CN103611333A - Method for supercritical extraction of super natural factors of propolis by using CO2 fluid in combination with solid-state enzymolysis - Google Patents
Method for supercritical extraction of super natural factors of propolis by using CO2 fluid in combination with solid-state enzymolysis Download PDFInfo
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- CN103611333A CN103611333A CN201310593710.5A CN201310593710A CN103611333A CN 103611333 A CN103611333 A CN 103611333A CN 201310593710 A CN201310593710 A CN 201310593710A CN 103611333 A CN103611333 A CN 103611333A
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Abstract
The invention provides a method for supercritical extraction of super natural factors of propolis by using CO2 fluid in combination with solid-state enzymolysis, and belongs to the technical field of propolis extraction and processing. The method comprises the following steps of: (1) raw material pretreatment: selecting good-quality raw propolis as the raw material, removing impurities, freezing and crushing; (2) solid-state enzymolysis of the raw material: adding an enzymolysis agent, homogenizing and mixing, and then filling the mixture on a solid-state enzymolysis bed, and carrying out solid-state enzymolysis by adjusting parameters such as enzymolysis temperature, humidity, pH and time; and (3) supercritical CO2 fluid extraction. The super natural factors of propolis obtained by the method provided by the invention are complete in components; in particular, the content of flavonoid aglycones such as myricetin, quercetin, apigenin and pinocembrin is increased; as a result, the antioxidant activity of the propolis extracts is enhanced; the propolis can be better applied to the fields such as medicines, health care food and cosmetics.
Description
Technical field
The present invention relates to the extracting process of active ingredient in a kind of propolis, particularly a kind of solid enzymatic hydrolysis-CO
2the method of the super natural factor of fluid supercritical extract propolis, belongs to propolis and extracts processing technique field.
Background technology
To be worker bee in bee colony mix with self glandula digestive and the glandular secretion thing (as beeswax and multiple digestive ferment) such as wax gland a kind of stickiness jelly forming afterwards by picking up from the young shoot of the plants such as willow, willow, pine tree and the resinoid of callus secretion to propolis (propolis).Propolis composition is very complicated, now separated and evaluation material out reaches kind more than 300, wherein flavonoids, terpenes, aliphatic category and aromatic acid and ester type compound thereof have obvious health care to human body, as antibacterial, anti-inflammatory, antiviral, hypoglycemic blood fat, enhancing immunological regulation and antitumor action etc.
Recently, by supercritical CO
2the technology of fluid extraction is applied to during propolis extracts, and particularly the extraction of, armaticity terpene rare class material strong to some the high attached value in propolis, heat sensitivity, the less material of lipophilic molecules amount has unique effect.Use supercritical CO
2the propolis that the technology extraction of fluid extraction obtains has the advantages such as color and luster is yellowish, no solvent residue, safety and Health, impurity is few, content of beary metal is low.But supercritical CO
2there are some technical barriers in fluid extraction propolis technology: as material, hardens, hooks stream; To too large as not thorough in the separating and extracting of the materials such as glycoside, polysaccharide of number molecular weight, thereby it is incomplete to cause extracting active ingredient, extraction yield is on the low side etc., these problems have restricted supercritical extraction technique applying in effective bee glue components extracts, and how breaking through these problems is directions that the present invention studies.
Solid enzymatic hydrolysis-CO
2the super natural factor technique of fluid supercritical extract propolis first carries out solid enzymatic hydrolysis by propolis raw material exactly, then adopts CO
2the super natural factor of fluid supercritical extraction technique bee glue extracting.Its basic subject is according to being the height selectivity of utilizing enzyme reaction, by the effect of enzyme, in propolis, some is difficult to by CO
2the polarity of fluid extraction is large, the material of molecular chain length carries out the degraded of molecular structure, become the little or short chain small-molecule substance of polarity, as the glycoside in propolis, polysaccharose substance are degraded into glucosides metaclass material by solid enzymatic hydrolysis technology, make that its active ingredient is free, suspendible or peptization in solute, then adopt CO
2the super natural factor of fluid supercritical extraction technique bee glue extracting, thus reach the object that improves its extraction efficiency.
Summary of the invention
Technical problem to be solved by this invention is solid enzymatic hydrolysis-CO
2the super natural factor of fluid supercritical extract propolis, object is further to improve supercritical CO
2fluid extraction technology level, improves extraction yield, reduces production costs, so that supercritical CO
2fluid extraction technology is widely applied in propolis extracts.The method technique is simple, easy to operate, and organic solvent-free has improved greatly production efficiency and can greatly improve the extraction content of flavone compound.
For solving the problems of the technologies described above, technical scheme of the present invention is:
A kind of solid enzymatic hydrolysis-CO
2the method of the super natural factor of fluid supercritical extract propolis, is characterized in that, its preparation method is as follows:
1, the pretreatment of hair glue: hair glue, through preliminary election classification, is removed impurity.Put into refrigerator, be placed in freezing 48h at-18 ℃ of temperature, the hair glue after freezing; put into rapidly disintegrating machine mode broken or employing liquid nitrogen frozen and carry out fragmentation, be broken into 10-20 object particle, the raw material after fragmentation; enter immediately 20 object reciprocating sieves, be greater than 20 object particles and return to refrigerator; ;
2, solid enzymatic hydrolysis: hair Jiao Peiyongshui and the complex enzyme system after crushing and screening is mixed, after mixed material is layered on solid enzymatic hydrolysis bed, regulate corresponding humiture, pH value, carry out solid enzymatic hydrolysis reaction;
3, CO
2fluid supercritical extract: the propolis grain packing through solid enzymatic hydrolysis, in extraction kettle, is injected to CO
2fluid extracts, and after extraction, system pressure is reduced to normal pressure, and extract is separated out, and obtains the super natural factor of propolis.
Above-mentioned a kind of solid enzymatic hydrolysis-CO
2the method of the super natural factor of fluid supercritical extract propolis, wherein, described solid enzymatic hydrolysis reaction enzymes is selected β-rhamnosidase, its enzyme dosage is 50-100IU/mg, beta-glucosidase, its enzyme dosage is 50-100IU/mg, zytase, its enzyme dosage is 1000-2000IU/mg, pectase, its enzyme dosage is 20-50IU/mg, AMS, its enzyme dosage is 10-50IU/mg or complex enzyme system, enzyme-added total amount is 0.5-1.5% by weight percentage, hydrolysis temperature 50-60 ℃, enzymolysis humidity 30-50%, enzymolysis pH3.5-5.0, enzymolysis time 12-24h.
Above-mentioned a kind of solid enzymatic hydrolysis-CO
2the method of the super natural factor of fluid supercritical extract propolis, wherein, described CO
2the process conditions of fluid supercritical extract are as follows: extracting pressure 25-35MPa, and extraction temperature 30-37 ℃, extraction time 150-240 minute, separator pressure 6-10MPa, separator temperature 40-55 ℃, carbon dioxide flow is 200-400L/H.
Beneficial effect of the present invention is: improve supercritical CO
2fluid extraction technology level, improves extraction yield, reduces production costs, so that supercritical CO
2fluid extraction technology is widely applied in propolis extracts.This method technique is simple, easy to operate, organic solvent-free, improve greatly production efficiency and can greatly improve the extraction content of flavone compound, this method make propolis super natural because of subconstiuent complete, particularly the content of the flavone aglycone such as myricetin, Quercetin, apiolin, pinocembrin is improved, and has strengthened the antioxidation activity of propolis extract, can better propolis be applied to the fields such as medicine, health food, cosmetics.
Accompanying drawing explanation
Fig. 1 is the TIC of hair glue after pretreatment (sample 1)
The TIC that Fig. 2 is the super natural factor of propolis (sample 2) that obtains through example one
The TIC that Fig. 3 is the super natural factor of propolis (sample 3) that obtains through example two
The specific embodiment
By specific embodiment, the specific embodiment of the present invention is described in further detail below.
Example one
A kind of solid enzymatic hydrolysis-CO
2the super natural factor of fluid supercritical extract propolis, comprises following processing step:
1, take and remove deimpurity hair glue 150kg, be placed in freezing 48h at-18 ℃ of temperature, adopt liquid nitrogen frozen to be ground into 10 object fine graineds (sample 1).
2, by hair collagen material weight ratio 1:1, add mixed material to be layered on after complex enzyme system and on biological enzymolysis bed, carry out solid enzymatic hydrolysis reaction, 50 ℃ of hydrolysis temperatures, enzymolysis humidity 35%, enzymolysis pH4.0, enzymolysis time 12h; Wherein, complex enzyme system is comprised of β-rhamnosidase (enzyme dosage 50IU/mg), beta-glucosidase (enzyme dosage 50IU/mg), zytase (enzyme dosage 1000IU/mg), pectase (enzyme dosage 20IU/mg) and AMS (enzyme dosage 10IU/mg).
3, the material after enzymolysis is complete is filled in extraction kettle, injects CO
2fluid extraction, extracting pressure 31MPa, 33 ℃ of extraction temperature, extraction time 240 minutes, separator pressure 8MPa, 48 ℃ of separator temperatures, carbon dioxide flow is 400L/H, after extraction, system pressure is reduced to normal pressure, and extract is separated out, and obtains the super natural factor of propolis (sample 2).
Example two
A kind of solid enzymatic hydrolysis-CO
2the super natural factor of fluid supercritical extract propolis, comprises following processing step:
1, take and remove deimpurity hair glue 150kg, be placed in freezing 48h at-18 ℃ of temperature, adopt liquid nitrogen frozen to be ground into 10 object fine graineds.
2, by hair collagen material weight ratio 1:1, add mixed material to be layered on after complex enzyme system and on biological enzymolysis bed, carry out Enzymic reaction, 60 ℃ of hydrolysis temperatures, enzymolysis humidity 50%, enzymolysis pH5.5, enzymolysis time 24h; Wherein, complex enzyme system is comprised of β-rhamnosidase (enzyme dosage 100IU/mg), beta-glucosidase (enzyme dosage 100IU/mg), zytase (enzyme dosage 1000IU/mg), pectase (enzyme dosage 50IU/mg) and AMS (enzyme dosage 50IU/mg).
3, the material after enzymolysis is complete is filled in extraction kettle, injects CO
2fluid extraction, extracting pressure 33MPa, 30 ℃ of extraction temperature, extraction time 240 minutes, separator pressure 6MPa, 45 ℃ of separator temperatures, carbon dioxide flow is 300L/H, after extraction, system pressure is reduced to normal pressure, and extract is separated out, and obtains the super natural factor of propolis (sample 3)
Constituent analysis:
Utilize GC-MS method to adopting solid enzymatic hydrolysis-CO
2the super natural factor of fluid supercritical extraction technique gained propolis is carried out constituent analysis.(sample 1 raw material, sample 2, sample 3)
Component list:
From above data analysis, can find out, propolis raw material after solid enzymatic hydrolysis again through CO
2the composition of the super natural factor of propolis of supercritical extract gained is more complete.Identify altogether out 85 kinds of compounds, wherein 79 qualitative, 6 do not detect, and detect content and account for 96.46% of total composition.Alkene class has 22 kinds, accounts for altogether 42.32% of total composition, more AR-curcumene 10.25% wherein, curcumene 6.22%, β-selinene 5.48%, α-selinene 2.31%, farnesene 2.93%, γ-cadinene 2.75%, β-guaiene 1.68%; Alcohols has 19 kinds, accounts for altogether 24.41% of total composition, more phenmethylol 4.9% wherein, benzyl carbinol 3.39%, sagittol 1.37%, α-eucalyptol 1.36%, guaiol 1.37%; Acids has 8 kinds, accounts for altogether 13.68% of total composition, more benzoic acid 4.6% wherein, 2-methyl-2-butenoic acid 4.29%, acetic acid 2.66%; Aldehydes has 8 kinds, accounts for altogether 6.4% of total composition, more benzaldehyde 1.93% wherein, cinnamic acid 1.23%; Ester class has 7 kinds, accounts for altogether 2.37% of total composition; In addition, the relative amount such as ether, alkane, ketone, phenols accounts for 7.28% of total composition altogether.
Here description of the invention and application is illustrative, not wants that therefore, the present invention is not subject to the restriction of the present embodiment by scope restriction of the present invention in the above-described embodiments, and the technical scheme that any employing equivalence replacement obtains is all in the scope of protection of the invention.
Claims (3)
1. a solid enzymatic hydrolysis-CO
2the method of the super natural factor of fluid supercritical extract propolis, is characterized in that, its preparation method is as follows:
(1), the pretreatment of hair glue: hair glue, through preliminary election classification, is removed impurity;
Put into refrigerator, be placed in freezing 48h at-18 ℃ of temperature, the hair glue after freezing, put into rapidly disintegrating machine mode broken or employing liquid nitrogen frozen and carry out fragmentation, be broken into 10-20 object particle, the raw material after fragmentation, enter immediately 20 object reciprocating sieves, be greater than 20 object particles and return to refrigerator;
(2), solid enzymatic hydrolysis: hair Jiao Peiyongshui and the complex enzyme system after crushing and screening is mixed, after mixed material is layered on solid enzymatic hydrolysis bed, regulate corresponding humiture, pH value, carry out solid enzymatic hydrolysis reaction;
(3), CO
2fluid supercritical extract: the propolis grain packing through solid enzymatic hydrolysis, in extraction kettle, is injected to CO
2fluid extracts, and after extraction, system pressure is reduced to normal pressure, and extract is separated out, and obtains the super natural factor of propolis.
2. a kind of solid enzymatic hydrolysis-CO as claimed in claim 1
2the method of the super natural factor of fluid supercritical extract propolis, it is characterized by, described solid enzymatic hydrolysis reaction enzymes is selected β-rhamnosidase, its enzyme dosage is 50-100IU/mg, beta-glucosidase, its enzyme dosage is 50-100IU/mg, zytase, its enzyme dosage is 1000-2000IU/mg, pectase, its enzyme dosage is 20-50IU/mg, AMS, its enzyme dosage is 10-50IU/mg or complex enzyme system, enzyme-added total amount is 0.5-1.5% by weight percentage, hydrolysis temperature 50-60 ℃, enzymolysis humidity 30-50%, enzymolysis pH 3.5-5.0, enzymolysis time 12-24h.
3. a kind of solid enzymatic hydrolysis-CO as claimed in claim 1
2the method of the super natural factor of fluid supercritical extract propolis, is characterized by, described CO
2the process conditions of fluid supercritical extract are as follows: extracting pressure 25-35MPa, and extraction temperature 30-37 ℃, extraction time 150-240 minute, separator pressure 6-10MPa, separator temperature 40-55 ℃, carbon dioxide flow is 200-400L/H.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114028429A (en) * | 2021-11-22 | 2022-02-11 | 江苏鸿祺生物科技有限公司 | Processing technology of propolis active component supercritical extraction raw material |
| CN114307235A (en) * | 2021-12-10 | 2022-04-12 | 江苏鸿祺生物科技有限公司 | Propolis with high flavone content and supercritical extraction process thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5529779A (en) * | 1991-08-27 | 1996-06-25 | Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo | Purified propolis-extract, and its preparation and uses |
| CN101307344A (en) * | 2008-07-08 | 2008-11-19 | 浙江大学 | Method for hydrolyzing propolis flavonoid glycosideby by naringinase |
| CN101530178A (en) * | 2009-04-16 | 2009-09-16 | 江苏江大源生态生物科技有限公司 | Method for extracting propolis by simulative biological fluid bed supercritical CO2 fluid |
| CN102321699A (en) * | 2011-06-24 | 2012-01-18 | 江苏江大源生态生物科技有限公司 | Method for composite enzymolysis-assisted extraction of active ingredient from propolis |
| CN102370667A (en) * | 2011-10-26 | 2012-03-14 | 江苏江大源生态生物科技有限公司 | Method for separating beeswax in propolis through supercritical CO2 |
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- 2013-11-21 CN CN201310593710.5A patent/CN103611333A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5529779A (en) * | 1991-08-27 | 1996-06-25 | Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo | Purified propolis-extract, and its preparation and uses |
| CN101307344A (en) * | 2008-07-08 | 2008-11-19 | 浙江大学 | Method for hydrolyzing propolis flavonoid glycosideby by naringinase |
| CN101530178A (en) * | 2009-04-16 | 2009-09-16 | 江苏江大源生态生物科技有限公司 | Method for extracting propolis by simulative biological fluid bed supercritical CO2 fluid |
| CN102321699A (en) * | 2011-06-24 | 2012-01-18 | 江苏江大源生态生物科技有限公司 | Method for composite enzymolysis-assisted extraction of active ingredient from propolis |
| CN102370667A (en) * | 2011-10-26 | 2012-03-14 | 江苏江大源生态生物科技有限公司 | Method for separating beeswax in propolis through supercritical CO2 |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114028429A (en) * | 2021-11-22 | 2022-02-11 | 江苏鸿祺生物科技有限公司 | Processing technology of propolis active component supercritical extraction raw material |
| CN114028429B (en) * | 2021-11-22 | 2022-12-16 | 江苏鸿祺生物科技有限公司 | Processing technology of propolis active component supercritical extraction raw material |
| CN114307235A (en) * | 2021-12-10 | 2022-04-12 | 江苏鸿祺生物科技有限公司 | Propolis with high flavone content and supercritical extraction process thereof |
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Application publication date: 20140305 |