CN101474196B - Flavone active site of Vernonia cinerea as well as preparation method and use thereof - Google Patents

Flavone active site of Vernonia cinerea as well as preparation method and use thereof Download PDF

Info

Publication number
CN101474196B
CN101474196B CN2009100250805A CN200910025080A CN101474196B CN 101474196 B CN101474196 B CN 101474196B CN 2009100250805 A CN2009100250805 A CN 2009100250805A CN 200910025080 A CN200910025080 A CN 200910025080A CN 101474196 B CN101474196 B CN 101474196B
Authority
CN
China
Prior art keywords
extract
extraction
acetic acid
herba vernoniae
ethyl acetate
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related
Application number
CN2009100250805A
Other languages
Chinese (zh)
Other versions
CN101474196A (en
Inventor
朱华旭
潘林梅
唐于平
段金廒
张启春
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nanjing University of Chinese Medicine
Original Assignee
Nanjing University of Chinese Medicine
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Nanjing University of Chinese Medicine filed Critical Nanjing University of Chinese Medicine
Priority to CN2009100250805A priority Critical patent/CN101474196B/en
Publication of CN101474196A publication Critical patent/CN101474196A/en
Application granted granted Critical
Publication of CN101474196B publication Critical patent/CN101474196B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Landscapes

  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

The invention relates to a flavonoid active site separated from herb or roots of ironweed vernonia cinerea and a preparation method thereof. The flavonoid active site is separated from herbs of vernonia cinerea by chromatographic technique, comprises seven flavonoid compounds of celereoin, chrysoerial, luteolin, chrysoerial-7-O-beta-D-glucoside, luteolin-7-O-beta-D-glucoside, quercetin and celereoin-4'-O-beta-D-glucoside, and has senile dementia-resisting function.

Description

Flavonoid active site of Herba Vernoniae Cinereae and its production and application
One, technical field
The invention belongs to the Natural Medicine Chemistry technical field, be specifically related to a kind of chromatographic separation technology that utilizes and from ironweed Herba Vernoniae Cinereae herb, extract flavonoid active site and preparation technology and the application that obtains.
Two, background technology
Herba Vernoniae Cinereae is herb or the root of ironweed (Vernonia Schreb.) plant Herba Vernoniae Cinereae (Vernonia cinerea (L.) Less.[V.abbreviana (Wall.) DC.]), have another name called Herba Vernoniae Cinereae, ipomoea bonanox, star is wiped away grass " south of the Five Ridges gather medicinal herbs record ", post color grass " Guangzhou flora ", Herba Senecionis cannabifolii " Guangdong Chinese medicine ", XIAOSHANHU (Guangzhou army " Chinese herbal medicine handbook commonly used), false one-tenth shrimp, the branch Rhizoma et radix valerianae (Guangzhou air force " Chinese herbal medicine handbook commonly used), Flos Carthami Herba Veronicae " Huiyang, Guangdong Chinese herbal medicine ", four eyed grass " Wuzhou Chinese herbal medicine ", it red grass " North Sea, Guangxi traditional herbal medicine ", cane ginseng (Yunnan); This plant is an annual herb, high 20-80cm; Be born in hillside, wilderness, limit, field, roadside or thick forest, the shrubbery; Be distributed in ground such as Zhejiang, Jiangxi, Fujian, Taiwan, Hubei, Hunan, Guangdong, Hainan, Guangxi, Sichuan, Guizhou, Yunnan, Tibet.Bitter in the mouth, suffering, cool in nature; Has wind and heat dispersing, dehumidifying, antidotal effect.Cure mainly fever caused by exogenous pathogens, cough, acute icterohepatitis, damp-heat diarrhea, leucorrhea, furuncle swelling toxin, mastitis, rhinitis and venom.
To the Herba Vernoniae Cinereae The Chemical Constituents mainly is by the column chromatography chromatogram method at present, separates obtaining monomeric compound in Herba Vernoniae Cinereae, and by the definite structure of getting chemical compound of spectrographic method.As: in the herb of Herba Vernoniae Cinereae, get diosmetin (Diosmetin), luteolin-7-O-glucuronide (Luteolin-7-O-glucuronide), luteolin (Luteolin) and luteolin-7-O-glucoside (Luteolin-7-O-glucoside).Make a fresh start and get luteolin, luteolin-7-O-glucoside, isorientin (Isoorientin) and golden eriodictyol (Chrysoeriol) in the flower.In root, get 5,17 (20)-bean steroid diene-3 β-alcohol (Stigmast-5,17 (20)-dien-3 beta-ols), 26-methyl 27 carbonic acid (26-Methylheptacosanoic acid), stigmasterol (Stigmasterol), sitosterol (Sitosterol), α-, β-Amyrin (α-, β-Amyrin), δ-balsam acetas (δ-Amyrin acetate), α-, β-balsam acetas (α-, β-Amyrin acetate), 3 β-acetoxyl group-13 (18)-Usu alkene (3 β-Acetoxyurs-13 (18)-ene) and 24-hydroxyl-14-taraxerene (24-Hydroxytaraxer-14-ene).Get 8 α-Fructus Crotonis acyloxy-bristle Ramulus Uncariae Cum Uncis lactone-13-O-acetas (8 α-Tigloyloxyhirsutinolide-13-O-acetate) from aerial parts, 8 α-(hydroxyl isobutene. acyl-oxygen base-bristle Ramulus Uncariae Cum Uncis lactone-13-acetas (8 α-(Hydroxymethacryloyloxy)-hirsutinolide-13-O-acetate), the appropriate graceful lactone of this terraced promise-8-O-crotonates (Stilpnotomentolide-8-O-tiglate), 8 α-(4-hydroxyl methacryloyl)-10 Alpha-hydroxy bristle Ramulus Uncariae Cum Uncis lactone-13-acetass (α of 8 α-(4-Hydroxymethacryloyloxy)-10-hydroxyhirsutinolide-13-O-acetate), 8 α-(4-hydroxyl Fructus Crotonis acyloxy)-10 Alpha-hydroxy bristle Ramulus Uncariae Cum Uncis lactone-13-O-acetass (8 α-(4-Hydroxytigloyloxy)-10 α-hydroxyhirsutinolide-13-O-
Acetate), 8 α-(4-hydroxyl Fructus Crotonis acyloxy) bristle Ramulus Uncariae Cum Uncis lactone-13-O-acetas (8 α-(4-Hydroxytigloyloxy)-hirsutinolide-13-O-acetate), Rhizoma Cynanchi Stauntonii lactone E (GlaucolideE), 19-hydroxyl Rhizoma Cynanchi Stauntonii lactone E (19-Hydroxyglaucolide E) and Herba Vernoniae Cinereae lactone-8-O-(4-hydroxyl methacrylate) (Vernocinerolide-8-O-(4-hydroxymethacrylate)).But do not see at present and utilize chromatographic technique that this plant effective site is carried out isolating report.
Three, summary of the invention
Technical problem: the invention provides a kind of flavonoid active site that extraction obtains from ironweed Herba Vernoniae Cinereae herb, and extracting method and the application in pharmacy.
Technical scheme: technical solution of the present invention is: the flavonoid active site of a kind of Herba Vernoniae Cinereae, contain following 7 flavone compounds: the I. apiolin, quality percentage composition 1～3%, yellow needle, molecular formula is C 15H 10O 5II. golden eriodictyol, quality percentage composition 2～5%, yellow powder, molecular formula is C 16H 12O 6III. luteolin, quality percentage composition 2～5%, yellow needle, molecular formula is C 15H 10O 6IV. golden eriodictyol-7-O-β-D-glucoside, quality percentage composition 30～40%, yellow powder, molecular formula is C 22H 22O 11V. luteolin-7-O-β-D-glucoside, quality percentage composition 30～40%, yellow powder, molecular formula is C 21H 20O 11VI. Quercetin, quality percentage composition 1～3%, the khaki powder, molecular formula is C 15H 10O 7VII. apiolin-'-O-β-D-glucoside, quality percentage composition 1～3%, yellow powder, molecular formula is C 21H 20O 10
The method for preparing the flavonoid active site of above-mentioned Herba Vernoniae Cinereae, step is: it is raw material that a. takes by weighing the Herba Vernoniae Cinereae herb, dry, pulverize, with concentration expressed in percentage by volume is that 70% alcoholic solution extracts 3 times, the quality of the used alcoholic solution of each time is followed successively by 10 times, 8 times and 8 times of Herba Vernoniae Cinereae herb raw materials quality, the each extraction 2 hours merges three times extracting solution after with filtered through gauze; B. the extracting solution after the above-mentioned merging is being lower than under 80 ℃ the temperature, concentrating under reduced pressure reclaims ethanol to there not being the alcohol flavor, ethanol extraction; C. with above-mentioned ethanol extraction with distilled water diluting to density 1.10-1.20, analytical pure petroleum ether extraction 5 times, analytical pure ethyl acetate extraction 5 times, analytical pure water-saturated n-butanol with 60-90 ℃ of boiling range extracts 3 times successively, respectively petroleum ether extraction liquid, acetic acid ethyl acetate extract and water-saturated n-butanol extract are carried out concentrating under reduced pressure being lower than under 80 ℃ of temperature then, obtain petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract; D. with above-mentioned acetic acid ethyl ester extract adding distil water ultrasonic dissolution, the quality that adds distilled water is 15 times of acetic acid ethyl ester extract quality, and the centrifugal insoluble matter of removing of dissolving back tubular type obtains the ethyl acetate extraction position; E. with AB-8 macroporous adsorptive resins on the above-mentioned ethyl acetate extraction position, be 70% ethanol elution through concentration expressed in percentage by volume, obtain the flavonoid active site of Herba Vernoniae Cinereae.
The application of the flavonoid active site of above-mentioned Herba Vernoniae Cinereae in the preparation anti senile dementia drug.
Beneficial effect:
The present invention finds active good lead compound in conjunction with screening active ingredients simultaneously based on natural product research from Chinese herbal medicine, this research and development thinking is the source of original new drug exploitation.
The present invention adopts ethyl acetate extraction, macroporous adsorbent resin is refining to be separated, from ironweed Herba Vernoniae Cinereae Vernonia cinerea (L.) Less.[V.abbreviana (Wall.) DC.] prepare a kind of flavonoid active site the herb, this position mainly is made up of 8 flavone compounds, be apiolin (apigenin, I), gold eriodictyol (chrysoeriol, II), luteolin (luteolin, III), and golden eriodictyol-7-O-β-D-glucoside (thermopsoside, IV), luteolin-7-O-β-D-glucoside (luteolin-7-O-β-D-glucoside, V), and Quercetin (quercetin, VI), apiolin-'-O-β-D-glucoside (apigenin-4 '-O-β-D-glucoside, VII).Wherein compound IV obtains for separating from ironweed first.The anti-senile dementia of this active site is found active the genus first.
Five, the specific embodiment
Embodiment 1: the preparation method of flavonoid active site
Take by weighing Herba Vernoniae Cinereae herb 5.0Kg as raw material, drying, pulverize the back is 70% alcoholic solution extraction 3 times with concentration expressed in percentage by volume, the quality of three used alcoholic solution is followed successively by 10 times, 8 times and 8 times of Herba Vernoniae Cinereae herb raw materials quality, the each extraction 2 hours merges three times extracting solution after with filtered through gauze; Extracting solution after merging is being lower than under 80 ℃ the temperature, and concentrating under reduced pressure reclaims ethanol to there not being the alcohol flavor, ethanol extraction; Ethanol extraction is 1.10-1.20g/cm with distilled water diluting to density 3Analytical pure petroleum ether extraction 5 times, analytical pure ethyl acetate extraction 5 times, analytical pure water-saturated n-butanol with 60-90 ℃ of boiling range extracts 3 times successively, respectively petroleum ether extraction liquid, acetic acid ethyl acetate extract and water-saturated n-butanol extract are carried out concentrating under reduced pressure being lower than under 80 ℃ of temperature then, get petroleum ether extract 50g, acetic acid ethyl ester extract 250g, n-butyl alcohol extract 100g; The water that in acetic acid ethyl ester extract 250g, adds 15 times of quality, 50kHz, 250W ultrasonic dissolution, the centrifugal insoluble matter of removing of dissolving back tubular type gets the ethyl acetate extraction position; With AB-8 macroporous adsorbent resin on the above-mentioned ethyl acetate extraction position, resin demand is 2Kg, is 70% ethanol elution through concentration expressed in percentage by volume, obtains the flavonoid active site 50g of Herba Vernoniae Cinereae.
Embodiment 2: the structure of flavonoid active site is identified
The flavonoid active site 40g of Herba Vernoniae Cinereae, (0.063-0.200mm 250g) mixes sample with silica gel, last silicagel column (0.060-0.200mm) chromatographic isolation, (100: 0-100: 30) gradient elution, every 100mL are first-class part, wait until 105 parts altogether with methylene chloride-methanol; Methylene chloride-methanol (100: 8) part is separated with Sephadex LH-20 column chromatography through silicagel column (0.040-0.063mm), gets Compound I (15mg), II (15mg); Methylene chloride-methanol (100: 10) part is separated with the SephadexLH-20 column chromatography through silicagel column (0.040-0.063mm), gets compound III (18mg), VI (20mg); Methylene chloride-methanol (100: 20) part through silica gel (0.040-0.063mm) repeatedly column chromatography separate with Sephadex LH-20 column chromatography, get compound IV (20mg), V (15mg), VII (25mg).
Table?1? 13C-NMR(100MHz，DMSO-d 6，δppm)data?of?Flavonoids
C I II III VI IV V VII
2 163.5 163.5 163.7 147.5 162.8 164.3 164.1
3 102.7 103.1 102.7 135.5 103.7 103.1 103.0
4 5 6 7 8 9 10 1’ 2’ 3’ 4’ 5’ 6’ 181.5 160.9 98.7 163.9 93.8 157.1 103.6 121.0 128.3 115.8 161.3 115.8 128.3 181.6 161.2 98.7 163.9 93.9 157.1 103.5 121.4 110.1 150.5 147.8 115.6 120.2 181.4 161.3 98.7 164.0 93.8 157.1 103.6 121.4 113.3 145.6 149.6 115.9 118.9 175.6 155.9 98.1 163.8 93.2 160.5 102.8 121.8 114.9 144.9 145.6 115.5 119.8 181.7 160.9 99.5 163.9 94.7 156.8 105.3 122.8 112.0 151.1 146.6 113.0 118.8 181.7 156.8 99.4 162.8 94.6 160.9 105.2 119.0 113.4 145.6 149.7 115.8 121.3 181.8 156.8 99.4 162.8 94.7 160.9 105.2 120.9 128.5 115.9 161.2 115.9 128.5
1” 2” 3” 4” 5” 6” 99.8 73.0 77.1 69.5 76.3 60.6 99.8 73.0 77.0 69.5 76.3 60.5 99.8 73.0 77.1 69.5 76.4 60.5
OCH 3 55.9 55.8
Embodiment 3: the preliminary study data of the anti-senile dementia effect of flavonoid active site
Get acetic acid ethyl ester extract, macroporous resin eluting position, Compound I-VII respectively and carry out the neurocyte screening active ingredients, the result shows that all there is very strong NGF induced activity at acetic acid ethyl ester extract and macroporous resin eluting position, and Compound I-VII has certain NGF induced activity.
1, principle and method
The test of utilization neurocyte external model utilizes the PC-12 cell, detects the NGF induced activity of monomeric compound.
(1) cell strain of PC 12 cell rat adrenal medullary pheochromocytomas differentiation, it has the general features of neuroendocrine cell, and is existing to be widely used in the research of nervous physiology and neuropharmacology.
The NGF of low concentration cultivates PC 12 cells when using very, can be induced the dopaminergic neuron that is divided into NGF dependent form.
(2) LDH (Lactic dehydrogenase, lactic acid dehydrogenase) is used for the cytotoxicity of test sample.LDH content in cell culture fluid is high more, and the cytotoxicity of interpret sample is big more.
(3) mtt assay adopts this method to measure the growth inhibited effect of each sample to PC 12 cells, and DMSO does blank.
(4) receptor of Tark A NGF.The receptor of Tark B Brain-derived neurotrophic factor (BDNF is with the associated neurotrophic factor of NGF).
The fibroblast (Fibroblast) of expressing Tark A can only lean on NGF existence; The fibroblast (Fibroblast) of expressing Tark B can only lean on BDNF existence.Replace NGF or BDNF cultured cell with sample, the survival rate of cell is high more, proves that the activity of sample is high more, promptly has the NGF induced activity.
2, experimental result
Separate the biological activity assay result who obtains chemical compound in table 1 Herba Vernoniae Cinereae
Annotate: LDH is a lactic acid dehydrogenase; Trk A is 25ngmL-1 in the ED50 value that fibroblast relies on nerve growth factor NGF survival, and Trk B is 10ngmL-1 in the ED50 value that fibroblast relies on the BDNF survival; Experiment is blank with DMSO.
Embodiment 4: the preliminary pharmacodynamic study of flavonoid active site AD rat
1, materials and methods
1.1 animal: male SD rat, body weight 250 ± 20g is provided by Nanjing University of Traditional Chinese Medicine's Experimental Animal Center.
1.2 medication preparation: see embodiment 1,1.0g is equivalent to the 100g crude drug; The positive control drug duxil, by Shi Weiya (Tianjin), lot number 868159 (date of manufacture 20080730).
1.3 reagent: A β 1～40 is available from Sigma company, lot number 151-003-pc05.
1.4 grouping: 50 of SD rats are divided into normal group, model group, positive controls, treatment I at random and organize (Herba Vernoniae Cinereae active site 1.0gKg -1D -1), treatment II group (Herba Vernoniae Cinereae active site 2.0gKg -1D -1), 10 every group.
1.5 modeling method: get model group, positive controls, treatment I and organize (Herba Vernoniae Cinereae active site 1.5gKg -1D -1), treatment II group (Herba Vernoniae Cinereae active site 3.0gKg -1D -1) modeling.Concrete grammar: each is organized rat and gives 3% pentobarbital sodium 30mgKg -1Behind the intraperitoneal injection of anesthesia, with the Mus head be fixed on the stereotaxic instrument keep before and after chimney in same level, cut off scalp, expose anterior fontanelle, position by the rat brain stereotaxic atlas, at bregma rear 1.2mm, the other 2.0mm that opens of center line, bore an aperture with dental burr, vertically insert a little plastic tube, the degree of depth is 4.0mm (reaching tricorn), tubule is fixed in the rat parietal bone of head, the part is spread bactrim medicated powder and is protected from infection, and sews up scalp, and postoperative intramuscular injection every day penicillin 400,000 U/ only.Wherein, only give A β 1～40,2 μ g/ in modeling group tricorn every day, continuously 14d; Whether last injection back 7d promptly tested the 21st day, and 50 rats are carried out the ability of learning and memory test, successful to estimate modeling.
1.6 medication: begin administration after the modeling success.Treatment I group, II organize and give every day rat also fragrant cattle aqueous solution 1.5 respectively, 3.0gKg -1Irritate stomach, normal group, model group the capacity normal saline such as all give and irritate stomach; Positive controls is with duxil suspension μ gg every days 100 -1Irritate stomach, continuously 28d.
1.7 ability of learning and memory test: all rats are all in the 21st of experiment, 49d is with the length of the average escape latency in the water maze laboratory (from starting point to searching out goal platform escape required time) and enter the cecum errors number and respectively organize rat learning and memory ability assessment.Whether 21d estimates modeling successful, and 49d estimates the various influences that apply factor to ability of learning and memory.
1.8 statistical procedures: experimental data is handled through the SAS8.0 statistical package, adopts t check and q check respectively.
2, result
2.1 respectively organize the learning and memory in rats ability relatively
Table 1 is respectively organized comparison (X ± S, n=10) s of rat average escape latency
Group Dosage/gKg -1 Injection back 7d Treatment finishes
Normal group - 9.0±1.25 ** 9.3±1.25 **
Model group - 20.2±2.15 22.1±2.13
Positive controls 0.1 19.9±2.33 15.6±2.84 **
Treatment I group 2 20.0±2.05 18.1±2.23 **
Treatment II group 4 19.8±2.04 16.5±2.27 **
Annotate: *Compare with model group p＜0.01.
Table 2 is respectively organized comparison (X ± S, n=10) inferior/2min that rat enters the cecum errors number
Group Dosage/gKg -1 Injection back 7d Treatment finishes
Normal group - 3.80±1.23 ** 4.0±1.41 **
Model group - 6.40±1.71 9.3±1.49
Positive controls 0.1 6.60±1.90 6.3±2.06 **
Treatment I group 2 6.00±1.83 7.5±2.17 *
Treatment II group 4 6.70±2.00 6.9±2.60 *
Annotate: *P＜0.05, *Compare with model group p＜0.01.
2.2 conclusion
Table 1,2 shows, gives respectively organizing the rat average escape latency and entering the increase of cecum errors number of A β 1～40 injection modeling, compares with normal group, and remarkable statistical significance (p＜0.01) is arranged; After treatment finishes; treatment group rat average escape latency and enter the cecum errors number and all be starkly lower than model group; and compare with model group, have the significance statistical significance, prompting Herba Vernoniae Cinereae flavonoid active portion potential energy obviously improves the ability of learning and memory of AD rat.

Claims (3)

1. the flavone extract of a Herba Vernoniae Cinereae is characterized in that containing following 7 flavone compounds:
I. apiolin, yellow needle, molecular formula is C 15H 10O 5
II. golden eriodictyol, yellow powder, molecular formula is C 16H 12O 6
III. luteolin, yellow needle, molecular formula is C 15H 10O 6
IV. golden eriodictyol-7-O-β-D-glucoside, yellow powder, molecular formula is C 22H 22O 11
V. luteolin-7-O-β-D-glucoside, yellow powder, molecular formula is C 21H 20O 11
VI. Quercetin, the khaki powder, molecular formula is C 15H 10O 7
VII. apiolin-4 '-O-β-D-glucoside, yellow powder, molecular formula is C 21H 20O 10
Above-claimed cpd is obtained by following preparation method:
A. taking by weighing the Herba Vernoniae Cinereae herb is raw material, dry, pulverize, with concentration expressed in percentage by volume is that 70% alcoholic solution extracts 3 times, the quality of the used alcoholic solution of each time is followed successively by 10 times, 8 times and 8 times of Herba Vernoniae Cinereae herb raw materials quality, the each extraction 2 hours merges three times extracting solution after with filtered through gauze;
B. the extracting solution after the above-mentioned merging is being lower than under 80 ℃ the temperature, concentrating under reduced pressure reclaims ethanol to there not being the alcohol flavor, ethanol extraction;
C. be 1.10-1.20g/cm with above-mentioned ethanol extraction with distilled water diluting to density 3Analytical pure petroleum ether extraction 5 times, analytical pure ethyl acetate extraction 5 times, analytical pure water-saturated n-butanol with 60-90 ℃ of boiling range extracts 3 times successively, respectively petroleum ether extraction liquid, acetic acid ethyl acetate extract and water-saturated n-butanol extract are carried out concentrating under reduced pressure being lower than under 80 ℃ of temperature then, obtain petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract;
D. with above-mentioned acetic acid ethyl ester extract adding distil water ultrasonic dissolution, the quality that adds distilled water is 15 times of acetic acid ethyl ester extract quality, and the centrifugal insoluble matter of removing of dissolving back tubular type obtains the ethyl acetate extraction position;
E. with AB-8 macroporous adsorptive resins on the above-mentioned ethyl acetate extraction position, be 70% ethanol elution through concentration expressed in percentage by volume, obtain the flavone extract of Herba Vernoniae Cinereae.
2. method for preparing the flavone extract of the described Herba Vernoniae Cinereae of claim 1 is characterized in that step is:
A. taking by weighing the Herba Vernoniae Cinereae herb is raw material, dry, pulverize, with concentration expressed in percentage by volume is that 70% alcoholic solution extracts 3 times, the quality of the used alcoholic solution of each time is followed successively by 10 times, 8 times and 8 times of Herba Vernoniae Cinereae herb raw materials quality, the each extraction 2 hours merges three times extracting solution after with filtered through gauze;
B. the extracting solution after the above-mentioned merging is being lower than under 80 ℃ the temperature, concentrating under reduced pressure reclaims ethanol to there not being the alcohol flavor, ethanol extraction;
C. be 1.10-1.20g/cm with above-mentioned ethanol extraction with distilled water diluting to density 3Analytical pure petroleum ether extraction 5 times, analytical pure ethyl acetate extraction 5 times, analytical pure water-saturated n-butanol with 60-90 ℃ of boiling range extracts 3 times successively, respectively petroleum ether extraction liquid, acetic acid ethyl acetate extract and water-saturated n-butanol extract are carried out concentrating under reduced pressure being lower than under 80 ℃ of temperature then, obtain petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract;
D. with above-mentioned acetic acid ethyl ester extract adding distil water ultrasonic dissolution, the quality that adds distilled water is 15 times of acetic acid ethyl ester extract quality, and the centrifugal insoluble matter of removing of dissolving back tubular type obtains the ethyl acetate extraction position;
E. with AB-8 macroporous adsorptive resins on the above-mentioned ethyl acetate extraction position, be 70% ethanol elution through concentration expressed in percentage by volume, obtain the flavone extract of Herba Vernoniae Cinereae.
3. according to the application in the preparation anti senile dementia drug of the flavone extract of the described Herba Vernoniae Cinereae of claim 1.
CN2009100250805A 2009-02-17 2009-02-17 Flavone active site of Vernonia cinerea as well as preparation method and use thereof Expired - Fee Related CN101474196B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN2009100250805A CN101474196B (en) 2009-02-17 2009-02-17 Flavone active site of Vernonia cinerea as well as preparation method and use thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN2009100250805A CN101474196B (en) 2009-02-17 2009-02-17 Flavone active site of Vernonia cinerea as well as preparation method and use thereof

Publications (2)

Publication Number Publication Date
CN101474196A CN101474196A (en) 2009-07-08
CN101474196B true CN101474196B (en) 2010-12-01

Family

ID=40835043

Family Applications (1)

Application Number Title Priority Date Filing Date
CN2009100250805A Expired - Fee Related CN101474196B (en) 2009-02-17 2009-02-17 Flavone active site of Vernonia cinerea as well as preparation method and use thereof

Country Status (1)

Country Link
CN (1) CN101474196B (en)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104095868B (en) * 2014-07-25 2016-06-01 烟台市华文欣欣医药科技有限公司 A kind of pharmaceutical composition and pharmaceutical applications thereof improving sugar tolerance
CN105193887A (en) * 2015-09-09 2015-12-30 刘高志 Acanthopanax senticosus extract and antioxidation application thereof
CN105106276A (en) * 2015-09-23 2015-12-02 刘高志 Tradescantia extract as well as preparation method and medical application of tradescantia extract

Also Published As

Publication number Publication date
CN101474196A (en) 2009-07-08

Similar Documents

Publication Publication Date Title
CN101940625A (en) Preparation method and use of extract
CN112156156B (en) Veterinary traditional Chinese medicine antiviral oral liquid and preparation method thereof
CN104547104B (en) Ramulus Ziziphi Spinosae leaf extract and its preparation method and application
CN111110726A (en) Preparation method of lotus seed pot total flavonoids
CN101433666B (en) Anti-aging Chinese herbal medicinal composition, preparation method and use thereof
CN103040882B (en) Passiflora edulis sims stem-leaf extract as well as pharmaceutical composition and antidepressant action of extract
CN101474196B (en) Flavone active site of Vernonia cinerea as well as preparation method and use thereof
CN101480422A (en) Tibetan oriental wormwood extract as well as preparation method and use thereof
AU2002241544B2 (en) Herbal pharmaceutical compositions for treating immunological disorders
CN100441191C (en) Use of steroidal compound and total steroidal extract for preparing anti-depression drug
CN102764294B (en) Cough relieving and sputum eliminating combination and preparation method thereof
CN1293903C (en) Fant calming powder injection and its preparation method
CN102485231B (en) Discocleidion sufescens Pax et Hoffm. extract, preparation method thereof and purpose thereof
CN100335073C (en) Process for preparing semen momordicae seed extract containing triterpene saponin component
CN104804056A (en) Sarcopyramis nepalensis extract and application thereof
CN101632747B (en) Preparation technology of children cold relaxation grain
CN108619165A (en) Application of the Polysaccharide from Scutellaria Baicalensis in preparing veterinary drug
CN101928325A (en) Method for preparing natural 18-alpha glycyrrhizinic acid
CN111440186A (en) Coumarin norisoflavone compound extracted from small horse blebs and having liver protecting effect, and preparation method and application thereof
CN106822228B (en) Subprostrate sophora polysaccharide effective part and preparation method thereof
CN113288922B (en) Ephedra extract and preparation method thereof
CN104189492A (en) Traditional Chinese medicine preparation with hypoglycemic effect
CN102274254B (en) Preparation method of alligator alternanthera effective fraction extract
CN100389784C (en) Ciliate desert-grass effective parts and allergy-resistant use of effective ingredient
CN101491530A (en) Use of sterols compound and total-sterol extract in preparing analgesic

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
C17 Cessation of patent right
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20101201

Termination date: 20110217