CN112724155B - Method for preparing white glabridin by subcritical technology - Google Patents
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Abstract
A method for preparing white glabridin by subcritical technique, relate to the preparation field of glabridin, the invention extracts white glabridin from Glycyrrhiza glabra Linne, the extraction process is simple and convenient, the operability is strong; the yield of effective substances is high, and the prepared low-content glabridin has better color, lower processing cost and the like compared with the current mainstream glabridin in the market while fully utilizing resources, and is suitable for large-scale popularization and application.
Description
Technical Field
The invention relates to the field of glabridin preparation, in particular to a method for preparing white glabridin by adopting a subcritical technology.
Background
It is known that glabridin is a flavonoid substance extracted from a precious plant called glycyrrhiza glabra, which is reputed as "whitening gold" because of its powerful whitening and high cost effects, can eliminate free radicals and basal melanin, and is a whitening and anti-aging cherry for skin.
Glabridin is one of the main flavonoid components in glycyrrhiza glabra. It has strong anti-free radical oxidation effect in cytochrome P450/NADPH oxidation system, and can obviously inhibit free radicals generated in vivo metabolism process, so as to prevent biomacromolecule (low density lipoprotein LDL, DNA) sensitive to oxidation and cell wall and the like from being damaged by free radical oxidation. Thereby preventing and treating certain pathological changes related to free radical oxidation, such as atherosclerosis, cell aging and the like. In addition, glabridin has certain functions of lowering blood fat and lowering blood pressure. Italian studies have also demonstrated that glabridin has an appetite suppressant effect, which reduces fat without losing weight. Through repeated tests, the medicine mainly has the following effects:
1. whitening and inhibiting melanogenesis;
2. anti-inflammatory effects;
3. antioxidation;
4. antibacterial, and the like.
Since glabridin is a flavone compound extracted from Glycyrrhiza glabra Linne, it has whitening, antiinflammatory, and ultraviolet resisting effects. The fixed specification of glabridin in the market is mainly 40% and 90% in content, 40% of glabridin products are yellow and affect the color of cosmetics after being added into the cosmetics, and 90% of glabridin products are white, but the production cost is high, so that the glabridin cannot be widely applied to the cosmetics.
How to provide a method for preparing white glabridin by adopting a subcritical technology becomes a long-term technical demand of the technical personnel in the field.
Disclosure of Invention
In order to overcome the defects in the background technology, the invention provides a method for preparing white glabridin by adopting a subcritical technology, the white glabridin is extracted from glycyrrhiza glabra, the extraction process is simple and convenient, the operability is strong, the color is better, the processing cost is lower, and the like, and the market requirements are completely met.
In order to achieve the above purpose, the invention adopts the following technical scheme:
a method for preparing white glabridin by adopting a subcritical technology comprises the following steps:
firstly, adding ammonia water or any one of sodium carbonate or sodium hydroxide into pure water at normal temperature to adjust the pH value to 9-9.5 to obtain an aqueous solution, then extracting the crushed glycyrrhiza glabra in the aqueous solution for 3-4 hours, wherein the material-liquid ratio of the crushed glycyrrhiza glabra to the aqueous solution is 1: 10: 8-1, separating grass residue from an extracting solution, reserving filtrate, repeatedly extracting for 4-5 times, and drying the extracted grass residue for later use;
secondly, placing the grass residue obtained in the previous step into a high-pressure extraction kettle, extracting by using subcritical fluid 1,1,1, 2-tetrafluoroethane (R134a), and collecting an extraction product under the process conditions of extraction pressure of 10 MPa-12 MPa, extraction temperature of 45-50 ℃ and extraction time of 80-90 min;
dissolving the extraction product obtained in the previous step in 90-99% methanol or ethanol, adding water to dilute the solution to 40-60% alcohol concentration, placing the solution in a refrigerator at 0-5 ℃ for standing for 2-6 hours, and separating supernatant;
fourthly, taking the supernatant collected in the previous step, decoloring the supernatant by silica gel decoloring sand which accounts for 10-20% of the dry matter, keeping the temperature at 30-40 ℃ for 0.5-2 hours, filtering and separating the silica gel decoloring sand, and reserving the supernatant for later use;
fifthly, purifying the supernatant obtained in the previous step by using decolorizing resin, wherein the volume of the decolorizing resin is 10-15 times of the weight of dry matter of the supernatant, the sample loading speed is 1-2 BV/h, and collecting the sample to flow out;
and sixthly, concentrating and drying the sample loading outflow obtained in the previous step into powder to obtain the white glabridin with the content of 30-40%.
In the method for preparing white glabridin by using the subcritical technology, the pH value of the aqueous solution in the first step is preferably 9.2.
In the method for preparing white glabridin by using the subcritical technology, the extraction pressure in the second step is preferably 11 MPa.
In the method for preparing white glabridin by using the subcritical technology, the extraction temperature in the second step is preferably 47 ℃.
In the method for preparing white glabridin by using the subcritical technology, the concentration of the dissolved methanol in the third step is preferably 99%.
In the method for preparing white glabridin by using the subcritical technology, the refrigerator temperature in the third step is preferably 3 ℃.
In the method for preparing white glabridin by using the subcritical technology, the decoloring temperature in the fourth step is preferably 35 ℃.
In the method for preparing white glabridin by using subcritical technology, the type of the decolorizing resin in the fifth step is any one of LX-94, LX-98, LX-T5, LS-700B or LS-807A.
In the method for preparing white glabridin by using subcritical technology, the model of the decolorizing resin is preferably LX-94.
In the method for preparing white glabridin by using the subcritical technology, the volume of the decolorizing resin in the fifth step is preferably 12 times of the weight of the supernatant dry matter.
By adopting the technical scheme, the invention has the following advantages:
the white glabridin is extracted from the glycyrrhiza glabra, the extraction process is simple and convenient, and the operability is strong; the yield of effective substances is high, and the prepared low-content glabridin has better color, lower processing cost and the like compared with the current mainstream glabridin in the market while fully utilizing resources, and is suitable for large-scale popularization and application.
Detailed Description
The present invention will be explained in more detail by the following examples, which are not intended to limit the invention;
the invention relates to a method for preparing white glabridin by adopting a subcritical technology, which specifically comprises the following steps:
firstly, adding ammonia water or any one of sodium carbonate or sodium hydroxide into pure water at normal temperature to adjust the pH value to 9-9.5 to obtain an aqueous solution, wherein the pH value of the aqueous solution is preferably 9.2, then extracting the crushed glycyrrhiza glabra in the aqueous solution for 3-4 hours, and the material-to-liquid ratio of the crushed glycyrrhiza glabra to the aqueous solution is 1: 10: 8-1, separating grass residue from an extracting solution, reserving filtrate, repeatedly extracting for 4-5 times, and drying the extracted grass residue for later use;
secondly, placing the grass residue obtained in the previous step into a high-pressure extraction kettle, extracting by using subcritical fluid 1,1,1, 2-tetrafluoroethane (R134a), and collecting an extraction product under the process conditions of extraction pressure of 10 MPa-12 MPa, extraction temperature of 45-50 ℃ and extraction time of 80-90 min; in specific implementation, the extraction pressure is preferably 11MPa, and the extraction temperature is preferably 47 ℃;
dissolving the extraction product obtained in the previous step in 90-99% methanol or ethanol, adding water to dilute the solution to 40-60% alcohol concentration, placing the solution in a refrigerator at 0-5 ℃ for standing for 2-6 hours, and separating supernatant; in specific implementation, water is added for dilution until the concentration of 50% alcohol is optimal, standing in a refrigerator for 4 hours is optimal, the concentration of dissolved methanol is preferably 99%, and the temperature of the refrigerator is preferably 3 ℃;
fourthly, taking the supernatant collected in the previous step, decoloring the supernatant by silica gel decoloring sand which accounts for 10-20% of the dry matter, keeping the temperature at 30-40 ℃ for 0.5-2 hours, filtering and separating the silica gel decoloring sand, and reserving the supernatant for later use; in specific implementation, the decolorizing temperature is preferably 35 ℃;
fifthly, purifying the supernatant obtained in the previous step by using decolorizing resin, wherein the volume of the decolorizing resin is 10-15 times of the weight of dry matter of the supernatant, the sample loading speed is 1-2 BV/h, and collecting the sample to flow out; in specific implementation, the model of the decolorizing resin is any one of LX-94, LX-98, LX-T5, LS-700B or LS-807A, and the model of the decolorizing resin is preferably LX-94; the volume of the decolorizing resin is 12 times of the weight of the dry matter of the supernatant, and the loading speed is preferably 1.5 BV/h;
and sixthly, concentrating and drying the sample loading outflow obtained in the previous step into powder to obtain the white glabridin with the content of 30-40%.
The specific embodiment of the invention is as follows:
example one:
firstly, adding ammonia water into pure water at normal temperature to adjust the pH value to 9.2 to obtain an aqueous solution, then extracting the crushed glycyrrhiza glabra in the aqueous solution for 4 hours, wherein the ratio of the crushed glycyrrhiza glabra to the aqueous solution is 1: 8, separating grass residue from an extracting solution, reserving filtrate, repeatedly extracting for 4 times, and then drying the extracted grass residue for later use;
secondly, placing the grass residues in a high-pressure extraction kettle, extracting by using subcritical fluid 1,1,1, 2-tetrafluoroethane (R134a), and collecting an extraction product under the process conditions of extraction pressure of 10MPa, extraction temperature of 50 ℃ and extraction time of 90 min;
thirdly, dissolving the extraction product obtained in the previous step in 90-99% methanol or ethanol, adding water to dilute the solution to 50% methanol concentration, placing the solution in a refrigerator at 0-5 ℃ for standing for 4 hours, and separating supernatant;
fourthly, taking the supernatant collected in the previous step, decoloring the supernatant by using silica gel decoloring sand with the mass of 20 percent of the dry mass, keeping the temperature at 40 ℃ for 2 hours, filtering and separating the silica gel decoloring sand, and reserving the supernatant for later use;
fifthly, purifying the supernatant solution obtained in the previous step by using decolorizing resin LX-94, wherein the volume of the resin is 15 times of the weight of dry matter of the supernatant, the sample loading speed is 1BV/h, and collecting the sample to flow out;
and sixthly, concentrating and drying the sample loading outflow obtained in the previous step into powder to obtain the white glabridin with the content of 30-40%.
Example two:
firstly, adding ammonia water into pure water at normal temperature to adjust the pH value to 9 to obtain an aqueous solution, then extracting the crushed glycyrrhiza glabra in the aqueous solution for 4 hours, wherein the ratio of the crushed glycyrrhiza glabra to the aqueous solution is 1: 9, separating grass residue from the extracting solution, reserving the filtrate, repeatedly extracting for 5 times, and then drying the extracted grass residue for later use;
secondly, placing the grass residues in a high-pressure extraction kettle, extracting by using subcritical fluid 1,1,1, 2-tetrafluoroethane (R134a), and collecting an extraction product under the process conditions of extraction pressure of 12MPa, extraction temperature of 40 ℃ and extraction time of 80 min;
thirdly, dissolving the extraction product obtained in the last step in 99% methanol, then adding water to dilute the solution to 50% methanol concentration, placing the solution in a refrigerator at 0-5 ℃ for standing for 4 hours, and separating supernatant;
fourthly, taking the supernatant collected in the previous step, decoloring the supernatant by silica gel decoloring sand with the mass of 15 percent of the dry mass, keeping the temperature at 35 ℃ for 1.5 hours, filtering and separating the silica gel decoloring sand, and reserving the supernatant for later use;
fifthly, purifying the supernatant solution obtained in the previous step by using decolorizing resin LX-T5, wherein the volume of the resin is 12 times of the weight of dry matter of the supernatant, the sample loading speed is 1.2BV/h, and collecting the sample loading outflow;
and sixthly, concentrating and drying the sample obtained in the previous step to obtain powder, namely the white glabridin with the content of 30-40%.
Example three:
firstly, adding ammonia water into pure water at normal temperature to adjust the pH value to 9.2 to obtain an aqueous solution, then extracting the crushed glycyrrhiza glabra in the aqueous solution for 4 hours, wherein the ratio of the crushed glycyrrhiza glabra to the aqueous solution is 1:10, separating grass residue from an extracting solution, reserving filtrate, repeatedly extracting for 5 times, and then drying the extracted grass residue for later use;
secondly, placing the grass residues in a high-pressure extraction kettle, extracting by using subcritical fluid 1,1,1, 2-tetrafluoroethane (R134a), and collecting an extraction product under the process conditions of extraction pressure of 10MPa, extraction temperature of 30 ℃ and extraction time of 90 min;
thirdly, dissolving the extraction product obtained in the previous step in 95% methanol, adding water to dilute the solution to 50% methanol concentration, standing the solution in a refrigerator at 0-5 ℃ for 4 hours, and separating supernatant;
fourthly, taking the supernatant collected in the previous step, decoloring the supernatant by silica gel decoloring sand with the mass of 15 percent of the dry mass, keeping the temperature at 35 ℃ for 2 hours, filtering and separating the silica gel decoloring sand, and reserving the supernatant for later use;
fifthly, purifying the supernatant solution obtained in the previous step by using decolorizing resin LS-807A, wherein the volume of the resin is 10 times of the weight of the dry matter of the supernatant, the sample loading speed is 1.5BV/h, and collecting the sample to flow out;
and sixthly, concentrating and drying the sample loading outflow obtained in the previous step into powder to obtain the white glabridin with the content of 30-40%.
Examples of applications of the invention are as follows:
the main active ingredient of the licorice root extract is glabridin, and the concentration of the licorice root extract used in the skin whitening agent is 10-40%. Glabridin has been shown to be 16 times more depigmenting than hydroquinone. The research on the effect time of the glabridin whitening effect also finds that the glabridin has quick effect. The whitening effect is very obvious after seven days. Clinical comparative tests of whitening skin by 0.4 percent of liquorice extract, 0.05 percent of betamethasone and 0.05 percent of retinoic acid, which are completed in Thailand, show that 70 percent of patients think that the effect of the liquorice extract is excellent.
The whitening effect evaluation of the whitening cream with glabridin as a main component is carried out by a company in Guangdong by adopting a 3D melanin skin model in-vitro test and a human body test method. The in vitro test result of the 3D melanin skin model shows that the whitening cream has a remarkable inhibiting effect on the synthesis or the transportation of melanin and has an excellent in vitro whitening effect. The results of human body tests show that after the whitening cream is used for 4 weeks, the melanin content (MI value) of the skin of a subject is obviously reduced, and the whiteness (L value) and the brightness (ITA DEG value) of the skin are increased, which shows that the whitening cream has good whitening effect.
The invention has the following beneficial effects:
1. the low content of white glabridin meets the market demand;
2. the process is simple, and the operability is strong;
3. high yield of effective substances, full utilization of resources and the like.
The present invention is not described in detail in the prior art.
The embodiments selected for the purpose of disclosing the invention, are presently considered to be suitable, it being understood, however, that the invention is intended to cover all variations and modifications of the embodiments which fall within the spirit and scope of the invention.
Claims (9)
1. A method for preparing white glabridin by adopting a subcritical technology is characterized by comprising the following steps: the method specifically comprises the following steps:
firstly, adding ammonia water or any one of sodium carbonate or sodium hydroxide into pure water at normal temperature, adjusting the p H value to 9-9.5 to obtain an aqueous solution, extracting the crushed glycyrrhiza glabra in the aqueous solution for 3-4 hours, wherein the material-liquid ratio of the crushed glycyrrhiza glabra to the aqueous solution is 1: 10: 8-1, separating grass residue from an extracting solution, reserving filtrate, repeatedly extracting for 4-5 times, and drying the extracted grass residue for later use;
secondly, placing the grass residue obtained in the previous step into a high-pressure extraction kettle, extracting by using subcritical fluid 1,1,1, 2-tetrafluoroethane (R134a), and collecting an extraction product under the process conditions of extraction pressure of 10 MPa-12 MPa, extraction temperature of 45-50 ℃ and extraction time of 80-90 min;
dissolving the extraction product obtained in the previous step in 90-99% methanol or ethanol, adding water to dilute the solution to 40-60% alcohol concentration, placing the solution in a refrigerator at 0-5 ℃ for standing for 2-6 hours, and separating supernatant;
fourthly, taking the supernatant collected in the previous step, decoloring the supernatant by silica gel decoloring sand which accounts for 10-20% of the dry matter, keeping the temperature at 30-40 ℃ for 0.5-2 hours, filtering and separating the silica gel decoloring sand, and reserving the supernatant for later use;
fifthly, purifying the supernatant obtained in the previous step by using decolorizing resin, wherein the volume of the decolorizing resin is 10-15 times of the weight of dry matter of the supernatant, the sample loading speed is 1-2 BV/h, and collecting the sample to flow out; the model of the decolorizing resin is any one of LX-94, LX-98, LX-T5 or LS-807A;
and sixthly, concentrating and drying the sample loading outflow obtained in the previous step into powder to obtain the white glabridin with the content of 30-40%.
2. The method for preparing white glabridin by subcritical technique according to claim 1, wherein: the aqueous solution in the first step had an p H value of 9.2.
3. The method for preparing white glabridin by subcritical technique according to claim 1, wherein: the extraction pressure in the second step is 11 MPa.
4. The method for preparing white glabridin by subcritical technique according to claim 1, wherein: the extraction temperature in the second step was 47 ℃.
5. The method for preparing white glabridin by subcritical technique according to claim 1, wherein: the concentration of the dissolved methanol in the third step was 99%.
6. The method for preparing white glabridin by subcritical technique according to claim 1, wherein: the temperature of the refrigerator in the third step was 3 ℃.
7. The method for preparing white glabridin by subcritical technique according to claim 1, wherein: the decoloring temperature in the fourth step is 35 ℃.
8. The method for preparing white glabridin by subcritical technique according to claim 1, wherein: the model of the decolorizing resin is LX-94.
9. The method for preparing white glabridin by subcritical technique according to claim 1, wherein: the volume of the decolorized resin in the fifth step is preferably 12 times the weight of the supernatant dry matter.
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| JP3827593B2 (en) * | 2002-03-14 | 2006-09-27 | 株式会社ノエビア | Topical skin preparation |
| CN101735233B (en) * | 2009-12-11 | 2011-11-16 | 暨南大学 | Method for producing high-purity glabridin |
| CN102250107A (en) * | 2011-06-07 | 2011-11-23 | 南京泽朗农业发展有限公司 | Method for preparing glabridin |
| CN104277048B (en) * | 2013-07-10 | 2016-09-21 | 岳普湖天瑞生物工程有限公司 | A kind of preparation method of Glycyrrhiza glabra L. total flavones |
| CN103360404B (en) * | 2013-07-30 | 2015-10-14 | 天津尚美化妆品有限公司 | A kind of method extracting high-purity glabrene from glycyrrhiza glabra |
| CN104861015A (en) * | 2015-01-16 | 2015-08-26 | 李玉山 | Synergistic clean extraction method of effective components in licorice root |
| CN104725394A (en) * | 2015-03-02 | 2015-06-24 | 李玉山 | Method for preparing glabridin |
| CN108176079B (en) * | 2017-12-29 | 2021-04-27 | 无限极（中国）有限公司 | A method for decolorizing Glycyrrhrizae radix extract |
| CN109126190A (en) * | 2018-08-27 | 2019-01-04 | 德西亚生物科技江苏有限公司 | A kind of extracting biologically active ingredients from subcritical fluid device and method |
| CN109081843A (en) * | 2018-08-30 | 2018-12-25 | 抚顺天润生物技术有限公司 | The preparation method of glabridin and the glabridin obtained by the preparation method, cosmetics |
| CN209361901U (en) * | 2019-01-03 | 2019-09-10 | 湖州欧利生物科技有限公司 | Environment-friendly type glabridin extraction element |
| CN109395427A (en) * | 2019-01-03 | 2019-03-01 | 湖州欧利生物科技有限公司 | A kind of environment-friendly type glabridin extraction element and its technique |
| CN110538112A (en) * | 2019-09-26 | 2019-12-06 | 无限极（中国）有限公司 | Glycyrrhiza glabra extract composition with function of enhancing skin immune barrier and preparation method and application thereof |
| CN110551137B (en) * | 2019-09-29 | 2022-06-10 | 浙江惠松制药有限公司 | Method for extracting and purifying glabridin and application of glabridin in cosmetics |
| CN110922413A (en) * | 2019-12-10 | 2020-03-27 | 辰风天然本草(北京)科技有限公司 | Extraction and separation method of glabridin |
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