins rotating speed, get supernatant, by 1.25L860021 type macroporous adsorbent resin, absorption flow velocity 3.0mL/min, the resin column blade diameter length ratio is 1: 6.7, sample solution concentration is 0.02g/mL (in folding raw medicinal herbs amount, wherein saponin concentration is 0.4009mg/ml, and phenols concentration is 2.1626mg/ml), 1.5 times of resin volumes of water elution carry out remove impurity, the remove impurity flow velocity is 2.0mL/min, 4 times of resin volumes of 95% ethanol elution, and elution flow rate is 1.0mL/min, collect ethanol elution, reclaim solvent, drying under reduced pressure is Chinese medicine extract of the present invention;
The content assaying method of total saponins:
Reference substance: Radix Polygalae sapogenin;
Reagent: 0.5% vanillin-glacial acetic acid solution, perchloric acid, glacial acetic acid;
Coloration method: the accurate sample solution of drawing in right amount in cillin bottle, put behind the evaporate to dryness cold, blank retinue, after adding 0.2ml 5% vanillin-glacial acetic acid solution and 0.8ml perchloric acid successively, be incubated 15min in 60 ℃ of waters bath with thermostatic control, taking-up is put cold, after adding the 5ml glacial acetic acid, shake up, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 580 ± 2nm wave-length coverage, write down its maximum absorbance value, total saponin content is 14.94% after measured;
The total phenol content assay method:
Reference substance: 3,6 '-two mustard acyl sucrose;
Reagent: 0.6% liquor ferri trichloridi, 0.9% potassium ferricyanide solution, 0.3% sodium dodecyl sulfate solution, 70% methanol solution, 0.1mol/L hydrochloric acid solution;
Coloration method: the accurate sample solution of drawing is in right amount in the brown volumetric flask of 25ml, add 70% methanol to 5ml, add 0.3% sodium dodecyl sulfate solution 2ml again, and 1: 1 0.6% ferric chloride-0.9% potassium ferricyanide mixed solution 1ml, 5min is placed in the dark place, adds to the 25ml graduation mark with 0.1mol/L hydrochloric acid, shakes up, 30min is placed in the dark place, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 760 ± 2nm wave-length coverage, write down its maximum absorbance value, total phenol content is 45.36% after measured.
Embodiment 8:
Poria 560g Radix Ginseng 160g
Radix Polygalae 180g Rhizoma Acori Graminei 270g
Get crude drug decoction pieces 1kg according to the above ratio, 70% ethanol 10L reflux, extract, 4 times was extracted 1 hour at every turn, and decompression and solvent recovery gets extract; Add the aqueous dispersion dissolving, making concentration of aqueous solution is that 0.03g/mL is (in folding raw medicinal herbs amount, wherein saponin concentration is 0.4009mg/ml, phenols concentration is 2.1626mg/ml), with the centrifugal 12min of 3800 rev/mins rotating speed, get and make supernatant, by 1.25L860021 type macroporous adsorbent resin, absorption flow velocity 1.0mL/min, the resin column blade diameter length ratio is 1: 10.7, sample solution concentration is 0.1g/mL (in folding raw medicinal herbs amount, wherein saponin concentration is 0.4009mg/ml, and phenols concentration is 2.1626mg/ml), 3 times of resin volumes of water elution carry out remove impurity, the remove impurity flow velocity is 2.0mL/min, 6 times of resin volumes of 85% ethanol elution, and elution flow rate is 3.0mL/min, collect ethanol elution, reclaim solvent, drying under reduced pressure is Chinese medicine extract of the present invention;
The content assaying method of total saponins:
Reference substance: Radix Polygalae sapogenin;
Reagent: 0.5% vanillin-glacial acetic acid solution, perchloric acid, glacial acetic acid;
Coloration method: the accurate sample solution of drawing in right amount in cillin bottle, put behind the evaporate to dryness cold, blank retinue, after adding 0.2ml 5% vanillin-glacial acetic acid solution and 0.8ml perchloric acid successively, be incubated 15min in 60 ℃ of waters bath with thermostatic control, taking-up is put cold, after adding the 5ml glacial acetic acid, shake up, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 580 ± 2nm wave-length coverage, write down its maximum absorbance value, total saponin content is 14.94% after measured;
The total phenol content assay method:
Reference substance: 3,6 '-two mustard acyl sucrose;
Reagent: 0.6% liquor ferri trichloridi, 0.9% potassium ferricyanide solution, 0.3% sodium dodecyl sulfate solution, 70% methanol solution, 0.1mol/L hydrochloric acid solution;
Coloration method: the accurate sample solution of drawing is in right amount in the brown volumetric flask of 25ml, add 70% methanol to 5ml, add 0.3% sodium dodecyl sulfate solution 2ml again, and 1: 1 0.6% ferric chloride-0.9% potassium ferricyanide mixed solution 1ml, 5min is placed in the dark place, adds to the 25ml graduation mark with 0.1mol/L hydrochloric acid, shakes up, 30min is placed in the dark place, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 760 ± 2nm wave-length coverage, write down its maximum absorbance value, total phenol content is 45.36% after measured.
Embodiment 9:
Poria 400g Radix Ginseng 200g
Radix Polygalae 200g Rhizoma Acori Graminei 200g
Get crude drug according to the above ratio and drink sheet 1kg, 80% ethanol 10L reflux, extract, 4 times was extracted 1 hour at every turn, and decompression and solvent recovery gets extract; Add the aqueous dispersion dissolving, making concentration of aqueous solution is that 0.1g/mL is (in folding raw medicinal herbs amount, wherein saponin concentration is 0.4009mg/ml, phenols concentration is 2.1626mg/ml), with the centrifugal 12min of 3800 rev/mins rotating speed, get supernatant, by 1.25L860021 type macroporous adsorbent resin, absorption flow velocity 2.0mL/min, the resin column blade diameter length ratio is 1: 8.7, sample solution concentration is 0.05g/mL (in folding raw medicinal herbs amount, wherein: saponin concentration is 0.4009mg/ml, and phenols concentration is 2.1626mg/ml), 2 times of resin volumes of water elution carry out remove impurity, the remove impurity flow velocity is 1.0mL/min, 5 times of resin volumes of 90% ethanol elution, and elution flow rate is 2.0mL/min, collect 90% ethanol elution, reclaim solvent, drying under reduced pressure is Chinese medicine extract of the present invention;
The content assaying method of total saponins:
Reference substance: Radix Polygalae sapogenin;
Reagent: 0.5% vanillin-glacial acetic acid solution, perchloric acid, glacial acetic acid;
Coloration method: the accurate sample solution of drawing in right amount in cillin bottle, put behind the evaporate to dryness cold, blank retinue, after adding 0.2ml 5% vanillin-glacial acetic acid solution and 0.8ml perchloric acid successively, be incubated 15min in 60 ℃ of waters bath with thermostatic control, taking-up is put cold, after adding the 5ml glacial acetic acid, shake up, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 580 ± 2nm wave-length coverage, write down its maximum absorbance value, total saponin content is 14.94% after measured;
The total phenol content assay method:
Reference substance: 3,6 '-two mustard acyl sucrose;
Reagent: 0.6% liquor ferri trichloridi, 0.9% potassium ferricyanide solution, 0.3% sodium dodecyl sulfate solution, 70% methanol solution, 0.1mol/L hydrochloric acid solution;
Coloration method: the accurate sample solution of drawing is in right amount in the brown volumetric flask of 25ml, add 70% methanol to 5ml, add 0.3% sodium dodecyl sulfate solution 2ml again, and 1: 1 0.6% ferric chloride-0.9% potassium ferricyanide mixed solution 1ml, 5min is placed in the dark place, adds to the 25ml graduation mark with 0.1mol/L hydrochloric acid, shakes up, 30min is placed in the dark place, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 760 ± 2nm wave-length coverage, write down its maximum absorbance value, total phenol content is 45.36% after measured.
Embodiment 10:
Poria 450g Radix Ginseng 160g
Radix Polygalae 150g Rhizoma Acori Graminei 250g
Get crude drug decoction pieces 1kg according to the above ratio, 40% ethanol 10L reflux, extract, 3 times was extracted 1.5 hours at every turn, and decompression and solvent recovery gets extract; Add the aqueous dispersion dissolving, making concentration of aqueous solution is that 0.05g/mL is (in folding raw medicinal herbs amount, wherein saponin concentration is 0.4009mg/ml, phenols concentration is 2.1626mg/ml), with the centrifugal 10min of 3000 rev/mins rotating speed, get supernatant, by 1.25L860021 type macroporous adsorbent resin, absorption flow velocity 2.0mL/min, the resin column blade diameter length ratio is 1: 8.7, sample solution concentration is 0.05g/mL (in folding raw medicinal herbs amount, wherein: saponin concentration is 0.4009mg/ml, and phenols concentration is 2.1626mg/ml), 2 times of resin volumes of water elution carry out remove impurity, the remove impurity flow velocity is 1.0mL/min, 5 times of resin volumes of 90% ethanol elution, and elution flow rate is 2.0mL/min, collect 90% ethanol elution, reclaim solvent, drying under reduced pressure is Chinese medicine extract of the present invention;
The content assaying method of total saponins:
Reference substance: Radix Polygalae sapogenin;
Reagent: 0.5% vanillin-glacial acetic acid solution, perchloric acid, glacial acetic acid;
Coloration method: the accurate sample solution of drawing in right amount in cillin bottle, put behind the evaporate to dryness cold, blank retinue, after adding 0.2ml 5% vanillin-glacial acetic acid solution and 0.8ml perchloric acid successively, be incubated 15min in 60 ℃ of waters bath with thermostatic control, taking-up is put cold, after adding the 5ml glacial acetic acid, shake up, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 580 ± 2nm wave-length coverage, write down its maximum absorbance value, total saponin content is 14.94% after measured;
The total phenol content assay method:
Reference substance: 3,6 '-two mustard acyl sucrose;
Reagent: 0.6% liquor ferri trichloridi, 0.9% potassium ferricyanide solution, 0.3% sodium dodecyl sulfate solution, 70% methanol solution, 0.1mol/L hydrochloric acid solution;
Coloration method: the accurate sample solution of drawing is in right amount in the brown volumetric flask of 25ml, add 70% methanol to 5ml, add 0.3% sodium dodecyl sulfate solution 2ml again, and 1: 1 0.6% ferric chloride-0.9% potassium ferricyanide mixed solution 1ml, 5min is placed in the dark place, adds to the 25ml graduation mark with 0.1mol/L hydrochloric acid, shakes up, 30min is placed in the dark place, in order to measuring;
Assay method: sample after the above-mentioned colour developing is measured in 760 ± 2nm wave-length coverage, write down its maximum absorbance value, total phenol content is 45.36% after measured.
Claims (10)
1. Chinese medicine extract with dementia effect is characterized in that mainly being made by following method of this Chinese medicine extract:
Step 1: choose following crude drug:
Poria 200-600 weight portion Radix Ginseng 100-300 weight portion
Radix Polygalae 100-300 weight portion Rhizoma Acori Graminei 100-300 weight portion;
Step 2: ethanol extraction;
Step 3: low pole or nonpolar macroporous adsorption resin purification;
Wherein total saponin content is 10-20%, and total phenol content is 30-60%.
2. Chinese medicine extract as claimed in claim 1 is characterized in that consisting of of crude drug in the step 1:
3. Chinese medicine extract as claimed in claim 1 or 2 is characterized in that in the step 2, with crude drug 40-80% alcohol reflux 2-4 time, extracts 0.5-2 hour at every turn.
4. Chinese medicine extract as claimed in claim 1 or 2 is characterized in that in the step 3, adds aqueous dispersion dissolving with step 2 is extract obtained, and making concentration of aqueous solution is 0.02-0.1g/mL, with the centrifugal 10-30min of 2000-4000 rev/min rotating speed.
5. Chinese medicine extract as claimed in claim 1 or 2, it is characterized in that in the step 3, get supernatant after centrifugal, by low pole or nonpolar macroporous adsorption resin, the absorption flow velocity is 1.0-3.0mL/min, the resin column blade diameter length ratio is 0.5-1.5: 6.7-10.7, sample solution concentration is 0.02-0.1g/mL, and water elution 1-3 times of resin volume carries out remove impurity, and the remove impurity flow velocity is 1.0-3.0mL/min, 3-7 times of resin volume of 70-95% ethanol elution, elution flow rate is 1.0-3.0mL/min, collects ethanol elution, reclaims solvent, drying under reduced pressure, promptly.
6. the preparation method with Chinese medicine extract of dementia effect is characterized in that this method comprises the steps:
Step 1: choose following crude drug:
Poria 200-600 weight portion Radix Ginseng 100-300 weight portion
Radix Polygalae 100-300 weight portion Rhizoma Acori Graminei 100-300 weight portion;
Step 2: ethanol extraction;
Step 3: low pole or nonpolar macroporous adsorption resin purification.
7. method as claimed in claim 6 is characterized in that in the step 2, with crude drug 40-80% alcohol reflux 2-4 time, extracts 0.5-2 hour at every turn.
8. method as claimed in claim 6 is characterized in that in the step 3, adds aqueous dispersion dissolving with step 2 is extract obtained, and making concentration of aqueous solution is 0.02-0.1g/mL, with the centrifugal 10-30min of 2000-4000 rev/min rotating speed.
9. method as claimed in claim 6, it is characterized in that in the step 3, get supernatant after centrifugal, by low pole or nonpolar macroporous adsorption resin, the absorption flow velocity is 1.0-3.0mL/min, the resin column blade diameter length ratio is 0.5-1.5: 6.7-10.7, sample solution concentration is 0.02-0.1g/mL, and water elution 1-3 times of resin volume carries out remove impurity, and the remove impurity flow velocity is 1.0-3.0mL/min, 3-7 times of resin volume of 70-95% ethanol elution, elution flow rate is 1.0-3.0mL/min, collects ethanol elution, reclaims solvent, drying under reduced pressure, promptly.
10. has application in the dementia drugs with function as the arbitrary described Chinese medicine extract of claim 1-5 in preparation.
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| CN103977163A (en) * | 2014-05-27 | 2014-08-13 | 石任兵 | Pharmaceutical composition with myocardial ischemia resisting effect and preparation method and application thereof |
| CN105687564A (en) * | 2016-02-23 | 2016-06-22 | 广西梧州制药（集团）股份有限公司 | Traditional Chinese medicine composition for improving sleep and memory and preparation method of traditional Chinese medicine composition for improving sleep and memory |
| CN107184776A (en) * | 2017-06-07 | 2017-09-22 | 吉林天药本草堂制药有限公司 | The black brain tonic pharmaceutical composition of ginseng |
| CN110623969A (en) * | 2019-09-05 | 2019-12-31 | 河北中医学院 | Component formula for treating amnesia |
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| CN103977179B (en) * | 2014-05-27 | 2021-10-01 | 石任兵 | Medicine substance with anti-dementia effect and preparation method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103099911A (en) * | 2012-12-28 | 2013-05-15 | 广东万年青制药有限公司 | Application of mental treasure pills in preparation of medicine for treating senile dementia |
| CN103977163A (en) * | 2014-05-27 | 2014-08-13 | 石任兵 | Pharmaceutical composition with myocardial ischemia resisting effect and preparation method and application thereof |
| CN103977163B (en) * | 2014-05-27 | 2021-04-20 | 石任兵 | Medicinal composition with myocardial ischemia resisting effect, preparation method and application thereof |
| CN105687564A (en) * | 2016-02-23 | 2016-06-22 | 广西梧州制药（集团）股份有限公司 | Traditional Chinese medicine composition for improving sleep and memory and preparation method of traditional Chinese medicine composition for improving sleep and memory |
| CN107184776A (en) * | 2017-06-07 | 2017-09-22 | 吉林天药本草堂制药有限公司 | The black brain tonic pharmaceutical composition of ginseng |
| CN110623969A (en) * | 2019-09-05 | 2019-12-31 | 河北中医学院 | Component formula for treating amnesia |
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