CN1047088C - Process for extracting active ingredients from changium root and medicinal use - Google Patents
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Abstract
The present invention provides a method for extracting active ingredients from changium root, which comprises two sets of extraction processes of aqueous extraction and ethanol extraction. Simultaneously, the present invention provides a method for directly preparing a proper preparation from the active ingredients of the changium root or preparing the proper preparation by the way that the active ingredients of the changium root is mixed with solids or liquid usually used for the preparation in pharmacy. The present invention also introduces an application of the active ingredient preparation of the changium root for treating some diseases.
Description
The invention belongs to Chinese medicine development field, relate to the extracting method of active ingredients from changium root and the production and processing method of active ingredients from changium root preparation.
Radix Changii (Changium smyrnioides wolff) is the distinctive Umbelliferae of China (Umbelliferae) autogenus plant, its root hyoscine, in the herbal books such as bright clear grade record its cure mainly " miasma swells and ache, generates heat ", " cough dyspnea, expectorant stop up that fire rises disease of a specified duration, drenches drop, difficult labour amenorrhea dysentery; ... lung-heat regurgitation, caye diaphragm ... wait disease ".Though the medicinal history in existing three, 400 years of Radix Changii, the extraction of its effective ingredient and utilize its effective ingredient production and processing to become the unprecedented report of preparation.
The object of the present invention is to provide the extracting method of active ingredients from changium root, 〉=and utilize its several preparations of effective ingredient production and processing, with its generation, adjusting immunologic function, mutation as the lipid peroxide of antithrombotic, blood fat reducing, inhibition hypercholesterolemia, improve stress ability, improve the medicine of intestinal function.
Task of the present invention is finished in the following manner, and the extracting method of described active ingredients from changium root in turn includes the following steps:
(1) at first the Radix Changii root is put and boiled to the no white heart in the boiling water, take out and to scrape off crust, be cut into decoction pieces after drying; It is characterized in that:
(2) get that above-mentioned decoction pieces adds water that 10-20 doubly measures, puts in the tilting steam heated pot, heating 1-2 hour, filtered while hot, residue add 10-20 times of water gaging heating 1-2 hour again, filtered while hot, merging filtrate leave standstill, and draw supernatant and get the water extract;
(2,1) with above-mentioned water extract put again in the steam heated pot, worn-out mouthful concentrate, drying under reduced pressure then, Radix Changii decocting thing; Or
(2,2) get that above-mentioned water extract adds 2 times of amount ethanol, leaves standstill filtrations, filtrate elder generation concentrating under reduced pressure, drying under reduced pressure again, must the Radix Changii water extract; Perhaps
(3) get by technical process (1) gained Radix Changii decoction pieces, add 10 times of amount 70%-95% ethanol, placed the multi-function extractor hot reflux 1-2 hour, filtered while hot, add 10 times of amount 70%-95% alcohol heat reflux 0.5-1.5 hour again, filtered while hot, residue, simultaneously merging filtrate or get by technical process (1) gained Radix Changii decoction pieces, add the 70%-95% ethanol percolation colourless to solution, residue and percolate;
(4) with above-mentioned filtrate or percolate decompression recycling ethanol, gained extractum gets the Radix Changii ethanol extract after drying; Or
(5) will be by the residue obtained drying of technical process (3)
(6) exsiccant residue adding 5-10 is doubly measured upright heat and carry 1-2 hour, filtered while hot, residue adds 5-10 times of water gaging heat again and carries 1-2 hour, filtered while hot, merging filtrate
(7) above-mentioned filtrate suitably concentrated, leave standstill, the filtering insoluble matter, add 3 times of amount ethanol precipitations then, the leaching precipitation, vacuum drying gets the Radix Changii crude polysaccharides.
The extracting method of above-mentioned active ingredients from changium root can find out clearly that in fact it is the difference according to required effective component extracting, and adopted with water as extractant and with ethanol as this two covers extraction process process of extractant.
Therefore, the extracting method of above-mentioned active ingredients from changium root also is appreciated that into following extraction step:
(1) at first the Radix Changii root is put boiling water and boiled, take out and to scrape off crust, dry and become decoction pieces later age to the no white heart;
(2) get above-mentioned decoction pieces adding 10-20 and doubly measure 0%-95% ethanol, heat was extracted 1-2 hour, filtered while hot, and residue adds 10-20 again and doubly measures 0%-95% ethanol, and heat was extracted 0.5-1.5 hour, got residue, simultaneously merging filtrate.Or get by technical process (1) gained Radix Changii decoction pieces, it is colourless to solution to add the 0%-95% ethanol percolation, gets residue and percolate.
(3) get above-mentioned 0% ethanol extract (being the water extract) and put in the steam heated pot, uncovered concentrating, drying under reduced pressure gets Radix Changii decocting thing then; Or
(4) get above-mentioned 0% ethanol extract (being the water extract) and add 2 times of amount ethanol, leave standstill filtration, filtrate elder generation concentrating under reduced pressure, drying under reduced pressure gets the Radix Changii water extract again; Or
(5) get above-mentioned 0% ethanol extract (being the water extract) and suitably concentrate, leave standstill, the filtering insoluble matter adds 3 times of amount ethanol precipitations then, the leaching precipitation, and vacuum drying gets the Radix Changii crude polysaccharides; Or
(6) get above-mentioned 10-95% ethanol extract decompression recycling ethanol, gained extractum drying under reduced pressure gets the Radix Changii ethanol extract.
Be described further below by the extracting method of specific embodiment active ingredients from changium root provided by the present invention.
[embodiment 1]
(1) pre-treatment: get the Radix Changii root and put and boil in the boiling water, take out, scrape off crust and dry, be cut into decoction pieces to the no white heart.
(2) get by technical process (1) gained Radix Changii decoction pieces, add 10 times of water, put in the tilting steam heated steel, heated 1.5 hours, filtered while hot, residue adds 10 times of water again, heated 1.5 hours, filtered while hot, filtrate merges, and leaves standstill, and draws supernatant and gets the water extract.
(3) the water extract is put in the steam heated pot again, after worn-out mouth concentrated, 70 ℃ of drying under reduced pressure got Radix Changii decocting thing.
[embodiment 2]
(1) pre-treatment: identical with the technical process (1) of example 1
(2) heat is extracted: identical with the technical process (2) of example 1
(3) alcohol precipitation: get by technical process (2) gained water extract add 2 times of amount ethanol, leave standstill, after filtration, filtrate alkali press and concentrate, 70 ℃ of drying under reduced pressure, the Radix Changii water extract.
[embodiment 3]
(1) pre-treatment: identical with the technical process (1) of example 2
(2) hot reflux or percolation: get by technical process (1) gained Radix Changii decoction pieces 10Kg, add 10 times of amount 70%-95% ethanol, hot reflux is 1.5 hours in multi-function extractor, filtered while hot, add 10 times of amount 70%～95% alcohol heat reflux 1 hour again, filter merging filtrate.Or taking technique process (1) gained Radix Changii decoction pieces 10Kg, it is colourless to solution to add 70%～95% ethanol percolation, gets percolate.
(3) reclaim ethanol: with filtrate or percolate decompression recycling ethanol, 70 ℃ of dryings of gained extractum get the Radix Changii ethanol extract.
[embodiment 4]
(1) pre-treatment: identical with the technical process (1) of example 2
(2) defat: get by technical process (1) gained Radix Changii decoction pieces 10Kg, extracted 6 hours the residue drying with industrial reflow of alcohol.
(3) get by technical process (2) is residue obtained and add 10 times of water gaging heat and carry 1.5 hours, filtered while hot, residue adds 10 times of water gaging heat again and carries 1.5 hours, filtered while hot, merging filtrate.
(4) precipitate with ethanol: get by technical process (3) gained water extract and suitably concentrate, leave standstill, filtering insoluble matter, supernatant add 3 times of amount ethanol precipitations, the leaching precipitation, vacuum drying, canescence Radix Changii crude polysaccharides.
Utilize the physicochemical property of the active ingredients from changium root that method provided by the invention obtains as follows:
1, Radix Changii decocting thing and water extract: the sample thief thing adds that water is suitable put dissolving after, (1) gets the 1ml sample liquid, add 0.2% ninhydrin solution, heating is 5 minutes on boiling water is molten, indigo plant or bluish violet appear after cold, or with sample liquid point on filter paper, spray 0.2% ninhydrin solution after, baking is 2 minutes about 100 ℃, presents the aubergine speckle.(2) get the 1ml sample liquid; Add 5% alpha-Naphthol test solution, shake up, drip about 1ml concentrated sulphuric acid along tube wall, two liquid produce purplish red colour circle at the interface, (3) an amount of standardize solution of sample thief liquid, accurate absorption sample liquid and asparagine titer are put on same block of efficient lamellae, launch 7cm with n-butyl alcohol-glacial acetic acid-alcohol-water (15: 4.5: 15: 5), taking-up volatilizes, spray 0.2% 1,2,3-indantrione monohydrate liquid, 100 ℃ of bakings 2 minutes, place 1 hour in the dark place after, put quantitatively (condition determination: λ of CS-930 thin-layer chromatogram scanner scanning, 370nm, reflective zigzag scanning, SX=3, slit 1.2 * 1.2mm) is obtained the percentage composition of sample Mid-Heaven Gate winter amide by standard curve.The content of Radix Changii Mid-Heaven Gate winter amide should be not less than 0.7%.
2, Radix Changii ethanol extract: after the sample thief thing added the suitable heavily dissolving of ethanol, (1) got the 1ml sample liquid, adds 5% alpha-Naphthol test solution, shakes up, and drips about 1ml concentrated sulphuric acid along tube wall, and two liquid produce purplish red Bao Huan at the interface.(2) get the 1ml sample liquid, add oxammonium hydrochloride. test solution and 10% potassium hydroxide ethanol liquid to being alkalescence, be heated in water-bath and react completely, cooling adds hcl acidifying, adds 1% ferric chloride test solution again, solution is red, purple.(3) get the 1ml sample liquid, add equal-volume 3% sodium carbonate test solution, in water-bath, boiled 3-5 minute, put coldly, add diazo reagent, show red, purple.
3, Radix Changii crude polysaccharides: after sample thief liquid added the suitable quantity of water dissolving, (1) got the 1ml sample liquid, and the fehling reagent 4-5 that adds new preparation drips, and heating is several minutes on boiling water bath, produces red-brown precipitation.(2) sample thief thing 20mg, with 1mol/l sulphuric acid tube sealing hydrolysis (100 ℃) 6 hours, saturated barium hydroxide water liquid neutralization, elimination post precipitation, concentrating under reduced pressure.Concentrated solution is put on Xinhua's chromatography filter paper at a slow speed (long 40cm), with monosaccharide standard substance liquid on the time point, with ethyl acetate-pyridine-water (10: 4: 3) is developing solvent, aniline-phthalic acid test solution colour developing, with rhamnose, arabinose, xylose, mannose, galactose, these six kinds of corresponding positions of standard substance of glucose on show brown speckle.
On the basis of the active ingredients from changium root that obtains with said method, its production and processing can be become generation as the lipid peroxide of antithrombotic, blood fat reducing, inhibition hypercholesterolemia, regulate immunologic function, mutation, raising stress ability, improve the medicine of intestinal function, active ingredients from changium root as active constituents of medicine, directly or with solid that pharmaceutically is generally used for such preparation or liquid is made suitable dosage form.
Below we continue by embodiment the preparation method of active ingredients from changium root preparation to be described further:
[embodiment 5]
Get Radix Changii decocting thing (or water extract, ethanol extract) dry, pulverize, cross 80 mesh sieves, directly or add suitable diluents (Radix Changii medical material dry powder, dextrin, microcrystalline cellulose etc.) fill capsule
[embodiment 6]
Get Radix Changii crude polysaccharides (or decocting thing, water extract, ethanol extract) lyophilization, pulverizing, cross 40 mesh sieves, the agent of lyophilizing flour bag is made in fill
[embodiment 7]
Get Radix Changii ethanol extract (or decocting thing, water extract, crude polysaccharides) by 1: 1～1: 4 diluent (dextrin, starch etc.), dry, pulverize, granulate, tabletting or make pill.
[embodiment 8]
Get Radix Changii ethanol extract (or decocting thing, water extract, crude polysaccharides) and add the suitable quantity of water dissolving, filter, add simple syrup, sodium benzoate, cold preservation filters, and is packaged in the 10ml ampoule, and 100 ℃ of sterilization 30min are promptly.
[embodiment 9]
Get Radix Changii water extract (or decocting thing, ethanol extract, crude polysaccharides) and add the suitable quantity of water dissolving, filter, add Radix Changii volatile oil, simple syrup, sodium benzoate makes oral liquid with example 8 technologies.
[embodiment 10]
Get Radix Changii ethanol extract (or decocting thing, water extract, crude polysaccharides) and add the suitable quantity of water dissolving, go tannin, filter, fill, sterilization with gelatin or polyamide.
[embodiment 11]
Get Radix Changii volatile oil and gelatin solution and make by the drop pill machine and biphasely ooze by friction speed, make a certain amount of gelatin solution with quantitative Radix Changii volatile oil parcel after, cooled and solidified becomes soft gelatin capsule.
[embodiment 12]
Get Radix Changii ethanol extract (or decocting thing, water extract, crude polysaccharides) and add an amount of diluent, make microcapsule with spray drying method.
Active ingredients from changium root and preparation thereof with the said method gained can be used as antithrombotic, fall the blood ester, suppress the generation of the lipid oxidation thing of hypercholesterolemia, regulate immunologic function, mutation, raising stress ability, improve the medicine of intestinal function.Medication is oral or injection, dosage 1-6 gram every day (with active ingredients from changium root, i.e. decocting thing, water extract, ethanol extract or crude polysaccharides meter), and 1 month is a course of treatment.
Illustrate its drug action with pharmacological tests below:
1, studies have shown that to the regulating action of immunologic function is existing the Chinese medicine benefiting action is relevant with raise immunity.This experimentation shows that Radix Changii has multiple pharmacologically active to improving body's immunity.Radix Changii and polysaccharide thereof demonstrate significant immunologic enhancement on the one hand: 1: 10 Radix Changii decocting liquid of (1) lumbar injection can obviously improve the NK activity of mouse boosting cell, relatively improves 60.81%.P＜0.01 with matched group.Illustrate that Radix Changii has facilitation to mice NK activity, but enhancing body gives protection against cancer, the anti-infective ability.(2) oral administration gavage Radix Changii decocting liquid (15g/Kg) and polysaccharide (30mg/Kg) can significantly improve normal mouse peritoneal macrophage YC-garland formation rate, matched group is 42.414 ± 3.08, Radix Changii decocting liquid group is 55.573 ± 3.423 (P＜0.01), and Radix Changii polysaccharide group is 51.039 ± 2.724 (P＜0.001).Illustrate that Radix Changii has activating macrophage C
1The b receptor promotes body's immunity.(3) oral administration gavage Radix Changii decocting liquid (5g/Kg, 10g/Kg) and polysaccharide (50mg/Kg, 100mg/Kg).Can increase the thymus index (P＜0.01) and the board index (P＜0.05 or P＜0.01) of normal mouse; Increase mice peripheral white blood cell, lymphocyte number and the lymphocytic percentage rate of ANAE; Promotion is with the phagocytic function (P＜0.05) of shape endothelial system.Antagonism immunosuppressant cyclophosphamide (HC) to the mice peripheral blood from cell and lymphocytic minimizing effect.Can make it to return to normal level.On the other hand, Radix Changii decocting liquid and polysaccharide demonstrate significant inhibitory effect again to the mice delayed allergy due to the dinitrochlorobenzene (DNCB), illustrate that Radix Changii can alleviate the inflammation due to the IV allergy, The above results prompting Radix Changii has certain " strengthening the body resistance " effect, may have potential using value on diseases such as anti-curing cancers and infection.The Radix Changii polysaccharide is one of active component.
2, the anti-stress ability effect studies show that; oral administration gavage bright party decocting liquid and polysaccharide can not only the significant prolongation normal pressure under scarce chlorine survival time of animals; ask about the time that mice at high temperature survives during the survival of the chemical anoxia animal due to can also significant prolongation potassium cyanide; the prompting Radix Changii is a kind of medicine that adapts to the former state effect; but enhancing body has the certain protection effect to the non-specific resistivity of destructive stimulus to body.
3, lipid-lowering effect is given experimental hyperlipidemia rat feeding Radix Changii water extract 2.5g, 5g, 10g/Kg/d) and ethanol extract (125mg, 250mg, 500mg/Kg/d), positive control made with the fat-reducing medicament clofibrate, around the successive administration.The result shows, each administration group all has hypercholesterolemia effect in various degree: the high fat moulding of serum cholesterol group is 681.12 ± 133.53mg%, the clofibrate group is 405.67 ± 204.13mg%, Radix Changii water extract group is 372.75 ± 117.14～447.33 ± l57.66mg%, and Radix Changii ethanol extract group is 471.0 ± 116.19～502.17 ± 116.15mg%.The hypercholesterolemia degree is better than clofibrate (rate of descent is 40.49%) with Radix Changii water extract (2.5g/Kg/d) for best (rate of descent is 45.32%), may with the water extract in to contain the ornithine of more amount relevant, the prompting Radix Changii has the better prevention effect to hypercholesterolemia and atherosclerosis.This experiment is to a little less than reducing serum triglycerides effect, but can improve the ratio of HDL-C/T-CHOL.Infer that thus the Radix Changii effect for reducing blood fat mainly is by suppressing HMGCoA reductase synthetic cholesterol, make the cholesterolemia lowering of concentration, playing effect for reducing blood fat.
4, suppressing blood pulls for a short time and assembles and the blood coagulation resisting function in vitro tests shows, the different extracts energy of Radix Changii significant prolongation man exempts from the blood plasma blood coagulation and strides former time (PT) and thrombin time (TT), the methanolic extract effect is the most obvious, P＜0.01, and can significantly suppress the inductive blood of ADP and pull gathering for a short time, methanolic extract, the platelet aggregation inhibition rate of water extract is respectively 75.30% and 79.75%, with Radix Salviae Miltiorrhizae Injection suitable (77.79%).In vivo test shows that Radix Changii methanolic extract and decocting liquid is energy significant prolongation clotting time of mice all, and the prompting Radix Changii is in the prospect that has further further investigation aspect treatment of vascular intravascular coagulation and the thrombosis.
5, anti-radical action studies show that, the different extracts of Radix Changii generate external rat liver homogenate supernatant lipid peroxide has remarkable inhibitory action, press down with methanolic extract especially the strongest, with matched group relatively, P＜0.001 illustrates that Radix Changii has the effect of anti peroxidation of lipid thing.Methanolic extract and water extract all can improve the activity of superoxide dismutase (SOD), whole blood glutathione peroxidase (GSH-Px) in the hypercholesterolemia rat body, and the blood lipid reducing peroxide produces MDA content.Think that at present free radical can damage cell membrane, cause the generation of multiple senile disease and cause body aging.Radix Changii energy enhancement of SOD and GSH-Px vigor promote the elimination of free radical, suppress lipid peroxide, to certain meaning that has of prevention Senile disease.
6, C antimutagenic effect oral administration gavage Radix Changii water extract 40g/Kg) and methanolic extract (100mg/Kg, 200mg/Kg300mg/Kg),
mMice 10 days, the result shows, group younger sister chromatid interchange (Sister Chromatid Exchange, sCE) frequency: matched group is 2.335, and the cyclophosphamide group is 12.313, and Radix Changii water extract is 7.025, and the Radix Changii methanolic extract is 6.653～7.065.(Micronuclei, MN) frequency: matched group is 2.2 to micronucleus, and the cyclophosphamide group is 8, and Radix Changii water extract is 2.6, and the Radix Changii methanolic extract is 2.8～5.2.Because SCE and MN detect antimutagen sensitivity, reliable index, thereby can think that Radix Changii is inhibited to the SCE frequency that cyclophosphamide brings out rising, cyclophosphamide is brought out the MN frequency of rising, remove the methanol extraction beyond the region of objective existence of 300mg/Kg, all inhibited, can correction of deviation on gene level.
7, other effect research shows that also Radix Changii has remarkable promotion intestinal propulsion effect (P＜0.001), and its action intensity and Rhizoma Dioscoreae are suitable, points out Radix Changii to improving gastrointestinal function, stopping nausea and vomiting by lowering the adverse flow of QI, and aspect such as aid digestion has certain effect.
6, toxicity and mutagenicity test are given mice oral administration gavage Radix Changii decocting liquid 76.25g/Kg, Radix Changii water extract 600gKg, the bright polysaccharide 4.88g/Kg of party observe continuously do not see in 3 days overt toxicity should, anatomic observation does not see that its internal organs have unusually, infer the LD of Radix Changii decocting liquid thus yet after 7 days
50＞76.25g/Kg, the LD of Radix Changii water extract
50＞600g/Kg Radix Changii polysaccharide LD
50＞4.88g/Kg shows that its toxicity is very little.
Mice SCE (SCE) frequency of oral administration gavage Radix Changii decocting liquid (5g/Kg, 10g/Kg, 20g/Kg, 30g/Kg) is respectively 2.691,2.668,2.658,2.676, compare there was no significant difference (P＞0.05) with matched group (2.509): emblem nuclear (MN) frequency also shows similar result.Because SCE and MN detect mutagenic agent sensitivity, reliable index, so think that Radix Changii does not have mutagenic action under this experiment condition, do not have dose-effect relationship yet.
Claims (3)
1, a kind of extracting method of active ingredients from changium root in turn includes the following steps:
(1) at first the Radix Changii root is put and boiled to the no white heart in the boiling water, take out and to scrape off crust, be cut into decoction pieces after drying; It is characterized in that:
(2) get by technical process (1) gained Radix Changii decoction pieces, add 10 times of amount 50%-95% ethanol, placed the multi-function extractor hot reflux 1-2 hour, filtered while hot, add 10 times of amount 50%-95% alcohol heat reflux 0.5-1.5 hour again, filtered while hot, and merging filtrate; Or get by technical process (1) gained Radix Changii decoction pieces, add the 70%-95% ethanol percolation colourless to solution, percolate;
(3) with above-mentioned filtrate or percolate decompression recycling ethanol, gained extractum after drying the Radix Changii ethanol extract.
2, a kind of generation, adjusting immunologic function, mutation, raising stress ability of pressing the active ingredients from changium root of the described method extraction of claim 1 in the lipid peroxide of preparation antithrombotic, blood fat reducing, inhibition hypercholesterolemia improves the application in the intestinal function medicine.
3, a kind ofly contain the medicine that right requires the active ingredients from changium root that 1 described method extracts.
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| CN93111769A CN1047088C (en) | 1993-10-18 | 1993-10-18 | Process for extracting active ingredients from changium root and medicinal use |
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| CN1047088C true CN1047088C (en) | 1999-12-08 |
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| CN1069547C (en) * | 1996-06-18 | 2001-08-15 | 黄德丰 | Gradient extraction technology for extracting multiple active ingredients of medicinal plant |
| CN102525950B (en) * | 2012-02-10 | 2013-06-19 | 兰州大学 | Method for preparing pellets of Se-polysaccharides in Codonopsis pilosula |
| CN103040902B (en) * | 2012-12-13 | 2016-07-20 | 大兴安岭林格贝寒带生物科技股份有限公司 | A kind of method of the wild Radix Codonopsis saponin of purification |
| CN103829123A (en) * | 2014-01-09 | 2014-06-04 | 山西春阳生物科技有限公司 | Method for producing hypoglycemic hypolipemic product by using tartary buckwheat |
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