CN102302542A - Chinese medicinal oral liquid for treating cancers - Google Patents
Chinese medicinal oral liquid for treating cancers Download PDFInfo
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- CN102302542A CN102302542A CN201110277502A CN201110277502A CN102302542A CN 102302542 A CN102302542 A CN 102302542A CN 201110277502 A CN201110277502 A CN 201110277502A CN 201110277502 A CN201110277502 A CN 201110277502A CN 102302542 A CN102302542 A CN 102302542A
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- 239000007788 liquid Substances 0.000 title claims abstract description 53
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- 238000000034 method Methods 0.000 claims abstract description 48
- 238000002360 preparation method Methods 0.000 claims abstract description 30
- 239000000284 extract Substances 0.000 claims abstract description 16
- 235000002789 Panax ginseng Nutrition 0.000 claims abstract description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 48
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- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 36
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Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention provides a Chinese medicinal oral liquid for treating cancers. In the oral liquid, extracts of red ginseng, astragalus mongholicus, liquorice, Chinese blister beetle and toad venom serve are used as active components. The invention also provides a preparation method of the Chinese medicinal oral liquid and a method for determining the contents of the active components.
Description
Technical field
The present invention relates to a kind of Chinese medicine oral liquid of treating cancer, said oral liquid comprises the extract of Radix Ginseng Rubra, the Radix Astragali, Radix Glycyrrhizae, Mylabris and Venenum Bufonis as effective ingredient.
Background technology
Existing treatment of cancer has many methods, mostly adopts the bonded mode of Chinese medicine and western medicine to carry out.For example, the method for subclavian vein puncture gives the patient with intravenous injection, and employed medicament mostly is usually used General Medicine, and its curative effect is relatively poor, and tumor regression is slow, and stable disease is poor, even worsens; In addition, adopt the pure chemistry treatment, patient's pain is difficult to be eased, and sleep improves less, and appetite loses increase.
From medically saying; The immunologic function of the generation of malignant tumor, development and human body has very confidential relation; Show through investigation; Immunologic function in patients with malignant tumor, especially cellular immune function are suppressed; Those skilled in the art is devoted to seek the various adjustments (BRM) that can improve biological respinse; To strengthen adjustment or protection immunological function of cancer patients; The adjustment that can improve biological respinse (BRM) that at present has been developed into has a lot, but relatively poor to the therapeutic effect of tumor patient.
In addition, radiotherapy and most chemicals in the existing oncotherapy all can cause suitable bone marrow depression, and this has limited the increase that continues to carry out and treat intensity of effective treatment to a certain extent.
The known tcm product of being processed by Radix Ginseng Rubra, the Radix Astragali, Mylabris and Venenum Bufonis of prior art (DELISHENG ZHUSHEYE) has active anticancer; Result for retrieval in " Chinese drug patent data base " is following:
Summary of the invention
The present invention provides a kind of Chinese medicine oral liquid of treating cancer, and said oral liquid can strengthen adjusting immunological function of cancer patients, and the pain of reduction of patient is improved patient's situation, suppresses the development of tumor, improves the toleration of patient to chemotherapy; This oral liquid comprises the extract of Radix Ginseng Rubra, the Radix Astragali, Radix Glycyrrhizae, Mylabris, Venenum Bufonis as effective ingredient.
The present invention also provides the method for preparing of said Chinese medicine oral liquid, comprises Radix Ginseng Rubra, the Radix Astragali, Radix Glycyrrhizae, Mylabris, Venenum Bufonis are used earlier alcohol reflux, residue reuse water boiling and extraction; This method for preparing can be with basic extraction such as main anticancer component cantharidin, indoles, bufalin, saponin, polysaccharide fully.
The present invention also provides the active constituent content measuring method of said Chinese medicine oral liquid, comprises the content assaying method of total saponins, polysaccharide, cinobufagin and bufogenin, cantharidin, water solublity indoles.
Therefore, the present invention relates to:
(1) a kind of Chinese medicine oral liquid of treating cancer, its effective ingredient is processed by Radix Ginseng Rubra 60-240 weight portion, Radix Astragali 345-1380 weight portion, Radix Glycyrrhizae 96-384 weight portion, Mylabris 1-4 weight portion, Venenum Bufonis 0.5-2 weight portion;
(2) according to the Chinese medicine oral liquid of (1), its effective ingredient is processed by Radix Ginseng Rubra 120 weight portions, the Radix Astragali 690 weight portions, Radix Glycyrrhizae 192 weight portions, Mylabris 2 weight portions, Venenum Bufonis 1 weight portion;
(3), comprise Radix Ginseng Rubra, the Radix Astragali, Radix Glycyrrhizae, Mylabris, Venenum Bufonis are used earlier alcohol reflux, residue reuse water boiling and extraction like the method for preparing of Chinese medicine oral liquid as described in (1);
(4) according to the method for preparing of (3), comprise Radix Ginseng Rubra, the Radix Astragali, Radix Glycyrrhizae are pulverized, add 75% alcohol reflux, obtain extracting solution A; The residue decocte with water is extracted, and adds diluted alkaline to extracting solution and regulates pH＞8, filters, and filtrating is regulated pH 6.5-7.0 with diluted acid, and concentrating under reduced pressure obtains extracting solution B;
Mylabris, Venenum Bufonis are pulverized, added 95% alcohol reflux, obtain extracting solution C; The residue decocte with water is extracted, and filters, and filtrate decompression concentrates, and obtains extracting solution D;
With above extracting solution A, B, C, D mix homogeneously, process oral liquid;
(5) according to the method for preparing of (4), comprise that it is coarse powder that Radix Ginseng Rubra, the Radix Astragali, Radix Glycyrrhizae are pulverized, add 8 times of amount 75% ethanol, reflux, extract, 3 times, each 1.5 hours, merge alcohol extract, filter, filtrate recycling ethanol to nothing alcohol flavor obtains extracting solution A; Add 8 times of water gagings to residue and decoct 3 times, each 20 minutes, merge decocting liquid; Add the calcium hydroxide breast and regulate pH 8.0, add 10% sodium hydroxide solution and regulate pH 9.0, filter; Filtrating is regulated pH 6.5-7.0 with dilute hydrochloric acid, and filtrate decompression concentrates, and obtains extracting solution B;
It is coarse powder that Mylabris is pulverized, and Venenum Bufonis is pulverized and is to mix fine powder, adds 7 times of amount 95% ethanol, reflux, extract, 3 times, and each 20 minutes, the merging alcohol extract, concentrating under reduced pressure obtains extracting solution C; Add 7 times of water gagings to residue and decoct 3 times, each 20 minutes, merge decocting liquid, left standstill 2 hours, to filter, filtrate decompression concentrates, and obtains extracting solution D;
Above extracting solution A, B, C, D are mixed, stir, the embedding of adjustment volume is in infusion bottle, and 100 ℃ of sterilizations 30 minutes were left standstill more than 1 week, filtered, and added the sucrose mixing to filtrating, thin up to 2 times, and embedding is in oral liquid bottle, and sterilization promptly gets;
(6), comprise the content assaying method of total saponins, polysaccharide, cinobufagin and bufogenin, cantharidin, water solublity indoles like the active constituent content measuring method of Chinese medicine oral liquid as described in (1);
(7) according to the content assaying method of (6), comprising:
The content assaying method of a, total saponins:
Measure according to ultraviolet spectrophotometry (2000 editions one appendix V A of Chinese Pharmacopoeia);
The preparation precision of reference substance solution is got red ginseng saponin Re reference substance 8.0mg, and accurate the title decides, and puts in the 5ml volumetric flask, adds methanol and makes dissolving in right amount, and be diluted to scale, shakes up promptly to get, and every milliliter contains red ginseng saponin Re 1.6mg;
The preparation precision of standard curve is measured reference substance solution 20,40,60,80,100ul, puts respectively in the 10ml tool plug test tube, puts and waves most solvent in the water-bath, takes out cooling immediately; The accurate glacial acetic acid solution 0.2ml that adds 5% vanillin, perchloric acid 0.8ml shakes up, and puts heating taking-up in 15 minutes in 60 ℃ of water-baths, immediately with circulating water cooling 2 minutes; The accurate again glacial acetic acid solution 5ml that adds shakes up, and is blank with the corresponding reagent; According to 33 pages of appendix of spectrophotography Pharmacopoeia of the People's Republic of China version in 2000; Wavelength at 544nm is measured trap, is vertical coordinate with the trap, and concentration is the standard curve that abscissa is drawn solution;
Accurate Radix Ginseng Rubra, the Radix Astragali, the Radix Glycyrrhizae semi-finished product concentrated solution 0.5ml of drawing of the preparation of need testing solution puts in the 100ml measuring bottle, and thin up is to scale, close plug, shakes up; The above-mentioned solution 10ml of accurate again absorption; Put in the separatory funnel, with water saturation n-butanol extraction three times, each 20ml; Merge butanol extraction liquid; With washing twice, each 10ml discards water liquid; Evaporate to dryness in the water-bath of n-butyl alcohol liquid; Residue is put in the 5ml measuring bottle with dissolve with methanol, and is diluted to scale, shakes up, promptly gets;
The accurate need testing solution 40ul that draws of algoscopy; Put in the 10ml tool plug test tube; Put and wave most solvent in the water-bath; Take out cooling immediately; The sighting target directrix curve prepares method down from " the vanillin glacial acetic acid solution 0.2ml...... of precision adding 5% ", measures trap in accordance with the law, reads the content of Radix Ginseng Rubra total saponins the need testing solution from standard curve; Calculate, promptly get;
B, content Determination of Polysaccharide method
Measure according to spectrophotography (appendix V of Chinese Pharmacopoeia version in 2000);
The preparation precision of reference substance solution takes by weighing the anhydrous glucose 80mg that is dried to constant weight through 80 ℃, puts in the 100ml volumetric flask, is dissolved in water, and is diluted to scale; Shake up, precision is measured 5ml, puts in the 50ml volumetric flask; Thin up is to scale, shakes up promptly to get, and per 1 milliliter contains anhydrous glucose 0.08mg;
The preparation precision of standard curve is measured reference substance solution 0.2,0.4,0.6,0.8,1.0ml; Put respectively in the 10ml tool plug test tube; Each is managed precision and adds water and make into 1ml; In ice bath, add sulphuric acid anthrone solution 4ml respectively; Shake up; Put in the boiling water bath and heated 10 minutes; Take out; After placing room temperature; With the corresponding reagent is blank, according to appendix of spectrophotography Pharmacopoeia of the People's Republic of China version in 2000, measures trap in the wavelength of 625nm; With the trap is vertical coordinate, and concentration is the standard curve that abscissa is drawn solution; Said sulphuric acid anthrone solution is like this preparation: get anthrone 50mg, add water 25ml and sulphuric acid 75ml makes dissolving, promptly get, face and use stylish preparation;
The water decoction of Radix Ginseng Rubra, the Radix Astragali, Radix Glycyrrhizae alcohol extraction residue is got in the preparation of need testing solution, filters, and precision is measured subsequent filtrate 1ml, puts in the 100ml measuring bottle, and thin up shakes up to scale, promptly gets;
The algoscopy precision is got need testing solution 1.0ml; Put in the 10ml tool plug test tube, the sighting target directrix curve prepares method down from " precision adding sulphuric acid anthrone solution 4ml...... in ice bath ", measures trap in accordance with the law; Read the content of total polysaccharides the need testing solution from standard curve, promptly get;
The content assaying method of c, cinobufagin, bufogenin:
Measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D);
Chromatographic condition and system suitability test use octadecylsilane chemically bonded silica to be filler; 0.5% potassium dihydrogen phosphate-acetonitrile 50: 50 is regulated pH 3.2 with phosphoric acid; The detection wavelength is 296nm; 40 ℃ of column temperatures; Number of theoretical plate calculates by cinobufagin peak, bufogenin peak and is not less than 4000 respectively;
The cinobufagin reference substance is got in the preparation of reference substance solution, the bufogenin reference substance is an amount of, and accurate the title decides, and adds methanol and processes the solution that 1ml respectively contains cinobufagin, bufogenin 50ug respectively, promptly gets;
Each batch Venenum Bufonis is got in the preparation of need testing solution respectively, the Mylabris decocting boils the concentrated solution semi-finished product; Precision is measured 20.0ml, puts in the separatory funnel, with extracted with diethyl ether 3 times; Each 30ml; Merge ether solution, use a small amount of anhydrous sodium sulfate dehydration, concentrating under reduced pressure reclaims ether; Water bath method; Residue, shakes up to the 5ml measuring bottle with methanol constant volume, promptly gets; Face with preceding 0.45ul membrane filtration;
Accurate respectively above-mentioned reference substance solution and each 10ul of need testing solution of drawing of algoscopy injects high performance liquid chromatograph, measures, and promptly gets;
The content assaying method of d, cantharidin:
Measure according to liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D);
Chromatographic condition and system suitability test use octadecylsilane chemically bonded silica to be filler; With acetonitrile-water is mobile phase at 80: 20; The detection wavelength is 209nm; Number of theoretical plate calculates by the cantharidin peak and is not less than 1500;
It is an amount of that the cantharidin reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methanol and processes the solution that every 1ml contains 0.5mg, promptly gets;
Each batch Venenum Bufonis is got in the preparation of need testing solution respectively, the Mylabris decocting boils the concentrated solution semi-finished product; Precision is measured 20.0ml, puts in the separatory funnel, with water saturation chloroform extraction 3 times; Each 30ml; Combined chloroform liquid is used a small amount of anhydrous sodium sulfate dehydration, and concentrating under reduced pressure reclaims chloroform; Water bath method; Residue, shakes up to the 2ml measuring bottle with methanol constant volume, promptly gets; Face with preceding 0.45ul membrane filtration;
Accurate respectively reference substance solution and each 10ul of need testing solution of drawing of algoscopy injects chromatograph of liquid, calculates, and promptly gets;
Water solublity indoles alkaloid content assaying method in e, the Venenum Bufonis:
Measure according to spectrophotometer method (appendix V of Chinese Pharmacopoeia version in 2000);
The preparation precision of reference substance solution takes by weighing puts that the 5-hydroxyl color behind the vacuum drying 24h adds water and processes every milliliter of solution that contains 33ug by reference substance in the phosphorus pentoxide desiccator, promptly gets;
Each batch Mylabris is got in the preparation of need testing solution respectively, the Venenum Bufonis decocting boils the concentrated solution semi-finished product, and precision is measured 10.0ml, adds water to 100ml, promptly gets;
The accurate need testing solution 0.5ml that draws of algoscopy puts the 10ml volumetric flask, adds water and makes into 5ml; The accurate 15% paradime thylaminobenzaldehyde hydrochloric acid solution 5ml that adds shakes up, and room temperature was placed 30 minutes, and water is cooked blank, measures trap at wavelength 555nm place, from the amount that standard curve is read indoles total alkaloids the sample solution, calculates content.
In Chinese medicine oral liquid of the present invention, Radix Ginseng Rubra, Radix Astragali strongly invigorating primordial QI, strengthening the body resistance, invigorating the spleen and benefiting QI; Astragalus polysaccharides more helps the detumescence effect, can prevent that hepatic glycogen from reducing, and improves mononuclear phagocyte and leukocytic phagocytic activity, raise immunity; Radix Ginseng more can cause the death of cancerous cell.Mylabris hard masses softening and resolving, activating blood and removing stasis, the medicament that extracts through above-mentioned technology contains a large amount of cantharidin, is used for treating various tumor patients; Therefore bufogenin has the effect that pain relieving is had one's ideas straightened out, and is Venenum Bufonis removing blood stasis eliminating stagnation, removes the material base of effects such as the pain over the hypochondriac region, and obvious enhanced NK cell activity has the obvious suppression effect to tumor.The present invention is used for advanced primary liver cancer qi deficiency and Stagnation card, and disease is seen right latus palpatable abdominal mass, unmovable pain, abdominal distention and little diet, fatigue and weakness etc.
Animal test results prompting before clinical: Chinese medicine oral liquid of the present invention promotes that cancerous cell breaks up and cancer cell-apoptosis again; The external equal growth that can suppress multiple cancerous cell in vivo; To chemotherapy, the insensitive gland type of radiotherapy cancerous cell, very strong inhibitory action is arranged also; Cancerous tissue cAMP/cGMP ratio is strengthened, improve immunologic function, effectively remove interior free yl.
The present invention compares with the prior art DELISHENG ZHUSHEYE:
(1) DELISHENG ZHUSHEYE is owing to be injection, and therefore thoroughly necessary to removing of impurity in the injection and thermal source, its process using calcium hydroxide, sulfur dioxide are removed impurity.The medicinal liquid adding calcium hydroxide could effectively be removed thermal source to pH 12 in strong alkaline aqueous solution, cause the loss amount of effective ingredient total saponins, total polysaccharides also very big.Oral liquid of the present invention needn't be considered thermal source; Ginsenoside, Radix Astragali saponin are with ethanol extraction; Need not further to remove impurity and thermal source with the strong basicity calcium hydroxide, only polysaccharide extraction liquid is removed impurity with micro-calcium hydroxide pH 8.0, so the extraction purification loss amount of effective ingredient is few.The extracted amount 15% of DELISHENG ZHUSHEYE polysaccharide, loss amount reaches 85%; The extracted amount of Radix Ginseng, Radix Astragali total saponins only 20%, loss amount can reach 80%.And the Radix Ginseng of oral liquid of the present invention, Radix Astragali total saponins, total polysaccharides can reach more than 85% after extracting purification, loss amount only 15%;
(2) Venenum Bufonis in the DELISHENG ZHUSHEYE is only used ethanol extraction liposoluble part; Water-soluble part discards; And the Venenum Bufonis alkaloid has better antitumaous effect in the water-soluble part of Venenum Bufonis, and at present to sell cinobufacin oral liquid, HUACHANSU ZHUSHEYE all be to be main anticancer effective component with water-soluble part Venenum Bufonis alkaloid in the market.Cause medicinal liquid unstable because water-soluble part exists with the liposoluble part simultaneously in the DELISHENG ZHUSHEYE, therefore can only discard water-soluble part;
(3) Radix Glycyrrhizae is very important simply medicine in the raw material; DELISHENG ZHUSHEYE compatibility Radix Glycyrrhizae can cause medicinal liquid unstable, promptly produces deposition after the sterilization, and sterilization back pH reduces, and glycyrrhizic acid is lower than at 5.5 o'clock and promptly precipitates at pH separates out; And glycyrrhizic acid has antitumaous effect; Can strengthen the antitumaous effect of Radix Ginseng, the Radix Astragali, Venenum Bufonis, Mylabris; Radix Glycyrrhizae is separated hundred toxic actions simultaneously; The dysurea that the compatibility Radix Glycyrrhizae causes Mylabris, frequent micturition, urgent micturition can obviously reduce; Can disappear basically to gastrointestinal side effect, the mouse toxicity experiment shows oral liquid of the present invention if do not add the mice LD of Radix Glycyrrhizae
50Be 54.25g/kg, add the LD behind the Radix Glycyrrhizae
50Be 61.86g/kg, show that toxicity obviously reduces.
The specific embodiment
Embodiment 1
One, raw material sources
1, Radix Ginseng Rubra: these article derive from ginseng field, Fujin, Heilongjiang Province county, purchase the Chinese crude drug company in the Leshan.Through identifying cultivation article (practise claim " Park Ginseng ") the dry root and rhizome through steam after of these article for Araliaceae Radix Ginseng Rubra Panax ginseng C.A.Mey..Meet " the pertinent regulations under the 121st page of Radix Ginseng Rubra item of Chinese pharmacopoeia version in 2000.
2, the Radix Astragali: through identifying the dry root of these article for leguminous plant Radix Astagali Astragalusmembranaceus (Fisch) Bge.Var.mongholicus (Bge.) Hsiao or Radix Astragali Astragalus membranaceus (Fisch) Bge...Meet " the pertinent regulations under the 249th page of Radix Astragali item of Chinese pharmacopoeia version in 2000.
3, Radix Glycyrrhizae: through identifying the dry root and rhizome of these article for glycyrrhizic legume Glycyrrhiza uralensis Fisch Glycyrrhiza inflata Bat. Glycyhisa inflata.Bat or Glycyrrhiza glabra L. Glycyrrhiza glabru L..Meet " the pertinent regulations under the 59th page of Radix Glycyrrhizae item of Chinese pharmacopoeia version in 2000.
4, Mylabris: through identifying the dry body of these article for Meloidae insecticide south mylabris phalerata Mylabris phaleratapallas or yellow black mylabris cichorii Mylabris cichorii Linnaeus.Meet " the pertinent regulations under the 273rd page of Mylabris item of Chinese pharmacopoeia version in 2000.
5, Venenum Bufonis: through being accredited as the dry secretions of Bufonidae animal Bufo siccus Bufo bufo gargarizansCantor or black surround Bufo siccus Bufo melanostictus Schneider.Meet " the pertinent regulations under the 316th page of Venenum Bufonis item of Chinese pharmacopoeia version in 2000.
Two, the pre-treatment of medical material and processing
1, Radix Ginseng Rubra cleans, selects, removes impurity, dries, cuts into slices, pulverizes 40 mesh sieves, weighs.
2, the Radix Astragali cleans, selects, removes impurity, dries, cuts into slices, pulverizes 40 mesh sieves, weighs.
3, impurity is cleaned, selects, removed to Radix Glycyrrhizae, dries, cuts into slices, pulverized 40 mesh sieves, weighs.
4, Mylabris selects, removes impurity, pulverizes 20 mesh sieves, weighs.
5, impurity is selected, removed to Venenum Bufonis, pulverized 40 mesh sieves, weighs.
Three, preparation technology
1, gets Radix Ginseng Rubra 120 grams, the Radix Astragali 690 grams, Radix Glycyrrhizae 192 grams, Mylabris 2 grams, Venenum Bufonis 1 gram.
2, Radix Ginseng Rubra, the Radix Astragali, Radix Glycyrrhizae being pulverized is coarse powder, adds 8 times of amount 75% ethanol, reflux, extract, 3 times, and each 1.5 hours, merge alcohol extract, filter, filtrate recycling ethanol to nothing alcohol flavor obtains extracting solution A;
Add 8 times of water gagings to residue and decoct 3 times, each 20 minutes, merge decocting liquid; Add the calcium hydroxide breast and regulate pH 8.0, add 10% sodium hydroxide solution and regulate pH 9.0, filter; Filtrating is regulated pH 6.5-7.0 with dilute hydrochloric acid, and filtrate decompression concentrates, and obtains extracting solution B.
3, Mylabris being pulverized is coarse powder, and Venenum Bufonis is pulverized and is to mix fine powder, adds 7 times of amount 95% ethanol, reflux, extract, 3 times, and each 20 minutes, the merging alcohol extract, concentrating under reduced pressure obtains extracting solution C;
Add 7 times of water gagings to residue and decoct 3 times, each 20 minutes, merge decocting liquid, left standstill 2 hours, to filter, filtrate decompression concentrates, and obtains extracting solution D.
4, above extracting solution A, B, C, D are mixed, stir, the embedding of adjustment volume is in infusion bottle, and 100 ℃ of sterilizations 30 minutes were left standstill more than 1 week, filtered, and added the sucrose mixing to filtrating, thin up to 2 times, and embedding is in oral liquid bottle, and sterilization promptly gets.
Embodiment 2
Adopt Radix Ginseng Rubra 60 grams, the Radix Astragali 345 grams, Radix Glycyrrhizae 96 grams, Mylabris 1 gram, Venenum Bufonis 0.5 gram, surplus with embodiment 1.
Embodiment 3
Adopt Radix Ginseng Rubra 240 grams, the Radix Astragali 1380 grams, Radix Glycyrrhizae 384 grams, Mylabris 4 grams, Venenum Bufonis 2 grams, surplus with embodiment 1.
Claims (7)
1. Chinese medicine oral liquid of treating cancer, its effective ingredient is processed by Radix Ginseng Rubra 60-240 weight portion, Radix Astragali 345-1380 weight portion, Radix Glycyrrhizae 96-384 weight portion, Mylabris 1-4 weight portion, Venenum Bufonis 0.5-2 weight portion.
2. according to the Chinese medicine oral liquid of claim 1, its effective ingredient is processed by Radix Ginseng Rubra 120 weight portions, the Radix Astragali 690 weight portions, Radix Glycyrrhizae 192 weight portions, Mylabris 2 weight portions, Venenum Bufonis 1 weight portion.
3. the method for preparing of Chinese medicine oral liquid according to claim 1 comprises Radix Ginseng Rubra, the Radix Astragali, Radix Glycyrrhizae, Mylabris, Venenum Bufonis is used earlier alcohol reflux, residue reuse water boiling and extraction.
4. according to the method for preparing of claim 3, comprise Radix Ginseng Rubra, the Radix Astragali, Radix Glycyrrhizae are pulverized, add 75% alcohol reflux, obtain extracting solution A; The residue decocte with water is extracted, and adds diluted alkaline to extracting solution and regulates pH＞8, filters, and filtrating is regulated pH 6.5-7.0 with diluted acid, and concentrating under reduced pressure obtains extracting solution B;
Mylabris, Venenum Bufonis are pulverized, added 95% alcohol reflux, obtain extracting solution C; The residue decocte with water is extracted, and filters, and filtrate decompression concentrates, and obtains extracting solution D;
With above extracting solution A, B, C, D mix homogeneously, process oral liquid.
5. according to the method for preparing of claim 4, comprise that it is coarse powder that Radix Ginseng Rubra, the Radix Astragali, Radix Glycyrrhizae are pulverized, add 8 times of amount 75% ethanol, reflux, extract, 3 times, each 1.5 hours, merge alcohol extract, filter, filtrate recycling ethanol to nothing alcohol flavor obtains extracting solution A; Add 8 times of water gagings to residue and decoct 3 times, each 20 minutes, merge decocting liquid; Add the calcium hydroxide breast and regulate pH 8.0, add 10% sodium hydroxide solution and regulate pH 9.0, filter; Filtrating is regulated pH 6.5-7.0 with dilute hydrochloric acid, and filtrate decompression concentrates, and obtains extracting solution B;
It is coarse powder that Mylabris is pulverized, and Venenum Bufonis is pulverized and is to mix fine powder, adds 7 times of amount 95% ethanol, reflux, extract, 3 times, and each 20 minutes, the merging alcohol extract, concentrating under reduced pressure obtains extracting solution C; Add 7 times of water gagings to residue and decoct 3 times, each 20 minutes, merge decocting liquid, left standstill 2 hours, to filter, filtrate decompression concentrates, and obtains extracting solution D;
Above extracting solution A, B, C, D are mixed, stir, the embedding of adjustment volume is in infusion bottle, and 100 ℃ of sterilizations 30 minutes were left standstill more than 1 week, filtered, and added the sucrose mixing to filtrating, thin up to 2 times, and embedding is in oral liquid bottle, and sterilization promptly gets.
6. the active constituent content measuring method of Chinese medicine oral liquid according to claim 1 comprises the content assaying method of total saponins, polysaccharide, cinobufagin and bufogenin, cantharidin, water solublity indoles.
7. according to the content assaying method of claim 6, comprising:
The content assaying method of a, total saponins:
Measure according to ultraviolet spectrophotometry (2000 editions one appendix V A of Chinese Pharmacopoeia);
The preparation precision of reference substance solution is got red ginseng saponin Re reference substance 8.0mg, and accurate the title decides, and puts in the 5ml volumetric flask, adds methanol and makes dissolving in right amount, and be diluted to scale, shakes up promptly to get, and every milliliter contains red ginseng saponin Re 1.6mg;
The preparation precision of standard curve is measured reference substance solution 20,40,60,80,100ul, puts respectively in the 10ml tool plug test tube, puts and waves most solvent in the water-bath, takes out cooling immediately; The accurate glacial acetic acid solution 0.2ml that adds 5% vanillin, perchloric acid 0.8ml shakes up, and puts heating taking-up in 15 minutes in 60 ℃ of water-baths, immediately with circulating water cooling 2 minutes; The accurate again glacial acetic acid solution 5ml that adds shakes up, and is blank with the corresponding reagent; According to 33 pages of appendix of spectrophotography Pharmacopoeia of the People's Republic of China version in 2000; Wavelength at 544nm is measured trap, is vertical coordinate with the trap, and concentration is the standard curve that abscissa is drawn solution;
Accurate Radix Ginseng Rubra, the Radix Astragali, the Radix Glycyrrhizae semi-finished product concentrated solution 0.5ml of drawing of the preparation of need testing solution puts in the 100ml measuring bottle, and thin up is to scale, close plug, shakes up; The above-mentioned solution 10ml of accurate again absorption; Put in the separatory funnel, with water saturation n-butanol extraction three times, each 20ml; Merge butanol extraction liquid; With washing twice, each 10ml discards water liquid; Evaporate to dryness in the water-bath of n-butyl alcohol liquid; Residue is put in the 5ml measuring bottle with dissolve with methanol, and is diluted to scale, shakes up, promptly gets;
The accurate need testing solution 40ul that draws of algoscopy; Put in the 10ml tool plug test tube; Put and wave most solvent in the water-bath; Take out cooling immediately; The sighting target directrix curve prepares method down from " the vanillin glacial acetic acid solution 0.2ml...... of precision adding 5% ", measures trap in accordance with the law, reads the content of Radix Ginseng Rubra total saponins the need testing solution from standard curve; Calculate, promptly get;
B, content Determination of Polysaccharide method
Measure according to spectrophotography (appendix V of Chinese Pharmacopoeia version in 2000);
The preparation precision of reference substance solution takes by weighing the anhydrous glucose 80mg that is dried to constant weight through 80 ℃, puts in the 100ml volumetric flask, is dissolved in water, and is diluted to scale; Shake up, precision is measured 5ml, puts in the 50ml volumetric flask; Thin up is to scale, shakes up promptly to get, and per 1 milliliter contains anhydrous glucose 0.08mg;
The preparation precision of standard curve is measured reference substance solution 0.2,0.4,0.6,0.8,1.0ml; Put respectively in the 10ml tool plug test tube; Each is managed precision and adds water and make into 1ml; In ice bath, add sulphuric acid anthrone solution 4ml respectively; Shake up; Put in the boiling water bath and heated 10 minutes; Take out; After placing room temperature; With the corresponding reagent is blank, according to appendix of spectrophotography Pharmacopoeia of the People's Republic of China version in 2000, measures trap in the wavelength of 625nm; With the trap is vertical coordinate, and concentration is the standard curve that abscissa is drawn solution; Said sulphuric acid anthrone solution is like this preparation: get anthrone 50mg, add water 25ml and sulphuric acid 75ml makes dissolving, promptly get, face and use stylish preparation;
The water decoction of Radix Ginseng Rubra, the Radix Astragali, Radix Glycyrrhizae alcohol extraction residue is got in the preparation of need testing solution, filters, and precision is measured subsequent filtrate 1ml, puts in the 100ml measuring bottle, and thin up shakes up to scale, promptly gets;
The algoscopy precision is got need testing solution 1.0ml; Put in the 10ml tool plug test tube, the sighting target directrix curve prepares method down from " precision adding sulphuric acid anthrone solution 4ml...... in ice bath ", measures trap in accordance with the law; Read the content of total polysaccharides the need testing solution from standard curve, promptly get;
The content assaying method of c, cinobufagin, bufogenin:
Measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D);
Chromatographic condition and system suitability test use octadecylsilane chemically bonded silica to be filler; 0.5% potassium dihydrogen phosphate-acetonitrile 50: 50 is regulated pH 3.2 with phosphoric acid; The detection wavelength is 296nm; 40 ℃ of column temperatures; Number of theoretical plate calculates by cinobufagin peak, bufogenin peak and is not less than 4000 respectively;
The cinobufagin reference substance is got in the preparation of reference substance solution, the bufogenin reference substance is an amount of, and accurate the title decides, and adds methanol and processes the solution that 1ml respectively contains cinobufagin, bufogenin 50ug respectively, promptly gets;
Each batch Venenum Bufonis is got in the preparation of need testing solution respectively, the Mylabris decocting boils the concentrated solution semi-finished product; Precision is measured 20.0ml, puts in the separatory funnel, with extracted with diethyl ether 3 times; Each 30ml; Merge ether solution, use a small amount of anhydrous sodium sulfate dehydration, concentrating under reduced pressure reclaims ether; Water bath method; Residue, shakes up to the 5ml measuring bottle with methanol constant volume, promptly gets; Face with preceding 0.45ul membrane filtration;
Accurate respectively above-mentioned reference substance solution and each 10ul of need testing solution of drawing of algoscopy injects high performance liquid chromatograph, measures, and promptly gets;
The content assaying method of d, cantharidin:
Measure according to liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D);
Chromatographic condition and system suitability test use octadecylsilane chemically bonded silica to be filler; With acetonitrile-water is mobile phase at 80: 20; The detection wavelength is 209nm; Number of theoretical plate calculates by the cantharidin peak and is not less than 1500;
It is an amount of that the cantharidin reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methanol and processes the solution that every 1ml contains 0.5mg, promptly gets;
Each batch Venenum Bufonis is got in the preparation of need testing solution respectively, the Mylabris decocting boils the concentrated solution semi-finished product; Precision is measured 20.0ml, puts in the separatory funnel, with water saturation chloroform extraction 3 times; Each 30ml; Combined chloroform liquid is used a small amount of anhydrous sodium sulfate dehydration, and concentrating under reduced pressure reclaims chloroform; Water bath method; Residue, shakes up to the 2ml measuring bottle with methanol constant volume, promptly gets; Face with preceding 0.45ul membrane filtration;
Accurate respectively reference substance solution and each 10ul of need testing solution of drawing of algoscopy injects chromatograph of liquid, calculates, and promptly gets;
Water solublity indoles alkaloid content assaying method in e, the Venenum Bufonis:
Measure according to spectrophotometer method (appendix V of Chinese Pharmacopoeia version in 2000);
The preparation precision of reference substance solution takes by weighing puts that the 5-hydroxyl color behind the vacuum drying 24h adds water and processes every milliliter of solution that contains 33ug by reference substance in the phosphorus pentoxide desiccator, promptly gets;
Each batch Mylabris is got in the preparation of need testing solution respectively, the Venenum Bufonis decocting boils the concentrated solution semi-finished product, and precision is measured 10.0ml, adds water to 100ml, promptly gets;
The accurate need testing solution 0.5ml that draws of algoscopy puts the 10ml volumetric flask, adds water and makes into 5ml; The accurate 15% paradime thylaminobenzaldehyde hydrochloric acid solution 5ml that adds shakes up, and room temperature was placed 30 minutes, and water is cooked blank, measures trap at wavelength 555nm place, from the amount that standard curve is read indoles total alkaloids the sample solution, calculates content.
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| CN103278466A (en) * | 2013-05-02 | 2013-09-04 | 广西北斗星动物保健品有限公司 | Quality control method for toad venom injection |
| US20150313835A1 (en) * | 2014-05-05 | 2015-11-05 | Access Business Group International Llc | Compositions including a botanical extract and methods of using the same in skin whitening and skin lightening applications |
| US9801809B2 (en) * | 2014-05-05 | 2017-10-31 | Access Business Group International Llc | Compositions including a botanical extract and methods of using the same in skin whitening and skin lightening applications |
| CN110314175A (en) * | 2019-04-18 | 2019-10-11 | 张家口邦合生物科技有限公司 | A kind of antitumor traditional Chinese medicinal components and preparation method thereof comprising white ganoderma lucidum |
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