CN102344470A - Method for extracting wild persimmon leaf tannin by ultrasonic wave auxiliary semi-bionic method - Google Patents
Method for extracting wild persimmon leaf tannin by ultrasonic wave auxiliary semi-bionic method Download PDFInfo
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Abstract
The invention relates to a method for extracting tannin from wild persimmon leaves by an ultrasonic wave auxiliary semi-bionic method, which is mainly characterized in that the de-enzymed wild persimmon leaves are dried and smashed, ethanol and water as used as extraction solvents, and backflow extraction is carried out 3 times at the temperature of 50-60 DEG C by simulating the pH value of a gastrointestinal tract; and in the process of each extraction, ultrasonic waves are adopted for auxiliary treatment; and extracts in three times are combined and centrifuged, and then the obtained extract is concentrated and dried in a vacuum mode at the temperature of 50-60 DEG C to obtain the wild persimmon leaf tannin. In the semi-bionic extraction method of the invention, ultrasonic wave auxiliary extraction is adopted to make wild persimmon leaf cell tissues be wall-broken or deformed so as to increase the tannin yield, shorten the extraction time and keep the activity of the tannin.
Description
Technical field
The present invention relates to a kind of method of extracting wild persimmon leaf tannin, particularly a kind of method of extracting wild persimmon leaf tannin with auxiliary half biomimetic method of ultrasonic wave.
Background technology
Tannin (Tannins); Also claim Weibull; Tannic acid; Be a kind of water-soluble, material that is present in the plant with polyphenol hydroxyl structure; Have effects such as the anti-oxidant of stronger biological activity, excellence and removing free radical, ability and metal ion-chelant, and can react with protein, alkaloid, polysaccharide etc.; Thereby show very strong resistance of oxidation, excellent antibiotic, antiviral activity and suppress carcinogens and tumor promotion.Demonstrate unique advantages at aspects such as preventing and treat free-radical induced disease, the peroxidation that suppresses lipid and other materials, AIDS resisting, be widely used in medicine, food, leather and printing and dyeing industry.
At present, the extraction of vegatable tannin mainly come from seeds such as black wattle plant skin, pine tree (referring to Northeast Forestry University's journal, 2009,37 (8): 67-70.) [U1].Wild persimmon tree distributes extensively in the area, the western Hunan in Hunan Province, all contain in wild persimmon Chinese olive and the leaf abundant persimmon tannin (referring to Journal of Ethno pharmacology, 2007,109:177-206.).Existing existing research about utilizing wild persimmon Chinese olive to extract tannin does not also appear in the newspapers but auxiliary half biomimetic method of relevant ultrasonic wave extracts wild persimmon leaf tannin.
Development along with fine chemistry industry; The ultrasonic-assisted extraction technology is being brought into play its special advantages in many aspects; Simultaneously; Semi-bionic extraction (SBE) then is a kind of novel plant extract technology that designs for bio-pharmaceuticals; The biological activity that adopts the guiding separation method under active the guidance and guarantee extract is (referring to Strait Pharmaceutical Journal; 2010,22 (4): 12-14.).
In order to make full use of wild persimmon resource, the present invention is a raw material with wild persimmon leaf, adopts semi-bionic extraction (SBE), and utilization ultrasonic wave ancillary technique is to obtain the high reactivity persimmon tannin.
China is the country of origin and the main product state of persimmon, is distributed in 23 provinces, cities and autonomous regions.At present, the extraction of persimmon tannin mainly is really to be raw material with blue or green persimmon.Below be really to be a series of progress situations of research object with persimmon both at home and abroad in recent years:
Domestic present Research has:
Gu Haifeng etc. have inquired into the extraction process of total phenol and condensed tannin in the flesh of persimmon; Adopt the Vanillin and the salt acid system of Folin-Denis method, improvement to carry out total phenol and condensed tannin Determination on content respectively; The anhydrous methanol of having confirmed to contain 1% hydrochloric acid is that the optimum extraction solvent is (referring to food science and technology; 2007,2:133-136; Transactions of the Chinese Society of Agricultural Engineering, 2007,23 (5): 241-245.); People such as Yang Wenkun discover persimmon tannin purifying process and antiallergic activity thereof: when the concentration of persimmon tannin reaches 5mg/mL; Its inhibiting rate to Unidasa can reach 95% (referring to the biological journal of mountain farming; 2010,29 (1): 48-51.); Chen Meihong etc. remove relatively studying of free radical effect to persimmon different sites tanning extract; Find persimmon skin, persimmon meat, the calyx and receptacle of a persimmon and the peaceful extract of persimmon vouching to hexichol for bitter taste free acyl radical (DPPH), hydroxyl radical free radical (OH), 2; 2'-joins the removing effect of nitrogen-two-(3-ethyl benzo thiazole phenanthroline-6-sulfonic acid) free radical (ABTS+) obviously (referring to food science and technology; 2009,34 (8): 120-123.); People such as Zhang Baoshan with sweet persimmon be controlled observation the OX-heart persimmon growth, take away the puckery taste and return puckery in the metamorphosis of tannin cell; Indicate puckery persimmon tannin cell vegetative period volume constantly increase, quantity is on the increase, astringent taste is strengthened (referring to Shaanxi Normal University's journal (natural science edition); 2008,36 (1): 99-105.); Yang Yongs etc. have done than have more comprehensively described to the form of persimmon fruit tannin cell on the basis of summing up forefathers' achievement, summed up 6 kinds of persimmons fruit tannin cells shape (referring to Xibei Univ. of Agricultural & Forest Science & Technology's journal, 2003,31 (6): 93-99; The northwest Botany Gazette, 2007,27 (8): 1524-1530.).
In addition; Zhang Jishu etc. take away the puckery taste to the whole bag of tricks, and tannin and persimmon Study on Hardness show in the persimmon of back, in the persimmon variation of solubility tannin content and persimmon changes in hardness similar (referring to the northwest Botany Gazette, the dry fruit progress; 2001,23 (2): 283-287.); People such as Zhang Haisheng research dried persimmon processing return puckery mechanism the time think under the effect of tannin in the persimmon at ethanol dehydrogenase; Converted products can not return puckery; But in the product after pyroprocessing; It is that reversible is (referring to foodstuffs industry science and technology that persimmon tannin forms compound state; 2003,24 (12): 39-40.).Scholars such as Dong Xinhai also form persimmon fruit tannin and biological characteristics is summarized and sum up.
The foreign study present situation has
Nineteen twenty-three Japan scholar Komatsu and Matsunami have just carried out preliminary study to persimmon fruit tannin; From immature puckery persimmon juice, extract a kind of aldehydes matter; And infer possibly contain in the tannin phloroglucin and gallic acid monomer (referring to NATURE, 1958,11:1371.).
Ito and Oshima find when research persimmon fruit tannin molecular formula in 1962; Its main component is named as persimmon fruit plain (diospyrin); Have a pyrocatechol and a pyrogallol; And the ortho position phenolic hydroxyl group of pyrocatechol and pyrogallol is easy to oxidized; Thereby can consume the oxygen in the environment; And free radicals such as ability assimilating activity oxygen (referring to garden examination newspaper, 1962, B (1): 1-16.).Simultaneously, the 70% ethanolic soln extract of discovering puckery persimmon of Katsube T. etc. have very strong anti-LDL oxidation capacity (referring to Agric Food Chem, 2004,52 (8): 2391-2396.).
People such as Satoshi Taira are in the period of 1997 to 1998; Studied the influence of fruit soluble pectin of persimmon in the deastringency of persimmon process and freezing rate to deastringency of persimmon; Discovery is in mixing solutions; In the persimmon fruit soluble pectin matter than white protein more easily with persimmon fruit tannin formation stable compound; Be beneficial to taking away the puckery taste of persimmon fruit (referring to Postharvest Biology and Technology; 1998,14 (3): 317-324; Postharvest Biology and Technology, 1997,12 (3): 265-271.).
Salvador research persimmon fruit finds that the cell that do not take away the puckery taste is irregular shape during cellularstructure, combines closely, and cytolemma and the wall that takes away the puckery taste degrade gradually, loose.The shape that shows tannin content and cell has certain relation (referring to Postharvest Biology and Technology, 2007, (12): 12-16.).
In addition, except the above-mentioned relevant research of persimmon tannin, many scholars also study other vegatable tannin, as scholars such as Li Min to different varieties tealeaves tannin carried out studying (referring to Food science, 2009,30 (2): 149-153.); Profound Nian Yong etc. to maletto tannin carried out analyzing (referring to the biomass chemical engineering, 2009,43 (5): 39-41.); Scholars such as Zhang Jian are with ultrasonic applications tannin in extracting stem of Fructus Vitis viniferae; Chen Xiaojuan and Li Li etc. carried out the research that utilizes half biomimetic method to extract chlorogenic acid and baicalin in the Folium Eucommiae (referring to fine chemistry industry, 2006,23 (3): 35-38; Chinese patent medicine, 2007,29 (10): 1527-1529.).
In sum; Chinese scholars and R&D institution have had realized that the significance of from biological material, extracting the vegatable tannin of high biological activity; And this has been launched some researchs; But the research report that from plant leaf, extracts tannin is also few; And relevant " research that auxiliary half biomimetic method of ultrasonic wave extracts wild persimmon leaf tannin technology " does not also have relevant report; This research will be carried out the discussion of this respect as point of penetration, for from leaf, extracting effective ingredients in plant a research direction will be provided.
The main method of preparation persimmon tannin is the water-bath circumfluence method at present, promptly takes different organic solvents that the tannin in the raw material is dissolved, and again lysate is concentrated and processes.
Mainly there is following deficiency in the water-bath circumfluence method when extracting persimmon tannin: complex process, and the tannin yield is low; Extraction time is long, needs through multiple extraction, and extraction cost is high; By product is many in the product, separate complex; The activity of tannin is lower, should not process biotechnological formulation, has influenced the purposes aspect bio-pharmaceuticals; Owing to used a large amount of organic solvents, caused the pollution of VOC easily.
Summary of the invention
The object of the present invention is to provide a kind of method of from wild persimmon leaf, extracting tannin with auxiliary half biomimetic method of ultrasonic wave.
Technical scheme of the present invention is: a kind ofly extract the method for wild persimmon leaf tannin with auxiliary half biomimetic method of ultrasonic wave, it is characterized in that: may further comprise the steps:
1. the wild persimmon leaf after will completing is dry, be crushed to 30 orders; Press with ethanol and water after the mixed of 1:1.5-1:3 as extracting solvent; The pH value of Gl tract; Regulate the pH value respectively and be 2.0-2.2,7.0-7.5,8.0-8.3; Extract 3 times 50-60 ° of C temperature refluxed, extracted 30-40 minute at every turn;
2. under step extraction conditions 1., in the process of extracting, adopt ultrasonic wave to carry out auxiliary treatment, the treatment time is 15-20 minute;
3. extract the gained extracting solution with 3 times and merge,, remove out residue,, promptly obtain wild persimmon leaf tannin dry under 50-60 ° of C temperature behind the parting liquid process vacuum concentration that obtains with the centrifugal 30min of 4500r/min.
The pH value that refluxing extraction of the present invention is 3 times is adjusted to 2.0,7.2,8.0 respectively.
The pH value that refluxing extraction of the present invention is 3 times is adjusted to 2.1,7.4,8.1 respectively.
The pH value that refluxing extraction of the present invention is 3 times is adjusted to 2.2,7.5,8.2 respectively.
Hyperacoustic power of the present invention is that 200-300 W, frequency are 26-32 kHz.
The usage quantity of ethanol of the present invention and water extraction solvent and the ratio control of whole consumption are at 30%-40%.
The distinguishing feature of technical scheme of the present invention is in half biomimetic method leaching process, to adopt ultrasonic-assisted extraction; Utilize multistage physics usefulness such as hyperacoustic cavitation, mechanical vibration, microjet and little acoustic streaming; Impel wild persimmon tree leaf cell organize broken wall or distortion improve tannin yield, reduce extraction time, and keep the activity of tannin.
Preferred extraction solvent---the ratio of ethanol and water: 1:1.5-1:3 is disclosed among the present invention, and the pH value of extracting solution.
The invention also discloses hyperacoustic power, frequency and action time in the half biomimetic method leaching process.Auxiliary half biomimetic method of ultrasonic wave extracts wild persimmon leaf tannin, and the second alcohol and water that adopts 1:1.5-1:3 extracts 3 times for extracting solvent, and 3 times pH value is respectively 2.0,7.2,8.0 or 2.1,7.4,8.1 or 2.2,7.5,8.2; Simultaneously; Hyperacoustic power is that 200-300 W, frequency are that 26-32 kHz, treatment time are 15-20 min; Pass through vacuum concentration again after centrifugal 30 min processing separates residue through 4500 r/min with resulting extracting solution; And dry under the low temperature of 50-60 ° of C, promptly obtain wild persimmon leaf tannin.
The basic parameters such as content of wild persimmon leaf tannin detect according to People's Republic of China's forestry industry standard " LY/T1642-2005 " and " LY/T1300-2005 ".
The present invention is controlled at single extraction time 30-40 minute; The usage quantity of ethanol and water extraction solvent and the ratio control of whole consumption have shortened extraction time, have reduced the use of extracting solvent, the activity that has kept tannin in 25%-40% in leaching process.
Because the composition more complicated of persimmon tannin, in leaching process, receive the influence of external conditionss such as heating, illumination easily and variation such as oxidation, isomery takes place and influence quality product.Adopt half biomimetic method technology can reduce the extraction temperature, prevent the oxidation of tannin under the high temperature, if but the time of extract at low temperature oversizely equally also can cause oxidation because of long-time the contact with air.Use ultrasonic-assisted extraction, then can shorten extraction time, can not produce excessive influence, can better keep Physiological activities of tannin, have application promise in clinical practice its molecular structure and physico-chemical property.Simultaneously, hyperacoustic use can also improve the yield of tannin, so can reduce cost.
In addition, extract suitable use ethanol in the solvent, can improve the solubleness of tannin and help extracting, and ethanol the later stage tannin concentrate with drying process in can reclaim through distillation, so do not have environmental pollution in the whole leaching process, and simple to operate.
Use the resulting tannin of method provided by the invention to detect with ultraviolet spectrophotometer; Wherein NVP-XAA 723 (EGCG), L-Epicatechin gallate (ECG), epigallocatechin are formed; Total content reaches 95.6%, can directly be used for food, medicine and other fields use.
Embodiment
Through specific embodiment the present invention is described further below.
Used major equipment of the embodiment of the invention and instrument have:
JY92-II D type ultrasonic cell disruptor (NingBo XinZhi Biology Science Co., Ltd); 722 type ultraviolet spectrophotometers (Shanghai Precision Scientific Apparatus Co., Ltd); SHZ-82 digital display constant temperature water bath vibrator (Jiangsu section is emerging); The FW117 herbal medicine is pulverized machine (Tianjin Tai Site Instr Ltd.); TDL-40B whizzer (U.S. Anke company).
Embodiment 1
This instance adopts and extracts for 3 times, then extracting solution is mixed and concentrate drying.
Extract for the first time: accurately take by weighing and complete after drying and be ground into the wild persimmon leaf of 30 purposes powder 100g; Add 300g and extract solvent (mass ratio of ethanol and water is 1:1.5); Using the second acid for adjusting pH value is 2.0; Heating in water bath to 50 ° C; Insulation was extracted 40 minutes; It is that the ultrasonic wave of 26 kHz assists half biomimetic method to extract as 200W, frequency that power was used at every during this time interval in 5 minutes, and be 5 minutes each duration of service, amounts to 20 minutes.Obtain containing the extracting solution of tannin after the filtration.
Extract for the second time: add 200g in the wild persimmon leaf residue after above-mentioned filtration and extract solvent (mass ratio of ethanol and water is 1:2); Using yellow soda ash to regulate the pH value is 7.2; Heating in water bath to 50 a ° C begins insulation and extracted 40 minutes; It is that the ultrasonic wave of 26 kHz assists half biomimetic method to extract as 200W, frequency that power was used at every during this time interval in 5 minutes; Time is 5 minutes, amounts to 20 minutes.Obtain containing the extracting solution of tannin after the filtration.
Extract for the third time: in the wild persimmon leaf residue that extracts for the second time after filtering, add 200g and extract solvent (mass ratio of ethanol and water is 1:3); Using yellow soda ash to regulate the pH value is 8.0; Heating in water bath to 50 a ° C begins insulation and extracted 30 minutes; It is that the ultrasonic wave of 26 kHz assists half biomimetic method to extract as 200W, frequency that power was used at every during this time interval in 5 minutes; Time is 5 minutes, amounts to 15 minutes.Obtain containing the filtration extracting solution of tannin after the filtration.
The extracting solution that obtains for above-mentioned 3 times is mixed, after centrifugal 30 min handle through 4500 r/min, the parting liquid that obtains is promptly obtained wild persimmon leaf tannin after vacuum concentration, drying under 50 ° of C temperature.
Embodiment 2
This instance adopts and extracts for 3 times, then extracting solution is mixed and concentrate drying.
Extract for the first time: accurately take by weighing and complete after drying and be ground into the wild persimmon leaf of 30 purposes powder 100g; Add 300g and extract solvent (mass ratio of ethanol and water is 1:1.5); Using the second acid for adjusting pH value is 2.1; Heating in water bath to 55 ° C; Insulation was extracted 40 minutes; It is that the ultrasonic wave of 29 kHz assists half biomimetic method to extract as 250W, frequency that power was used at every during this time interval in 10 minutes, and be 10 minutes each duration of service, amounts to 20 minutes.Obtain containing the extracting solution of tannin after the filtration.
Extract for the second time: add 200g in the wild persimmon leaf residue after above-mentioned filtration and extract solvent (mass ratio of ethanol and water is 1:2); Using yellow soda ash to regulate the pH value is 7.4; Heating in water bath to 55 a ° C begins insulation and extracted 40 minutes; It is that the ultrasonic wave of 29 kHz assists half biomimetic method to extract as 250W, frequency that power was used at every during this time interval in 10 minutes; Time is 5 minutes, amounts to 15 minutes.Obtain containing the extracting solution of tannin after the filtration.
Extract for the third time: in the wild persimmon leaf residue that extracts for the second time after filtering, add 200g and extract solvent (mass ratio of ethanol and water is 1:3); Using yellow soda ash to regulate the pH value is 8.1; Heating in water bath to 55 a ° C begins insulation and extracted 30 minutes; It is that the ultrasonic wave of 29 kHz assists half biomimetic method to extract as 250W, frequency that power was used at every during this time interval in 5 minutes; Time is 5 minutes, amounts to 15 minutes.Obtain containing the extracting solution of tannin after the filtration.
The extracting solution that obtains for above-mentioned 3 times is mixed, after centrifugal 30 min handle through 4500 r/min, the parting liquid that obtains is promptly obtained wild persimmon leaf tannin after vacuum concentration, drying under 60 ° of C temperature.
Embodiment 3
This instance adopts and extracts for 3 times, then extracting solution is mixed and concentrate drying.
Extract for the first time: accurately take by weighing and complete after drying and be ground into the wild persimmon leaf of 30 purposes powder 100g; Add 300g and extract solvent (mass ratio of ethanol and water is 1:3); Using the second acid for adjusting pH value is 2.2; Heating in water bath to 60 ° C; Insulation was extracted 40 minutes; It is that the ultrasonic wave of 32 kHz assists half biomimetic method to extract as 300W, frequency that power was used at every during this time interval in 10 minutes, and be 10 minutes each duration of service, amounts to 20 minutes.Obtain containing the extracting solution of tannin after the filtration.
Extract for the second time: add 200g in the wild persimmon leaf residue after above-mentioned filtration and extract solvent (mass ratio of ethanol and water is 1:2); Using yellow soda ash to regulate the pH value is 7.5; Heating in water bath to 60 a ° C begins insulation and extracted 40 minutes; It is that 300 W, frequency are that the ultrasonic wave of 32 kHz assist the extraction of half biomimetic method that power is used at every during this time interval 10 minutes; Time is 10 minutes, amounts to 20 minutes.Obtain containing the extracting solution of tannin after the filtration.
Extract for the third time: in the wild persimmon leaf residue that extracts for the second time after filtering, add 200g and extract solvent (mass ratio of ethanol and water is 1:3); Using yellow soda ash to regulate the pH value is 8.2; Heating in water bath to 60 a ° C begins insulation and extracted 30 minutes; It is that the ultrasonic wave of 32 kHz assists half biomimetic method to extract as 300W, frequency that power was used at every during this time interval in 5 minutes; Time is 5 minutes, amounts to 15 minutes.Obtain containing the extracting solution of tannin after the filtration.
The extracting solution that obtains for above-mentioned 3 times is mixed, after centrifugal 30 min handle through 4500 r/min, the parting liquid that obtains is promptly obtained wild persimmon leaf tannin after vacuum concentration, drying under 60 ° of C temperature.
Claims (6)
1. one kind is extracted the method for wild persimmon leaf tannin with auxiliary half biomimetic method of ultrasonic wave, it is characterized in that: may further comprise the steps:
1. the wild persimmon leaf after will completing is dry, be crushed to 30 orders; With ethanol and water in the ratio of 1:1.5-1:3 as extracting solvent; The pH value of Gl tract; Regulate the pH value respectively and be 2.0-2.2,7.0-7.5,8.0-8.3; Extract 3 times 50-60 ° of C temperature refluxed, extracted 30-40 minute at every turn;
2. under step extraction conditions 1., in the process of extracting, adopt ultrasonic wave to carry out auxiliary treatment, the treatment time is 15-20 minute;
3. extract the gained extracting solution with 3 times and merge,, remove out residue,, promptly obtain wild persimmon leaf tannin dry under 50-60 ° of C temperature behind the parting liquid process vacuum concentration that obtains with the centrifugal 30min of 4500r/min.
2. method according to claim 1 is characterized in that: the pH value that refluxing extraction is 3 times is adjusted to 2.0,7.2,8.0 respectively.
3. method according to claim 1 is characterized in that: the pH value that refluxing extraction is 3 times is adjusted to 2.1,7.4,8.1 respectively.
4. method according to claim 1 is characterized in that: the pH value that refluxing extraction is 3 times is adjusted to 2.2,7.5,8.2 respectively.
5. method according to claim 1 is characterized in that: hyperacoustic power is that 200-300 W, frequency are 26-32 kHz.
6. method according to claim 1 is characterized in that: the usage quantity of ethanol and water extraction solvent and the ratio control of whole consumption are at 30%-40%.
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| CN103245551A (en) * | 2013-05-24 | 2013-08-14 | 西南大学 | Method for quickly determining tannin content in mulberry leaves |
| CN107912774A (en) * | 2017-12-11 | 2018-04-17 | 深圳市葵瑞生命科技发展有限公司 | A kind of tender okra pod extract preparation process |
| CN109568993A (en) * | 2018-11-05 | 2019-04-05 | 广州柿宝生物科技有限公司 | A kind of dehydration and drying technique of persimmon tannin concentrate |
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| CN103245551A (en) * | 2013-05-24 | 2013-08-14 | 西南大学 | Method for quickly determining tannin content in mulberry leaves |
| CN107912774A (en) * | 2017-12-11 | 2018-04-17 | 深圳市葵瑞生命科技发展有限公司 | A kind of tender okra pod extract preparation process |
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Application publication date: 20120208 |