CN1515565A - Sarcandra flavones coumarins total extract, its medicine composition and application - Google Patents
Sarcandra flavones coumarins total extract, its medicine composition and application Download PDFInfo
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Abstract
The present invention relates to flavone coumarin total extract in the Chinese medicinal material sarcandra twig and leaf, its preparation method and medicine composition containing said total extract. The total extract is obtained by making the Chinese medicinal material sarcandra twig and leaf undergo the processes of alcohol reflux extraction, alcohol diacolation extraction and water decoction extraction. The total extract and its medicine composition can be used for curing primary and secondary thrombocytopenic purpura, auxiliary therapy of carcinomata, raising white blood cell, resisting bacteria and curing the diseases of pneumonia, appendicitis and cellulitis, etc. The composition can be made into the various dosage forms of tablet, granules, capsule, pill, oral liquid, injection and others.
Description
The present invention relates to the field of Chinese medicines, particularly, the present invention relates to Chinese medicinal materials Herba Sarcandrae flavone class coumarins total extract and pharmaceutical composition and purposes.
Sarcandra glaber is Chloranthaceae plant Sarcandra elabra (Thunb.) Nabal, among the peoplely be called Herba Pileae Scriptae, the nine joint bitter edible plant, Williams Elder Twig, refute bone tea, bone wind and disappear etc., be perennial evergreen draft or undershrub, be distributed in provinces such as little, Zhejiang of Chinese river tenth of the twelve Earthly Branches, peace, Hubei, Hunan, Sichuan, Guangdong, Guangxi, it is conventional Chinese medicine simply, be applied to the historical two thousand years of curing the disease clinically, critical role arranged in the Chinese materia medica, herb can expelling wind and clearing away heat, invigorate blood circulation, pain relieving, stimulate the menstrual flow, synthetism.Be used for the treatment of common diseases associated with inflammation, also be used for the treatment of teniasis, rheumatoid arthritis and preventing cold etc.Also be used for the treatment of multiple malignant tumour such as Vipoma, cancer of the stomach, the rectum cancer, liver cancer and esophagus cancer etc.Sarcandra glaber contain fumaric acid, succsinic acid, lignum-vitae base lignan (guaiacyl lignin), Coumarins composition isofraxidin (isofraxidin7-hydroxy-6,8-dimethoxycoumarin), phenols, tannin, flavonoid glycosides (astilbin), coumarin, volatilization wet goods.
China Jiangxi Province is the main product ground of Chinese medicinal material of sarcandra glaber, and resource is very abundant, and the whole province various places have wild and the cultivation product, and in Ganzhou, cities and counties such as Ji'an, Pingxiang have set up the production base, the plantation area reach 50 surplus ten thousand mu, produce per year about 5,000,000 kilograms, formed the staple commodities medicinal material.In view of this, we have carried out long-term further investigation to the Chinese medicine sarcandra glaber, develop efficient, safe, quality controllable sarcandra glaber effective parts formulation, obtained Herba Sarcandrae flavone class, coumarins total extract and pharmaceutical composition thereof, filled up the blank in the sarcandra glaber research.Because the tcm clinical practice medication is with oral the most commonly used, be traditional Chinese medicine injection secondly, therefore, we consider that this Herba Sarcandrae flavone class, coumarins total extract pharmaceutical composition make oral, injection liquid formulation, and finish the present invention thus.
Therefore, the purpose of this invention is to provide Chinese medicine Herba Sarcandrae flavone class coumarins total extract;
Another object of the present invention has provided the method for preparing Herba Sarcandrae flavone class coumarins total extract;
Another object of the present invention has provided the pharmaceutical composition that contains Herba Sarcandrae flavone class coumarins total extract;
Another object of the present invention has provided the purposes that Herba Sarcandrae flavone class coumarins total extract is used to prepare the medicine for the treatment of bleeding due to blood-heat, ecchymosis, primary and secondary thrombocytopenic purpura disease;
Another object of the present invention has provided the purposes that Herba Sarcandrae flavone class coumarins total extract is used to prepare anti-pneumonia, ecphyaditis, cellulitis medicine;
Another object of the present invention has provided the purposes that Herba Sarcandrae flavone class coumarins total extract is used to prepare antibacterials;
Another object of the present invention has provided the purposes that Herba Sarcandrae flavone class coumarins total extract is used to prepare leukocyte increasing, cancer adjuvant therapy medicament.
Pharmaceutical composition of the present invention can use the ordinary method preparation in the pharmacy field, and it can be forms such as tablet, granule, capsule, pill, pill, effervescent tablet, ointment, syrup, injection, oral liquid, mixture, sustained release dosage, controlled release preparation or targeting preparation.
Herba Sarcandrae flavone class coumarins total extract of the present invention can provide following method to prepare.
Technical study and assay
Chinese medicine Herba Sarcandrae flavone class coumarins total extract of the present invention can alcohol reflux extracts by sarcandra glaber is carried out, alcohol percolation extracts or water boiling and extraction obtains, wherein:
The alcohol reflux extracting method is: get the Chinese medicinal material of sarcandra glaber meal, with 40%-95% ethanol 3-10 doubly
Amount is made solvent, floods after 1 hour, and heating and refluxing extraction 2-4 time, each 1-4 hour,
Reclaim ethanol, concentrated solution is through adsorption treatment, and drying promptly gets total extract;
The alcohol percolation extracting method is: get the sarcandra glaber meal, make solvent with 30-90% ethanol, behind the dipping, carry out diacolation with 1-6ml/ minute speed, collect percolate, reclaim ethanol; Concentrated solution is through adsorption treatment, and drying promptly gets total extract;
The decoction extracting method is: get the Chinese medicinal material of sarcandra glaber boiling, and each 1-3 hour, filter, filtrate merges, and concentrates, and concentrated solution is drying to obtain total extract through adsorption treatment.
Preparation method's of the present invention operation steps and characteristics thereof are described below:
One. extract
1. refluxing extraction: get the sarcandra glaber meal---add the 40-95% ethanol that 3-10 doubly measures, flood after 1 hour, heating and refluxing extraction 2-4 time each 1-4 hour, reclaims ethanol and is concentrated into every ml soln and contains the crude drug amount and must not be less than 0.5 gram.
2. diacolation extracts: get the sarcandra glaber meal, the percolation under photograph fluid extract and the extractum item (" 2000 editions appendix IO of Chinese pharmacopoeia), make solvent with 30-90% ethanol, behind the dipping, carry out diacolation with 1-6ml/ minute speed, collect percolate, reclaim ethanol; Be concentrated into every ml soln contain the crude drug amount must not be less than 0.5 the gram.
3. decoct and extract: get the Chinese medicinal material of sarcandra glaber boiling, each 1-3 hour, filter, filtrate merges, and concentrates, and it is about more than 1.01 (55-60 ℃) to be concentrated into relative density.
Assay:
Reference substance solution preparation: the accurate control substance of Rutin 100mg of 120 ℃ of drying under reduced pressure that claim to constant weight, put in the 100ml volumetric flask, add 60% ethanol 70ml, put low-grade fever dissolving in the water-bath, put cold, add 60% alcohol dilution to scale, shake up, the accurate 10ml that draws, put in the 50ml volumetric flask, add water to scale, shake up, promptly.
The typical curve preparation:
The accurate reference substance solution 1.0,2.0,3.0,4.0 of drawing, 5.0, put respectively in the 25ml measuring bottle with 6.0ml, respectively add water 5ml, add 5% sodium nitrite solution 1ml, shake up, placed 6 minutes, add 10% aluminum nitrate solution 1ml, shake up, placed 6 minutes, hydro-oxidation sodium test solution 10ml adds water to scale again, shakes up, placed 15 minutes, and made blank with method, measure optical density at the wavelength place of 500nm according to spectrophotometry with 6ml water, with the optical density is ordinate zou, and concentration is X-coordinate, the drawing standard curve.
Extract the need testing solution preparation of concentrated solution
The need testing solution preparation:
Precision takes by weighing the about 20mg of total extract, puts in the 25ml volumetric flask, adds 70% ethanol 10ml dissolving, and thin up is settled to scale, shakes up, promptly.The accurate 1ml that draws, put in the 25ml measuring bottle, method under the sighting target directrix curve preparation, operate from " respectively adding water 5ml " in accordance with the law, make blank with need testing solution 1ml thin up to 25ml, filtration, get filtrate and measure optical density in accordance with the law, read the weight of rutin the need testing solution from typical curve, calculate, promptly.
After measured, the every gram crude drug of its over-ground part refluxing extraction contains total flavonoid and contains 18.04 milligrams with rutin calculating, diacolation extracts every gram crude drug and contains total flavonoid and calculate with rutin and contain 15.28 milligrams, decocts to extract every gram crude drug and contain total flavonoid and calculate with rutin and contain 12.12 milligrams.
Two. separation and purification
The inventor is surprised to find, and the extract that obtains among the preparation method of the present invention can be removed impurity effectively through separation and purification, improves the activity and the drug effect of total extract significantly, and this has also constituted the obvious progressive technical characterictic of the present invention.The method of separation and purification of the present invention comprises acid heavy method, polymeric amide absorption method and macroreticular resin absorbing method.Below separation and refining method and characteristics thereof are further specified.
1. sour heavy method
Get an amount of volume 25ml from extract concentrated solution, adding an amount of ethanol to concentration is 75%, adds hydrochloric acid then and carries out the heavy experiment of acid, and transferring pH is 2, finds that precipitation is few.
From extract concentrated solution, get an amount of volume 50ml, directly add hydrochloric acid and carry out the heavy experiment of acid, from dropping mode (slowly drip or topple over fast), aspect such as alr mode and speed considers, carrying out different experiments respectively, more finally to transfer pH be 2, till no longer producing precipitation, find all to have precipitation, but precipitation is fine and smooth mud shape, and difficulty is separated with supernatant liquor, the difficult total extract that obtains.
Get an amount of volume 50ml in addition from extract concentrated solution, add the vitriol oil and carry out the heavy experiment of acid, find not have precipitation and occur, solution becomes suspension.
In addition former extraction concentrated solution is diluted to different concentration (every ml soln contains 0.5 gram crude drug and 0.4 gram crude drug) or is concentrated into every ml soln and contain 1 gram crude drug, get an amount of volume 25ml, add hydrochloric acid and carry out the heavy experiment of acid, find all to have precipitation, but precipitation is fine and smooth mud shape, difficulty is separated with supernatant liquor, the difficult total extract that obtains.Scrape and get partly precipitated in furnace pot, oven dry becomes black solid, and surveying it, to contain total flavonoid be 25.18%, and yield is very low.
Therefore, the heavy method separation and purification effect of acid is poor.
2. polymeric amide absorption method
Take by weighing polymeric amide (30-60 order) 6 grams of having handled well, wet method dress post (12 * 25mm), get and extract upward sample of concentrated solution 30ml (containing crude drug 24 grams), the water 400ml of elder generation removes impurity, 30% ethanol 500ml wash-out then, 70% ethanol 400ml wash-out then, respectively 3 kinds of solution concentration are settled to 100ml, as need testing solution, it is carried out the polyamide layer point sample, colour developing, finding has a large amount of flavones ingredients in the water lotion, contain a small amount of flavones ingredient in 30% ethanol eluate, 70% ethanol eluate does not launch in the developping agent of 36% acetic acid, concentrates on initial point.
The accurate 1ml concentrated solution of drawing, put in the 25ml volumetric flask, method under the sighting target directrix curve preparation, operate from " respectively adding water 5ml " in accordance with the law, do blankly with need testing solution 1ml thin up to 25ml, measure optical density in accordance with the law, read the weight of rutin the need testing solution from typical curve, calculate, promptly.Record that to contain total flavonoid in the water lotion be that to contain total flavonoid in 2.27%, 30% ethanol eluate be that to contain total flavonoid in 0.197%, 70% ethanol eluate be 0.048%.
Improve one's methods: reduce applied sample amount, make a living 4 times of dose of make a living 1.5 times of dose of water lotion volume, 30% ethanol eluate volume.Flavonoid mainly concentrates in 30% ethanol eluate as a result.
Get the chromatography column of 2 25mm diameters in addition, respectively take by weighing polymeric amide (30-60 order) 30 grams to handle well, wet method dress post, impurity is removed in each 100ml of applied sample amount (always containing crude drug 160 grams) washing, 30% ethanol eluate recovery ethanol is concentrated into thick paste vacuum-drying and gets total extract, its yield 3.09%, total extract contains total flavonoid 59.73%
3. macroporous adsorbent resin method
Get the chromatography column of 2 25mm diameters, take by weighing each 200 gram of D101 macroporous adsorbent resin of having handled well, wet method dress post, impurity is removed in each 100ml of applied sample amount (always containing crude drug 160 grams) washing earlier, 30% ethanol eluate recovery ethanol is concentrated into thick paste vacuum-drying and gets total extract, its yield 6.17%, total extract contains total flavonoid 55.87%.
(1) comparison of various macroporous adsorbent resins
With the macroporous adsorbent resin such as the H103 of the various models bought from the market, NKA-II, NKA-9, S-8, D3520, D4006, D4020, AB-8, D101 pre-treatment is good, respectively takes by weighing 40 gram resins, wet method dress post (each 40ml of 12 * 25mm) applied sample amounts (always containing crude drug 32 grams), impurity is removed in washing earlier, then make a living 4 times of dose of 30% ethanol elution to cumulative volume.30% ethanol eluate is reclaimed the concentrated 100ml that is settled to of ethanol, as need testing solution, it is carried out the polyamide layer point sample, developping agent is 36% acetic acid, the colour developing of 10% aluminum chloride ethanol, the resin of finding the AB-8 model is to the enrichment better of contained flavones ingredient.
(2) comparison of the applied sample amount of macroporous adsorbent resin
The water lotion of above-mentioned various resins concentrated be settled to 100ml, as need testing solution, it is carried out the colour developing of polyamide layer point sample, find all to have yellow spotting to occur, possible applied sample amount is excessive.Now its different applied sample amounts are compared.
Take by weighing the good D101 resin of pre-treatment 40 gram (3 parts), and wet method dress post (12 * 25mm) 3, applied sample amount is got 20ml respectively, 30ml, 40ml, impurity is removed in washing earlier, then with make a living 4 times of dose of 30% ethanol elution to cumulative volume.With water lotion, 30% ethanol eluate recovery ethanol is concentrated into every ml soln and contains 0.64 gram crude drug, as need testing solution, it is carried out the polyamide layer point sample, developping agent is 36% acetic acid, the colour developing of 10% aluminum chloride ethanolic soln finds that the applied sample amount of 20ml is proper.
Experimental result sees Table 1:
Table 1
Resin 30% alcohol is washed part A value total flavonoid content %
NKA-9 0.605 1.7108
D4006 0.565 1.6002
D4020 0.660 1.8662
NKA-II 0.218 0.1625
D3520 0.855 2.4125
H103 0.659 1.8635
S-8 0.625 0.8841
AB-8 0.997 1.413
D101 0.625 1.767
Therefore, the macroporous adsorbent resin of selecting the AB-8 model is for well, and the resin absorption crude drug amount 16 that per 40 grams have been handled well restrains, make a living 4 times of dose of make a living 15 times of dose of water lotion volume, 30% ethanol eluate, its elutriant reclaims ethanol, evaporate to dryness gets total extract, surveys its total flavonoid content.
Three. the research of drying means and drying temperature
Herba Sarcandrae flavone class after refining, the drying means and the drying temperature of coumarins total extract medicine are studied.Experimental design is as follows: adopt oven drying method, boulton process, three kinds of drying meanss of spray-drying process.Oven drying method, boulton process carry out 5 kinds of differing tempss (50 ℃, 65 ℃, 75 ℃, 85 ℃, 95 ℃) exsiccant respectively relatively.Investigate Herba Sarcandrae flavone class, coumarins content (in dry product).The results are shown in following table.
The measurement result of oven drying method differing temps sees Table 2
Table 2
Total Herba Sarcandrae flavone class, tonka bean camphor
Bake out temperature (℃) drying time (hour)
Class is carried thing medicine content (%)
50 60 52.26
65 52 53.00
75 42 52.23
85 38 56.03
95 36 57.89
The measurement result of boulton process differing temps sees Table 3
Vacuum-drying
Total Herba Sarcandrae flavone class, tonka-bean
Time of drying (hour)
Temperature (℃)
Plain class content (%)
50 46 53.28
65 38 52.89
75 30 52.10
85 26 51.59
95 26 55.28
The measurement result of spray-drying process differing temps sees Table 4
Table 4
Total Herba Sarcandrae flavone class, perfume (or spice) when medicinal material is thrown injection port air outlet drying
(h) legumin class content between sample number material kg temperature temperature ℃
(℃)
1 40 215 133 6 55.0?5％
2 32 175 125 6.5 54.94％
3 25 220 130 5 54.44％
4 30 160 115 7 52.03％
From top table as can be known, oven drying method, vacuum-drying required time are long, must in water-bath, carry out sufficient drying before the vacuum-drying, moisture controlled at 15-20%, easy-to-drawly during vacuum-drying is driedly become cellular, drying can be finished in 2-3 days, otherwise the water content height can overflow during vacuum-drying, and the oven drying method required time is longer, can't adapt to suitability for industrialized production.And the spraying drying time is short, and it is about 1.01 that extracting solution only need be concentrated into relative density, and drying can be finished in a few hours.Because bake drying, vacuum-drying and spraying drying institute time-consuming differ too big, bake drying, vacuum-drying time are spray-dired more than 20 times, therefore, select spraying drying.
Four. stability test
This product is under the circulation of commodities terms of packing, undertaken by continuous three months three lot number samples that room temperature keeps sample, the fixed temperature and humidity accelerated tests detects to different batches, and with 0 month relatively, its appearance character, discriminating, moisture, assay all meet this product quality standard as a result, health examination is also up to specification, illustrates that Herba Sarcandrae flavone class, coumarins total extract bulk drug are basicly stable at the preliminarily stabilised duration of test.
Five. pharmacodynamic experiment
Give rat oral gavage Herba Sarcandrae flavone class, coumarins total extract, can obviously shorten mouse docking bleeding time and clotting time, strengthen the thrombocyte contractile function, antagonism rabbit thrombosis, normal mice, dog platelet counts there is not obvious influence, to the obvious suppression effect being arranged with mouse due to the acid amides of ring phosphorus west, dog thrombocyte, leukopenia and to mouse, dog thrombocyte, leukopenia due to the radiation, mouse, dog thrombocyte, oligoleukocythemia due to benzene, the dimethylbenzene also there are the obvious suppression effect, and make it to recover normal.
Can increase mouse immune organ weight index, the anti-inflammatory action test shows that cape jasmine total extract height, middle dosage group can suppress the ear swelling that dimethylbenzene causes mouse; Suppress the pedal swelling that carrageenin causes mouse; Granuloma induced by implantation of cotton pellets only is inhibition trend, compares no difference of science of statistics with the normal control group.The in-vitro antibacterial test, adopt staphylococcus aureus, 11 kind of 117 strain bacterium such as staphylococcus epidermidis, first suis, coryneform bacteria, diphtheroid, Pseudomonas aeruginosa, pneumobacillus, Bacillus proteus, intestinal bacteria, Candida albicans and bacteroides fragilis shows that through MIC and MBC test the cape jasmine total extract has anti-microbial effect in various degree.
Claims (10)
1. Chinese medicine Herba Sarcandrae flavone class coumarins total extract, it prepares by among the following preparation method one or more:
A. alcohol reflux extracting method: this method is with the Chinese medicinal material of sarcandra glaber meal, doubly measures with 40%-95% ethanol 3-10 and makes solvent, floods after 1 hour, heating and refluxing extraction 2-4 time each 1-4 hour, reclaims ethanol, concentrated solution is handled through separation and purification, and drying promptly gets total extract; Or
B. alcohol percolation extracting method: this method is with the sarcandra glaber meal, makes solvent with 30-90% ethanol, behind the dipping, carries out diacolation with 1-6ml/ minute speed, collects percolate, reclaims ethanol, and concentrated solution is handled through separation and purification, and drying promptly gets total extract; Or
C. decoct extracting method: this method is with the Chinese medicinal material of sarcandra glaber boiling, each 1-3 hour, filter, and filtrate merges, and concentrates, and concentrated solution is handled through separation and purification, and drying promptly gets total extract;
Wherein, the separation and refining method among step a, b or the c is heavy method, polymeric amide absorption method or a macroreticular resin absorbing method of acid.
2. according to the total extract of claim 1, wherein the separation and refining method among step a, b or the c is a macroreticular resin absorbing method.
3. according to the total extract of claim 2, wherein the macroporous resin that uses in the macroreticular resin absorbing method among step a, b or the c is an AB-8 type resin.
4. the preparation method of the Chinese medicine Herba Sarcandrae flavone class coumarins total extract of one of claim 1-3, this method comprises one or more among the following preparation method:
A. alcohol reflux extracting method: this method is with the Chinese medicinal material of sarcandra glaber meal, doubly measures with 40%-95% ethanol 3-10 and makes solvent, floods after 1 hour, heating and refluxing extraction 2-4 time each 1-4 hour, reclaims ethanol, concentrated solution is handled through separation and purification, and drying promptly gets total extract; Or
B. alcohol percolation extracting method: this method is with the sarcandra glaber meal, makes solvent with 30-90% ethanol, behind the dipping, carries out diacolation with 1-6ml/ minute speed, collects percolate, reclaims ethanol, and concentrated solution is handled through separation and purification, and drying promptly gets total extract; Or
C. decoct extracting method: this method is with the Chinese medicinal material of sarcandra glaber boiling, each 1-3 hour, filter, and filtrate merges, and concentrates, and concentrated solution is handled through separation and purification, and drying promptly gets total extract;
Wherein, the separation and refining method among step a, b or the c is heavy method, polymeric amide absorption method or a macroreticular resin absorbing method of acid.
5. the Herba Sarcandrae flavone class coumarins total extract of one of claim 1-3 is used to prepare the purposes of the medicine for the treatment of primary and secondary thrombocytopenic purpura disease.
6. the Herba Sarcandrae flavone class coumarins total extract of one of claim 1-3 is used to prepare the purposes of anti-pneumonia, ecphyaditis, cellulitis medicine.
7. the Herba Sarcandrae flavone class coumarins total extract of one of claim 1-3 is used to prepare the purposes of antibacterials.
8. the Herba Sarcandrae flavone class coumarins total extract of one of claim 1-3 is used to prepare the purposes of leukocyte increasing, cancer adjuvant therapy medicament.
9. pharmaceutical composition, it contains Herba Sarcandrae flavone class coumarins total extract and pharmaceutically acceptable carrier of one of claim 1-3 as activeconstituents.
10. according to the pharmaceutical composition of claim 9, it can be tablet, granule, capsule, pill, pill, effervescent tablet, ointment, syrup, injection, oral liquid, mixture, sustained release dosage, controlled release preparation or targeting preparation.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100374105C (en) * | 2005-12-13 | 2008-03-12 | 南京虹桥医药技术研究所 | Glabrous sarcandra herb lyophilized powder for injection, its high-dose injection and preparing method |
| CN1879672B (en) * | 2006-05-17 | 2010-06-16 | 连晓媛 | Effective components of glabrous sarcandra herb - total polyphenol, its preparation method and application |
| CN101665479B (en) * | 2009-09-22 | 2012-05-16 | 三明华健生物工程有限公司 | Process for synchronously extracting isofraxidin and flavonoid compounds from sarcandra glabra and application thereof |
-
2003
- 2003-01-08 CN CNA031000533A patent/CN1515565A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100374105C (en) * | 2005-12-13 | 2008-03-12 | 南京虹桥医药技术研究所 | Glabrous sarcandra herb lyophilized powder for injection, its high-dose injection and preparing method |
| CN1879672B (en) * | 2006-05-17 | 2010-06-16 | 连晓媛 | Effective components of glabrous sarcandra herb - total polyphenol, its preparation method and application |
| CN101665479B (en) * | 2009-09-22 | 2012-05-16 | 三明华健生物工程有限公司 | Process for synchronously extracting isofraxidin and flavonoid compounds from sarcandra glabra and application thereof |
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