CN102382020A - Method for simultaneously extracting alliin and garlic enzyme from garlic - Google Patents
Method for simultaneously extracting alliin and garlic enzyme from garlic Download PDFInfo
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- CN102382020A CN102382020A CN2010102677579A CN201010267757A CN102382020A CN 102382020 A CN102382020 A CN 102382020A CN 2010102677579 A CN2010102677579 A CN 2010102677579A CN 201010267757 A CN201010267757 A CN 201010267757A CN 102382020 A CN102382020 A CN 102382020A
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Abstract
The invention relates to a method for simultaneously extracting alliin and garlic enzyme from garlic, which comprises: full ultrasonic cell wall breakage of mixture of peeled garlic and a polar organic solvent with the mass being at least four times of the mass of the garlic (pharmaceutically acceptable) and the volumetric water content less than or equal to 10 percent under the conditions with the pH being 5.5-6.8 and the temperature being less than or equal to 35 DEG C, solid-liquid separation, filtration and separation of supernatant by molecule ultrafiltration membranes with the molecular weight reducing gradually, wherein the molecular weight of the molecule ultrafiltration membrane at the last stage is 10,000 to 3,000, concentration of the filtrate by a nano filtration membrane, crystallization of the concentrated solution, separation to obtain crystals, and drying under conditions with the temperature less than or equal to 35 DEG C. The problem of reaction of the alliin and the garlic enzyme in cell wall breakage of the garlic can be solved effectively, and garlic extracts with the alliin content being 2.2-98 percent and the activity ratio of the garlic enzyme being 80-900 can be extracted and enriched, the losses of effective ingredients can be reduced, the yield and purity can be improved, the cost is low, the production requirements of cleanness and low energy consumption can be satisfied, and the method has a practical value in industrial applications.
Description
Technical field
The present invention relates to a kind of preparation method of Bulbus Allii extract, specifically is that a kind of can the extraction simultaneously obtains alliin and two kinds of extraction of effective components of allinase.
Background technology
Garlic (Aliium Sativm) is the bulb of the living garlic of Liliaceae allium, is a kind of plant with medicine-food two-purpose function, and the good reputation of " natural medicinal plant gold " is arranged.Research shows, garlic have sterilization, antiviral, reducing blood-fat, antithrombotic, antitumor, regulate multiple efficacies such as human immunity.Therefore the research to the activeconstituents of garlic becomes both at home and abroad the focus of research for many years.Alliin is one of main biologically active substance in the garlic, is non-protein sulfur-containing amino acid unique in the garlic, content 0.6%-2% in the garlic dry-matter.Allinase is a kind of endogenous enzyme that is present in the garlic, is a kind of dimer gp of homogeneous, accounts for the 10-20% of soluble proteins in the garlic.
In garlic, alliin and allinase are in the different sites of garlic cell respectively.Alliin is present in the tenuigenin with stable form, and allinase then is present in the cell vacuole.After garlic received the external force effect that its cell is destroyed, enzyme digestion reaction promptly can take place with after alliin contacts in allinase, generated allicin.Allicin can generate Diallyl Thiosulfonate (being garlicin) under the further effect of enzyme.Diallyl Thiosulfonate is only the final effective constituent product that produces above-mentioned beneficial functional.And allicin, Diallyl Thiosulfonate all are the moment products, can't exist under normal conditions.Alliin and allinase can be stablized each other and not react under the state of moisture＜8%.And enzymolysis under the effect of the back water in the human body stomach that is eaten, the Diallyl Thiosulfonate that moment produces only just maybe the strong effect of tool under the rapid absorbing state by stomach.This also is that Oleum Bulbus Allii and the long-term edible garlic of people and being difficult to obtains and the major reason of effect.
Because when the garlic composition is extracted; Alliin and allinase two compositions can react when the garlic cell wall breaking and become a pair of paradox; Though therefore from garlic, extract the at present existing a lot of bibliographical informations of the method for alliin, allinase respectively; All be to adopt chemical process, and mostly only limit to the laboratory, do not possess industrial application its single separation.And these two kinds of compositions quilts are exactly two common separate constituents after independent the separation, all can not possess the exclusive medical treatment and health care value of garlic.
Summary of the invention
To above-mentioned situation, the invention provides a kind of new preparation process that obtains Bulbus Allii extract, particularly can extract the process for extracting that obtains alliin and two kinds of effective constituents of allinase and it is reacted simultaneously.
The present invention extracts the method for alliin and allinase simultaneously from garlic, be with the peeling garlic be at least 4 times of its quality, be preferably 4～12 times; Be more preferably and accept in 5～9 times the pharmacy and the mixture of the polar organic solvent of volumetric(al) moisture content≤10%; Be adjusted under ℃ condition of pH5.5～6.8 and temperature≤35, mixing the making beating broken wall, after effective constituent fully being soaked dissolve with UW; Solid-liquid separation (for example can adopt centrifugal or sheet frame mode commonly used to filter); The clear liquid that obtains separates through the descending molecule ultrafiltration membrance filter of molecular weight successively, and the molecular weight of final stage molecule ultra-filtration membrane is 10000～3000, and filtrating is again after nanofiltration membrane concentrates; The liquid concentrator convection drying obtains crystal or is gone ahead of the rest to separate behind the crystallization by liquid concentrator again obtaining crystal, and crystal is dry under≤35 ℃ of conditions.
Research shows, carries out broken wall with ultrasonic power, can improve the extraction leaching yield (>=50%) of garlic composition greatly.Be present in effective constituent alliin and allinase in garlic tenuigenin and the cell vacuole respectively, after the garlic cell was destroyed, the two was in contact with one another and can reacts rapidly, produced allicin, and further reaction is Diallyl Thiosulfonate.The polar organic solvent that uses anhydrous or water cut≤10% is as extracting solvent, and controls the pH value scope of material system simultaneously, the effective activity of passivation allinase, thus can effectively control the reaction of alliin and allinase behind the garlic cell wall breaking.
Can accept in the above-mentioned said pharmacy and the polar organic solvent of volumetric(al) moisture content≤10%, serve as preferred with solvent the most commonly used such as methyl alcohol, ethanol, acetone generally, and wherein best is the employing absolute ethyl alcohol.
Above-mentioned pH value with the supersonic wave wall breaking material handling is regulated, and generally can select acetic acid commonly used in the medicine, Hydrocerol A etc.Wherein, more preferred pH value scope is 6.0～6.5.
Test shows, the garlic material carried out supersonic wave wall breaking when handling, and generally adopting frequency is that the supersonic wave wall breaking of 20～80KHz was handled 10～45 minutes, promptly can obtain satisfied shell-broken effect, the cell wall breaking rate can >=95%.When carrying out supersonic wave wall breaking, can carry out the mode of tracing observation, the broken wall process is controlled through pair cell broken wall situation under the powerful microscope.Wherein, the preferred temperature condition of broken wall treatment is 20 ℃～35 ℃, helps avoiding the decomposition of alliin and allinase as far as possible.
Garlic liquid material behind the broken wall is after solid-liquid separation, and filtrating is used the descending molecule ultra-filtration membrane of molecular weight filtering separation successively again, can make the alliin that is dissolved in behind the broken wall in the solvent and allinase and other unwanted component separating and obtains purifying.
According to demands of different to active constituent content in the products therefrom; When carrying out separation and purification with the molecule ultra-filtration membrane; The molecular weight ranges of used molecule ultra-filtration membrane and/or filtering progression (number of times); Can adopt different flexible selections such as one or more levels different molecular weight molecule ultra-filtration membrane, test shows that the filtrating after the solid-liquid separation is generally carried out 2～3 grades of filtrations successively with the molecule ultra-filtration membrane and got final product; The maximum molecular weight that wherein filters the molecule ultra-filtration membrane that uses first can be 150000, and the molecule ultra-filtration membrane molecular weight that last uses should be 10000～3000.With three-stage filtration separation and purification mode is example, and the molecular weight of the molecule ultrafiltration ripple of the general first step can be 150,000～20,000, and the film of 20,000～10,000 molecular weight can be adopted in the second stage, and the third stage then can be adopted the film of 10,000～3,000 molecular weight).Said molecule ultrafiltration waveshape can adopt industry to go up rolled film commonly used or tubular membrane etc., and concrete material can be tubular fibre, engineering plastics or pottery etc.In order to prevent the intensification in ultra-filtration process, the heat exchanger that can be used appropriate form is lowered the temperature.
Filtrating after the separation and purification of molecule ultra-filtration membrane suitably concentrates through nanometer film again, can help next step crystallization and raising crystalline yield.Generally can select to adopt molecular weight commonly used is that 100～200 tubular type or rolling nanometer film concentrate, and temperature should not surpass 60 ℃ equally, is preferably carrying out below 30 ℃.Improve spissated ratio, the crystallization that can help next step reaches resultant crystalline dry.Test shows, preferred concentrating degree is that to make the proportion of liquid concentrator be 1.05～1.15.
Filtrating after nanofiltration membrane concentrates through cooling, can have the said crystal that contains alliin and allinase two compositions simultaneously to separate out.Abundant for helping crystallization, improve yield.For example; For preparing 50 (w) % above high purity alliin and the allinase of specific activity more than 500; Crystallization should carry out under-3 ℃～0 ℃ freezing conditions; And can carry out recrystallization again with same solvent and condition to the gained crystallisate, can get content and be 50～99% the alliin and the allinase crystal of specific activity 500～900.
As above-mentioned, temperature raises and can quicken the decomposition of alliin and allinase, therefore should not be above 60 ℃ to the crystalline drying temperature that obtains behind the crystallization.Test shows, the optimal way of a kind of thorough drying that can be for reference is, being 20 ℃～35 ℃ in temperature is to carry out microwave vacuum drying under the condition of 0.08～0.09MPa with vacuum tightness, and the microwave frequency that can select is 2200～3200MHz.
Said extracted method of the present invention; Effectively and solved problem and the contradiction that two kinds of effective constituent alliins and allinase in the garlic contact and can react satisfactorily when cell wall breaking; Can extract and obtain being enriched with alliin content simultaneously and be 2.2～98% with the Bulbus Allii extract of allinase specific activity 80～900; Loss of active ingredients is few, and yield and purity are high.The whole process of this method remains on carries out under the lower temperature and accomplishes; Both avoided alliin and allinase >=60 ℃ of inducible decomposition, and realized the production requirement of less energy-consumption, it is low to have reduced cost; Realization cleans production, has practical application in industry and is worth.
Below again foregoing of the present invention is done further to specify through the embodiment of embodiment.But should this be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following instance.Do not breaking away under the above-mentioned technological thought situation of the present invention, various replacements or change according to ordinary skill knowledge and customary means are made all should comprise within the scope of the invention.
Embodiment
Embodiment 1
Get 100 kilograms of fresh garlics, distinguish, decortication are with 500 kilograms of 99% ethanol; Add 0.5kg lemon acid for adjusting pH 6.0～6.5, the UW with frequency 26～30KHz mixed the making beating broken wall treatment 45 minutes, centrifuging at normal temperatures; The tubular ultra-filtration membrane of filtrating through 20,000 molecular weight separates, and parting liquid concentrates with the nf membrane of molecular weight 100, to liquid concentrator proportion be 1.08～1.1; Directly under frequency 2300MHz and vacuum tightness 0.08MPa condition, carry out microwave vacuum drying in 28 ℃.11.7 kilograms of Bulbus Allii extracts that contained alliin content 10.2% and allinase specific activity 200～300 simultaneously.
Embodiment 2
Take from 100 kilograms in right exsiccant garlic, distinguish, decortication add 1.5 kilograms of alimentary acetic acids in 900 kilograms of 90% ethanol; Regulate pH to 6.0～6.5; With the UW of frequency 26～30KHz, under 30 ℃ of conditions, mix the making beating broken wall treatment after 20 minutes, solid-liquid separation; Filtrating is with tubular ultra-filtration membrane classified filtering separation and purification successively, and the molecular weight of filter membrane is respectively 50000,10000,5000.Parting liquid is concentrated into about 50 liters with the nf membrane of molecular weight 100～200 ,-3 ℃ of freezing crystallizatioies.After the solid-liquid separation, concentrate-3 ℃ of frozen recrystallizations with 5-9 times of absolute ethyl alcohol normal-temperature dissolution and same nf membrane again.After the crystal separation, microwave vacuum drying (2600MHz, 0.09MPa), 0.75 kilogram of alliin, allinase specific activity 500～600 Bulbus Allii extract of purity 91.5%.Whole process is produced each process section controlled temperature below 30 ℃.
Embodiment 3
Get 100 kilograms in the garlic of refrigeration air adjusting storeroom, distinguish, decortication, 95% methyl alcohol adds 1 kilogram of Hydrocerol A for 600 kilograms.Regulate pH to 6.0～6.5, UW (frequency 50KHz) mixes making beating, and broken wall extracted 30 minutes; After the Plate Filtration solid-liquid separation; Filtrating is separated through the ultra-filtration membrane grading purification of 20000,5000 molecular weight successively, and parting liquid is concentrated into proportion about 1.1 through the nf membrane of molecular weight 100～200 again, in 30 ℃ of direct microwave vacuum drying (3000KHz; 0.08MPa), obtain containing alliin content 20.6% and 5.8 kilograms of allinase specific activity 600～700 Bulbus Allii extracts.
Claims (10)
1. from garlic, extract the method for alliin and allinase simultaneously; It is characterized in that the garlic of peeling can be accepted in the pharmacy of 4 times of its quality and the polar organic solvent of volumetric(al) moisture content≤10% with being at least; Be adjusted under ℃ condition of pH value 5.5～6.8 and temperature≤35 behind the abundant broken wall of UW, solid-liquid separation, clear liquid separates through the descending molecule ultrafiltration membrance filter of molecular weight successively; The molecular weight of final stage molecule ultra-filtration membrane is 10000～3000; After nanofiltration membrane concentrated, the liquid concentrator crystallization separated obtaining crystal and drying under≤35 ℃ of conditions to filtrating again.
2. process for extracting as claimed in claim 1, the consumption that it is characterized in that said polar organic solvent are 4～12 times of garlic quality.
3. process for extracting as claimed in claim 2, the consumption that it is characterized in that said polar organic solvent are 5～9 times of garlic quality.
4. process for extracting as claimed in claim 1 is characterized in that said polar organic solvent is ethanol or methyl alcohol.
5. process for extracting as claimed in claim 1 is characterized in that the material pH value when said supersonic wave wall breaking is handled is 6.0～6.5.
6. process for extracting as claimed in claim 1 is characterized in that the ultrasonic frequency that said supersonic wave wall breaking is handled is 20～80KHz, and the broken wall treatment time is 10～45 minutes.
7. process for extracting as claimed in claim 1, it is characterized in that said supersonic wave wall breaking is handled carries out under 20 ℃～35 ℃ temperature condition.
8. process for extracting as claimed in claim 1; It is characterized in that the filtrating after the said solid-liquid separation carries out 2～3 grades of filtrations successively with the molecule ultra-filtration membrane; Filter the molecule ultra-filtration membrane that uses first and be molecular weight 1 50000, the molecular weight of the molecule ultra-filtration membrane that last uses is 10000～3000.
9. process for extracting as claimed in claim 1 is characterized in that said liquid concentrator freezing crystallization under-3 ℃～0 ℃ condition.
10. like the described process for extracting of one of claim 1 to 9, it is characterized in that to the crystal that obtains behind the crystallization using frequency is the microwave vacuum drying of 2200～3200MHz under 20 ℃～35 ℃ of temperature and vacuum tightness 0.08～0.09MPa condition.
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| CN2010102677579A CN102382020A (en) | 2010-08-31 | 2010-08-31 | Method for simultaneously extracting alliin and garlic enzyme from garlic |
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| CN2010102677579A CN102382020A (en) | 2010-08-31 | 2010-08-31 | Method for simultaneously extracting alliin and garlic enzyme from garlic |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102697048A (en) * | 2012-06-11 | 2012-10-03 | 邯郸市春风科技发展有限公司 | Method for extracting natural active substance in garlic by using ultrasonic wave |
| CN102850245A (en) * | 2012-09-04 | 2013-01-02 | 江苏大学 | Method for ultrasonic assisted extraction of alliin in garlic |
| CN104059002A (en) * | 2014-06-27 | 2014-09-24 | 齐鲁工业大学 | Ultrasonic assisted aqueous two-phase extraction method for alliin |
| CN104087627A (en) * | 2014-07-29 | 2014-10-08 | 江南大学 | Method for preparing gram-level high-purity natural thiosulfinate |
| CN105646301A (en) * | 2016-02-02 | 2016-06-08 | 陈虎 | Method for extracting alliin and co-producing garlic polysaccharide and garlic powder from garlic |
| CN106819963A (en) * | 2016-12-08 | 2017-06-13 | 四川小叶本草生物科技有限公司 | One kind protection alliin, allinnase（Allinase）The method of active component |
| CN107177579A (en) * | 2017-05-17 | 2017-09-19 | 江南大学 | A kind of method that utilization garlic pieces processing waste water prepares allinnase, allicin and garlic polysaccharide |
| CN107653232A (en) * | 2017-11-14 | 2018-02-02 | 合肥创想能源环境科技有限公司 | A kind of technique of ultrafiltration purifying concentration garlic superoxide dismutase |
| CN108434307A (en) * | 2018-03-09 | 2018-08-24 | 新疆埃乐欣药业有限公司 | Antipruritic anti-acne garlic ointment of inhibiting bacteria and diminishing inflammation and its preparation method and application |
| CN108913678A (en) * | 2018-07-23 | 2018-11-30 | 江苏挺卫实业有限公司 | A kind of separating and extracting process of the garlic allinase for cereal beverage |
| CN112707852A (en) * | 2020-12-25 | 2021-04-27 | 青岛博恩高科生物技术有限公司 | Combined preparation method of garlic extract |
| CN114133346A (en) * | 2021-12-06 | 2022-03-04 | 无锡江大百泰科技有限公司 | Alliin and allicin combined extraction and preparation method |
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Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102697048A (en) * | 2012-06-11 | 2012-10-03 | 邯郸市春风科技发展有限公司 | Method for extracting natural active substance in garlic by using ultrasonic wave |
| CN102850245A (en) * | 2012-09-04 | 2013-01-02 | 江苏大学 | Method for ultrasonic assisted extraction of alliin in garlic |
| CN102850245B (en) * | 2012-09-04 | 2014-07-30 | 江苏大学 | Method for ultrasonic assisted extraction of alliin in garlic |
| CN104059002A (en) * | 2014-06-27 | 2014-09-24 | 齐鲁工业大学 | Ultrasonic assisted aqueous two-phase extraction method for alliin |
| CN104059002B (en) * | 2014-06-27 | 2016-08-17 | 齐鲁工业大学 | A kind of ultrasonic wave added aqueous two-phase extracts the method for alliin |
| CN104087627A (en) * | 2014-07-29 | 2014-10-08 | 江南大学 | Method for preparing gram-level high-purity natural thiosulfinate |
| CN105646301B (en) * | 2016-02-02 | 2018-05-01 | 陈虎 | From extracting alliin from garlic coproduction garlic polysaccharide and the method for garlic powder |
| CN105646301A (en) * | 2016-02-02 | 2016-06-08 | 陈虎 | Method for extracting alliin and co-producing garlic polysaccharide and garlic powder from garlic |
| CN106819963A (en) * | 2016-12-08 | 2017-06-13 | 四川小叶本草生物科技有限公司 | One kind protection alliin, allinnase（Allinase）The method of active component |
| CN107177579A (en) * | 2017-05-17 | 2017-09-19 | 江南大学 | A kind of method that utilization garlic pieces processing waste water prepares allinnase, allicin and garlic polysaccharide |
| CN107653232A (en) * | 2017-11-14 | 2018-02-02 | 合肥创想能源环境科技有限公司 | A kind of technique of ultrafiltration purifying concentration garlic superoxide dismutase |
| CN108434307A (en) * | 2018-03-09 | 2018-08-24 | 新疆埃乐欣药业有限公司 | Antipruritic anti-acne garlic ointment of inhibiting bacteria and diminishing inflammation and its preparation method and application |
| CN108913678A (en) * | 2018-07-23 | 2018-11-30 | 江苏挺卫实业有限公司 | A kind of separating and extracting process of the garlic allinase for cereal beverage |
| CN112707852A (en) * | 2020-12-25 | 2021-04-27 | 青岛博恩高科生物技术有限公司 | Combined preparation method of garlic extract |
| CN114133346A (en) * | 2021-12-06 | 2022-03-04 | 无锡江大百泰科技有限公司 | Alliin and allicin combined extraction and preparation method |
| CN114133346B (en) * | 2021-12-06 | 2022-07-29 | 无锡江大百泰科技有限公司 | Alliin and allicin combined extraction and preparation method |
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Application publication date: 20120321 |