CN1240709C - Herminium's triochistisaponine extract and its extraction refining method and its medical use - Google Patents
Herminium's triochistisaponine extract and its extraction refining method and its medical use Download PDFInfo
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Abstract
The present invention relates to a herminium triol saponin extract, an extraction and refining method and the medical application thereof, which belongs to the field of medicines. Herminium is used as a raw material; herminium total saponins are extracted by using a scientific extraction method; the total saponins are refined by macroporous adsorptive resin to obtain a product of herminium triol saponin extract; the content of panoxatriol saponins is 55% to 85%; the herminium triol saponin extract is especially suitable for preparing medicines for treating type II diabetes, etc.
Description
Technical field
The present invention relates to obtain Herba Herminii triol saponins extract its preparation method and contain the pharmaceutical composition of these extracts by extracting genseng pulp effective constituent.
Background technology
Herba Herminii is the genseng mellow fruit.Genseng is Araliaceae Araliaceae Panax panax plant panaxginseng C.A.Mayer, is famous and precious Chinese medicine.Have " main tonifying the five internal organs, peace spirit, decide soul, end palpitation with fear, remove perverse trend, the happy intelligence development that makes eye bright, for a long time obey to make light of one's life by commiting suicide prolong life " biological effect.It is documented that genseng has the branch of single stem, two stem and many stems, each stem of genseng that 3-4 gives birth to can be tied 10-30 fruit, and output is very big.
China northeast abounds with genseng, and China is the country that finds and utilize genseng the earliest that generally acknowledges in the world.Genseng has been widely applied to human body as preventing disease, health of body, treatment disease.In the last hundred years the chemical ingredients of genseng and different bioactive research thereof have been obtained breakthrough, and along with the continuous progressive achievement in research of science emerges in an endless stream.Scientific research finds that effective constituent contained in the Herba Herminii is almost just the same with the genseng root, even some composition is higher than its root, is the new efficient part that is worth development and utilization.
The scientific research result show Herba Herminii contain the ginsenoside Rb1 ,-Rb2 ,-Rc ,-Rd ,-Re ,-effective constituents such as Rg1, content is respectively 0.1%, 0.2%, 0.1%, 0.1%, 6.0%, 0.04%; Genseng is to the influence of artificial diabetes, and Chinese scholars is studied very early, and Bao Tiantong etc. study triol saponins hypoglycemic activity in the ginseng, shows that ginseng triol saponins has blood sugar reducing function well.Further the hypoglycemic activity to the monomer ginsenoside also has report successively, Japanese scholar in 1984 gives behind the rats by intraperitoneal injection Ginsenoside Rb2 10mg 8 hours, liver glycogen reduces to minimum value, p＜0.01, glucose 6-Phosphoric acid esterase level was increased to maximum value p＜0.01 in 8 hours in the liver simultaneously, and the activity of phosphofructokinase also has remarkable increase, peaks in 12 hours, experiment shows that the Ginsenoside Rb2 also suppresses gluconeogenesis and decomposition, can reduce the activity of 6-phosphogluconate enzyme in the mouse liver.After report in succession again, rats by intraperitoneal injection 10mg/0.5mlRb2 salt brine solution presents stronger hypoglycemic activity, uses after 12 hours, blood-sugar content is 573.8 ± 18.9mg/dl, and control group is 704.8 ± 31.6mg/dl; This seminar is studied ginsenoside Re's hypoglycemic activity, and experimental result shows: give diabetes rat abdominal injection ginsenoside Re 10mg/kg, the administration group tangible lowering blood glucose effect occurs in the 1-3 week of administration.Wang Qingtang etc. study the mechanism of genseng treatment artificial diabetes effect, and the mechanism that presses down the sugar effect of genseng may be the secretion that promotes β cell in the pancreas islet.
Summary of the invention
The invention provides a kind of Herba Herminii triol saponins extract, it is characterized by light yellow to brown, Powdered, bitter, UV spectrum (methyl alcohol) has maximum absorption band in 203 nanometers, high performance liquid chromatography shows the peak more than 6.The content of this Herba Herminii triol saponins extract accounts for the heavy 15%-20% of Herba Herminii jerky.
The invention still further relates to the method for preparing the said extracted thing, it comprises the steps:
(1) genseng pulp is added water at normal temperatures, mechanical stirring, 100 γ pm extract 2-3 time, extract 1-3 hour at every turn;
(2) solution of each time water being carried gained merges, and filters filtrate for later use;
(3) above-mentioned filtrate is crossed macroporous adsorbent resin, use the 10%-30% washing with alcohol then,, abandon or adopt washing lotion, continue, collect this time elutriant with the 60%-95% washing with alcohol near colourless;
(4) with behind this elutriant recovery ethanol, concentrate, drying gets Herba Herminii triol saponins extract.
The invention still further relates to and contain the pharmaceutical composition of Herba Herminii triol saponins extract, specifically, the present invention relates to contain the pharmaceutical composition that is used for the treatment of type ii diabetes of this extract as activeconstituents.
Product of the present invention or extract can be made into tablet, soft or glutoid capsule by suitable mode, be used to prepare the ready to use solution that adapts with its solubleness or the granulated powders and the injection liquid of liquid.The dosage of genseng of the present invention fruit triol saponins extract can be administered once or repeatedly in the scope of 30mg-400mg every day, preferred 200mg, and administration every day 2 times, suitable form of administration is an oral dosage form.
Characteristics of the present invention and beneficial effect are: in the drug regimen of treatment diabetes, effect is obvious as new medicinal efficient part for Herba Herminii triol saponins extract, and toxic side effect is little, the curative ratio height; The Herba Herminii aboundresources, preparation technology's method that the present invention adopts, simple, be fit to suitability for industrialized production, the yield height of triol saponins has broad application prospects.
The concrete scheme of extracting, making with extra care Herba Herminii triol saponins among the present invention is:
A. extract: with the sophisticated Herba Herminii of Araliaceae Panax is raw material, after removing seed, collection pulp, fruit juice are test raw material, the water that adopts the most cheap economy is as extracting solvent, because it is bigger that glassware for drinking water has been equipped with the solubleness of Herba Herminii triol saponins, boiling point is higher, and safety is non-environmental-pollution again, advantages such as easy recovery.
The mechanical stirring mode is carried out, adopted to the said extracted process at normal temperatures, and 100 γ pm accelerate to extract, water with the meat 3 times of amounts of a certain amount of Herba Herminii, extract respectively 3 times, filter united extraction liquid, concentrating under reduced pressure, 0.07Mp, to flowing paste, proportion 1.11-1.21, get the thick product of Herba Herminii total saponins, cryopreservation.
B. refining: it is a certain amount of to get Herba Herminii total saponins crude product, the water dilution of 5 times of amounts of water, macroporous adsorbent resin D101 on the diluent, at first water is eluted to colourless, use 30% ethanol elution to colourless then, use the 70%-85% ethanol elution at last, collect last elutriant, reclaim eluent and get Herba Herminii triol saponins, productive rate 10%-20%.Purity contains Herba Herminii triol saponins 55%-85%.
The advantage of extraction of the present invention, process for purification is to be raw material with the Herba Herminii, at normal temperatures and pressures, is that solvent extracts the thick product of Herba Herminii total saponins with water; And then make with extra care Herba Herminii triol saponins with macroporous adsorbent resin, and method is simple, does not need other deleterious organic solvent, and the yield height has reached the purity of drug manufacture.
Herba Herminii triol saponins has the pharmacologically active for the treatment of type ii diabetes preferably, and the diabetes traditional Chinese medical science is referred to as diabetes.Thereby Herba Herminii triol saponins can be used for preparing medicine, is specially adapted to treat diseases such as diabetes.
In order to prepare medicine with Herba Herminii triol saponins, be made into a kind of pharmaceutical preparation of form, said preparation also has the vehicle of solid or liquid except that activeconstituents.The formulation of medication and administration is normally such, and under the situation for oral administration, it can any conventionally form administration, as powder, granula, tablet, capsule, pill etc., and solution such as suspension agent, syrup etc., buccal tablets, sublingual lozenge etc.Here the vehicle of solid of Shi Yonging or liquid is being known in the art.
Herba Herminii triol saponins preparation treatment diabetes medicines such as (diabeteses) is to be made of effective constituent monomer or the effective constituent vehicle with solid or liquid.Below for several concrete examples: powder is if powder agent for oral administration, its vehicle has lactose, starch, paste essence, lime carbonate, calcium phosphate, synthetic or natural pure aluminium silicate, magnesium oxide, anhydrous alumina, Magnesium Stearate, sodium bicarbonate, dry yeast etc., and its formulation comprises electuary, tablet, capsule etc.; The vehicle of solution has water, glycerine, propylene glycol, simple syrup, ethanol, fatty oil, ethylene glycol, polyoxyethylene glycol, Sorbitol Powder, Xylitol etc.;
The dosage of active substance can change according to the mode of taking, patient's age, body weight and be in a bad way degree and other similar factors.Daily dosage portion is: oral 200mg-300mg, divide secondary to take.
With the treatment diabetes medicament of Herba Herminii triol saponins preparation, its advantage is good effect, have no side effect, cheap etc.
The present invention further specifies by following experiment.
1. Herba Herminii triol saponins is to the influence of Wistar rat zoic model with hyperglycemia
Herba Herminii triol saponins 0.52g/kg, 0.26g/kg, 0.13g/kg, be large, medium and small three dosage groups, be equivalent to 8 times, 4 times, 2 times of clinical equivalent consumption respectively, wherein big or middle dosage group can make Wistar rat zoic model with hyperglycemia glucose level descend, have obvious statistical significance, P＜0.05 is with diabetes pill group 2.0g/kg, be equivalent to 4 times of clinical equivalent consumption, have similar effect; Can make the whole blood height cut the low specific viscosity of cutting of specific viscosity, whole blood and descend, relatively have obvious statistical significance P＜0.05 with model group, P＜0.01, big or middle dosage group can make plasma viscosity obviously descend, and relatively has obvious statistical significance P＜0.05 with model group.
Herba Herminii triol saponins 0.90g/kg, 0.45g/kg, 0.09g/kg, be equivalent to 10 times, 5 times, 1 times of clinical equivalent consumption respectively, the Kunming mouse glucose tolerance curve is influenced test-results to be shown, give behind the glucose 30 minutes, the heavy dose of group of Herba Herminii triol saponins blood glucose value descends and model group relatively has notable difference P＜0.05, all the other each administration group blood glucose values are lower than model group, but not statistically significant.Gave glucose 60 minutes, each is organized blood glucose value and obviously raises, and wherein each group of Herba Herminii triol saponins and diabetes pill group blood glucose value are lower than model group, but not statistically significant, and P＞0.05 is respectively organized blood glucose value in the time of can 120 minutes and is all recovered normal.
1.1 test materials
Medicine Herba Herminii triol saponins provides lot number by preclinical medicine institute of Jilin University natural drug research department: 20000708; Diabetes pill is by Guangzhou No.1 Chinese Pharmacy Factory production, authentication code: (1994) No. 11146, and the patent No.: ZL953081990.
Animal wistar rat, male and female half and half, body weight 195g-245g is provided by laboratory animal portion of Jilin University, the certification of fitness number: the moving word of doctor 10-5110 number; Main agents and instrument
U-9889 STZ U.S. sigma chemical company produces, lot number: 89H0604
Blood sugar strip Yicheng Biological Electronic Technology Co., Ltd., Beijing produces.
JPS-III type rapid blood sugar test instrument Yicheng Biological Electronic Technology Co., Ltd., Beijing produces.
Whizzer Beijing Medical Centrifugal Machine Factory, model: LDZ5-2.
Full automatic biochemical apparatus HIT produces, model: the 7170A of Hitachi type.
1.2 test method
Test conditions and method:
After animal portion of Jilin University introduces the Wistar rat, raise in animal housing of preclinical medicine institute of Jilin University, room temperature 22-25 ℃, humidity 30%-60%, feed is formed: Semen Maydis powder 39%, soya-bean cake face 25%, wheat bran face 14%, sorghum meal 10%, bone meal 2.5%, fish meal 5%, yeast 3.5%, additive 1%.
Get the confession reagent liquid that Herba Herminii triol saponins medicinal powder is made into 5.2% (g/v), 2.6% (g/v), large, medium and small three dosage of 1.3% (g/v).It is 20% (g/v) test liquid that diabetes pill is made into concentration.
Introduce 106 of Wistar rats from animal portion of Jilin University, male and female half and half adapted to for 1 week in this chamber, and fasting be can't help water after 4 hours, weighed, surveyed blood sugar, marks.Select blood glucose value in normal range, the rat of 100 male and female half and half that state is good cooked experiment, therefrom stay 10 as the normal control group, 90 Wistar rats are pressed 0.054g/kg modeling type with U-9889 STZ, U-9889 STZ is made into 2.0% solution, matching while using with 0.05mol/L citric acid solution PH=4.5.The rat fasting be can't help water after 16 hours, once injected U-9889 STZ soup by the body weight abdominal cavity.Fasting in the 8th day be can't help water after 4 hours, surveyed blood sugar, and wherein blood glucose value is that model is set up greater than 10mmol/L.With the normal control group significant difference P＜0.001 is arranged relatively.With 54 hyperglycemia model animals, be divided into five groups at random, 10～11 every group, each group of male and female is divided equally.Be grouped into: normal control group, model group, diabetes pill group 2.0g/kg are equivalent to 4 times of clinical consumption, the heavy dose of group of Herba Herminii triol saponins 0.52g/kg, middle dosage group 0.26g/kg, small dose group 0.13g/kg, are equivalent to 8,4,2 times of clinical consumption.
Model group feedwater 10ml/kg, each administration group all gives 10ml/kg for reagent liquid.Gastric infusion about at 9 o'clock in morning every day, continuously around, survey blood sugar, body weight weekly, calculate dose once by new body weight.In administration back execution all around animal, get blood and survey routine blood test, blood parameters, hemorheology and insulin content, and get pancreas and carry out histopathologic examination.Each group gained result is represented that with average x ± standard deviation S result and model group are organized a t check, judge its significance.
1.3 test-results
1.3.1 influence to blood sugar
The heavy dose of group of the 3rd all Herba Herminii triol saponins and model group comparison blood sugar P＜0.05 that obviously descends after the administration.Big or middle dosage group of Herba Herminii triol saponins and diabetes pill group blood glucose value all reduce around, with model group relatively, have obvious statistical significance P＜0.05, see Table 1.
Table 1 Herba Herminii triol saponins is to the influence of hyperglycemia model rat blood sugar value (mmol/L) (x ± s)
| Group dosage animal (only) | Normal control group 10 | Model group 10ml/kg 8 | Diabetes pill group 2.0g/kg 9 | Heavy dose of group 0.52g/kg 10 | Middle dosage group 0.26g/kg 9 | Small dose group 0.13g/kg 8 |
| Administration last Wednesday Tuesday of Zhou Sizhou | 6.12±0.53 6.19±0.55 6.12±0.41 6.12±0.31 5.85±0.46 | 17.52±2.40 17.15±2.23 15.85±3.02 14.09±3.52 13.80±4.12 | 17.22±3.22 16.85±3.06 14.57±3.02 11.97±2.53 9.36±2.83* | 17.04±3.34 15.54±3.23 13.57±2.60 10.93±2.58* 9.07±2.78* | 17.10±2.63 15.10±2.36 14.06±2.53 11.71±2.72 9.49±1.98* | 16.37±3.32 15.72±3.10 15.12±3.23 13.05±3.26 11.81±3.63 |
Compare * P＜0.05 with model group
1.3.2 influence to blood parameters
The blood glucose value and the model group of big or middle dosage group of Herba Herminii triol saponins and diabetes pill group more all have obvious decline, has statistical significance, P＜0.05, the total protein TP and the model group of diabetes pill group, heavy dose of group relatively have obvious rising, have statistical significance, P＜0.01, the blood urea nitrogen BUN and the model group of the heavy dose of group of Herba Herminii triol saponins relatively decrease, have statistical significance, P＜0.05 sees Table 2.
Table 2 Herba Herminii triol saponins is to the influence of diabetes rat biochemical indicator (x ± s)
| Group dosage animal (only) | Normal control group 10 | Model group 10ml/kg 8 | Diabetes pill group 2.0g/kg 9 | Heavy dose of group 0.52g/kg 10 | Middle dosage group 0.26g/kg 9 | Small dose group 0.13g/kg 8 |
| ALT(U/L) AST(U/L) ALP(U/L) TP(g/L) ALB(g/L) TBIL(umol/L) BUN(mmol/L) CRE2(umol/L) GLU(mmol/L) CHO(mmol/L) | 42.7±20.2 87.1±27.3 204.3±139.0 62.3±4.1 34.5±3.5 0.37±0.23 8.1±0.8 58.5±4.7 6.07±1.82 1.3±0.2 | 101.0±32.6 157.5±49.2 532.4±156.1 61.2±3.5 30.2±2.4 0.39±0.34 11.2±4.5 82.5±29.5 13.98±4.94 1.4±0.5 | 112.6±58.8 163.8±66.2 600.1±256.9 66.7±2.5** 31.2±1.8 0.17±0.11 8.9±1.0 69.8±7.7 9.33±3.29* 1.2±0.2 | 100.9±55.2 171.9±97.5 390.0±181.9 66.4±3.3** 33.7±4.1 0.36±0.32 7.5±1.6* 64.4±4.8 8.68±2.78* 1.4±0.4 | 108.2±45.9 187.7±81.9 530.1±234.4 64.6±3.5 32.3±2.4 0.24±0.20 8.8±0.9 66.1±5.8 9.27±2.76* 1.2±0.1 | 133.5±85.4 215.4±133.0 669.4±201.9 62.9±6.1 32.9±2.0 0.45±0.43 8.8±1.6 68.3±6.7 11.58±3.82 1.2+0.1 |
Compare * P＜0.05, * * P＜0.01 with model group
1.4 experiment conclusion
Herba Herminii triol saponins has the biological activity that reduces the rat experiment diabetes.
Embodiment
Genseng fruit triol saponins extract of the present invention extracts, process for purification
Embodiment one:
A. extraction: get 1.0kg genseng pulp, be dissolved in the 3L water, at room temperature, stir, under the 100 γ pm, extract 1h, extract 3 times, filter, merge filtered solution, concentrating under reduced pressure, 0.07Mpa, to certain volume, density 1.102 gets Herba Herminii total saponins crude product;
B. refining: get rough fellow and join fruit total saponin, last macroporous adsorbent resin D101 washes with water earlier to colourless, uses 30% ethanol elution instead to colourless, uses 95% ethanol elution at last, collects elutriant, be recycled to dried, must Herba Herminii triol saponins product, productive rate 20%.
Embodiment two:
A. extraction: get 1.0kg genseng pulp, be dissolved in the 3L water, at room temperature, stir, under the 100 γ pm, extract 3h, extract 2 times, filter, merge filtered solution, concentrating under reduced pressure, 0.07Mpa, to certain volume, density 1.192 gets Herba Herminii total saponins crude product;
B. refining: get rough fellow and join fruit total saponin, last macroporous adsorbent resin D4020 washes with water earlier to colourless, uses 20% ethanol elution instead to colourless, uses 75% ethanol elution at last, collects elutriant, be recycled to dried, must Herba Herminii triol saponins product, productive rate 18%.
Embodiment three:
A. extraction: get 1.0kg genseng pulp, be dissolved in the 3L water, at room temperature, stir, under the 100 γ pm, extract 2h, extract 2 times, filter, merge filtered solution, concentrating under reduced pressure, 0.07Mpa, to certain volume, density 1.192 gets Herba Herminii triol saponins crude product;
B. refining: get rough fellow and join fruit total saponin, last macroporous adsorbent resin AB-8 washes with water earlier to colourless, uses 10% ethanol elution instead to colourless, uses 50% ethanol elution at last, collects elutriant, be recycled to dried, must Herba Herminii triol saponins product, productive rate 15%.
Preparation medicine embodiment
Embodiment one:
Herba Herminii triol saponins 225g
Starch 25g
Herba Herminii triol saponins and starch are mixed, incapsulate, the 0.25g/ grain is made 1000, pressing plate, and sterilization gets final product.
Embodiment two:
Herba Herminii triol saponins 25g
Sorbic Acid 10g
Ethanol 100ml
Get Herba Herminii triol saponins and be dissolved in the ethanol, behind the adding Sorbic Acid, be diluted to 10000ml, filter with water for pharmaceutical purposes, filling bottle, the 10ml/ bottle is made 1000 bottles, and steam sterilizing 30min gets final product.
Claims (5)
1, a kind of Herba Herminii triol saponins extract, it is characterized by light yellow to brown, Powdered, bitter, UV spectrum (methyl alcohol) has maximum absorption band in 203 nanometers, high performance liquid chromatography demonstrates the peak more than 3, is mainly ginsenoside Re, Rg1, Rg2 compound, and the content of this Herba Herminii triol saponins extract accounts for the heavy 15%-20% of Herba Herminii jerky; This Herba Herminii triol saponins obtains through the following step:
(1) it is a certain amount of to get genseng pulp, the water that adds 3 times of amounts at normal temperatures, mechanical stirring, 100 γ pm extract 2-3 time, extract 1-3 hour at every turn;
(2) solution with each time poach gained merges, and filters filtrate for later use;
(3) above-mentioned filtrate is crossed macroporous adsorbent resin, use the 10%-30% washing with alcohol then,, abandon or adopt washing lotion, continue, collect this time elutriant with the 50%-95% ethanol elution near colourless;
(4) with behind this elutriant recovery ethanol, concentrate, drying gets Herba Herminii triol saponins extract.
2, a kind of extraction, process for purification of Herba Herminii triol saponins is characterized in that comprising the steps:
(1) it is a certain amount of to get genseng pulp, the water that adds 3 times of amounts at normal temperatures, mechanical stirring, 100 γ pm extract 2-3 time, extract 1-3 hour at every turn;
(2) solution with each time poach gained merges, and filters filtrate for later use;
(3) above-mentioned filtrate is crossed macroporous adsorbent resin, use the 10%-30% washing with alcohol then,, abandon or adopt washing lotion, continue, collect this time elutriant with the 50%-95% ethanol elution near colourless;
(4) with behind this elutriant recovery ethanol, concentrate, drying gets Herba Herminii triol saponins extract.
3, Herba Herminii triol saponins preparation method of extract according to claim 2, it is characterized in that: macroporous adsorbent resin is selected from D
101, D
4020, DA
201, AB-8, XAD-4 or D
H
4, a kind of pharmaceutical composition contains the described Herba Herminii triol of claim 1 saponins extract as activeconstituents.
5, the application of Herba Herminii triol saponins extract as claimed in claim 1 in the medicine of preparation treatment type ii diabetes.
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| CN101948497B (en) * | 2010-09-03 | 2012-08-15 | 吉林圣亚医药科技有限公司 | Ginsenoside R10, extraction method and pharmaceutical use thereof |
| CN102796159B (en) * | 2011-05-24 | 2014-12-03 | 复旦大学 | Dammarane glucosides and preparation method and application thereof |
| CN103549313B (en) * | 2013-11-01 | 2015-07-22 | 王颖 | Method for extracting ginseng fruit essence materials |
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