CN1900100A - Rehmannia root oligose and its extracting method - Google Patents
Rehmannia root oligose and its extracting method Download PDFInfo
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- CN1900100A CN1900100A CN 200610088850 CN200610088850A CN1900100A CN 1900100 A CN1900100 A CN 1900100A CN 200610088850 CN200610088850 CN 200610088850 CN 200610088850 A CN200610088850 A CN 200610088850A CN 1900100 A CN1900100 A CN 1900100A
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Abstract
The present invention discloses a kind of rehmannia root oligose and its extraction process. The extraction process includes following steps: crushing rehmannia root; soaking rehmannia root in water for 1-10 hr; collecting the supernatant; decompression concentrating the supernatant to obtain concentrate or volatilizing the supernatant at 40-60 deg.c to obtain extractum; dissolving the concentrate or the extractum in 70-100 % concentration alcohol; filtering to obtain filtrate; vacuum concentrating at 20-60 deg.c; ultrafiltering the concentrate to obtain ultrafiltrate with matter of molecular weight greater than 5 kD eliminated; and adsorbing the ultrafiltrate with active carbon to eliminate pigment and obtain rehmannia root oligose.
Description
Technical field
The present invention relates to compound and extracting method thereof, particularly relate to rehmannia root oligose and extracting method thereof.
Background technology
Cerebrovascular disease is a kind of common disease of serious harm people ' s health, and its sickness rate, morbidity, mortality ratio and disability rate are all very high.Statistical information shows that the elderly of middle national trouble cardiovascular and cerebrovascular diseases reached 1.3 hundred million in 2000, accounted for 11% of China's population; According to a preliminary estimate, the gerontal patient of this class disease will reach 2.3 hundred million to the year two thousand fifty, account for 20% of China's population.Wherein, the sickness rate of over-65s elderly brain vascular disease and brain dementia is more than 15%.Cerebrovascular disease and brain dementia not only can cause the elderly's dyskinesia, also can cause the obstacle of the elderly's language and spiritual aspect, thereby reduce its quality of life, therefore, press for the health care medicine of enhancing the elderly physique and the medicine of the elderly's nervous system disorders.
Glutinous rehmannia is the scrophulariaceae per nnial herb, its fresh or dried root, the beginning be stated from Shennong's Herbal, classify as top grade, sweet, the tepor of nature and flavor.Return liver, kidney channel, have the enriching yin of enriching blood, beneficial essence is filled out the marrow effect.
Modern pharmacology proves, that the effective constituent rehmannia root oligose in the glutinous rehmannia has is anti-ageing, intelligence development, enhancing immunity is regulated and effect such as treatment cardiovascular and cerebrovascular diseases, therefore, presses for a kind of extracting method of simple and economic and practical rehmannia root oligose.
Summary of the invention
The extracting method that the objective of the invention is a kind of simple and economic and practical rehmannia root oligose.
The extracting method of rehmannia root oligose provided by the present invention may further comprise the steps:
1) pulverizes glutinous rehmannia;
2) glutinous rehmannia is soaked in water 1-10 hour;
3) throw out in the removal leach liquor is collected leach liquor;
4) to the supernatant liquor concentrating under reduced pressure, reclaim concentrated solution, or under 40-60 ℃, supernatant liquor is volatilized, obtain medicinal extract;
5) be 1 with concentrated solution and volumn concentration (V/V) by volume for the ethanol of 70-100%: the 3-7 mixing, or medicinal extract is dissolved in the 70-100% ethanol, with the precipitation polysaccharide;
6) mixed solution that step 5) is obtained filters, and collects filtered liquid;
7) filtered liquid is concentrated at 20-60 ℃ of following vacuum decompression, remove ethanol simultaneously, reclaim concentrated solution;
8) concentrated solution is carried out ultrafiltration, remove molecular weight, collect ultrafiltrated greater than 5000 daltonian materials;
9) ultrafiltrated is adsorbed with depigmentation with gac, obtain the rehmannia root oligose extracting solution.
In the said extracted method, for obtaining better extraction effect, preferably glutinous rehmannia is crushed to the particle of particle diameter in the step 1) less than 2mm, extract again.
Step 2) water that is used for lixiviate in is preferably water for injection.
Available centrifugation method is removed the throw out in the leach liquor in the step 3).
Be to improve raw material availability, can be with this step 2)-3) the merging leach liquor repeated 1-3 time.
Concentrating under reduced pressure in the step 4) can carry out under 10-60 ℃, is preferably 20-50 ℃.
Usage aperture is that the filter membrane of 0.4-0.5mm filters mixed solution in the step 6), and described filter membrane aperture is preferably 0.45mm.
Usage aperture is that the ultra-filtration membrane of 10-0.2 μ m or the scope of holding back are that the ultra-fine filter of 3000-20000 carries out ultrafiltration to concentrated solution in the step 8).
The said extracted method all is suitable for Rehmannia Root and Radix Rehmanniae.
In addition, the rehmannia root oligose that obtains with aforesaid method also belongs to protection scope of the present invention.
Simultaneously, the rehmannia root oligose that can obtain with aforesaid method is the curative drug that activeconstituents is prepared into cardiovascular and cerebrovascular diseases and sequela, diabetes and constipation and other diseases.
The content of bulk drug rehmannia root oligose should be greater than 85% in the said medicine.
When needing, in application of the present invention, can also add one or more acceptable accessories, described auxiliary material comprises thinner, vehicle, weighting agent, tackiness agent, wetting agent, absorption enhancer, tensio-active agent, lubricant, stablizer of pharmaceutical field routine etc., also can add flavouring agent, sweeting agent and pigment etc. in case of necessity.
Application of the present invention can also be made multiple medicament forms such as tablet, granula, pulvis, oral preparation, injection liquid except that making capsule.The medicine of above-mentioned various formulations all can be according to the ordinary method preparation of pharmaceutical field.
The oral consumption of the adult of said medicine is generally 400mg/kg/d, can use by one or many, and be 10 to 20 days the course of treatment.
The invention provides a kind of extracting method of effective rehmannia root oligose.This extracting method has the following advantages: 1, simple to operate, be easy to carry out industrial applications; 2, extraction cost is cheap; 3, the toxicity of used extraction solvent is less, and extracting cycle is short; 4, rehmannia root oligose content height in the extracting solution, content can reach more than 85%, and molecular weight is less than 5000 dalton.Simultaneously, the rehmannia root oligose that can obtain with the inventive method is the curative drug that activeconstituents is prepared into cardiovascular and cerebrovascular diseases and sequela, diabetes and constipation and other diseases.The present invention lays a good foundation for the scale operation of rehmannia root oligose, and will play a great role in effective utilization of rehmannia root oligose, having a extensive future of field of medicaments.
Below in conjunction with specific embodiment the present invention is described in further details.
Description of drawings
Fig. 1 is the schema of rehmannia root oligose extraction process of the present invention
Embodiment
Method therefor is ordinary method if no special instructions among the following embodiment.
The extraction of embodiment 1, rehmannia root oligose and quality evalution thereof
One, the extraction of rehmannia root oligose
Selection meet version in 2005 " the Radix Rehmanniae medicinal material of Chinese pharmacopoeia standard therefrom extracts rehmannia root oligose with following method, and the schema of extraction process is seen Fig. 1, may further comprise the steps:
1) get it filled 20 kilograms of materials clean up, and pulverizing medicinal materials is become particle less than 2mm;
2) with 50 liters of waters for injection the glutinous rehmannia particle was soaked 5 hours;
3) leach liquor is carried out centrifugal, obtain about 45 liters of leach liquor;
4) under 35 ℃ with supernatant liquor drying under reduced pressure to 20 liter;
5) with concentrated solution and 80 liters of volumn concentrations be 95% ethanol mixing;
6) mixed solution that step 5) is obtained is that the filter membrane of 0.45mm filters with the aperture, obtains about 100 liters of filtered liquid;
7) filtered liquid is concentrated into 20 liters at 35 ℃ of following vacuum decompressions;
8) be 5000 daltonian ultra-fine filters (available from film device limited liability company of Beijing rising sun nation) ultrafiltration with the concentrated solution molecular weight, obtain about 20 liters of ultrafiltrated;
9) ultrafiltrated is adsorbed with medical activated carbon, obtain about 20 liters of rehmannia root oligose extracting solution.
Two, the quality evalution of rehmannia root oligose
The yellow oligosaccharides extracting solution that step 1 is obtained carries out quality evalution from following several respects:
1, proterties
Flaxen clear liquid.
2, differentiate
(1) gets rehmannia root oligose extracting solution 1mL, add 5 of 5% α-Nai phenol ethanolic soln, shake up, slowly add vitriol oil 0.5mL, purplish red colour circle is arranged, conform to expected results at test liquid and the visible two liquid interfaces of sulfuric acid intersection along tube wall.
(2) get rehmannia root oligose extracting solution 2mL, add 1 of ninhydrin solution, shake up, heated and boiled 1 minute, putting cold back solution does not have bluish voilet, conforms to expected results.
3, high performance liquid chromatography detects high molecular weight material content
Adopt the high molecular weight material content in high performance liquid chromatography (two appendix VD of Chinese Pharmacopoeia version in 2005) the mensuration rehmannia root oligose extracting solution, with TSK-G2000swxl gel chromatographic columns (Japanese TOSOHAAS company), with acetonitrile-trifluoracetic acid-water (40: 0.1: 60) is moving phase, flow velocity is per minute 0.8mL, and the detection wavelength is 190nm.Theoretical plate number is calculated by insulin spikes should be not less than 800.Detection method is: get the rehmannia root oligose extracting solution, add water by labelled amount and make the solution that contains the 40mg rehmannia root oligose among every 1mL, get 20 μ l and inject liquid chromatograph, the record color atlas, to be considered as polymer substance prior to the peak of insulin spikes retention time, calculate high molecular weight material content by area normalization method.High molecular weight material content is less than 2% as a result, conformance with standard.
4, heavy metal content detects
According to " two appendix IXE first methods of Chinese pharmacopoeia version in 2005 detect the heavy metal content in the rehmannia root oligose extracting solution.Heavy metal content is lower than 10/1000000ths as a result, conformance with standard.
5, total sugar content is measured
1) weighing dry weight
Take by weighing about 1.5 grams of the extra large sand of calcination, pave bottom, put in the phosphorus pentoxide desiccator, under (53 ± 2) ℃, be dried to constant weight in weighing bottle.Precision takes by weighing rehmannia root oligose extracting solution 2mL, places above-mentioned weighing bottle, and drying under reduced pressure is to constant weight under uniform temp, and accurate the title decides, and measures total sugar content according to the phenol sulfuric acid process, and calculates total sugar content by following formula:
Total sugar content=total reducing sugar/dry weight * 100%
Total sugar content shows with method of the present invention to have obtained the higher rehmannia root oligose extracting solution of purity greater than more than 85% of gross dry weight as a result, and the content of rehmannia root oligose reaches 86%.
The extraction of embodiment 2, rehmannia root oligose
Selection meet version in 2005 " Rehmannia Root of Chinese pharmacopoeia standard is therefrom extracted rehmannia root oligose with following method, may further comprise the steps:
1) get it filled 20 kilograms of materials clean up, and pulverizing medicinal materials is become particle less than 2mm;
2) with 50 liters of waters for injection the glutinous rehmannia particle was soaked 8 hours;
3) leach liquor is carried out centrifugal, obtain about 45 liters of supernatant liquor;
4) under 50 ℃, supernatant liquor is volatilized, obtain medicinal extract;
5) medicinal extract is pulverized, with 100 liters of volumn concentrations be 75% ethanol mixing;
6) mixed solution that step 5) is obtained is that the filter membrane of 0.45mm filters with the aperture, obtains about 100 liters of filtered liquid;
7) filtered liquid is concentrated into 20 liters at 40 ℃ of following vacuum decompressions;
8) be 5000 daltonian ultra-fine filters (available from film device limited liability company of Beijing rising sun nation) ultrafiltration with the concentrated solution molecular weight, obtain about 20 liters of ultrafiltrated;
9) ultrafiltrated is adsorbed with medical activated carbon, obtain about 20 liters of rehmannia root oligose extracting solution.
With the method identical with embodiment 1 rehmannia root oligose that extracts is carried out quality evalution, the equal conformance with standard of the every index of result has obtained the higher rehmannia root oligose extracting solution of purity, and the content of rehmannia root oligose reaches 90%.
The extraction of embodiment 3, rehmannia root oligose and quality evalution thereof
Selection meet version in 2005 " the Radix Rehmanniae medicinal material of Chinese pharmacopoeia standard therefrom extracts rehmannia root oligose with following method, may further comprise the steps:
1) get it filled 20 kilograms of materials clean up, and pulverizing medicinal materials is become particle less than 2mm;
2) with 50 liters of waters for injection the glutinous rehmannia particle was soaked 1.5 hours;
3) leach liquor is carried out centrifugal, collect supernatant liquor;
4) repeating step 2)-step 3) 3 times, merge supernatant liquor, obtain about 55 liters of leach liquor altogether;
5) with supernatant liquor drying under reduced pressure to 20 liter;
6) with concentrated solution and 80 liters of volumn concentrations be 95% ethanol mixing;
7) mixed solution that step 5) is obtained is that the filter membrane of 0.45mm filters with the aperture, obtains about 100 liters of filtered liquid;
8) filtered liquid is concentrated into 20 liters at 30 ℃ of following vacuum decompressions;
9) be 5000 daltonian ultra-fine filters (available from film device limited liability company of Beijing rising sun nation) ultrafiltration with the concentrated solution molecular weight, obtain about 20 liters of ultrafiltrated;
10) ultrafiltrated is adsorbed with medical activated carbon, obtain about 20 liters of rehmannia root oligose extracting solution.
With the method identical with embodiment 1 rehmannia root oligose that extracts is carried out quality evalution, the equal conformance with standard of the every index of result has obtained the higher rehmannia root oligose extracting solution of purity, and the content of rehmannia root oligose reaches 88%.
Embodiment 4, experimentation on animals
1, vascular dementia test
Give the rehmannia root oligose 5 days that kunming mice abdominal injection embodiment 1 extracts, each 5mg/10g mouse is heavy, once a day, the administration mouse is carried out the vascular dementia test.The result shows diving tower and the water maze school grade and the sham operated rats no significant difference that can improve mouse than administration; Obviously reduce neuronic dead quantity.
2, mouse hypoxia endurance test
It is heavy to give the rehmannia root oligose 5mg/10g mouse that kunming mice abdominal injection embodiment 2 extracts, and is contrast with administration mouse not, and two groups of mouse are carried out hypoxia endurance test.The ability of mouse hypoxia tolerance is improved significantly after the administration as a result, and compares with control group, and the water ratio of administration group mouse hippocampal tissue reduces.
3, the hypoxia test of hippocampal neurons
Detect the damage of anoxic to hippocampal neurons, one group is the administration group: give the rehmannia root oligose 1% (mg/V) that embodiment 2 extracts, one group is the control group of not administration.The result shows that rehmannia root oligose of the present invention can reduce the damage of anoxic to hippocampal neurons, and the LDH amount of exposing obviously reduces than control group.
4, to the influence of mouse intestines
Giving kunming mice, to irritate the rehmannia root oligose 5mg/10g mouse that stomach embodiment 3 extracts heavy, and continuous 5 days once a day, is contrast with administration mouse not, observes the intestinal peristalsis frequency of two groups of mouse, and statistics intestinal beneficial bacterium quantity.The result compares with control group mice, and intestinal peristalsis frequency and the intestinal beneficial bacterium quantity of administration mouse are significantly improved.
5, the alloxan diabetes rat model is influenced
The glutinous rehmannia oligopeptides 5mg/10g that alloxan diabetes rat model abdominal injection every day embodiment 1 is extracted, continuous 7 days is contrast with the alloxan diabetes rat model of not administration, and two groups of rats are observed.The result compares with control rats, and the blood glucose value of administration rat obviously reduces.
Claims (10)
1, a kind of extracting method of rehmannia root oligose may further comprise the steps:
1) pulverizes glutinous rehmannia;
2) glutinous rehmannia is soaked in water 1-10 hour;
3) throw out in the removal leach liquor is collected leach liquor;
4) to the supernatant liquor concentrating under reduced pressure, reclaim concentrated solution, or under 40-60 ℃, supernatant liquor is volatilized, obtain medicinal extract;
5) be that the ethanol of 70-100% is 1 by volume with concentrated solution and volumn concentration: the 3-7 mixing, or medicinal extract is dissolved in the 70-100% ethanol;
6) mixed solution that step 5) is obtained filters, and collects filtered liquid;
7) filtered liquid is concentrated at 20-60 ℃ of following vacuum decompression, reclaim concentrated solution;
8) concentrated solution is carried out ultrafiltration, remove molecular weight, collect ultrafiltrated greater than 5000 daltonian materials;
9) ultrafiltrated is adsorbed with depigmentation with gac, obtain the rehmannia root oligose extracting solution.
2, extracting method according to claim 1 is characterized in that: the glutinous rehmannia in the described step 1) comprises Rehmannia Root and Radix Rehmanniae; Glutinous rehmannia is crushed to the particle of particle diameter less than 2mm.
3, extracting method according to claim 1 is characterized in that: the water that is used for lixiviate described step 2) is water for injection.
4, extracting method according to claim 1 is characterized in that: remove throw out in the leach liquor with centrifugal or filtering method in the described step 3).
5, extracting method according to claim 1 is characterized in that: with described step 2)-step 3) repeats the merging leach liquor 1-3 time.
6, extracting method according to claim 1 is characterized in that: the concentrating under reduced pressure in the described step 4) carries out under 10-60 ℃.
7, extracting method according to claim 1 is characterized in that: be that the filter membrane of 0.4-0.5mm filters mixed solution with the aperture in the described step 6).
8, extracting method according to claim 1 is characterized in that: be that the ultra-filtration membrane of 10-0.2 μ m or the scope of holding back are that the ultra-fine filter of 3000-20000 carries out ultrafiltration to concentrated solution with the aperture in the described step 8).
9, the rehmannia root oligose that obtains with each described method of claim 1-8.
10, the application of the described rehmannia root oligose of claim 9 in preparation cardiovascular and cerebrovascular diseases and sequela, diabetes and constipation therapy medicine.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112870271A (en) * | 2021-02-08 | 2021-06-01 | 河南农业大学 | Preparation method of high molecular weight rehmannia root polysaccharide and method for obtaining prepared rehmannia root by high-pressure steaming of fresh rehmannia root |
| CN113476525A (en) * | 2021-08-11 | 2021-10-08 | 江苏省中医药研究院 | Traditional Chinese medicine glucoside composition with lipid-lowering and weight-losing effects and preparation method and application thereof |
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| CN1157399C (en) * | 2000-08-25 | 2004-07-14 | 陈梅英 | Extracting method for stachyose |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112870271A (en) * | 2021-02-08 | 2021-06-01 | 河南农业大学 | Preparation method of high molecular weight rehmannia root polysaccharide and method for obtaining prepared rehmannia root by high-pressure steaming of fresh rehmannia root |
| CN112870271B (en) * | 2021-02-08 | 2022-04-19 | 河南农业大学 | Preparation method of high molecular weight rehmannia root polysaccharide and method for obtaining prepared rehmannia root by high-pressure steaming of fresh rehmannia root |
| CN113476525A (en) * | 2021-08-11 | 2021-10-08 | 江苏省中医药研究院 | Traditional Chinese medicine glucoside composition with lipid-lowering and weight-losing effects and preparation method and application thereof |
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