CN100560083C - A kind of preparation method of from the Chinese medicine Radix Notoginseng, extracting purified general saponin - Google Patents
A kind of preparation method of from the Chinese medicine Radix Notoginseng, extracting purified general saponin Download PDFInfo
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Abstract
A kind of preparation method of extracting purified general saponin from the Chinese medicine Radix Notoginseng by pseudo-ginseng being ground into powder or granule, is removed impurity, oven dry, with containing the water-soluble matchmaker's diafiltration of ethanol that concentration is 0-95%, hot reflux is extracted, the enrichment of extracting solution concentrating under reduced pressure is with D on the concentrated solution
101Macroporous resin successively with water, ammonia, pure water elution, is collected eluent, concentrates, and drying promptly gets the refining thing of Radix Notoginseng total arasaponins.The Radix Notoginseng total arasaponins that method of the present invention is made can be made into various medicaments such as tablet, capsule, injection and goes up said dosage form, also can cooperate other medicines or component to make preparation together and use.The inventive method only needs ethanol water to extract, and uses a macroporous resin column to separate, end-product Radix Notoginseng total arasaponins purity height, and product stability and homogeneity are good.The inventive method is reasonable in design, and low-cost, easy to operate, good stability, Quality Control be stricter, be easy to industrialization production.
Description
Technical field
The invention belongs to field of medicaments, particularly relate to from the Chinese medicine Radix Notoginseng through extracting, make with extra care, the acquisition Radix Notoginseng total arasaponins.
Background technology
At present, in the world, Chinese herbal medicine all has vast market, along with people increasing and the aging of population to the health requirements level of understanding, sub-health stateization, people thirst for back to nature more, utilize the natural plants treatment, prevent the insoluble disease of some chemical synthetic drugs.From natural drug, seek the little and inexpensive medicine of side effect and become the target that countries in the world pharmaceutical manufacturer is chased.The European Community has carried out unified legislation to medical herbs, state medical herbs status such as Canada and Australia have legalized, U.S. government has also drafted the plant amedica management method, the compound recipe mix preparation that begins to accept natural drug is as curative, and these provide good international environment for Chinese medicine enters international medical market as medicine.On the other hand, along with the quickening of global economic integration progress, particularly China becomes a full member of WTO, and the breadth and depth that Chinese Medicine market incorporates international medical big market will further strengthen.Face the enormous impact of Asian countries's traditional medicine product such as the keen competition of powerful transnational medical group and Japan, Korea S, India, Thailand and European countries' plant amedica such as Germany, France, numerous products that China's Chinese medicine produces are owing to still can not meet the standard of international medical market and requirement and being kept outside of the door.
Radix Notoginseng is the dry root of panax araliaceae plant Panax notoginseng (Burk.) F.H.Chen.The traditional Chinese medical science is thought, its sweet in the mouth, little hardship are warm in nature, return liver, stomach warp.Have the dissipating blood stasis hemostasis, the effect of subduing swelling and relieving pain is applicable to the treatment spitting of blood, spits blood, and epistaxis is had blood in stool, metrorrhagia, traumatic hemorrhage, breast ventral spine pain, tumbling and swelling etc.Modern study shows, mainly contain ginsenoside (Ginsenosides) Rb1, Rd, Re, Rg1, Rg2, Rh in the Radix Notoginseng, 20-O-glucose ginsenoside Rf, arasaponin (Notoginsenoside) R1, R2, R3, R4, Herba Gynostemmatis saponin (Gypeno-side) X VII still contains the ginsenoside Rb2.Also contain volatile oil, have α-and β-guaiene (α-, β-guaiene), α-copaene (α-copaene) is arranged in addition, δ-guaiene, chamigrene (cuparene), α-, β-, γ-elemene (α-, β-, γ-elemene), α-Cananga odorata oil alkene (α-muurolene), α-Gu Yunxi (α-gurjunene), methyl palmitate, ethyl palmitate, the heptadecadienoic acid methyl ester, octadecadienoic ethyl ester, phthalic acid di tert butyl carbonate (benzene-diformic acid ditert-butylate), octadecadienoic acid, isopropylbenzene, encircle 12 carbon ketone (cy-clodode-canone), 1-methyl-4-peroxide methyl mercapto [2,2,2] octane, the tetradecane, hexadecane, heptadecane, octadecane, nonadecane, larane, 21 alkane, docosane etc.The dencichine (N-oxalo-L-α, β-diaminopro-pionic acid) that contains a kind of tool styptic activity in the aqueous extract.Other contains Quercetin, Radix Notoginseng flavone B, cupreol, cupreol-D-glucoside, sucrose.Saponin component in the Radix Notoginseng is that research is maximum at present, and the most significant composition of drug activity has heart tonifying, and vasoconstrictive shortens the effect of clotting time and thrombinogen formation time, now has been used for the treatment of purpura hemorrhagica.Influence to blood pressure is to raise earlier afterwards to make it to descend.Heavy dose of energy blood vessel dilating, coronary blood flow increasing, decreased heart rate reduces myocardial oxygen consumption, now has been used for the treatment of coronary heart disease.
At present, the technology of preparation total saponins still has some documents and patent report from Radix Notoginseng, but in general, exists processing step loaded down with trivial details, and product purity is not high, the low three aspect deficiencies of quality control level.
Prior art prepares the step of total saponins from Radix Notoginseng comparatively loaded down with trivial details or the committed step specification requirement is higher, is difficult to realize that suitability for industrialized production or production cost are too high.As the disclosed method of patent 1 (CN01108591.6) in order to remove the materials such as saccharide, flavonoid and pigment in the Radix Notoginseng, used the resin of two types in macroporous resin and ion exchange resin, complex process is through multiple tracks steps such as extraction, clarification, decolouring, desaccharide, spray dryinges; Patent 2 (CN93105774.4) has adopted immersion, pulverizing, lixiviate, fine straining especially, has concentrated, chromatography, decolouring, filtration, concentrate, ten procedures such as drying, and successively used aluminium oxide and two kinds of depigmenting agents of active carbon, drying has then been used acetone drying and vacuum drying; The disclosed method of patent 3 (CN02131193.5) has used quaternary amine base resin to carry out purification.Various product loss increase, the yield of not only can causing of processing step descends, and also can have influence on industrial production efficiency, improves production cost.The disclosed method of patent 4 (CN200410091532.7) has then used the preparative high performance liquid chromatography instrument to separate.Working condition present stage that preparation liquid phase etc. is had relatively high expectations can not promote the use in commercial production.The method disclosed in the present is easy, and is workable, only needs to extract after purification by macroporous resin gets final product, and is beneficial to the large-scale production utilization, and can significantly reduces production costs.
Problems such as in addition, existing Technology prepares Radix Notoginseng total arasaponins and exists purity not high, and target level of product quality is low.What the assay of total saponins adopted in the regulation Radix Notoginseng in the Pharmacopoeia of People's Republic of China (2005 editions first one) is high performance liquid chromatography, with respect to traditional ultraviolet spectrophotometry, its accuracy and specificity are all excellent, and patent 1 (CN01108591.6) adopts traditional ultraviolet spectrophotometry to come the total saponins quality is estimated, and only with high-efficient liquid phase technique to Rg wherein
1, Rb
1, R
1Three monomer saponins carry out assay.Saponin content is respectively 88% and 60% in the middle use of patent 2 (CN93105774.4) and patent 3 (CN02131193.5) the high effective liquid chromatography for measuring product, controlled component content is not high, this must influence Radix Notoginseng total arasaponins at clinical application safety and effectiveness, also is unfavorable for breaking through the technology barriers that enter the international market.The method disclosed in the present has realized high-purity, high yield on the basis of simple procedures, use high-efficient liquid phase technique to measure total saponins, and purity can reach 95%, and the rate of transform of total saponins in from the medical material to the product is more than 90%.Especially in the purification by macroporous resin process, adopt certain density ammonia spirit flushing on the sample behind the sample through a large amount of experiment showed,, can effectively remove phenolic constituent, greatly improved the content (the experiment proved that total saponin content can bring up to 95% from 80%) of total saponins.
Summary of the invention
The objective of the invention is to overcome the deficiency that prior art exists, a kind of preparation method of extracting purified general saponin from the Chinese medicine Radix Notoginseng is provided, comprise that medical material extracts with alcohol-water solution, extracting solution concentrates uses the macroporous adsorptive resins separation steps.
Specifically be achieved through the following technical solutions:
(1) pseudo-ginseng was ground into the powder of 20 mesh sieves or particle diameter less than the granule of 1cm, removed impurity, and oven dry is with containing the water-soluble matchmaker's diafiltration of ethanol that concentration is 0-95%, ultrasonic, hot reflux is extracted, and solvent load 5-20 doubly measures, extract 1-3 time, each 0.5-2 hour, the enrichment of extracting solution concentrating under reduced pressure;
(2) D on the concentrated solution
101Macroporous resin, successively with water, ammonia, pure water elution, first water flushing 3-6 column volume (BV), reuse 0.05%-3% ammonia flushing 3-6BV, reuse water washes to pH=7, uses 20-80% ethanol elution 4-8BV at last, collects eluent, concentrate, drying promptly gets the refining thing of Radix Notoginseng total arasaponins.
Pseudo-ginseng is ground into the graininess of particle diameter 1mm-10mm in the step (1), and to contain the water-soluble matchmaker's diafiltration of ethanol that concentration is 30-95%, solvent load 6-15 doubly measures, and hot reflux is extracted 1-2 time, each 1-2 hour.
In the step (1), pseudo-ginseng is ground into the graininess of 3mm-8mm, and to contain the ethanol water that concentration is 65-75%, solvent load 10-12 doubly measures, and hot reflux is extracted 2 times, each 2 hours.
D on the sample in the step (2)
101Macroporous resin, 4-6 column volume of elder generation's water flushing, 4-6 column volume of reuse 0.05%-1% ammonia flushing, reuse water washes to pH=7,3-5 column volume of reuse 20-30% alcohol flushing, use 3-6 column volume of 60-80% ethanol elution at last, collect eluent, reclaim solvent and get the refining thing of arasaponin.
In the step (2), on the sample behind the sample, elder generation's water flushing 4-5 column volume (BV), reuse 0.05-0.1% ammonia flushing 4-5BV, reuse water washes to pH=7, reuse 20-30% alcohol flushing 4-5BV, use 60-70% ethanol elution 5-6BV at last, collect eluent, concentrate, drying gets the refining thing of Radix Notoginseng total arasaponins.
According to the Radix Notoginseng total arasaponins that method of the present invention is made, can be made into various medicaments such as tablet, capsule, injection and go up said dosage form, also can cooperate other medicines or component to make preparation together and use.
The usefulness of the method for the invention is: described method is easy and simple to handle, and extracting only needs ethanol water, and separating only needs to use a macroporous resin column; End-product Radix Notoginseng total arasaponins purity height can reach more than 95%; When crossing macroporous resin column, with the ammonia flushing, can effectively remove flavone in the Radix Notoginseng, liposoluble ingredient, improve saponin yield (the total saponins yield is brought up to more than 95% from 80%); To 5 main component R in the total saponins
1, Rg
1, Re, Rd, Rb
1Carry out assay with HPLC, simultaneously HPLC fingerprint map analyzing result proves that product stability and homogeneity are good.The inventive method is reasonable in design, and low-cost, easy to operate, good stability, Quality Control be stricter, be easy to industrialization production.
Description of drawings
Fig. 1 is that the highest monomer saponin of content carries out quantitative analysis in five kinds of Radix Notoginseng.
The specific embodiment
The present invention is further described in conjunction with the embodiments, should be appreciated that following each embodiment only is used to the present invention is described and is not limitation of the present invention.
Embodiment one
Get pseudo-ginseng coarse granule (the about 1cm of particle diameter) 500g, 50 ℃ of dryings 6 hours add 8 times of amount 30% ethanol slight boiling condition heating and refluxing extraction 2 times, and each 1 hour, extracting liquid filtering merged, and is evaporated to 0.5g/mL (crude drug amount), last D
101Macroporous resin.With 4 times of amount column volume deionized water rinsings, discard, continue to discard with the flushing of 4BV 30% alcoholic solution, use 5BV 70% alcoholic solution eluting at last, collect eluent, being evaporated to does not have alcohol, and lyophilization promptly gets Radix Notoginseng total arasaponins.Measure Panax Notoginseng saponin R in the refining thing through HPLC
12%, ginsenoside Rg
115%, ginsenoside Rb
19%, ginsenoside Rd 3%, ginsenoside Re 1%, total saponin content reaches 30%.
Embodiment two
Get pseudo-ginseng coarse granule (the about 1cm of particle diameter) 500g, 50 ℃ of dryings 6 hours add 15 times of amount 40% ethanol slight boiling condition heating and refluxing extraction 1 time, and 2 hours, extracting liquid filtering merged, and is evaporated to 0.5g/mL (crude drug amount), last D
101Macroporous resin.With 5 times of amount column volume deionized water rinsings, discard, the flushing of reuse 4BV 1% ammonia discards, then with deionized water rinsing to effluent pH=7, use 6BV 30% alcoholic solution eluting at last, collect eluent, being evaporated to does not have alcohol, and lyophilization promptly gets Radix Notoginseng total arasaponins.Measure Panax Notoginseng saponin R in the refining thing through HPLC
14%, ginsenoside Rg
130%, ginsenoside Rb
115%, ginsenoside Rd 5%, ginsenoside Re 3%, total saponin content reaches 57%.
Embodiment three
Get pseudo-ginseng coarse granule (the about 1cm of particle diameter) 500g, 50 ℃ of dryings 6 hours add 18 times of amount 50% ethanol slight boiling condition heating and refluxing extraction 1 time, and 1 hour, extracting liquid filtering merged, and is evaporated to 0.5g/mL (crude drug amount), last D
101Macroporous resin.With 4 times of amount column volume deionized water rinsings, discard, the flushing of reuse 4BV 0.1% ammonia discards, then with deionized water rinsing to effluent pH=7, use 5BV 60% alcoholic solution eluting at last, collect eluent, being evaporated to does not have alcohol, and lyophilization promptly gets Radix Notoginseng total arasaponins.Measure Panax Notoginseng saponin R in the refining thing through HPLC
14%, ginsenoside Rg
126%, ginsenoside Rb
116%, ginsenoside Rd 5%, ginsenoside Re 4%, total saponin content reaches 55%.
Embodiment four
Get pseudo-ginseng granule (1mm-10mm) 500g, 50 ℃ of dryings 6 hours add 12 times of amount 50% ethanol slight boiling condition heating and refluxing extraction 2 times, and each 2 hours, extracting liquid filtering merged, and is evaporated to 0.5g/mL (crude drug amount), last D
101Macroporous resin.With 3 times of amount column volume deionized water rinsings, discard, the flushing of reuse 3BV 0.05% ammonia discards, then with deionized water rinsing to effluent pH=7, continue to discard, use 5BV 50% alcoholic solution eluting at last with the flushing of 4BV 20% alcoholic solution, collect eluent, being evaporated to does not have alcohol, and lyophilization promptly gets Radix Notoginseng total arasaponins.Through the HPLC external standard method, Panax Notoginseng saponin R in the refining thing
16%, ginsenoside Rg
142%, ginsenoside Rb
123%, ginsenoside Rd 7%, ginsenoside Re 3%, total saponin content reaches 81%.
Embodiment five
Get pseudo-ginseng granule (1mm-10mm) 500g, 50 ℃ of dryings 6 hours add 15 times of amount 60% ethanol slight boiling condition heating and refluxing extraction 1 time, and 1 hour, extracting liquid filtering merged, and is evaporated to 0.5g/mL (crude drug amount), last D
101Macroporous resin.With 3 times of amount column volume deionized water rinsings, discard, the flushing of reuse 5BV 0.5% ammonia discards, then with deionized water rinsing to effluent pH=7, continue to discard, use 6BV 40% alcoholic solution eluting at last with the flushing of 4BV 20% alcoholic solution, collect eluent, being evaporated to does not have alcohol, and lyophilization promptly gets Radix Notoginseng total arasaponins.Through the HPLC external standard method, Panax Notoginseng saponin R in the refining thing
17%, ginsenoside Rg
138%, ginsenoside Rb
120%, ginsenoside Rd 8%, ginsenoside Re 3%, total saponin content reaches 76%.
Embodiment six
Get pseudo-ginseng granule (3mm-8mm) 500g, 50 ℃ of dryings 6 hours add 12 times of amount 50% ethanol slight boiling condition heating and refluxing extraction 3 times, and each 0.5 hour, extracting liquid filtering merged, and is evaporated to 0.5g/mL (crude drug amount), last D
101Macroporous resin.With 3 times of amount column volume deionized water rinsings, discard, the flushing of reuse 3BV 2% ammonia discards, then with deionized water rinsing to effluent pH=7, continue to discard, use 6BV 50% alcoholic solution eluting at last with the flushing of 4BV 30% alcoholic solution, collect eluent, being evaporated to does not have alcohol, and lyophilization promptly gets Radix Notoginseng total arasaponins.Through the HPLC external standard method, Panax Notoginseng saponin R in the refining thing
17%, ginsenoside Rg
135%, ginsenoside Rb
122%, ginsenoside Rd 6%, ginsenoside Re 3%, total saponin content reaches 73%.
Embodiment seven
Get pseudo-ginseng granule (3mm-8mm) 500g, 50 ℃ of dryings 6 hours add 10 times of amount 70% ethanol slight boiling condition heating and refluxing extraction 2 times, and each 2 hours, extracting liquid filtering merged, and is evaporated to 0.5g/mL (crude drug amount), last D
101Macroporous resin.With 4 times of amount column volume deionized water rinsings, discard, the flushing of reuse 4BV 0.05% ammonia discards, then with deionized water rinsing to effluent pH=7, continue to discard, use 5BV 70% alcoholic solution eluting at last with the flushing of 4BV 30% alcoholic solution, collect eluent, being evaporated to does not have alcohol, and lyophilization promptly gets Radix Notoginseng total arasaponins.Through the HPLC external standard method, Panax Notoginseng saponin R in the refining thing
18%, ginsenoside Rg
144%, ginsenoside Rb
128%, ginsenoside Rd 10%, ginsenoside Re 5%, total saponin content reaches more than 95%.
Embodiment eight: the assay of Radix Notoginseng total arasaponins
Chromatographic conditionChromatographic column Agilent Extend-C
18Post (4.6mm * 250mm, 5 μ m); Adopt gradient elution, mobile phase A is water mutually, and Mobile phase B is acetonitrile mutually, gradient: 0min, 20%B; 30min, 20%B; 55min, 46%B; 65min, 55%B; 70min, 55%B; 72min, 95%B; 80min, 95%B.Flow velocity 1.0mLmin
-1Detect wavelength 203nm; 15 ℃ of column temperatures; Sample size 5 μ L.
The preparation of need testing solutionTake by weighing this product, in volumetric flask, be diluted to scale, shake up, promptly with dissolve with methanol solution.
Assay methodThe accurate need testing solution of drawing injects chromatograph of liquid, measures, promptly.
Embodiment nine: the preparation of drop pill
Get Radix Notoginseng total arasaponins 0.5g and 10.5g Polyethylene Glycol-6000 mix homogeneously of embodiment seven, heating and melting moves in the drop pill drip irrigation behind the change material, and in ℃ liquid paraffin of medicine liquid droplet to 6～8, oil removing makes 300 of drop pill.
Embodiment ten: the preparation of lyophilized injectable powder
Get Lignum Dalbergiae Odoriferae oil 1.5g, join in the saturated hydroxypropyl of 13ml, stirring and dissolving filters, and the filtrate cold drying gets the clathrate powder of Lignum Dalbergiae Odoriferae oil and hydroxypropyl.Except that above-mentioned Lignum Dalbergiae Odoriferae oil closes the clathrate powder of hydroxypropyl, get Radix Notoginseng total arasaponins 0.5g, mannitol 5.5g, calcium disodium edetate 0.9g and the distilled water 2ml of embodiment seven again, behind the said components mixing, lyophilization, 350 of packing, promptly.
Embodiment 11: the preparation of tablet
Get among the embodiment seven Radix Notoginseng total arasaponins an amount of with the microcrystalline Cellulose mix homogeneously, add 3% polyvidone alcoholic solution system soft material, mistake 18 mesh sieve system granules, 60 ℃ of dryings 1 hour, granulate, the adding Pulvis Talci is an amount of, mixing, tabletting, promptly.
Embodiment 12: capsular preparation
Get Radix Notoginseng total arasaponins and Oleum Arachidis hypogaeae semen among the embodiment seven and stir in 2: 18 ratio water dissolving type rustless steel and irritate, add an amount of gelatin and glycerol simultaneously and mix, emit reuse colloid mill defibrination, the mixed liquor that grinds stirs, and makes medicinal liquid; Under 40-50 ℃ of temperature, stir glycerol and distilled water miscible, again with the glycerin liquid temperature to 80-90 ℃, get gelatin and add wherein to mix and stir, become glue until off-bottom, glue was left standstill 4 hours, make offset plate with laminator and use for the pill operation; Above-mentioned medicinal liquid of making and offset plate are sent into the pellet press pill, did cylinder typing in 4 hours at 22-25 ℃ of temperature canyon then, again dry 16-20 hour of 25-30 ℃ of temperature canyon, the qualified soft gelatin capsule of pick, clean with 95% ethanol, dry 4 hours of 25-30 ℃ of canyon, promptly.
Embodiment 13: the preparation of injection
Get among the embodiment seven that Radix Notoginseng total arasaponins is an amount of to be dissolved with 40 ℃ of waters for injection, add an amount of pharmaceutical carrier isotonic agent, the pH value of regulator solution is 7.0-8.0, adds to the full amount of water for injection, and removes thermal source with the ultrafilter ultrafiltration, after measuring pH value, use membrane filtration, after the packing, autoclaving, check, packing, promptly.
Embodiment 14: Radix Notoginseng total arasaponins is to the influence of Acute Myocardial Ischemia in Rats
1, experiment material:
1.1 medicine and reagent
Radix Notoginseng total arasaponins is pressed the preparation of embodiment seven methods.
Verapamil hydrochloride injection, Shanghai Hefeng Pharmaceutical Co., Ltd., lot number: 6E12001.
Lidocaine hydrochloride injection, Zhejiang Cheng Yi Pharmaceutical Co., Ltd, lot number: 20060422.
Chloral hydrate, analytical pure, Chemical Reagent Co., Ltd., Sinopharm Group, lot number: T20060922
TTC, Sigma company
The CK detection kit, Ningbo Asia-Pacific Biotechnology Co.,Ltd, lot number: 06090102.
The LDH detection kit, Ningbo Asia-Pacific Biotechnology Co.,Ltd, lot number: 06060101.
1.2 experiment equipment
The rat oral gavage pin, 2ml syringe, operating theater instruments, toy respirator, thermostat water bath, electronic balance, microplate reader
1.3 laboratory animal
The SD rat, 160-180g, male, Zhejiang Academy of Medical Sciences, licence: SCXK (Zhejiang) 2003-0001.
2, experimental technique
2.1 grouping administration
The 160-180g male SD rat divides at random and does four groups: sham operated rats, model group, positive drug group, Radix Notoginseng group.Positive drug group, lumbar injection give verapamil 0.5mg/kg; The Radix Notoginseng group is irritated stomach and is given Radix Notoginseng total arasaponins 50mg/kg; Sham operated rats and model group are irritated stomach and are given the equal-volume normal saline.Each organized successive administration 7 days.
2.2 model copy
After the 7th administration, each treated animal lumbar injection 300mg/kg chloral hydrate anesthesia.Dorsal position is fixed on the operation plate.Hindlimb muscle injection lignocaine 10mg/kg.Chest medisection skin, 3,4 intercostals are opened breast, extrude heart, ligation anterior descending coronary (sham operated rats threading and not ligation).The ligation point location is the 2mm place under pulmonary conus and left auricle boundary.
2.3 sample collecting
24h after the modeling, lumbar injection 300mg/kg chloral hydrate anesthesia.The postcava blood drawing is for preparation serum.Open breast, take out heart, the normal saline washing is removed blood, the blood vessel of cutting the atrium and linking to each other with ventricle.Ventricle is cut into five of uniform thickness, places 1%TTC, 37 ℃ of water-bath 5min dyeing, and the normal myocardium tissue is dyed brick-red, and slough does not dye, and is canescence.The section of taking-up heart, normal saline is washed unnecessary TTC off, and 10% formalin is fixedly spent the night.
2.4 drug effect evaluation
Detect serum CK, LDH, press the operation of test kit description.The heart section that formaldehyde fixed is spent the night separates and is bolarious normal structure and achromophil slough.Slough, and whole ventricle is weighed calculating infarction rate (%)
The heavy mg/ ventricle of infarction rate %=necrotic myocardium gross weight mg * 100%
3, experimental result
The result shows that model group serum CK, LDH content obviously raise than sham operated rats, and the infarction rate is 20.9 ± 3.5%.The positive drug verapamil can effectively reduce the infarction rate, and serum CK, LDH content.Hints model duplicates successfully.
Be subjected to reagent thing Radix Notoginseng total arasaponins can significantly reduce myocardial infarction rate (15.0 ± 3.1%), with model group comparing difference significance (P＜0.01).Radix Notoginseng group serum CK, LDH content obviously reduce than model group simultaneously, and difference has significance meaning (P＜0.05, P＜0.01).Concrete data see Table 1.
The influence that table 1 Radix Notoginseng total arasaponins (A2) damages Acute Myocardial Ischemia in Rats (X ± SD)
Annotate: compare ##P＜0.01 with sham operated rats; Compare * P＜0.05, * * P＜0.01 with model group
4, conclusion
Damage has significant protective effect to the Radix Notoginseng total arasaponins for preparing by the inventive method to Acute Myocardial Ischemia in Rats, can significantly reduce the myocardial infarction rate.
Embodiment 15
The method disclosed in the present another advantage compared with prior art is and can carries out preferable quality control to Radix Notoginseng total arasaponins.The present invention has set up the analytical method of high performance liquid chromatography, to R
1, Rg
1, Re, Rb
1, the highest monomer saponin of content carries out quantitative analysis in these five kinds of Radix Notoginseng of Rd, the sample separation degree is (referring to accompanying drawing 1) better, satisfies quantitative requirement.Simultaneously, this technology stability and favorable reproducibility are carried out repeated experiments six times with the method for the invention, make six batches of Radix Notoginseng total arasaponinss, carry out liquid phase analysis, and the above five kinds of monomer saponin content RSD of result are all in 5%.
Comprehensive the above, preparing Radix Notoginseng total arasaponins with the method for the invention, to have technology simple, yield is higher, the advantage of favorable reproducibility simultaneously, can be carried out quality control to Radix Notoginseng total arasaponins according to the analytical method that the present invention set up.Table 2 is in the contrast of several respects such as processing step, total saponins assay method and content between the method for the invention and the prior art.
The contrast of table 2 the method for the invention and prior art
| Processing step | The saponin assay method | Saponin content | |
| Patent 1 (CN01108591.6) | Macroporous resin and ion exchange resin have been used, through steps such as extraction, clarification, decolouring, desaccharide, spray dryinges | Spectrophotography is surveyed total saponins, and high-efficient liquid phase technique is surveyed Rg 1，Rb 1，R 1 | Total saponin content 95% (spectrophotometry) |
| Patent 2 (CN93105774.4) | Adopted immersion, pulverizing, lixiviate, fine straining, concentrated, chromatography, decolouring, filtration, concentrate, ten procedures such as drying, use two-step bleaching, two steps dry | Undeclared | Total saponin content 88% |
| Patent 3 (CN02131193.5) | Used quaternary amine base resin | High-efficient liquid phase technique is measured Rg 1，Rb 1，R 1，Re | Total saponin content is more than 60% |
| The present invention | Extraction, purification by macroporous resin, drying | High-efficient liquid phase technique is measured Rg 1，Rb 1，R 1，Re， Rd | Total saponin content is (high-efficient liquid phase technique mensuration) more than 95% |
Claims (2)
1. preparation method of from the Chinese medicine Radix Notoginseng, extracting purified general saponin, its feature realizes by following steps:
(1) pseudo-ginseng was ground into the powder of 20 mesh sieves or particle diameter less than the granule of 1cm, removed impurity, and oven dry is with containing the water-soluble matchmaker's diafiltration of ethanol that concentration is 0-95%, ultrasonic, hot reflux is extracted, and solvent load 5-20 doubly measures, extract 1-3 time, each 0.5-2 hour, the enrichment of extracting solution concentrating under reduced pressure;
(2) D on the concentrated solution
101Macroporous resin, successively with water, ammonia, pure water elution, 4-6 column volume of elder generation's water flushing, 4-6 column volume of reuse 0.05%-1% ammonia flushing, reuse water washes to pH=7,3-5 column volume of reuse 20-30% alcohol flushing, use 3-6 column volume of 60-80% ethanol elution at last, collect eluent, concentrate, drying reclaims solvent and promptly gets the refining thing of Radix Notoginseng total arasaponins.
2. a kind of purified general saponin preparation method of extracting from the Chinese medicine Radix Notoginseng according to claim 1 is characterized in that: D on the sample in the step (2)
101Macroporous resin, 4-5 column volume of elder generation's water flushing, 4-5 column volume of reuse 0.05%-0.1% ammonia flushing, reuse water washes to pH=7,4-5 column volume of reuse 20-30% alcohol flushing, use 5-6 column volume of 60-70% ethanol elution at last, collect eluent, reclaim solvent and get the refining thing of Radix Notoginseng total arasaponins.
Priority Applications (1)
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| CN101463061B (en) * | 2007-12-21 | 2013-09-18 | 中国医学科学院药物研究所 | Ginseng saponin Rg1 and Rb1 in pseudo-ginseng and preparation of total saponin thereof |
| CN102166238B (en) * | 2011-04-08 | 2012-06-20 | 桂林英美特生物技术有限公司 | Notoginseng extract and application thereof |
| CN103462009A (en) * | 2013-09-04 | 2013-12-25 | 云南七丹药业股份有限公司 | Traditional Chinese medicine extract for assisting reduction of blood fat and preparation method thereof |
| CN104697950A (en) * | 2015-03-16 | 2015-06-10 | 刘旭 | Method for detecting pseudo-ginseng content and optimally extracting and purifying |
| CN104800258A (en) * | 2015-04-09 | 2015-07-29 | 广西大学 | Method for preparing panax notoginseng saponins |
| CN105125609B (en) * | 2015-09-30 | 2019-01-15 | 桂林三宝药业有限公司 | A method of extracting notoginseng total saponin from Radix Notoginseng |
| CN106176864B (en) * | 2016-08-23 | 2017-06-16 | 李续博 | It is a kind of to treat panax notoginseng saponin medicine composition of scar and preparation method thereof |
| CN106420871B (en) * | 2016-09-28 | 2020-06-19 | 澳门大学 | Preparation method of total notoginsenoside rich in Fe and Fd and monomers Fe and Fd thereof |
| CN107693559A (en) * | 2017-11-27 | 2018-02-16 | 贵州浩诚药业有限公司 | The preparation method of Notogineng Extract |
| CN112010911A (en) * | 2019-05-31 | 2020-12-01 | 泰州医药城国科化物生物医药科技有限公司 | Method for purifying total ginsenoside |
| CN113116949A (en) * | 2020-01-15 | 2021-07-16 | 四川森科制药有限公司 | Extraction process of panax notoginseng saponins |
| CN114432324A (en) * | 2021-12-13 | 2022-05-06 | 吉林省道地参茸有限公司 | New application of triterpenoid saponin compound in treating hyperuricemia and gout |
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