CN104027477A - Camellia nitidissima tea active site, preparation method and application thereof - Google Patents
Camellia nitidissima tea active site, preparation method and application thereof Download PDFInfo
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- CN104027477A CN104027477A CN201310070480.4A CN201310070480A CN104027477A CN 104027477 A CN104027477 A CN 104027477A CN 201310070480 A CN201310070480 A CN 201310070480A CN 104027477 A CN104027477 A CN 104027477A
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Abstract
The invention discloses a Camellia nitidissima tea active site, a preparation method and application thereof. The preparation method includes: A. infiltration: taking coarse Camellia nitidissima tea powder, and soaking the powder overnight with a quintuple (V/W) alcoholic solution with a concentration of 30% or above; B. extraction: employing an alcoholic solution with a same concentration to that of step A to conduct heating reflux extraction on the coarse Camellia nitidissima tea powder in step A twice or more, with the dosage of the alcoholic solution being 10-40 times (V/W) that of the coarse Camellia nitidissima tea powder, controlling the extraction time at 1-4h each time, performing filtration, merging the extracted liquid, conducting pressure reduced concentration, recovering the solvent to obtain a crude extracted liquid, conducting filtering and taking the filtrate to obtain a liquid medicine with a proportion of 1.05-1.25; C. treatment by macroporous adsorption resin: taking the filtrate obtained in step B, performing macroporous resin column chromatography separation, carrying out elution with 10%, 30%, 80% and 95% ethanol solution with a decuple column volume in order, collecting all the elution parts respectively, and conducting pressure reduced distillation on the 80% ethanol water elution part, with the macroporous resin selected from AB-8, D101, D201, D140, D141A, D301, D401, HP20 and HPD-100; and D. drying: freeze drying the 80% ethanol part obtained in step C.
Description
Technical field
The present invention relates to active Chinese drug component position, Its Preparation Method And Use.
Background technology
A-glycosidase plays an important role in the absorption process of food, and food must be combined with this kind of enzyme the digested absorption of ability and be made blood sugar increasing.A-glycosidase inhibitor is a class new oral antidiabetic drug of current extensive use, clinical conventional have acarbose, voglibose and miglitol, its site of action is at small intestinal epimere, by reversibly suppressing the a-glycosidase of mesentery brush border, delay a-glycosidase polysaccharide (as starch, oligosaccharide etc.) is decomposed into glucose, thereby the infiltration rate of the glucose that slows down, reduces post-prandial glycemia.Acarbose mainly suppresses a-amylase, acts on the digestion process of macromolecular polysaccharide; Voglibose and miglitol optionally suppress disaccharidase hydrolytic enzyme (maltase, saccharase), and the process that makes disaccharidase be decomposed into monosaccharide is obstructed.Because this inhibitory action is reversible (a-glycosidase inhibitor dissociates again after being combined a few hours with a-glycosidase voluntarily), so carbohydrate is only to postpone to the conversion of glucose, rather than blocking-up completely.A-glycosidase inhibitor to fasting glucose without direct effect, but can reduce postprandial hyperglycemia by delaying digestion and the absorption of sugar in intestinal, alleviate the toxic action of glucose, improve insulin resistant, so the fasting glucose that makes patient also improvement to a certain extent.
Still need clinically new a-glycosidase inhibitor badly, as the medicine for the treatment of diabetes.
Guangxi is the origin center of Camellia nitidissima Chi (Camellia nitidissima chi) plant, is described as the native place of Camellia nitidissima Chi.Camellia nitidissima Chi derives from Theaceae (Theaceae) Camellia Plants; that in Flos Camelliae Japonicae family, unique petal is flavous rareness species; be listed in country-level focused protection plant, enjoy in the world " plant kingdom giant panda " and " queen's of tea family " good reputation.The successful commerial growing of Camellia nitidissima Chi at present, the functional health care product that the leaf of Camellia nitidissima Chi of take is development of raw materials, has found a good sale in all parts of the country.But not yet relevant for the research of its blood sugar lowering and inhibition a-glycosidase activity, report.
Summary of the invention
The inventor, through broad research, finds that the 80% ethanol water elution part obtaining with macroporous adsorbent resin processing again after extracting leaf of Camellia nitidissima Chi with aquiferous ethanol has good inhibitory action to a-glycosidase, thereby proposes the present invention.
The application's a embodiment provides the preparation method of Camellia nitidissima Chi active site, and it comprises the steps:
A. infiltrate: get leaf of Camellia nitidissima Chi coarse powder, by the alcoholic solution soaked overnight of 5 times of (V/W) 30% or above concentration;
B. extract: use with steps A in the alcoholic solution of same concentrations to the leaf of Camellia nitidissima Chi coarse powder heating and refluxing extraction in A step more than 2 times or 2 times, alcoholic solution consumption is 10～40 times (V/W) of leaf of Camellia nitidissima Chi coarse powder, each extraction time is 1～4 hour, filter, merge extractive liquid,, reclaims solvent, obtain crude extract, filter, get filtrate, obtaining medicinal liquid proportion is 1.05～1.25;
C. macroporous adsorbent resin is processed: by described condensed cream water suspendible, separated through macroporous resin column chromatography, use successively 10%, 30%, 80%, 95% ethanol water elution of 10 times of column volumes, collect respectively each eluate, by 80% ethanol water elution part distilling under reduced pressure; Described macroporous resin is selected from AB-8, D101, D201, D140, D141A, D301, D401, HP20 and HPD-100;
D. dry: the 80% ethanol part lyophilization that C step is obtained, obtains.
Another embodiment of the application provides the leaf of Camellia nitidissima Chi active site that adopts above-mentioned preparation method to prepare.
Another embodiment of the application provides pharmaceutical composition, the leaf of Camellia nitidissima Chi active site that it comprises the application.
Another embodiment of the application provides health product, the leaf of Camellia nitidissima Chi active site that it comprises the application.
Another embodiment of the application provides cosmetics, the leaf of Camellia nitidissima Chi active site that it comprises the application.
Another embodiment of the application provides the purposes of above-mentioned leaf of Camellia nitidissima Chi active site in the medicine for the preparation for the treatment of diabetes.This medicine can be prepared into drop pill, tablet, capsule, pill, granule, mixture or plaster.
Another embodiment of the application provides the purposes of above-mentioned leaf of Camellia nitidissima Chi active site in preparing health product.
Another embodiment of the application provides the purposes of above-mentioned leaf of Camellia nitidissima Chi active site in preparing cosmetics.
The specific embodiment
The application's a embodiment provides the preparation method of leaf of Camellia nitidissima Chi active site, and it comprises the steps:
A. infiltrate: get leaf of Camellia nitidissima Chi coarse powder, by the alcoholic solution soaked overnight of 5 times of (V/W) 30% or above concentration;
B. extract: use with steps A in the alcoholic solution of same concentrations to the leaf of Camellia nitidissima Chi coarse powder heating and refluxing extraction in A step more than 2 times or 2 times, alcoholic solution consumption is 10～40 times (V/W) of leaf of Camellia nitidissima Chi coarse powder, each extraction time is 1～4 hour, filter, merge extractive liquid,, reclaims solvent, obtain crude extract, filter, get filtrate, obtaining medicinal liquid proportion is 1.05～1.25;
C. macroporous adsorbent resin is processed: by described condensed cream water suspendible, through macroporous resin column chromatography separation, use successively 10%, 30%, 80%, 95% ethanol water elution of 10 column volumes, collect respectively each eluate, by 80% ethanol water elution part distilling under reduced pressure; Described macroporous resin is selected from AB-8, D101, D201, D140, D141A, D301, D401, HP20 and HPD-100;
D. dry: the 80% ethanol part lyophilization that C step is obtained, obtains.
In certain embodiments, described alcohol is selected from methanol, ethanol, propanol or its mixture.In a more preferred embodiment, described alcohol is ethanol.
In certain embodiments, available slow percolation or ultrasonic extraction substitute reflux, extract.
Another embodiment of the application provides the leaf of Camellia nitidissima Chi active site that adopts above-mentioned preparation method to prepare.
Another embodiment of the application provides pharmaceutical composition, the leaf of Camellia nitidissima Chi active site that it comprises the application.
Another embodiment of the application provides health product, the leaf of Camellia nitidissima Chi active site that it comprises the application.
Another embodiment of the application provides cosmetics, the leaf of Camellia nitidissima Chi active site that it comprises the application.
Another embodiment of the application provides the purposes of above-mentioned leaf of Camellia nitidissima Chi active site in the medicine for the preparation for the treatment of diabetes.This medicine can be prepared into drop pill, tablet, capsule, pill, granule, mixture or plaster.
Another embodiment of the application provides the purposes of above-mentioned leaf of Camellia nitidissima Chi active site in the medicine for the preparation of inhibition a-glycosidase.This medicine can be prepared into drop pill, tablet, capsule, pill, granule, mixture or plaster.
Another embodiment of the application provides the purposes of above-mentioned leaf of Camellia nitidissima Chi active site in preparing health product.
Another embodiment of the application provides above-mentioned leaf of Camellia nitidissima Chi active site using in preparing cosmetics to be coated with.
Embodiment
The preparation of embodiment 1 Camellia nitidissima Chi active site
Leaf of Camellia nitidissima Chi coarse powder 1000g, 5 times of 70% soak with ethanol spent the night, heating and refluxing extraction 3 times, 3h for the first time, second and third time is respectively 2.5h, and merge extractive liquid, reclaims solvent, obtain crude extract, filter, get filtrate, obtaining medicinal liquid proportion is 1.05～1.25, gained filtrate is separated through D101 macroporous resin column chromatography, use successively 10%, 30%, 80%, 95% ethanol water elution of 10 column volumes, collect respectively each eluate, by the partially recycled solvent of 80% ethanol water elution, lyophilization, obtains 80% ethanol position.
Embodiment 2 pharmacological activity tests
1) reagent
A-glycosidase (a-glucosidase, EC3.2.1.20), 4-Nitrobenzol-a-D-pyranglucoside (4-Ntrophenyl-a-D-glucopyranoside, PNPG, 026K1516), acarbose (Acarbose, Lot16869, positive control) and DMSO all purchased from Sigma company, bovine serum albumin (5 component) is (BSA) purchased from Solarbio company limited.
2) test method
By the method slightly improveing
[1]on 96 orifice plates, carry out, by the long microplate reader of all-wave (UVmax Thermo Scientific Multiskan Go), under 405nm wavelength, measure absorbance A value.
3) data analysis
According to suppression ratio %=[1-(sample A
test group-sample A
blank group)/(contrast A
test group-contrast A
blank group)] * 100% calculate suppression ratio, and with OriginPro7.5SR1 computed in software half suppression ratio concentration (IC
50) value.Meansigma methods ± standard deviation for experimental data
(n=4) represent.
Result of the test is as shown in table 1.
Table 1 leaf of Camellia nitidissima Chi active site and acarbose suppress the IC of a-glycosidase
50value
aiC
50: suppress the inhibitor concentration of a half, by pushing away and obtain in concentration-inhibition curve.All values are expressed as meansigma methods ± standard deviation (n=4).
As can be known from the above table, the activity of the application's leaf of Camellia nitidissima Chi 80% ethanol position inhibition a-glycosidase is suitable with the activity of positive control acarbose.
List of references
[1]li Ting, Zhang Xiaodong, Song Yuwen, et al.A microplate based screening method for alpha-glucosidase inhibitors (screening technique based on microwell plate of a-glycosidase inhibitor) [J] .Chinese Journal of Clinical Pharmacology and Therapeutics, 2005,10 (10): 1128-1134.
Claims (11)
1. the preparation method of leaf of Camellia nitidissima Chi active site, it comprises the steps:
A. infiltrate: get leaf of Camellia nitidissima Chi coarse powder, by the alcoholic solution soaked overnight of 5 times of (V/W) 30% or above concentration;
B. extract: use with steps A in the alcoholic solution of same concentrations to the leaf of Camellia nitidissima Chi coarse powder heating and refluxing extraction in A step more than 2 times or 2 times, alcoholic solution consumption is 10～40 times (V/W) of leaf of Camellia nitidissima Chi coarse powder, each extraction time is 1～4 hour, filter, merge extractive liquid,, reclaims solvent, obtain crude extract, filter, get filtrate, obtaining medicinal liquid proportion is 1.05～1.25;
C. macroporous adsorbent resin is processed: by described condensed cream water suspendible, separated through macroporous resin column chromatography, use successively 10%, 30%, 80%, 95% ethanol water elution of 10 times of column volumes, collect respectively each eluate, by 80% ethanol water elution part distilling under reduced pressure; Described macroporous resin is selected from AB-8, D101, D201, D140, D141A, D301, D401, HP20 and HPD-100;
D. dry: the 80% ethanol part lyophilization that C step is obtained, obtains.
2. method according to claim 1, is characterized in that described alcohol is selected from methanol, ethanol, propanol or its mixture.
3. method according to claim 1 and 2, is characterized in that, in step B, with percolation or ultrasonic extraction substitute reflux, extract, at a slow speed.
4. leaf of Camellia nitidissima Chi active site, it is according to the method preparation described in arbitrary claim in claim 1-3.
5. pharmaceutical composition, it comprises leaf of Camellia nitidissima Chi active site claimed in claim 4.
6. health product, it comprises leaf of Camellia nitidissima Chi active site claimed in claim 4.
7. cosmetics, it comprises leaf of Camellia nitidissima Chi active site claimed in claim 4.
Leaf of Camellia nitidissima Chi active site claimed in claim 4 for the preparation for the treatment of diabetes medicine in purposes.
9. purposes claimed in claim 8, wherein said medicine is prepared to drop pill, tablet, capsule, granule or mixture.
10. the purposes of leaf of Camellia nitidissima Chi active site claimed in claim 4 in preparing health product.
The purposes of 11. leaf of Camellia nitidissima Chi active sites claimed in claim 4 in preparing cosmetics.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105707973A (en) * | 2014-09-10 | 2016-06-29 | 邹玉华 | Green tea extract and preparation method thereof |
| CN108635503A (en) * | 2018-05-18 | 2018-10-12 | 大连大学 | A kind of oral honeysuckle flower tea and preparation method thereof inhibiting alpha-glucosidase |
| CN110922438A (en) * | 2019-11-22 | 2020-03-27 | 南开大学 | Method for preparing ellagic acid derivative camellia saponin from camellia chrysantha |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101558875A (en) * | 2009-05-08 | 2009-10-21 | 广西桂人堂金花茶产业集团股份有限公司 | Granule prepared by camellia chrysantha tea and preparation method thereof |
-
2013
- 2013-03-06 CN CN201310070480.4A patent/CN104027477A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101558875A (en) * | 2009-05-08 | 2009-10-21 | 广西桂人堂金花茶产业集团股份有限公司 | Granule prepared by camellia chrysantha tea and preparation method thereof |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105707973A (en) * | 2014-09-10 | 2016-06-29 | 邹玉华 | Green tea extract and preparation method thereof |
| CN108635503A (en) * | 2018-05-18 | 2018-10-12 | 大连大学 | A kind of oral honeysuckle flower tea and preparation method thereof inhibiting alpha-glucosidase |
| CN110922438A (en) * | 2019-11-22 | 2020-03-27 | 南开大学 | Method for preparing ellagic acid derivative camellia saponin from camellia chrysantha |
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