CN105348332A - Extraction method for persimmon tannin - Google Patents
Extraction method for persimmon tannin Download PDFInfo
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- CN105348332A CN105348332A CN201510759683.3A CN201510759683A CN105348332A CN 105348332 A CN105348332 A CN 105348332A CN 201510759683 A CN201510759683 A CN 201510759683A CN 105348332 A CN105348332 A CN 105348332A
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- ethanol
- persimmon
- add
- filtrate
- persimmon tannin
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H13/00—Compounds containing saccharide radicals esterified by carbonic acid or derivatives thereof, or by organic acids, e.g. phosphonic acids
- C07H13/02—Compounds containing saccharide radicals esterified by carbonic acid or derivatives thereof, or by organic acids, e.g. phosphonic acids by carboxylic acids
- C07H13/08—Compounds containing saccharide radicals esterified by carbonic acid or derivatives thereof, or by organic acids, e.g. phosphonic acids by carboxylic acids having the esterifying carboxyl radicals directly attached to carbocyclic rings
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
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- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses an extraction method for persimmon tannin. The method comprises the following steps: firstly, persimmons are peeled and crushed, ethanol is added, extraction is carried out, the extracting solution is filtered, and the filtrate is taken for standby; secondly, the filtrate is condensed, 95V% of ethanol is added, the mixture is allowed to stand overnight, centrifugation is carried out, and a supernatant is taken; thirdly, the supernatant is sieved through AB-8 macroporous resin, a column is washed by utilization of water until the column is colorless, thenthe column is washed by utilization of ethanol, and the ethanol eluate is collected; fourthly, the ethanol eluate is condensed, the solvent is recovered, an extract is obtained, the extract is dissolved in water, filtering is carried out, the filtrate is taken and is sieved through a hollow fiber film with a cut-off molecular weight of 450-600, and the cut-off solution is collected for standby; fifthly, the cut-off solution is sieved through a hollow fiber film with a cut-off molecular weight of 14000-15000, the penetration solution is collected and dried, and persimmon tannin power is prepared. The extracted persimmon tannin has a good function of inhibiting streptococcus mutans, and can be used for caries prevention.
Description
Technical field
The invention belongs to technical field of biological extraction, be specifically related to a kind of extracting method of persimmon tannin.
Background technology
The tannin of rich content in persimmon (PersimmonDiospryroskakiL) pulp.Research shows, persimmon tannin have very strong oxidation-resistance and and protein binding capacity, have suppress human lymphocyte leukemia, hypotensive, remove interior free yl and obviously suppress the activity of multiple enzyme and toxin in pallas pit viper, Naja, Agkistrodon venom.Although also report that persimmon extract, effective constituent are flavones, triterpenes and hydrolyzable tannin, show certain resistance to subtilis, resistance is also unstable.Do not see the report that persimmon tannin can be used for suppressing streptococcus mutans at present.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of extracting method of persimmon tannin, and the tannin of extraction effectively can suppress streptococcus mutans, has good prevention of dental caries effect.
Technical scheme provided by the invention is the extracting method of persimmon tannin, comprises the following steps:
1) removed the peel by persimmon, broken, add extraction using alcohol, by extracting liquid filtering, it is for subsequent use to get filtrate;
2) filtrate concentrated, add 95V% ethanol, placement is spent the night, centrifugal, gets supernatant liquor;
3) supernatant liquor is crossed AB-8 macroporous resin, first wash post with water to colourless, then use ethanol elution, collect ethanol eluate;
4) by ethanol eluate concentration and recovery solvent, obtain medicinal extract, be dissolved in water, filter, getting filtrate by molecular weight cut-off is the hollow-fibre membrane of 450 ~ 600, collects trapped fluid for subsequent use;
5) be the hollow-fibre membrane of 14000 ~ 15000 by trapped fluid by molecular weight cut-off, collect permeate, dry, be persimmon tannin powder.
Step 1) in, the volumetric concentration of ethanol is 70 ~ 80%, and its add-on is 10 ~ 20 times of persimmon weight, extracts 1 ~ 3 time, each 1 ~ 3h at 60 ~ 80 DEG C.
Step 2) in, the add-on of ethanol is 2 ~ 4 times of concentrated solution.
Step 3) in, the add-on of water is 3 ~ 5 times of medicinal extract weight.
The present invention extracts the persimmon tannin obtained and has the effect well suppressing streptococcus mutans, can be used for prevention of dental caries.
Embodiment
The present invention is further elaborated for following specific embodiment, but not as a limitation of the invention.
Embodiment 1
1) removed the peel by persimmon, broken, add the 70v% ethanol of 10 times of persimmon weight, at 60 DEG C, extract 1h, united extraction liquid, filter, it is for subsequent use to get filtrate;
2) filtrate concentrated, add 95v% ethanol, the add-on of ethanol is 2 times of concentrated solution, and placement is spent the night, centrifugal, gets supernatant liquor;
3) supernatant liquor is crossed AB-8 macroporous resin, first wash post with water to colourless, then use ethanol elution, collect ethanol eluate;
4) by ethanol eluate concentration and recovery solvent, obtain medicinal extract, be dissolved in water, the add-on of water is 3 times of medicinal extract weight, and filter, getting filtrate by molecular weight cut-off is the hollow-fibre membrane of 450 ~ 600, collects trapped fluid for subsequent use;
5) be the hollow-fibre membrane of 14000 ~ 15000 by trapped fluid by molecular weight cut-off, collect permeate, vacuum lyophilization, is persimmon tannin powder.
Embodiment 2
1) removed the peel by persimmon, broken, add the 80v% ethanol of 20 times of persimmon weight, extract 3 times, each 3h, united extraction liquid at 80 DEG C, filter, it is for subsequent use to get filtrate;
2) filtrate concentrated, add 95v% ethanol, the add-on of ethanol is 4 times of concentrated solution, and placement is spent the night, centrifugal, gets supernatant liquor;
3) supernatant liquor is crossed AB-8 macroporous resin, first wash post with water to colourless, then use ethanol elution, collect ethanol eluate;
4) by ethanol eluate concentration and recovery solvent, obtain medicinal extract, be dissolved in water, the add-on of water is 5 times of medicinal extract weight, and filter, getting filtrate by molecular weight cut-off is the hollow-fibre membrane of 450 ~ 600, collects trapped fluid for subsequent use;
5) be the hollow-fibre membrane of 14000 ~ 15000 by trapped fluid by molecular weight cut-off, collect permeate, vacuum lyophilization, is persimmon tannin powder.
Embodiment 3
1) removed the peel by persimmon, broken, add the 75v% ethanol of 15 times of persimmon weight, extract 2 times, each 2h, united extraction liquid at 70 DEG C, filter, it is for subsequent use to get filtrate;
2) filtrate concentrated, add 95v% ethanol, the add-on of ethanol is 3 times of concentrated solution, and placement is spent the night, centrifugal, gets supernatant liquor;
3) supernatant liquor is crossed AB-8 macroporous resin, first wash post with water to colourless, then use ethanol elution, collect ethanol eluate;
4) by ethanol eluate concentration and recovery solvent, obtain medicinal extract, be dissolved in water, the add-on of water is 4 times of medicinal extract weight, and filter, getting filtrate by molecular weight cut-off is the hollow-fibre membrane of 450 ~ 600, collects trapped fluid for subsequent use;
5) be the hollow-fibre membrane of 14000 ~ 15000 by trapped fluid by molecular weight cut-off, collect permeate, vacuum lyophilization, is persimmon tannin powder.
Comparative examples 1
1) removed the peel by persimmon, broken, add the 75v% ethanol of 15 times of persimmon weight, extract 2 times, each 2h, united extraction liquid at 70 DEG C, filter, it is for subsequent use to get filtrate;
2) filtrate concentrated, add 95v% ethanol, the add-on of ethanol is 3 times of concentrated solution, and placement is spent the night, centrifugal, gets supernatant liquor;
3) supernatant liquor is crossed AB-8 macroporous resin, first wash post with water to colourless, then use ethanol elution, collect ethanol eluate;
4) by ethanol eluate concentration and recovery solvent, vacuum lyophilization, is persimmon tannin powder.
Experimental example (extracorporeal bacteria inhibitor test)
1, experimental technique:
1) the persimmon tannin powder 1g of Example 1 ~ 3 and comparative examples 1 adds 10ml sterilized water and is made into mixing solutions after pulverizing respectively.
2) preparation of bacteria inhibition tablet: get the aseptic and filter paper 4 of drying, every sheet drips the mixing solutions 20 μ l that embodiment 1 ~ 3 and comparative examples 1 are made respectively, then filter paper is lain against in clean aseptic plane ware, uncap to put in incubator (37 DEG C) and dry, or put under room temperature for subsequent use after seasoning.The preparation of negative control group bacteria inhibition tablet: get aseptic dry filter paper sheet, every sheet drips sterile distilled water 20 μ l, for subsequent use after dry.
3) streptococcus mutans is inoculated: dipping concentration with sterile cotton swab is 5 × 10
5~ 5 × 10
6cfu/ml streptococcus mutans bacteria suspension, nutrient agar planar surface uniform application 3 times, often smears once, dull and stereotypedly rotates 60 °, finally cotton swab is smeared one week around plate edge, builds plate, put drying at room temperature 5min.
4) bacteria inhibition tablet is placed with: be placed with a microbiological contamination flat board at every turn, and each flat board is placed with 4 bacteria inhibition tablets and 1 negative control group bacteria inhibition tablet of embodiment and comparative examples.Be placed with in planar surface with aseptic nipper coupongs, at a distance of more than 25mm between each print center, with periphery more than the 15mm apart of flat board.After being placed with, with the light pressure-like sheet of aseptic nipper, making it be close to planar surface, build plate, be placed in 37 DEG C of incubators and cultivate 16 ~ 18h, with the diameter (comprising paster) of vernier caliper measurement bacterial restrain and record, experiment in triplicate.
2, experimental result
| Inhibition zone (mm) | |
| Embodiment 1 | 15.8 |
| Embodiment 2 | 16.0 |
| Embodiment 3 | 16.1 |
| Comparative examples 1 | 6.7 |
| Negative control group | 3.8 |
Claims (4)
1. the extracting method of persimmon tannin, is characterized in that: comprise the following steps:
1) removed the peel by persimmon, broken, add extraction using alcohol, by extracting liquid filtering, it is for subsequent use to get filtrate;
2) filtrate concentrated, add the ethanol of 95v%, placement is spent the night, centrifugal, gets supernatant liquor;
3) supernatant liquor is crossed AB-8 macroporous resin, first wash post with water to colourless, then use ethanol elution, collect ethanol eluate;
4) by ethanol eluate concentration and recovery solvent, obtain medicinal extract, be dissolved in water, filter, getting filtrate by molecular weight cut-off is the hollow-fibre membrane of 450 ~ 600, collects trapped fluid for subsequent use;
5) be the hollow-fibre membrane of 14000 ~ 15000 by trapped fluid by molecular weight cut-off, collect permeate, dry, be persimmon tannin powder.
2. the extracting method of persimmon tannin according to claim 1, is characterized in that: step 1) in, the volumetric concentration of ethanol is 70 ~ 80%, and its add-on is 10 ~ 20 times of persimmon weight, extracts 1 ~ 3 time, each 1 ~ 3h at 60 ~ 80 DEG C.
3. the extracting method of persimmon tannin according to claim 1, is characterized in that: step 2) in, the add-on of ethanol is 2 ~ 4 times of concentrated solution.
4. the extracting method of persimmon tannin according to claim 1, is characterized in that: step 3) in, the add-on of water is 3 ~ 5 times of medicinal extract weight.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510759683.3A CN105348332A (en) | 2015-11-10 | 2015-11-10 | Extraction method for persimmon tannin |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510759683.3A CN105348332A (en) | 2015-11-10 | 2015-11-10 | Extraction method for persimmon tannin |
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| CN105348332A true CN105348332A (en) | 2016-02-24 |
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| CN201510759683.3A Pending CN105348332A (en) | 2015-11-10 | 2015-11-10 | Extraction method for persimmon tannin |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106008616A (en) * | 2016-07-05 | 2016-10-12 | 深圳市美漾无华生物科技有限公司 | Method for extracting persimmon tannin from persimmons |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20110123447A (en) * | 2010-05-07 | 2011-11-15 | 왕성우 | Tannin extracting method from the wasted peels of persimmon |
| CN104478950A (en) * | 2014-12-19 | 2015-04-01 | 桂林得坤生物科技股份有限公司 | Method for extracting persimmon tannin high in small molecular tannin content from persimmon |
| CN104788511A (en) * | 2015-03-31 | 2015-07-22 | 桂林得坤生物科技股份有限公司 | Method for preparing persimmon tannins with high micromolecular tannin content from persimmons |
-
2015
- 2015-11-10 CN CN201510759683.3A patent/CN105348332A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20110123447A (en) * | 2010-05-07 | 2011-11-15 | 왕성우 | Tannin extracting method from the wasted peels of persimmon |
| CN104478950A (en) * | 2014-12-19 | 2015-04-01 | 桂林得坤生物科技股份有限公司 | Method for extracting persimmon tannin high in small molecular tannin content from persimmon |
| CN104788511A (en) * | 2015-03-31 | 2015-07-22 | 桂林得坤生物科技股份有限公司 | Method for preparing persimmon tannins with high micromolecular tannin content from persimmons |
Non-Patent Citations (1)
| Title |
|---|
| 周湘: "柿果实不同部分鞣质含量的测定及其水提取物对口腔致病菌体外抑菌活性的研究", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106008616A (en) * | 2016-07-05 | 2016-10-12 | 深圳市美漾无华生物科技有限公司 | Method for extracting persimmon tannin from persimmons |
| CN106008616B (en) * | 2016-07-05 | 2018-08-07 | 深圳市美漾无华生物科技有限公司 | A method of extracting persimmon tannin from persimmon |
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Application publication date: 20160224 |
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