CN104860916A - Method for extracting proanthocyanidins from apple peel - Google Patents
Method for extracting proanthocyanidins from apple peel Download PDFInfo
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- CN104860916A CN104860916A CN201510160139.7A CN201510160139A CN104860916A CN 104860916 A CN104860916 A CN 104860916A CN 201510160139 A CN201510160139 A CN 201510160139A CN 104860916 A CN104860916 A CN 104860916A
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- pycnogenols
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
- C07D311/60—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2
- C07D311/62—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2 with oxygen atoms directly attached in position 3, e.g. anthocyanidins
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Abstract
The invention relates to a method for extracting proanthocyanidins from a plant, specifically to a method for extracting proanthocyanidins from apple peel, belonging to the field of biochemical industry. The invention creatively discloses the method for extracting proanthocyanidins in the apple peel by using a microwave-enzyme combined process. According to the method, after enzymolysis of the apple peel with compound enzyme, proanthocyanidins in the apple peel is extracted through microwave-assisted extraction, and the compound enzyme is composed of cellulase, hemicellulase and pectase. The method provided by the invention fills a technical gap in extraction of proanthocyanidins from the apple peel, provides important technical guidance to extraction of proanthocyanidins from fruit peel, realizes high-yield and damage-free extraction of proanthocyanidins from the apple peel, and has a significant economic value.
Description
Technical field
The present invention relates to the method extracting pycnogenols from plant, be specifically related to a kind ofly bring up again the method for getting pycnogenols from Pericarpium Mali pumilae, belong to biological chemical field.
Background technology
Pycnogenols (Oligomer Procyanidoic Complexes), being called for short OPC, is a kind of Vitamin P complex having special molecular structure, is the most effective natural antioxidants of removing people interior free yl of generally acknowledging in the world at present.Be generally red-brown powder, gas be micro-, puckery, water-soluble and most organic solvent.
Pycnogenols is applied very extensive in life, and such as, in skin, pycnogenols has unique chemistry and physiologically active, in skin care product, play multiple action, as anti-ageing, uvioresistant, radioprotective, brighten, moisturizing etc.Pycnogenols has unique effect to the skin aging that many factors causes; Pycnogenols has propagation and refresh function to hair follicle cell, thus can promote hair growth and regeneration etc.So be highly significant and prospect for the research of pycnogenols.
Pycnogenols all has distribution in the plants such as Semen Vitis viniferae, what at present research was more is how separation and Extraction pycnogenols from Semen Vitis viniferae.
Along with the expansion of plant research scope, also there is pycnogenols in increasing fundamental research display Pericarpium Mali pumilae at present.Apple is the maximum fruit of China's output, and resource is very abundant, has good nutritive value and special health-care effect, and the polyphenol in apple can the propagation of anticancer.Wherein, pycnogenols is the major phenolic composition in apple, has multiple biological activity, has very strong antioxidant effect.Because apple young fruit is rich in pycnogenols, after ripe apples, pycnogenols is mainly present in pericarp.Therefore, extract pycnogenols with Pericarpium Mali pumilae and there is far-reaching economic benefit and social benefit.
But, for the research of Pericarpium Mali pumilae procyanidins or blank in prior art, and pycnogenols extraction process comparatively ripe is at present with this kind of seed of Semen Vitis viniferae for Research foundation, therefore more weak to the guide for method reference significance extracting pycnogenols from Pericarpium Mali pumilae.
Summary of the invention
The object of the invention is to fill up the blank in current technology, creationary a kind of method extracting pycnogenols from Pericarpium Mali pumilae is disclosed, described extracting method is to extract the pycnogenols in Pericarpium Mali pumilae with microwave combined enzyme process, described extracting method be by Pericarpium Mali pumilae after the enzymolysis of prozyme, pycnogenols wherein, the prozyme that described enzyme is made up of cellulase, hemicellulase and polygalacturonase is extracted again in the mode of microwave extracting.
As the preferred mode of one, we disclose described enzyme by etc. the prozyme that forms of the cellulase of quality, hemicellulase and polygalacturonase.
Meanwhile, the optimum condition that we also disclosed described enzymolysis is pH 4.5 ~ 5.0, enzymolysis time 40 ~ 50 minutes, hydrolysis temperature 40 ~ 60 DEG C.
Further, the most preferably condition is pH 5.0, enzymolysis time 50 minutes, hydrolysis temperature 50 DEG C.
In addition, we also the addition of preferred described prozyme be add prozyme 3mg in every 100g apple.Wherein particularly preferred mode is that described prozyme is made up of cellulase 1mg, hemicellulase 1mg, polygalacturonase 1mg.
In addition, in microwave extracting, we are also preferred, and described microwave power is 300W.
Finally, we disclose the concrete technology that this microwave combined enzyme process extracts pycnogenols from Pericarpium Mali pumilae, comprise the following steps:
(1) by Pericarpium Mali pumilae pulverize, make powder after sieve, add the buffered soln that pH is 4.5 ~ 5.0;
(2) in the buffered soln of Pericarpium Mali pumilae, the complex enzyme liquid of cellulase, hemicellulose, polygalacturonase formation is added, under the condition of 40 ~ 60 DEG C, enzymolysis 40 ~ 50 minutes;
(3) by the mixed solution after enzymolysis at microwave power 300W, frequency 2450MHz, under the condition that temperature is 60 ~ 80 DEG C extract 2 ~ 5 minutes;
(4) by extraction liquid filtered while hot, and filtrate is centrifugal under the condition of 3000r/min, get supernatant liquid, be pycnogenols solution.
Preferably, extraction time is 3 minutes; The add-on of buffered soln and the solid-liquid ratio of Pericarpium Mali pumilae powder are 1g/10mL.
Adopting the method disclosed in the present, filled up the technological gap extracting pycnogenols at present from Pericarpium Mali pumilae, also providing important technology guide for extracting pycnogenols from fruit skin simultaneously.Utilize this method can high yield, without destroy from Pericarpium Mali pumilae extract obtain pycnogenols, there is important economic worth.
Embodiment
Material used in embodiment and reagent, instrument source.
Apple, purchased from Nanjing Xiaozhuang College's fruit supermarket, obtains Pericarpium Mali pumilae by oneself, pulverizes, and crosses 40 ~ 60 mesh sieves, as Pericarpium Mali pumilae sample to be tested.
Reagent: dehydrated alcohol, Nanjing Chemistry Reagent Co., Ltd.;
Methyl alcohol, Nanjing Chemistry Reagent Co., Ltd.;
Propyl carbinol, Nanjing Chemistry Reagent Co., Ltd.;
Hydrochloric acid, pilot scale chemical corp, Shanghai;
Ferric ammonium sulfate, Chemical Reagent Co., Ltd., Sinopharm Group;
Pycnogenols standard specimen, Nat'l Pharmaceutical & Biological Products Control Institute;
Above reagent is analytical pure.
Cellulase, enzyme work >=30000u/g; Polygalacturonase, enzyme work >=30000u/g; Hemicellulase, enzyme work >=10000u/g; Proteolytic enzyme, enzyme work >=50000u/g, upper sea blue season development in science and technology company limited.
Instrument: electronic balance, Japanese Shimadzu Corporation;
7230G visible spectrophotometer, Shanghai exact science company limited;
China of state digital display thermostat water bath HH-4, Guo Hua Electrical Appliances Co., Ltd;
Desk centrifuge, Town in Shanghai booth tech equipment factory; Zhengzhou Greatwall Scientific Industrial & Trading Co., Ltd.;
Microwave synthesize abstraction instrument, Xinyi Microwave Chemistry Tech Co., Ltd..
Embodiment 1
Take pycnogenols standard specimen 1.0000mg, be dissolved in 70% ethanolic soln, be settled to 50mL, obtain pycnogenols standard reserving solution, the concentration of its procyanidins is 0.02mg/ml.
Accurately take pycnogenols standard specimen 5mg, be settled to 25mL, obtained concentration is the reference liquid of 0.2mg/ml, draw the reference liquid 0,0.2ml, 0.4ml of 0.2mg/ml respectively, 0.6ml, 0.8ml, 1.0ml, 1.2ml, and be settled to 10ml with the ethanolic soln dilution of 70%, making concentration is 0,0.004,0.008,0.012, the reference liquid of 0.016,0.02,0.024mg/ml.
The reference liquid of different concns is drawn 0.2ml respectively, be placed in the colorimetric cylinder of 7 10ml tool plugs respectively, respectively add 1.0mL methanol solution, add the n-butyl alcohol-hydrochloric acid solution (95/5 of 6.0ml again, and 2% ammonium ferric sulfate solution of 0.2ml V/V), after shaking up, put boiling water bath 95 DEG C heating 40min, then cool rapidly, standard solution is replaced to make blank with methyl alcohol, absorbancy is measured in maximum absorption wavelength 550nm place, drawing standard curve also calculates, obtaining regression equation is A(absorbancy)=3.1705C(mg/ml)+0.0023, coefficient R 2=0.9992.
Embodiment 2
Pericarpium Mali pumilae is pulverized, sieved, and accurately take 100g, the pH adding 10ml is the buffered soln of 5.0,50 DEG C of water-bath concussions, adds the complex enzyme liquid of 1mg cellulase, 1mg hemicellulose, the formation of 1mg polygalacturonase, under the condition of 50 DEG C, and enzymolysis 40 ~ 50 minutes;
By enzymolysis solution at microwave power 200 ~ 400W, frequency 2450MHz, temperature 60 ~ 80 DEG C, condition under extraction 2 ~ 5 minutes;
By extraction liquid filtered while hot, and filtrate is centrifugal under the condition of 3000r/min, get supernatant liquid, obtain pycnogenols solution.
Gained pycnogenols solution is measured absorbancy in maximum absorption wavelength 550nm place.And utilize regression equation for A(absorbancy)=3.1705C(mg/ml)+0.0023, the content calculating pycnogenols is 12.6mg.
Therefore, adopt in the present embodiment the pycnogenols that obtains yield be 12.6mg/100g.
Embodiment 3
Operate according to the mode in embodiment 2.
As different from Example 2, respectively with different cellulases, hemicellulase, polygalacturonase composition prozyme.
Table 1: different complex enzyme formula lists
| Test | A cellulase enzyme concentration mg | B hemicellulase enzyme concentration mg | C polygalacturonase enzyme concentration mg | Yield mg/100g |
| Simultaneous test 1 | 1 | 1 | 1 | 12.6 |
| Simultaneous test 2 | 1 | 2 | 2 | 9.60 |
| Simultaneous test 3 | 1 | 3 | 3 | 8.90 |
| Simultaneous test 4 | 2 | 1 | 2 | 11.0 |
| Simultaneous test 5 | 2 | 2 | 3 | 10.8 |
| Simultaneous test 6 | 2 | 3 | 1 | 11.3 |
| Simultaneous test 7 | 3 | 1 | 3 | 9.80 |
| Simultaneous test 8 | 3 | 2 | 1 | 10.3 |
| Simultaneous test 9 | 3 | 3 | 2 | 10.6 |
By the data provided in above table, can find out and adopt technology yield disclosed in this invention all at more than 8.9mg/100g, meet industrialization demand, there is very high industrialized popularization value.We also find simultaneously, adopt the cellulase of equivalent, hemicellulase, its yield of polygalacturonase far above other proportioning situations, have had the raising of significance, can reach 12.6mg/100g.
Embodiment 4
Operate according to the mode in embodiment 2.
As different from Example 2, different enzymatic hydrolysis conditions is adopted respectively.
Table 2: different enzymatic hydrolysis condition lists
| Test | Enzymolysis time (min) | Enzymolysis pH | Hydrolysis temperature (DEG C) | Content mg/100g |
| Simultaneous test 11 | 40 | 4.5 | 40 | 11.3 |
| Simultaneous test 12 | 50 | 5 | 50 | 12.6 |
| Simultaneous test 13 | 60 | 5.5 | 60 | 10.9 |
| Simultaneous test 14 | 40 | 5 | 60 | 9.50 |
| Simultaneous test 15 | 50 | 5.5 | 40 | 9.80 |
| Simultaneous test 16 | 60 | 4.5 | 50 | 10.2 |
| Simultaneous test 17 | 40 | 5.5 | 50 | 8.80 |
| Simultaneous test 18 | 50 | 4.5 | 60 | 9.30 |
| Simultaneous test 19 | 60 | 5 | 40 | 8.40 |
Can be found out by above-mentioned simultaneous test data, when employing hydrolysis temperature 50 DEG C, pH is 5, and after the enzymolysis enzymatic hydrolysis condition of 50 minutes, yield can reach 12.6mg/100g, has significant technique effect.Also can obtain the data of yield at 8.4mg/100g under other conditions, meet the needs of industrialization promotion.
Claims (10)
1. one kind is extracted the method for pycnogenols from Pericarpium Mali pumilae, it is characterized in that, described extracting method be by Pericarpium Mali pumilae after the enzymolysis of prozyme, pycnogenols wherein, the prozyme that described enzyme is made up of cellulase, hemicellulase and polygalacturonase is extracted again in the mode of microwave extracting.
2. method according to claim 1, is characterized in that, described enzyme by etc. the prozyme that forms of the cellulase of quality, hemicellulase and polygalacturonase.
3. method according to claim 1 and 2, is characterized in that, the condition of described enzymolysis is: pH condition 4.5 ~ 5.0, enzymolysis time 40 ~ 50 minutes, hydrolysis temperature 40 ~ 60 DEG C.
4. method according to claim 3, is characterized in that, described enzymatic hydrolysis condition is: pH condition 5.0, enzymolysis time 50 minutes, hydrolysis temperature 50 DEG C.
5. method according to claim 1, is characterized in that, the addition of described prozyme is add prozyme 3mg in every 100g apple.
6. method according to claim 5, is characterized in that, described prozyme is made up of cellulase 1mg, hemicellulase 1mg, polygalacturonase 1mg.
7. method according to claim 1, is characterized in that, in described microwave extracting, microwave power is 300W.
8. method according to claim 1, is characterized in that, comprises the following steps:
(1) by Pericarpium Mali pumilae pulverize, make powder after sieve, add the buffered soln that pH is 4.5 ~ 5.0;
(2) in the buffered soln of Pericarpium Mali pumilae, the complex enzyme liquid of cellulase, hemicellulose, polygalacturonase formation is added, under the condition of 40 ~ 60 DEG C, enzymolysis 40 ~ 50 minutes;
(3) by the mixed solution after enzymolysis at microwave power 300W, frequency 2450MHz, under the condition that temperature is 60 ~ 80 DEG C extract 2 ~ 5 minutes;
(4) by extraction liquid filtered while hot, and filtrate is centrifugal under the condition of 3000r/min, get supernatant liquid, be pycnogenols solution.
9. method according to claim 8, is characterized in that, extraction time is 3 minutes.
10. method according to claim 8, is characterized in that, the add-on of buffered soln and the solid-liquid ratio of Pericarpium Mali pumilae powder are 1g/10mL.
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| CN201510160139.7A CN104860916A (en) | 2015-04-07 | 2015-04-07 | Method for extracting proanthocyanidins from apple peel |
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| CN201510160139.7A CN104860916A (en) | 2015-04-07 | 2015-04-07 | Method for extracting proanthocyanidins from apple peel |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105510319A (en) * | 2015-12-07 | 2016-04-20 | 湖南科技大学 | Method for quickly identifying color of seed coats of rape seeds in early development period |
| CN106699719A (en) * | 2016-11-29 | 2017-05-24 | 蓬莱海洋（山东）股份有限公司 | Method for efficiently extracting procyanidine ingredient from young apple fruits removed by thinning |
| CN106749151A (en) * | 2016-11-28 | 2017-05-31 | 沈阳农业大学 | A kind of rapid extracting method of blueberry pomace anthocyanidin |
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| CN1395573A (en) * | 2000-01-11 | 2003-02-05 | 比奥雷克斯健康有限公司 | Extraction of flavonoids |
| CN104109403A (en) * | 2013-12-10 | 2014-10-22 | 大兴安岭神州北极蓝莓生物工程有限公司 | New wild blueberry anthocyanin extraction and purification method |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105510319A (en) * | 2015-12-07 | 2016-04-20 | 湖南科技大学 | Method for quickly identifying color of seed coats of rape seeds in early development period |
| CN106749151A (en) * | 2016-11-28 | 2017-05-31 | 沈阳农业大学 | A kind of rapid extracting method of blueberry pomace anthocyanidin |
| CN106749151B (en) * | 2016-11-28 | 2020-02-07 | 沈阳农业大学 | Method for rapidly extracting blueberry pomace anthocyanin |
| CN106699719A (en) * | 2016-11-29 | 2017-05-24 | 蓬莱海洋（山东）股份有限公司 | Method for efficiently extracting procyanidine ingredient from young apple fruits removed by thinning |
| CN106699719B (en) * | 2016-11-29 | 2019-04-09 | 蓬莱海洋（山东）股份有限公司 | A kind of apple removes young fruit procyanidine component highly effective extraction method |
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