CN104059002A - Ultrasonic assisted aqueous two-phase extraction method for alliin - Google Patents
Ultrasonic assisted aqueous two-phase extraction method for alliin Download PDFInfo
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- CN104059002A CN104059002A CN201410296457.1A CN201410296457A CN104059002A CN 104059002 A CN104059002 A CN 104059002A CN 201410296457 A CN201410296457 A CN 201410296457A CN 104059002 A CN104059002 A CN 104059002A
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Abstract
The invention belongs to the field of deep processing of garlic, and particularly discloses an ultrasonic assisted aqueous two-phase extraction method for alliin. The method comprises the steps of (1) adding inorganic salt into a Britton-Robinson buffer solution and dissolving, adding an organic solvent, uniformly mixing, standing and splitting the phases, so as to obtain an aqueous two-phase extractant; and (2) adjusting the pH of the aqueous two-phase extractant to be 2.0-4.0, adding garlic powder into the aqueous two-phase extractant, extracting assisted by ultrasonic, then taking out, centrifuging and standing to split the phases, calculating the quality of the alliin in the upper phase and the lower phase respectively, thereby obtaining the yield of the alliin in the upper phase and the lower phase. The method has high extraction yield of alliin with higher purity, has low cost, uses simple equipment, is simple in operation, is convenient and simple for postprocessing of the extractive, and is easy for amplification and continuous operation; micromolecular organic solvent has small interfacial tension and is beneficial for mass transfer of two phases.
Description
(1) technical field
The invention belongs to garlic intensive processing field, particularly a kind of ultrasonic auxiliary double water-phase extracts the method for alliin.
(2) background technology
Garlic is all widely used in treatment and prevention some diseases all the time, have been found that sulfocompound wherein has good medical effect, especially aspect preventing cancer, a kind of flavor precursors alliin (ALLIIN) wherein has prophylactic effect.Research shows that alliin has antiperoxide, anti-diabetic, the anti-oxidant effect such as antibacterial, antitumor and anticancer; Alliin is unstable, in the situation that raw garlic is crushed or shred, can there is enzymatic reaction and generate garlicin, the effects such as that garlicin also has is antibacterial, antitumor, Immunosuppression adjusting, but garlicin easily decomposes and is difficult for preserving, and alliin medicines and health protection characteristic and garlicin are more or less the same, and the stable easily preservation of alliin, so the best way is directly to obtain alliin.
From extracting alliin from garlic method, be mainly to adopt water or water-alcohol mixture directly to extract at present, but extraction time is long, and extraction yield is low.
Ultrasonic wave extraction is the strong cavitation effect, disturbance effect, the high acceleration that utilize Ultrasonic Radiation pressure to produce, smash and the multistage effect such as stirring action, increase molecular motion of material frequency and speed, increase solvent penetration power, thereby accelerate target component and enter solvent, promote the carrying out of extracting.
Aqueous two phase extraction technique is to utilize the difference that component distributes between two waters and the technology of carrying out the separating-purifying of component, aqueous two-phase extraction is because its content of water in system is high, single-stage separating-purifying efficiency is high, rate of mass transfer fast, the phase-splitting time is short, operational condition is gentle, required equipment is simple, process is easy to amplify and continuous operation, the features such as processing capacity is large, energy consumption is low, and find broad application.Double water-phase is successfully for extracting the fields such as protein purification, enzyme, microbiotic, metal and natural constituent.
The double-aqueous phase system that traditional polymer/polymer forms has good separation performance, but all more expensive for constructing the price of one-tenth gathering compound of double-aqueous phase system, for general biological product, separation costs is too high, from being irrational economically, and interfacial tension is large; Inorganic salt/lower alcohol cost is low, viscosity is little, and is easy to reclaim, so more and more receive an acclaim.
(3) summary of the invention
The present invention is in order to make up the deficiencies in the prior art, and a kind of simple to operate, method that ultrasonic auxiliary double water-phase that extraction yield is high extracts alliin is provided.
The present invention is achieved through the following technical solutions:
Ultrasonic auxiliary double water-phase extracts a method for alliin, comprises the steps:
(1) get inorganic salt, add pH be 2.0 Bloomsbury smooth-Robinson, Robert damping fluid dissolves, and adds organic solvent ethanol, n-propyl alcohol or Virahol, mixes standing phase-splitting, obtains double water-phase extraction agent;
(2) regulating the pH of double water-phase extraction agent is 2.0-4.0, in every 10g double water-phase extraction agent, add 0.1-0.3g garlic powder, adopt ultrasonic wave to carry out assisted extraction, ultrasonic power is 440-540W, extraction temperature is 20-40 ℃, extraction time is 20-40min, takes out standing phase-splitting after centrifugal 10min under 3000r/min after extraction;
(3) take out the lower phase of the alliin of writing a letter in reply, with the ethanolic soln precipitation of volume fraction 65-80%, concentrated, vacuum-drying, obtain alliin product.
The present invention adopts ultrasonic wave to assist lower alcohols/salt double-aqueous phase system (ATPS) to extract, the method is mixed lower alcohol and salt, utilize alliin and the impurity different solubility in water and organic phase, alliin is distributed in lower phase, and other most of Impurity Distribution is in upper phase, solid matter is distributed in two alternate interfaces, impurity can be removed together with solid, and and traditional method comparison, the extraction yield of this method significantly improves, extraction is completed with a separated step, simplified operation.
More excellent scheme of the present invention is:
In step (1), Bloomsbury is smooth-Robinson, Robert damping fluid by the phosphoric acid of 0.04mol/L, the acetic acid of the boric acid of 0.04mol/L, 0.04mol/L and the configuration of the sodium hydroxide of 0.2mol/L form.
In step (1), the total mass of double water-phase extraction agent is 10g, formulates 8 kinds of double-aqueous phase systems, and these 8 kinds of systems are 19% (w/w) (NH
4)
2sO
4/ 26% (w/w) ethanol, 15% (w/w) (NH
4)
2sO
4/ 26% (w/w) n-propyl alcohol, 15% (w/w) (NH
4)
2sO
4/ 25% (w/w) Virahol, 18% (w/w) ethanol/24% (w/w) NaH
2pO
4, 26% (w/w) n-propyl alcohol/20% (w/w) NaH
2pO
4, 26% (w/w) Virahol/20% (w/w) NaH
2pO
4, 26% (w/w) n-propyl alcohol/20% (w/w) Na
3c
6h
5o
7, 26% (w/w) Virahol/16% (w/w) Na
3c
6h
5o
7.
Selected (NH
4)
2sO
4/ n-propyl alcohol double-aqueous phase system is the optimum extraction agent of alliin, investigate the impact that ammonium sulfate concentrations is extracted alliin, the quality settings of extraction agent is 10g, extraction conditions is: system pH 2.0, ultrasonic power 540W, extracts 30 ℃ of temperature, extraction time 40min, n-propyl alcohol massfraction 20% (w/w), (NH
4)
2sO
4massfraction (w/w) 13%-25%.
Investigate the impact that n-propyl alcohol concentration is extracted alliin, the quality settings of extraction agent is 10g, and extraction conditions is: system pH 2.0g, ultrasonic power 540W, extracts 30 ℃ of temperature, extraction time 40min, n-propyl alcohol massfraction 17%-29% (w/w), (NH
4)
2sO
4massfraction (w/w) is 19%.
The impact that investigation system pH extracts alliin, the quality settings of extraction agent is 10g, extraction conditions is: system pH 2.0-10.0, ultrasonic power 540W, extracts 30 ℃ of temperature, extraction time 40min, n-propyl alcohol massfraction 20% (w/w), (NH
4)
2sO
4massfraction (w/w) is 19%.
Investigate the impact that ultrasonic power extracts alliin, the quality settings of extraction agent is 10g, and extraction conditions is: system pH 2.0, garlic powder 0.2g, (NH
4)
2sO
4massfraction (w/w) be 19%, the massfraction of n-propyl alcohol (w/w) is 20%, ultrasonic power 320-540W extracts 30 ℃ of temperature, extraction time 40min.
Investigate the impact that garlic opaque amount is extracted alliin, the quality settings of extraction agent is 10g, and extraction conditions is: system pH 2.0, and garlic opaque amount is 0.1,0.2,0.4,0.6,0.8,1.0g, (NH
4)
2sO
4massfraction (w/w) be 19%, the massfraction of n-propyl alcohol (w/w) is 20%, ultrasonic power 540W extracts 30 ℃ of temperature, extraction time 40min.
According to above monofactorial experimental result, to ultrasonic power, garlic opaque amount, system pH is optimized with response curved surface Box-Behnken design (BBD), the pH value of double water-phase extraction agent is set as 2.0,3.0 or 4.0, in every 10g double water-phase extraction agent, add 0.1g, 0.23g or 0.3g garlic powder, ultrasonic power is set as 440W, 500W or 540W; Preferably, double water-phase extraction agent is 19% (NH
4) SO
4/ 20% n-propyl alcohol double-aqueous phase system, pH is 2.35; In every 10g double water-phase extraction agent, add 0.23g garlic powder, ultrasonic power is set as 500W.
The present invention is high to the extraction yield of alliin, and purity is higher, adopts micromolecular organic solvent interfacial tension little, contributes to the mass transfer between two-phase, and cost is low, and equipment is simple, simple to operate, and extract aftertreatment is easy, is easy to amplify and operate continuously.
(4) accompanying drawing explanation
Below in conjunction with accompanying drawing, the present invention is further illustrated.
The affect schematic diagram of Fig. 1 ammonium sulfate concentrations on alliin partition ratio and extraction yield.
The affect schematic diagram of Fig. 2 n-propyl alcohol concentration on alliin partition ratio and extraction yield.
The affect schematic diagram of Fig. 3 pH value of solution on alliin partition ratio and extraction yield.
The affect schematic diagram of Fig. 4 garlic powder addition on alliin partition ratio and extraction yield.
The affect schematic diagram of Fig. 5 ultrasonic power on alliin partition ratio and extraction yield.
Fig. 6 Response Surface Method 3D figure pH and the affect schematic diagram of garlic powder addition on alliin yield;
Fig. 7 Response Surface Method 3D figure pH and the affect schematic diagram of ultrasonic power on alliin yield;
Fig. 8 Response Surface Method 3D figure garlic powder addition and the affect schematic diagram of ultrasonic power on alliin yield.
(5) embodiment
Embodiment 1:
(1) preparation double-aqueous phase system.Get massfraction and be 16% ammonium sulfate, add pH be 2.35 Bloomsbury smooth-Robinson, Robert damping fluid dissolves, and adds the n-propyl alcohol of 20% (w/w), is formulated as double water-phase extraction agent;
(2) ultrasonic auxiliary double water-phase extracts.In the double-aqueous phase system of step (1) gained, garlic powder (the source of garlic powder: cross 60 mesh sieves after garlic sheet is pulverized that adds 2% precision weighing, the garlic sheet using is the commercial product of routine, setting ultrasound condition is power 540W, 30 ℃ of temperature, time 40min, carrying out ultrasonic auxiliary double water-phase extracts, after extraction finishes, the centrifugal obvious alliin extracting solution of phase-splitting that obtains.
Measure the double water-phase volume of phase up and down, utilize Syrups by HPLC double water-phase up and down mutually in the concentration of alliin, by formula 3 calculating alliin extraction yields.
(1)，
(2)；
（3）。
In formula: R is for comparing;
v t with
v b lower is the volume of phase (ml) up and down;
c t ,
c b lower is respectively the upper and lower mass concentration (mg/mL) of middle alliin mutually; K is partition ratio;
m d for the garlic opaque amount adding.
The extraction yield of alliin is 20.1mg/g.
Embodiment 2:
(1) to get massfraction be 19% ammonium sulfate to preparation double-aqueous phase system, add pH be 2.35 Bloomsbury smooth-Robinson, Robert damping fluid dissolves, and adds 23% n-propyl alcohol, is formulated as double water-phase extraction agent;
(2) ultrasonic auxiliary double water-phase is carried in the double-aqueous phase system of orientation step (1) gained, garlic powder (the source of garlic powder: cross 60 mesh sieves after garlic sheet is pulverized that adds 2% (w/w), the garlic sheet using is the commercial product of routine), setting ultrasound condition is power 540W, 30 ℃ of temperature of extraction, extraction time 40min, carrying out ultrasonic auxiliary double water-phase extracts, after extraction finishes, the centrifugal obvious alliin extracting solution of phase-splitting that obtains.The extraction yield of gained alliin is 20.6mg/g.
Embodiment 3
(1) to get massfraction be 28% ammonium sulfate to preparation double-aqueous phase system, add pH be 2.35 Bloomsbury smooth-Robinson, Robert damping fluid dissolves, and then adds 17% n-propyl alcohol, is formulated as double water-phase extraction agent;
(2) ultrasonic auxiliary double water-phase is carried in the double-aqueous phase system of orientation step (1) gained, garlic powder (the source of garlic powder: cross 60 mesh sieves after garlic sheet is pulverized that adds 2% (w/w), the garlic sheet using is the commercial product of routine), setting ultrasound condition is power 540W, 30 ℃ of temperature of extraction, extraction time 40min, carrying out ultrasonic auxiliary double water-phase extracts, after extraction finishes, the centrifugal obvious alliin extracting solution of phase-splitting that obtains.The extraction yield of gained alliin is 19.2mg/g.
Table 1 extracts the selection of the different double-aqueous phase systems of alliin
Claims (5)
1. ultrasonic auxiliary double water-phase extracts a method for alliin, it is characterized by, and comprises the steps: that (1) get inorganic salt, add pH be 2.0 Bloomsbury smooth-Robinson, Robert damping fluid dissolves, add organic solvent ethanol, n-propyl alcohol or Virahol, mix standing phase-splitting, obtain double water-phase extraction agent; (2) regulating the pH of double water-phase extraction agent is 2.0-4.0, in every 10g double water-phase extraction agent, add 0.1-0.3g garlic powder, adopt ultrasonic wave to carry out assisted extraction, ultrasonic power is 440-540W, extraction temperature is 20-40 ℃, extraction time is 20-40min, takes out standing phase-splitting after centrifugal 10min under 3000r/min after extraction, obtains the obvious alliin extracting solution of phase-splitting; (3) take out the lower phase that is rich in alliin, by the ethanol precipitation of 65%-80%, concentrated, vacuum-drying, obtains alliin product.
2. ultrasonic auxiliary double water-phase according to claim 1 extracts the method for alliin, it is characterized in that: in step (1), Bloomsbury is smooth-Robinson, Robert damping fluid by the phosphoric acid of 0.04mol/L, the acetic acid of the boric acid of 0.04mol/L, 0.04mol/L and the configuration of the sodium hydroxide of 0.2mol/L form.
3. ultrasonic auxiliary double water-phase according to claim 1 extracts the method for alliin, it is characterized in that: in step (1), the total mass of double water-phase extraction agent is 10g, and be 16-28% (NH
4) SO
4/ 14-26% n-propyl alcohol double-aqueous phase system.
4. ultrasonic auxiliary double water-phase according to claim 3 extracts the method for alliin, it is characterized in that: in step (2), the pH value of double water-phase extraction agent is set as 2.35, and in every 10g double water-phase extraction agent, add 0.23g garlic powder, ultrasonic power is set as 500W.
5. ultrasonic auxiliary double water-phase according to claim 3 extracts the method for alliin, it is characterized in that: in step (2), double water-phase extraction agent is 19% (NH
4) SO
4/ 20% n-propyl alcohol double-aqueous phase system.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105399650A (en) * | 2015-12-16 | 2016-03-16 | 齐鲁工业大学 | Method for extracting alliin through reverse micelles system |
| CN109222005A (en) * | 2018-11-08 | 2019-01-18 | 育生（厦门）生物科技有限公司 | Improve the two stages processing method of S- allyl cysteine acid content in garlic |
| CN113512543A (en) * | 2021-06-08 | 2021-10-19 | 新疆农业职业技术学院 | Ultrasonic-based garlic allinase rapid extraction process |
| CN114478331A (en) * | 2022-02-17 | 2022-05-13 | 齐鲁工业大学 | Method for separating and purifying alliin |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110668982B (en) * | 2019-11-14 | 2021-03-30 | 齐鲁工业大学 | Preparation method of high-purity alliin |
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| Publication number | Priority date | Publication date | Assignee | Title |
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