CN109694393B - A kind of extracting method and its application of stachyose - Google Patents
A kind of extracting method and its application of stachyose Download PDFInfo
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Abstract
The present invention relates to a kind of extracting method of stachyose and its applications.The present invention has obtained the higher stachyose of purity, and disclose the application of stachyose after pretreatment, extraction, refluxing extraction, filtering decoloration, ion exchange and concentration.The experimental results showed that the stachyose purity is high that extracting method of the present invention obtains, the advantages that good application effect.
Description
Technical field
The present invention relates to a kind of stachyose extractive technique fields.Specifically the present invention relates to a kind of extracting methods of stachyose.
The invention further relates to the applications of the stachyose.
Background technique
The cancer of the esophagus is common one of malignant tumour, and disease incidence occupies the 5th of all kinds of malignant tumours, and the death rate occupies the 4th
Position.China is the high-incidence country of the cancer of the esophagus, every year there are about 250,000 new diagnosis of esophageal cancer cases, accounts for the hundred of whole world cancer of the esophagus case
/ five ten, every year because of about 150,000 people of death from esophageal carcinoma, percent the 25 of whole mortality of malignant tumors persons are accounted for, (this data is selected from
" Chinese Journal of Traditional Chinese Medicine " the 8th phase in 2018 page 969 and " malignant tumour Chinese medicine practice guidelines " Lin Hongsheng chief editor, the People's Press
Publish page 283), therefore find effective treatment oesophagus cancer drug and write important realistic meaning.
Currently, Chinese medicine plays unique advantage, and drug resource is abundant to the drug research aspect of the cancer of the esophagus, poison is secondary to be made
With small, and there are many action pathway, have broad application prospects.Cancer of the esophagus is clinically attributed to mental disorder and hands over resistance, essence by Chinese medicine
Thanks to heat knot, extravasated blood is interior to be tied, the diseases such as deficiency of QI and declination of YANG (see " combination of Chinese tradiational and Western medicine clinical practice "), denumerable in clinical manifestation to demonstrate,prove and see, so
Clearing heat and detoxicating, dissipating phlegm and keeping the adverse QI downwards, activating microcirculation and removing stasis medicinal are mostly used in treatment, the medicines such as enriching yin are moisturized in strengthening the spleen and replenishing qi.Many Chinese medicines are in the cancer of the esophagus
Complex treatment in have alleviate symptom, mitigate Radiotherapy chemotherapy side effect, reduce postoperative complications the effects of.For not receiving or
Not having operation, Radiotherapy chemotherapy treats the patient with esophageal carcinoma of condition, can play control tumour using pure Chinese traditional treatment, the steady state of an illness,
Life quality is improved, extends the effect of life cycle, but existing research and drug emphasize particularly on different fields in drug effect, the mechanism of action and way
Diameter is also different, therefore the Chinese medicine of number of ways comprehensive therapeutic effect is only preferable treatment oesophagus cancer drug.
It has been investigated that the stachyose in radix rehmanniae recen has effects that treat the cancer of the esophagus.But since its extraction process is complicated,
Extraction product is not pure, and content is low, limits the application of stachyose.
Application for a patent for invention CN105315314A discloses a kind of industrialized preparing process that stachyose is extracted from glutinous rehmannia,
By taking juice, removal of impurities, decoloration, ion exchange, vacuum concentration, sterilization and spray drying, obtains a kind of stachyose purity and be higher than
70% stachyose powder product.The method may be implemented stachyose extraction industrialization, but obtained stachyose purity compared with
It is low, and extraction process is coarse, impurity is more in obtained stachyose.
Application for a patent for invention CN101003551A discloses the production method and utilization glutinous rehmannia production wood of a kind of stachyose
The method of sugar and Catalpol.The method is extracted using low temperature, and extracting solution connects continuously across filters pressing, micro-filtration, ultrafiltration, nanofiltration at low temperature
Continuous membrane filtration processes and activated-charcoal column, large pore resin absorption column, cation exchange resin column, anion-exchange resin column it is continuous
Column chromatography procedure is isolated and purified, and stachyose and Catalpol product are produced.Raw material availability is greatly improved, the energy has been saved.
But the stachyose purity obtained by the method only reaches 50%, and extraction is not thorough, and wastes raw material.
It finding after study, the stachyose extracted through radix rehmanniae recen and other Chinese medicine materials carry out compatible use, for
The cancer of the esophagus is treated with very big facilitation.
Application for a patent for invention CN103816483A discloses a kind of drug and preparation method thereof for treating advanced esophageal carcinoma.
Including 20-30 parts of Radix Codonopsis, 10-15 parts of Rhizoma Atractylodis Macrocephalae, 10-15 parts of orange peel, 20-30 parts of semen coicis, 20-30 parts of Chinese yam, 5-10 parts of Radix Glycyrrhizae,
10-15 parts of Cortex Magnoliae Officinalis, 10-15 parts of Fructus Aurantii, 10-15 parts of Radix Curcumae, 5-10 parts of bulbus fritillariae cirrhosae, 10-15 parts of Longstamen Onion Bulb, 10-15 parts of Snakegourd Fruit, Radix Paeoniae Alba
20-30 parts, 10-15 parts of agalloch eaglewood, 10-15 parts of fingered citron, 10-15 parts of rhizoma arisaematis, 10-15 parts of radix scrophulariae, 10-15 parts of dried orange peel, tuber of pinellia 10-
15 parts, 5-10 parts of fructus amomi, 10-15 parts of inular flower, 10-15 parts of raddle, 10-15 parts of lotus stem, 5-10 parts of the coptis, radix aucklandiae 10-15
Part, 10-15 parts of radix pseudostellariae, 10-15 parts of the root of straight ladybell, 20-30 parts of Radix Rehmanniae, Radix Ophiopogonis 10-20 part, 10-20 parts of Radix Angelicae Sinensis, rhizoma imperatae 10-15
Part, 20-30 parts of oldenlandia diffusa, 20-30 parts of Sculellaria barbata, 1-5 parts of centipede, 5-10 parts of scorpio, 10-20 parts of Radix Salviae Miltiorrhizae, Fructus Forsythiae 10-15
Part, 10-20 parts of paris polyphylla, 10-15 parts of peach kernel, 10-15 parts of safflower, 10-15 parts of olibanum and 10-15 parts of myrrh, can achieve qi-regulating
Reducing phlegm and resolving masses, Yin-nourishing and body fluid promoting dissipating bind, the purpose of activating microcirculation and removing stasis medicinal dissipating bind, it is efficient up to 66%.But its constituent species is various, controls
Therapeutic effect is undesirable and price is higher.
Application for a patent for invention CN102526630A discloses a kind of Chinese medicine of dietotherapy cancer of the esophagus, comprising: the root of Chinese clematis, honeycomb,
Pyrola, Inula britannica chinensis, fructus gleditsiae, thallus laminariae, Snakegourd Fruit, rhizoma acori graminei, dendrobium nobile, Centipeda minima, schizonepeta, the dried immature fruit of citron orange, the root of three-nerved spicebush, madder, bletilla,
Turmeric, Radix Salviae Miltiorrhizae, bush, trigone, oyster, ochre, Radix Astragali, Rhizoma Atractylodis Macrocephalae, wilsonii, Radix Paeoniae Alba and Caulis Spatholobi.Chinese medicine provided by the invention
Overcome the shortcomings of existing simple doctor trained in Western medicine operation, radiotherapy or chemotherapeutic treatment cancer of the esophagus, have Small side effects, good effect, it is at low cost, control
More rate is high, the features such as having no drug resistance, in symptom, sign, clinical efficacy, quality of life and the survival rate side for improving esophageal cancer patients
Face have greater advantage Chinese medicine, by its cooperate operation, radiotherapy, chemotherapeutic treatment, realize raising clinical efficacy, quality of life and
Cure rate.But preparation method is complex, and more carbon alkane alcohol are introduced in preparation process, are easily made to patient body health
At further injuring.
Preferably promote to make currently, there is the treatment cancer of the esophagus in China there are no a kind of method for extracting high purity stachyose
With also without a kind of this composition for treating the cancer of the esophagus, component is simple and effective and preparation method is simple to operation, nontoxic
Composition.Prior art part steps also require further improvement, and formula also needs to be further simplified and improve, steady to reach curative effect
Determine, treat the purpose of cancer of the esophagus.
Summary of the invention
In order to solve the problems in the existing technology, the present invention provides a kind of extracting method of stachyose, including following
Step:
(1) it pre-processes: radix rehmanniae recen clean dry pulverizes and sieves；
(2) it extracts: smashed radix rehmanniae recen is impregnated with ethyl alcohol, successively after Microwave Extraction, heating extraction, sieving,
By the filtrate after sieving by miillpore filter, solid A and extracting solution A are obtained；
(3) refluxing extraction: twice by solid A refluxing extraction；Solid B is obtained through deionized water refluxing extraction for the first time and is mentioned
Liquid B is taken, extracts solid B through alcohol reflux to obtain solid C and extracting solution C for the second time；
(4) filtering decoloration: then combined extract A, B, C mix filtrate with active carbon through ultrafiltration membrance filter, adjust
PH is stood, boiling decoloring, and filtrate D is obtained after filtering；
(5) filtrate D ion exchange: is obtained into filtrate E by macroporous absorbent resin；
(6) it is concentrated: stachyose will be obtained to 0.8-1g crude drug/ml after filtrate E concentration.
Further, in the extracting method, radix rehmanniae recen smashes it through 400-600 mesh in step (1).
Further, in the extracting method, concentration of alcohol is 10%-15% in step (2).
Further, in the extracting method, in step (2), by weight, radix rehmanniae recen: ethyl alcohol=1:15-25.
Further, in the extracting method, the power of Microwave Extraction is 300-500W in step (2).
Further, in the extracting method, microwave extraction time is 40-60s in step (2).
Further, in the extracting method, the heating temperature of heating extraction is 50-70 DEG C in step (2).
Further, in the extracting method, the time of heating extraction is 1-3h in step (2).
Further, in the extracting method, the aperture of miillpore filter is 0.6-0.8 μm in step (2).
Further, in the extracting method, in step (3), first time deionized water refluxing extraction, by weight,
Solid A: water=1:5-8；Reflux temperature is 50-80 DEG C；Reflux ratio is 1-4；Return time is 2-4h.
Further, in the extracting method, in step (3), second of alcohol reflux is extracted, concentration of alcohol 20%-
40%；By weight, solid B: ethyl alcohol=1:1-3；Reflux temperature is 30-60 DEG C；Reflux ratio is 0.5-1；Return time is
2-4h。
Further, in the extracting method, in step (4), the aperture of ultrafiltration membrane is 50kDa-80kDa.
Further, in the extracting method, in step (4), by weight, activated carbon weight: filtrate=0.001-
0.003:1.
Further, in the extracting method, in step (4), pH=4.5-5.5, time of repose 1-3h.
Further, in the extracting method, in step (4), the boiling decoloring time is 1.5-3h.
Further, in the extracting method, in step (5), macroporous absorbent resin type HPD-100, D-101, SD-
One of 300.
Further, in the extracting method, in step (5), by weight, filtrate: resin=1:8-11.
Further, in the extracting method, in step (5), eluant, eluent is 5%-15% ethyl alcohol.
A kind of composition for treating the cancer of the esophagus, the stachyose extracted comprising the above method.
Further, composition includes the stachyose that the above method extracts, and further includes raw sienna, Longstamen Onion Bulb, Snakegourd Fruit, half
Summer, Fructus Aurantii, raw curcuma zedoary, raw trigone, unprocessed leech, peach kernel, gecko, centipede, Radix Salviae Miltiorrhizae, Radix Codonopsis, Prunella vulgaris, Fructus Hordei Germinatus, ramulus cinnamomi, wheat
Winter, asparagus fern, Radix Angelicae Sinensis, raw Radix Astragali, campanulaceae.
Further, composition includes that stachyose is extracted from 9-15 parts of radix rehmanniae recens, further includes 20-30 parts of raw sienna, Longstamen Onion Bulb
15-25 parts, 15-25 parts of Snakegourd Fruit, 6-15 parts of the tuber of pinellia, 6-15 parts of Fructus Aurantii, curcuma zedoary 6-15 parts raw, life trigone 6-15 parts, unprocessed leech 6-
12 parts, 6-15 parts of peach kernel, 6-12 parts of gecko, 6-12 parts of centipede, 6-15 parts of Radix Salviae Miltiorrhizae, 9-20 parts of Radix Codonopsis, 9-15 parts of Prunella vulgaris, raw wheat
6-9 parts of bud, 3-9 parts of ramulus cinnamomi, Radix Ophiopogonis 6-15 part, 6-15 parts of asparagus fern, 9-15 parts of Radix Angelicae Sinensis, Radix Astragali 15-30 parts, 6-12 parts of campanulaceae of life.
A kind of preparation for treating the cancer of the esophagus, including filler, disintegrating agent, lubricant, suspending agent, adhesive, corrigent, sweet tea
Taste agent, matrix further include combinations of the above object.
A kind of preparation method of composition that treating the cancer of the esophagus, comprising the following steps:
(1) it pre-processes: raw sienna being wrapped up with cloth, peach kernel is removed the peel, smashs processing to pieces；
(2) it decocts: by the raw sienna handled well and peach kernel and Longstamen Onion Bulb, Snakegourd Fruit, the tuber of pinellia, Fructus Aurantii, raw curcuma zedoary, raw trigone, life
Leech, gecko, centipede, Radix Salviae Miltiorrhizae, Radix Codonopsis, Prunella vulgaris, Fructus Hordei Germinatus, ramulus cinnamomi, Radix Ophiopogonis, asparagus fern, Radix Angelicae Sinensis, raw Radix Astragali, campanulaceae mixing, add
Water decocts 2 times, filtering；
(3) it is concentrated: merging filtrate twice, stachyose is added, be concentrated to get the composition of the treatment cancer of the esophagus.
Further, the preparation method in step (2), by weight, adds water to cook for the first time, water: medicine=6-10:
1；Decocting time is 25-35min.
Further, the preparation method in step (2), by weight, adds water to cook second, water: medicine=4-8:1；
Decocting time is 15-25min.
Further, the preparation method, in step (2), decocting temperature is 70-90 DEG C.
The composition and preparation, the preparation side of extract, the treatment cancer of the esophagus that the extracting method obtains
Application of the composition that method obtains in preparation treatment oesophagus cancer drug.
Raw sienna compatibility radix rehmanniae recen in prescription of the present invention, Radix Ophiopogonis, asparagus fern, when surrendering the punching of stomach town, nourishing yin and nourishing blood heat-clearing, sharp medicine
Enter stomach；Longstamen Onion Bulb compatibility Snakegourd Fruit, the tuber of pinellia, Fructus Aurantii, campanulaceae activate yang out ruffian, and reducing phlegm and resolving masses is moisturized；Unprocessed leech, peach kernel, raw curcuma zedoary, raw three
Rib, Radix Salviae Miltiorrhizae break degrading the channel, activating microcirculation and removing stasis medicinal；Gecko, centipede, Prunella vulgaris resolving hard lump is clearing heat and detoxicating, except the extravasated blood tumor that disappears is superfluous；Radix Codonopsis,
Radix Astragali, which rises, lifts middle-jiao yang, function of the spleen and stomach, tonifying middle-Jiao and Qi；Ramulus cinnamomi activates yang with helps yang transforming qi；Fructus Hordei Germinatus dispersing liver-energy rises temper.Leech, gecko, Wu in side
Centipede, Prunella vulgaris, curcuma zedoary, trigone, Radix Salviae Miltiorrhizae etc. have anti-tumor function.Our compatibility is exquisite, mutually ties from phlegm silt gas, enriching yin activate yang into
Hand, cuts the interpretation of the cause, onset and process of an illness, and number of ways plays comprehensive therapeutic effect.
The present invention extracts to obtain that stachyose purity is higher, has played very big effect in treatment cancer of the esophagus direction.The present invention
Under the conditions of suitable prescription compatibility, content and suitable preparation method, obtain preferable for esophageal carcinoma therapy effect
Composition.Composition of the present invention has formula simple clear, and preparation method is simple and easy to operate, for esophageal carcinoma therapy effect
It is obviously promoted effect.
Specific embodiment
The invention will now be further described with reference to specific embodiments, the advantages and features of the present invention will be with description and
It is apparent, but these examples are merely exemplary, it is not intended to limit the scope of the present invention in any way.Those skilled in the art
Member it should be understood that without departing from the spirit and scope of the invention can details to technical solution of the present invention and form into
Row modifications or substitutions, but these modifications and replacement are fallen within the protection scope of the present invention.
Embodiment 1
1. the extraction of stachyose
(1) it pre-processes: by 1 part of radix rehmanniae recen clean dry, crushed 400 meshes；
(2) it extracts: 15 part 10% of ethyl alcohol of smashed radix rehmanniae recen is impregnated, after Microwave Extraction, heating extraction,
600 meshes are crossed, the filtering with microporous membrane for being 0.6 μm by aperture by the filtrate after sieving obtains solid A and extracting solution A；Microwave
The power of extraction is 300W, microwave extraction time 60s；Heating extraction temperature is 50 DEG C, extraction time 3h；
(3) refluxing extraction: twice by solid A refluxing extraction；Solid B is obtained through deionized water refluxing extraction for the first time and is mentioned
Liquid B is taken, extracts solid B through 20% alcohol reflux to obtain solid C and extracting solution C for the second time；The condition extracted for the first time are as follows:
By weight, solid A: water=1:5, reflux temperature are 50 DEG C, reflux ratio 1, return time 2h；Second of item extracted
Part are as follows: by weight, solid B: ethyl alcohol=1:3, reflux temperature are 30 DEG C, reflux ratio 0.5, return time 4h；
(4) filtering decoloration: then combined extract A, B, C, the ultrafiltration membrance filter through 50KDa mix filtrate and active carbon
It closes, by weight, activated carbon weight: filtrate=0.001:1 adjusts pH=4.5 and stands 1h, boiling decoloring 1.5h, after filtering
Obtain filtrate D；
(5) ion exchange: filtrate D is adsorbed by D-101 type macroporous absorbent resin, the ethyl alcohol that eluant, eluent is 5%, by weight
Part meter is measured, filtrate: resin=1:8 obtains filtrate E；
(6) it is concentrated: stachyose will be obtained to 0.8g crude drug/ml after filtrate E concentration.
2. the preparation method for treating the composition of the cancer of the esophagus
(1) it pre-processes: 20 parts of raw siennas being wrapped up with cloth, 6 parts of peach kernels are removed the peel, smash processing to pieces；
(2) it decocts: by the raw sienna handled well and peach kernel and 15 parts of Longstamen Onion Bulbs, 15 parts of Snakegourd Fruits, 6 parts of tuber of pinellia, 6 parts of Fructus Aurantiis, 6 parts
Raw curcuma zedoary, 15 portions of raw trigones, 12 parts of unprocessed leech, 12 portions of geckoes, 6 parts of centipedes, 15 parts of Radix Salviae Miltiorrhizaes, 9 parts of Radix Codonopsis, 15 portions of Prunella vulgaris, 6 parts
Fructus Hordei Germinatus, 9 parts of ramulus cinnamomi, 6 parts of Radix Ophiopogonis, 15 parts of asparagus ferns, 9 parts of Radix Angelicae Sinensis, 30 parts of raw Radix Astragalis, 12 parts of campanulaceae mixing, add water to cook 2 times,
Filtering；It decocts for the first time, by weight, water: medicine=6:1, decocting time 25min, decocting temperature is 70 DEG C；Second pan-fried
It boils, by weight, water: medicine=4:1, decocting time 15min, decocting temperature is 90 DEG C；
(3) it is concentrated: merging filtrate twice, the stachyose that 9 parts of radix rehmanniae recens extract is added, is concentrated to get the treatment cancer of the esophagus
Composition.
Embodiment 2
1. the extraction of stachyose
(1) it pre-processes: by 1 part of radix rehmanniae recen clean dry, crushed 600 meshes；
(2) it extracts: 25 part 15% of ethyl alcohol of smashed radix rehmanniae recen is impregnated, after Microwave Extraction, heating extraction,
600 meshes are crossed, the filtering with microporous membrane for being 0.8 μm by aperture by the filtrate after sieving obtains solid A and extracting solution A；Microwave
The power of extraction is 500W, microwave extraction time 40s；Heating extraction temperature is 70 DEG C, extraction time 1h；
(3) refluxing extraction: twice by solid A refluxing extraction；Solid B is obtained through deionized water refluxing extraction for the first time and is mentioned
Liquid B is taken, extracts solid B through 40% alcohol reflux to obtain solid C and extracting solution C for the second time；The condition extracted for the first time are as follows:
By weight, solid A: water=1:8, reflux temperature are 80 DEG C, reflux ratio 4, return time 4h；Second of item extracted
Part are as follows: by weight, solid B: ethyl alcohol=1:1, reflux temperature are 60 DEG C, reflux ratio 1, return time 2h；
(4) filtering decoloration: then combined extract A, B, C, the ultrafiltration membrance filter through 80KDa mix filtrate and active carbon
It closes, by weight, activated carbon weight: filtrate=0.003:1 adjusts pH=5.5 and stands 3h, boiling decoloring 3h, after filtering
Filtrate D；
(5) ion exchange: filtrate D is adsorbed by SD-300 type macroporous absorbent resin, the ethyl alcohol that eluant, eluent is 15% is pressed
Parts by weight meter, filtrate: resin=1:11 obtains filtrate E；
(6) it is concentrated: stachyose will be obtained to 1g crude drug/ml after filtrate E concentration.
2. the preparation method for treating the composition of the cancer of the esophagus
(1) it pre-processes: 30 parts of raw siennas being wrapped up with cloth, 15 parts of peach kernels are removed the peel, smash processing to pieces；
(2) decoct: by the raw sienna handled well and peach kernel and 25 parts of Longstamen Onion Bulbs, 25 parts of Snakegourd Fruits, 15 parts of tuber of pinellia, 15 parts of Fructus Aurantiis,
15 parts of raw curcuma zedoarys, 6 parts of raw trigones, 6 parts of unprocessed leech, 6 portions of geckoes, 12 parts of centipedes, 6 parts of Radix Salviae Miltiorrhizaes, 20 parts of Radix Codonopsis, 9 parts of Prunella vulgaris, 9
Part Fructus Hordei Germinatus, 3 parts of ramulus cinnamomi, 15 parts of Radix Ophiopogonis, 6 parts of asparagus ferns, 15 parts of Radix Angelicae Sinensis, 15 parts of raw Radix Astragalis, 6 parts of campanulaceaes mixing, add water to cook 2
It is secondary, filtering；It decocts for the first time, by weight, water: medicine=10:1, decocting time 35min, decocting temperature is 90 DEG C；Second
Secondary decoction, by weight, water: medicine=8:1, decocting time 25min, decocting temperature is 80 DEG C；
(3) it is concentrated: merging filtrate twice, the stachyose that 15 parts of radix rehmanniae recens extract is added, is concentrated to get treatment oesophagus
The composition of cancer.
Embodiment 3
1. the extraction of stachyose
(1) it pre-processes: by 1 part of radix rehmanniae recen clean dry, crushed 500 meshes；
(2) it extracts: 20 part 10% of ethyl alcohol of smashed radix rehmanniae recen is impregnated, after Microwave Extraction, heating extraction,
600 meshes are crossed, the filtering with microporous membrane for being 0.6 μm by aperture by the filtrate after sieving obtains solid A and extracting solution A；Microwave
The power of extraction is 400W, microwave extraction time 40s；Heating extraction temperature is 60 DEG C, extraction time 2h；
(3) refluxing extraction: twice by solid A refluxing extraction；Solid B is obtained through deionized water refluxing extraction for the first time and is mentioned
Liquid B is taken, extracts solid B through 20% alcohol reflux to obtain solid C and extracting solution C for the second time；The condition extracted for the first time are as follows:
By weight, solid A: water=1:6, reflux temperature are 60 DEG C, reflux ratio 2.5, return time 2h；Second of extraction
Condition are as follows: by weight, solid B: ethyl alcohol=1:2, reflux temperature are 50 DEG C, reflux ratio 0.8, return time 4h；
(4) filtering decoloration: then combined extract A, B, C, the ultrafiltration membrance filter through 50KDa mix filtrate and active carbon
It closes, by weight, activated carbon weight: filtrate=0.002:1 adjusts pH=4.8 and stands 1.5h, boiling decoloring 1.5h, filtering
Filtrate D is obtained afterwards；
(5) ion exchange: filtrate D being adsorbed by HPD-100 type macroporous absorbent resin, the ethyl alcohol that eluant, eluent is 10%,
By weight, filtrate: resin=1:10 obtains filtrate E；
(6) it is concentrated: stachyose will be obtained to 0.9g crude drug/ml after filtrate E concentration.
2. the preparation method for treating the composition of the cancer of the esophagus
(1) it pre-processes: 20 parts of raw siennas being wrapped up with cloth, 6 parts of peach kernels are removed the peel, smash processing to pieces；
(2) it decocts: by the raw sienna handled well and peach kernel and 20 parts of Longstamen Onion Bulbs, 15 parts of Snakegourd Fruits, 6 parts of tuber of pinellia, 8 parts of Fructus Aurantiis, 6 parts
Raw curcuma zedoary, 10 portions of raw trigones, 10 parts of unprocessed leech, 10 portions of geckoes, 6 parts of centipedes, 15 parts of Radix Salviae Miltiorrhizaes, 9 parts of Radix Codonopsis, 12 portions of Prunella vulgaris, 6 parts
Fructus Hordei Germinatus, 9 parts of ramulus cinnamomi, 8 parts of Radix Ophiopogonis, 8 parts of asparagus ferns, 9 parts of Radix Angelicae Sinensis, 22 parts of raw Radix Astragalis, 12 parts of campanulaceae mixing, add water to cook 2 times, mistake
Filter；It decocts for the first time, by weight, water: medicine=8:1, decocting time 25min, decocting temperature is 75 DEG C；Second pan-fried
It boils, by weight, water: medicine=6:1, decocting time 15min, decocting temperature is 80 DEG C；
(3) it is concentrated: merging filtrate twice, the stachyose that 12 parts of radix rehmanniae recens extract is added, is concentrated to get treatment oesophagus
The composition of cancer.
Embodiment 4
1. the extraction of stachyose
(1) it pre-processes: by 1 part of radix rehmanniae recen clean dry, crushed 500 meshes；
(2) it extracts: 20 part 10% of ethyl alcohol of smashed radix rehmanniae recen is impregnated, after Microwave Extraction, heating extraction,
600 meshes are crossed, the filtering with microporous membrane for being 0.6 μm by aperture by the filtrate after sieving obtains solid A and extracting solution A；Microwave
The power of extraction is 400W, microwave extraction time 40s；Heating extraction temperature is 60 DEG C, extraction time 2h；
(3) refluxing extraction: twice by solid A refluxing extraction；Solid B is obtained through deionized water refluxing extraction for the first time and is mentioned
Liquid B is taken, extracts solid B through 20% alcohol reflux to obtain solid C and extracting solution C for the second time；The condition extracted for the first time are as follows:
By weight, solid A: water=1:6, reflux temperature are 60 DEG C, reflux ratio 2.5, return time 2h；Second of extraction
Condition are as follows: by weight, solid B: ethyl alcohol=1:2, reflux temperature are 50 DEG C, reflux ratio 0.8, return time 4h；
(4) filtering decoloration: then combined extract A, B, C, the ultrafiltration membrance filter through 50KDa mix filtrate and active carbon
It closes, by weight, activated carbon weight: filtrate=0.002:1 adjusts pH=4.8 and stands 1.5h, boiling decoloring 1.5h, filtering
Filtrate D is obtained afterwards；
(5) ion exchange: filtrate D being adsorbed by HPD-100 type macroporous absorbent resin, the ethyl alcohol that eluant, eluent is 10%,
By weight, filtrate: resin=1:10 obtains filtrate E；
(6) it is concentrated: stachyose will be obtained to 0.9g crude drug/ml after filtrate E concentration.
2. the preparation method for treating the composition of the cancer of the esophagus
(1) it pre-processes: 20 parts of raw siennas being wrapped up with cloth, 9 parts of peach kernels are removed the peel, smash processing to pieces；
(2) it decocts: by the raw sienna handled well and peach kernel and 20 parts of Longstamen Onion Bulbs, 25 parts of Snakegourd Fruits, 9 parts of tuber of pinellia, 9 parts of Fructus Aurantiis, 9 parts
Raw curcuma zedoary, 9 portions of raw trigones, 9 parts of unprocessed leech, 9 portions of geckoes, 12 parts of centipedes, 12 parts of Radix Salviae Miltiorrhizaes, 15 parts of Radix Codonopsis, 12 portions of Prunella vulgaris, 6 parts
Fructus Hordei Germinatus, 3 parts of ramulus cinnamomi, 9 parts of Radix Ophiopogonis, 9 parts of asparagus ferns, 12 parts of Radix Angelicae Sinensis, 20 parts of raw Radix Astragalis, 9 parts of campanulaceae mixing, add water to cook 2 times, mistake
Filter；It decocts for the first time, by weight, water: medicine=8:1, decocting time 30min, decocting temperature is 75 DEG C；Second pan-fried
It boils, by weight, water: medicine=6:1, decocting time 20min, decocting temperature is 80 DEG C；
(3) it is concentrated: merging filtrate twice, the stachyose that 12 parts of radix rehmanniae recens extract is added, is concentrated to get treatment oesophagus
The composition of cancer.
Comparative example 1
Compared with Example 1, comparative example 1 does not include refluxing extraction process.
1. the extraction of stachyose
(1) it pre-processes: by 1 part of radix rehmanniae recen clean dry, crushed 400 meshes；
(2) it extracts: 15 part 10% of ethyl alcohol of smashed radix rehmanniae recen is impregnated, after Microwave Extraction, heating extraction,
600 meshes are crossed, the filtering with microporous membrane for being 0.6 μm by aperture by the filtrate after sieving obtains solid A and extracting solution A；Microwave
The power of extraction is 300W, microwave extraction time 60s；Heating extraction temperature is 50 DEG C, extraction time 3h；
(4) filtering decoloration: then the ultrafiltration membrance filter by extracting solution A through 50KDa mixes filtrate with active carbon, by weight
Measure part meter, activated carbon weight: filtrate=0.001:1 adjusts pH=4.5 and stands 1h, boiling decoloring 1.5h, liquor B is obtained after filtering；
(5) ion exchange: liquor B is adsorbed by D-101 type macroporous absorbent resin, the ethyl alcohol that eluant, eluent is 5%, by weight
Part meter is measured, filtrate: resin=1:8 obtains liquor C；
(6) it is concentrated: obtaining stachyose to 0.8g crude drug/ml after liquor C is concentrated.
2. the preparation method for treating the composition of the cancer of the esophagus
(1) it pre-processes: 20 parts of raw siennas being wrapped up with cloth, 6 parts of peach kernels are removed the peel, smash processing to pieces；
(2) it decocts: by the raw sienna handled well and peach kernel and 15 parts of Longstamen Onion Bulbs, 15 parts of Snakegourd Fruits, 6 parts of tuber of pinellia, 6 parts of Fructus Aurantiis, 6 parts
Raw curcuma zedoary, 15 portions of raw trigones, 12 parts of unprocessed leech, 12 portions of geckoes, 6 parts of centipedes, 15 parts of Radix Salviae Miltiorrhizaes, 9 parts of Radix Codonopsis, 15 portions of Prunella vulgaris, 6 parts
Fructus Hordei Germinatus, 9 parts of ramulus cinnamomi, 6 parts of Radix Ophiopogonis, 15 parts of asparagus ferns, 9 parts of Radix Angelicae Sinensis, 30 parts of raw Radix Astragalis, 12 parts of campanulaceae mixing, add water to cook 2 times,
Filtering；It decocts for the first time, by weight, water: medicine=6:1, decocting time 25min, decocting temperature is 70 DEG C；Second pan-fried
It boils, by weight, water: medicine=4:1, decocting time 15min, decocting temperature is 90 DEG C；
(3) it is concentrated: merging filtrate twice, the stachyose that 9 parts of radix rehmanniae recens extract is added, is concentrated to get the treatment cancer of the esophagus
Composition.
Comparative example 2
Compared with Example 1, in comparative example 2, the pH of step (4) is not in range, and activated carbon dosage is excessive.
1. the extraction of stachyose
(1) it pre-processes: by 1 part of radix rehmanniae recen clean dry, crushed 400 meshes；
(2) it extracts: 15 part 10% of ethyl alcohol of smashed radix rehmanniae recen is impregnated, after Microwave Extraction, heating extraction,
600 meshes are crossed, the filtering with microporous membrane for being 0.6 μm by aperture by the filtrate after sieving obtains solid A and extracting solution A；Microwave
The power of extraction is 300W, microwave extraction time 60s；Heating extraction temperature is 50 DEG C, extraction time 3h；
(3) refluxing extraction: twice by solid A refluxing extraction；Solid B is obtained through deionized water refluxing extraction for the first time and is mentioned
Liquid B is taken, extracts solid B through 20% alcohol reflux to obtain solid C and extracting solution C for the second time；The condition extracted for the first time are as follows:
By weight, solid A: water=1:5, reflux temperature are 50 DEG C, reflux ratio 1, return time 2h；Second of item extracted
Part are as follows: by weight, solid B: ethyl alcohol=1:3, reflux temperature are 30 DEG C, reflux ratio 0.5, return time 4h；
(4) filtering decoloration: then combined extract A, B, C, the ultrafiltration membrance filter through 50KDa mix filtrate and active carbon
It closes, by weight, activated carbon weight: filtrate=0.01:1 adjusts pH=6 and stands 1h, and boiling decoloring 1.5h must be filtered after filtering
Liquid D；
(5) ion exchange: filtrate D is adsorbed by D-101 type macroporous absorbent resin, the ethyl alcohol that eluant, eluent is 5%, by weight
Part meter is measured, filtrate: resin=1:8 obtains filtrate E；
(6) it is concentrated: stachyose will be obtained to 0.8g crude drug/ml after filtrate E concentration.
2. the preparation method for treating the composition of the cancer of the esophagus
(1) it pre-processes: 20 parts of raw siennas being wrapped up with cloth, 6 parts of peach kernels are removed the peel, smash processing to pieces；
(2) it decocts: by the raw sienna handled well and peach kernel and 15 parts of Longstamen Onion Bulbs, 15 parts of Snakegourd Fruits, 6 parts of tuber of pinellia, 6 parts of Fructus Aurantiis, 6 parts
Raw curcuma zedoary, 15 portions of raw trigones, 12 parts of unprocessed leech, 12 portions of geckoes, 6 parts of centipedes, 15 parts of Radix Salviae Miltiorrhizaes, 9 parts of Radix Codonopsis, 15 portions of Prunella vulgaris, 6 parts
Fructus Hordei Germinatus, 9 parts of ramulus cinnamomi, 6 parts of Radix Ophiopogonis, 15 parts of asparagus ferns, 9 parts of Radix Angelicae Sinensis, 30 parts of raw Radix Astragalis, 12 parts of campanulaceae mixing, add water to cook 2 times,
Filtering；It decocts for the first time, by weight, water: medicine=6:1, decocting time 25min, decocting temperature is 70 DEG C；Second pan-fried
It boils, by weight, water: medicine=4:1, decocting time 15min, decocting temperature is 90 DEG C；
(3) it is concentrated: merging filtrate twice, the stachyose that 9 parts of radix rehmanniae recens extract is added, is concentrated to get the treatment cancer of the esophagus
Composition.
Comparative example 3
Compared with Example 1, comparative example 3 is in step (5), and experiment parameter is not in range.
1. the extraction of stachyose
(1) it pre-processes: by 1 part of radix rehmanniae recen clean dry, crushed 400 meshes；
(2) it extracts: 15 part 10% of ethyl alcohol of smashed radix rehmanniae recen is impregnated, after Microwave Extraction, heating extraction,
600 meshes are crossed, the filtering with microporous membrane for being 0.6 μm by aperture by the filtrate after sieving obtains solid A and extracting solution A；Microwave
The power of extraction is 300W, microwave extraction time 60s；Heating extraction temperature is 50 DEG C, extraction time 3h；
(3) refluxing extraction: twice by solid A refluxing extraction；Solid B is obtained through deionized water refluxing extraction for the first time and is mentioned
Liquid B is taken, extracts solid B through 20% alcohol reflux to obtain solid C and extracting solution C for the second time；The condition extracted for the first time are as follows:
By weight, solid A: water=1:5, reflux temperature are 50 DEG C, reflux ratio 1, return time 2h；Second of item extracted
Part are as follows: by weight, solid B: ethyl alcohol=1:3, reflux temperature are 30 DEG C, reflux ratio 0.5, return time 4h；
(4) filtering decoloration: then combined extract A, B, C, the ultrafiltration membrance filter through 50KDa mix filtrate and active carbon
It closes, by weight, activated carbon weight: filtrate=0.001:1 adjusts pH=4.5 and stands 1h, boiling decoloring 1.5h, after filtering
Obtain filtrate D；
(5) ion exchange: filtrate D is adsorbed by D-101 type macroporous absorbent resin, the ethyl alcohol that eluant, eluent is 30% is pressed
Parts by weight meter, filtrate: resin=1:4 obtains filtrate E；
(6) it is concentrated: stachyose will be obtained to 0.8g crude drug/ml after filtrate E concentration.
2. the preparation method for treating the composition of the cancer of the esophagus
(1) it pre-processes: 20 parts of raw siennas being wrapped up with cloth, 6 parts of peach kernels are removed the peel, smash processing to pieces；
(2) it decocts: by the raw sienna handled well and peach kernel and 15 parts of Longstamen Onion Bulbs, 15 parts of Snakegourd Fruits, 6 parts of tuber of pinellia, 6 parts of Fructus Aurantiis, 6 parts
Raw curcuma zedoary, 15 portions of raw trigones, 12 parts of unprocessed leech, 12 portions of geckoes, 6 parts of centipedes, 15 parts of Radix Salviae Miltiorrhizaes, 9 parts of Radix Codonopsis, 15 portions of Prunella vulgaris, 6 parts
Fructus Hordei Germinatus, 9 parts of ramulus cinnamomi, 6 parts of Radix Ophiopogonis, 15 parts of asparagus ferns, 9 parts of Radix Angelicae Sinensis, 30 parts of raw Radix Astragalis, 12 parts of campanulaceae mixing, add water to cook 2 times,
Filtering；It decocts for the first time, by weight, water: medicine=6:1, decocting time 25min, decocting temperature is 70 DEG C；Second pan-fried
It boils, by weight, water: medicine=4:1, decocting time 15min, decocting temperature is 90 DEG C；
(3) it is concentrated: merging filtrate twice, the stachyose that 9 parts of radix rehmanniae recens extract is added, is concentrated to get the treatment cancer of the esophagus
Composition.
Comparative example 4
Compared with Example 1, when prepared by 4 composition of comparative example, constituent content goes beyond the scope.
1. the extraction of stachyose
(1) it pre-processes: by 1 part of radix rehmanniae recen clean dry, crushed 400 meshes；
(2) it extracts: 15 part 10% of ethyl alcohol of smashed radix rehmanniae recen is impregnated, after Microwave Extraction, heating extraction,
600 meshes are crossed, the filtering with microporous membrane for being 0.6 μm by aperture by the filtrate after sieving obtains solid A and extracting solution A；Microwave
The power of extraction is 300W, microwave extraction time 60s；Heating extraction temperature is 50 DEG C, extraction time 3h；
(3) refluxing extraction: twice by solid A refluxing extraction；Solid B is obtained through deionized water refluxing extraction for the first time and is mentioned
Liquid B is taken, extracts solid B through 20% alcohol reflux to obtain solid C and extracting solution C for the second time；The condition extracted for the first time are as follows:
By weight, solid A: water=1:5, reflux temperature are 50 DEG C, reflux ratio 1, return time 2h；Second of item extracted
Part are as follows: by weight, solid B: ethyl alcohol=1:3, reflux temperature are 30 DEG C, reflux ratio 0.5, return time 4h；
(4) filtering decoloration: then combined extract A, B, C, the ultrafiltration membrance filter through 50KDa mix filtrate and active carbon
It closes, by weight, activated carbon weight: filtrate=0.001:1 adjusts pH=4.5 and stands 1h, boiling decoloring 1.5h, after filtering
Obtain filtrate D；
(5) ion exchange: filtrate D is adsorbed by D-101 type macroporous absorbent resin, the ethyl alcohol that eluant, eluent is 5%, by weight
Part meter is measured, filtrate: resin=1:8 obtains filtrate E；
(6) it is concentrated: stachyose will be obtained to 0.8g crude drug/ml after filtrate E concentration.
2. the preparation method for treating the composition of the cancer of the esophagus
(1) it pre-processes: 40 parts of raw siennas being wrapped up with cloth, 6 parts of peach kernels are removed the peel, smash processing to pieces；
(2) it decocts: by the raw sienna handled well and peach kernel and 10 parts of Longstamen Onion Bulbs, 30 parts of Snakegourd Fruits, 2 parts of tuber of pinellia, 20 parts of Fructus Aurantiis, 6
Part raw curcuma zedoary, 20 parts of raw trigones, 12 parts of unprocessed leech, 12 portions of geckoes, 6 parts of centipedes, 15 parts of Radix Salviae Miltiorrhizaes, 5 parts of Radix Codonopsis, 15 parts of Prunella vulgaris, 4
Part Fructus Hordei Germinatus, 9 parts of ramulus cinnamomi, 6 parts of Radix Ophiopogonis, 18 parts of asparagus ferns, 9 parts of Radix Angelicae Sinensis, 30 parts of raw Radix Astragalis, 15 parts of campanulaceaes mixing, add water to cook 2
It is secondary, filtering；It decocts for the first time, by weight, water: medicine=6:1, decocting time 25min, decocting temperature is 70 DEG C；Second
Secondary decoction, by weight, water: medicine=4:1, decocting time 15min, decocting temperature is 90 DEG C；
(3) it is concentrated: merging filtrate twice, the stachyose that 18 parts of radix rehmanniae recens extract is added, is concentrated to get treatment oesophagus
The composition of cancer.
Comparative example 5
Compared with Example 1, when preparing composition, experiment parameter goes beyond the scope comparative example 5.
1. the extraction of stachyose
(1) it pre-processes: by 1 part of radix rehmanniae recen clean dry, crushed 400 meshes；
(2) it extracts: 15 part 10% of ethyl alcohol of smashed radix rehmanniae recen is impregnated, after Microwave Extraction, heating extraction,
600 meshes are crossed, the filtering with microporous membrane for being 0.6 μm by aperture by the filtrate after sieving obtains solid A and extracting solution A；Microwave
The power of extraction is 300W, microwave extraction time 60s；Heating extraction temperature is 50 DEG C, extraction time 3h；
(3) refluxing extraction: twice by solid A refluxing extraction；Solid B is obtained through deionized water refluxing extraction for the first time and is mentioned
Liquid B is taken, extracts solid B through 20% alcohol reflux to obtain solid C and extracting solution C for the second time；The condition extracted for the first time are as follows:
By weight, solid A: water=1:5, reflux temperature are 50 DEG C, reflux ratio 1, return time 2h；Second of item extracted
Part are as follows: by weight, solid B: ethyl alcohol=1:3, reflux temperature are 30 DEG C, reflux ratio 0.5, return time 4h；
(4) filtering decoloration: then combined extract A, B, C, the ultrafiltration membrance filter through 50KDa mix filtrate and active carbon
It closes, by weight, activated carbon weight: filtrate=0.001:1 adjusts pH=4.5 and stands 1h, boiling decoloring 1.5h, after filtering
Obtain filtrate D；
(5) ion exchange: filtrate D is adsorbed by D-101 type macroporous absorbent resin, the ethyl alcohol that eluant, eluent is 5%, by weight
Part meter is measured, filtrate: resin=1:8 obtains filtrate E；
(6) it is concentrated: stachyose will be obtained to 0.8g crude drug/ml after filtrate E concentration.
2. the preparation method for treating the composition of the cancer of the esophagus
(1) it pre-processes: 20 parts of raw siennas being wrapped up with cloth, 6 parts of peach kernels are removed the peel, smash processing to pieces；
(2) it decocts: by the raw sienna handled well and peach kernel and 15 parts of Longstamen Onion Bulbs, 15 parts of Snakegourd Fruits, 6 parts of tuber of pinellia, 6 parts of Fructus Aurantiis, 6 parts
Raw curcuma zedoary, 15 portions of raw trigones, 12 parts of unprocessed leech, 12 portions of geckoes, 6 parts of centipedes, 15 parts of Radix Salviae Miltiorrhizaes, 9 parts of Radix Codonopsis, 15 portions of Prunella vulgaris, 6 parts
Fructus Hordei Germinatus, 9 parts of ramulus cinnamomi, 6 parts of Radix Ophiopogonis, 15 parts of asparagus ferns, 9 parts of Radix Angelicae Sinensis, 30 parts of raw Radix Astragalis, 12 parts of campanulaceae mixing, add water to cook 2 times,
Filtering；It decocts for the first time, by weight, water: medicine=1:1, decocting time 50min, decocting temperature is 50 DEG C；Second pan-fried
It boils, by weight, water: medicine=10:1, decocting time 10min, decocting temperature is 90 DEG C；
(3) it is concentrated: merging filtrate twice, the stachyose that 9 parts of radix rehmanniae recens extract is added, is concentrated to get the treatment cancer of the esophagus
Composition.
The measurement of 1 stachyose purity of experimental example
Using the purity of HPLC method measurement stachyose:
Test condition: Waters515 high performance liquid chromatograph is used；Mobile phase: acetonitrile-water (70:30)；Flow velocity: 1ml/
min；Chromatographic column: ShodexAsahipakNH2P-50 (250mm × 4.6mm, 5 μm).
The determination of the range of linearity: drawing 1,2,5,10,12.5,15 μ L of stachyose reference substance solution, injects efficient liquid phase instrument,
Using sample volume as abscissa, integrating peak areas value is that ordinate draws standard curve, obtains regression equation y=8426.5x-
3650.4,r2=0.998, the results showed that, sample volume is good in 12.3-154.8 μ L linear relationship.
After tested, as a result such as table 1:
1 stachyose purity test result of table
| Group | Embodiment 1 | Embodiment 2 | Comparative example 1 | Comparative example 2 | Comparative example 3 |
| Purity (%) | 95% | 94% | 62% | 76% | 70% |
2 composition toxicity test of experimental example
SD rat is selected, the combination of the embodiment of the present invention 1 of various concentration (15.0,10.0,5.0g crude drug/kg) is given
Object is dissolved in distilled water, and SD rat is given in stomach-filling, and daily stomach-filling is primary, and continuous 30 days, 24 hours each groups were living after the last administration
1/2 animal (half male and half female) is killed, work is killed after remaining 1/2 animal continues observation 2 weeks.The appearance, general of animal is observed during test
Behavior, food intake, weight change, 30 days and the 2 weeks progress hematologies of being discontinued after administration (RBC, HB, granulophilocyte, PLT and are divided
Class) and the indexs such as blood biochemical (AST, ALT, ALP, Glu, BUN, Crca, TP), urine biochemistry, organ coefficient, Histopathology
It checks.Test result shows: composition of the invention is good in high, medium and low dosage group animal general state, appearance sign, row
For activity, food-intake and body weight increase variation without exception；Three dosage groups and control group hematological examination, blood biochemical analysis,
Urine biochemical analysis is without significant difference between normal range (NR), group；Each group main organs histopathological examination has no obvious different
Often.These parameters also have no change after being discontinued 2 weeks.The dose of this test is respectively 150,100, the 50 of clinical medicine dose
Times, it is 30 days continuous in high, medium and low three dosage (15.0,10.0,5.0g crude drug/kg) according to test result Chinese medicine of the invention
Administration has no significant effect rat, and without specific toxicity target organ and sensitive indicator, convalescence observation also has no retardance toxicity
Reaction, shows that the dose safety of composition clinical application of the invention is higher.
The measurement of 3 composition curative effect of experimental example
1. Establishment of mouse model
Purchase healthy SD rat 280, half male and half female, weight is in 55-60g, in the lab, 22-25 DEG C of room temperature, constant
Humidity 25-40%, under the conditions of every 12 hours light dark cycles are primary, raising rat after a week, is built using induction property cancer of the esophagus model
Legislation is tested.It is induced using Methyl jasmonate stomach-filling, dosage is 1g/kg weight, 2 times a week, induces 2 weeks, passes through
Check discovery, all rats were induced to form papilloma and cancer in two weeks.280 rats are divided into following 7 groups at random:
Blank group: 40 rats, half male and half female normally feed mouse grain and drinking water；
Example 1 group: 40 rats, half male and half female after feeding mouse grain daily, are pressed by the dosage stomach-filling of 10g/kg weight
The medicine being prepared according to method in embodiment 1；
1 group: 40 rat of comparative example, half male and half female after feeding mouse grain daily, are pressed by the dosage stomach-filling of 10g/kg weight
The medicine being prepared according to method in comparative example 1；
2 groups: 40 rats of comparative example, half male and half female after feeding mouse grain daily, are pressed by the dosage stomach-filling of 10g/kg weight
The medicine being prepared according to method in comparative example 2；
3 groups: 40 rats of comparative example, half male and half female after feeding mouse grain daily, are pressed by the dosage stomach-filling of 10g/kg weight
The medicine being prepared according to method in comparative example 3；
4 groups: 40 rats of comparative example, half male and half female after feeding mouse grain daily, are pressed by the dosage stomach-filling of 10g/kg weight
The medicine being prepared according to method in comparative example 4；
5 groups: 40 rats of comparative example, half male and half female after feeding mouse grain daily, are pressed by the dosage stomach-filling of 10g/kg weight
The medicine being prepared according to method in comparative example 5.
2. Germicidal efficacy
In experimentation, food-intake, the state of mind, the survival condition of rat are observed daily, is recorded weekly, feed 4 weeks.
3. mouse is drawn materials
Taken after 4 weeks, under aseptic condition mouse swallow be cut into taeniae to the oesophagus of stomach after tile, along pharynx to the direction of stomach oesophagus
Tissue is rolled along cross section again, is fixed with needle, is put into and date and organization name are fixed and marked in 10% neutral formalin.
4.HE dyeing observation epithelium of esophagus Histopathology detection
To be put into the esophageal tissue for 24 hours or more being fixed in 10% neutral formalin, conventional organization processing, paraffin embedding, 4 μ
M slice is put into 65 DEG C of ovens and bakes 1h；The paraffin section of drying is immersed into dimethylbenzene, dewax 10min, with washes of absolute alcohol 3
Secondary, deionized water is cleaned 3 times；It is put into Harris hematoxylin dye liquor 5min, is rinsed 3 times with deionized water；Then with 1% hydrochloric acid-
Alcoholic solution breaks up 20s, is flushed three times with deionized water；It is put into 1% ammonium hydroxide and returns blue 3min, deionization is washed 3 times；0.5% alcohol
Dissolubility Yihong liquid dyes 30s, and 80% ethanol solution cleans 3 times；Dimethylbenzene transparent processing 5min, neutral gum mounting；Aobvious
Micro- microscopic observation: the dyeing of nucleus basophilla is blue, and eosinophilic cytoplasmic dyeing takes on a red color.
According to Pathologic Grading, You Liangwei pathologists are under double blind state, optical microphotograph microscopic observation mouse oesophagus
Histopathologic changes are simultaneously divided into: slight atypical hyperplasia (referred to as " light "), moderate atypical hyperplasia (referred to as " in "), severe is not
Typical hyperplasia (referred to as " weight ") and dermoid cancer (referred to as " cancer ").
Normal esophageal squamous cell (0 grade): the orderly proper alignment of Esophagus, squamous epithelial layer thin and thick
Uniformly, core is rounded or oval, and basal cell layer is regular；
Slight atypical hyperplasia (1 grade): slight atypical hyperplasia, special-shaped epithelial cell involve the 1/3 of epithelial layer；
Moderate atypical hyperplasia (2 grades): the 2/3 of epithelium holostrome is involved in special-shaped epithelial cell middle layer；
Severe atypical hyperplasia (3 grades): special-shaped epithelium almost involves holostrome, i.e., 2/3 or more, but still there is thin maturation on surface layer
Squamous cell；
Dermoid cancer (4 grades): cancerous tissue is blocky in piece, and cancerous tissue is in polygonal, core circle or oval, contaminates deeply,
Positioned at cell center, core increases, and endochylema is reduced.
Experimental result such as table 2:
2 curative effect test result of table
Through analyzing, between each group, P < 0.05 has statistical significance.
Through comparing, the composition of the application has good effect in terms for the treatment of the cancer of the esophagus.
The technical means disclosed in the embodiments of the present invention is not limited only to above-mentioned disclosed technological means, further includes by the above skill
Technical solution composed by art feature any combination.The foregoing is a specific embodiment of the present invention, it is noted that for this
For the those of ordinary skill of technical field, without departing from the principle of the present invention, if the improvement can also be made and repaired
Decorations, these also will be regarded as protection scope of the present invention.
Claims (7)
1. a kind of extracting method of stachyose, which comprises the following steps:
(1) it pre-processes: radix rehmanniae recen clean dry pulverizes and sieves；
(2) it extracts: smashed radix rehmanniae recen is impregnated with ethyl alcohol, successively after Microwave Extraction, heating extraction, sieving, incited somebody to action
Filtrate after sieve obtains solid A and extracting solution A by miillpore filter；
(3) refluxing extraction: twice by solid A refluxing extraction；Solid B and extracting solution are obtained through deionized water refluxing extraction for the first time
B extracts solid B through alcohol reflux to obtain solid C and extracting solution C for the second time；
(4) filtering decoloration: then combined extract A, B, C mix filtrate with active carbon through ultrafiltration membrance filter, and it is quiet to adjust pH
It sets, boiling decoloring, filtrate D is obtained after filtering；
(5) filtrate D ion exchange: is obtained into filtrate E by macroporous absorbent resin；
(6) it is concentrated: stachyose will be obtained to 0.8-1g crude drug/ml after filtrate E concentration；
In the step (3), flow back for the first time, by weight, solid A: deionized water=1:5-8, reflux ratio 1-4；
In the step (3), second of reflux, by weight, solid B: ethyl alcohol=1:1-3, reflux ratio 0.5-1；
In the step (4), pH=4.5-5.5, by weight, activated carbon weight: filtrate=0.001-0.003:1.
2. extracting method according to claim 1, which is characterized in that in the step (2), concentration of alcohol 10-15%,
By weight, radix rehmanniae recen: ethyl alcohol=1:15-25, heating temperature are 50-70 DEG C.
3. extracting method according to claim 1, which is characterized in that in the step (2), the power of Microwave Extraction is
300-500W, extraction time 40-60s.
4. extracting method according to claim 1, which is characterized in that in the step (5), macroporous absorbent resin type is
One of HPD-100, D-101, SD-300.
5. extracting method according to claim 1, which is characterized in that in the step (5), by weight, filtrate: tree
Rouge=1:8-11.
6. a kind of composition for treating the cancer of the esophagus, which is characterized in that extract to obtain comprising any method of claim 1-5
Stachyose.
7. composition described in the extract or 6 that any extracting method of claim 1-5 obtains treats oesophagus in preparation
Application in cancer drug.
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