CN107964000A - The double assisted extraction beeswax flavones of enzyme-ultrasound - Google Patents
The double assisted extraction beeswax flavones of enzyme-ultrasound Download PDFInfo
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- CN107964000A CN107964000A CN201711214612.0A CN201711214612A CN107964000A CN 107964000 A CN107964000 A CN 107964000A CN 201711214612 A CN201711214612 A CN 201711214612A CN 107964000 A CN107964000 A CN 107964000A
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- Prior art keywords
- beeswax
- flavones
- extraction
- enzyme
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
- C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
- C07D311/28—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only
- C07D311/30—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only not hydrogenated in the hetero ring, e.g. flavones
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
- C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
- C07D311/40—Separation, e.g. from natural material; Purification
Abstract
The present invention discloses the double assisted extraction beeswax flavones of enzyme ultrasound, and step is as follows：Impurity elimination, drying are placed in pulverizer, are ground into powder；Mixing enzymatic treatment is first added, adds ethanol, ultrasonic extraction, is stirred, and is filtered, and takes supernatant fluid filtrate, and repeats to take filtrate to operate 2 times；It is concentrated under reduced pressure, n-hexane aqueous solution is extracted, grease removal, adds ethanol constant volume；Sodium nitrite solution is first added, adds 10% aluminum nitrate solution, is eventually adding 4% sodium hydroxide solution, scale is diluted to ethanol, 12min is stood after shaking up, absorbance is measured at 510nm.Extracting method of the present invention is simple, easily operated, extraction cost is low, under the extraction conditions of preferred process, the recovery rate of flavones is high, and active ingredient, organic solvent-free residual are not destroyed, purity is high, compared to extract by solvents, greatly shorten extraction time, the utilization rate of beeswax is effectively improved, so that beeswax maximum benefit, is adapted to industrialized production.
Description
Technical field
The present invention relates to the extracting method of flavones, and in particular to the double assisted extraction beeswax flavones of enzyme-ultrasound.
Background technology
Beeswax, also known as yellow wax, beeswax.Beeswax is the one kind being secreted by 4 pairs of wax glands of of the right age worker bee belly in bee colony
Fatty matter.In bee colony, worker bee is covered using the wax that oneself is secreted to build honeycomb, ovary capping and feed room.Honeycomb
It is to hoard food for honeybee, cultivate bee and inhabit the place of conglomeration, therefore, beeswax is both product and its existence of bee colony
With material necessary to breeding.Its main component of beeswax has:Acids, free fatty, free alkyl alcohol and carbohydrate.This
Outside, there are carotenoid, vitamin A, aromatic substance etc..But because of the differences such as peak kind, the sources of nectar and pollen, refinement method, its component
Also there is certain difference.
And flavones is also referred to as flavone, nature is widely present in, particularly in plant, due to the flavonoids found at first
Compound all has a ketone carbonyl group structure, and is in yellow or faint yellow, therefore claims flavones.Flavones has multiple biological activities, it
Physiological action paid close attention to always by people.Early 20th century, mid-term found that there is flavones Vitamin C activity and Anti-oxidant to live
Property, after the eighties, it is studied and starts to turn to the removing to active oxygen radical and the prevention to disease of old people functionally.It is many
Research has confirmed, flavones have anti-oxidant, antibacterial and it is antifatigue, remove free radical and the effect such as ridicule.Therefore, if can be by beeswax
Extracting flavonoids application, can greatly degree improve beeswax comprehensive utilization value.
The content of the invention
In view of the deficiencies of the prior art, the object of the present invention is to provide a kind of double assisted extraction beeswax flavones of enzyme-ultrasound, sheet
Extraction process cost is low, method is simple, easy to operate, recovery rate is high, and does not destroy active ingredient, organic solvent-free residual, pure
Degree is high, can make beeswax resource greatest benefit.
In order to achieve the above object, the present invention adopts the technical scheme that：
The double assisted extraction beeswax flavones of enzyme-ultrasound, comprise the following steps：
(1) beeswax impurity is removed, drying is placed in pulverizer, is crushed into powder last spare；
(2) powder into step (1), first adds the mixing enzymatic treatment 1h of cellulase and pectase composition, and described mixed
The mass ratio of synthase cellulase and pectase is 1：1~1:2, then by solid-liquid ratio be 1：20g/ml adds volume fraction
60% ethanol extractant, be placed in power be 60W ultrasonic cleaner in, at a temperature of 45~55 DEG C extract 20~
30min, and repeat extraction 3 times；
(3) after treating step (2) extraction, extracting solution is stirred and is filtered, take supernatant fluid filtrate, and repeat to take filtrate
Operation 2 times, takes supernatant fluid filtrate to be concentrated under reduced pressure, and n-hexane aqueous solution is extracted, grease removal, and 60% ethanol constant volume of heating is until dilution
5 times；
(4) sample diluting liquid in step (3) is taken, first adds 5% sodium nitrite solution, 6min is stood after shaking up, adds
10% aluminum nitrate solution, also stands 6min after shaking up, be eventually adding 4% sodium hydroxide solution, and scale is diluted to 60% ethanol,
12min is stood after shaking up, absorbance is measured at 510nm.
As optimal technical scheme, in order to provide rational mixing enzyme component, the flavones in beeswax is set to be fully dissolved in second
In alcohol extracting agent, to be conducive to improve the recovery rate and purity of beeswax flavones, while the waste of resource is avoided, the step (2)
The mass ratio of mixed enzyme cellulase and pectase is 1：1.
As optimal technical scheme, in order to provide suitable Extracting temperature, the flavones in beeswax is set all to dissolve and extract, with
Be conducive to improve the recovery rate and purity of beeswax flavones, while effectively control extraction cost, ultrasonic extraction temperature in the step (2)
Spend for 50 DEG C.
As optimal technical scheme, in order to provide the reasonable drawing time, flavones is set to be dissolved in as far as possible in ethanol extractant,
To be conducive to improve the recovery rate and purity of beeswax flavones, while the waste of resource is avoided, step (2) the ultrasonic extraction time
For 25min.
Beneficial effects of the present invention：Extracting method of the present invention is simple, easily operated, extraction cost is low, in cellulase and
The mass ratio of pectase is 1：1st, ultrasonic temperature be 50 DEG C, ultrasonic power 60W, ultrasonic time 25min, extraction time 3
Under the extraction conditions of secondary preferred process, the recovery rate of flavones is high, and does not destroy active ingredient, organic solvent-free residual, and purity is high,
Compared to extract by solvents, extraction time is greatly shortened, effectively improves the utilization rate of beeswax, so that beeswax maximum benefit,
It is adapted to industrialized production.
Embodiment
The present invention is further described with reference to specific embodiment.
Embodiment 1
The double assisted extraction beeswax flavones of enzyme-ultrasound, comprise the following steps：
(1) beeswax impurity is removed, drying is placed in pulverizer, is crushed into powder last spare；
(2) powder into step (1), first adds the mixing enzymatic treatment 1h of cellulase and pectase composition, and described mixed
The mass ratio of synthase cellulase and pectase is 1：1, then by solid-liquid ratio be 1：It is 60% that 20g/ml, which adds volume fraction,
Ethanol extractant, is placed in the ultrasonic cleaner that power is 60W, 2min is extracted at a temperature of 45, and repeat extraction 3 times；
(3) after treating step (2) extraction, extracting solution is stirred and is filtered, take supernatant fluid filtrate, and repeat to take filtrate
Operation 2 times, takes supernatant fluid filtrate to be concentrated under reduced pressure, and n-hexane aqueous solution is extracted, grease removal, and 60% ethanol constant volume of heating is until dilution
5 times；
(4) sample diluting liquid in step (3) is taken, first adds 5% sodium nitrite solution, 6min is stood after shaking up, adds
10% aluminum nitrate solution, also stands 6min after shaking up, be eventually adding 4% sodium hydroxide solution, and scale is diluted to 60% ethanol,
12min is stood after shaking up, absorbance is measured at 510nm.
Embodiment 2
The double assisted extraction beeswax flavones of enzyme-ultrasound, comprise the following steps：
(1) beeswax impurity is taken out, drying is placed in pulverizer, is crushed into powder last spare；
(2) powder into step (1), first adds the mixing enzymatic treatment 1h of cellulase and pectase composition, and described mixed
The mass ratio of synthase cellulase and pectase is 1:2, then by solid-liquid ratio be 1：It is 60% that 20g/ml, which adds volume fraction,
Ethanol extractant, is placed in the ultrasonic cleaner that power is 60W, 25min is extracted at a temperature of 50 DEG C, and repeat extraction 3
It is secondary；
(3) after treating step (2) extraction, extracting solution is stirred and is filtered, take supernatant fluid filtrate, and repeat to take filtrate
Operation 2 times, takes supernatant fluid filtrate to be concentrated under reduced pressure, and n-hexane aqueous solution is extracted, grease removal, and 60% ethanol constant volume of heating is until dilution
5 times；
(4) sample diluting liquid in step (3) is taken, first adds 5% sodium nitrite solution, 6min is stood after shaking up, adds
10% aluminum nitrate solution, also stands 6min after shaking up, be eventually adding 4% sodium hydroxide solution, and scale is diluted to 60% ethanol,
12min is stood after shaking up, absorbance is measured at 510nm.
Embodiment 3
The double assisted extraction beeswax flavones of enzyme-ultrasound, comprise the following steps：
(1) beeswax impurity is taken out, drying is placed in pulverizer, is crushed into powder last spare；
(2) powder into step (1), first adds the mixing enzymatic treatment 1h of cellulase and pectase composition, and described mixed
The mass ratio of synthase cellulase and pectase is 1：1, then by solid-liquid ratio be 1：It is 60% that 20g/ml, which adds volume fraction,
Ethanol extractant, is placed in the ultrasonic cleaner that power is 60W, 20~30min is extracted at a temperature of 45~55 DEG C, and again
Extract 3 times again；
(3) after treating step (2) extraction, extracting solution is stirred and is filtered, take supernatant fluid filtrate, and repeat to take filtrate
Operation 2 times, takes supernatant fluid filtrate to be concentrated under reduced pressure, and n-hexane aqueous solution is extracted, grease removal, and 60% ethanol constant volume of heating is until dilution
5 times；
(4) sample diluting liquid in step (3) is taken, first adds 5% sodium nitrite solution, 6min is stood after shaking up, adds
10% aluminum nitrate solution, also stands 6min after shaking up, be eventually adding 4% sodium hydroxide solution, and scale is diluted to 60% ethanol,
12min is stood after shaking up, absorbance is measured at 510nm.
According to but Factor Experiment structure, design the species and ratio of enzyme, orthonormal design of experiments.Experimental data and result are as follows
Shown in table 1：
The influence table of 1 enzyme class of table and ratio to extracting flavonoids yield
Found according to experimental result, ultrasound assisted extraction beeswax flavones substantially reduces extraction time, and the addition of enzyme carries
The high extraction yield of beeswax flavones, the optimal extract process of beeswax flavones are：Extracting flavones optimum process condition is:
Ultrasonic temperature is 50 DEG C, ultrasonic power 60W, ultrasonic time 25min.
It should be pointed out that above case is only the representational example of the present invention.Obviously, technical scheme and unlimited
In above-described embodiment, there can also be many denaturation.
Claims (4)
1. the double assisted extraction beeswax flavones of enzyme-ultrasound, it is characterised in that comprise the following steps：
(1) beeswax impurity is taken out, drying is placed in pulverizer, is crushed into powder last spare；
(2) powder into step (1), first adds the mixing enzymatic treatment 1h of cellulase and pectase composition, and the mixed enzyme
The mass ratio of cellulase and pectase is 1：1~1:2, then by solid-liquid ratio be 1：It is 60% that 20g/ml, which adds volume fraction,
Ethanol extractant, is placed in the ultrasonic cleaner that power is 60W, 20~30min is extracted at a temperature of 45~55 DEG C, and again
Extract 3 times again；
(3) after treating step (2) extraction, extracting solution is stirred and is filtered, take supernatant fluid filtrate, and repeat to take filtrate to operate
2 times, supernatant fluid filtrate is taken to be concentrated under reduced pressure, n-hexane aqueous solution is extracted, grease removal, and 60% ethanol constant volume of heating is until 5 times of dilution；
(4) sample diluting liquid in step (3) is taken, first adds 5% sodium nitrite solution, 6min is stood after shaking up, adds 10%
Aluminum nitrate solution, also stands 6min after shaking up, be eventually adding 4% sodium hydroxide solution, is diluted to scale with 60% ethanol, shakes up
After stand 12min, at 510nm measure absorbance.
2. the double assisted extraction beeswax flavones of enzyme-ultrasound according to claim 1, it is characterised in that：Step (2) mixing
The mass ratio of enzyme cellulase and pectase is 1：1.
3. the double assisted extraction beeswax flavones of enzyme-ultrasound according to claim 2, it is characterised in that：Surpass in the step (2)
Sound Extracting temperature is 50 DEG C.
4. the double assisted extraction beeswax flavones of enzyme-ultrasound according to claim 3, it is characterised in that：Step (2) ultrasound
Extraction time is 25min.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201711214612.0A CN107964000A (en) | 2017-11-28 | 2017-11-28 | The double assisted extraction beeswax flavones of enzyme-ultrasound |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201711214612.0A CN107964000A (en) | 2017-11-28 | 2017-11-28 | The double assisted extraction beeswax flavones of enzyme-ultrasound |
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| CN107964000A true CN107964000A (en) | 2018-04-27 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111440218A (en) * | 2020-04-24 | 2020-07-24 | 上海珈凯生物科技有限公司 | Preparation method of plant polyphenol |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1465569A (en) * | 2002-06-25 | 2004-01-07 | 杨汝伟 | Method for raising total flavone content in propolis |
| CN101082055A (en) * | 2006-05-31 | 2007-12-05 | 新疆农业科学院微生物应用研究所 | Method for extracting flavonoids substance from liquorice slag by employing complex enzyme process |
| JP2009254286A (en) * | 2008-04-16 | 2009-11-05 | Kao Corp | Method for producing flavor oil |
-
2017
- 2017-11-28 CN CN201711214612.0A patent/CN107964000A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1465569A (en) * | 2002-06-25 | 2004-01-07 | 杨汝伟 | Method for raising total flavone content in propolis |
| CN101082055A (en) * | 2006-05-31 | 2007-12-05 | 新疆农业科学院微生物应用研究所 | Method for extracting flavonoids substance from liquorice slag by employing complex enzyme process |
| JP2009254286A (en) * | 2008-04-16 | 2009-11-05 | Kao Corp | Method for producing flavor oil |
Non-Patent Citations (1)
| Title |
|---|
| 蔡晓东等: "蜂蜡黄酮提取工艺研究", 《现代食品》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111440218A (en) * | 2020-04-24 | 2020-07-24 | 上海珈凯生物科技有限公司 | Preparation method of plant polyphenol |
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