CN1315485C - Method for preparing extracting solution of gnetum montanum which can inhibit human liver cancer cell strain - Google Patents
Method for preparing extracting solution of gnetum montanum which can inhibit human liver cancer cell strain Download PDFInfo
- Publication number
- CN1315485C CN1315485C CNB2005100279400A CN200510027940A CN1315485C CN 1315485 C CN1315485 C CN 1315485C CN B2005100279400 A CNB2005100279400 A CN B2005100279400A CN 200510027940 A CN200510027940 A CN 200510027940A CN 1315485 C CN1315485 C CN 1315485C
- Authority
- CN
- China
- Prior art keywords
- cell strain
- preparation
- hepatoma cell
- human hepatoma
- caulis gneti
- 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
Links
Landscapes
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention relates to a method for preparing gnetum montanum extract which has the effect of inhibiting liver cancer cell strains for people, which belongs to the field of techniques for preparing traditional Chinese medicines. The present invention comprises the following procedures: gnetum montanum vines are air dried, subsequently dried and then pulverized into 40 meshes of granules; mixed solvent whose weight is 10 times of that of the pulverized substances of the gnetum montanum vines is used, and the proportion by volume of ethanol to deionized water in the mixed solvent is 7:3; the mixture is heated, refluxed, extracted and concentrated in a vacuum mode, and mixed solvent extractant is concentrated into extract; ultrasonic waves are utilized, extract form substances are dissolved by the deionized water, and then, the mixture is extracted by isometric ethyl acetate; after ethyl acetate extractant is concentrated in a vacuum mode until the extractant is dry, gradient elution is carried out by taking polyamide as a constant phase and taking deionized water and ethanol as mobile phases; eluent is concentrated in a vacuum mode and freeze-dried, and sable powder substances are obtained; suspension is prepared by the RPMI1640 culture solution of fetal calf serum. The present invention has simple extraction technique, and does not need special instruments. The experiments of people tumor cell strains in vitro indicate that gnetum montanum extract whose concentration is 0.063 mg/ml has significant effect of inhibiting liver cancer cell strains BEL-7402 for people.
Description
Technical field
What the present invention relates to is the method in a kind of Chinese drug preparation technique field, specifically, is a kind of preparation method that human hepatoma cell strain is had the Caulis Gneti extractive liquid that suppresses effect.
Background technology
Caulis Gneti mainly is distributed in the Yunnan Province of China south, and province and south east asia such as Guangdong are used with Caulis Gneti Parvifolii among the people, treatment rheumatic arthritis, lumbar muscle strain, soreness of bones and muscles, traumatic injury, ulcerative hemorrhage, diseases such as bronchitis.Confirm that through the pharmacology zoopery Caulis Gneti ethanol extraction has tangible antiinflammatory action to rat Ovum Gallus domesticus album and yeast arthritis, but do not have bibliographical information at anti-tumor aspect.
Find by prior art documents, Chen Hao, Lin Mao mention in " Chinese herbal medicine " the 8th phase in 1999 " Da Ye Caulis Gneti chemical constitution study " literary composition, it is immobile phase that Da Ye Caulis Gneti crude extract is adopted polyamide, silica gel, water-ethanol, chloroform-methanol, methanol-water are that mobile phase is carried out repeatedly column chromatography, obtain 15 chemical compounds, but it is not clear and definite as yet whether to have an antitumous effect with the prepared material of this method.
Summary of the invention
The objective of the invention is to overcome the deficiencies in the prior art, a kind of preparation method that human hepatoma cell strain is had the Caulis Gneti extractive liquid that suppresses effect is provided, after making it through heating and refluxing extraction and column chromatography drip washing, improve the inhibition effect of its eluent to human hepatoma cell strain BEL-7402.
The present invention is achieved by the following technical solutions, and the present invention is a raw material with the Caulis Gneti rattan, adopts solvent extraction method, vacuum concentration, solvent extraction, lyophilization and column chromatography process to be prepared from.
Concrete steps of the present invention are as follows:
(1) rattan of Caulis Gneti is dried after, 20-30 ℃ down dry, be ground into 40 order granules again;
(2) with 10 times of mixed solvents to Caulis Gneti rattan ground product weight, the ethanol in the mixed solvent: the volume ratio of deionized water is 7: 3;
(3) 50-90 ℃ of following heating and refluxing extraction is four times, and 1 time 2 hours, 45 ℃ of following vacuum concentration were condensed into extractum with the mixed solvent extract;
(4) utilize the ultrasound wave of frequency for 100KHZ, behind deionized water dissolving cream shape thing, equal-volume ethyl acetate extraction 6 times;
(5) with behind 45 ℃ of following vacuum concentration to ten of acetic acid ethyl ester extract, polyamide (14-100 order) is an immobile phase, and deionized water-ethanol is the eluent gradient eluting;
(6) 45 ℃ of following vacuum concentration of eluent ,-40 ℃ of following lyophilizations obtain the Powdered thing of dark brown;
(7) the RPMI1640 culture fluid with 5% hyclone is made into suspension.
Find from the Caulis Gneti rattan being carried out chemical constitution study, wherein contain isorhapontigenin, resveratrol, daucosterol, stearic acid, Caulis Gneti penta element, Caulis Gneti alcohol, cupreol, Caulis Gneti third element and isorhapontigenin-3-O-β-compositions such as D-glucoside.Wherein isorhapontigenin and resveratrol etc. belong to the stilbene monomeric compound, as everyone knows, that the stilbene monomeric compound has is antibiotic, blood fat reducing, blood pressure lowering, antioxidation, free radical resisting and protein kinase inhibitory action isoreactivity, anti-inflammatory activity is in various degree still arranged in addition, particularly isorhapontigenin and resveratrol have very strong leukotriene generation inhibition and receptor acting isoreactivity, and this may be that this eluent has the reason that suppresses the active effect of human hepatoma cell strain BEL-7402.
The present invention is a raw material with the rattan of perennial wild plant Caulis Gneti, utilizes its reproducibility advantage, and abundant plant resources can be provided; Extraction process is simple, need not special instrument; In vitro tests with human tumor cell line shows: the concentration of 0.063mg/ml has significant inhibition effect to human hepatoma cell strain BEL-7402.
The specific embodiment
Embodiment 1
Take by weighing the Caulis Gneti rattan powder 200g (fine powders of 40 order granularities) of 20 ℃ of crushed after being dried, choose dehydrated alcohol: deionized water=7: 3 mixed solvent 2000ml, 50 ℃ of following heating and refluxing extraction three times, each 2 hours, 45 ℃ of following vacuum concentration reclaim solvent, get paste 80.9g, use deionized water dissolving, equal-volume ethyl acetate extraction 6 times, concentrate 10.8g extractum.Getting 1g material wherein, is leacheate with the deionization water-ethanol, and polyamide (14 order) carries out column chromatography and gets active component, concentrates, and-40 ℃ of lyophilizations get 81.4mg.
External this component 0.063mg/ml adopts sulphonyl rhodamine B protein staining method, is 87.0% to the suppression ratio of human hepatoma cell strain BEL-7402.
Embodiment 2
Take by weighing the Caulis Gneti rattan powder 200g (fine powders of 40 order granularities) of 25 ℃ of following crushed after being dried, choose dehydrated alcohol: deionized water=7: 3 mixed solvent 2000ml, 70 ℃ of following heating and refluxing extraction three times, each 2 hours, 45 ℃ of following vacuum concentration reclaim solvent, get paste 80.9g, use deionized water dissolving, equal-volume ethyl acetate extraction 6 times, concentrate 10.8g extractum.Getting 1g material wherein, is leacheate with the deionization water-ethanol, and polyamide (100 order) carries out column chromatography and gets active component, concentrates, and-40 ℃ of lyophilizations get 62.3mg.
External this component 0.25mg/ml adopts sulphonyl rhodamine B protein staining method. and the suppression ratio to human hepatoma cell strain BEL-7402 is 63.7%.
Embodiment 3
Take by weighing the Caulis Gneti rattan powder 200g (fine powders of 40 order granularities) of 30 ℃ of crushed after being dried, choose dehydrated alcohol: deionized water=7: 3 mixed solvent 2000ml, 90 ℃ of following heating and refluxing extraction three times, each 2 hours, 45 ℃ of following vacuum concentration reclaim solvent, get paste 80.9g, use deionized water dissolving, equal-volume ethyl acetate extraction 6 times, concentrate 10.8g extractum.Getting 1g material wherein, is leacheate with the deionization water-ethanol, and polyamide (40 order) carries out column chromatography and gets active component, concentrates, and-40 ℃ of lyophilizations get 71.5mg.
External this component 0.016mg/ml adopts sulphonyl rhodamine B protein staining method, is 63.1% to the suppression ratio of human hepatoma cell strain BEL-7402.
Claims (10)
1. one kind has the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that, may further comprise the steps:
(1) rattan of Caulis Gneti is dried after, drying is ground into 40 order granules again;
(2) with 10 times of mixed solvents to Caulis Gneti rattan ground product weight, mixed solvent is 70% ethanol water;
(3) heating and refluxing extraction, vacuum concentration is condensed into extractum with the mixed solvent extract;
(4) utilize ultrasound wave, behind deionized water dissolving cream shape thing, the equal-volume ethyl acetate extraction;
(5) with the acetic acid ethyl ester extract vacuum concentration after do, polyamide is an immobile phase, the deionization water-ethanol is the eluent gradient eluting;
(6) eluent vacuum concentration, lyophilization obtains the Powdered thing of dark brown;
(7) the RPMI1640 culture fluid with hyclone is made into suspension.
2. according to claim 1 have the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that, and described step (1), its baking temperature is 20-30 ℃.
3. according to claim 1 have the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that described heating and refluxing extraction, its temperature are 50-90 ℃.
4. according to claim 3 have the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that described heating and refluxing extraction is four times, 1 time 2 hours.
5. according to claim 1 have the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that, and described vacuum concentration, its temperature is 45 ℃.
6. according to claim 1 have the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that described polyamide is the 14-100 order.
7. according to claim 1 have the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that, and described ultrasound wave, its frequency is 100KHZ.
8. according to claim 1 have the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that described extraction is 6 times.
9. according to claim 1 have the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that, and described step (6), its lyophilization temperature is-40 ℃.
10. according to claim 1 have the preparation method of the Caulis Gneti extractive liquid that suppresses effect to human hepatoma cell strain, it is characterized in that, and the RPMI1640 culture fluid of described hyclone, its concentration is 5%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100279400A CN1315485C (en) | 2005-07-21 | 2005-07-21 | Method for preparing extracting solution of gnetum montanum which can inhibit human liver cancer cell strain |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100279400A CN1315485C (en) | 2005-07-21 | 2005-07-21 | Method for preparing extracting solution of gnetum montanum which can inhibit human liver cancer cell strain |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1732999A CN1732999A (en) | 2006-02-15 |
| CN1315485C true CN1315485C (en) | 2007-05-16 |
Family
ID=36075754
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005100279400A Expired - Fee Related CN1315485C (en) | 2005-07-21 | 2005-07-21 | Method for preparing extracting solution of gnetum montanum which can inhibit human liver cancer cell strain |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1315485C (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1790239B1 (en) * | 2004-09-14 | 2013-01-30 | Hosoda SHC Inc. | Gnetum extract |
| US10894067B2 (en) * | 2017-03-16 | 2021-01-19 | Yamada Bee Company, Inc. | Genome stability enhancer |
| US20210169113A1 (en) * | 2018-10-04 | 2021-06-10 | Hosoda Shc Co., Ltd. | Composition for improving intestinal flora |
| CN114052309A (en) * | 2021-11-16 | 2022-02-18 | 高梵(浙江)信息技术有限公司 | Sportswear with antibacterial and deodorant down fabric |
| CN113981703A (en) * | 2021-11-16 | 2022-01-28 | 高梵(浙江)信息技术有限公司 | Antibacterial warm-keeping jean down pants |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1566054A (en) * | 2003-06-27 | 2005-01-19 | 中国医学科学院药物研究所 | Resveratrol oligo cattail compounds, its manufacturing process, pharmaceutical combination and uses thereof |
-
2005
- 2005-07-21 CN CNB2005100279400A patent/CN1315485C/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1566054A (en) * | 2003-06-27 | 2005-01-19 | 中国医学科学院药物研究所 | Resveratrol oligo cattail compounds, its manufacturing process, pharmaceutical combination and uses thereof |
Non-Patent Citations (1)
| Title |
|---|
| 小叶买麻藤藤茎化学成分的研究 周祝等，中草药，第33卷第3期 2002 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1732999A (en) | 2006-02-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102600219B (en) | Total flavone extract of abelmoschus manihot and preparing method of total flavone extract | |
| CN1315485C (en) | Method for preparing extracting solution of gnetum montanum which can inhibit human liver cancer cell strain | |
| CN104341430A (en) | 3-phenylcoumarin robustic acid as well as extraction method and application thereof | |
| CN102119992B (en) | Pharmaceutical preparation prepared from fresh motherwort and fresh aloe and preparation method thereof | |
| CN101240005A (en) | Method for preparing platycodin D from balloon-flower root and application thereof in anti-cancer medicament | |
| Lee et al. | Butanol extracts of Asparagus cochinchinensis fermented with Weissella cibaria inhibit iNOS‑mediated COX‑2 induction pathway and inflammatory cytokines in LPS‑stimulated RAW264. 7 macrophage cells | |
| CN105131062B (en) | A kind of preparation method of Baical Skullcap root P.E | |
| CN102861123B (en) | Traditional Chinese medicine extract with effect on promoting angiogenesis as well as preparation method and application thereof | |
| CN102233013A (en) | Preparation method for total saponins of pulsatilla chinensis | |
| CN106084087A (en) | A kind of preparation method of Fructus Trichosanthis polysaccharide | |
| CN113264974B (en) | Preparation of type B cardiac glycoside and anti-angiogenesis application thereof | |
| Wu et al. | Preparation of paeoniflorin from the stems and leaves of Paeonia lactiflora Pall.‘Zhongjiang’through green efficient microwave-assisted extraction and subcritical water extraction | |
| CN101028322B (en) | Use of Maoliefengdou extract for preparing anti-cancer medicine | |
| CN101648959B (en) | Coumaronochromones compound and preparation and application thereof | |
| CN108623642A (en) | It is a kind of to extract deep congruent melting solvent aqueous mixtures of rhodioside and tyrosol and preparation method thereof and extracting method in rhodiola root for synchronous | |
| CN106110220B (en) | Preparation process of Chinese patent medicine for treating biliary tract diseases | |
| CN108836982A (en) | A kind of toad skin active component and preparation method thereof | |
| CN103479723A (en) | Diterpenoid tanshinone effective part and countercurrent chromatography preparation method and cancer treatment application thereof | |
| CN103027932A (en) | Cortex periplocae anti-tumor effective component extractive as well as preparation method and application thereof | |
| CN107556401A (en) | A kind of kuh-seng polysaccharide, its preparation method and liver protection and immunomodulation applications | |
| CN103610682A (en) | Preparation method of 3(alpha)-hydroxyl-30-olive-12,20(29)-diene-28-acid and application in preparing anti-tumor drug | |
| CN102775461A (en) | Method for preparing 20 (R)-ginseniside Rg3 | |
| CN106674305A (en) | Method for extracting monomer medicine for promoting cold acclimatization ability of organism, and application thereof | |
| CN106377523B (en) | Natural active medicine and preparation and application thereof | |
| CN101879170A (en) | Anti-tumor application of malonyl astragaloside I and aglycon thereof |
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: 20070516 Termination date: 20100721 |