CN104069157A - Method for composite enzyme-coordinated dual-frequency ultrasonic extraction of licoflavone - Google Patents
Method for composite enzyme-coordinated dual-frequency ultrasonic extraction of licoflavone Download PDFInfo
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- CN104069157A CN104069157A CN201410266464.7A CN201410266464A CN104069157A CN 104069157 A CN104069157 A CN 104069157A CN 201410266464 A CN201410266464 A CN 201410266464A CN 104069157 A CN104069157 A CN 104069157A
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Abstract
The invention discloses a method for extracting licoflavone from glycyrrhiza waste residues, and particularly relates to a method for composite enzyme-coordinated dual-frequency ultrasonic extraction of licoflavone. The method comprises the following steps: adding 8-10fold of 70-80% ethanol solution to glycyrrhiza waste residues, adding composite enzyme which is 0.5-2% of weight of the raw material, evenly agitating, and putting into a dual-frequency ultrasonic extractor to extract, so as to obtain enzymatic hydrolysate; inactivating at high temperature, centrifuging, taking supernate, and extracting with petroleum ether; taking an ethanol layer and concentrating into 1/10 of original volume; feeding to a pre-treated macroporous absorptive resin column, eluting by 5-7BV of 80-90% ethanol, and collecting an eluent and concentrating; and recrystallizing an ethanol solution containing 1-10% hydrochloric acid, so as to obtain the licoflavone. The glycyrrhiza waste residues are taken as the raw materials, and the target of reasonable development and utilization of resources is achieved. The method is simple in process, easily available in raw materials, high in yield, and applicable to large-scale industrial production.
Description
Technical field
The present invention relates to a kind of method of extracting licoflavone from licorice slag, be specifically related to the method that the collaborative double-frequency ultrasound of a kind of compound enzyme extracts licoflavone.
Background technology
Licoflavone is one of most important active component in Chinese crude drug Radix Glycyrrhizae, has the effects such as antiulcer, antibacterial, antiinflammatory, spasmolytic, blood fat reducing, analgesia, has also found that licoflavone has the effect of the acquired immune deficiency syndrome (AIDS) of preventing and treating since 1986.Up to now, from Radix Glycyrrhizae, isolated more than 150 flavone compound, they belong to respectively the tens large classes such as flavonoid, flavonols, osajin, chalcones, flavanone, dihydrochalcone-type.The research of licoflavone is more and more attracted much attention and paid attention to, can predict, licoflavone has broad application prospects.And China Radix Glycyrrhizae is the gesture of minimizing year by year, the market demand faces " running out of rice and fuel " predicament.For a long time, suitability for industrialized production is only extracted the glycyrrhizic acid in Radix Glycyrrhizae as main effective ingredient, and the abandoned of licorice slag after extracting, has caused environmental pollution and the wasting of resources.In licorice slag, contain and have in a large number the flavone compound of medical value and economic worth and a small amount of glycyrrhizic acid etc.Therefore, from licorice slag, extract licoflavone, not only can preserve the ecological environment, can also obtain economic benefit, kill two birds with one stone.
The extraction separation method that is usually used at present licoflavone has following several: water extraction method, ethanol/water solvent extraction method, alkali extraction and acid precipitation, macroporous resin adsorption partition method, supercritical fluid extraction etc.Its weak point is, solvent extraction method cost is high, and the cycle is long; Alkali extraction and acid precipitation can only extract part licoflavone; Supercritical extraction method equipment investment is large, is difficult to suitability for industrialized production.
Summary of the invention
The object of the invention is, a kind of method of extracting licoflavone from licorice slag is provided, the licoflavone extracting by the method, extraction ratio is high, and purity can reach more than 80%, can realize suitability for industrialized production.
Technical scheme of the present invention is carried out according to the following step:
Licorice slag is added to 70 ~ 80% the alcoholic solution of 8 ~ 10 times, the compound enzyme (cellulase: pectase: protease is 0.4:0.4:0.2) that adds again raw material weight 0.5 ~ 2%, stir, be placed in double-frequency ultrasound extractor and extract 10 ~ 30min, supersonic frequency is respectively 25Hz, 40Hz, ultrasonic power is 300 ~ 400W, enzymolysis solution is in 80 ℃ of high-temperature inactivation 15min, centrifugal, get supernatant, petroleum ether extraction, get alcohol layer and be concentrated into 1/10 of original volume, the pretreated macroporous adsorptive resins of upper process, 80 ~ 90% the ethanol elution of 5 ~ 7BV, collection eluent is concentrated, hydrochloric 1 ~ 10% alcoholic solution recrystallization 2 times, obtain licoflavone.
Ultrasonic power of the present invention is that double-frequency ultrasound is made used time general power (25KHz and 40KHz respectively get half).
The preprocess method of macroporous adsorbent resin of the present invention is for first fully soaking resin with ethanol, then with ethanol, be washed till effluent and add suitable quantity of water without white opacity, with deionized water, be washed till without alcohol taste again, use 5% HCl solution soaking 3h instead, it is neutral with deionized water, being washed till pH value again, follow the NaOH solution soaking 3h with 5%, it is neutral finally with deionized water, being washed till pH value.
Macroreticular resin of the present invention is HP-20, D-101 or NKA-9.
Tool of the present invention has the following advantages:
(1) the present invention be take licorice slag as raw material, is conducive to alleviate the crisis of existing Licorice scarcity, preserves the ecological environment, and has good Social benefit and economic benefit;
(2) the present invention utilizes the collaborative double-frequency ultrasound extraction of compound enzyme licoflavone method can reach following effect: extraction time is short, and extraction ratio is high, and energy resource consumption is low;
(3) adopt petroleum ether extraction, can remove the oil-soluble impurities in licorice slag;
(4) through these pretreatment, upper prop liquid has effectively been removed impurity, and the pollution to resin after upper prop is little, and product yield is high, and purity is large, is suitable for large-scale production.
Below in conjunction with the specific embodiment, further illustrate the present invention, but the scope of protection of present invention is not limited to following embodiment.
The specific embodiment
Example 1:
1kg licorice slag is added to 80% the alcoholic solution of 8 times, the compound enzyme (cellulase: pectase: protease is 0.4:0.4:0.2) that adds again raw material weight 0.5%, stir, be placed in double-frequency ultrasound extractor and extract 30min, supersonic frequency is respectively 25Hz, 40Hz, ultrasonic power is 300W, enzymolysis solution is in 80 ℃ of high-temperature inactivation 15min, centrifugal, get supernatant, petroleum ether extraction, get alcohol layer and be concentrated into 1/10 of original volume, the pretreated D-101 macroporous adsorptive resins of upper process, 80% the ethanol elution of 5BV, collection eluent is concentrated, hydrochloric 1% alcoholic solution recrystallization 2 times, obtain licoflavone 30g, through UV, detect, purity is 85.4%.
Example 2:
1kg licorice slag is added to 75% the alcoholic solution of 9 times, the compound enzyme (cellulase: pectase: protease is 0.4:0.4:0.2) that adds again raw material weight 1.5%, stir, be placed in double-frequency ultrasound extractor and extract 20min, supersonic frequency is respectively 25Hz, 40Hz, ultrasonic power is 400W, enzymolysis solution is in 80 ℃ of high-temperature inactivation 15min, centrifugal, get supernatant, petroleum ether extraction, get alcohol layer and be concentrated into 1/10 of original volume, the pretreated HP-20 macroporous adsorptive resins of upper process, 90% the ethanol elution of 7BV, collection eluent is concentrated, hydrochloric 5% alcoholic solution recrystallization 2 times, obtain licoflavone 29g, through UV, detect, purity is 86.2%.
Example 3:
1kg licorice slag is added to 70% the alcoholic solution of 10 times, the compound enzyme (cellulase: pectase: protease is 0.4:0.4:0.2) that adds again raw material weight 2%, stir, be placed in double-frequency ultrasound extractor and extract 10min, supersonic frequency is respectively 25Hz, 40Hz, ultrasonic power is 350W, enzymolysis solution is in 80 ℃ of high-temperature inactivation 15min, centrifugal, get supernatant, petroleum ether extraction, get alcohol layer and be concentrated into 1/10 of original volume, the pretreated NKA-9 macroporous adsorptive resins of upper process, 85% the ethanol elution of 6BV, collection eluent is concentrated, hydrochloric 10% alcoholic solution recrystallization 2 times, obtain licoflavone 32g, through UV, detect, purity is 84.1%.
Claims (6)
1. a compound enzyme is worked in coordination with the method that double-frequency ultrasound extracts licoflavone, it is characterized in that: licorice slag is added to 70 ~ 80% the alcoholic solution of 8 ~ 10 times, the compound enzyme that adds again raw material weight 0.5 ~ 2%, stir, be placed in double-frequency ultrasound extractor and extract 10 ~ 30min, supersonic frequency is respectively 25Hz, 40Hz, ultrasonic power is 300 ~ 400W, enzymolysis solution is in 80 ℃ of high-temperature inactivation 15min, centrifugal, get supernatant, petroleum ether extraction, get alcohol layer and be concentrated into 1/10 of original volume, the pretreated macroporous adsorptive resins of upper process, 80 ~ 90% the ethanol elution of 5 ~ 7BV, collection eluent is concentrated, hydrochloric 1 ~ 10% alcoholic solution recrystallization 2 times, obtain licoflavone.
2. the collaborative double-frequency ultrasound of a kind of compound enzyme according to claim 1 extracts the method for licoflavone, it is characterized in that, described compound enzyme is cellulase, pectase and protease.
3. the collaborative double-frequency ultrasound of a kind of compound enzyme according to claim 1 and 2 extracts the method for licoflavone, it is characterized in that, the weight ratio of described cellulase, pectase and protease is 0.4:0.4:0.2.
4. the collaborative double-frequency ultrasound of a kind of compound enzyme according to claim 1 extracts the method for licoflavone, it is characterized in that, described ultrasonic power is that double-frequency ultrasound is made used time general power (25KHz and 40KHz respectively get half).
5. the collaborative double-frequency ultrasound of a kind of compound enzyme according to claim 1 extracts the method for licoflavone, it is characterized in that, the preprocess method of described macroporous adsorbent resin is for first fully soaking resin with ethanol, then with ethanol, be washed till effluent and add suitable quantity of water without white opacity, with deionized water, be washed till without alcohol taste again, use 5% HCl solution soaking 3h instead, then with deionized water, to be washed till pH value be neutral, follow the NaOH solution soaking 3h with 5%, it is neutral finally with deionized water, being washed till pH value.
6. the collaborative double-frequency ultrasound of a kind of compound enzyme according to claim 1 extracts the method for licoflavone, it is characterized in that, described macroreticular resin is HP-20, D-101 or NKA-9.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107012185A (en) * | 2016-01-28 | 2017-08-04 | 亿利资源集团有限公司 | The preparation method and composite microporous separator of radix glycyrrhizae cellulose |
| CN115260024A (en) * | 2022-08-12 | 2022-11-01 | 守正创新生物科技(天津)有限公司 | Method for preparing high-purity licochalcone A from glycyrrhiza inflata |
-
2014
- 2014-06-16 CN CN201410266464.7A patent/CN104069157A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107012185A (en) * | 2016-01-28 | 2017-08-04 | 亿利资源集团有限公司 | The preparation method and composite microporous separator of radix glycyrrhizae cellulose |
| CN115260024A (en) * | 2022-08-12 | 2022-11-01 | 守正创新生物科技(天津)有限公司 | Method for preparing high-purity licochalcone A from glycyrrhiza inflata |
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