CN1943642A - Preparing method for a kind of medicinal composition and its effective parts and active ingredients - Google Patents

Preparing method for a kind of medicinal composition and its effective parts and active ingredients Download PDF

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CN1943642A
CN1943642A CN 200610113835 CN200610113835A CN1943642A CN 1943642 A CN1943642 A CN 1943642A CN 200610113835 CN200610113835 CN 200610113835 CN 200610113835 A CN200610113835 A CN 200610113835A CN 1943642 A CN1943642 A CN 1943642A
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parts
silica gel
chromatographic isolation
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stream part
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CN1943642B (en
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石任兵
王永炎
刘斌
徐秋萍
王彦志
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Beijing University of Chinese Medicine
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Beijing University of Chinese Medicine
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Abstract

This invention provides a kind of medicinal composition and its effective parts and active ingredients. The medicinal composition in this invention is composed with ginseng, puerarin,radices paeoniae rubra. Preparation of the effective part of the medicinal composition in this invention is: weigh the raw medical materials according there requirements, mix the up evenly, distill by adding ethanol for several times, put each of the distilled liquid together, condense the liquid through decompression, add water into the condensed liquid and stir evenly, then adsorb it by using large meshed adsorptive resin, wash the resin mesh until the wash water is with no color, then use ethanol to wash it, collect the ethanol washing liquid, collect back solvent and make it dry by decompression. After all these steps, the effective parts of the medicinal composition of this invention are achieved. The medicinal composition and its effective parts or active ingredients can be used in preparing the medicine for treatment of dementia due to blood vessel problem.

Description

The preparation method of a kind of pharmaceutical composition and effective site thereof, active component
Technical field
The present invention relates to the preparation method of a kind of pharmaceutical composition and effective site thereof, active component, particularly the preparation method of a kind of pharmaceutical composition of anti-angiogenic dementia and effective site thereof, active component.
Background technology
Vascular dementia (VD) be occur on the cerebrovascular basis based on memory, cognitive function is damaged, or with the acquired intelligent persistence infringement of language, visual space ability and emotion, personality disorder, hypophrenias such as memory, orientation force, computing power, faculty of understanding appearred in the patient after it mainly showed as cerebrovascular, can influence patient's life and social activity with linguistic competence's decline, emotion and personality change etc.Its sickness rate constantly raises, and has become the commonly encountered diseases and the frequently-occurring disease that influence middle-aged and elderly people Health and Living quality.Along with the development of China's aged tendency of population, bring white elephant for society and family.According to investigations, the sickness rate of over-65s dementia is 5%.U.S.'s statistics, 26.3% meeting generation dementia among the ischemic cerebrovascular survival patient more than 60 years old.At present, doctor trained in Western medicine is to the treatment of VD, and the one, prevention and treatment cerebrovascular, the 2nd, improve brain function, alleviate dementia symptom.Chinese medicine and pharmacy thinks that then " turbid poison damage brain network " is the main pathogenesis of VD, and the kidney essense deficiency of vital energy, phlegm and blood stasis, obstruction of collateral are the bases that VD takes place, and that the expectorant stasis of blood is accumulate is long-pending, make and give birth to turbid poison, ruins brain network brains, is to cause VD that the key of development takes place.Facts have proved, Chinese medicine is delaying the state of an illness, is improving General Symptoms, is improving to a certain extent and have advantage aspect intelligent, to improve memory, orientation force has certain effect, especially improve the life quality aspect outstanding role arranged improving patient's dull expression, subjective symptoms and initiative and improvement.But from pure Chinese medicinal preparation, isolate method and related preparations at present and do not seen also that report was arranged with the active drug effective region of anti-angiogenic dementia and active component.
Summary of the invention
The object of the present invention is to provide a kind of pharmaceutical composition; Another object of the present invention is to provide a kind of pharmaceutical composition of anti-angiogenic dementia; The 3rd purpose of the present invention is to provide the preparation method of this pharmaceutical composition effective site, active component; The 4th purpose of the present invention is to provide the new pharmaceutical use of this pharmaceutical composition and effective site thereof.
The objective of the invention is to be achieved through the following technical solutions:
The raw material of pharmaceutical composition of the present invention consists of:
Radix Ginseng 3～18 weight portions, Radix Puerariae 3～18 weight portions, Radix Paeoniae Rubra 3～18 weight portions.
The raw material composition of pharmaceutical composition of the present invention is preferably:
Radix Ginseng 10 weight portions, Radix Puerariae 10 weight portions, Radix Paeoniae Rubra 10 weight portions.
The raw material composition of pharmaceutical composition of the present invention is preferably:
Radix Ginseng 3 weight portions, Radix Puerariae 18 weight portions, Radix Paeoniae Rubra 9 weight portions.
The raw material composition of pharmaceutical composition of the present invention is preferably:
Radix Ginseng 18 weight portions, Radix Puerariae 3 weight portions, Radix Paeoniae Rubra 12 weight portions.
The above-mentioned raw materials medicine adds conventional adjuvant, according to common process, makes the dosage form of clinical acceptance, includes but not limited to capsule, pill, tablet, granule, oral liquid or injection.
The preparation method of pharmaceutical composition effective site of the present invention is optional following a kind of:
A, weighting raw materials according to the above ratio, mix homogeneously, 30～80% ethanol extractions 1～4 time that add 6～10 times of parts by volume, each 1～3 hour, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, be evaporated to 1-3 times of volume of medical material amount under the condition that temperature is 70～80 ℃, relative density is about 1.04～1.05 concentrated solution under 60～65 ℃ the condition; Get concentrated solution, the water that adds 2-6 times of medical material amount stirs, adsorb by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery promptly gets pharmaceutical composition effective site of the present invention to doing;
B, take by weighing Radix Ginseng, 30～80% ethanol extractions 1～4 time that add 6～10 times of parts by volume, each 1～3 hour, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, is evaporated to 1-3 times of volume of medical material amount under the condition that temperature is 70～80 ℃, and relative density is about 1.04～1.05 concentrated solution under 60～65 ℃ the condition; Get concentrated solution, the water that adds 2-6 times of medical material amount stirs, adsorb by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery gets the Radix Ginseng extract part to doing;
Take by weighing Radix Puerariae, 30～80% ethanol extractions 1～4 time that add 6～10 times of parts by volume, each 1～3 hour, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, is evaporated to 1-3 times of volume of medical material amount under 70～80 ℃ of conditions of temperature, and relative density is about 1.04～1.05 concentrated solution under 60～65 ℃ of conditions; Get concentrated solution, the water that adds 2-6 times of medical material amount stirs, adsorb by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery gets the Radix Puerariae extract part to doing;
Take by weighing Radix Paeoniae Rubra, 30～80% ethanol extractions 1～4 time that add 6～10 times of parts by volume, each 1～3 hour, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, is evaporated to 1-3 times of volume of medical material amount under 70～80 ℃ of conditions of temperature, and 60～65 ℃ of following relative densities are about 1.04～1.05 concentrated solution; Get concentrated solution, the water that adds 2-6 times of medical material amount stirs, adsorb by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery gets the Radix Paeoniae Rubra extract part to doing;
Get above-mentioned Radix Ginseng extract part, Radix Puerariae extract part and Radix Paeoniae Rubra extract part, by 1～20: 1～20: 1～20 mixed is even, promptly gets pharmaceutical composition effective site of the present invention.
The preparation method of pharmaceutical composition effective site of the present invention is preferably as follows a kind of:
A, weighting raw materials according to the above ratio, mix homogeneously, 30～80% ethanol extractions 3 times that add 8 times of parts by volume, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, be evaporated to 2 times of volumes of medical material amount under 70～80 ℃ of conditions of temperature, relative density is about 1.04～1.05 concentrated solution under 60～65 ℃ of conditions; Get concentrated solution, the water that adds 4 times of medical material amounts stirs, adsorb by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery promptly gets pharmaceutical composition effective site of the present invention to doing;
B, take by weighing Radix Ginseng, 30～80% ethanol extractions 3 times that add 8 times of parts by volume, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, is evaporated to 2 times of volumes of medical material amount under the condition that temperature is 70～80 ℃, and relative density is about 1.04～1.05 concentrated solution under 60～65 ℃ the condition; Get concentrated solution, the water that adds 4 times of medical material amounts stirs, adsorb by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery gets the Radix Ginseng extract part to doing;
Take by weighing Radix Puerariae, 30～80% ethanol extractions 3 times that add 8 times of parts by volume, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, is evaporated to 2 times of volumes of medical material amount under 70～80 ℃ of conditions of temperature, and relative density is about 1.04～1.05 concentrated solution under 60～65 ℃ of conditions; Get concentrated solution, the water that adds 4 times of medical material amounts stirs, adsorb by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery gets the Radix Puerariae extract part to doing;
Take by weighing Radix Paeoniae Rubra, 30～80% ethanol extractions 3 times that add 8 times of parts by volume, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, is evaporated to 2 times of volumes of medical material amount under 70～80 ℃ of conditions of temperature, and 60～65 ℃ of following relative densities are about 1.04～1.05 concentrated solution; Get concentrated solution, the water that adds 4 times of medical material amounts stirs, adsorb by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery gets the Radix Paeoniae Rubra extract part to doing;
Get above-mentioned Radix Ginseng extract part, Radix Puerariae extract part and Radix Paeoniae Rubra extract part, by 1: 1: 1,1: 20: 1,1: 1: 20,1: 10: 10 or 3: 4: 3 mixed were even, promptly get pharmaceutical composition effective site of the present invention;
Pharmaceutical composition effective site of the present invention is light brown powder, has certain moisture, and soluble in water and Diluted Alcohol is insoluble in lipotropy organic solvents such as chloroform, petroleum ether; The ferric chloride reaction positive, aluminum chloride reacting positive, hydrochloric acid-magnesium powder reaction negative, the acetic anhydride-strong sulfuric acid response positive, the alpha-Naphthol-strong sulfuric acid response positive; Mainly contain osajin, monoterpene glycosides and saponin component, wherein puerarin content is 15.0～25.0%, daiazi, 3 '-total content of methoxy puerarin and celery glycosyl puerarin is 12.0～18.0%, paeoniflorin content is 7.0～15.0%, and the content of Radix Ginseng total saponins is 17.0～25.0%.
Pharmaceutical composition effective site of the present invention adds conventional adjuvant, according to common process, makes the dosage form of clinical acceptance, includes but not limited to capsule, pill, tablet, granule, oral liquid or injection.
The preparation method of pharmaceutical composition active component of the present invention is:
The separation of effective ingredient in the Radix Paeoniae Rubra: get Radix Paeoniae Rubra 3-18 weight portion, with 30～80% ethanol extractions of 6-10 times of parts by volume 1-4 time, each 1-3 hour, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, after being evaporated to the 1500-2500 parts by volume under 70～80 ℃ of conditions of temperature, be diluted with water to the 4000-6000 parts by volume, by AB-8 type macroporous adsorptive resins, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, and collect ethanol elution, decompression and solvent recovery is to doing, residue is put 50～70 ℃ of dryings in the vacuum drying oven, obtains dry extract;
Get the dry extract 400-600 weight portion of method for preparing, carry out silica gel column chromatography and separate (chromatographic isolation I), with 100: 0 → 20: 1 → 10: 1 → 5: 1 → 2: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 115～125 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part the 4th～6 that chromatographic isolation I is obtained, 7～16,17～31,32～41,42～74,75～99,100～121 merge respectively, obtain CA, CB, CC, CD, CE, CF, seven different pieces of CG;
Portion C A carries out silica gel column chromatography to be separated, and promptly chromatographic isolation CAI uses 2: 1 → 7: 5 gradient elutions of petroleum ether-acetone mixed solvent, portioning is collected, every 40-60 parts by volume collection is a, collects altogether to obtain 20～25 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part; Stream part 5～17 that chromatographic isolation CAI obtains merges, silica gel column chromatography separates, and promptly chromatographic isolation CAII uses petroleum ether-acetone mixed solvent 40～60: 1 eluting, portioning is collected, every 40-60 parts by volume collection is a, collects altogether to obtain 8～12 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 3～5 that chromatographic isolation CAII obtains merges, separate out the white plates crystallization, use the petroleum ether recrystallization, obtain benzoic acid 0.01-0.03 weight portion;
Portion C B carries out silica gel column chromatography to be separated, be chromatographic isolation CBI, with 100: 0 → 50: 1 → 20: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 40-60 parts by volume collection is a, collects altogether to obtain 30～35 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 22～27 that chromatographic isolation CBI obtains merges, separate through silica gel column chromatography, promptly chromatographic isolation CBII uses petroleum ether-acetone mixed solvent (7: 2) eluting, portioning is collected, every 50ml collection is a, collects altogether to obtain 10～15 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 9 that chromatographic isolation CBII obtains is separated through Sephadex LH-20 column chromatography, methanol-eluted fractions obtains 1S, 2S, 4R-is trans-2-hydroxyl-1, and 8-cineole 0.004-0.006 weight portion and cupreol 0.01-0.03 weight portion;
Portion C D carries out silica gel column chromatography to be separated, be chromatographic isolation CDI, use 2-4: the petroleum ether-acetone of 1 ratio → 5-10: the petroleum ether-acetone of 5 ratios → methanol gradient elution, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 75～80 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 41～70 that chromatographic isolation CDI obtains merges, and separates with silica gel column chromatography, be chromatographic isolation CDII, the petroleum ether-ethyl acetate mixed solvent eluting of 2: 1 ratios, portioning is collected, and every 40-60 parts by volume is collected a, collect altogether and obtain 25～30 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 9～16 that chromatographic isolation CDII obtains merges, and separates through silica gel column chromatography, 4-6: the petroleum ether of 1 ratio-acetone eluting obtains dihydro apigenin 0.006-0.01 weight portion; The methanol-eluted fractions thing that chromatographic isolation CDI obtains separates with silica gel column chromatography, be chromatographic isolation CDIII, 2-3: the petroleum ether of 1 ratio-acetone mixed solvent eluting, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 25～30 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 9～18 that chromatographic isolation CDIII obtains merges, separate with Sephadex LH-20 column chromatography, be chromatographic isolation CDIV, the acetone eluting, portioning is collected, every 20-40 parts by volume is collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 5～12 that chromatographic isolation CDIV obtains merges, separate through silica gel column chromatography, be chromatographic isolation CDV, 50: 1 → 20: 1 → 10: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 45～50 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 8～12 that chromatographic isolation CDV obtains merges, and separates out white crystals, obtains lacdtlorin 0.02-0.04 weight portion with acetone recrystallization, stream part 13～30 merging, separate i.e. chromatographic isolation CDVI, 20: 1 → 10: 1 gradient elutions of chloroform-methanol mixed solvent through silica gel column chromatography, portioning is collected, every 40-60 parts by volume collection is a, collects altogether to obtain 35～40 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 4～18 that chromatographic isolation CDVI obtains merges, through the preparation of preparation of silica gel chromatographic isolation, 20-40: the chloroform-methanol mixed solvent of 1 ratio launches, and obtains benzoylpaeoniflorin 0.03-0.05 weight portion; Stream part 25～36 merging, separate (chromatographic isolation CDVII) through Sephadex LH-20 column chromatography, the acetone eluting, portioning is collected, and every 20ml collects a, collect altogether and obtain 10～12 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 1～3 that chromatographic isolation CDVII obtains merges, through silica gel chromatography, 20-40: the chloroform-methanol mixed solvent eluting of 1 ratio obtains 4-ethyl peoniflorin 0.025-0.045 weight portion; Stream part 31～42 that chromatographic isolation CDV obtains merges, and through purification by silica gel column chromatography, 20-40: the chloroform-methanol eluting of 1 ratio obtains 1S, 2S, 4R-anti-form-1,8-cineole-2-O-β-D-glucoside 0.03-0.05 weight portion;
Portion C E separates with silica gel column chromatography, be chromatographic isolation CEI, petroleum ether-acetone mixed solvent 6-9: 6 eluting, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 28～33 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 19～30 that chromatographic isolation CEI obtains merges, separate with silica gel column chromatography, be chromatographic isolation CEII, 15: 1 → 12: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 18～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, the component 1～6 that chromatographic isolation CEII obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation CEIII, 50: 1 → 20: 1 → 8: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, and every 40-60 parts by volume collection is a, collects altogether to obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 22～28 usefulness Sephadex LH-20 column chromatography purification that chromatographic isolation CEIII obtains, methanol-eluted fractions obtains pyrogallol 0.004-0.008 weight portion; The component 11～12 that chromatographic isolation CEII obtains merges, separate with Sephadex LH-20 column chromatography, be chromatographic isolation CEIV, water → 30% methanol → 50% methanol gradient elution, portioning is collected, every 40-60 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, the component 9～14 that chromatographic isolation CEIV obtains merges, separate i.e. chromatographic isolation CEV, 4～7: 0.5～1.5: the chloroform-methanol of 0.1 ratio-water mixed solvent eluting with polyamide column chromatography, portioning is collected, every 40-60 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, and the inspection of polyamide thin layer chromatography is known the back and merged same stream part, stream part 3～4 that chromatographic isolation CEV obtains merges, with Sephadex LH-20 column chromatography purification, methanol-eluted fractions obtains progallin A 0.005-0.015 weight portion; Stream part 8～15 merging, through the polyamide column chromatography purification, 6-10: the chloroform-methanol mixed solvent eluting of 1 ratio obtains gallic acid 0.015-0.035 weight portion; The component 13～15 that chromatographic isolation CEII obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation CEVI, chloroform-methanol mixed solvent 5-7: 1 eluting, portioning is collected, and every 40-60 parts by volume is collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part; Stream part 14～20 that chromatographic isolation CEVI obtains merges, silica gel column chromatography separates, be chromatographic isolation CEVII, 6-10: the chloroform-methanol mixed solvent eluting of 1 ratio, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 5～9 that chromatographic isolation CEVII obtains merges, separate with reversed-phase silica gel column chromatography, be chromatographic isolation CEVIII, 30% methanol-eluted fractions, portioning is collected, every 10-30 parts by volume is collected a, collect altogether and obtain 15～20 stream parts, the inspection of reverse phase silica gel thin layer chromatography is known the back and is merged same stream part, stream part 4～7 that chromatographic isolation CEVIII obtains merges, separate with silica gel column chromatography, be chromatographic isolation CEVIX, 4～7: 0.5～1.5: the chloroform-methanol of 0.1 ratio-water mixed solvent eluting, portioning is collected, and every 40-60 parts by volume is collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 10～13 that chromatographic isolation CEVIX obtains is separated through reversed-phase silica gel column chromatography, i.e. chromatographic isolation CEVX, 40% methanol-eluted fractions, portioning is collected, and every 10-30 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, the inspection of reverse phase silica gel thin layer chromatography is known the back and is merged same stream part, stream part 14～19 that chromatographic isolation CEVX obtains is through silica gel chromatography, and 6-10: the chloroform-methanol eluting of 1 ratio obtains ethyl lacdtlorin A (25mg); Stream part 14～17 that chromatographic isolation CEVIX obtains merges, separate with silica gel column chromatography, be chromatographic isolation CEVXI, 4～7: 0.5～1.5: the chloroform-methanol of 0.1 ratio-water mixed solvent eluting, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 14～17 merging that chromatographic isolation CEVXI obtains, through Sephadex LH-20 column chromatography purification, methanol-eluted fractions obtains (1S, 2S, 4R)-and anti-form-1,8-cineole-2-O-(6-O-α-L-rhamanopyranosyl)-β '-D-glucoside 0.02-0.04 weight portion;
Portion C F separates through the Toyopearl column chromatography, be chromatographic isolation CFI, the water eluting, portioning is collected, every 10-30 parts by volume is collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 5 usefulness silica gel column chromatographies that chromatographic isolation CFI obtains separate, i.e. chromatographic isolation CFII, 17～23: 1～3: the ethyl acetate, alcohol and water mixed solvent eluting of 1 ratio, portioning is collected, every 40-60 parts by volume collection is a, collects altogether to obtain 20～25 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 13～19 that chromatographic isolation CFII obtains merges, use silica gel chromatography, 17～23: 1～3: the ethyl acetate, alcohol and water eluting of 1 ratio obtains peoniflorin 0.02-0.04 weight portion; Stream part 6～7 that chromatographic isolation CFI obtains merges, separate through silica gel column chromatography, be chromatographic isolation CFIII, 17～23: 1～3: the ethyl acetate, alcohol and water mixed solvent eluting of 1 ratio, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 15～23 that chromatographic isolation CFIII obtains merges, through purification by silica gel column chromatography, 5-15: the chloroform-methanol eluting of 1 ratio obtains Hydroxy peoniflorin 0.035-0.055 weight portion;
The separation of effective ingredient in the Radix Puerariae: get Radix Puerariae 3-18 weight portion, with 30～80% ethanol extractions of 6-10 times of weight portion 1-4 time, each 1-3 hour, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, after being evaporated to the 1500-2500 parts by volume under 70～80 ℃ of conditions of temperature, be diluted with water to the 4000-6000 parts by volume, by AB-8 type macroporous adsorptive resins, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, and collect ethanol elution, decompression and solvent recovery is to doing, residue is put 50～70 ℃ of dryings in the vacuum drying oven, obtains dry extract;
The above-mentioned dry extract 300-500 weight portion of access method preparation carries out silica gel column chromatography to be separated, be chromatographic isolation II, 100: 0 → 20: 1 → 10: 1 → 5: 1 → 2: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 80 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part the 3rd～5,6～7,8～21,22～68,69～80 that chromatographic isolation II is obtained merges respectively, obtains GA, GB, GC, GD, five parts of GE;
Part GA separates through silica gel column chromatography, be chromatographic isolation GAI, the chloroform eluting, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 10 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 2 that chromatographic isolation GAI obtains is separated through Sephadex LH-20 column chromatography, be chromatographic isolation GAII, methanol-eluted fractions, portioning is collected, and every 40-60 parts by volume is collected a, collect altogether and obtain 8～12 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 2～5 that chromatographic isolation GAII obtains merges, and separates through silica gel column chromatography, be chromatographic isolation GAIII, 6-10: the petroleum ether of 1 ratio-acetone mixed solvent eluting, portioning is collected, and every 40-60 parts by volume is collected a, collect altogether and obtain 8～12 stream parts, silica gel thin-layer chromatography is examined and is known back merging same stream part, and the stream part 4 that obtains from chromatographic isolation GAIII obtains Palmic acid 0.005-0.015 weight portion, and stream part 5 obtains arachidic acid 0.01-0.015 weight portion; Stream part 6 is separated out white crystals, uses acetone recrystallization, obtains lupeol 0.004-0.008 weight portion; Stream part 8～9 is separated out white, needle-shaped crystals, uses recrystallizing methanol, obtains cupreol 0.01-0.03 weight portion;
Part GB is separated through silica gel column chromatography, be chromatographic isolation GBI, the chloroform eluting, portioning is collected, every 40-60 parts by volume collection is a, collects altogether to obtain 18～24 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 11～17 that chromatographic isolation GBI obtains merges, separate i.e. chromatographic isolation GBII, 10-30: the chloroform-methanol eluting of 1 ratio with silica gel column chromatography, portioning is collected, every 40-60 parts by volume collection is a, collects altogether to obtain 18～24 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 3 that chromatographic isolation GBII obtains is separated out white precipitate, obtains lignocerane acid glyceride 0.02-0.04 weight portion behind the methanol cyclic washing; Separate out white crystals after stream part 4 placements, use recrystallizing methanol, obtain big legumin 0.025-0.045 weight portion;
Part GC separates through silica gel column chromatography, be chromatographic isolation GCI, with 20: 1 → 10: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 60～70 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 30～42 that chromatographic isolation GCI obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation GCII, 5-15: the chloroform-methanol eluting of 1 ratio, portioning is collected, and every 40-60 parts by volume collection is a, collects altogether to obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 13～20 that chromatographic isolation GCII obtains merges, and therefrom separates out white precipitate, obtains genistin 0.01-0.03 weight portion behind the methanol cyclic washing; Stream part 43～61 that chromatographic isolation GCI obtains merges, separate through silica gel column chromatography, be chromatographic isolation GCIII, 5-15: the chloroform-methanol eluting of 1 ratio, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 13～19 that chromatographic isolation GCIII obtains merges, and uses silica gel chromatography, 5-15: the chloroform-methanol eluting of 1 ratio, obtain 4 ', 8-dimethoxy-7-glucone isoflavone 0.015-0.035 weight portion; Separate out crystallization after stream part that chromatographic isolation GCIII obtains 21～29 is placed, use recrystallizing methanol, obtain 4 '-methoxyl group daiazi 0.035-0.055 weight portion;
Part GD dilutes with suitable quantity of water, by the AB-8 macroporous adsorbent resin, water elution to water lotion near colourless after, be washed till the eluent color with 30～70% ethanol and end when more shallow, the merging ethanol elution, decompression and solvent recovery obtains extractum 20-40 weight portion; Getting above-mentioned extractum separates with silica gel column chromatography, be chromatographic isolation GDI, 100: 0 → 10: 1 → 4: 1 → 2: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 50-150 parts by volume is collected a, collect altogether and obtain 35～40 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 11～19 that chromatographic isolation GDI obtains merges, separate with silica gel column chromatography, be chromatographic isolation GDII, 17-23: 1-3: the ethyl acetate, alcohol and water mixed solvent eluting of 1 ratio, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 2～6 that chromatographic isolation GDII obtains merges, separate with polyamide column chromatography, be chromatographic isolation GDIII, 17-23: 1-3: the ethyl acetate, alcohol and water mixed solvent eluting of 1 ratio, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 15～20 stream parts, the inspection of polyamide thin layer chromatography is known the back and is merged same stream part, and stream part 2～5 that chromatographic isolation GDIII obtains merges, through the polyamide column chromatography purification, 17-23: 1-3: the ethyl acetate, alcohol and water mixed solvent eluting of 1 ratio obtains ononin 0.025 weight portion; 6～14 merge, separate through polyamide column chromatography, be chromatographic isolation GDIV, 25-35: 1-3: the ethyl acetate, alcohol and water mixed solvent eluting of 1 ratio, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 15～20 stream parts, the inspection of polyamide thin layer chromatography is known the back and is merged same stream part, stream part 3～5 that chromatographic isolation GDIV obtains merges, and polyamide column chromatography separates, i.e. chromatographic isolation GDV, 12-18: the chloroform-methanol mixed solvent eluting of 1 ratio, portioning is collected, and every 40-60 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, the inspection of polyamide thin layer chromatography is known the back and is merged same stream part, after placing, stream parts 10 separates out kudzu-vine root new glycoside A 0.015-0.035 weight portion, after placing, stream parts 13～15 separates out 3 '-hydroxyl puerarin 0.005-0.015 weight portion, stream part 17～20 usefulness polyamide column chromatographies separate, 25-35: 1-3: the ethyl acetate, alcohol and water mixed solvent eluting of 1 ratio obtains daiazi 0.01-0.015 weight portion; Stream parts 14～25 merges the back to be placed, and separates out crystallization, uses recrystallizing methanol, obtain 3 '-methoxy puerarin 0.035-0.055 weight portion; Stream part 15～17 that chromatographic isolation GDIII obtains merges, and separates through polyamide column chromatography, and chloroform-methanol mixed solvent 5-10: 1 ratio eluting obtains puerarin 0.045-0.065 weight portion; Stream part 28～36 that chromatographic isolation GDI obtains merges, separate with silica gel column chromatography, be chromatographic isolation GDVI, chloroform-methanol-water mixed solvent 3-8: 0.5-2: 1 ratio eluting, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, separates out crystallization after stream part 13～18 placements that chromatographic isolation GDVI obtains, use recrystallizing methanol, obtain celery glycosyl puerarin 0.04-0.06 weight portion;
The separation of effective ingredient in the Radix Ginseng: get Radix Ginseng 3-18 weight portion, with 30～80% ethanol extractions of 6-10 times of weight portion 1-4 time, each 1-3 hour, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, after being evaporated to the 1500-2500 parts by volume under 70～80 ℃ of conditions of temperature, be diluted with water to the 4000-6000 parts by volume, by AB-8 type macroporous adsorptive resins, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, and collect ethanol elution, decompression and solvent recovery is to doing, residue is put 50～70 ℃ of dryings in the vacuum drying oven, obtains dry extract;
The dry extract 400-800 weight portion of getting method for preparing carries out silica gel column chromatography to be separated, be chromatographic isolation III, 19: 1～0: 1 gradient elution of chloroform-methanol mixed solvent, portioning is collected, every 50-150 parts by volume collection is a, collects altogether to obtain 100 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part the 16th～25,26～40,41～55,56～79,80～100 that chromatographic isolation III is obtained merges respectively, obtains RA, RB, RC, RD, five parts of RE;
Part RB separates with silica gel column chromatography, be chromatographic isolation RBI, chloroform-methanol mixed solvent 15-35: 1 ratio eluting, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 35～40 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 14～22 merging that chromatographic isolation RBI obtains, separate with Sephadex LH-20 column chromatography, be chromatographic isolation RBII, methanol-eluted fractions, portioning is collected, every 15-35 parts by volume is collected a, collect altogether and obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, separates out 20 (S)-ginsenoside Rhs after stream part 13～18 merging that chromatographic isolation RBII obtains 10.035-0.055 weight portion; Separate out 20 (S)-Protopanaxatriol 0.005-0.025 weight portions after stream part 20～23 merging;
Part RC separates with silica gel column chromatography, be chromatographic isolation RCI, chloroform-methanol mixed solvent 5-15: 1 ratio eluting, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 28～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 13～19 that chromatographic isolation RCI obtains merges, and separates with silica gel column chromatography, be chromatographic isolation RCII, chloroform-methanol mixed solvent 5-15: 1 ratio eluting, portioning is collected, and every 40-60 parts by volume is collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 10～16 that chromatographic isolation RCII obtains merges, with Sephadex LH-20 column chromatography purification, methanol-eluted fractions obtains 20 (R)-ginsenoside Rhs 10.02-0.04 weight portion;
Part RD separates with silica gel column chromatography, be chromatographic isolation RDI, chloroform-methanol mixed solvent 5-15: 1 ratio eluting, portioning is collected, every 50ml collects a, collect altogether and obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 16～21 that chromatographic isolation RDI obtains merges, and separates with silica gel column chromatography, be chromatographic isolation RDII, chloroform-methanol mixed solvent 5-15: 1 ratio eluting, portioning is collected, and every 40-60 parts by volume is collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 13～15 that chromatographic isolation RDII obtains merges, and separates through reversed-phase silica gel column chromatography, methanol-eluted fractions, portioning is collected, and every 10-30 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, the reverse phase silica gel thin layer chromatography is examined and is known back merging same stream part, and gravity flow part 13～16 is separated out 20 (S)-ginsenoside Rgs 30.005-0.015 weight portion;
Part RE separates with silica gel column chromatography, be chromatographic isolation REI, chloroform-methanol mixed solvent 5-15: 1 ratio eluting, portioning is collected, and every 40-60 parts by volume is collected a, collect altogether and obtain 83～88 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 27～38 that chromatographic isolation REI obtains merges, through the reversed-phase silica gel column chromatography purification, methanol-eluted fractions obtains the ginsenoside Rg 10.06-0.10 weight portion and 20 (S)-ginsenoside Rg 20.02-0.04 weight portion; Stream part 42～59 merging, separate with silica gel column chromatography, be chromatographic isolation REII, chloroform-methanol mixed solvent 5-15: 1 ratio eluting, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 48～56 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 18～30 that chromatographic isolation REII obtains merges, and separates methanol-eluted fractions through reversed-phase silica gel column chromatography, portioning is collected, every 10-30 parts by volume collection is a, collects altogether to obtain 40～45 stream parts, and the inspection of reverse phase silica gel thin layer chromatography is known the back and merged same stream part, gravity flow part 12～18 obtains ginsenoside Rd 0.04-0.06 weight portion, and gravity flow part obtains ginsenoside Re 0.03-0.05 weight portion for 22～29 parts; Stream part 32～45 that chromatographic isolation REII obtains merges, and through the reversed-phase silica gel column chromatography purification, methanol-eluted fractions obtains ginsenoside Rb 10.04-0.08 weight portion.
The preparation method of pharmaceutical composition active component of the present invention is preferably:
The separation of effective ingredient in the Radix Paeoniae Rubra: get Radix Paeoniae Rubra 6 weight portions, with 30～80% ethanol extractions of 8 times of parts by volume 3 times, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, after being evaporated to 2000 parts by volume under 70～80 ℃ of conditions of temperature, be diluted with water to 5000 parts by volume, by AB-8 type macroporous adsorptive resins, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, and collect ethanol elution, decompression and solvent recovery is to doing, residue is put 50～70 ℃ of dryings in the vacuum drying oven, obtains dry extract;
Above-mentioned dry extract 500 weight portions of access method preparation, carry out silica gel column chromatography and separate (chromatographic isolation I), with 100: 0 → 20: 1 → 10: 1 → 5: 1 → 2: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 115～125 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part the 4th～6,7～16,17～31,32～41,42～74,75～99,100～121 that chromatographic isolation I is obtained merges respectively, obtains CA, CB, CC, CD, CE, CF, seven different pieces of CG;
Portion C A carries out silica gel column chromatography to be separated, and promptly chromatographic isolation CAI uses 2: 1 → 7: 5 gradient elutions of petroleum ether-acetone mixed solvent, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 20～25 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part; Stream part 5～17 that chromatographic isolation CAI obtains merges, silica gel column chromatography separates, and promptly chromatographic isolation CAII uses petroleum ether-acetone mixed solvent 40～60: 1 eluting, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 8～12 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 3～5 that chromatographic isolation CAII obtains merges, separate out the white plates crystallization, use the petroleum ether recrystallization, obtain benzoic acid 0.02 weight portion;
Portion C B carries out silica gel column chromatography to be separated, be chromatographic isolation CBI, with 100: 0 → 50: 1 → 20: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 30～35 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 22～27 that chromatographic isolation CBI obtains merges, separate through silica gel column chromatography, promptly chromatographic isolation CBII uses petroleum ether-acetone mixed solvent (7: 2) eluting, portioning is collected, every 50ml collection is a, collects altogether to obtain 10～15 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 9 that chromatographic isolation CBII obtains is separated through Sephadex LH-20 column chromatography, methanol-eluted fractions obtains 1S, 2S, 4R-is trans-2-hydroxyl-1, and 8-cineole 0.005 weight portion and cupreol 0.02 weight portion;
Portion C D carries out silica gel column chromatography to be separated, be chromatographic isolation CDI, petroleum ether-acetone → methanol gradient elution with the petroleum ether-acetone of 3: 1 ratios → 7.5: 5 ratio, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 75～80 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 41～70 that chromatographic isolation CDI obtains merges, and separates with silica gel column chromatography, be chromatographic isolation CDII, the petroleum ether-ethyl acetate mixed solvent eluting of 2: 1 ratios, portioning is collected, and per 50 parts by volume are collected a, collect altogether and obtain 25～30 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 9～16 that chromatographic isolation CDII obtains merges, and separates through silica gel column chromatography, the petroleum ether of 5: 1 ratios-acetone eluting obtains dihydro apigenin 0.008 weight portion; The methanol-eluted fractions thing that chromatographic isolation CDI obtains separates with silica gel column chromatography, be chromatographic isolation CDIII, 2.5: the petroleum ether of 1 ratio-acetone mixed solvent eluting, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 25～30 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 9～18 that chromatographic isolation CDIII obtains merges, separate with Sephadex LH-20 column chromatography, be chromatographic isolation CDIV, the acetone eluting, portioning is collected, per 30 parts by volume are collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 5～12 that chromatographic isolation CDIV obtains merges, and separates through silica gel column chromatography, i.e. chromatographic isolation CDV, 50: 1 → 20: 1 → 10: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, and per 50 parts by volume collection is a, collects altogether to obtain 45～50 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 8～12 that chromatographic isolation CDV obtains merges, and separates out white crystals, obtains lacdtlorin 0.03 weight portion with acetone recrystallization, stream part 13～30 merging, separate i.e. chromatographic isolation CDVI, 20: 1 → 10: 1 gradient elutions of chloroform-methanol mixed solvent through silica gel column chromatography, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 35～40 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 4～18 that chromatographic isolation CDVI obtains merges, through the preparation of preparation of silica gel chromatographic isolation, the chloroform-methanol mixed solvent of 30: 1 ratios launches, and obtains benzoylpaeoniflorin 0.04 weight portion; Stream part 25～36 merging, separate (chromatographic isolation CDVII) through Sephadex LH-20 column chromatography, the acetone eluting, portioning is collected, and every 20ml collects a, collect altogether and obtain 10～12 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 1～3 that chromatographic isolation CDVII obtains merges, through silica gel chromatography, the chloroform-methanol mixed solvent eluting of 30: 1 ratios obtains 4-ethyl peoniflorin 0.035 weight portion; Stream part 31～42 that chromatographic isolation CDV obtains merges, and through purification by silica gel column chromatography, the chloroform-methanol eluting of 30: 1 ratios obtains 1S, 2S, 4R-anti-form-1,8-cineole-2-O-β-D-glucoside 0.04 weight portion;
Portion C E separates with silica gel column chromatography, be chromatographic isolation CEI, 7.5: 6 eluting of petroleum ether-acetone mixed solvent, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 28～33 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 19～30 that chromatographic isolation CEI obtains merges, separate with silica gel column chromatography, be chromatographic isolation CEII, 15: 1 → 12: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 18～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, the component 1～6 that chromatographic isolation CEII obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation CEIII, 50: 1 → 20: 1 → 8: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, and per 50 parts by volume collection is a, collects altogether to obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 22～28 usefulness Sephadex LH-20 column chromatography purification that chromatographic isolation CEIII obtains, methanol-eluted fractions obtains pyrogallol 0.006 weight portion; The component 11～12 that chromatographic isolation CEII obtains merges, separate with Sephadex LH-20 column chromatography, be chromatographic isolation CEIV, water → 30% methanol → 50% methanol gradient elution, portioning is collected, per 30 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, the component 9～14 that chromatographic isolation CEIV obtains merges, separate i.e. chromatographic isolation CEV, the chloroform-methanol of 5.5: 1.0: 0.1 ratios-water mixed solvent eluting with polyamide column chromatography, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, and the inspection of polyamide thin layer chromatography is known the back and merged same stream part, stream part 3～4 that chromatographic isolation CEV obtains merges, with Sephadex LH-20 column chromatography purification, methanol-eluted fractions obtains progallin A 0.01 weight portion; Stream part 8～15 merging, through the polyamide column chromatography purification, the chloroform-methanol mixed solvent eluting of 8: 1 ratios obtains gallic acid 0.025 weight portion; The component 13～15 that chromatographic isolation CEII obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation CEVI, 6: 1 eluting of chloroform-methanol mixed solvent, portioning is collected, and per 50 parts by volume are collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part; Stream part 14～20 that chromatographic isolation CEVI obtains merges, silica gel column chromatography separates, be chromatographic isolation CEVII, the chloroform-methanol mixed solvent eluting of 8: 1 ratios, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 5～9 that chromatographic isolation CEVII obtains merges, separate with reversed-phase silica gel column chromatography, be chromatographic isolation CEVIII, 30% methanol-eluted fractions, portioning is collected, per 20 parts by volume are collected a, collect altogether and obtain 15～20 stream parts, the inspection of reverse phase silica gel thin layer chromatography is known the back and is merged same stream part, stream part 4～7 that chromatographic isolation CEVIII obtains merges, separate with silica gel column chromatography, be chromatographic isolation CEVIX, 5.5: the chloroform-methanol of 1.0: 0.1 ratios-water mixed solvent eluting, portioning is collected, and per 50 parts by volume are collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 10～13 that chromatographic isolation CEVIX obtains is separated through reversed-phase silica gel column chromatography, i.e. chromatographic isolation CEVX, 40% methanol-eluted fractions, portioning is collected, and per 20 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, the inspection of reverse phase silica gel thin layer chromatography is known the back and is merged same stream part, stream part 14～19 that chromatographic isolation CEVX obtains is through silica gel chromatography, and the chloroform-methanol eluting of 8: 1 ratios obtains ethyl lacdtlorin A (25mg); Stream part 14～17 that chromatographic isolation CEVIX obtains merges, separate with silica gel column chromatography, be chromatographic isolation CEVXI, 5.5: the chloroform-methanol of 1.0: 0.1 ratios-water mixed solvent eluting, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 14～17 merging that chromatographic isolation CEVXI obtains, through Sephadex LH-20 column chromatography purification, methanol-eluted fractions obtains (1S, 2S, 4R)-and anti-form-1,8-cineole-2-O-(6-O-α-L-rhamanopyranosyl)-β-D-glucoside 0.03 weight portion;
Portion C F separates through the Toyopearl column chromatography, be chromatographic isolation CFI, the water eluting, portioning is collected, per 20 parts by volume are collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 5 usefulness silica gel column chromatographies that chromatographic isolation CFI obtains separate, i.e. chromatographic isolation CFII, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 20～25 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 13～19 that chromatographic isolation CFII obtains merges, use silica gel chromatography, the ethyl acetate, alcohol and water eluting of 20: 2: 1 ratios obtains peoniflorin 0.03 weight portion; Stream part 6～7 that chromatographic isolation CFI obtains merges, separate i.e. chromatographic isolation CFIII, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios through silica gel column chromatography, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 20～25 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 15～23 that chromatographic isolation CFIII obtains merges, through purification by silica gel column chromatography, the chloroform-methanol eluting of 10: 1 ratios obtains Hydroxy peoniflorin 0.045 weight portion;
The separation of effective ingredient in the Radix Puerariae: get Radix Puerariae 6 weight portions, with 30～80% ethanol extractions of 8 times of weight portions 3 times, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, after being evaporated to 2000 parts by volume under 70～80 ℃ of conditions of temperature, be diluted with water to 5000 parts by volume, by AB-8 type macroporous adsorptive resins, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, and collect ethanol elution, decompression and solvent recovery is to doing, residue is put 50～70 ℃ of dryings in the vacuum drying oven, obtains dry extract;
Above-mentioned dry extract 400 weight portions of access method preparation carry out silica gel column chromatography to be separated, be chromatographic isolation II, 100: 0 → 20: 1 → 10: 1 → 5: 1 → 2: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 80 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part the 3rd～5,6～7,8～21,22～68,69～80 that chromatographic isolation II is obtained merges respectively, obtains GA, GB, GC, GD, five parts of GE;
Part GA separates through silica gel column chromatography, be chromatographic isolation GAI, the chloroform eluting, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 10 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 2 that chromatographic isolation GAI obtains is separated through Sephadex LH-20 column chromatography, be chromatographic isolation GAII, methanol-eluted fractions, portioning is collected, and per 50 parts by volume are collected a, collect altogether and obtain 8～12 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 2～5 that chromatographic isolation GAII obtains merges, and separates through silica gel column chromatography, be chromatographic isolation GAIII, the petroleum ether of 8: 1 ratios-acetone mixed solvent eluting, portioning is collected, and per 50 parts by volume are collected a, collect altogether and obtain 8～12 stream parts, silica gel thin-layer chromatography is examined and is known back merging same stream part, and the stream part 4 that obtains from chromatographic isolation GAIII obtains Palmic acid 0.01 weight portion, and stream part 5 obtains arachidic acid 0.012 weight portion; Stream part 6 is separated out white crystals, uses acetone recrystallization, obtains lupeol 0.006 weight portion; Stream part 8～9 is separated out white, needle-shaped crystals, uses recrystallizing methanol, obtains cupreol 0.02 weight portion;
Part GB is separated through silica gel column chromatography, be chromatographic isolation GBI, the chloroform eluting, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 18～24 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 11～17 that chromatographic isolation GBI obtains merges, separate i.e. chromatographic isolation GBII, the chloroform-methanol eluting of 20: 1 ratios with silica gel column chromatography, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 18～24 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 3 that chromatographic isolation GBII obtains is separated out white precipitate, obtains lignocerane acid glyceride 0.03 weight portion behind the methanol cyclic washing; Separate out white crystals after stream part 4 placements, use recrystallizing methanol, obtain big legumin 0.035 weight portion;
Part GC separates through silica gel column chromatography, be chromatographic isolation GCI, with 20: 1 → 10: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 60～70 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 30～42 that chromatographic isolation GCI obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation GCII, the chloroform-methanol eluting of 10: 1 ratios, portioning is collected, and per 50 parts by volume collection is a, collects altogether to obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 13～20 that chromatographic isolation GCII obtains merges, and therefrom separates out white precipitate, obtains genistin 0.02 weight portion behind the methanol cyclic washing; Stream part 43～61 that chromatographic isolation GCI obtains merges, and separates through silica gel column chromatography, i.e. chromatographic isolation GCIII, the chloroform-methanol eluting of 10: 1 ratios, portioning is collected, and per 50 parts by volume collection is a, collects altogether to obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 13～19 that chromatographic isolation GCIII obtains merges, and uses silica gel chromatography, the chloroform-methanol eluting of 10: 1 ratios, obtain 4 ', 8-dimethoxy-7-glucone isoflavone 0.025 weight portion; Separate out crystallization after stream part that chromatographic isolation GCIII obtains 21～29 is placed, use recrystallizing methanol, obtain 4 '-methoxyl group daiazi 0.045 weight portion;
Part GD dilutes with suitable quantity of water, by the AB-8 macroporous adsorbent resin, water elution to water lotion near colourless after, be washed till the eluent color with 30～70% ethanol and end when more shallow, the merging ethanol elution, decompression and solvent recovery obtains extractum 30 weight portions; Getting above-mentioned extractum separates with silica gel column chromatography, be chromatographic isolation GDI, 100: 0 → 10: 1 → 4: 1 → 2: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, per 100 parts by volume are collected a, collect altogether and obtain 35～40 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 11～19 that chromatographic isolation GDI obtains merges, separate with silica gel column chromatography, be chromatographic isolation GDII, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 2～6 that chromatographic isolation GDII obtains merges, and separates with polyamide column chromatography, be chromatographic isolation GDIII, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios, portioning is collected, and per 50 parts by volume are collected a, collect altogether and obtain 15～20 stream parts, the inspection of polyamide thin layer chromatography is known the back and is merged same stream part, and stream part 2～5 that chromatographic isolation GDIII obtains merges, through the polyamide column chromatography purification, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios obtains ononin 0.025 weight portion; 6～14 merge, separate through polyamide column chromatography, be chromatographic isolation GDIV, the ethyl acetate, alcohol and water mixed solvent eluting of 30: 2: 1 ratios, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 15～20 stream parts, the inspection of polyamide thin layer chromatography is known the back and is merged same stream part, stream part 3～5 that chromatographic isolation GDIV obtains merges, and polyamide column chromatography separates, i.e. chromatographic isolation GDV, the chloroform-methanol mixed solvent eluting of 15: 1 ratios, portioning is collected, and per 50 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, the inspection of polyamide thin layer chromatography is known the back and is merged same stream part, after placing, stream parts 10 separates out kudzu-vine root new glycoside A 0.025 weight portion, after placing, stream parts 13～15 separates out 3 '-hydroxyl puerarin 0.01 weight portion, stream part 17～20 usefulness polyamide column chromatographies separate, the ethyl acetate, alcohol and water mixed solvent eluting of 30: 2: 1 ratios obtains daiazi 0.012 weight portion; Stream parts 14～25 merges the back to be placed, and separates out crystallization, uses recrystallizing methanol, obtain 3 '-methoxy puerarin 0.045 weight portion; Stream part 15～17 that chromatographic isolation GDIII obtains merges, and separates through polyamide column chromatography, and 7.5: 1 ratio eluting of chloroform-methanol mixed solvent obtain puerarin 0.055 weight portion; Stream part 28～36 that chromatographic isolation GDI obtains merges, separate with silica gel column chromatography, be chromatographic isolation GDVI, 5.5: 1.25: 1 ratio eluting of chloroform-methanol-water mixed solvent, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, separates out crystallization after stream part 13～18 placements that chromatographic isolation GDVI obtains, use recrystallizing methanol, obtain celery glycosyl puerarin 0.05 weight portion;
The separation of effective ingredient in the Radix Ginseng: get Radix Ginseng 6 weight portions, with 30～80% ethanol extractions of 8 times of weight portions 3 times, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, after being evaporated to 2000 parts by volume under 70～80 ℃ of conditions of temperature, be diluted with water to 5000 parts by volume, by AB-8 type macroporous adsorptive resins, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, and collect ethanol elution, decompression and solvent recovery is to doing, residue is put 50～70 ℃ of dryings in the vacuum drying oven, obtains dry extract;
Dry extract 600 weight portions of getting method for preparing carry out silica gel column chromatography to be separated, be chromatographic isolation III, 19: 1～0: 1 gradient elution of chloroform-methanol mixed solvent, portioning is collected, per 100 parts by volume collection is a, collects altogether to obtain 100 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part the 16th～25,26～40,41～55,56～79,80～100 that chromatographic isolation III is obtained merges respectively, obtains RA, RB, RC, RD, five parts of RE;
Part RB separates with silica gel column chromatography, be chromatographic isolation RBI, 25: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 35～40 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 14～22 merging that chromatographic isolation RBI obtains, separate with Sephadex LH-20 column chromatography, be chromatographic isolation RBII, methanol-eluted fractions, portioning is collected, per 25 parts by volume are collected a, collect altogether and obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, separates out 20 (S)-ginsenoside Rhs after stream part 13～18 merging that chromatographic isolation RBII obtains 10.045 weight portion; Separate out 20 (S)-Protopanaxatriols, 0.015 weight portion after stream part 20～23 merging;
Part RC separates with silica gel column chromatography, be chromatographic isolation RCI, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 28～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 13～19 that chromatographic isolation RCI obtains merges, and separates with silica gel column chromatography, be chromatographic isolation RCII, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, and per 50 parts by volume are collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 10～16 that chromatographic isolation RCII obtains merges, with SephadexLH-20 column chromatography purification, methanol-eluted fractions obtains 20 (R)-ginsenoside Rhs 10.03 weight portion;
Part RD separates with silica gel column chromatography, be chromatographic isolation RDI, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, every 50ml collects a, collect altogether and obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 16～21 that chromatographic isolation RDI obtains merges, and separates with silica gel column chromatography, be chromatographic isolation RDII, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, and per 50 parts by volume are collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 13～15 that chromatographic isolation RDII obtains merges, and separates through reversed-phase silica gel column chromatography, methanol-eluted fractions, portioning is collected, and per 20 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, the reverse phase silica gel thin layer chromatography is examined and is known back merging same stream part, and gravity flow part 13～16 is separated out 20 (S)-ginsenoside Rgs 30.01 weight portion;
Part RE separates with silica gel column chromatography, be chromatographic isolation REI, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, and per 50 parts by volume are collected a, collect altogether and obtain 83～88 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 27～38 that chromatographic isolation REI obtains merges, through the reversed-phase silica gel column chromatography purification, methanol-eluted fractions obtains the ginsenoside Rg 10.08 weight portion and 20 (S)-ginsenoside Rg 20.03 weight portion; Stream part 42～59 merging, separate with silica gel column chromatography, be chromatographic isolation REII, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, and per 50 parts by volume collection is a, collects altogether to obtain 48～56 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 18～30 that chromatographic isolation REII obtains merges, and separates methanol-eluted fractions through reversed-phase silica gel column chromatography, portioning is collected, per 20 parts by volume collection is a, collects altogether to obtain 40～45 stream parts, and the inspection of reverse phase silica gel thin layer chromatography is known the back and merged same stream part, gravity flow part 12～18 obtains ginsenoside Rd's 0.05 weight portion, and gravity flow part obtains ginsenoside Re's 0.04 weight portion for 22～29 parts; Stream part 32～45 that chromatographic isolation REII obtains merges, and through the reversed-phase silica gel column chromatography purification, methanol-eluted fractions obtains ginsenoside Rb 10.06 weight portion.
The structure of pharmaceutical composition active component of the present invention is identified as ultraviolet spectra, infrared spectrum, hydrogen nuclear magnetic resonance spectrum, nuclear magnetic resonance of carbon spectrum and mass spectrum etc. all through physics and chemistry constant measuring and method of spectroscopy.Wherein kudzu-vine root new glycoside A is the noval chemical compound of finding first, and the ethyl lacdtlorin is new natural product.4-ethyl peoniflorin, (1S, 2S, 4R)-trans-2-hydroxyl-1,8-cineole, (1S, 2S, 4R)-anti-form-1,8-cineole-2-O-β-D-glucoside, pyrogallol, dihydro apigenin, 4 ', 8-dimethoxy-7-glucone isoflavone, arachidic acid and Palmic acid are for separating the known compound that obtains first from each single medicinal material of the present invention;
Adopt high performance liquid chromatography, confirm to contain in the pharmaceutical composition of the present invention 4-ethyl peoniflorin, ethyl lacdtlorin A, peoniflorin, benzoylpaeoniflorin, lacdtlorin, Hydroxy peoniflorin, (1S, 2S, 4R)-trans-2-hydroxyl-1, the 8-cineole, (1S, 2S, 4R)-anti-form-1,8-cineole-2-O-β-D-glucoside, (1S, 2S, 4R)-anti-form-1,8-cineole-2-O-(6-O-α-L-rhamanopyranosyl)-β-D-glucoside, pyrogallol, benzoic acid, gallic acid, progallin A, the dihydro apigenin, kudzu-vine root new glycoside A, 4 ', 8-dimethoxy-7-glucone isoflavone, puerarin, 3 '-methoxy puerarin, 3 '-the hydroxyl puerarin, genistin, ononin, big legumin, daiazi, 4 '-the methoxyl group daiazi, celery glycosyl puerarin, the lignocerane acid glyceride, Palmic acid, arachidic acid, lupeol, cupreol, the ginsenoside Re, the ginsenoside Rg 1, ginsenoside Rb 1, 20 (S)-ginsenoside Rgs 2, ginsenoside Rd, 20 (S)-ginsenoside Rg 3, 20 (R)-ginsenoside Rhs 1, 20 (S)-ginsenoside Rhs 1, 20 (S)-compositions such as Protopanaxatriol.
Pharmaceutical composition of the present invention and effective site thereof or active component be technology routinely all, add conventional adjuvant, make the dosage form of clinical acceptance, include but not limited to concentrated pill, capsule, drop pill, granule, tablet, soft capsule, slow releasing agent, oral liquid or lyophilized injectable powder.
The ratio of weight portion of the present invention and parts by volume is a grams per milliliter.
Following experiment and embodiment are used to further specify but are not limited to the present invention.
The protective effect of 1 pair of inductive brain function decline mice of D-galactose of experimental example
(1) experiment material
1 animal
The ICR mice, male, body weight 18～20g is provided by Beijing Vital River Experimental Animals Technology Co., Ltd..The quality certification number: SCXK (capital) 2002-0003.
2 medicines and reagent
Be subjected to reagent: pharmaceutical composition effective site of the present invention, be mixed with solution with distilled water before the experiment, 4 ℃ of refrigerators are preserved.
Positive control drug: hydergine tablet, Sandoz Pharma Ltd., state-run Tianjin Hua Jin pharmaceutical factory co-production product.
Reagent: the D-galactose, chemical reagents corporation provides by Beijing.Protein quantification (the bright orchid of Kao Masi), SOD, MDA, GSH test kit, building up bio-engineering research by Nanjing is provided.
3 instruments and equipment
The water maze tester is developed by institute of Materia Medica,Chinese Academy of Medical Sciences electronic machine chamber.HZQ-C air bath agitator, Dongming, Harbin City Medical Instruments factory produces.The 400R refrigerated centrifuge, German Heraeus product.722 type visible spectrophotometers, Shanghai analytical tool company product.
(2) experimental technique and result
1 grouping and administration
Laboratory animal is divided into 6 groups at random, promptly blank group, model control group, pharmaceutical composition effective site of the present invention little, in, heavy dose of group, hydergine tablet group, administration 46 days.Model control group and blank group gavage isometric distilled water.
2 modeling methods
Except that blank group subcutaneous injection every day isometric(al) normal saline, all the other each treated animal equal every day of subcutaneous injection D-galactose 120mg/kg amount to 46 days.
3 assay methods
(1) differentiates learning capacity and measure, adopt the water maze tester, measure the resolution learning capacity of mice in modeling the 43rd, 44,45 days.Began training at the 43rd day, condition is a room temperature (22 ± 2) ℃, and water temperature is (25 ± 1) ℃, depth of water 10cm.Whole experiment divides to be carried out in three days, trained the A point once, B point three times earlier in first day; Measured the B point in second day, training C point three times; Measured the C point on the 3rd day.A, the B point training time is limited in the 2min, and the C point is limited in the 4min.Overtime person is respectively by 2min and 4min.Every each training and testing of mice all writes down time (time of advent) of reaching home and the number of times (errors number) that enters cecum.With the average of training period B, C 2 times of advent and errors number as school grade, with testing period B, C 2 times of advent and errors number as the memory achievement.Parallel carrying out between more than test is all respectively organized measured after after the administration 1 hour.The result sees Table 1 and table 2 respectively.
Table 1 pharmaceutical composition effective site of the present invention is differentiated the influence (x ± s) of learning capacity to the inductive brain function decline mice of D-galactose
Group n ? Dosage (g/kg) B point test result C point test result
The time of advent (second) Errors number The time of advent (second) Errors number
The effective position of model control group blank group hydergine tablet group pharmaceutical composition of the present invention effective position group pharmaceutical composition of the present invention effective position group pharmaceutical composition of the present invention group 14 12 13 ? 15 ? ? 14 ? ? 11 ? ? 0.001 ? 0.05～0.15 ? ? 0.15～0.30 ? ? 0.30～0.50 66.88±38.68 47.14±36.49 * 48.41±33.32 *? 48.11±35.67 *? ? 49.81±37.70 *? ? 54.88±42.77 3.31±3.17 2.08±1.83 * 2.49±2.08 ? 2.20±1.55 *? ? 1.88±1.50 *? ? 2.76±2.50 59.45±57.57 38.92±27.48 * 35.46±31.08 *? 31.58±18.44 **? ? 36.69±36.80 *? ? 36.67±30.96 * 3.29±3.05 2.17±1.68 * 2.13±1.85 * 2.13±1.75 *? ? 1.95±1.56 *? ? 2.21±2.15
Annotate: compare with model control group, *P＜0.05, *P＜0.01
Table 2 pharmaceutical composition effective site of the present invention is differentiated the influence (x ± s) of memory ability to the inductive brain function decline mice of D-galactose
Group n ? Dosage (g/kg) B point test result C point test result
The time of advent (second) Errors number The time of advent (second) Errors number
The effective position of model control group blank group hydergine tablet group pharmaceutical composition of the present invention effective position group pharmaceutical composition of the present invention effective position group pharmaceutical composition of the present invention group 14 12 13 ? 15 ? ? 14 ? ? 11 ? ? ? 0.001 ? 0.05～0.15 ? ? 0.15～0.30 ? ? 0.30～0.50 ? 47.93±32.38 21.83±11.27 * 15.92±12.57 **? 24.87±21.64 *? ? 19.86±15.61 **? ? 26.45±26.36 ? 3.36±3.32 1.00±0.74 * 0.38±0.51 **? 1.27±1.16 *? ? 0.93±1.27 *? ? 1.09±1.22 *? 59.43±56.88 23.75±8.85 * 21.23±12.13 *? 21.8±10.01 *? ? 19.14±8.66 *? ? 23.91±10.25 *? 2.93±2.20 0.75±0.87 ** 0.85±0.99 **? 1.07±1.03 **? ? 0.79±0.80 **? ? 1.18±1.33 *?
Annotate: compare with model control group, *P＜0.05, *P＜0.01
(2) biochemical indicator is measured in modeling the 46th day, behind the administration 1h, breaks end in the ears rear, on 4 ℃ of ice pans, get brain rapidly, remove olfactory bulb, cerebellum and the next brain stem, remaining part adds 9 times of normal saline, and to make 10% brain homogenate standby, to measure SOD vigor, MDA content, GSH content.Operation is all built up the test kit explanation of bio-engineering research institute according to Nanjing and is carried out.The results are shown in Table 3.
Table 3 pharmaceutical composition effective site of the present invention is to the influence of the inductive brain function decline mice of D-galactose biochemical indicator (x ± s)
Group ? n ? Dosage (g/kg) SOD vigor (U/mg) MDA content (μ mol/g) GSH content (mg/g)
The effective position of model control group blank group hydergine tablet group pharmaceutical composition of the present invention effective position group pharmaceutical composition of the present invention effective position group pharmaceutical composition of the present invention group 14 12 13 ? 15 ? ? 14 ? ? 11 ? ? ? 0.001 ? 0.05～0.15 ? ? 0.15～0.30 ? ? 0.30～0.50 ? 115.1±8.76 141.00±39.38 * 159.12±17.17 **? 140.37±11.38 **? ? 143.71±11.25 **? ? 131.18±13.30 **? 3.41±0.77 1.63±0.83 ** 2.41±0.56 **? 1.39±0.49 **? ? 1.91±0.39 **? ? 1.90±0.57 **? 30.58±4.82 48.00±14.15 ** 37.93±4.86 **? 44.59±6.05 **? ? 48.42±15.28 **? ? 58.08±4.69 **?
Annotate: compare with model control group, *P＜0.05, *P＜0.01
(3) conclusion
The subcutaneous injection D-of nape portion galactose can cause the learning and memory of little mouse dysfunction in 46 days, showed as in the water maze laboratory example, and prolong the time of advent, and errors number increases.Pharmaceutical composition effective site of the present invention is little, in, heavy dose all can improve this learning memory disorder in various degree.The subcutaneous injection D-of nape portion galactose can cause mouse brain to organize the biochemical metabolism disorder in 46 days, showed as superoxide dismutase (SOD) vigor and descended, and glutathion (GSH) content reduces, and malonaldehyde (MDA) content raises.Pharmaceutical composition effective site of the present invention is little, in, that heavy dose can obviously be improved this biochemical metabolism is unusual.
2 pairs of experimental examples are intended the protective effect of 8 ℃ of frozen water swimming of blood stasis due to qi deficiency model mice
(1) experimental example material
1 animal
The ICR mice, totally 64, male and female half and half, body weight 18-22 gram is provided by Beijing Vital River Experimental Animals Technology Co., Ltd..
2 medicines
Be subjected to reagent: pharmaceutical composition of the present invention, pharmaceutical composition effective site of the present invention, dissolve wiring solution-forming with 0.5%CMC, 4 ℃ of refrigerators are preserved.
3 equipment
Electronic stopclock, bucket, thermometer etc.
(2) experimental technique and result
1 grouping and administration laboratory animal are divided into five groups at random, 1 group of pharmaceutical composition A promptly of the present invention, 2 groups of pharmaceutical composition A of the present invention, pharmaceutical composition effective site B1 of the present invention group, pharmaceutical composition effective site B2 of the present invention group, model group (0.5%CMC).The continuous gastric infusion of each treated animal 4 days.Administration modeling after 1 hour in the 4th day.
2.5cm place, 2 modeling method mouse tail tip is the rubber mud ball of one 10% body weight, puts into 8 ℃ ± 0.5 ℃ frozen water and swims, and one every barrel, depth of water 15cm, drowned dead until mice.Operation repetitive between five groups, is promptly swum the time-to-live at record this section period of mice from the entry to death.
Relatively carry out the t check between 3 statistical method groups, experimental result is represented with x ± S.
4 experimental examples the results are shown in Table 4.
Table 4 pharmaceutical composition of the present invention is to intending the blood stasis due to qi deficiency model mice influence of swimming time-to-live (x ± S)
Group Dosage (g/kg) n The swimming time-to-live (s)
2 groups of pharmaceutical composition effective site B1 group of the present invention pharmaceutical composition effective site B2 groups of the present invention of 1 group of pharmaceutical composition A of the present invention of normal control group pharmaceutical composition A of the present invention 0.5％CMC 0.8 1.6 0.05～0.25 0.25～0.50 12 12 13 14 13 725.00±105.41 841.83±149.59 * 878.31±234.43 * 796.64±169.24 941.46±224.37 **
Annotate: compare with model group, *P＜0.05, *P＜0.01
(3) conclusion
Pharmaceutical composition of the present invention, pharmaceutical composition effective site of the present invention all have to prolong intends the effect of swimming time-to-live of blood stasis due to qi deficiency model mice, and good with the action effect of pharmaceutical composition effective site of the present invention.
3 pairs of thrombotic influences of rat arteriovenous shut of experimental example
(1) experimental example material
1 animal
The SD rat, male, body weight 288 ± 16g is provided by Beijing Vital River Experimental Animals Technology Co., Ltd., the quality certification number: SCXK (capital) 2002-0003.
2 medicines and reagent
Be subjected to reagent: dissolve wiring solution-forming with 0.5%CMC before the pharmaceutical composition effective site of the present invention, administration, 4 ℃ of refrigerators are preserved.
Positive drug: taponin Folium Ginkgo, pharmaceutical Co. Ltd of Zhejiang Kang Enbei group product dissolves wiring solution-forming with 0.5%CMC before the administration, and 4 ℃ of refrigerators are preserved.
3 instruments
AEG-220 type electronic analytical balance, Japanese Shimadzu company product; DF-206 type air dry oven, west city, Beijing medical apparatus and instruments factory product.
(2) experimental technique and result
1 grouping and administration
Rat is divided into 5 groups at random by body weight, promptly model control group, taponin 24mg/kg group, pharmaceutical composition of the present invention little, in, heavy dose of group.Model group gives isometric(al) 0.5%CMC.The continuous gastric infusion of each treated animal four days, administration in the 4th day experimentized after 1 hour.
2 experimental techniques
The anesthesia of rats by intraperitoneal injection 10% chloral hydrate solution (3.5ml/kg), it is fixing to face upward the position, separate right carotid and left side external jugular vein, three sections polyethylene tubes are linked to each other, one end inserts right common carotid artery, the other end inserts left external jugular vein, middle segment length 10cm, put into one long weighed for 8cm 7 #Surgical thread is filled with normal saline in the pipe, connect right carotid and left external jugular vein, open blood flow is middle Herba Clinopodii after 20 minutes, take out silk thread, put on the template of having weighed and claim weight in wet base, this moment, total weight in wet base deducted former silk thread and template heavily is thrombosed weight in wet base.Place in the baking oven 60 ℃ of bakings 1 hour to constant weight again, cooling is weighed, and this moment, gross dry weight deducted former silk thread and template heavily is thrombosed dry weight, the comparable group differences.
Relatively carry out the t check between 3 statistical method groups, experimental result is represented with x ± S.
4 experimental examples the results are shown in Table 5.
Table 5 pharmaceutical composition of the present invention is to the thrombotic influence of rat experiment (x ± S)
Group n Dosage (g/kg) Wet weight of thrombus (mg) Thrombosis dry weight (mg)
Model control group taponin group pharmaceutical composition effective site of the present invention group pharmaceutical composition effective site of the present invention group pharmaceutical composition effective site of the present invention group 15 15 15 14 14 0.5％CMC 0.024 0.02～0.06 0.06～0.12 0.12～0.24 171.9±41.7 123.6±45.0 ** 135.9±47.0 * 124.7±42.1 ** 131.3±42.9 * 34.2±11.2 24.4±13.6 * 25.5±11.2 * 25.8±7.5 * 22.0±10.4 **
Annotate: compare with model control group, *P＜0.05, *P＜0.01
(3) conclusion
Pharmaceutical composition effective site of the present invention is little, in, heavy dose of group all can alleviate the weight in wet base and the dry weight of rat arteriovenous shut thrombosis, compare (P＜0.05 that has significant difference with model group, P＜0.01), shows that pharmaceutical composition effective site of the present invention has antithrombotic effect.
4 pairs of experimental examples are intended the influence that gas blood stasis virtual model mouse brain is organized energy metabolism
(1) experimental example material
1 animal
The ICR mice, male and female half and half, body weight 20 ± 1g is provided by Beijing Vital River Experimental Animals Technology Co., Ltd., the quality certification number: SCXK (capital) 2002-0003.
2 medicines and reagent
Be subjected to reagent: dissolve wiring solution-forming with 0.5%CMC before the pharmaceutical composition effective site of the present invention, administration, 4 ℃ of refrigerators are preserved.
Positive drug: taponin Folium Ginkgo, pharmaceutical Co. Ltd of Zhejiang Kang Enbei group product.Dissolve wiring solution-forming with 0.5%CMC before the administration, 4 ℃ of refrigerators are preserved.
Reagent: Lactic acid Kit, protein quantification (Coomassie brilliant blue method) test kit, the ATP enzyme reagent kit, the NO test kit, the NOS test kit, building up bio-engineering research by Nanjing provides.
3 instruments
Refrigerated centrifuge, German Heraeus product; The vibration of HZQ-C air is bathed, and Dongming, Harbin City Medical Instruments factory produces; 722 type spectrophotometers, Shanghai the 3rd instrument plant produces.
(2) method and result
1 grouping and administration
Mice is divided into 6 groups at random, i.e. model group, the blank group, positive controls (taponin 40mg/kg), pharmaceutical composition effective site of the present invention is little, in, heavy dose of group.Model group and blank group give equivalent 0.5%CMC.The continuous gastric infusion of each treated animal 4 days, administration modeling after 1 hour in the 4th day.
2 modeling methods
About 2.5cm place, mouse tail tip is the plasticine of one 10% body weight, puts into 8 ℃ ± 0.5 ℃ frozen water respectively and swims, and one every barrel, swims and takes out after 5 minutes.
3 testing indexs
The taking-up mice breaks end immediately and gets brain, rejects brain stem and olfactory bulb, and preparation brain tissue homogenate surveys protein content, LD content, ATP enzyme activity, NO vigor, NOS vigor.
The preparation method of 4 tissue homogenates
The brain that take out the broken end back is rejected brain stem, olfactory bulb, cerebellum adds the homogenate in 4 ℃ of environment of 9 times of normal saline, the tissue homogenate of preparation 10%.
5 protein determination methods
4 ℃ of 10% brain homogenate liquid 1000 are left the heart 10 minutes down, get supernatant, press Coomassie brilliant blue protein reagent box description with 722 type spectrophotometric determination protein contents.
The 6LD assay method
4 ℃ of 10% brain homogenate liquid 1000 are left the heart 5 minutes down, get supernatant, press LD test kit description with 722 type spectrophotometric determination LD content.
The 7ATP enzyme testing method
4 ℃ of 10% brain homogenate liquid 1000 are left the heart 5 minutes down, get supernatant, press ATP enzyme reagent kit description with 722 type spectrophotometric determination ATP enzyme activities.
8NO and NOS assay method
4 ℃ of 10% brain homogenate liquid 1000 are left the heart 5 minutes down, get supernatant, press NO and NOS test kit description with 722 type spectrophotometric determination NO and NOS vigor.
Relatively carry out the t check between 9 statistical method groups, experimental result is represented with x ± S.
10 experimental examples the results are shown in Table 6,7.
The influence of broken end mouth breathing times after table 6 pharmaceutical composition of the present invention is swum to 8 ℃ of frozen water of mice (x ± S)
Group n Dosage (g/kg) The broken end mouth breathing time (S)
The effective position of peaceful group pharmaceutical composition of the present invention effective position group pharmaceutical composition of the present invention effective position group pharmaceutical composition of the present invention group is protected in blank group model group sky 17 16 15 16 16 16 0.5％CMC 0.5％CMC 0.04 0.05～0.15 0.15～0.30 0.30～0.50 15.41±1.94 ** 29.69±4.53 34.53±3.52 ** 33.81±5.32 * 34.38±5.18 * 35.75±4.92 **
Annotate: compare with model control group, *P＜0.05, *P＜0.01
The influence of cerebral tissue LD, ATP enzyme activity after table 7 pharmaceutical composition effective site of the present invention is swum to 8 ℃ of frozen water of mice (x ± S)
Group ? n ? Dosage (g/kg) LD content (μ mol/L) Na +-K +-ATP enzyme activity (U/ml) Ca 2+-Mg 2+-ATP enzyme activity (U/ml)
The effective position of peaceful group pharmaceutical composition of the present invention effective position group pharmaceutical composition of the present invention effective position group pharmaceutical composition of the present invention group is protected in blank group model group sky 17 16 15 ? 16 ? ? 16 ? ? 16 ? 0.5％CMC 0.5％CMC 0.04 ? 0.05～0.15 ? ? 0.15～0.30 ? ? 0.30～0.50 ? 0.253±0.049 ** 0.430±0.080 0.344±0.114 *? 0.342±0.070 **? ? 0.311±0.076 **? ? 0.298±0.067 **? 1.073±0.298 ** 0.613±0.194 0.971±0.295 **? 0.819±0.095 **? ? 1.184±0.211 **? ? 1.318±0.296 **? 0.703±0.202 * 0.573±0.119 0.800±0.212 **? 0.964±0.278 **? ? 1.504±0.359 **? ? 1.859±0.600 **?
Annotate: compare with model control group, *P＜0.05, *P＜0.01
(3) conclusion
Pharmaceutical composition effective site of the present invention is little, in, heavy dose of group, can obviously prolong the mice broken end mouth breathing time, compare have significant difference (P＜0.01, P＜0.05=with model group.
Mice is after 8 ℃ of frozen water swimming, and the cerebral tissue energy metabolism occurs unusual, and showing as the cerebral tissue lactic acid content increases Na +-K +-ATP enzyme and Ca 2+-Mg 2+-ATP enzyme activity descends, and compares with the blank group to have significant difference.Pharmaceutical composition effective site of the present invention is little, in, heavy dose of group all can obviously suppress frozen water swimming and cause mouse brain and organize lactic acid content to increase Na +-K +-ATP enzyme and Ca 2+-Mg 2+-ATP enzyme activity descends, and compares with model group to have significant difference (P＜0.05, P＜0.01).Illustrate that pharmaceutical composition of the present invention can produce cerebral protection to the blood stasis due to qi deficiency model mice.
The influence of 5 pairs of bilateral ligation learning and memory in rats of experimental example ability
(1) materials and methods
1 animal
The SD rat, male, body weight 240-260g is provided by Beijing dimension tonneau China logical thing center of experiment, and the quality certification number is: SCXK (capital): 2002-0003.
2 medicines and reagent
Be subjected to reagent: dissolve wiring solution-forming with 0.5%CMC before the pharmaceutical composition effective site of the present invention, administration, 4 ℃ of refrigerators are preserved.
Positive drug: hydergine, by the co-production of the state-run Hua Jin in Sandoz Pharma Ltd. pharmaceutical factory.Dissolve wiring solution-forming with 0.5%CMC before the administration, 4 ℃ of refrigerators are preserved.
Reagent: the NO test kit, the NOS test kit, the Ache test kit, building up bio-engineering research by Nanjing is provided.
3 instruments
The 400R refrigerated centrifuge, German product; The air bath agitator, Dongming, Harbin City Medical Instruments factory produces; 722 type spectrophotometers, Shanghai the 3rd analytical tool factory produces; Rat water maze tester is provided by institute of Materia Medica,Chinese Academy of Medical Sciences electronic machine chamber.
(2) method and result
1 grouping and administration:
Rat is divided into 6 groups at random, i.e. sham operated rats, model control group, positive drug hydergine group, pharmaceutical composition effective site of the present invention is little, in, heavy dose of group.Sham operated rats and model control group give the 0.5%CMC of equivalent.Each treated animal successive administration 44 days, administration modeling after 1 hour in the 4th day.
2 modeling methods:
Experimental rat is with 10% chloral hydrate 3.5ml/kg intraperitoneal injection of anesthesia, and is fixing, sterilization, aseptic operation.Separate bilateral common carotid arteries, carry out permanent ligation with trumpeter's art silk thread, suture muscles skin, lumbar injection gentamycin sulfate, 0.4ml/ are only.Sham operated rats except that not ligation bilateral common carotid arteries, surplus same model group.Postoperative gastric infusion every day continuous 40 days, carried out the water maze behavior determination in last five days in administration.
The behavior of 3Morris water maze detects:
Water maze is a rustless steel round pool, diameter 120cm, high 50cm, by four place of entry of pool wall the pond is divided into the southeast, southwest, four quadrants in northwest and northeast, place a diameter 9.5cm, the circular transparent platform of high 26.5cm apart from pool wall 22cm place in the Southwest Quadrant center.Laboratory temperature remains on about 26 ℃, in the labyrinth, add two bags of milk powder (400g/ bag), wash open with hot water, add water to again and exceed platform 1cm, water temperature remains on 23 ℃ ± 0.5 ℃, is hung with video camera directly over the labyrinth, wide-angle zoom lens, and the connection monitor, be inserted with record such as the computer display system of image pick-up card and data acquisition of Morris water maze and analysis software.With the hair dye blacking of experimental rat head, can train after drying, training period keeps the environment peace and quiet, and object of reference remains unchanged outside the labyrinth.Training comprises four days orientation navigation experiment example and one day space search experimental example:
(1) orientation navigation experiment example:
Be used to measure the acquisition capability of rat to the water maze learning and memory.Experiment lasts four days, and animal subject every day train successively respectively from northeast by two the quadrant place of entry in northwest, rat is put into water towards pool wall,, write down its actual escape latency if rat is found platform in 60s, if do not find platform in 60s, then escape latency is designated as 60s.
(2) space search experimental example:
After being used to measure rat association searching platform, to the hold facility of platform space position memory.After the orientation navigation experiment example finishes, removed platform in the 5th day, put into water by the rat of naming a person for a particular job in the offside quadrant (being Northeast Quadrant) towards pool wall, survey its time of spanning platform first in 60s, pass through the number of times of original platform relevant position and account for the percentage ratio of distance in the swimming distance of platform place quadrant.
4 biochemical indicators are measured:
After rat is finished the water maze behavior determination, get blood and carry out biochemical measurement.
(1) NO, the NOS assay method:
The rat whole blood is left the heart 8 minutes for 4 ℃ 3000, get respectively serum according to the test kit description with 722 type spectrophotometric determination content.
(2) Ache assay method:
The rat whole blood is left the heart 8 minutes for 4 ℃ 3000, get serum according to the test kit description with 722 type spectrophotometric determination content.
5 statistical method: relatively carry out the t check between group, experimental result is represented with x ± S.
6 experimental example results: see Table 8,9,10.
(1) to the influence of bilateral ligation learning and memory in rats ability
The orientation navigation experiment example
Table 8 pharmaceutical composition effective site of the present invention was to bilateral ligation rat four days
The influence of escape latency in the orientation navigation experiment example (x ± S)
Group n ? Dosage g/kg Escape latency (s)
First day Second day The 3rd day The 4th day
The effective position of sham-operation group model group hydergene group pharmaceutical composition of the present invention effective position group pharmaceutical composition of the present invention effective position group pharmaceutical composition of the present invention group 14 16 13 ? 18 ? 16 15 ? 0.5％CMC 0.5％CMC 0.0006 ? 0.02～0.06 ? 0.06～0.12 0.12～0.24 ? 47.5±20.72 53.6±15.89 58.28±6.62 ? 57.70±8.77 ? 54.6±15.89 54.3±15.29 ? 44.48±22.11 46.63±17.41 47.69±18.76 ? 52.01±15.43 ? 43.96±19.95 45.40±20.60 ? 35.95±22.23 36.50±24.12 42.34±20.07 ? 40.03±22.67 ? 35.75±24.78 41.74±21.66 ? 31.79±20.84 * 42.70±19.01 30.54±23.90 *? 34.23±23.09 ? 39.72±22.47 30.99±23.44 *?
Annotate: compare with model group, *P＜0.05, *P＜0.01
The space search experimental example
Table 9 pharmaceutical composition effective site of the present invention in the bilateral ligation rat space search experimental example for the first time the spanning platform time,
Spanning platform number of times, platform place quadrant swimming distance always accounts for influence apart from percentage ratio (x ± S)
Group ? n ? Dosage g/kg The spanning platform time (S) first time The spanning platform number of times The swimming of original platform quadrant is apart from percentage ratio (%)
The effective position of sham-operation group model group hydergene group pharmaceutical composition of the present invention effective position group pharmaceutical composition of the present invention effective position group pharmaceutical composition of the present invention group 14 16 13 18 ? 16 ? 15 0.5％CMC 0.5％CMC 0.0006 0.02～0.06 ? 0.06～0.12 ? 0.12～0.24 42.15±20.40 * 51.79±15.85 38.69±23.69 * 46.67±20.38 ? 43.85±20.49 ? 41.93±22.43 * 0.93±1.08 ** 0.31±0.54 0.50±0.58 0.47±0.65 ? 0.81±1.00 *? 0.70±0.88 * 28.55±10.38 * 23.16±7.76 31.13±14.56 ** 28.49±7.92 **? 27.00±9.05 ? 28.50±9.07 *
Annotate: compare with model group, *P＜0.05, *P＜0.01 (2) is to the influence of ligation bilateral common carotid arteries rat blood serum NO, NOS, Ache
Table 10 pharmaceutical composition effective site of the present invention is to the influence of ligation bilateral common carotid arteries rat blood serum NO, NOS, Ache (x ± S)
Group ? n Dosage (g/kg) NO content (μ mol/L) NOS vigor (U/ml) Ache vigor (U/ml)
The effective position of sham-operation group model group hydergene group pharmaceutical composition of the present invention effective position group pharmaceutical composition of the present invention effective position group pharmaceutical composition of the present invention group 13 13 12 13 ? 13 ? 13 0.5％CMC 0.5％CMC 0.0006 0.02～0.06 ? 0.06～0.12 ? 0.12～0.24 19.01±5.60 * 25.17±8.48 18.63±4.45 * 24.27±6.91 ? 18.89±4.93 *? 18.50±4.67 * 32.12±2.24 * 29.16±4.33 34.05±2.57 36.43±7.47 **? 37.27±2.84 **? 41.12±4.00 ** 8.85±2.40 ** 26.40±3.48 12.88±2.09 ** 25.96±5.21 ? 18.47±5.94 **? 13.97±4.52 **
Annotate: compare with model control group, *P＜0.05, *P＜0.01
(3) conclusion
Adopt 40 days method of the permanent ligation of bilateral common carotid arteries can make rat learning memory disorder occur.Behavioristics's experimental result shows that the continuous after surgery gastric infusion of pharmaceutical composition effective site of the present invention 44 days can make the rat model learning memory disorder make moderate progress.The biochemical indicator measurement result shows that pharmaceutical composition effective site of the present invention can influence NO in the permanent ligation rat blood serum of bilateral common carotid arteries, NOS, AchE vigor.
The influence of blood stasis due to qi deficiency model mice clotting time is intended in 6 pairs of 8 ℃ of frozen water swimming of experimental example
(1) materials and methods
1 animal
The ICR mice, male, body weight 17-19g is provided by Beijing Vital River Experimental Animals Technology Co., Ltd., the quality certification number: SCXK (capital) 2002-0003.
2 medicines
Be subjected to reagent: dissolve wiring solution-forming with 0.5%CMC before the pharmaceutical composition effective site of the present invention, administration, 4 ℃ of refrigerators are preserved.
Positive control drug: taponin Folium Ginkgo, brown tablet.Zhejiang Kang Enbei pharmaceutical Co. Ltd product.Use the dissolved in distilled water wiring solution-forming, 4 ℃ of freezer storages before the experiment.
(2) experimental technique and result
1 grouping and administration
Laboratory animal is divided into five groups at random, i.e. model control group, the taponin group, pharmaceutical composition effective site of the present invention is little, in, heavy dose of group.The continuous gastric infusion of each treated animal four days, administration modeling after 1 hour in the 4th day, wherein model control group gives isometric distilled water.
2 modeling methods
Mice is put into 8 ℃ ± 0.5 ℃ frozen water swimming 5 minutes, takes out.Room temperature keeps 25 ℃.
The mensuration of 3 clotting times
After mice is taken out in frozen water, get blood with capillary tube in the mice ophthalmic corner of the eyes immediately, record treating the preponderant disease instead of the secondary disease blood begins a period of time to blood coagulation as clotting time.
4 statistical method: relatively carry out the t check between group, experimental result is represented with x ± S.
5 experimental example results: see Table 11.
Table 11 pharmaceutical composition effective site of the present invention to 8 ℃ of frozen water swimming intend the influence (x ± S) of blood stasis due to qi deficiency model mice clotting times
Group Dosage (g/kg) n Clotting time (s)
Model control group taponin group pharmaceutical composition effective site of the present invention group pharmaceutical composition effective site of the present invention group pharmaceutical composition effective site of the present invention group ? 0.04 0.05～0.15 0.15～0.30 0.30～0.50 14 14 14 14 13 150.64±48.33 208.64±38.64 ** 146.5±25.68 153.86±27.34 237.00±38.61 **
Annotate: compare with model control group, *P＜0.01
(3) conclusion
Pharmaceutical composition effective site 0.30～0.50g/kg of the present invention can obviously prolong plan blood stasis due to qi deficiency model mice clotting time, compares with model group to have significant difference, (P＜0.05, P＜0.01).
The influence of 7 pairs of middle cerebral artery thrombosiss of experimental example (MCAT) rat model nervous symptoms, cerebral infarction scope and edema degree
Experimental results show that: MCAT rat nervous symptoms scoring score value increases, and the cerebral infarction scope increases, and degree of cerebral edema improves.Pharmaceutical composition effective site of the present invention is little, in, heavy dose of group all can improve MCAT rat nervous symptoms, reduces its cerebral infarction scope; Obviously reduce MCAT rat degree of cerebral edema.Illustrate that it has the effect of antagonism cerebral ischemia.
Following embodiment all can realize the effect of above-mentioned experimental example.
The specific embodiment
Embodiment 1:
Radix Ginseng 10Kg, Radix Puerariae 10Kg, Radix Paeoniae Rubra 10Kg
The above-mentioned raw materials medicine adds conventional adjuvant, according to common process, makes capsule.
Embodiment 2:
Radix Ginseng 3Kg, Radix Puerariae 18Kg, Radix Paeoniae Rubra 9Kg
The above-mentioned raw materials medicine adds conventional adjuvant, according to common process, makes injection.
Embodiment 3:
Radix Ginseng 18Kg, Radix Puerariae 3Kg, Radix Paeoniae Rubra 12Kg
The above-mentioned raw materials medicine adds conventional adjuvant, according to common process, makes oral liquid.
Embodiment 4:
Radix Ginseng 6Kg, Radix Puerariae 15Kg, Radix Paeoniae Rubra 12Kg
The above-mentioned raw materials medicine merges decocting in water, and precipitate with ethanol filters, and concentrates, and the gained mixture is added conventional adjuvant, according to common process, makes oral liquid.
Embodiment 5:
Radix Ginseng 15Kg, Radix Puerariae 6Kg, Radix Paeoniae Rubra 12Kg
The above-mentioned raw materials medicine is decocting in water respectively, and precipitate with ethanol is crossed macroporous resin column absorption, filters respectively, concentrate respectively extract, with extract obtained mixing, add conventional adjuvant, according to common process, make injection.
Embodiment 6:
Radix Ginseng 6Kg, Radix Puerariae 12Kg, Radix Paeoniae Rubra 15Kg
The above-mentioned raw materials medicine is decocting in water respectively, and precipitate with ethanol filters, concentrate respectively extract, with extract obtained mixing, add conventional adjuvant, according to common process, make capsule.
Embodiment 7:
Radix Ginseng 10kg, Radix Puerariae 10kg, Radix Paeoniae Rubra 10kg
Weighting raw materials according to the above ratio, mix homogeneously, 30～80% ethanol extractions 3 times that add 8L, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, be evaporated to 2 times of volumes of medical material amount under 70～80 ℃ of conditions of temperature, relative density is about 1.04～1.05 concentrated solution under 60～65 ℃ of conditions, adsorbs by AB-8 type macroporous adsorbent resin, and liquid to be extracted is all by behind the resin column, water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting, collect ethanol elution with 30～80% ethanol, decompression and solvent recovery promptly gets pharmaceutical composition effective site of the present invention to doing;
This effective site adds conventional adjuvant, according to common process, make concentrated pill.
Embodiment 8:
Radix Ginseng 3kg, Radix Puerariae 18kg, Radix Paeoniae Rubra 9kg
Take by weighing Radix Ginseng, 30～80% ethanol extractions 3 times that add 7L, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, is evaporated to 2 times of volumes of medical material amount under 70～80 ℃ of conditions of temperature, and relative density is about 1.04～1.05 concentrated solution under 60～65 ℃ of conditions, adsorb by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery gets the Radix Ginseng extract part to doing;
Take by weighing Radix Puerariae, 30～80% ethanol extractions 3 times that add 9L, each 2 hours, merge each time extracting solution, concentrating under reduced pressure (please describe spissated condition in detail, and be concentrated into? the qualification of ml or g), concentrated solution adsorbs by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery gets the Radix Puerariae extract part to doing;
Take by weighing Radix Paeoniae Rubra, 30～80% ethanol extractions 3 times that add 7L, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, is evaporated to 2 times of volumes of medical material amount under 70～80 ℃ of conditions of temperature, and relative density is about 1.04～1.05 concentrated solution under 60～65 ℃ of conditions, adsorb by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery gets the Radix Paeoniae Rubra extract part to doing;
Get above-mentioned Radix Ginseng extract part, Radix Puerariae extract part and Radix Paeoniae Rubra extract part, by 1: 1: 1,1: 20: 1,1: 1: 20,1: 10: 10 or 3: 4: 3 mixed were even, promptly get pharmaceutical composition effective site of the present invention;
This effective site adds conventional adjuvant, according to common process, make lyophilized injectable powder.
Embodiment 9:
Radix Ginseng 18Kg, Radix Puerariae 3Kg, Radix Paeoniae Rubra 12Kg
Weighting raw materials according to the above ratio, mix homogeneously, 30～80% ethanol extractions 3 times that add 8L, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, be evaporated to 2 times of volumes of medical material amount under 70～80 ℃ of conditions of temperature, relative density is about 1.04～1.05 concentrated solution under 60～65 ℃ of conditions, adsorbs by AB-8 type macroporous adsorbent resin, and liquid to be extracted is all by behind the resin column, water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting, collect ethanol elution with 30～80% ethanol, decompression and solvent recovery promptly gets pharmaceutical composition effective site of the present invention to doing;
This effective site adds conventional adjuvant, according to common process, make oral liquid.
Embodiment 10:
Radix Ginseng 18Kg, Radix Puerariae 3Kg, Radix Paeoniae Rubra 12Kg
Take by weighing Radix Ginseng, 30～80% ethanol extractions 3 times that add 8L, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, is evaporated to 2 times of volumes of medical material amount under 70～80 ℃ of conditions of temperature, and relative density is about 1.04～1.05 concentrated solution under 60～65 ℃ of conditions, concentrated solution adsorbs by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery gets the Radix Ginseng extract part to doing;
Take by weighing Radix Puerariae, 30～80% ethanol extractions 3 times that add 8L, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, is evaporated to 2 times of volumes of medical material amount under 70～80 ℃ of conditions of temperature, and relative density is about 1.04～1.05 concentrated solution under 60～65 ℃ of conditions, concentrated solution adsorbs by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery gets the Radix Puerariae extract part to doing;
Take by weighing Radix Paeoniae Rubra, 30～80% ethanol extractions 3 times that add 8L, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, is evaporated to 2 times of volumes of medical material amount under 70～80 ℃ of conditions of temperature, and relative density is about 1.04～1.05 concentrated solution under 60～65 ℃ of conditions, concentrated solution adsorbs by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery gets the Radix Paeoniae Rubra extract part to doing;
Get above-mentioned Radix Ginseng extract part, Radix Puerariae extract part and Radix Paeoniae Rubra extract part, by 1: 1: 1,1: 20: 1,1: 1: 20,1: 10: 10 or 3: 4: 3 mixed were even, promptly get pharmaceutical composition effective site of the present invention;
This effective site adds conventional adjuvant, according to common process, make injection.
Embodiment 11:
Radix Ginseng 6g, Radix Puerariae 6g, Radix Paeoniae Rubra 6g
The separation of effective ingredient in the Radix Paeoniae Rubra: get Radix Paeoniae Rubra 6g, with 30～80% ethanol extractions of 8 times of parts by volume 3 times, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, be evaporated to 2000ml under 70～80 ℃ of conditions of temperature after, be diluted with water to 5000ml, by AB-8 type macroporous adsorptive resins, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, and collect ethanol elution, decompression and solvent recovery is to doing, residue is put 50～70 ℃ of dryings in the vacuum drying oven, obtains dry extract;
Get the dry extract 500g of method for preparing, carry out silica gel column chromatography and separate (chromatographic isolation I), with 100: 0 → 20: 1 → 10: 1 → 5: 1 → 2: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 50ml collects a, collect altogether and obtain 115～125 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part the 4th～6,7～16,17～31,32～41,42～74,75～99,100～121 that chromatographic isolation I is obtained merges respectively, obtains CA, CB, CC, CD, CE, CF, seven different pieces of CG;
Portion C A carries out silica gel column chromatography to be separated, and promptly chromatographic isolation CAI uses 2: 1 → 7: 5 gradient elutions of petroleum ether-acetone mixed solvent, and portioning is collected, and every 50ml collection is a, and collection obtains 20～25 stream parts altogether, merges same stream part after the silica gel thin-layer chromatography inspection is known; Stream part 5～17 that chromatographic isolation CAI obtains merges, silica gel column chromatography separates, and promptly chromatographic isolation CAII uses petroleum ether-acetone mixed solvent 40～60: 1 eluting, portioning is collected, every 50ml collection is a, collects altogether to obtain 8～12 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 3～5 that chromatographic isolation CAII obtains merges, separate out the white plates crystallization, use the petroleum ether recrystallization, obtain benzoic acid 0.02g;
Portion C B carries out silica gel column chromatography to be separated, be chromatographic isolation CBI, with 100: 0 → 50: 1 → 20: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 50ml collection is a, collects altogether to obtain 30～35 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 22～27 that chromatographic isolation CBI obtains merges, separate through silica gel column chromatography, promptly chromatographic isolation CBII uses petroleum ether-acetone mixed solvent (7: 2) eluting, portioning is collected, every 50ml collection is a, collects altogether to obtain 10～15 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 9 that chromatographic isolation CBII obtains is separated through Sephadex LH-20 column chromatography, methanol-eluted fractions obtains 1S, 2S, 4R-is trans-2-hydroxyl-1, and 8-cineole 0.005g and cupreol 0.02g;
Portion C D carries out silica gel column chromatography to be separated, be chromatographic isolation CDI, petroleum ether-acetone → methanol gradient elution with the petroleum ether-acetone of 3: 1 ratios → 7.5: 5 ratio, portioning is collected, every 50ml collects a, collect altogether and obtain 75～80 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 41～70 that chromatographic isolation CDI obtains merges, and separates with silica gel column chromatography, be chromatographic isolation CDII, the petroleum ether-ethyl acetate mixed solvent eluting of 2: 1 ratios, portioning is collected, and every 50ml collects a, collect altogether and obtain 25～30 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 9～16 that chromatographic isolation CDII obtains merges, and separates through silica gel column chromatography, the petroleum ether of 5: 1 ratios-acetone eluting obtains dihydro apigenin 0.008g; The methanol-eluted fractions thing that chromatographic isolation CDI obtains separates with silica gel column chromatography, be chromatographic isolation CDIII, 2.5: the petroleum ether of 1 ratio-acetone mixed solvent eluting, portioning is collected, every 50ml collects a, collect altogether and obtain 25～30 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 9～18 that chromatographic isolation CDIII obtains merges, separate with Sephadex LH-20 column chromatography, be chromatographic isolation CDIV, the acetone eluting, portioning is collected, every 30ml collects a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 5～12 that chromatographic isolation CDIV obtains merges, and separates through silica gel column chromatography, i.e. chromatographic isolation CDV, 50: 1 → 20: 1 → 10: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, and every 50ml collection is a, collects altogether to obtain 45～50 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 8～12 that chromatographic isolation CDV obtains merges, and separates out white crystals, obtains lacdtlorin 0.03g with acetone recrystallization, stream part 13～30 merging, separate i.e. chromatographic isolation CDVI, 20: 1 → 10: 1 gradient elutions of chloroform-methanol mixed solvent through silica gel column chromatography, portioning is collected, every 50ml collection is a, collects altogether to obtain 35～40 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 4～18 that chromatographic isolation CDVI obtains merges, through the preparation of preparation of silica gel chromatographic isolation, the chloroform-methanol mixed solvent of 30: 1 ratios launches, and obtains benzoylpaeoniflorin 0.04g; Stream part 25～36 merging, separate (chromatographic isolation CDVII) through Sephadex LH-20 column chromatography, the acetone eluting, portioning is collected, and every 20ml collects a, collect altogether and obtain 10～12 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 1～3 that chromatographic isolation CDVII obtains merges, through silica gel chromatography, the chloroform-methanol mixed solvent eluting of 30: 1 ratios obtains 4-ethyl peoniflorin 0.035g; Stream part 31～42 that chromatographic isolation CDV obtains merges, and through purification by silica gel column chromatography, the chloroform-methanol eluting of 30: 1 ratios obtains 1S, 2S, 4R-anti-form-1,8-cineole-2-O-β-D-glucoside 0.04g;
Portion C E separates with silica gel column chromatography, be chromatographic isolation CEI, 7.5: 6 eluting of petroleum ether-acetone mixed solvent, portioning is collected, and every 50ml collection is a, collects altogether to obtain 28～33 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 19～30 that chromatographic isolation CEI obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation CEII, 15: 1 → 12: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, and every 50ml collection is a, collects altogether to obtain 18～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, the component 1～6 that chromatographic isolation CEII obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation CEIII, 50: 1 → 20: 1 → 8: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, and every 50ml collection is a, collects altogether to obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 22～28 usefulness Sephadex LH-20 column chromatography purification that chromatographic isolation CEIII obtains, methanol-eluted fractions obtains pyrogallol 0.006g; The component 11～12 that chromatographic isolation CEII obtains merges, separate with Sephadex LH-20 column chromatography, be chromatographic isolation CEIV, water → 30% methanol → 50% methanol gradient elution, portioning is collected, every 30ml collection is a, collects altogether to obtain 15～20 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, the component 9～14 that chromatographic isolation CEIV obtains merges, separate i.e. chromatographic isolation CEV, the chloroform-methanol of 5.5: 1.0: 0.1 ratios-water mixed solvent eluting with polyamide column chromatography, portioning is collected, every 50ml collection is a, collects altogether to obtain 15～20 stream parts, and the inspection of polyamide thin layer chromatography is known the back and merged same stream part, stream part 3～4 that chromatographic isolation CEV obtains merges, with Sephadex LH-20 column chromatography purification, methanol-eluted fractions obtains progallin A 0.01g; Stream part 8～15 merging, through the polyamide column chromatography purification, the chloroform-methanol mixed solvent eluting of 8: 1 ratios obtains gallic acid 0.025g; The component 13～15 that chromatographic isolation CEII obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation CEVI, 6: 1 eluting of chloroform-methanol mixed solvent, portioning is collected, and every 50ml collects a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part; Stream part 14～20 that chromatographic isolation CEVI obtains merges, silica gel column chromatography separates, be chromatographic isolation CEVII, the chloroform-methanol mixed solvent eluting of 8: 1 ratios, portioning is collected, every 50ml collects a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 5～9 that chromatographic isolation CEVII obtains merges, separate with reversed-phase silica gel column chromatography, be chromatographic isolation CEVIII, 30% methanol-eluted fractions, portioning is collected, every 20ml collects a, collect altogether and obtain 15～20 stream parts, the inspection of reverse phase silica gel thin layer chromatography is known the back and is merged same stream part, and stream part 4～7 that chromatographic isolation CEVIII obtains merges, and separates with silica gel column chromatography, be chromatographic isolation CEVIX, 5.5: the chloroform-methanol of 1.0: 0.1 ratios-water mixed solvent eluting, portioning is collected, and every 50ml collects a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 10～13 that chromatographic isolation CEVIX obtains is separated through reversed-phase silica gel column chromatography, i.e. chromatographic isolation CEVX, 40% methanol-eluted fractions, portioning is collected, and every 20ml collection is a, collects altogether to obtain 15～20 stream parts, the inspection of reverse phase silica gel thin layer chromatography is known the back and is merged same stream part, stream part 14～19 that chromatographic isolation CEVX obtains is through silica gel chromatography, and the chloroform-methanol eluting of 8: 1 ratios obtains ethyl lacdtlorin A (25mg); Stream part 14～17 that chromatographic isolation CEVIX obtains merges, separate with silica gel column chromatography, be chromatographic isolation CEVXI, 5.5: the chloroform-methanol of 1.0: 0.1 ratios-water mixed solvent eluting, portioning is collected, every 50ml collects a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 14～17 merging that chromatographic isolation CEVXI obtains, through Sephadex LH-20 column chromatography purification, methanol-eluted fractions obtains (1S, 2S, 4R)-and anti-form-1,8-cineole-2-O-(6-O-α-L-rhamanopyranosyl)-β-D-glucoside 0.03g;
Portion C F separates through the Toyopearl column chromatography, be chromatographic isolation CFI, the water eluting, portioning is collected, every 20ml collects a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 5 usefulness silica gel column chromatographies that chromatographic isolation CFI obtains separate, i.e. chromatographic isolation CFII, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios, portioning is collected, every 50ml collection is a, collects altogether to obtain 20～25 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 13～19 that chromatographic isolation CFII obtains merges, use silica gel chromatography, the ethyl acetate, alcohol and water eluting of 20: 2: 1 ratios obtains peoniflorin 0.03g; Stream part 6～7 that chromatographic isolation CFI obtains merges, separate i.e. chromatographic isolation CFIII, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios through silica gel column chromatography, portioning is collected, every 50ml collection is a, collects altogether to obtain 20～25 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 15～23 that chromatographic isolation CFIII obtains merges, through purification by silica gel column chromatography, the chloroform-methanol eluting of 10: 1 ratios obtains Hydroxy peoniflorin 0.045g;
The separation of effective ingredient in the Radix Puerariae: get Radix Puerariae 6g, with 30～80% ethanol extractions of 8 times of weight portions 3 times, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, be evaporated to 2000ml under 70～80 ℃ of conditions of temperature after, be diluted with water to 5000ml, by AB-8 type macroporous adsorptive resins, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, and collect ethanol elution, decompression and solvent recovery is to doing, residue is put 50～70 ℃ of dryings in the vacuum drying oven, obtains dry extract;
The dry extract 400g that gets method for preparing carries out silica gel column chromatography to be separated, be chromatographic isolation II, 100: 0 → 20: 1 → 10: 1 → 5: 1 → 2: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 50ml collection is a, collects altogether to obtain 80 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part the 3rd～5,6～7,8～21,22～68,69～80 that chromatographic isolation II is obtained merges respectively, obtains GA, GB, GC, GD, five parts of GE;
Part GA separates through silica gel column chromatography, be chromatographic isolation GAI, the chloroform eluting, portioning is collected, every 50ml collects a, collect altogether and obtain 10 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 2 that chromatographic isolation GAI obtains is separated through Sephadex LH-20 column chromatography, be chromatographic isolation GAII, methanol-eluted fractions, portioning is collected, and every 50ml collects a, collect altogether and obtain 8～12 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 2～5 that chromatographic isolation GAII obtains merges, and separates through silica gel column chromatography, be chromatographic isolation GAIII, the petroleum ether of 8: 1 ratios-acetone mixed solvent eluting, portioning is collected, and every 50ml collects a, collect altogether and obtain 8～12 stream parts, silica gel thin-layer chromatography is examined and is known back merging same stream part, and the stream part 4 that obtains from chromatographic isolation GAIII obtains Palmic acid 0.01g, and stream part 5 obtains arachidic acid 0.012g; Stream part 6 is separated out white crystals, uses acetone recrystallization, obtains lupeol 0.006g; Stream part 8～9 is separated out white, needle-shaped crystals, uses recrystallizing methanol, obtains cupreol 0.02g;
Part GB is separated through silica gel column chromatography, i.e. chromatographic isolation GBI, and the chloroform eluting, portioning is collected, and every 50ml collects a, collects altogether to obtain 18～24 streams part, and the silica gel thin-layer chromatography inspection is known the back and is merged same stream part; Stream part 11～17 that chromatographic isolation GBI obtains merges, separate with silica gel column chromatography, be chromatographic isolation GBII, the chloroform-methanol eluting of 20: 1 ratios, portioning is collected, and every 50ml collects a, collect altogether and obtain 18～24 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 3 that chromatographic isolation GBII obtains is separated out white precipitate, obtains lignocerane acid glyceride 0.03g behind the methanol cyclic washing; Separate out white crystals after stream part 4 placements, use recrystallizing methanol, obtain big legumin 0.035g;
Part GC separates through silica gel column chromatography, be chromatographic isolation GCI, with 20: 1 → 10: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 50ml collects a, collect altogether and obtain 60～70 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 30～42 that chromatographic isolation GCI obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation GCII, the chloroform-methanol eluting of 10: 1 ratios, portioning is collected, and every 50ml collection is a, collects altogether to obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 13～20 that chromatographic isolation GCII obtains merges, and therefrom separates out white precipitate, obtains genistin 0.02g behind the methanol cyclic washing; Stream part 43～61 that chromatographic isolation GCI obtains merges, and separates through silica gel column chromatography, i.e. chromatographic isolation GCIII, the chloroform-methanol eluting of 10: 1 ratios, portioning is collected, and every 50ml collection is a, collects altogether to obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 13～19 that chromatographic isolation GCIII obtains merges, and uses silica gel chromatography, the chloroform-methanol eluting of 10: 1 ratios, obtain 4 ', 8-dimethoxy-7-glucone isoflavone 0.025g; Separate out crystallization after stream part that chromatographic isolation GCIII obtains 21～29 is placed, use recrystallizing methanol, obtain 4 '-methoxyl group daiazi 0.045g;
Part GD dilutes with suitable quantity of water, by the AB-8 macroporous adsorbent resin, water elution to water lotion near colourless after, be washed till the eluent color with 30～70% ethanol and end when more shallow, the merging ethanol elution, decompression and solvent recovery obtains extractum 30g; Getting above-mentioned extractum separates with silica gel column chromatography, be chromatographic isolation GDI, 100: 0 → 10: 1 → 4: 1 → 2: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 100ml collects a, collect altogether and obtain 35～40 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 11～19 that chromatographic isolation GDI obtains merges, separate with silica gel column chromatography, be chromatographic isolation GDII, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios, portioning is collected, every 50ml collects a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 2～6 that chromatographic isolation GDII obtains merges, and separates with polyamide column chromatography, be chromatographic isolation GDIII, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios, portioning is collected, and every 50ml collects a, collect altogether and obtain 15～20 stream parts, the inspection of polyamide thin layer chromatography is known the back and is merged same stream part, and stream part 2～5 that chromatographic isolation GDIII obtains merges, through the polyamide column chromatography purification, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios obtains ononin 0.025g; 6～14 merge, separate through polyamide column chromatography, be chromatographic isolation GDIV, the ethyl acetate, alcohol and water mixed solvent eluting of 30: 2: 1 ratios, portioning is collected, every 50ml collects a, collect altogether and obtain 15～20 stream parts, the inspection of polyamide thin layer chromatography is known the back and is merged same stream part, stream part 3～5 that chromatographic isolation GDIV obtains merges, and polyamide column chromatography separates, i.e. chromatographic isolation GDV, the chloroform-methanol mixed solvent eluting of 15: 1 ratios, portioning is collected, and every 50ml collection is a, collects altogether to obtain 15～20 stream parts, the inspection of polyamide thin layer chromatography is known the back and is merged same stream part, after placing, stream parts 10 separates out kudzu-vine root new glycoside A 0.025g, after placing, stream parts 13～15 separates out 3 '-hydroxyl puerarin 0.01g, stream part 17～20 usefulness polyamide column chromatographies separate, the ethyl acetate, alcohol and water mixed solvent eluting of 30: 2: 1 ratios obtains daiazi 0.012g; Stream parts 14～25 merges the back to be placed, and separates out crystallization, uses recrystallizing methanol, obtain 3 '-methoxy puerarin 0.045g; Stream part 15～17 that chromatographic isolation GDIII obtains merges, and separates through polyamide column chromatography, and 7.5: 1 ratio eluting of chloroform-methanol mixed solvent obtain puerarin 0.055g; Stream part 28～36 that chromatographic isolation GDI obtains merges, separate with silica gel column chromatography, be chromatographic isolation GDVI, 5.5: 1.25: 1 ratio eluting of chloroform-methanol-water mixed solvent, portioning is collected, every 50ml collects a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, separates out crystallization after stream part 13～18 placements that chromatographic isolation GDVI obtains, use recrystallizing methanol, obtain celery glycosyl puerarin 0.05g;
The separation of effective ingredient in the Radix Ginseng: get Radix Ginseng 6g, with 30～80% ethanol extractions of 8 times of weight portions 3 times, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, be evaporated to 2000ml under 70～80 ℃ of conditions of temperature after, be diluted with water to 5000ml, by AB-8 type macroporous adsorptive resins, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, and collect ethanol elution, decompression and solvent recovery is to doing, residue is put 50～70 ℃ of dryings in the vacuum drying oven, obtains dry extract;
The dry extract 600g that gets method for preparing carries out silica gel column chromatography to be separated, be chromatographic isolation III, 19: 1～0: 1 gradient elution of chloroform-methanol mixed solvent, portioning is collected, every 100ml collection is a, collects altogether to obtain 100 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part the 16th～25,26～40,41～55,56～79,80～100 that chromatographic isolation III is obtained merges respectively, obtains RA, RB, RC, RD, five parts of RE;
Part RB separates with silica gel column chromatography, be chromatographic isolation RBI, 25: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, every 50ml collects a, collect altogether and obtain 35～40 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 14～22 merging that chromatographic isolation RBI obtains, separate with Sephadex LH-20 column chromatography, be chromatographic isolation RBII, methanol-eluted fractions, portioning is collected, every 25ml collects a, collect altogether and obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, separates out 20 (S)-ginsenoside Rhs after stream part 13～18 merging that chromatographic isolation RBII obtains 10.045g; Separate out 20 (S)-Protopanaxatriol 0.015g after stream part 20～23 merging;
Part RC separates with silica gel column chromatography, be chromatographic isolation RCI, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, every 50ml collects a, collect altogether and obtain 28～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 13～19 that chromatographic isolation RCI obtains merges, and separates with silica gel column chromatography, be chromatographic isolation RCII, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, and every 50ml collects a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 10～16 that chromatographic isolation RCII obtains merges, with SephadexLH-20 column chromatography purification, methanol-eluted fractions obtains 20 (R)-ginsenoside Rhs 10.03g;
Part RD separates with silica gel column chromatography, be chromatographic isolation RDI, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, and every 50ml collects a, collect altogether and obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 16～21 that chromatographic isolation RDI obtains merges, and separates with silica gel column chromatography, be chromatographic isolation RDII, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, and every 50ml collects a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 13～15 that chromatographic isolation RDII obtains merges, and separates through reversed-phase silica gel column chromatography, methanol-eluted fractions, portioning is collected, and every 20ml collection is a, collects altogether to obtain 15～20 stream parts, the reverse phase silica gel thin layer chromatography is examined and is known back merging same stream part, and gravity flow part 13～16 is separated out 20 (S)-ginsenoside Rgs 30.01g;
Part RE separates with silica gel column chromatography, be chromatographic isolation REI, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, and every 50ml collects a, collect altogether and obtain 83～88 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 27～38 that chromatographic isolation REI obtains merges, through the reversed-phase silica gel column chromatography purification, methanol-eluted fractions obtains the ginsenoside Rg 10.08g and 20 (S)-ginsenoside Rgs 20.03g; Stream part 42～59 merging, separate with silica gel column chromatography, be chromatographic isolation REII, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, and every 50ml collection is a, collects altogether to obtain 48～56 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 18～30 that chromatographic isolation REII obtains merges, and separates methanol-eluted fractions through reversed-phase silica gel column chromatography, portioning is collected, every 20ml collection is a, collects altogether to obtain 40～45 stream parts, and the inspection of reverse phase silica gel thin layer chromatography is known the back and merged same stream part, gravity flow part 12～18 obtains ginsenoside Rd 0.05g, and gravity flow part obtains ginsenoside Re 0.04g for 22～29 parts; Stream part 32～45 that chromatographic isolation REII obtains merges, and through the reversed-phase silica gel column chromatography purification, methanol-eluted fractions obtains ginsenoside Rb 10.06g;
Get above-mentioned each active component, mixing adds conventional adjuvant, according to common process, makes capsule.
Embodiment 12:
Radix Ginseng 3g, Radix Puerariae 18g, Radix Paeoniae Rubra 9g
The separation of effective ingredient in the Radix Paeoniae Rubra: get Radix Paeoniae Rubra 9g, with 30～80% ethanol extractions of 8 times of parts by volume 3 times, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, be evaporated to 2000ml under 70～80 ℃ of conditions of temperature after, be diluted with water to 5000ml, by AB-8 type macroporous adsorptive resins, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, and collect ethanol elution, decompression and solvent recovery is to doing, residue is put 50～70 ℃ of dryings in the vacuum drying oven, obtains dry extract;
Get the dry extract 500g of method for preparing, carry out silica gel column chromatography and separate (chromatographic isolation I), with 100: 0 → 20: 1 → 10: 1 → 5: 1 → 2: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 50ml collects a, collect altogether and obtain 115～125 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part the 4th～6,7～16,17～31,32～41,42～74,75～99,100～121 that chromatographic isolation I is obtained merges respectively, obtains CA, CB, CC, CD, CE, CF, seven different pieces of CG;
Portion C A carries out silica gel column chromatography to be separated, and promptly chromatographic isolation CAI uses 2: 1 → 7: 5 gradient elutions of petroleum ether-acetone mixed solvent, and portioning is collected, and every 50ml collection is a, and collection obtains 20～25 stream parts altogether, merges same stream part after the silica gel thin-layer chromatography inspection is known; Stream part 5～17 that chromatographic isolation CAI obtains merges, silica gel column chromatography separates, and promptly chromatographic isolation CAII uses petroleum ether-acetone mixed solvent 40～60: 1 eluting, portioning is collected, every 50ml collection is a, collects altogether to obtain 8～12 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 3～5 that chromatographic isolation CAII obtains merges, separate out the white plates crystallization, use the petroleum ether recrystallization, obtain benzoic acid 0.02g;
Portion C B carries out silica gel column chromatography to be separated, be chromatographic isolation CBI, with 100: 0 → 50: 1 → 20: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 50ml collection is a, collects altogether to obtain 30～35 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 22～27 that chromatographic isolation CBI obtains merges, separate through silica gel column chromatography, promptly chromatographic isolation CBII uses petroleum ether-acetone mixed solvent (7: 2) eluting, portioning is collected, every 50ml collection is a, collects altogether to obtain 10～15 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 9 that chromatographic isolation CBII obtains is separated through Sephadex LH-20 column chromatography, methanol-eluted fractions obtains 1S, 2S, 4R-is trans-2-hydroxyl-1, and 8-cineole 0.005g and cupreol 0.02g;
Portion C D carries out silica gel column chromatography to be separated, be chromatographic isolation CDI, petroleum ether-acetone → methanol gradient elution with the petroleum ether-acetone of 3: 1 ratios → 7.5: 5 ratio, portioning is collected, every 50ml collects a, collect altogether and obtain 75～80 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 41～70 that chromatographic isolation CDI obtains merges, and separates with silica gel column chromatography, be chromatographic isolation CDII, the petroleum ether-ethyl acetate mixed solvent eluting of 2: 1 ratios, portioning is collected, and every 50ml collects a, collect altogether and obtain 25～30 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 9～16 that chromatographic isolation CDII obtains merges, and separates through silica gel column chromatography, the petroleum ether of 5: 1 ratios-acetone eluting obtains dihydro apigenin 0.008g; The methanol-eluted fractions thing that chromatographic isolation CDI obtains separates with silica gel column chromatography, be chromatographic isolation CDIII, 2.5: the petroleum ether of 1 ratio-acetone mixed solvent eluting, portioning is collected, every 50ml collects a, collect altogether and obtain 25～30 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 9～18 that chromatographic isolation CDIII obtains merges, separate with Sephadex LH-20 column chromatography, be chromatographic isolation CDIV, the acetone eluting, portioning is collected, every 30ml collects a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 5～12 that chromatographic isolation CDIV obtains merges, and separates through silica gel column chromatography, i.e. chromatographic isolation CDV, 50: 1 → 20: 1 → 10: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, and every 50ml collection is a, collects altogether to obtain 45～50 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 8～12 that chromatographic isolation CDV obtains merges, and separates out white crystals, obtains lacdtlorin 0.03g with acetone recrystallization, stream part 13～30 merging, separate i.e. chromatographic isolation CDVI, 20: 1 → 10: 1 gradient elutions of chloroform-methanol mixed solvent through silica gel column chromatography, portioning is collected, every 50ml collection is a, collects altogether to obtain 35～40 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 4～18 that chromatographic isolation CDVI obtains merges, through the preparation of preparation of silica gel chromatographic isolation, the chloroform-methanol mixed solvent of 30: 1 ratios launches, and obtains benzoylpaeoniflorin 0.04g; Stream part 25～36 merging, separate (chromatographic isolation CDVII) through Sephadex LH-20 column chromatography, the acetone eluting, portioning is collected, and every 20ml collects a, collect altogether and obtain 10～12 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 1～3 that chromatographic isolation CDVII obtains merges, through silica gel chromatography, the chloroform-methanol mixed solvent eluting of 30: 1 ratios obtains 4-ethyl peoniflorin 0.035g; Stream part 31～42 that chromatographic isolation CDV obtains merges, and through purification by silica gel column chromatography, the chloroform-methanol eluting of 30: 1 ratios obtains 1S, 2S, 4R-anti-form-1,8-cineole-2-O-β-D-glucoside 0.04g;
Portion C E separates with silica gel column chromatography, be chromatographic isolation CEI, 7.5: 6 eluting of petroleum ether-acetone mixed solvent, portioning is collected, and every 50ml collection is a, collects altogether to obtain 28～33 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 19～30 that chromatographic isolation CEI obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation CEII, 15: 1 → 12: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, and every 50ml collection is a, collects altogether to obtain 18～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, the component 1～6 that chromatographic isolation CEII obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation CEIII, 50: 1 → 20: 1 → 8: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, and every 50ml collection is a, collects altogether to obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 22～28 usefulness Sephadex LH-20 column chromatography purification that chromatographic isolation CEIII obtains, methanol-eluted fractions obtains pyrogallol 0.006g; The component 11～12 that chromatographic isolation CEII obtains merges, separate with Sephadex LH-20 column chromatography, be chromatographic isolation CEIV, water → 30% methanol → 50% methanol gradient elution, portioning is collected, every 30ml collection is a, collects altogether to obtain 15～20 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, the component 9～14 that chromatographic isolation CEIV obtains merges, separate i.e. chromatographic isolation CEV, the chloroform-methanol of 5.5: 1.0: 0.1 ratios-water mixed solvent eluting with polyamide column chromatography, portioning is collected, every 50ml collection is a, collects altogether to obtain 15～20 stream parts, and the inspection of polyamide thin layer chromatography is known the back and merged same stream part, stream part 3～4 that chromatographic isolation CEV obtains merges, with Sephadex LH-20 column chromatography purification, methanol-eluted fractions obtains progallin A 0.01g; Stream part 8～15 merging, through the polyamide column chromatography purification, the chloroform-methanol mixed solvent eluting of 8: 1 ratios obtains gallic acid 0.025g; The component 13～15 that chromatographic isolation CEII obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation CEVI, 6: 1 eluting of chloroform-methanol mixed solvent, portioning is collected, every 50ml collection-part, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part; Stream part 14～20 that chromatographic isolation CEVI obtains merges, silica gel column chromatography separates, be chromatographic isolation CEVII, the chloroform-methanol mixed solvent eluting of 8: 1 ratios, portioning is collected, every 50ml collects a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 5～9 that chromatographic isolation CEVII obtains merges, separate with reversed-phase silica gel column chromatography, be chromatographic isolation CEVIII, 30% methanol-eluted fractions, portioning is collected, every 20ml collects a, collect altogether and obtain 15～20 stream parts, the inspection of reverse phase silica gel thin layer chromatography is known the back and is merged same stream part, and stream part 4～7 that chromatographic isolation CEVIII obtains merges, and separates with silica gel column chromatography, be chromatographic isolation CEVIX, 5.5: the chloroform-methanol of 1.0: 0.1 ratios-water mixed solvent eluting, portioning is collected, and every 50ml collects a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 10～13 that chromatographic isolation CEVIX obtains is separated through reversed-phase silica gel column chromatography, i.e. chromatographic isolation CEVX, 40% methanol-eluted fractions, portioning is collected, and every 20ml collection is a, collects altogether to obtain 15～20 stream parts, the inspection of reverse phase silica gel thin layer chromatography is known the back and is merged same stream part, stream part 14～19 that chromatographic isolation CEVX obtains is through silica gel chromatography, and the chloroform-methanol eluting of 8: 1 ratios obtains ethyl lacdtlorin A (25mg); Stream part 14～17 that chromatographic isolation CEVIX obtains merges, separate with silica gel column chromatography, be chromatographic isolation CEVXI, 5.5: the chloroform-methanol of 1.0: 0.1 ratios-water mixed solvent eluting, portioning is collected, every 50ml collects a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 14～17 merging that chromatographic isolation CEVXI obtains, through Sephadex LH-20 column chromatography purification, methanol-eluted fractions obtains (1S, 2S, 4R)-and anti-form-1,8-cineole-2-O-(6-O-α-L-rhamanopyranosyl)-β-D-glucoside 0.03g;
Portion C F separates through the Toyopearl column chromatography, be chromatographic isolation CFI, the water eluting, portioning is collected, every 20ml collects a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 5 usefulness silica gel column chromatographies that chromatographic isolation CFI obtains separate, i.e. chromatographic isolation CFII, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios, portioning is collected, every 50ml collection is a, collects altogether to obtain 20～25 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 13～19 that chromatographic isolation CFII obtains merges, use silica gel chromatography, the ethyl acetate, alcohol and water eluting of 20: 2: 1 ratios obtains peoniflorin 0.03g; Stream part 6～7 that chromatographic isolation CFI obtains merges, separate i.e. chromatographic isolation CFIII, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios through silica gel column chromatography, portioning is collected, every 50ml collection is a, collects altogether to obtain 20～25 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 15～23 that chromatographic isolation CFIII obtains merges, through purification by silica gel column chromatography, the chloroform-methanol eluting of 10: 1 ratios obtains Hydroxy peoniflorin 0.045g;
The separation of effective ingredient in the Radix Puerariae: get Radix Puerariae 18g, with 30～80% ethanol extractions of 8 times of weight portions 3 times, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, be evaporated to 2000ml under 70～80 ℃ of conditions of temperature after, be diluted with water to 5000ml, by AB-8 type macroporous adsorptive resins, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, and collect ethanol elution, decompression and solvent recovery is to doing, residue is put 50～70 ℃ of dryings in the vacuum drying oven, obtains dry extract;
The dry extract 400g that gets method for preparing carries out silica gel column chromatography to be separated, be chromatographic isolation II, 100: 0 → 20: 1 → 10: 1 → 5: 1 → 2: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 50ml collection is a, collects altogether to obtain 80 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part the 3rd～5,6～7,8～21,22～68,69～80 that chromatographic isolation II is obtained merges respectively, obtains GA, GB, GC, GD, five parts of GE;
Part GA separates through silica gel column chromatography, be chromatographic isolation GAI, the chloroform eluting, portioning is collected, every 50ml collects a, collect altogether and obtain 10 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 2 that chromatographic isolation GAI obtains is separated through Sephadex LH-20 column chromatography, be chromatographic isolation GAII, methanol-eluted fractions, portioning is collected, and every 50ml collects a, collect altogether and obtain 8～12 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 2～5 that chromatographic isolation GAII obtains merges, and separates through silica gel column chromatography, be chromatographic isolation GAIII, the petroleum ether of 8: 1 ratios-acetone mixed solvent eluting, portioning is collected, and every 50ml collects a, collect altogether and obtain 8～12 stream parts, silica gel thin-layer chromatography is examined and is known back merging same stream part, and the stream part 4 that obtains from chromatographic isolation GAIII obtains Palmic acid 0.01g, and stream part 5 obtains arachidic acid 0.012g; Stream part 6 is separated out white crystals, uses acetone recrystallization, obtains lupeol 0.006g; Stream part 8～9 is separated out white, needle-shaped crystals, uses recrystallizing methanol, obtains cupreol 0.02g;
Part GB is separated through silica gel column chromatography, be chromatographic isolation GBI, the chloroform eluting, portioning is collected, every 50ml collection is a, collects altogether to obtain 18～24 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 11～17 that chromatographic isolation GBI obtains merges, separate i.e. chromatographic isolation GBII, the chloroform-methanol eluting of 20: 1 ratios with silica gel column chromatography, portioning is collected, every 50ml collection is a, collects altogether to obtain 18～24 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 3 that chromatographic isolation GBII obtains is separated out white precipitate, obtains lignocerane acid glyceride 0.03g behind the methanol cyclic washing; Separate out white crystals after stream part 4 placements, use recrystallizing methanol, obtain big legumin 0.035g;
Part GC separates through silica gel column chromatography, be chromatographic isolation GCI, with 20: 1 → 10: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 50ml collects a, collect altogether and obtain 60～70 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 30～42 that chromatographic isolation GCI obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation GCII, the chloroform-methanol eluting of 10: 1 ratios, portioning is collected, and every 50ml collection is a, collects altogether to obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 13～20 that chromatographic isolation GCII obtains merges, and therefrom separates out white precipitate, obtains genistin 0.02g behind the methanol cyclic washing; Stream part 43～61 that chromatographic isolation GCI obtains merges, and separates through silica gel column chromatography, i.e. chromatographic isolation GCIII, the chloroform-methanol eluting of 10: 1 ratios, portioning is collected, and every 50ml collection is a, collects altogether to obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 13～19 that chromatographic isolation GCIII obtains merges, and uses silica gel chromatography, the chloroform-methanol eluting of 10: 1 ratios, obtain 4 ', 8-dimethoxy-7-glucone isoflavone 0.025g; Separate out crystallization after stream part that chromatographic isolation GCIII obtains 21～29 is placed, use recrystallizing methanol, obtain 4 '-methoxyl group daiazi 0.045g;
Part GD dilutes with suitable quantity of water, by the AB-8 macroporous adsorbent resin, water elution to water lotion near colourless after, be washed till the eluent color with 30～70% ethanol and end when more shallow, the merging ethanol elution, decompression and solvent recovery obtains extractum 30g; Getting above-mentioned extractum separates with silica gel column chromatography, be chromatographic isolation GDI, 100: 0 → 10: 1 → 4: 1 → 2: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 100ml collects a, collect altogether and obtain 35～40 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 11～19 that chromatographic isolation GDI obtains merges, separate with silica gel column chromatography, be chromatographic isolation GDII, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios, portioning is collected, every 50ml collects a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 2～6 that chromatographic isolation GDII obtains merges, and separates with polyamide column chromatography, be chromatographic isolation GDIII, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios, portioning is collected, and every 50ml collects a, collect altogether and obtain 15～20 stream parts, the inspection of polyamide thin layer chromatography is known the back and is merged same stream part, and stream part 2～5 that chromatographic isolation GDIII obtains merges, through the polyamide column chromatography purification, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios obtains ononin 0.025g; 6～14 merge, separate through polyamide column chromatography, be chromatographic isolation GDIV, the ethyl acetate, alcohol and water mixed solvent eluting of 30: 2: 1 ratios, portioning is collected, every 50ml collects a, collect altogether and obtain 15～20 stream parts, the inspection of polyamide thin layer chromatography is known the back and is merged same stream part, stream part 3～5 that chromatographic isolation GDIV obtains merges, and polyamide column chromatography separates, i.e. chromatographic isolation GDV, the chloroform-methanol mixed solvent eluting of 15: 1 ratios, portioning is collected, and every 50ml collection is a, collects altogether to obtain 15～20 stream parts, the inspection of polyamide thin layer chromatography is known the back and is merged same stream part, after placing, stream parts 10 separates out kudzu-vine root new glycoside A 0.025g, after placing, stream parts 13～15 separates out 3 '-hydroxyl puerarin 0.01g, stream part 17～20 usefulness polyamide column chromatographies separate, the ethyl acetate, alcohol and water mixed solvent eluting of 30: 2: 1 ratios obtains daiazi 0.012g; Stream parts 14～25 merges the back to be placed, and separates out crystallization, uses recrystallizing methanol, obtain 3 '-methoxy puerarin 0.045g; Stream part 15～17 that chromatographic isolation GDIII obtains merges, and separates through polyamide column chromatography, and 7.5: 1 ratio eluting of chloroform-methanol mixed solvent obtain puerarin 0.055g; Stream part 28～36 that chromatographic isolation GDI obtains merges, separate with silica gel column chromatography, be chromatographic isolation GDVI, 5.5: 1.25: 1 ratio eluting of chloroform-methanol-water mixed solvent, portioning is collected, every 50ml collects a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, separates out crystallization after stream part 13～18 placements that chromatographic isolation GDVI obtains, use recrystallizing methanol, obtain celery glycosyl puerarin 0.05g;
The separation of effective ingredient in the Radix Ginseng: get Radix Ginseng 3g, with 30～80% ethanol extractions of 8 times of weight portions 3 times, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, be evaporated to 2000ml under 70～80 ℃ of conditions of temperature after, be diluted with water to 5000ml, by AB-8 type macroporous adsorptive resins, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, and collect ethanol elution, decompression and solvent recovery is to doing, residue is put 50～70 ℃ of dryings in the vacuum drying oven, obtains dry extract;
The dry extract 600g that gets method for preparing carries out silica gel column chromatography to be separated, be chromatographic isolation III, 19: 1～0: 1 gradient elution of chloroform-methanol mixed solvent, portioning is collected, every 100ml collection is a, collects altogether to obtain 100 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part the 16th～25,26～40,41～55,56～79,80～100 that chromatographic isolation III is obtained merges respectively, obtains RA, RB, RC, RD, five parts of RE;
Part RB separates with silica gel column chromatography, be chromatographic isolation RBI, 25: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, every 50ml collects a, collect altogether and obtain 35～40 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 14～22 merging that chromatographic isolation RBI obtains, separate with Sephadex LH-20 column chromatography, be chromatographic isolation RBII, methanol-eluted fractions, portioning is collected, every 25ml collects a, collect altogether and obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, separates out 20 (S)-ginsenoside Rhs after stream part 13～18 merging that chromatographic isolation RBII obtains 10.045g; Separate out 20 (S)-Protopanaxatriol 0.015g after stream part 20～23 merging;
Part RC separates with silica gel column chromatography, be chromatographic isolation RCI, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, every 50ml collects a, collect altogether and obtain 28～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 13～19 that chromatographic isolation RCI obtains merges, and separates with silica gel column chromatography, be chromatographic isolation RCII, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, and every 50ml collects a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 10～16 that chromatographic isolation RCII obtains merges, with SephadexLH-20 column chromatography purification, methanol-eluted fractions obtains 20 (R)-ginsenoside Rhs 10.03g;
Part RD separates with silica gel column chromatography, be chromatographic isolation RDI, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, and every 50ml collects a, collect altogether and obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 16～21 that chromatographic isolation RDI obtains merges, and separates with silica gel column chromatography, be chromatographic isolation RDII, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, and every 50ml collects a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 13～15 that chromatographic isolation RDII obtains merges, and separates through reversed-phase silica gel column chromatography, methanol-eluted fractions, portioning is collected, and every 20ml collection is a, collects altogether to obtain 15～20 stream parts, the reverse phase silica gel thin layer chromatography is examined and is known back merging same stream part, and gravity flow part 13～16 is separated out 20 (S)-ginsenoside Rgs 30.01g;
Part RE separates with silica gel column chromatography, be chromatographic isolation REI, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, and every 50ml collects a, collect altogether and obtain 83～88 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 27～38 that chromatographic isolation REI obtains merges, through the reversed-phase silica gel column chromatography purification, methanol-eluted fractions obtains the ginsenoside Rg 10.08g and 20 (S)-ginsenoside Rgs 20.03g; Stream part 42～59 merging, separate with silica gel column chromatography, be chromatographic isolation REII, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, and every 50ml collection is a, collects altogether to obtain 48～56 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 18～30 that chromatographic isolation REII obtains merges, and separates methanol-eluted fractions through reversed-phase silica gel column chromatography, portioning is collected, every 20ml collection is a, collects altogether to obtain 40～45 stream parts, and the inspection of reverse phase silica gel thin layer chromatography is known the back and merged same stream part, gravity flow part 12～18 obtains ginsenoside Rd 0.05g, and gravity flow part obtains ginsenoside Re 0.04g for 22～29 parts; Stream part 32～45 that chromatographic isolation REII obtains merges, and through the reversed-phase silica gel column chromatography purification, methanol-eluted fractions obtains ginsenoside Rb 10.06g;
Above-mentioned active component mixes, and adds conventional adjuvant, according to common process, makes oral liquid.

Claims (16)

1, a kind of pharmaceutical composition of anti-angiogenic dementia is characterized in that the raw material of this pharmaceutical composition consists of: Radix Ginseng 3～18 weight portions, Radix Puerariae 3～18 weight portions, Radix Paeoniae Rubra 3～18 weight portions.
2, pharmaceutical composition as claimed in claim 1 is characterized in that the raw material of this pharmaceutical composition consists of: Radix Ginseng 10 weight portions, Radix Puerariae 10 weight portions, Radix Paeoniae Rubra 10 weight portions.
3, pharmaceutical composition as claimed in claim 1 is characterized in that the raw material of this pharmaceutical composition consists of: Radix Ginseng 3 weight portions, Radix Puerariae 18 weight portions, Radix Paeoniae Rubra 9 weight portions.
4, pharmaceutical composition as claimed in claim 1 is characterized in that the raw material of this pharmaceutical composition consists of: Radix Ginseng 18 weight portions, Radix Puerariae 3 weight portions, Radix Paeoniae Rubra 12 weight portions.
5,, it is characterized in that this method is optional following a kind of as the preparation method of the arbitrary described pharmaceutical composition effective site of claim 1-4:
A, weighting raw materials according to the above ratio, mix homogeneously, 30～80% ethanol extractions 1～4 time that add 6～10 times of parts by volume, each 1～3 hour, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, be evaporated to 1-3 times of volume of medical material amount under the condition that temperature is 70～80 ℃, relative density is about 1.04～1.05 concentrated solution under 60～65 ℃ the condition; Get concentrated solution, the water that adds 2-6 times of medical material amount stirs, adsorb by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery promptly gets pharmaceutical composition effective site of the present invention to doing;
B, take by weighing Radix Ginseng, 30～80% ethanol extractions 1～4 time that add 6～10 times of parts by volume, each 1～3 hour, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, is evaporated to 1-3 times of volume of medical material amount under the condition that temperature is 70～80 ℃, and relative density is about 1.04～1.05 concentrated solution under 60～65 ℃ the condition; Get concentrated solution, the water that adds 2-6 times of medical material amount stirs, adsorb by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery gets the Radix Ginseng extract part to doing;
Take by weighing Radix Puerariae, 30～80% ethanol extractions 1～4 time that add 6～10 times of parts by volume, each 1～3 hour, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, is evaporated to 1-3 times of volume of medical material amount under 70～80 ℃ of conditions of temperature, and relative density is about 1.04～1.05 concentrated solution under 60～65 ℃ of conditions; Get concentrated solution, the water that adds 2-6 times of medical material amount stirs, adsorb by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery gets the Radix Puerariae extract part to doing;
Take by weighing Radix Paeoniae Rubra, 30～80% ethanol extractions 1～4 time that add 6～10 times of parts by volume, each 1～3 hour, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, is evaporated to 1-3 times of volume of medical material amount under 70～80 ℃ of conditions of temperature, and 60～65 ℃ of following relative densities are about 1.04～1.05 concentrated solution; Get concentrated solution, the water that adds 2-6 times of medical material amount stirs, adsorb by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery gets the Radix Paeoniae Rubra extract part to doing;
Get above-mentioned Radix Ginseng extract part, Radix Puerariae extract part and Radix Paeoniae Rubra extract part, by 1～20: 1～20: 1～20 mixed is even, promptly gets pharmaceutical composition effective site of the present invention.
6, the preparation method of pharmaceutical composition effective site as claimed in claim 5 is characterized in that this method is optional following a kind of:
A, weighting raw materials according to the above ratio, mix homogeneously, 30～80% ethanol extractions 3 times that add 8 times of parts by volume, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, be evaporated to 2 times of volumes of medical material amount under 70～80 ℃ of conditions of temperature, relative density is about 1.04～1.05 concentrated solution under 60～65 ℃ of conditions; Get concentrated solution, the water that adds 4 times of medical material amounts stirs, adsorb by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery promptly gets pharmaceutical composition effective site of the present invention to doing;
B, take by weighing Radix Ginseng, 30～80% ethanol extractions 3 times that add 8 times of parts by volume, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, is evaporated to 2 times of volumes of medical material amount under the condition that temperature is 70～80 ℃, and relative density is about 1.04～1.05 concentrated solution under 60～65 ℃ the condition; Get concentrated solution, the water that adds 4 times of medical material amounts stirs, adsorb by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery gets the Radix Ginseng extract part to doing;
Take by weighing Radix Puerariae, 30～80% ethanol extractions 3 times that add 8 times of parts by volume, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, is evaporated to 2 times of volumes of medical material amount under 70～80 ℃ of conditions of temperature, and relative density is about 1.04～1.05 concentrated solution under 60～65 ℃ of conditions; Get concentrated solution, the water that adds 4 times of medical material amounts stirs, adsorb by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery gets the Radix Puerariae extract part to doing;
Take by weighing Radix Paeoniae Rubra, 30～80% ethanol extractions 3 times that add 8 times of parts by volume, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, is evaporated to 2 times of volumes of medical material amount under 70～80 ℃ of conditions of temperature, and 60～65 ℃ of following relative densities are about 1.04～1.05 concentrated solution; Get concentrated solution, the water that adds 4 times of medical material amounts stirs, adsorb by AB-8 type macroporous adsorbent resin, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, collect ethanol elution, decompression and solvent recovery gets the Radix Paeoniae Rubra extract part to doing;
Get above-mentioned Radix Ginseng extract part, Radix Puerariae extract part and Radix Paeoniae Rubra extract part, by 1: 1: 1,1: 20: 1,1: 1: 20,1: 10: 10 or 3: 4: 3 mixed were even, promptly get pharmaceutical composition effective site of the present invention;
Pharmaceutical composition effective site of the present invention is light brown powder, has certain moisture, and soluble in water and Diluted Alcohol is insoluble in lipotropy organic solvents such as chloroform, petroleum ether; The ferric chloride reaction positive, aluminum chloride reacting positive, hydrochloric acid-magnesium powder reaction negative, the acetic anhydride-strong sulfuric acid response positive, the alpha-Naphthol-strong sulfuric acid response positive; Mainly contain osajin, monoterpene glycosides and saponin component, wherein puerarin content is 15.0～25.0%, daiazi, 3 '-total content of methoxy puerarin and celery glycosyl puerarin is 12.0～18.0%, paeoniflorin content is 7.0～15.0%, and the content of Radix Ginseng total saponins is 17.0～25.0%.
7,, it is characterized in that this method is as the preparation method of the arbitrary described pharmaceutical composition active component of claim 1-4:
The separation of effective ingredient in the Radix Paeoniae Rubra: get Radix Paeoniae Rubra 3-18 weight portion, with 30～80% ethanol extractions of 6-10 times of parts by volume 1-4 time, each 1-3 hour, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, after being evaporated to the 1500-2500 parts by volume under 70～80 ℃ of conditions of temperature, be diluted with water to the 4000-6000 parts by volume, by AB-8 type macroporous adsorptive resins, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, and collect ethanol elution, decompression and solvent recovery is to doing, residue is put 50～70 ℃ of dryings in the vacuum drying oven, obtains dry extract;
Get the dry extract 400-600 weight portion of method for preparing, carry out silica gel column chromatography and separate (chromatographic isolation I), with 100: 0 → 20: 1 → 10: 1 → 5: 1 → 2: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 115～125 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part the 4th～6 that chromatographic isolation I is obtained, 7～16,17～31,32～41,42～74,75～99,100～121 merge respectively, obtain CA, CB, CC, CD, CE, CF, seven different pieces of CG;
Portion C A carries out silica gel column chromatography to be separated, and promptly chromatographic isolation CAI uses 2: 1 → 7: 5 gradient elutions of petroleum ether-acetone mixed solvent, portioning is collected, every 40-60 parts by volume collection is a, collects altogether to obtain 20～25 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part; Stream part 5～17 that chromatographic isolation CAI obtains merges, silica gel column chromatography separates, and promptly chromatographic isolation CAII uses petroleum ether-acetone mixed solvent 40～60: 1 eluting, portioning is collected, every 40-60 parts by volume collection is a, collects altogether to obtain 8～12 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 3～5 that chromatographic isolation CAII obtains merges, separate out the white plates crystallization, use the petroleum ether recrystallization, obtain benzoic acid 0.01-0.03 weight portion;
Portion C B carries out silica gel column chromatography to be separated, be chromatographic isolation CBI, with 100: 0 → 50: 1 → 20: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 40-60 parts by volume collection is a, collects altogether to obtain 30～35 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 22～27 that chromatographic isolation CBI obtains merges, separate through silica gel column chromatography, promptly chromatographic isolation CBII uses petroleum ether-acetone mixed solvent (7: 2) eluting, portioning is collected, every 50ml collection is a, collects altogether to obtain 10～15 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 9 that chromatographic isolation CBII obtains is separated through Sephadex LH-20 column chromatography, methanol-eluted fractions obtains 1S, 2S, 4R-is trans-2-hydroxyl-1, and 8-cineole 0.004-0.006 weight portion and cupreol 0.01-0.03 weight portion;
Portion C D carries out silica gel column chromatography to be separated, be chromatographic isolation CDI, use 2-4: the petroleum ether-acetone of 1 ratio → 5-10: the petroleum ether-acetone of 5 ratios → methanol gradient elution, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 75～80 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 41～70 that chromatographic isolation CDI obtains merges, and separates with silica gel column chromatography, be chromatographic isolation CDII, the petroleum ether-ethyl acetate mixed solvent eluting of 2: 1 ratios, portioning is collected, and every 40-60 parts by volume is collected a, collect altogether and obtain 25～30 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 9～16 that chromatographic isolation CDII obtains merges, and separates through silica gel column chromatography, 4-6: the petroleum ether of 1 ratio-acetone eluting obtains dihydro apigenin 0.006-0.01 weight portion; The methanol-eluted fractions thing that chromatographic isolation CDI obtains separates with silica gel column chromatography, be chromatographic isolation CDIII, 2-3: the petroleum ether of 1 ratio-acetone mixed solvent eluting, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 25～30 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 9～18 that chromatographic isolation CDIII obtains merges, separate with Sephadex LH-20 column chromatography, be chromatographic isolation CDIV, the acetone eluting, portioning is collected, every 20-40 parts by volume is collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 5～12 that chromatographic isolation CDIV obtains merges, separate through silica gel column chromatography, be chromatographic isolation CDV, 50: 1 → 20: 1 → 10: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 45～50 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 8～12 that chromatographic isolation CDV obtains merges, and separates out white crystals, obtains lacdtlorin 0.02-0.04 weight portion with acetone recrystallization, stream part 13～30 merging, separate i.e. chromatographic isolation CDVI, 20: 1 → 10: 1 gradient elutions of chloroform-methanol mixed solvent through silica gel column chromatography, portioning is collected, every 40-60 parts by volume collection is a, collects altogether to obtain 35～40 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 4～18 that chromatographic isolation CDVI obtains merges, through the preparation of preparation of silica gel chromatographic isolation, 20-40: the chloroform-methanol mixed solvent of 1 ratio launches, and obtains benzoylpaeoniflorin 0.03-0.05 weight portion; Stream part 25～36 merging, separate (chromatographic isolation CDVII) through Sephadex LH-20 column chromatography, the acetone eluting, portioning is collected, and every 20ml collects a, collect altogether and obtain 10～12 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 1～3 that chromatographic isolation CDVII obtains merges, through silica gel chromatography, 20-40: the chloroform-methanol mixed solvent eluting of 1 ratio obtains 4-ethyl peoniflorin 0.025-0.045 weight portion; Stream part 31～42 that chromatographic isolation CDV obtains merges, and through purification by silica gel column chromatography, 20-40: the chloroform-methanol eluting of 1 ratio obtains 1S, 2S, 4R-anti-form-1,8-cineole-2-O-β-D-glucoside 0.03-0.05 weight portion;
Portion C E separates with silica gel column chromatography, be chromatographic isolation CEI, petroleum ether-acetone mixed solvent 6-9: 6 eluting, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 28～33 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 19～30 that chromatographic isolation CEI obtains merges, separate with silica gel column chromatography, be chromatographic isolation CEII, 15: 1 → 12: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 18～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, the component 1～6 that chromatographic isolation CEII obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation CEIII, 50: 1 → 20: 1 → 8: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, and every 40-60 parts by volume collection is a, collects altogether to obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 22～28 usefulness Sephadex LH-20 column chromatography purification that chromatographic isolation CEIII obtains, methanol-eluted fractions obtains pyrogallol 0.004-0.008 weight portion; The component 11～12 that chromatographic isolation CEII obtains merges, separate with Sephadex LH-20 column chromatography, be chromatographic isolation CEIV, water → 30% methanol → 50% methanol gradient elution, portioning is collected, every 40-60 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, the component 9～14 that chromatographic isolation CEIV obtains merges, separate i.e. chromatographic isolation CEV, 4～7: 0.5～1.5: the chloroform-methanol of 0.1 ratio-water mixed solvent eluting with polyamide column chromatography, portioning is collected, every 40-60 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, and the inspection of polyamide thin layer chromatography is known the back and merged same stream part, stream part 3～4 that chromatographic isolation CEV obtains merges, with Sephadex LH-20 column chromatography purification, methanol-eluted fractions obtains progallin A 0.005-0.015 weight portion; Stream part 8～15 merging, through the polyamide column chromatography purification, 6-10: the chloroform-methanol mixed solvent eluting of 1 ratio obtains gallic acid 0.015-0.035 weight portion; The component 13～15 that chromatographic isolation CEII obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation CEVI, chloroform-methanol mixed solvent 5-7: 1 eluting, portioning is collected, and every 40-60 parts by volume is collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part; Stream part 14～20 that chromatographic isolation CEVI obtains merges, silica gel column chromatography separates, be chromatographic isolation CEVII, 6-10: the chloroform-methanol mixed solvent eluting of 1 ratio, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 5～9 that chromatographic isolation CEVII obtains merges, separate with reversed-phase silica gel column chromatography, be chromatographic isolation CEVIII, 30% methanol-eluted fractions, portioning is collected, every 10-30 parts by volume is collected a, collect altogether and obtain 15～20 stream parts, the inspection of reverse phase silica gel thin layer chromatography is known the back and is merged same stream part, stream part 4～7 that chromatographic isolation CEVIII obtains merges, separate with silica gel column chromatography, be chromatographic isolation CEVIX, 4～7: 0.5～1.5: the chloroform-methanol of 0.1 ratio-water mixed solvent eluting, portioning is collected, and every 40-60 parts by volume is collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 10～13 that chromatographic isolation CEVIX obtains is separated through reversed-phase silica gel column chromatography, i.e. chromatographic isolation CEVX, 40% methanol-eluted fractions, portioning is collected, and every 10-30 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, the inspection of reverse phase silica gel thin layer chromatography is known the back and is merged same stream part, stream part 14～19 that chromatographic isolation CEVX obtains is through silica gel chromatography, and 6-10: the chloroform-methanol eluting of 1 ratio obtains ethyl lacdtlorin A (25mg); Stream part 14～17 that chromatographic isolation CEVIX obtains merges, separate with silica gel column chromatography, be chromatographic isolation CEVXI, 4～7: 0.5～1.5: the chloroform-methanol of 0.1 ratio-water mixed solvent eluting, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 14～17 merging that chromatographic isolation CEVXI obtains, through Sephadex LH-20 column chromatography purification, methanol-eluted fractions obtains (1S, 2S, 4R)-and anti-form-1,8-cineole-2-O-(6-O-α-L-rhamanopyranosyl)-β-D-glucoside 0.02-0.04 weight portion;
Portion C F separates through the Toyopearl column chromatography, be chromatographic isolation CFI, the water eluting, portioning is collected, every 10-30 parts by volume is collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 5 usefulness silica gel column chromatographies that chromatographic isolation CFI obtains separate, i.e. chromatographic isolation CFII, 17～23: 1～3: the ethyl acetate, alcohol and water mixed solvent eluting of 1 ratio, portioning is collected, every 40-60 parts by volume collection is a, collects altogether to obtain 20～25 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 13～19 that chromatographic isolation CFII obtains merges, use silica gel chromatography, 17～23: 1～3: the ethyl acetate, alcohol and water eluting of 1 ratio obtains peoniflorin 0.02-0.04 weight portion; Stream part 6～7 that chromatographic isolation CFI obtains merges, separate through silica gel column chromatography, be chromatographic isolation CFIII, 17～23: 1～3: the ethyl acetate, alcohol and water mixed solvent eluting of 1 ratio, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 15～23 that chromatographic isolation CFIII obtains merges, through purification by silica gel column chromatography, 5-15: the chloroform-methanol eluting of 1 ratio obtains Hydroxy peoniflorin 0.035-0.055 weight portion;
The separation of effective ingredient in the Radix Puerariae: get Radix Puerariae 3-18 weight portion, with 30～80% ethanol extractions of 6-10 times of weight portion 1-4 time, each 1-3 hour, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, after being evaporated to the 1500-2500 parts by volume under 70～80 ℃ of conditions of temperature, be diluted with water to the 4000-6000 parts by volume, by AB-8 type macroporous adsorptive resins, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, and collect ethanol elution, decompression and solvent recovery is to doing, residue is put 50～70 ℃ of dryings in the vacuum drying oven, obtains dry extract;
The above-mentioned dry extract 300-500 weight portion of access method preparation carries out silica gel column chromatography to be separated, be chromatographic isolation II, 100: 0 → 20: 1 → 10: 1 → 5: 1 → 2: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 80 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part the 3rd～5,6～7,8～21,22～68,69～80 that chromatographic isolation II is obtained merges respectively, obtains GA, GB, GC, GD, five parts of GE;
Part GA separates through silica gel column chromatography, be chromatographic isolation GAI, the chloroform eluting, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 10 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 2 that chromatographic isolation GAI obtains is separated through Sephadex LH-20 column chromatography, be chromatographic isolation GAII, methanol-eluted fractions, portioning is collected, and every 40-60 parts by volume is collected a, collect altogether and obtain 8～12 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 2～5 that chromatographic isolation GAII obtains merges, and separates through silica gel column chromatography, be chromatographic isolation GAIII, 6-10: the petroleum ether of 1 ratio-acetone mixed solvent eluting, portioning is collected, and every 40-60 parts by volume is collected a, collect altogether and obtain 8～12 stream parts, silica gel thin-layer chromatography is examined and is known back merging same stream part, and the stream part 4 that obtains from chromatographic isolation GAIII obtains Palmic acid 0.005-0.015 weight portion, and stream part 5 obtains arachidic acid 0.01-0.015 weight portion; Stream part 6 is separated out white crystals, uses acetone recrystallization, obtains lupeol 0.004-0.008 weight portion; Stream part 8～9 is separated out white, needle-shaped crystals, uses recrystallizing methanol, obtains cupreol 0.01-0.03 weight portion;
Part GB is separated through silica gel column chromatography, be chromatographic isolation GBI, the chloroform eluting, portioning is collected, every 40-60 parts by volume collection is a, collects altogether to obtain 18～24 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 11～17 that chromatographic isolation GBI obtains merges, separate i.e. chromatographic isolation GBII, 10-30: the chloroform-methanol eluting of 1 ratio with silica gel column chromatography, portioning is collected, every 40-60 parts by volume collection is a, collects altogether to obtain 18～24 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 3 that chromatographic isolation GBII obtains is separated out white precipitate, obtains lignocerane acid glyceride 0.02-0.04 weight portion behind the methanol cyclic washing; Separate out white crystals after stream part 4 placements, use recrystallizing methanol, obtain big legumin 0.025-0.045 weight portion;
Part GC separates through silica gel column chromatography, be chromatographic isolation GCI, with 20: 1 → 10: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 60～70 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 30～42 that chromatographic isolation GCI obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation GCII, 5-15: the chloroform-methanol eluting of 1 ratio, portioning is collected, and every 40-60 parts by volume collection is a, collects altogether to obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 13～20 that chromatographic isolation GCII obtains merges, and therefrom separates out white precipitate, obtains genistin 0.01-0.03 weight portion behind the methanol cyclic washing; Stream part 43～61 that chromatographic isolation GCI obtains merges, separate through silica gel column chromatography, be chromatographic isolation GCIII, 5-15: the chloroform-methanol eluting of 1 ratio, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 13～19 that chromatographic isolation GCIII obtains merges, and uses silica gel chromatography, 5-15: the chloroform-methanol eluting of 1 ratio, obtain 4 ', 8-dimethoxy-7-glucone isoflavone 0.015-0.035 weight portion; Separate out crystallization after stream part that chromatographic isolation GCIII obtains 21～29 is placed, use recrystallizing methanol, obtain 4 '-methoxyl group daiazi 0.035-0.055 weight portion;
Part GD dilutes with suitable quantity of water, by the AB-8 macroporous adsorbent resin, water elution to water lotion near colourless after, be washed till the eluent color with 30～70% ethanol and end when more shallow, the merging ethanol elution, decompression and solvent recovery obtains extractum 20-40 weight portion; Getting above-mentioned extractum separates with silica gel column chromatography, be chromatographic isolation GDI, 100: 0 → 10: 1 → 4: 1 → 2: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, every 50-150 parts by volume is collected a, collect altogether and obtain 35～40 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 11～19 that chromatographic isolation GDI obtains merges, separate with silica gel column chromatography, be chromatographic isolation GDII, 17-23: 1-3: the ethyl acetate, alcohol and water mixed solvent eluting of 1 ratio, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 2～6 that chromatographic isolation GDII obtains merges, separate with polyamide column chromatography, be chromatographic isolation GDIII, 17-23: 1-3: the ethyl acetate, alcohol and water mixed solvent eluting of 1 ratio, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 15～20 stream parts, the inspection of polyamide thin layer chromatography is known the back and is merged same stream part, and stream part 2～5 that chromatographic isolation GDIII obtains merges, through the polyamide column chromatography purification, 17-23: 1-3: the ethyl acetate, alcohol and water mixed solvent eluting of 1 ratio obtains ononin 0.025 weight portion; 6～14 merge, separate through polyamide column chromatography, be chromatographic isolation GDIV, 25-35: 1-3: the ethyl acetate, alcohol and water mixed solvent eluting of 1 ratio, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 15～20 stream parts, the inspection of polyamide thin layer chromatography is known the back and is merged same stream part, stream part 3～5 that chromatographic isolation GDIV obtains merges, and polyamide column chromatography separates, i.e. chromatographic isolation GDV, 12-18: the chloroform-methanol mixed solvent eluting of 1 ratio, portioning is collected, and every 40-60 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, the inspection of polyamide thin layer chromatography is known the back and is merged same stream part, after placing, stream parts 10 separates out kudzu-vine root new glycoside A 0.015-0.035 weight portion, after placing, stream parts 13～15 separates out 3 '-hydroxyl puerarin 0.005-0.015 weight portion, stream part 17～20 usefulness polyamide column chromatographies separate, 25-35: 1-3: the ethyl acetate, alcohol and water mixed solvent eluting of 1 ratio obtains daiazi 0.01-0.015 weight portion; Stream parts 14～25 merges the back to be placed, and separates out crystallization, uses recrystallizing methanol, obtain 3 '-methoxy puerarin 0.035-0.055 weight portion; Stream part 15～17 that chromatographic isolation GDIII obtains merges, and separates through polyamide column chromatography, and chloroform-methanol mixed solvent 5-10: 1 ratio eluting obtains puerarin 0.045-0.065 weight portion; Stream part 28～36 that chromatographic isolation GDI obtains merges, separate with silica gel column chromatography, be chromatographic isolation GDVI, chloroform-methanol-water mixed solvent 3-8: 0.5-2: 1 ratio eluting, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, separates out crystallization after stream part 13～18 placements that chromatographic isolation GDVI obtains, use recrystallizing methanol, obtain celery glycosyl puerarin 0.04-0.06 weight portion;
The separation of effective ingredient in the Radix Ginseng: get Radix Ginseng 3-18 weight portion, with 30～80% ethanol extractions of 6-10 times of weight portion 1-4 time, each 1-3 hour, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, after being evaporated to the 1500-2500 parts by volume under 70～80 ℃ of conditions of temperature, be diluted with water to the 4000-6000 parts by volume, by AB-8 type macroporous adsorptive resins, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, and collect ethanol elution, decompression and solvent recovery is to doing, residue is put 50～70 ℃ of dryings in the vacuum drying oven, obtains dry extract;
The dry extract 400-800 weight portion of getting method for preparing carries out silica gel column chromatography to be separated, be chromatographic isolation III, 19: 1～0: 1 gradient elution of chloroform-methanol mixed solvent, portioning is collected, every 50-150 parts by volume collection is a, collects altogether to obtain 100 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part the 16th～25,26～40,41～55,56～79,80～100 that chromatographic isolation III is obtained merges respectively, obtains RA, RB, RC, RD, five parts of RE;
Part RB separates with silica gel column chromatography, be chromatographic isolation RBI, chloroform-methanol mixed solvent 15-35: 1 ratio eluting, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 35～40 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 14～22 merging that chromatographic isolation RBI obtains, separate with Sephadex LH-20 column chromatography, be chromatographic isolation RBII, methanol-eluted fractions, portioning is collected, every 15-35 parts by volume is collected a, collect altogether and obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, separates out 20 (S)-ginsenoside Rh1 0.035-0.055 weight portions after stream part 13～18 merging that chromatographic isolation RBII obtains; Separate out 20 (S)-Protopanaxatriol 0.005-0.025 weight portions after stream part 20～23 merging;
Part RC separates with silica gel column chromatography, be chromatographic isolation RCI, chloroform-methanol mixed solvent 5-15: 1 ratio eluting, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 28～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 13～19 that chromatographic isolation RCI obtains merges, and separates with silica gel column chromatography, be chromatographic isolation RCII, chloroform-methanol mixed solvent 5-15: 1 ratio eluting, portioning is collected, and every 40-60 parts by volume is collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 10～16 that chromatographic isolation RCII obtains merges, with Sephadex LH-20 column chromatography purification, methanol-eluted fractions obtains 20 (R)-ginsenoside Rh1 0.02-0.04 weight portions;
Part RD separates with silica gel column chromatography, be chromatographic isolation RDI, chloroform-methanol mixed solvent 5-15: 1 ratio eluting, portioning is collected, every 50ml collects a, collect altogether and obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 16～21 that chromatographic isolation RDI obtains merges, and separates with silica gel column chromatography, be chromatographic isolation RDII, chloroform-methanol mixed solvent 5-15: 1 ratio eluting, portioning is collected, and every 40-60 parts by volume is collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 13～15 that chromatographic isolation RDII obtains merges, and separates through reversed-phase silica gel column chromatography, methanol-eluted fractions, portioning is collected, and every 10-30 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, the reverse phase silica gel thin layer chromatography is examined and is known back merging same stream part, and gravity flow part 13～16 is separated out 20 (S)-ginsenoside Rg3 0.005-0.015 weight portions;
Part RE separates with silica gel column chromatography, be chromatographic isolation REI, chloroform-methanol mixed solvent 5-15: 1 ratio eluting, portioning is collected, and every 40-60 parts by volume is collected a, collect altogether and obtain 83～88 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 27～38 that chromatographic isolation REI obtains merges, through the reversed-phase silica gel column chromatography purification, methanol-eluted fractions obtains ginsenoside Rg1 0.06-0.10 weight portion and 20 (S)-ginsenoside Rg2 0.02-0.04 weight portion; Stream part 42～59 merging, separate with silica gel column chromatography, be chromatographic isolation REII, chloroform-methanol mixed solvent 5-15: 1 ratio eluting, portioning is collected, every 40-60 parts by volume is collected a, collect altogether and obtain 48～56 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 18～30 that chromatographic isolation REII obtains merges, and separates methanol-eluted fractions through reversed-phase silica gel column chromatography, portioning is collected, every 10-30 parts by volume collection is a, collects altogether to obtain 40～45 stream parts, and the inspection of reverse phase silica gel thin layer chromatography is known the back and merged same stream part, gravity flow part 12～18 obtains ginsenoside Rd 0.04-0.06 weight portion, and gravity flow part obtains ginsenoside Re 0.03-0.05 weight portion for 22～29 parts; Stream part 32～45 that chromatographic isolation REII obtains merges, and through the reversed-phase silica gel column chromatography purification, methanol-eluted fractions obtains ginsenoside Rb1 0.04-0.08 weight portion.
8, the preparation method of pharmaceutical composition active component as claimed in claim 7 is characterized in that this method is:
The separation of effective ingredient in the Radix Paeoniae Rubra: get Radix Paeoniae Rubra 6 weight portions, with 30～80% ethanol extractions of 8 times of parts by volume 3 times, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, after being evaporated to 2000 parts by volume under 70～80 ℃ of conditions of temperature, be diluted with water to 5000 parts by volume, by AB-8 type macroporous adsorptive resins, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, and collect ethanol elution, decompression and solvent recovery is to doing, residue is put 50～70 ℃ of dryings in the vacuum drying oven, obtains dry extract;
Above-mentioned dry extract 500 weight portions of access method preparation, carry out silica gel column chromatography and separate (chromatographic isolation I), with 100: 0 → 20: 1 → 10: 1 → 5: 1 → 2: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 115～125 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part the 4th～6,7～16,17～31,32～41,42～74,75～99,100～121 that chromatographic isolation I is obtained merges respectively, obtains CA, CB, CC, CD, CE, CF, seven different pieces of CG;
Portion C A carries out silica gel column chromatography to be separated, and promptly chromatographic isolation CAI uses 2: 1 → 7: 5 gradient elutions of petroleum ether-acetone mixed solvent, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 20～25 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part; Stream part 5～17 that chromatographic isolation CAI obtains merges, silica gel column chromatography separates, and promptly chromatographic isolation CAII uses petroleum ether-acetone mixed solvent 40～6: 1 eluting, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 8～12 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 3～5 that chromatographic isolation CAII obtains merges, separate out the white plates crystallization, use the petroleum ether recrystallization, obtain benzoic acid 0.02 weight portion;
Portion C B carries out silica gel column chromatography to be separated, be chromatographic isolation CBI, with 100: 0 → 50: 1 → 20: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 30～35 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 22～27 that chromatographic isolation CBI obtains merges, separate through silica gel column chromatography, promptly chromatographic isolation CBII uses petroleum ether-acetone mixed solvent (7: 2) eluting, portioning is collected, every 50ml collection is a, collects altogether to obtain 10～15 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 9 that chromatographic isolation CBII obtains is separated through Sephadex LH-20 column chromatography, methanol-eluted fractions obtains 1S, 2S, 4R-is trans-2-hydroxyl-1, and 8-cineole 0.005 weight portion and cupreol 0.02 weight portion;
Portion C D carries out silica gel column chromatography to be separated, be chromatographic isolation CDI, petroleum ether-acetone → methanol gradient elution with the petroleum ether-acetone of 3: 1 ratios → 7.5: 5 ratio, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 75～80 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 41～70 that chromatographic isolation CDI obtains merges, and separates with silica gel column chromatography, be chromatographic isolation CDII, the petroleum ether-ethyl acetate mixed solvent eluting of 2: 1 ratios, portioning is collected, and per 50 parts by volume are collected a, collect altogether and obtain 25～30 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 9～16 that chromatographic isolation CDII obtains merges, and separates through silica gel column chromatography, the petroleum ether of 5: 1 ratios-acetone eluting obtains dihydro apigenin 0.008 weight portion; The methanol-eluted fractions thing that chromatographic isolation CDI obtains separates with silica gel column chromatography, be chromatographic isolation CDIII, 2.5: the petroleum ether of 1 ratio-acetone mixed solvent eluting, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 25～30 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 9～18 that chromatographic isolation CDIII obtains merges, separate with Sephadex LH-20 column chromatography, be chromatographic isolation CDIV, the acetone eluting, portioning is collected, per 30 parts by volume are collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 5～12 that chromatographic isolation CDIV obtains merges, and separates through silica gel column chromatography, i.e. chromatographic isolation CDV, 50: 1 → 20: 1 → 10: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, and per 50 parts by volume collection is a, collects altogether to obtain 45～50 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 8～12 that chromatographic isolation CDV obtains merges, and separates out white crystals, obtains lacdtlorin 0.03 weight portion with acetone recrystallization, stream part 13～30 merging, separate i.e. chromatographic isolation CDVI, 20: 1 → 10: 1 gradient elutions of chloroform-methanol mixed solvent through silica gel column chromatography, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 35～40 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 4～18 that chromatographic isolation CDVI obtains merges, through the preparation of preparation of silica gel chromatographic isolation, the chloroform-methanol mixed solvent of 30: 1 ratios launches, and obtains benzoylpaeoniflorin 0.04 weight portion; Stream part 25～36 merging, separate (chromatographic isolation CDVII) through Sephadex LH-20 column chromatography, the acetone eluting, portioning is collected, and every 20ml collects a, collect altogether and obtain 10～12 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 1～3 that chromatographic isolation CDVII obtains merges, through silica gel chromatography, the chloroform-methanol mixed solvent eluting of 30: 1 ratios obtains 4-ethyl peoniflorin 0.035 weight portion; Stream part 31～42 that chromatographic isolation CDV obtains merges, and through purification by silica gel column chromatography, the chloroform-methanol eluting of 30: 1 ratios obtains 1S, 2S, 4R-anti-form-1,8-cineole-2-O-β-D-glucoside 0.04 weight portion;
Portion C E separates with silica gel column chromatography, be chromatographic isolation CEI, 7.5: 6 eluting of petroleum ether-acetone mixed solvent, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 28～33 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 19～30 that chromatographic isolation CEI obtains merges, separate with silica gel column chromatography, be chromatographic isolation CEII, 15: 1 → 12: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 18～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, the component 1～6 that chromatographic isolation CEII obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation CEIII, 50: 1 → 20: 1 → 8: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, and per 50 parts by volume collection is a, collects altogether to obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 22～28 usefulness Sephadex LH-20 column chromatography purification that chromatographic isolation CEIII obtains, methanol-eluted fractions obtains pyrogallol 0.006 weight portion; The component 11～12 that chromatographic isolation CEII obtains merges, separate with Sephadex LH-20 column chromatography, be chromatographic isolation CEIV, water → 30% methanol → 50% methanol gradient elution, portioning is collected, per 30 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, the component 9～14 that chromatographic isolation CEIV obtains merges, separate i.e. chromatographic isolation CEV, the chloroform-methanol of 5.5: 1.0: 0.1 ratios-water mixed solvent eluting with polyamide column chromatography, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, and the inspection of polyamide thin layer chromatography is known the back and merged same stream part, stream part 3～4 that chromatographic isolation CEV obtains merges, with Sephadex LH-20 column chromatography purification, methanol-eluted fractions obtains progallin A 0.01 weight portion; Stream part 8～15 merging, through the polyamide column chromatography purification, the chloroform-methanol mixed solvent eluting of 8: 1 ratios obtains gallic acid 0.025 weight portion; The component 13～15 that chromatographic isolation CEII obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation CEVI, 6: 1 eluting of chloroform-methanol mixed solvent, portioning is collected, and per 50 parts by volume are collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part; Stream part 14～20 that chromatographic isolation CEVI obtains merges, silica gel column chromatography separates, be chromatographic isolation CEVII, the chloroform-methanol mixed solvent eluting of 8: 1 ratios, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 5～9 that chromatographic isolation CEVII obtains merges, separate with reversed-phase silica gel column chromatography, be chromatographic isolation CEVIII, 30% methanol-eluted fractions, portioning is collected, per 20 parts by volume are collected a, collect altogether and obtain 15～20 stream parts, the inspection of reverse phase silica gel thin layer chromatography is known the back and is merged same stream part, stream part 4～7 that chromatographic isolation CEVIII obtains merges, separate with silica gel column chromatography, be chromatographic isolation CEVIX, 5.5: the chloroform-methanol of 1.0: 0.1 ratios-water mixed solvent eluting, portioning is collected, and per 50 parts by volume are collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 10～13 that chromatographic isolation CEVIX obtains is separated through reversed-phase silica gel column chromatography, i.e. chromatographic isolation CEVX, 40% methanol-eluted fractions, portioning is collected, and per 20 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, the inspection of reverse phase silica gel thin layer chromatography is known the back and is merged same stream part, stream part 14～19 that chromatographic isolation CEVX obtains is through silica gel chromatography, and the chloroform-methanol eluting of 8: 1 ratios obtains ethyl lacdtlorin A (25mg); Stream part 14～17 that chromatographic isolation CEVIX obtains merges, separate with silica gel column chromatography, be chromatographic isolation CEVXI, 5.5: the chloroform-methanol of 1.0: 0.1 ratios-water mixed solvent eluting, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 14～17 merging that chromatographic isolation CEVXI obtains, through Sephadex LH-20 column chromatography purification, methanol-eluted fractions obtains (1S, 2S, 4R)-and anti-form-1,8-cineole-2-O-(6-O-α-L-rhamanopyranosyl)-β-D-glucoside 0.03 weight portion;
Portion C F separates through the Toyopearl column chromatography, be chromatographic isolation CFI, the water eluting, portioning is collected, per 20 parts by volume are collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 5 usefulness silica gel column chromatographies that chromatographic isolation CFI obtains separate, i.e. chromatographic isolation CFII, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 20～25 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 13～19 that chromatographic isolation CFII obtains merges, use silica gel chromatography, the ethyl acetate, alcohol and water eluting of 20: 2: 1 ratios obtains peoniflorin 0.03 weight portion; Stream part 6～7 that chromatographic isolation CFI obtains merges, separate i.e. chromatographic isolation CFIII, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios through silica gel column chromatography, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 20～25 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 15～23 that chromatographic isolation CFIII obtains merges, through purification by silica gel column chromatography, the chloroform-methanol eluting of 10: 1 ratios obtains Hydroxy peoniflorin 0.045 weight portion;
The separation of effective ingredient in the Radix Puerariae: get Radix Puerariae 6 weight portions, with 30～80% ethanol extractions of 8 times of weight portions 3 times, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, after being evaporated to 2000 parts by volume under 70～80 ℃ of conditions of temperature, be diluted with water to 5000 parts by volume, by AB-8 type macroporous adsorptive resins, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, and collect ethanol elution, decompression and solvent recovery is to doing, residue is put 50～70 ℃ of dryings in the vacuum drying oven, obtains dry extract;
Above-mentioned dry extract 400 weight portions of access method preparation carry out silica gel column chromatography to be separated, be chromatographic isolation II, 100: 0 → 20: 1 → 10: 1 → 5: 1 → 2: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 80 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part the 3rd～5,6～7,8～21,22～68,69～80 that chromatographic isolation II is obtained merges respectively, obtains GA, GB, GC, GD, five parts of GE;
Part GA separates through silica gel column chromatography, be chromatographic isolation GAI, the chloroform eluting, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 10 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 2 that chromatographic isolation GAI obtains is separated through Sephadex LH-20 column chromatography, be chromatographic isolation GAII, methanol-eluted fractions, portioning is collected, and per 50 parts by volume are collected a, collect altogether and obtain 8～12 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 2～5 that chromatographic isolation GAII obtains merges, and separates through silica gel column chromatography, be chromatographic isolation GAIII, the petroleum ether of 8: 1 ratios-acetone mixed solvent eluting, portioning is collected, and per 50 parts by volume are collected a, collect altogether and obtain 8～12 stream parts, silica gel thin-layer chromatography is examined and is known back merging same stream part, and the stream part 4 that obtains from chromatographic isolation GAIII obtains Palmic acid 0.01 weight portion, and stream part 5 obtains arachidic acid 0.012 weight portion; Stream part 6 is separated out white crystals, uses acetone recrystallization, obtains lupeol 0.006 weight portion; Stream part 8～9 is separated out white, needle-shaped crystals, uses recrystallizing methanol, obtains cupreol 0.02 weight portion;
Part GB is separated through silica gel column chromatography, be chromatographic isolation GBI, the chloroform eluting, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 18～24 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 11～17 that chromatographic isolation GBI obtains merges, separate i.e. chromatographic isolation GBII, the chloroform-methanol eluting of 20: 1 ratios with silica gel column chromatography, portioning is collected, per 50 parts by volume collection is a, collects altogether to obtain 18～24 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part 3 that chromatographic isolation GBII obtains is separated out white precipitate, obtains lignocerane acid glyceride 0.03 weight portion behind the methanol cyclic washing; Separate out white crystals after stream part 4 placements, use recrystallizing methanol, obtain big legumin 0.035 weight portion;
Part GC separates through silica gel column chromatography, be chromatographic isolation GCI, with 20: 1 → 10: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 60～70 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 30～42 that chromatographic isolation GCI obtains merges, and separates with silica gel column chromatography, i.e. chromatographic isolation GCII, the chloroform-methanol eluting of 10: 1 ratios, portioning is collected, and per 50 parts by volume collection is a, collects altogether to obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 13～20 that chromatographic isolation GCII obtains merges, and therefrom separates out white precipitate, obtains genistin 0.02 weight portion behind the methanol cyclic washing; Stream part 43～61 that chromatographic isolation GCI obtains merges, and separates through silica gel column chromatography, i.e. chromatographic isolation GCIII, the chloroform-methanol eluting of 10: 1 ratios, portioning is collected, and per 50 parts by volume collection is a, collects altogether to obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 13～19 that chromatographic isolation GCIII obtains merges, and uses silica gel chromatography, the chloroform-methanol eluting of 10: 1 ratios, obtain 4 ', 8-dimethoxy-7-glucone isoflavone 0.025 weight portion; Separate out crystallization after stream part that chromatographic isolation GCIII obtains 21～29 is placed, use recrystallizing methanol, obtain 4 '-methoxyl group daiazi 0.045 weight portion;
Part GD dilutes with suitable quantity of water, by the AB-8 macroporous adsorbent resin, water elution to water lotion near colourless after, be washed till the eluent color with 30～70% ethanol and end when more shallow, the merging ethanol elution, decompression and solvent recovery obtains extractum 30 weight portions; Getting above-mentioned extractum separates with silica gel column chromatography, be chromatographic isolation GDI, 100: 0 → 10: 1 → 4: 1 → 2: 1 gradient elutions of chloroform-methanol mixed solvent, portioning is collected, per 100 parts by volume are collected a, collect altogether and obtain 35～40 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 11～19 that chromatographic isolation GDI obtains merges, separate with silica gel column chromatography, be chromatographic isolation GDII, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 2～6 that chromatographic isolation GDII obtains merges, and separates with polyamide column chromatography, be chromatographic isolation GDIII, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios, portioning is collected, and per 50 parts by volume are collected a, collect altogether and obtain 15～20 stream parts, the inspection of polyamide thin layer chromatography is known the back and is merged same stream part, and stream part 2～5 that chromatographic isolation GDIII obtains merges, through the polyamide column chromatography purification, the ethyl acetate, alcohol and water mixed solvent eluting of 20: 2: 1 ratios obtains ononin 0.025 weight portion; 6～14 merge, separate through polyamide column chromatography, be chromatographic isolation GDIV, the ethyl acetate, alcohol and water mixed solvent eluting of 30: 2: 1 ratios, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 15～20 stream parts, the inspection of polyamide thin layer chromatography is known the back and is merged same stream part, stream part 3～5 that chromatographic isolation GDIV obtains merges, and polyamide column chromatography separates, i.e. chromatographic isolation GDV, the chloroform-methanol mixed solvent eluting of 15: 1 ratios, portioning is collected, and per 50 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, the inspection of polyamide thin layer chromatography is known the back and is merged same stream part, after placing, stream parts 10 separates out kudzu-vine root new glycoside A 0.025 weight portion, after placing, stream parts 13～15 separates out 3 '-hydroxyl puerarin 0.01 weight portion, stream part 17～20 usefulness polyamide column chromatographies separate, the ethyl acetate, alcohol and water mixed solvent eluting of 30: 2: 1 ratios obtains daiazi 0.012 weight portion; Stream parts 14～25 merges the back to be placed, and separates out crystallization, uses recrystallizing methanol, obtain 3 '-methoxy puerarin 0.045 weight portion; Stream part 15～17 that chromatographic isolation GDIII obtains merges, and separates through polyamide column chromatography, and 7.5: 1 ratio eluting of chloroform-methanol mixed solvent obtain puerarin 0.055 weight portion; Stream part 28～36 that chromatographic isolation GDI obtains merges, separate with silica gel column chromatography, be chromatographic isolation GDVI, 5.5: 1.25: 1 ratio eluting of chloroform-methanol-water mixed solvent, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 15～20 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, separates out crystallization after stream part 13～18 placements that chromatographic isolation GDVI obtains, use recrystallizing methanol, obtain celery glycosyl puerarin 0.05 weight portion;
The separation of effective ingredient in the Radix Ginseng: get Radix Ginseng 6 weight portions, with 30～80% ethanol extractions of 8 times of weight portions 3 times, each 2 hours, merge each time extracting solution, at vacuum 〉=0.08Mpa, steam pressure is no more than 0.1Mpa, after being evaporated to 2000 parts by volume under 70～80 ℃ of conditions of temperature, be diluted with water to 5000 parts by volume, by AB-8 type macroporous adsorptive resins, liquid to be extracted is all by behind the resin column, and water continues the flushing resin column, to nearly colourless the ending of water lotion, and then the material that adsorbs on the resin column is carried out eluting with 30～80% ethanol, and collect ethanol elution, decompression and solvent recovery is to doing, residue is put 50～70 ℃ of dryings in the vacuum drying oven, obtains dry extract;
Dry extract 600 weight portions of getting method for preparing carry out silica gel column chromatography to be separated, be chromatographic isolation III, 19: 1～0: 1 gradient elution of chloroform-methanol mixed solvent, portioning is collected, per 100 parts by volume collection is a, collects altogether to obtain 100 stream parts, and the silica gel thin-layer chromatography inspection is known the back and merged same stream part, stream part the 16th～25,26～40,41～55,56～79,80～100 that chromatographic isolation III is obtained merges respectively, obtains RA, RB, RC, RD, five parts of RE;
Part RB separates with silica gel column chromatography, be chromatographic isolation RBI, 25: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 35～40 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 14～22 merging that chromatographic isolation RBI obtains, separate with Sephadex LH-20 column chromatography, be chromatographic isolation RBII, methanol-eluted fractions, portioning is collected, per 25 parts by volume are collected a, collect altogether and obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, separates out 20 (S)-ginsenoside Rh1s, 0.045 weight portion after stream part 13～18 merging that chromatographic isolation RBII obtains; Separate out 20 (S)-Protopanaxatriols, 0.015 weight portion after stream part 20～23 merging;
Part RC separates with silica gel column chromatography, be chromatographic isolation RCI, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, per 50 parts by volume are collected a, collect altogether and obtain 28～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 13～19 that chromatographic isolation RCI obtains merges, and separates with silica gel column chromatography, be chromatographic isolation RCII, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, and per 50 parts by volume are collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 10～16 that chromatographic isolation RCII obtains merges, with SephadexLH-20 column chromatography purification, methanol-eluted fractions obtains 20 (R)-ginsenoside Rh1s, 0.03 weight portion;
Part RD separates with silica gel column chromatography, be chromatographic isolation RDI, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, every 50ml collects a, collect altogether and obtain 30～35 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 16～21 that chromatographic isolation RDI obtains merges, and separates with silica gel column chromatography, be chromatographic isolation RDII, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, and per 50 parts by volume are collected a, collect altogether and obtain 20～25 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 13～15 that chromatographic isolation RDII obtains merges, and separates through reversed-phase silica gel column chromatography, methanol-eluted fractions, portioning is collected, and per 20 parts by volume collection is a, collects altogether to obtain 15～20 stream parts, the reverse phase silica gel thin layer chromatography is examined and is known back merging same stream part, and gravity flow part 13～16 is separated out 20 (S)-ginsenoside Rgs 30.01 weight portion;
Part RE separates with silica gel column chromatography, be chromatographic isolation REI, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, and per 50 parts by volume are collected a, collect altogether and obtain 83～88 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, and stream part 27～38 that chromatographic isolation REI obtains merges, through the reversed-phase silica gel column chromatography purification, methanol-eluted fractions obtains the ginsenoside Rg 10.08 weight portion and 20 (S)-ginsenoside Rg 20.03 weight portion; Stream part 42～59 merging, separate with silica gel column chromatography, be chromatographic isolation REII, 10: 1 ratio eluting of chloroform-methanol mixed solvent, portioning is collected, and per 50 parts by volume collection is a, collects altogether to obtain 48～56 stream parts, the silica gel thin-layer chromatography inspection is known the back and is merged same stream part, stream part 18～30 that chromatographic isolation REII obtains merges, and separates methanol-eluted fractions through reversed-phase silica gel column chromatography, portioning is collected, per 20 parts by volume collection is a, collects altogether to obtain 40～45 stream parts, and the inspection of reverse phase silica gel thin layer chromatography is known the back and merged same stream part, gravity flow part 12～18 obtains ginsenoside Rd's 0.05 weight portion, and gravity flow part obtains ginsenoside Re's 0.04 weight portion for 22～29 parts; Stream part 32～45 that chromatographic isolation REII obtains merges, and through the reversed-phase silica gel column chromatography purification, methanol-eluted fractions obtains ginsenoside Rb 10.06 weight portion.
9, as the arbitrary described pharmaceutical composition of claim 1-4, it is characterized in that technology routinely, add conventional adjuvant, make concentrated pill, capsule, drop pill, granule, tablet, soft capsule, slow releasing agent, oral liquid or the lyophilized injectable powder of clinical acceptance.
10, pharmaceutical composition effective site as claimed in claim 5, it is characterized in that technology routinely, add conventional adjuvant, make concentrated pill, capsule, drop pill, granule, tablet, soft capsule, slow releasing agent, oral liquid or the lyophilized injectable powder of clinical acceptance.
11, pharmaceutical composition active component as claimed in claim 7, it is characterized in that technology routinely, add conventional adjuvant, make concentrated pill, capsule, drop pill, granule, tablet, soft capsule, slow releasing agent, oral liquid or the lyophilized injectable powder of clinical acceptance.
12, as the application of the arbitrary described pharmaceutical composition of claim 1-4 in the anti-angiogenic dementia medicine of preparation treatment.
13, the application of pharmaceutical composition effective site as claimed in claim 5 in the anti-angiogenic dementia medicine of preparation treatment.
14, the application of pharmaceutical composition effective site as claimed in claim 6 in the anti-angiogenic dementia medicine of preparation treatment.
15, the application of pharmaceutical composition active component as claimed in claim 7 in the anti-angiogenic dementia medicine of preparation treatment.
16, the application of pharmaceutical composition active component as claimed in claim 8 in the anti-angiogenic dementia medicine of preparation treatment.
CN 200610113835 2006-10-18 2006-10-18 Preparing method for a kind of medicinal composition and its effective parts and active ingredients Expired - Fee Related CN1943642B (en)

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CN103558330A (en) * 2013-10-08 2014-02-05 九芝堂股份有限公司 Thin-layer identification chromatography method of stomach-strengthening ulcer-curing preparation
CN104072550A (en) * 2013-03-25 2014-10-01 河北以岭医药研究院有限公司 Separation method of monoterpene and saponins components in traditional Chinese medicine composition vegetable drug midbody
CN109602795A (en) * 2018-12-25 2019-04-12 北京鑫开元医药科技有限公司 A kind of pharmaceutical composition and its extracting method, pharmaceutical preparation, purposes

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CN104072550A (en) * 2013-03-25 2014-10-01 河北以岭医药研究院有限公司 Separation method of monoterpene and saponins components in traditional Chinese medicine composition vegetable drug midbody
CN104072550B (en) * 2013-03-25 2018-05-29 河北以岭医药研究院有限公司 The separation method of monoterpene and saponin component in Chinese medicine composition autonomic drug intermediate
CN103263462A (en) * 2013-06-08 2013-08-28 贵州大学 Desmodium caudatum extractive and extraction method and new application thereof
CN103263462B (en) * 2013-06-08 2014-10-15 贵州大学 Desmodium caudatum extractive and extraction method and new application thereof
CN103558330A (en) * 2013-10-08 2014-02-05 九芝堂股份有限公司 Thin-layer identification chromatography method of stomach-strengthening ulcer-curing preparation
CN103558330B (en) * 2013-10-08 2016-01-20 九芝堂股份有限公司 A kind of TLC distinguish chromatographic process of Jianwei Yuyang preparation
CN109602795A (en) * 2018-12-25 2019-04-12 北京鑫开元医药科技有限公司 A kind of pharmaceutical composition and its extracting method, pharmaceutical preparation, purposes

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