CN1089006C - Method of extracting activated hemagglutination factor and inhibitor component (ATI) from tea - Google Patents
Method of extracting activated hemagglutination factor and inhibitor component (ATI) from tea Download PDFInfo
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- CN1089006C CN1089006C CN99111391A CN99111391A CN1089006C CN 1089006 C CN1089006 C CN 1089006C CN 99111391 A CN99111391 A CN 99111391A CN 99111391 A CN99111391 A CN 99111391A CN 1089006 C CN1089006 C CN 1089006C
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- folium camelliae
- camelliae sinensis
- chloroform
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Abstract
The present invention relates to a method of extracting activated hemagglutination factors and inhibitor components (ATI) from tea, which belongs to the technical field of biology. Fxa inhibitor components with obvious selectivity are obtained by three steps of reagent extraction conversion of absolute alcohol/chloroform/water. The present invention has the characteristics of simple and convenient step of separation and purification, and specific detection, and has the potential of developing new medicine.
Description
This method is about extracting the problem of activatory thrombin ten constituents for suppressing, belonging to biological technical field.
Along with development of science and technology, natural activatory thrombin ten (FXa) inhibitor becomes the recent studies on object of anticoagulation medicine.
Document was once arranged
[1,2]Report extracts a kind of substance A ntistasin and have the effect that suppresses FXa from the saliva of Mexico Hirudo, but this material content in the saliva of Hirudo is lower, and leaching process is loaded down with trivial details simultaneously, spends higher.Owing to cysteine residues content in its component is very high, the cost of clonal expression is very high again, and it is very limited when using.So we need a kind of quick, easy, economic purification to prepare the method for FXa constituents for suppressing.
Folium Camelliae sinensis is drunk by Chinese people very early as a kind of beverage.Pharmacological action that it is generally acknowledged Folium Camelliae sinensis mainly contains excitor nerve central action blood vessel dilating, effects such as diuresis and detoxifcation
[3]We find to contain in the Folium Camelliae sinensis FXa constituents for suppressing.
The present invention be a kind of from the green tea of Anhui the method for activatory thrombin ten constituents for suppressing of purifies and separates.Easy acquisition of its raw material and price are relatively low, transform by the reagent extraction of three step of dehydrated alcohol/chloroform/water, can obtain obviously optionally FXa constituents for suppressing.The step of separation and purification is easy, detects clear and definite.The activated clotting factor constituents for suppressing that extracts from Folium Camelliae sinensis is the very high inhibitor of a kind of selectivity.Fig. 1 is the activity inhibition curve of ATI to activatory thrombin ten, and Fig. 2 is the inhibition curve of ATI (extraction sample) to thrombin activity, and Fig. 3 is Ca
2+ATI suppresses curve to the activity of activatory thrombin ten when excessive.Even the detection that it is suppressed level has good restraining activated clotting factor ten active effects, as can be known at Ca
2+Also be like this under the excessive greatly situation.Its maximum characteristics are only FXa to be had very strong inhibitory action, and are then lower to the inhibitory action of thrombin.Activatory thrombin ten inhibitor that exist in blood as antithrombin-III, but do not show this species specificity.FXa is on the joint of inside and outside source blood coagulation path, does not intersect and do not have with fibrinolytic system, so it has very strong uniqueness., also might be a kind of good medicine guide component just because it is difficult for being consumed by other enzyme.
Do not find from Folium Camelliae sinensis, to extract the report of FXa constituents for suppressing at present both at home and abroad as yet, the experiment proved that and do not contain albumen in the component that illustrate that it is non-protein compound, this also is to find first in the world.
Purpose of the present invention will obtain highly active activated clotting factor ten constituents for suppressing by a simple separating step exactly.Because its specificity, high efficiency and economic dispatch characteristics probably can become an interesting break-through point in the new drug development.Enforcement schemes of the present invention:
With Anhui commercially available Hemerocallis citrina Baroni cloud point green tea fragmentation, 1. with 400 milliliters of immersions of the every hectogram Folium Camelliae sinensis of dehydrated alcohol.Soak and at room temperature carry out, after 4 hours, fully stirred 30 minutes, filter.2. every then hectogram Folium Camelliae sinensis extracts once more with 100 milliliters of dehydrated alcohol, after 1 hour, stirs filtration in 15 minutes.2. step carry out twice, obtain dehydrated alcohol tea extraction liquid.
The distilling under reduced pressure under 50 ℃ of conditions of dehydrated alcohol tea extraction liquid is concentrated into about 100 milliliters, adds 300 milliliters of chloroforms, fully shake up, and be placed in 500 milliliters of separatory funnels.100 milliliters of redistilled waters are joined chloroform/dehydrated alcohol Folium Camelliae sinensis concentrate in the extraction solution, fully shake to solution and present complete emulsified state, standing demix can be discharged chloroform layer from the bottom of separatory funnel, stay to be flaxen aqueous layer.
The aqueous composition lyophilization is obtained about 2.2 gram solid contents, and sealing is kept under-20 ℃.Sample after the lyophilization suppresses the detection of level, sees accompanying drawing 1, accompanying drawing 2 and accompanying drawing 3.From accompanying drawing 1,2,3 as can be seen, the sample after the lyophilization has the level of the activatory thrombin ten of fabulous inhibition, even at Ca
2+Under the excessive greatly situation, also be like this.Sample then can not show a candle to activatory thrombin ten inhibition of thrombin activity.
List of references: [1] Tuszynski P.G.Gasic, B.T.And Gasic, J.G.J.Biol.Chem., 262 (20), 9718 (1987) [2] Kaiser B.Seminars in Thrombosis and Hemostasis, 17 (2), the new medical college in 130 (1991) [3] Jiangsu is compiled, the Chinese medicine voluminous dictionary, Shanghai: Science and Technology of Shanghai publishing house publishes (1977), p1601
Claims (2)
1. a method of extracting activatory thrombin ten constituents for suppressing from Folium Camelliae sinensis is extracted Folium Camelliae sinensis with soaked in absolute ethyl alcohol, after filtering and concentrating, adds chloroform, fully shakes, and standing demix is discharged chloroform, collects remaining aqueous layer, and lyophilization promptly.
2. according to claim 1, the employed raw material of this method is a Chinese green tea.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN99111391A CN1089006C (en) | 1999-08-13 | 1999-08-13 | Method of extracting activated hemagglutination factor and inhibitor component (ATI) from tea |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN99111391A CN1089006C (en) | 1999-08-13 | 1999-08-13 | Method of extracting activated hemagglutination factor and inhibitor component (ATI) from tea |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1284368A CN1284368A (en) | 2001-02-21 |
| CN1089006C true CN1089006C (en) | 2002-08-14 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN99111391A Expired - Fee Related CN1089006C (en) | 1999-08-13 | 1999-08-13 | Method of extracting activated hemagglutination factor and inhibitor component (ATI) from tea |
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| CN (1) | CN1089006C (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1038649A (en) * | 1988-06-16 | 1990-01-10 | 默里尔多药物公司 | By extracting and the purifying anticoagulant composition in the leech of South America |
| CN1178467A (en) * | 1995-03-14 | 1998-04-08 | 因迪纳有限公司 | Polyphenol fractions of tea, the use thereof and formulations containing same |
| DE29819590U1 (en) * | 1998-11-03 | 1999-02-04 | Stratmann Rembert Dr | Green tea concentrate |
-
1999
- 1999-08-13 CN CN99111391A patent/CN1089006C/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1038649A (en) * | 1988-06-16 | 1990-01-10 | 默里尔多药物公司 | By extracting and the purifying anticoagulant composition in the leech of South America |
| CN1178467A (en) * | 1995-03-14 | 1998-04-08 | 因迪纳有限公司 | Polyphenol fractions of tea, the use thereof and formulations containing same |
| DE29819590U1 (en) * | 1998-11-03 | 1999-02-04 | Stratmann Rembert Dr | Green tea concentrate |
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| Publication number | Publication date |
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| CN1284368A (en) | 2001-02-21 |
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