CN104744600B - Process for performing high pressure combined ultrasound-associated extraction on active substance from mushroom - Google Patents
Process for performing high pressure combined ultrasound-associated extraction on active substance from mushroom Download PDFInfo
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Abstract
The invention provides a process for performing high pressure combined ultrasound-associated extraction on active substance from mushroom. The process sequentially comprises the following step: performing material preprocessing, pulverization, low-temperature lipid extraction, filter slag pressurization, ultrasound extraction and ethanol extraction, and finally drying to obtain the active substance. The method is characterized in that lipid extraction is adopted, the active substance dissolved with lipid is extracted, slag subjected to the lipid extraction adopts a high pressure combined ultrasound extraction technology to ensure that the difference between the internal pressure and the external pressure of the raw materials is changed to result in the change of a cell structure; then, an ultrasound-assisted extraction technology is adopted, an ultrasound cavatition function is utilized, target components are easily dissociated from an extracted matter, so that the active substance can be quickly and effectively extracted from the organism, thus greatly increasing the yield; then, the active substances subjected to lipid extraction and ethanol extraction are mixed and dried to prepare the active substance, and the active substance is stored at low temperature and the extraction rate of the active substance is greatly increased.
Description
Technical field
The present invention relates to a kind of extracting method to active substance in mushroom, specially a kind of high pressure combining ultrasonic auxiliary is carried
The technique for taking active substance in mushroom, belongs to biomedicine technical field.
Background technology
At present, the research for antioxidant and slow down aging medicine being separated from natural product causes the wide of Chinese scholars
General concern, has been achieved for more achievement in research, comprehensive research report both domestic and external, natural anti-oxidation activity in this field
Composition is mainly derived from the aspects such as plant, funguses (mushrooms), marine organisms.Wherein study it is wide be plant origin antioxygen
Change active component, there are some researches prove that the crude extract of the first alcohol and water of mushroom has free radical scavenging and suppresses Rat Erythrocytes haemolysis
Effect, the polysaccharide which contains can remove free radical, suppress lipid peroxidation of the superoxide anion to erythrocyte, and right
The lipid peroxidation of linoleic acid, vegetable oil oxidation and in vitro liver organization that free radical causes also has certain inhibitory action,
Therefore, mushrooms fruiting body extract and its polysaccharide resource are developed, therefrom screens the thing with antioxidant activity of nontoxic or low toxicity
Matter, has important practical significance and using value, it has also become the focus of research.
But the method extraction time that current mushroom active substance is extracted is longer, general a cycle needs for a long time, and carries
Take rate not high, cause wastage of material, it is impossible to rapidly and efficiently active substance is extracted from mushroom products so that further changing
Enter.
The content of the invention
For above-mentioned situation, it is the defect for solving prior art, the purpose of the present invention is just to provide a kind of high pressure joint and surpasses
The technique of active substance in sound assisted extraction mushroom, it is characterised in which comprises the following steps：
(1) pre-treatment of raw material：Raw material is carried out successively cleaning, cut into slices and negative pressure dehydrate；
(2) crush：The sliced mushroom after negative pressure dehydrate is rubbed with tissue mashing machine, and it is dry with negative pressure dehydration again
It is dry, obtain entity powder；
(3) low temperature fat is carried：Add petroleum ether according to a certain percentage, using blender stir certain hour, stand 8 hours with
On, supernatant is collected by filtration using filter paper, and repetitive operation 4-5 time, merge supernatant；
(4) filtering residue pressurization：Filtering residue under filter paper filter in step (3) is added into certain density sodium chloride solution, mixing is equal
Add after even in high pressure bag, sealing, and to Quick-pressing in the high pressure bag to 200-300MPa, then pressurize certain hour,
Then fast pressure relief, release repeats pressurization, pressurize and release after finishing, and operates 3-5 time；
(5) ultrasonic extraction：Solution in step (4) after treatment is poured in beaker, ultrasonic extraction instrument is inserted,
Ultrasonic extraction is carried out to solution, is then filtered using filter paper, is taken supernatant；
(6) ethanol is extracted：By obtained supernatant mixing, side stirring in the supernatant prepared in step (3) and step (5)
While the ethanol that mass concentration is 95% is added in mixed liquor, 2h is stood, then centrifugation a period of time on centrifuge, after filtration
It is precipitated；
(7) it is dried to obtain active substance：Negative pressure lyophilizing will be precipitated, the active substance of extraction needed for obtaining.
Further, preferably, concrete step is in step (1)：Raw material is wiped using gas or with totally
The mode that dry wipe is wiped is cleaned, and the thickness cut into slices during section ensures less than 3mm, and the temperature of negative pressure dehydrate keeps low
In 18 degrees Celsius, and the water content of section after being dried, is made to be less than 5.5%.
Further, preferably, step (2) crush in, mushroom tissue mashing machine rubbed using the sieves of 15 mesh
Mushroom powder is filtrated to get, and uses negative pressure dehydrate again, obtain entity powder of the water content less than 4%.
Further, preferably, the low temperature fat of step (3) is carried in step, the consumption for adding petroleum ether is entity powder
3-4 times, half an hour is first impregnated under 16 degrees Celsius, then stirred half an hour using blender, filter paper used is qualitative
Filter paper, filters during latter incorporated supernatant is placed on less than 18 degrees Celsius of environment and places stand-by.
Further, preferably, in the pressurization of step (4) filtering residue, the concentration of the sodium chloride solution of addition is 0.4mol/
LNaCl, and 1.4 times of the speed at least speed of the Quick-pressing to fast pressure relief in the high pressure bag.
Further, preferably, the speed of the pressurization is at least 65MPa/s, the decompression rate must not be higher than
300MPa/s。
Further, preferably, in the ultrasonic extraction step of the step (5), the power of ultrasound wave is 120-140W,
The ultrasonic wave concussion time is 20min, and, at least shakes 3 times every 5min ultrasonic vibrations once；Adopt filter paper filter number of times for
3-4 time.
Further, preferably, in step (6) ethanol extraction step：Mixed liquor and 95% ethanol volume ratio be 1
: between 3 to 1: 4, the rotating speed of centrifuge is 5000r/min, and centrifugation time is 5min, and is repeated at least the step of filtration after being centrifuged
3 times；
Further, preferably, being dried to obtain in active substance step in step (7), the temperature of the negative pressure lyophilizing is
Less than 5 degrees Celsius, and the active substance extracted needed for obtaining is put at a temperature of less than -10 degrees Celsius and preserves storage.
Beneficial effects of the present invention：The present invention is extracted initially with fat, is extracted the active substance for being dissolved in fat first,
Then the slag after fat is carried adopts high pressure combining ultrasonic extractive technique, raw material inside and outside differential pressure can be made to change, cause cell to be tied
Structure changes, and in conjunction with ultrasound assisted extraction, using ultrasonic cavitation, target components can be made to be more easy to from extract
Separate out, fast and efficiently can extract active substance from biological in vivo, and yield is greatly improved, then carried and ethanol fat
The active substance mixing of extraction, is dried, and active substance, and low temperature storage is obtained, and the extraction ratio for extracting active substance is greatly improved.
Specific embodiment
With reference to experiment, the present invention will be further described.
The invention provides in a kind of high pressure combining ultrasonic assisted extraction mushroom active substance technique, which includes following step
Suddenly：
(1) pre-treatment of raw material：Raw material is carried out clearly using gas erasing or by the way of being wiped with clean dry wipe
Wash, then cut into slices, the thickness cut into slices during section ensures less than 3mm, the temperature of negative pressure dehydrate keeps below 18 degrees Celsius, most
The water content of section is made to be less than 5.5% afterwards after negative pressure dehydrate, and drying
(2) crush：The sliced mushroom after negative pressure dehydrate is rubbed with tissue mashing machine, afterwards using the sieves of 15 mesh
Mushroom powder is filtrated to get to the mushroom that tissue mashing machine rubs, and uses negative pressure dehydrate again, water content is obtained less than 4%
Entity powder.
(3) low temperature fat is carried：Petroleum ether is added according to a certain percentage, and the consumption for adding petroleum ether is 3-4 times of entity powder,
Half an hour is first impregnated under 16 degrees Celsius, half an hour is stirred using blender then, stand more than 8 hours, using filter paper
It is collected by filtration supernatant, and repetitive operation 4-5 time, merge supernatant；Filter paper used is qualitative filter paper, is filtered on latter incorporated
Clear liquid places stand-by in being placed on less than 18 degrees Celsius of environment.
(4) filtering residue pressurization：Filtering residue under filter paper filter in step (3) is added into certain density sodium chloride solution, addition
The concentration of sodium chloride solution is 0.4mol/L NaCl, is added in high pressure bag, sealing after mix homogeneously, and into the high pressure bag
To 200-300MPa, then pressurize certain hour, then fast pressure relief, release repeat pressurization, pressurize to Quick-pressing after finishing
And release, and operate 3-5 time, and the speed for ensureing to pressurize is 70MPa/s, the decompression rate is 140MPa/s；
(5) ultrasonic extraction：Solution in step (4) after treatment is poured in beaker, ultrasonic extraction instrument is inserted,
Ultrasonic extraction is carried out to solution, the power of ultrasound wave is 120-140W, and the ultrasonic wave concussion time is 20min, and every 5min
Ultrasonic vibration once, at least shakes 3 times；Then filtered using filter paper, adopt the number of times that filter paper is filtered for 3 times, take supernatant；
(6) ethanol is extracted：By obtained supernatant mixing, side stirring in the supernatant prepared in step (3) and step (5)
While in mixed liquor add mass concentration be 95% ethanol, mixed liquor with 95% ethanol volume ratio for 1: 3 to 1: 4 it
Between, then centrifugation a period of time on centrifuge, the rotating speed of centrifuge is 5000r/min, and centrifugation time is 5min, and after being centrifuged
The step of filtration, repeats at least 3 times standing 2h, is precipitated after filtration；
(7) it is dried to obtain active substance：Negative pressure lyophilizing will be precipitated, the active substance of extraction needed for obtaining.And negative pressure lyophilizing
Temperature be that, less than 5 degrees Celsius, and the active substance extracted needed for obtaining is put in less than storage is preserved at a temperature of -10 degrees Celsius
Hide.
The present invention is extracted initially with fat, is extracted the active substance for being dissolved in fat first, and the slag after then fat is carried is adopted
With high pressure combining ultrasonic extractive technique, as high pressure is acted on, raw material inside and outside differential pressure can be made to change, cause cellularity to be sent out
Changing, in conjunction with ultrasound assisted extraction, using ultrasonic cavitation, can make target components be more easy to dissociate from extract
Out, active substance can be extracted in vivo from biological fast and efficiently, yield is greatly improved, and is then carried fat and being extracted with ethanol
Active substance mixing, be dried, be obtained active substance, and low temperature storage, extract active substance extraction ratio greatly improve.
Claims (4)
1. in a kind of high pressure combining ultrasonic assisted extraction mushroom active substance technique, it is characterised in which comprises the following steps：
(1) pre-treatment of raw material：Raw material is carried out successively cleaning, cut into slices and negative pressure dehydrate；Specially：Raw material is adopted
The mode wiped with gas or wiped with clean dry wipe is cleaned, and the thickness cut into slices during section ensures less than 3mm, negative pressure
The temperature of dehydrate keeps below 18 degrees Celsius, and makes the water content of section be less than 5.5% after being dried；
(2) crush：The sliced mushroom after negative pressure dehydrate is rubbed with tissue mashing machine, and use negative pressure dehydrate again, obtained
To entity powder；During step (2) is crushed, mushroom powder is filtrated to get to the mushroom that tissue mashing machine rubs using the sieves of 15 mesh
End, and negative pressure dehydrate is used again, obtain entity powder of the water content less than 4%；
(3) low temperature fat is carried：Petroleum ether is added according to a certain percentage, certain hour is stirred using blender, stand more than 8 hours,
Supernatant is collected by filtration using filter paper, and repetitive operation 4-5 time, merge supernatant；The low temperature fat of step (3) is carried in step, plus
The consumption for entering petroleum ether is 3-4 times of entity powder, first impregnates half an hour, then stirred using blender under 16 degrees Celsius
Half an hour, filter paper used are qualitative filter paper, filter during latter incorporated supernatant is placed on less than 18 degrees Celsius of environment and put
Put stand-by；
(4) filtering residue pressurization：Filtering residue under filter paper filter in step (3) is added into certain density sodium chloride solution, after mix homogeneously
Add in high pressure bag, sealing, and to Quick-pressing in the high pressure bag to 200-300MPa, then pressurize certain hour, then
Fast pressure relief, release repeat pressurization, pressurize and release after finishing, and operate 3-5 time；In the pressurization of step (4) filtering residue, addition
The concentration of sodium chloride solution is 0.4mol/L, and is at least the Quick-pressing to the speed of fast pressure relief in the high pressure bag
1.4 times of speed；The speed of the pressurization is at least 65MPa/s, and the decompression rate must not be higher than 300MPa/s；
(5) ultrasonic extraction：Solution in step (4) after treatment is poured in beaker, ultrasonic extraction instrument is inserted, to molten
Liquid carries out ultrasonic extraction, is then filtered using filter paper, takes supernatant；
(6) ethanol is extracted：By obtained supernatant mixing in the supernatant prepared in step (3) and step (5), while stirring to
The ethanol that mass concentration is 95% is added in mixed liquor, 2h is stood, then centrifugation a period of time on centrifuge, is obtained after filtration
Precipitation；
(7) it is dried to obtain active substance：Negative pressure lyophilizing will be precipitated, the active substance of extraction needed for obtaining.
2. in a kind of high pressure combining ultrasonic assisted extraction mushroom according to claim 1 active substance technique, its feature
It is that, in the ultrasonic extraction step of the step (5), the power of ultrasound wave is 120-140W, the ultrasonic wave concussion time is
20min, and, at least shake 3 times every 5min ultrasonic vibrations once；The number of times that filter paper is filtered is adopted for 3-4 time.
3. in a kind of high pressure combining ultrasonic assisted extraction mushroom according to claim 1 active substance technique, its feature
It is, in step (6) ethanol extraction step：Mixed liquor and 95% ethanol volume ratio be 1: 3 to 1: 4 between, centrifuge
Rotating speed be 5000r/min, centrifugation time is 5min, and be centrifuged after filter the step of repeat at least 3 times.
4. in a kind of high pressure combining ultrasonic assisted extraction mushroom according to claim 1 active substance technique, its feature
It is to be dried to obtain in active substance step in step (7), the temperature of the negative pressure lyophilizing is, less than 5 degrees Celsius, and will to obtain
The active substance of required extraction is put at a temperature of less than -10 degrees Celsius and preserves storage.
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| CN106892999A (en) * | 2017-04-24 | 2017-06-27 | 浙江海洋大学 | A kind of brown alga okamura ishige algae extraction method of polysaccharides |
| CN110881524A (en) * | 2018-09-11 | 2020-03-17 | 许昌神飞航天生物科技有限公司 | Solid beverage suitable for astronauts to maintain eyesight |
| CN110922494B (en) * | 2019-11-01 | 2021-09-21 | 福州福德恩生物科技有限公司 | Hericium erinaceus polysaccharide extraction and separation method |
| CN110903408B (en) * | 2019-11-01 | 2021-09-17 | 福州福德恩生物科技有限公司 | Preparation method of hericium erinaceus polysaccharide protein extract |
| CN110882291A (en) * | 2019-12-11 | 2020-03-17 | 睿侨(上海)生物科技有限公司 | Ultrasonic extraction process of fresh ginseng roots |
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