CN101392013A - Ginsenoside Rb1 containing impurity ginsenoside Rh1 - Google Patents
Ginsenoside Rb1 containing impurity ginsenoside Rh1 Download PDFInfo
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Abstract
The invention discloses a standard ginsenoside Rb1 active ingredient, which contains ginsenoside Rb1 with the content more than or equal to 90 percent and smaller than 100 percent, and is characterized in that the content of impurity ginsenoside Rh1 is more than 0 and smaller than 5 percent; or the content of the impurity ginsenoside Rh1 is more than or equal to 0.1 percent and smaller than 5 percent; or the content of the impurity ginsenoside Rh1 is more than or equal to 0.5 percent and smaller than 3.0 percent; the pharmacological test shows that the active ingredient of the application has good pharmacological action, and can be prepared to pharmaceutical preparation in pharmacy.
Description
Technical field
The present invention relates to technical field of traditional Chinese medicines, be specifically related to a kind of ginsenoside Rb of standard
1Effective constituent.
The invention reside in the middle pharmaceutically active ingredient that a kind of standard is provided, promptly contain impurity ginsenoside Rh
1The ginsenoside Rb of standard
1
Background technology
Ginsenoside has good pharmacologically active, is divided into olea acids, panoxadiol class, panoxatriol class by the aglycon difference, and that have fine pharmacologically active is Panaxadiol saponin, Panaxatriol saponin, and monomer saponin has-R
0, RA
0, Ra
1,-Ra
2,-Ra
3,-Rb
1,-Rb
2,-Rb
3,-Rc ,-Rd ,-Re ,-Rg
1,-Rg
2,-Rg
3,-Rh
1,-Rh
2Deng, Panaxadiol saponin is with ginsenoside Rb
1Be main, great pharmacological effects is arranged: as: the panoxadiol saponins is to rat myocardial cell calcium channel blocking action (preclinical medicine and clinical, 1994,02), the panoxadiol monomer is to the single calcium channel analysis and the ESR spectrum research (preclinical medicine and clinical of rat heart muscle effect, 1994,04), the panoxadiol saponins is to myocardial ischemia-reperfusion dog CO, the influence of TPVR and serum N E content (Norman Bethune Medical University's journal 1995,05), Panaxadiol saponin is to provide protection (the Chinese Pharmacological circular of pallasiomy acute cerebral ischemia-reperfusion injury, 1996,06), the panoxadiol saponins is to influence (Chinese Chinese materia medica science and technology 200403) of rabbit ischemical reperfusion injury cardiac hemodynamic etc.; Ginsenoside Rb
1Be mainly derived from Araliaceae Panax genseng, pseudo-ginseng, Radix Panacis Quinquefolii, Vietnam's genseng, rhizome of Japanese Ginseng, in the root of plants such as Curcurbitaceae gynostemma pentaphyllum genus gynostemma pentaphylla, edge fruit gynostemma pentaphylla, stem, leaf, the bud, the Chinese medicinal materials of using different genera extracts the ginsenoside Rb that purifying obtains
1Impurity be different certainly, therefore, should at ginsenoside Rb
1Impurity carry out deep research, the research of impurity in the middle pharmaceutically active ingredient is clear, particularly that content is bigger impurity is studied and quantitatively control, help us clearly to understand the basic substance of effective constituent, after the medicine listing, when untoward reaction or side effect take place, help us and effectively follow the trail of root, effectively impurity or synergy impurity should quantitatively be controlled, invalid impurity should carry out content and limit, poisonous impurity or detrimental impurity should further be removed, and perhaps content is limited to harmless scope (such as less than 10PPM), therefore, impurity in the centering pharmaceutically active ingredient carries out deep research, have profound significance, help the modernization of Chinese medicine, help Chinese medicine to go to the world, help the healthy of the mankind more.
Ginsenoside Rb
1Many employing laboratory methods obtain a small amount of (the mg level is to the g level), such as methods such as preparative high-performance liquid chromatographic method, silica gel column chromatographies, are used for scientific research, but can't obtain (more than the kg level) ginsenoside Rb in batches
1Be used for patent medicine exploitation and suitability for industrialized production, so this is ginsenoside Rb
1So far the reason and the difficult point that are difficult to patent medicine.
Application number is 03127664.4 patent documentation " environment-friendly type ginsenoside Rb
1Height obtain volume production already divide from ", the document discloses and can obtain content greater than 95% ginsenoside Rb
1, adopting silica gel column chromatography method, this method involves great expense, and is difficult to suitability for industrialized production; Another subject matter of the document is not have the method for analyzing and testing to show ginsenoside Rb
1Content reach 95%, its confidence level be worth to be suspected; Application number is patent documentation " the ginsenoside Rb of 03148803.x
1Preparation technology ", the method that the document discloses the method+silica gel column chromatography of employing Amberlyst process+silica gel column chromatography obtains ginsenoside Rb
1, this method is only applicable to laboratory study equally, is difficult to realize suitability for industrialized production; Another subject matter of the document is to obtain ginsenoside Rb
1Do not have purity, do not have the method for assay yet; The patent documentation of application number " 200610093610.6 " method of extraction separation ginsenoside monomer " a kind of from the Ginseng Leaf ", the document discloses the method for employing Amberlyst process+silica gel column chromatography and has separated the method that obtains different ginsenoside monomers, comprises ginsenoside Rb
1Its subject matter does not still have the requirement of purity, does not have content assaying method, and this processing method still adopted this complexity of column chromatography, cost height, can't suitability for industrialized production method.
In sum, obtaining ginsenoside Rb
1On the basis of effective constituent, study, obtain standard active ingredient, have profound significance at its impurity impurity that particularly content is bigger.
Summary of the invention
For these reasons, our scientific research personnel obtains ginsenoside Rb to the medicinal material of different genera, different methods
1Carry out deep analysis, in that " content is greater than 90% ginsenoside Rb
1, total impurities is smaller or equal to 10% " the basis on, by performing creative labour, proved conclusively ginsenoside Rb
1In content bigger one of impurity be the ginsenoside Rh
1, because, the ginsenoside Rh
1Have very strong hemolytic action, belonging to promptly is that effective constituent is again objectionable constituent, and therefore, we are to the ginsenoside Rh
1The control of limiting the quantity of obtains standard ginsenoside Rb
1Effective constituent has promptly kept impurity ginsenoside Rh
1Pharmacologically active, guaranteed it again within the specific limits, hemolytic action is reduced to minimum, this standardized ginsenoside Rb
1Effective constituent, can be directly as medicine material in market sale, the preparation raw material that also can be used as the medicament preparation uses, and like this, helps the stdn of Chinese medicine material, helps the stdn that Chinese medicine preparation is produced; Simultaneously, our scientific research personnel is also at ginsenoside Rb
1Extraction and purification process study: adopting Amberlyst process, is raw material with the total saponins, separates obtaining ginsenoside Rb
1Crude product by recrystallization method, obtains ginsenoside Rb again
1, this processing method is simple, can realize suitability for industrialized production, the ginsenoside Rb that obtains
1Cost is not high, its content and impurity is analyzed ginsenoside Rb
1Content is more than or equal to 90%, ginsenoside Rh in the impurity
1Content is less than 5%, conformance with standard ginsenoside Rb
1The requirement of effective constituent; The application is to containing impurity ginsenoside Rh
1Ginsenoside Rb
1Carry out pharmacologically active experiment, find that it except that having effect such as good treatment cardiovascular and cerebrovascular diseases, also has good antineoplastic action, with existing ginsenoside Rb
1(do not contain the ginsenoside Rh
1Or content is extremely low) relatively, aspect antitumor action, have significant difference (P＜0.05).
The present invention is achieved through the following technical solutions.
The application is to provide a kind of ginsenoside Rb of standard
1, i.e. the middle pharmaceutically active ingredient of standard: effective constituent is carried out content determine, impurity is carried out material is determined and content is determined; The application's standard ginsenoside Rb
1Effective constituent can be used as the pharmaceutical preparation bulk drug and uses, on the basis that meets the pharmaceutical preparation requirement, and the ginsenoside Rb of preparation
1Pharmaceutical preparation can be used for the treatment of human body diseases.
The ginsenoside Rb of the application's standard
1Effective constituent is:
(1) a kind of ginsenoside Rb
1, ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, and foreign matter content is characterized in that greater than 0 and smaller or equal to 10% impurity comprises the ginsenoside Rh
1, Rh
1Content greater than 0 and less than 5%.
(2) a kind of ginsenoside Rb
1, ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, wherein impurity ginsenoside Rh
1Content more than or equal to 0.1% and less than 5%.
(3) a kind of ginsenoside Rb
1, ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, wherein impurity ginsenoside Rh
1Content more than or equal to 0.5% and smaller or equal to 3.0%.
Ginsenoside Rh in the above-mentioned standard active ingredient
1For content in the impurity is one of bigger, by scientific experiment it is separated, determine its structure by serial experiment again.
Above-mentioned standard active ingredient can be by genseng, pseudo-ginseng, Radix Panacis Quinquefolii, Vietnam's genseng, rhizome of Japanese Ginseng, obtain total saponins in the root of plants such as Curcurbitaceae gynostemma pentaphyllum genus gynostemma pentaphylla, edge fruit gynostemma pentaphylla, stem, leaf, the bud, carrying out purifying through the high performance liquid chromatography preparation method again obtains, also can carry out monomer separation by silica gel column chromatography, high performance liquid chromatography control terminal point obtains, or the like;
The above-mentioned standard ginsenoside of the application Rb
1Effective constituent also can be obtained by following method:
(1) gets pseudo-ginseng, genseng, Radix Panacis Quinquefolii or gynostemma pentaphylla and extract the total saponins that obtains;
(2) total saponins is dissolved in water, and filters, and filtrate is passed through nonpolar or low-pole macroporous adsorptive resins, washing, 40% ethanol elution, and elutriant discards, and uses 50% ethanol elution again, collects 50% elutriant, drying;
(3) dry thing filters with ethanol, 95% ethanol, propyl carbinol or dissolve with methanol, and filtrate adds or do not add organic solvent, and placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Total saponins in the above-mentioned steps 1 can directly be bought by market or prepare according to scientific and technical literature;
The ginsenoside Rb that above-mentioned steps (3) obtains
1Method that can repeating step (3);
Described Radix Notoginseng total arasaponins can perhaps prepare (Wei Junxian, Chen Yegao, Cao Shuming, the research of pseudo-ginseng carpopodium saponin component, CHINA JOURNAL OF CHINESE MATERIA MEDICA, 1992,17 (9): 611-613 according to existing patent documentation or scientific and technical literature by commercially available purchase; Yang Chongren, kingdom swallow, 5 jewels, etc., the saponin component of pseudo-ginseng reed head, pharmacy circular, 1985,20 (6): 337-338; Dong Aling, Zheng Junhua, fruit Dean etc., the method for from pseudo-ginseng, separating the trace ingredients arasaponin, CHINA JOURNAL OF CHINESE MATERIA MEDICA, 2002,27 (10): 793-794; Ou Lailiang, Shi Zuoqing, Shi Rongfu, etc., the epistasis macroporous adsorbent resin is to the separation and purification research of arasaponin, herbal medicine, 2003,34 (10): 905-907; Zhang Guande, adsorption resin method is measured pseudo-ginseng and the total saponin of preparation perhexiline thereof, herbal medicine, 1981,12 (11): 23-25.).
Described macroporous resin column is HPD-100, HPD-100A, HPD-300, HPD-400, HPD-400A, HPD-450, D101,1300-I, 1400 or AB-8.
Described organic solvent is one or both in acetone, the ethyl acetate.
Above-mentioned ginsenoside Rb
1Pharmaceutical preparation for the unitary dose of active substance preparation.
Above-mentioned ginsenoside Rb
1Be the pharmaceutical preparation of active substance preparation, wherein unitary dose is 1mg-4000mg.
Above-mentioned ginsenoside Rb
1Be the pharmaceutical preparation of active substance preparation, wherein unitary dose is 5mg-2000mg.
Above-mentioned ginsenoside Rb
1Treat and/or prevent application in cardiovascular and cerebrovascular diseases, tumour, aging, the hypomnesis medicine in preparation.
One, detection method
Experimental technique:
Adopt high performance liquid chromatography to carry out check and analysis;
Adopt high performance liquid chromatography to carry out check and analysis; Chromatographic column: C
18Reverse-phase chromatographic column; Chromatographic condition and system suitability experiment: with the octadecylsilane chemically bonded silica is weighting agent; Flow velocity 1.0ml/min; 25 ℃ of column temperatures; Detect wavelength 203nm; Number of theoretical plate is pressed ginsenoside Rb
1Meter should be not less than 2500; With water: acetonitrile is a moving phase, carries out gradient elution by following condition of gradient elution, moves 60 minutes;
In the time of 0-40 minutes, the ratio of water reduces to 50% by 80%, and the ratio of nitrile rises to 50% by 20%; In the time of 40-43 minutes, the ratio of water reduces to 20% by 50%, and the ratio of nitrile rises to 80% by 50%; In the time of 43-48 minutes, keeping the ratio of water is 20%, and keeping, the ratio of nitrile is 80%; In the time of 48-50 minutes, the water ratio rises to 80% by 20%, and the ratio of water reduces to 20% by 80%; 50-60 minutes keep the ratio of water is 20%, the ratio 80% of nitrile;
The preparation of reference substance solution: precision takes by weighing ginsenoside Rb
1Reference substance adds dissolve with methanol and shakes up in volumetric flask, and is diluted to scale;
The preparation of need testing solution: precision takes by weighing or measures sample and adds dissolve with methanol in the volumetric flask and shake up, and is diluted to scale;
Assay method: accurate respectively absorption reference substance solution and need testing solution respectively inject liquid chromatograph, adopt by external standard method with calculated by peak area, promptly.
Annotate: the application's genseng saponin(e Rb
1Reference substance is the statutory standards product, buys from Chinese pharmaceutical biological product evaluation and buys.
The application's standard ginsenoside Rb
1The check and analysis of effective constituent
Get the application's standard ginsenoside Rb
1Effective constituent, carry out the check and analysis experimental result according to above-mentioned experimental technique and see Table 1:
Table 1 ginsenoside Rb
1The sample determination result
Experiment conclusion: show the application's standard ginsenoside Rb by above-mentioned experiment
1In the effective constituent, ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, impurity ginsenoside Rh
1Content greater than 0 and less than 5%; Ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, impurity ginsenoside Rh
1Content more than or equal to 0.1% and less than 5%; Ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, impurity ginsenoside Rh
1Content more than or equal to 0.5% and smaller or equal to 3%.
Two, ginsenoside Rh
1The structural identification data
Get the application's standard ginsenoside Rb
1Effective constituent is separated through silica gel column chromatography, in conjunction with high performance liquid chromatography liquid method, and the impurity that content is bigger, ginsenoside Rb
1Separate respectively, obtain the impurity monomer, carry out structural identification, the conclusive evidence data is as follows:
13C-NMR: see Table 2
Table 2 ginsenoside Rh
1The structural identification data
1H-NMR：
1 .949,1.000,1.010,1.096,1.158,1.332,1.627,1.689 exist the CH in 8 methyl peak-to-peak signal counter structures
3-18,19,21,26,27,28,29,30.
2, δ 5.146, and t is the fignal center of CH-24.
3, δ 4.350, d, J=7.6 by 1 carbon of glucose the fignal center of the H of company.
Experiment conclusion: show that by above-mentioned experiment one of big impurity of the application's content is the ginsenoside Rh
1
Three, hemolytic experimental study
Experimental program:
Scheme 1: ginsenoside Rb
1Content 90.02%, impurity ginsenoside Rh
1Content 9.14%.
Scheme 2: ginsenoside Rb
1Content 91.89%, impurity ginsenoside Rh
1Content 7.76%.
Scheme 3: ginsenoside Rb
1Content 92.57%, impurity ginsenoside Rh
1Content 6.33%.
Scheme 4: ginsenoside Rb
1Content 93.18%, impurity ginsenoside Rh
1Content 5.27%.
Scheme 5: ginsenoside Rb
1Content 94.10%, impurity ginsenoside Rh
1Content 5.02%.
Scheme 6: ginsenoside Rb
1Content 94.27%, impurity ginsenoside Rh
1Content 4.99%.
Scheme 7: ginsenoside Rb
1Content 95.31%, impurity ginsenoside Rh
1Content 3.87%.
Scheme 8: ginsenoside Rb
1Content 96.72%, impurity ginsenoside Rh
1Content 3.00%.
Scheme 9: ginsenoside Rb
1Content 97.03%, impurity ginsenoside Rh
1Content 1.95%.
Scheme 10: ginsenoside Rb
1Content 98.31%, impurity ginsenoside Rh
1Content 0.50%.
Scheme 11: ginsenoside Rb
1Content 99.24%, impurity ginsenoside Rh
1Content 0.10%.
Scheme 12: ginsenoside Rb
1Content 99.87%, impurity ginsenoside Rh
1Content 0.07%.
Scheme 13: physiological saline.
Experimental technique: remove the scleroproein rabbit whole blood, add physiological saline, shake up, centrifugal, the supernatant liquor that inclines is washed clearly till do not become redness repeatedly, measures red corpuscle, adds physiological saline and is diluted to 2% suspension, gets the ginsenoside Rb of above-mentioned different schemes
1, add 2% red blood cell suspension 2.5ml, add physiological saline to 5ml, shake up gently, put in 37 ℃ of waters bath with thermostatic control, observe 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours haemolysis situations, experimental result sees Table 3.
The haemolysis situation of table 3 different schemes
Annotate :+expression haemolysis ,-represent not haemolysis ,+-expression has haemolysis but not exclusively
Experimental result: show by the hemolytic experimental study, work as impurity ginsenoside Rh
1Content did not almost have hemolytic action less than 5% o'clock, and the impurity ginsenoside Rh that contains is described
1Should be limited to greater than 0 and less than in 5% the scope.
Four, pharmacological evaluation
Experiment 1
Comparison to the rats'liver tumor suppression
Laboratory animal: rat, 150g-180g, male and female are regardless of.
Experiment medicine: physiological saline; The application's standard ginsenoside Rb
1Effective constituent (meets the application's standard active ingredient to impurity ginsenoside Rh
1Qualification); Commercially available ginsenoside Rb
1Monomer (Chinese pharmaceutical biological product is identified institute); Sodium cantharidin injection liquid (Guizhou Shenqi Pharmaceutical Co., Ltd)
Experimental technique: get rat and be divided into physiological saline group, commercially available sodium cantharidin injection liquid group, the application's standard ginsenoside Rb
1Effective constituent group, commercially available ginsenoside Rb
1Set of monomers is made W
256Liver in inoculation, inoculate after 7 days, press the dosage intraperitoneal injection of anesthesia of 35mg/kg with vetanarcol, fixing, cutting open the belly exposes liver, tumor surface maximum diameter (a) and path (b) are pressed (a*b on the measurement liver
2)/2=V (gross tumor volume).Separate stomach, arteria duodenalis, arteria hepatica communis and proper hepatic artery, ligation stomach, arteria duodenalis far-end, with silver brain clip blocking-up arteria hepatica communis, in sending into proper hepatic artery again at the gastroduodenal artery upper cut and after inserting external diameter 0.3mm conduit under the operating loupe, inject respectively by the experiment grouping then and be subjected to reagent thing (dosage is identical), postoperative tube drawing ligation gastroduodenal artery, decontrol the arteria hepatica communis silver brain clip, sew up the incision again, place animal housing to wait to revive rat, continue breeding observing, performed the operation back 8 days, detect gross tumor volume by last method, experimental result sees Table 4:
Table 4 is respectively organized preparation to the rejection ratio of tumour
Annotate: compare with the physiological saline group
*P＜0.01; Compare #P＜0.05 with the commercially available sodium cantharidin injection liquid of positive controls group
Experiment 2
Physics, chemistry and biotic factor all can be brought out cancer, but the 80%-90% of people's cancer pathogenic factor thinks what the environmental chemistry material caused.
Diethylnitrosamine (DENA) is brought out the inhibition of rat liver cancer
Laboratory animal: rat, about 120g, male and female are regardless of.
Experiment medicine: physiological saline; The application's standard ginsenoside Rb
1Effective constituent (meets the application's standard active ingredient to impurity ginsenoside Rh
1Qualification); Commercially available ginsenoside Rb
1Monomer (Chinese pharmaceutical biological product is identified institute); Sodium cantharidin injection liquid (Guizhou Yibai Pharmaceutical Co., Ltd)
Experimental technique: feed rat with the drinking-water that contains diethylnitrosamine (DENA), 36 all primary hepatocarcinoma that get experimentize according to the method for testing 1, record survival of rats fate, and experimental result sees Table 5:
Table 5 survival of rats fate relatively
Annotate: compare with the physiological saline group
*P＜0.01; Compare #P＜0.05 with the commercially available sodium cantharidin injection liquid of positive controls group
Experiment 3
Provide protection to rat cerebral infarction
Experiment medicine: the application's standard ginsenoside Rb
1Effective constituent (meets the application's standard active ingredient to impurity ginsenoside Rh
1Qualification); Commercially available ginsenoside Rb
1Monomer (Chinese pharmaceutical biological product is identified institute); XUESHUANTONG ZHUSHEYE.
Laboratory animal: SD rat, 250-300g, male and female half and half.
Experimental technique: get rat, Chloral Hydrate 300mg/kgip anesthesia, cervical incision, separate and the ligation right carotid, behind the suture muscles skin, right lateral position is fixed, and cuts skin at auris dextra and right eye outer canthus line mid point, separate temporalis, expose zygomatic process and temporal bone, open the bone window of about a 3 * 3mm at head end 1～2mm place of zygomatic process, expose arteria cerebri media (MCA), it is disconnected that MCA is burnt, and sews up the incision.With reference to the administration of Bederson method, put to death animal, get right cerebral hemisphere, be cut into 5, place 37 ℃ of incubation 15min dyeing of 2g/LNPT, the white infarct cerebral tissue that carefully takes and weigh and do not dyed blue look, experimental result sees Table 6.
Table 6 different pharmaceutical is protected situation to rat cerebral infarction
Annotate: compare with the physiological saline group
*P＜0.01; Compare #P＜0.05 with the XUESHUANTONG ZHUSHEYE group
Experiment 4
To the effect of dog ischemic myocardial protection
Laboratory animal: Beagle dog, body weight 7.5-10kg.
Experiment medicine: the application's standard ginsenoside Rb
1Effective constituent; Commercially available ginsenoside Rb
1Monomer (Chinese pharmaceutical biological product is identified institute); XUESAITONG ZHUSHEYE.
Experimental technique: get experimental dogs, be divided into physiological saline group, XUESAITONG ZHUSHEYE group, the application's standard ginsenoside Rb
1Effective constituent group, commercially available ginsenoside Rb
1Set of monomers, the administration group is respectively in the subcutaneous cephalic vein administration of dog forelimb, the each 20mg/kg of dosage, physiological saline groups etc. are held inequality, administration every day 1 time, successive administration 3 days, administration are used vetanarcol 25mg/kg intravenous anesthesia, trachea cannula after the 3rd day, connect breathing apparatus's positive pressure respiration in addition, the 5th rib is opened chest from the left side, cuts off pericardium, exposes heart, the pericardial incision edge is sewn in the wall of the chest, divide the second stage of ligation at ramus descendens anterior arteriae coronariae sinistrae, and slow iv lignocaine 8mg/kg is in case the ventricular fibrillation that may cause before the ligation is sewed up the pericardium and the wall of the chest subsequently.Heart stalk back 24h, dog is put to death after vetanarcol anesthesia, take out heart rapidly, excision great vessels and fatty tissue, check the position of descending anterior branch ligation point, claim heavy whole-heartedly, excise right ventricle and left atrium then, stay interventricular septum and left atrium, claim to such an extent that left ventricle is heavy, from the apex of the heart and beginning and descending anterior branch vertical direction left ventricle is cut into the thick flesh sheet of 2-3mm, be dipped in dyeing 30min in the nitro ditetrazolium chloride solution (NBT), cut ischemic region cardiac muscular tissue and weigh, the calculating myocardium infarction size, that is: heart stalk scope=ischemic region weight/left ventricle is heavy by * 100%, the results are shown in Table 7.
The influence of table 7 pair dog ventricle infarction size
Annotate: compare with the physiological saline group,
*P＜0.01; Compare #P＜0.05 with positive controls XUESAITONG ZHUSHEYE group
Annotate: with the application's standard ginsenoside Rb
1Effective constituent and commercially available ginsenoside Rb
1Monomer carries out experiment, dementia resisting experiment, the sexual function improving experiment of mouse memory function, and experimental result shows, the application's standard ginsenoside Rb
1Effective constituent is than commercially available ginsenoside Rb
1The monomer pharmacologically active is good.
Discuss: show that by above-mentioned pharmacological evaluation the application contains impurity ginsenoside Rh
1Ginsenoside Rb
1Ratio content is near 100% ginsenoside Rb
1The monomer pharmacologically active will be got well, and through our research, preliminary interpretation is: ginsenoside Rh1 also is an activeconstituents, also is ginsenoside Rb simultaneously
1Degraded product, the two except that having summation action, also can have synergy in entering body, in addition, contain the ginsenoside Rb of a certain amount of impurity ginsenoside Rh 1
1Aspect drug metabolism, can promote ginsenoside Rb1's absorption, and content is near 100% ginsenoside Rb
1May be not contain ginsenoside Rh1 or content trace, to ginsenoside Rb
1Influence very little, can not produce synergy (aspect such as pharmacology and medicine generation), therefore, its pharmacologically active is than the application's standard ginsenoside Rb
1The pharmacologically active of effective constituent is low.
Four, dose screening experiment
To mouse S
180The tumor growth restraining effect
Laboratory animal: healthy mice, body weight 16-20g, male and female half and half.
Experimental technique: get and inoculate the mouse S that goes down to posterity
180, in homogenizer, add physiological saline, make mouse S
180The knurl homogenate, again with physiological saline 1:3 dilution, getting 0.2ml then, to inject oxter, a mouse left side subcutaneous, weighed in 24 hours, mouse tail every day intravenously administrable once, physiological saline etc. hold inequality, totally 7 days.Next day is put to death mouse in drug withdrawal, and the subcutaneous tumors piece is peeled off in the also carefulness of weighing, and takes by weighing knurl in the EM50 electronic balance and weighs, and calculate tumour inhibiting rate, sees Table 8:
Table 8 different dosing dosage is to mouse S
180The tumor growth restraining effect
Annotate: compare with positive controls KANGLAITE ZHUSHEYE group
*P＜0.01, * P＜0.05.
Experiment conclusion: show the application's genseng saponin(e Rb by the dose screening experiment
1Increasing pharmacologically active with dosage also strengthens, it is different with the pharmacologically active of a lot of medicines that (pharmacologically active can arrive a plateau to most of medicine with the dosage increase, be that dosage increases again and activity does not increase), along with increasing activity, dosage increases always, do not have plateau, can arrive always and occur till the toxicity; Through us dose screening experiment and toxicity test, determine the application's genseng saponin(e Rb
1Unitary dose be 1mg-4000mg; Preferred unit dosage is 5mg-2000mg (dosage toxic reaction occurs greater than 4000mg, does not have significant difference less than 1mg and model group).
The application's standard ginsenoside Rb
1Effective constituent and prior art ginsenoside Rb
1(" application number is patent documentation＜environment-friendly type ginsenoside Rb of 03127664.4
1Height obtain volume production already divide from, the document discloses and can obtain content greater than 95% ginsenoside Rb
1" etc. method obtain) relatively have following characteristics:
1, the application is with ginsenoside Rb
1The ownership that one of impurity that middle content is bigger carries out has determined that it is the ginsenoside Rh
1, further clear and definite basic substance, and just obtained the ginsenoside Rb of certain content in the prior art
1, not carrying out deep research, basic substance is fuzzy;
2, the application marks ginsenoside Rb
1Effective constituent can be used as the raw material of pharmaceutical preparation, in clinical application, any untoward reaction or side effect take place, because basic substance is clearer and more definite, particularly impurity clearly can be in the research of follow-up clinical drug, go out clearly that the composition of medicine own causes, or the no related substance of introducing in the preparation of pharmaceutical formulations process (such as factors such as thermal source, pollutions) causes, help the development of medicine, and ginsenoside Rb of the prior art
1Though be effective constituent, also exist certain camera bellows, when untoward reaction or side effect particularly take place, can't demand origin, cause the obstacle (such as the Herba Houttuyniae injectio incident) of drug development;
3, the application determines impurity ginsenoside Rh under study for action
1Belong to be effective impurity be again poisonous impurity, and define its scope, on certain basis to effective constituent ginsenoside Rb
1Have synergy, belong to effective impurity, and the prior art ginsenoside Rb that has been clear and definite
1Content, all the other materials are belonged to effective impurity, invalid impurity or detrimental impurity know nothing, research belongs to initial period.
Four, embodiment
Embodiment 1
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 90.01%, impurity ginsenoside Rh
1Content 4.99%.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 2
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 99.9%, impurity ginsenoside Rh
1Content 0.03%.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 3
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 94.82%, impurity ginsenoside Rh
1Content 4.10%.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 4
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 98.93%, impurity ginsenoside Rh
1Content 0.10%.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 5
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 96.11%, impurity ginsenoside Rh
1Content 3.00%.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 6
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 96.19%, impurity ginsenoside Rh
1Content 0.50%.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 7
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 98.26%, impurity ginsenoside Rh
1Content 0.32%.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 8
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 97.30%, impurity ginsenoside Rh
1Content 1.19%.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 9
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 97.09%, impurity ginsenoside Rh
1Content 1.83%.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 10
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 96.54%, the content 2.68% of impurity ginsenoside Rh 1.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Ginsenoside Rh in the foregoing description 1-10 standard active ingredient
1For content in the impurity is one of bigger, by scientific experiment it is separated, determine its structure by serial experiment again.
The foregoing description 1-10 standard active ingredient can be by genseng, pseudo-ginseng, Radix Panacis Quinquefolii, Vietnam's genseng, rhizome of Japanese Ginseng, obtain total saponins in the root of plants such as Curcurbitaceae gynostemma pentaphyllum genus gynostemma pentaphylla, edge fruit gynostemma pentaphylla, stem, leaf, the bud, carrying out purifying through the high performance liquid chromatography preparation method again obtains, also can carry out monomer separation by silica gel column chromatography, high performance liquid chromatography control terminal point obtains, or the like;
Embodiment 11
Get Radix Panacis Quinquefolii and extract the American ginseng total saponins obtain and be dissolved in water, filter, filtrate is by the HPD-100A macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collect 50% elutriant, drying, dry thing dissolve with ethanol filters, filtrate adds acetone, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
The method that Radix Panacis Quinquefolii extracts total saponins is prepared according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 12
Get pseudo-ginseng and extract the Radix Notoginseng total arasaponins obtain and be dissolved in water, filter, filtrate is by the HPD-100 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collect 50% elutriant, drying, dry thing 95% dissolve with ethanol filters, filtrate adds the organic solvent ethyl acetate, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins extracting method is prepared according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 13
Get gynostemma pentaphylla and extract and to obtain total saponins and be dissolved in water, filter, filtrate is by the HPD-400 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collect 50% elutriant, drying, dry thing dissolves with propyl carbinol, filters, filtrate adds organic solvent-acetone, ethyl acetate, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described gynostemma total saponin extracting method is prepared according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 14
Get genseng and extract the total saponins obtain and be dissolved in water, filter, filtrate is by the HPD-300 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collect 50% elutriant, drying, dry thing dissolve with methanol filters, filtrate adds organic solvent-acetone, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
The extracting method of described Radix Ginseng total saponins can lead to the existing patent documentation of basis or scientific and technical literature is prepared.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 15
Get American ginseng total saponins and be dissolved in water, filter, filtrate is by the HPD-450 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collect 50% elutriant, drying, dry thing dissolve with ethanol filters, filtrate adds the organic solvent ethyl acetate, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described American ginseng total saponins prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 16
Get Radix Notoginseng total arasaponins and be dissolved in water, filter, filtrate is by the HPD-400A macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collect 50% elutriant, drying, dry thing 95% dissolve with ethanol filters, filtrate adds organic solvent-acetone, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins prepares (Zhang Guande, adsorption resin method mensuration pseudo-ginseng and the total saponin of preparation perhexiline thereof, herbal medicine, 1981,12 (11): 23-25.) according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 17
Get Radix Notoginseng total arasaponins and be dissolved in water, filter, filtrate is by the 1300-I macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collect 50% elutriant, drying, dry thing dissolves with propyl carbinol, filters, filtrate adds organic solvent-acetone, ethyl acetate, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins is by commercially available purchase.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 18
Get Radix Notoginseng total arasaponins and be dissolved in water, filter, filtrate is by 1400 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collect 50% elutriant, drying, dry thing dissolve with methanol filters, filtrate adds the organic solvent ethyl acetate, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins according to existing patent documentation or scientific and technical literature prepare (Yang Chongren, kingdom swallow, 5 jewels, etc., the saponin component of pseudo-ginseng reed head, pharmacy circular, 1985,20 (6): 337-338).
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 19
Get Radix Panacis Quinquefolii and extract the total saponins obtain and be dissolved in water, filter, filtrate is by the D101 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collect 50% elutriant, drying, dry thing 95% dissolve with ethanol filters, filtrate adds the organic solvent ethyl acetate, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described American ginseng total saponins extracting method prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 20
Get Radix Ginseng total saponins and be dissolved in water, filter, filtrate is by the AB-8 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collect 50% elutriant, drying, dry thing 95% dissolve with ethanol filters, filtrate adds organic solvent-acetone, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described Radix Ginseng total saponins prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 21
Get gynostemma pentaphylla and extract and to obtain total saponins and be dissolved in water, filter, filtrate is by 1400 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collect 50% elutriant, drying, dry thing dissolves with propyl carbinol, filters, filtrate adds the organic solvent ethyl acetate, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described gynostemma total saponin extracting method prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 22
Get pseudo-ginseng and extract and to obtain total saponins and be dissolved in water, filter, filtrate is by the D101 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collect 50% elutriant, drying, dry thing dissolve with methanol filters, filtrate adds the organic solvent ethyl acetate, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 23
Get Radix Notoginseng total arasaponins and be dissolved in water, filter, filtrate is by the HPD-300 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collect 50% elutriant, drying, dry thing dissolve with methanol filters, filtrate adds organic solvent-acetone, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins according to existing patent documentation or scientific and technical literature prepare (Yang Chongren, kingdom swallow, 5 jewels, etc., the saponin component of pseudo-ginseng reed head, pharmacy circular, 1985,20 (6): 337-338; ).
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 24
Get Radix Notoginseng total arasaponins and be dissolved in water, filter, filtrate is by the HPD-100 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collect 50% elutriant, drying, dry thing dissolve with ethanol filters, filtrate adds the organic solvent ethyl acetate, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins is by commercially available purchase,
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 25
Get pseudo-ginseng and extract the Radix Notoginseng total arasaponins obtain and be dissolved in water, filter, filtrate is by the HPD-450 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collects 50% elutriant, dry, dry thing dissolve with ethanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins preparation method is prepared according to existing patent documentation or scientific and technical literature;
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 26
Get genseng and extract the Radix Ginseng total saponins obtain and be dissolved in water, filter, filtrate is by the HPD-300 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collects 50% elutriant, dry, dry thing 95% dissolve with ethanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Radix Ginseng total saponins extracting method prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 27
Get Radix Panacis Quinquefolii and extract and to obtain total saponins and be dissolved in water, filter, filtrate is by the HPD-100A macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collects 50% elutriant, dry, dry thing dissolves with propyl carbinol, filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described American ginseng total saponins extracting method prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 28
Get pseudo-ginseng and extract the total saponins obtain and be dissolved in water, filter, filtrate is by the AB-8 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collects 50% elutriant, dry, dry thing dissolve with methanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
The extracting method of described Radix Notoginseng total arasaponins can lead to the existing patent documentation of basis or scientific and technical literature prepares.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 29
Get gynostemma total saponin and be dissolved in water, filter, filtrate is by 1400 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collects 50% elutriant, dry, dry thing dissolve with ethanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described gynostemma total saponin prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 30
Get Radix Ginseng total saponins and be dissolved in water, filter, filtrate is by 1400 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collects 50% elutriant, dry, dry thing 95% dissolve with ethanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Radix Ginseng total saponins prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 31
Get pseudo-ginseng and extract and to obtain total saponins and be dissolved in water, filter, filtrate is by the D101 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collects 50% elutriant, dry, dry thing dissolves with propyl carbinol, filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins extracting method prepares according to existing document.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 32
Get genseng and extract and to obtain total saponins and be dissolved in water, filter, filtrate is by the HPD-450 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collects 50% elutriant, dry, dry thing dissolve with methanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Radix Ginseng total saponins prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 33
Get gynostemma pentaphylla and extract the total saponins obtain and be dissolved in water, filter, filtrate is by the HPD-400A macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collects 50% elutriant, dry, dry thing 95% dissolve with ethanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described gynostemma total saponin extracting method is prepared according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 34
Get Radix Ginseng total saponins and be dissolved in water, filter, filtrate is by 1400 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collects 50% elutriant, dry, dry thing 95% dissolve with ethanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Radix Ginseng total saponins prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 35
Get gynostemma pentaphylla and extract and to obtain total saponins and be dissolved in water, filter, filtrate is by the D101 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collects 50% elutriant, dry, dry thing dissolves with propyl carbinol, filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described gynostemma total saponin extracting method is prepared according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 36
Get pseudo-ginseng and extract and to obtain total saponins and be dissolved in water, filter, filtrate is by the AB-8 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collects 50% elutriant, dry, dry thing dissolve with methanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 37
Get Radix Notoginseng total arasaponins and be dissolved in water, filter, filtrate is by the 1300-I macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collects 50% elutriant, dry, dry thing dissolve with methanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 38
Get Radix Ginseng total saponins and be dissolved in water, filter, filtrate is by the D101 macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collects 50% elutriant, dry, dry thing dissolve with ethanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Radix Ginseng total saponins is by commercially available purchase,
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
The foregoing description 11-38 obtains in ginsenoside Rb1's effective constituent, ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, impurity ginsenoside Rh
1Content greater than 0 and less than 5%; Perhaps ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, impurity ginsenoside Rh
1Content more than or equal to 0.1% and less than 5%; Perhaps ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, impurity ginsenoside Rh
1Content more than or equal to 0.5% and smaller or equal to 3.0%;
The ginsenoside Rb of the foregoing description 1-38
1Be the pharmaceutical preparation of active substance preparation, wherein unitary dose is 1mg-4000mg.
The foregoing description 1-38 above-mentioned ginsenoside Rb
1Be the pharmaceutical preparation of active substance preparation, wherein unitary dose is 5mg-2000mg.
Above-mentioned ginsenoside Rb
1Treat and/or prevent application in cardiovascular and cerebrovascular diseases, tumour, aging, the hypomnesis medicine in preparation.
Annotate: the present invention's concrete technical scheme required for protection is not limited to the concrete combination of the expressed technical scheme of the foregoing description.
Claims (11)
1, a kind of ginsenoside Rb
1, ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, and foreign matter content is characterized in that greater than 0 and smaller or equal to 10% impurity comprises the ginsenoside Rh
1, the ginsenoside Rh
1Content is greater than 0 and less than 5%.
2, a kind of ginsenoside Rb according to claim 1
1, impurity ginsenoside Rh wherein
1Content more than or equal to 0.1% and less than 5%.
3, a kind of ginsenoside Rb according to claim 1
1, impurity ginsenoside Rh wherein
1Content more than or equal to 0.5% and smaller or equal to 3%.
4, according to each described a kind of ginsenoside Rb of claim 1-3
1, its preparation method is: get pseudo-ginseng, genseng, Radix Panacis Quinquefolii or gynostemma pentaphylla and extract the total saponins that obtains, total saponins is dissolved in water, filter, filtrate is passed through nonpolar or low-pole macroporous adsorptive resins, washing, 40% ethanol elution, elutriant discards, and uses 50% ethanol elution again, collects 50% elutriant, drying, dry thing filters with ethanol, 95% ethanol, propyl carbinol or dissolve with methanol, filtrate adds or does not add organic solvent, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
5, according to each described a kind of ginsenoside Rb of claim 1-3
1, its analysing and detecting method is:
Adopt high performance liquid chromatography to carry out check and analysis; Chromatographic column: C
18Reverse-phase chromatographic column; Chromatographic condition and system suitability experiment: with the octadecylsilane chemically bonded silica is weighting agent; Flow velocity 1.0ml/min; 25 ℃ of column temperatures; Detect wavelength 203nm; Number of theoretical plate is pressed ginsenoside Rb
1Meter should be not less than 2500; With water: acetonitrile is a moving phase, carries out gradient elution by following condition of gradient elution, moves 60 minutes;
In the time of 0-40 minutes, the ratio of water reduces to 50% by 80%, and the ratio of nitrile rises to 50% by 20%; In the time of 40-43 minutes, the ratio of water reduces to 20% by 50%, and the ratio of nitrile rises to 80% by 50%; In the time of 43-48 minutes, keeping the ratio of water is 20%, and keeping, the ratio of nitrile is 80%; In the time of 48-50 minutes, the water ratio rises to 80% by 20%, and the ratio of water reduces to 20% by 80%; 50-60 minutes keep the ratio of water is 20%, the ratio 80% of nitrile;
The preparation of reference substance solution: precision takes by weighing ginsenoside Rb
1Reference substance adds dissolve with methanol and shakes up in volumetric flask, and is diluted to scale;
The preparation of need testing solution: precision takes by weighing or measures sample and adds dissolve with methanol in the volumetric flask and shake up, and is diluted to scale;
Assay method: accurate respectively absorption reference substance solution and need testing solution respectively inject liquid chromatograph, adopt by external standard method with calculated by peak area, promptly.
6, according to each described a kind of ginsenoside Rb of claim 1-3
1Pharmaceutical preparation for the unitary dose of active substance preparation.
7, a kind of ginsenoside Rb according to claim 4
1, wherein nonpolar the or low-pole macroporous resin described in the preparation method is HPD-100, HPD-100A, HPD-300, HPD-400, HPD-400A, HPD-450, D101,1300-I, 1400 or AB-8.
8, a kind of ginsenoside Rb according to claim 4
1, wherein the organic solvent described in the preparation method is one or both in acetone, the ethyl acetate.
9, a kind of ginsenoside Rb according to claim 6
1Be the pharmaceutical preparation of active substance preparation, wherein unitary dose is 1mg-4000mg.
10, a kind of ginsenoside Rb according to claim 9
1Be the pharmaceutical preparation of active substance preparation, wherein unitary dose is 5mg-2000mg.
11, according to each described a kind of ginsenoside Rb of claim 1-6
1Treat and/or prevent application in cardiovascular and cerebrovascular diseases, tumour, aging, the hypomnesis medicine in preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007101219143A CN101392013A (en) | 2007-09-18 | 2007-09-18 | Ginsenoside Rb1 containing impurity ginsenoside Rh1 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007101219143A CN101392013A (en) | 2007-09-18 | 2007-09-18 | Ginsenoside Rb1 containing impurity ginsenoside Rh1 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101392013A true CN101392013A (en) | 2009-03-25 |
Family
ID=40492544
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2007101219143A Pending CN101392013A (en) | 2007-09-18 | 2007-09-18 | Ginsenoside Rb1 containing impurity ginsenoside Rh1 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101392013A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20150126463A1 (en) * | 2013-11-04 | 2015-05-07 | Hong Kong Baptist University | Use of herbal saponins to regulate gut microflora |
| US10456436B2 (en) * | 2013-11-04 | 2019-10-29 | Wen Luan Wendy Hsiao | Use of herbal saponins to regulate gut microflora |
-
2007
- 2007-09-18 CN CNA2007101219143A patent/CN101392013A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20150126463A1 (en) * | 2013-11-04 | 2015-05-07 | Hong Kong Baptist University | Use of herbal saponins to regulate gut microflora |
| US10456436B2 (en) * | 2013-11-04 | 2019-10-29 | Wen Luan Wendy Hsiao | Use of herbal saponins to regulate gut microflora |
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Application publication date: 20090325 |