CN107334096A - A kind of method and its extract that active component is extracted from sea cucumber - Google Patents

A kind of method and its extract that active component is extracted from sea cucumber Download PDF

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Publication number
CN107334096A
CN107334096A CN201710677091.6A CN201710677091A CN107334096A CN 107334096 A CN107334096 A CN 107334096A CN 201710677091 A CN201710677091 A CN 201710677091A CN 107334096 A CN107334096 A CN 107334096A
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sea cucumber
freeze
extract
active component
ultrasonication
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董汉玉
徐明恺
董佳琪
郭巍
刘昌孝
李梦虹
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Tai Wan Han Wei (Guangzhou) Pharmaceutical Technology Co., Ltd.
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Hebei Sino Biological Pharmaceutical Co Ltd
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
    • A23L17/20—Fish extracts

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  • Marine Sciences & Fisheries (AREA)
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  • Health & Medical Sciences (AREA)
  • Nutrition Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

The invention discloses a kind of method and its extract that active component is extracted from sea cucumber.Described extracting method comprises the following steps：Sea cucumber slurry is ultrasonically treated；The temperature of the processing is 0 DEG C 10 DEG C.This method is sufficiently extracted fat-soluble higher sea cucumber unrighted acid, vitamin E etc. and water-soluble higher holothurian collagen, sea cucumber polysaccharide, sea cucumber polypeptide isoreactivity material in sea cucumber respectively as far as possible, Holothurian Resources are maximally utilized, while eliminate the small molecule contaminants matter such as heavy metal ion of residual in sea cucumber body；Remain the activity of active ingredient in sea cucumber to greatest extent using gentle physics and biological method, prevent active component loss in process and activity change.

Description

A kind of method and its extract that active component is extracted from sea cucumber
Technical field
The present invention relates to a kind of method and its extract that active component is extracted from sea cucumber.
Background technology
Sea cucumber, belong to Echinodermata (Echinodenmata) Holothuroidea (Holothuroidea), be to live by the sea extremely 8000 meters of ocean echinoderm, it is widely distributed in each ocean in the world, away from modern existing more than 600,000,000 years history.The whole world there are about at present Sea cucumber kind 1100, China's Coastline of South China Sea species is more, and there are about more than 20 kind sea cucumbers can be edible.Sea cucumber is not only preciousness Food, and rare medicinal material, the same ginseng of sea cucumber, bird's nest, shark's fin are equally celebrated for their achievements, are one of eight big treasures of the world.According to《Compendium of Materia Medica picks up Lose》Described in：Sea cucumber, sweet-salty, kidney tonifying, benefiting quintessence, urine is taken the photograph, impotence, its warm-natured benefit are treated in establishing-Yang, and foot supports ginseng, therefore named sea cucumber. Sea cucumber have improve memory, delay sexual gland aging, prevent artery sclerosis and it is antitumor wait act on.
With the progress of modern medical science and the development of advanced detection means, the nutritive value and health-care efficacy of sea cucumber are got over More to be found by people, and approved by increasing people.Collagen in sea cucumber, is easily absorbed by human body, is Fabulous nutrient, and there are high beauty functions.Contain great amount of soluble sea cucumber polysaccharide, Sea Cucumber Glycosides etc. in sea cucumber Active material, have and improve immunity, antitumor, radiation-resistant effect well.And a large amount of fat-soluble things contained in sea cucumber Matter such as vitamin E, polyunsaturated fatty acid, antioxidase etc., can be effectively protected cardiovascular function, it is antifatigue, anti-oxidant, Anti-aging.
At present, it is still based on directly eating that the main sea cucumber of people, which takes mode,.Directly edible mode, causes sea Join the absorptivity deficiency in human body.And all traditional sea cucumber cooking methods, such as thermophilic digestion, also cause sea cucumber nutrient composition It is a large amount of to be lost in, cause to treasure the waste of resource.Such as the traditional approach of thermophilic digestion, not only make a large amount of water miscible active materials such as Sea cucumber polysaccharide and Sea Cucumber Glycosides etc. are dissolved among working fluid and discarded in vain, more cause the nutrients of a large amount of thermolabilities Matter, such as vitamin, antioxidase completely lose original physiologically active.Therefore, how the precious sea cucumber of processing of science, most The nutritive value of the reservation sea cucumber of big degree, it is always the target that nutritionist and foods mfr pursue.
The content of the invention
Problem to be solved by this invention is largely flowed for sea cucumber nutrient composition be present in existing sea cucumber extracting method Lose, extract it is insufficient, have metal residual, and provide it is a kind of from sea cucumber extract active component method and its extract.Should Method makes water-soluble and liposoluble active nutriment all obtain maximum reservation, improves the edibility and medicine of sea cucumber With value.
The present invention solves above-mentioned technical problem using following technical proposals：
The invention provides a kind of method that active component is extracted from sea cucumber, it comprises the following steps：By sea cucumber slurry It is ultrasonically treated, produces sea cucumber active component extract；The temperature of the ultrasonication is 0 DEG C -10 DEG C.
Described sea cucumber slurry can be obtained by sea cucumber after pretreatment.Described pretreatment may include following step：
(a) by sea cucumber impurity elimination, sea cucumber skin is obtained；
(b) described sea cucumber skin is crushed, the sea cucumber skin crushed；
(c) the sea cucumber skin of described crushing is ground together with water, obtains described sea cucumber slurry.
In described step (a), described sea cucumber is the conventional edible sea cucumber in this area, such as extra large stichopus japonicus (Stichopus japonicus)。
In described step (a), described impurity elimination is the conventional impurity elimination in this area, such as is removed the internal organs and silt.
In described step (a), the operation with water rinse is may also include after described impurity elimination.The behaviour of described water rinse Preferably include the following steps：Cleaned with clean running water, then cleaned with deionized water or distilled water.Time of described cleaning Number can be not especially limited, typically that the cleaning sea cucumber after impurity elimination is clean.
In described step (a), when may also include the operation with water rinse after described impurity elimination, the water moistens Also include draining operation after the operation washed.
In described step (b), described crushing is the conventional crushing in this area, is crushed for example with meat grinder.
In described step (c), the mass values of described water and the sea cucumber skin of described crushing are conventional for this area Mass values, such as 3~5, in another example 4.
In described step (c), described is ground to the conventional grinding in this area, is ground for example with high speed homogenizer Or colloid mill grinding.
In described step (c), the number of described grinding is the conventional number in this area, such as 1 time or 2 times.
In described step (c), the fineness of the sea cucumber slurry obtained after described grinding is the conventional fineness in this area, Such as 100~300 mesh, in another example 200 mesh.
Treatment temperature in described ultrasonication is preferably 4 DEG C.
In the present invention, the condition of described ultrasonication carries out conventional choosing according to the instrument of selected supersound process Select.For example, equivalent is few, during using ultrasonic cell disruptor, ultrasound condition is that transmitter power is 500W, the working time 10 Second, 15 seconds off times, work times 80 times.10kg is up to due to measuring larger, fresh sea cucumber in the present invention, thus it is of the invention In, described ultrasonication is preferably carried out in vacuum ultrasonic reactor.Described ultrasonication is so that sea cucumber slurry Vortex is produced, sea cucumber cell membrane obtains fully broken be defined.The time of described ultrasonication is preferably 150-200 minutes, More preferably 180 minutes.Power is preferably 300-700W in described ultrasonication, more preferably 500W.Described ultrasound Tranmitting frequency is preferably 20-30KHz in ripple processing, more preferably 25KHz.Pulse cycle frequency is excellent in described ultrasonication Elect 5-15 times/second, more preferably 10 times/second as.Acoustic beam is preferably 1200-2200 meter per seconds in described ultrasonication, more excellent Elect 1700 meter per seconds as.Liquid surface sound pressure level is preferably 15-55 ° in described ultrasonication, more preferably 20 °.Described Vacuum is preferably 80-120Pa in ultrasonication, more preferably 100Pa.
It is preferred that described sea cucumber active component extract is stored at -30 to -50 DEG C, more preferably in -40 DEG C of storages.
Present invention also offers a kind of sea cucumber active component extract, and it extracts active component according to above-mentioned from sea cucumber Method is made, and it is freeze-dried after obtain.The temperature of described freeze-drying is preferably -30 to -50 DEG C, and more preferably -40 ℃.The vacuum of described freeze-drying is preferably 10-20Pa, more preferably 15Pa；The time of described freeze-drying is preferred For 8-12 hours, more preferably 10 hours.Described freeze-drying is carried out preferably in freeze drier.Cold described in progress Before lyophilized dry operation, described freeze drier first preferably is carried out into pre-freeze, and (such as pre-freeze is to 0 DEG C -10 DEG C, preferably 4 ℃)。
Present invention also offers a kind of method that active component is extracted from sea cucumber, it comprises the following steps：
(1) sea cucumber slurry is ultrasonically treated；Obtain sea cucumber active component extract；Described treatment temperature is 0℃-10℃。
(2) by described sea cucumber active component extract by being centrifuged at a high speed to obtain solid-phase component and liquid phase ingredient.
In step (1), each method and condition are same as above.
In step (2), described being centrifuged at a high speed preferably on supercentrifuge is carried out.Described is ultracentrifugal Time is preferably 30-50 minutes, more preferably 45 minutes.Described ultracentrifugal revolution is preferably 10,000-15, 000rpm, more preferably 12,000rpm.
The described method that active component is extracted from sea cucumber may also include the behaviour handled described liquid phase ingredient Make.The processing of described liquid phase ingredient preferably includes the following steps：The behaviour that described liquid phase ingredient is passed through into filtering with microporous membrane Make, to remove solid impurity.Described miillpore filter aperture is preferably 0.22~0.8 μm, more preferably 0.45 μm.Described is micro- The pressure during membrane filtration of hole is preferably 0.5-1.0MPa, more preferably 0.8MPa.
The processing of described liquid phase ingredient may also include the following steps：Pass through NF membrane after described filtering with microporous membrane The operation of nanofiltration, with desalination and removing heavy metals ion.The pressure of described nanofiltration is preferably 1.0-1.8MPa, more preferably 1.5MPa.The aperture of described NF membrane is preferably 500-1,500 dalton, more preferably 1,000 dalton.Described liquid phase The processing of composition can also further comprise the following steps：It is freeze-dried after described nanofiltration.Described freeze-drying Temperature is preferably -30 to -50 DEG C, more preferably -40 DEG C.The vacuum of described freeze-drying is preferably 10-20Pa, more preferably For 15Pa；The time of described freeze-drying is preferably 8-12 hours, more preferably 10 hours.
The processing of described liquid phase ingredient can also further comprise being crushed after described freeze-drying, (example of sieving Such as 60 mesh), that is, obtain the sea cucumber extract of water-soluble portion.Described crushing preferably crushes in pulverizer.
It is preferred that it can also be comprised the steps of between described nanofiltration and freeze-drying：Concentrated after described nanofiltration Fall 60-80% moisture (such as 70% moisture), obtain concentrate, then be freeze-dried, described percentage refers to water Quality account for the percentage of described liquid phase ingredient gross mass.
In a preferred embodiment of the invention, the method that active component is extracted from sea cucumber, including following steps Suddenly (or being made up of following step)：
(a) sea cucumber slurry is ultrasonically treated；Obtain sea cucumber active component extract；Described treatment temperature is 0℃-10℃；
(b) by described sea cucumber active component extract by being centrifuged at a high speed to obtain solid-phase component and liquid phase ingredient；
(c) described liquid phase ingredient NF membrane nanofiltration, is freeze-dried, crushed by filtering with microporous membrane, you can.
In step (a)-(c), each condition and parameter are the same as those described above.
The described method that active component is extracted from sea cucumber may also include the processing to described solid-phase component.Described The processing of solid-phase component preferably includes the following steps：By described solid-phase component, CO is carried out2Supercritical extract, produce non-aqueous The sea cucumber composition of property part.It is preferred that by described solid-phase component after freeze-drying and crushing, thick sea cucumber powder is obtained, then CO is carried out again2Supercritical extract.Described freeze-drying and the condition of crushing are the same as the lyophilized dry and crushing of above-mentioned liquid phase ingredient Condition.Described CO2The preferred dynamic extraction of supercritical extract.Described CO2The pressure of supercritical extract is preferably 20-30MPa, More preferably 25MPa.Described CO2The temperature of supercritical extract is preferably 30-50 DEG C, more preferably 40 DEG C.Described CO2It is super CO in critical extraction2Flow velocity is preferably 20-30L/h, more preferably 25L/h.Described CO2The time of supercritical extract is preferably 2-3 hours.
In a preferred embodiment of the invention, the method that active component is extracted from sea cucumber, it includes following Step (or being made up of following step)：
(i) sea cucumber slurry is ultrasonically treated；Obtain sea cucumber active component extract；Described treatment temperature is 0℃-10℃；
(ii) will sea cucumber active component extract by being centrifuged at a high speed to obtain solid-phase component and liquid phase ingredient；
(iii) described solid-phase component is freeze-dried；Crush, obtain thick sea cucumber powder；
(iv) described thick sea cucumber powder is subjected to CO2Supercritical extract, you can.
In step (i)~(iv), each condition and parameter are the same as those described above.
Present invention also offers one kind sea cucumber active component extract as made from the above method.
Present invention also offers a kind of sea cucumber extract, and it is made by following methods, and described method comprises the following steps：
(1) sea cucumber slurry is ultrasonically treated；Obtain sea cucumber active component extract；Described treatment temperature is 0℃-10℃。
(2) by described sea cucumber active component extract by being centrifuged at a high speed to obtain liquid phase ingredient.
In a preferred embodiment of the invention, described from sea cucumber extract, it is made by following methods, described side Method comprises the steps (or being made up of following step)：
(a) sea cucumber slurry is ultrasonically treated；Obtain sea cucumber active component extract；Described treatment temperature is 0℃-10℃；
(b) by described sea cucumber active component extract by being centrifuged at a high speed to obtain solid-phase component and liquid phase ingredient；
(c) described liquid phase ingredient NF membrane nanofiltration, is freeze-dried, crushed by filtering with microporous membrane, you can.
Present invention also offers a kind of sea cucumber extract, and it is made by following methods, and described method comprises the following steps：
(1) sea cucumber slurry is ultrasonically treated；Obtain sea cucumber active component extract；Described treatment temperature is 0℃-10℃；
(2) by described sea cucumber active component extract by being centrifuged at a high speed to obtain solid-phase component.
In a preferred embodiment of the invention, described sea cucumber extract, it is made by following method, described method Comprise the steps (or being made up of following step)：
(i) sea cucumber slurry is ultrasonically treated；Obtain sea cucumber active component extract；Described treatment temperature is 0℃-10℃；
(ii) will sea cucumber active component extract by being centrifuged at a high speed to obtain solid-phase component and liquid phase ingredient；
(iii) described solid-phase component is freeze-dried；Crush, obtain thick sea cucumber powder；
(iv) described thick sea cucumber powder is subjected to CO2Supercritical extract, you can.
In the present invention, the different methods that active component is extracted from sea cucumber, obtain different sea cucumber activity into Divide extract.
Without prejudice to the field on the basis of common sense, above-mentioned each optimum condition, can be combined, and it is each preferably to produce the present invention Example.
Agents useful for same and raw material of the present invention are commercially available.
The positive effect of the present invention is：Fat-soluble higher sea cucumber in sea cucumber is sufficiently extracted respectively as far as possible Unrighted acid, vitamin E etc. and water-soluble higher holothurian collagen, sea cucumber polysaccharide, sea cucumber polypeptide isoreactivity material, Holothurian Resources are maximally utilized, while eliminate the small molecule contaminants matter such as heavy metal ion of residual in sea cucumber body； Remain the activity of active ingredient in sea cucumber to greatest extent using gentle physics and biological method, prevent active component from adding Loss and activity change during work；The processing method of the present invention has fully demonstrated advanced, feasible, efficient, energy-saving and environmental protection Feature, to realize that the higher value application of sea cucumber provides technical support.
Embodiment
The present invention is further illustrated below by the mode of embodiment, but does not therefore limit the present invention to described reality Apply among a scope.The experimental method of unreceipted actual conditions in the following example, conventionally and condition, or according to business Product specification selects.
In following embodiments and comparative example, such as it is not specifically noted, fresh extra large stichopus japonicus weight is 10kg.Each operation step Suddenly, such as it is not specifically noted, each means and operate at ambient temperature.
In following embodiments, described liquid phase ingredient is also known as water-soluble portion；Described solid-phase component is also known as water-insoluble Part.
The method that embodiment 1 extracts active component from sea cucumber
1) fresh extra large stichopus japonicus (Stichopus japonicus) is taken, removes clean internal organ, removes residual in intestinal wall Silt, it is put into rinse bath, with clean running water cleaning three times, adds deionized water cleaning once, pull out and drain away the water, Obtain sea cucumber skin.Start meat grinder, sea cucumber skin is rubbed one by one, the sea cucumber skin after rubbing is collected in stainless steel cask, adds 4 times of weights The deionized water of amount, is thoroughly mixed, and forms broken sea cucumber skin mixed liquor.Start colloid mill, setting fineness is 200 mesh, will The feeding mouth that broken sea cucumber mixed liquor gradually puts into colloid mill is ground, and the feed liquid of outlet grinds one repeatedly with the same manner again It is secondary, always it is co-mulled and made into twice, the sea cucumber slurry after finally being ground.
2) optimization of ultrasonication condition and ultrasonic wave extraction
A. the sea cucumber slurry obtained in 20ml steps 1) is taken, is carried out using JY92-II types ultrasonic cell disruptor small Scale ultrasonication is tested.The condition and parameter of ultrasonication are set as follows according to instrument specification：Transmitter work( Rate is 500W, 10 seconds working times, 15 seconds off times, and (due to being lab scale, ultrasound condition enters according to instrument for work times 80 times Row selection).The temperature of ultrasonication is controlled, respectively at 4 DEG C, 20 DEG C, 40 DEG C, 60 DEG C, 80 DEG C.After ultrasonication terminates, Carry out the assay of collagen.
B. the assay of collagen uses L- hydroxyproline indirect quantification methods：Concrete operations are：Weigh and be purchased from Sigma The L- hydroxyproline standard product of company, the L- hydroxyproline standard solution of various concentrations is configured with distilled water, with spectrophotometer Absorbance is surveyed at 558nm, draws L- hydroxyproline standard curves.
C. sea cucumber handles the measure of collagen content in sample：Weigh (the sea cucumber activity of the sample after treatment of different temperature Component extract) each 1ml, it is respectively put into ampoule bottle, adds 2mL 6mol/L HCl, alcohol blast burner sealing, 130 DEG C of hydrolysis 4h, it is transferred in beaker and adds 1 0.1% methyl red indicator of drop, 2mol/L NaOH adjusts pH to be settled to 100mL to neutrality, survey Absorbance, apply mechanically normal equation and calculate L- hydroxyproline contents.According to Saito document reports (Satto M, Kunisaki N, Urano N.Collagen as the major rdible component of sea cucumber(Stichopus japonicus).Journal of Food science,2002,67:1319-1322), L- hydroxyl dried meat in aquatic products collagen Histidine content is about 8.1% (mass fraction), therefore can draw collagen content by conversion.As a result as shown in table 1, can See, under the conditions for the treatment of temperature is elevated, the collagen content in sea cucumber active component extract significantly reduces, and handles temperature Degree is higher, and collagen content is lower.
Under the different temperatures of table 1. after ultrasonication sea cucumber slurry collagen content
Treatment temperature (DEG C) 4 20 40 60 80
Collagen content (mg/ml) 9.32±0.11 6.51±0.09 6.36±0.07 4.78±0.23 3.06±0.34
From the disclosure of table 1, when supersound process temperature is 4 DEG C, collagen in sea cucumber active component extract Content highest.
By in the above-mentioned sea cucumber slurry input vacuum ultrasonic reactor prepared, 100Pa is evacuated to, controls temperature in kettle 4 DEG C of degree, the power for adjusting ultrasonic transmitter is 500W, sets ultrasonic wave tranmitting frequency as 25KHz, pulse cycle frequency 10 Secondary/second, the meter per second of acoustic beam 1700, liquid surface sound pressure level are 20 °, start ultrasonic wave program, control ultrasonic intensity, make sea cucumber material Slurry produces obvious vortex.Ultrasonication 180 minutes, sea cucumber histocyte wall is set fully to be crushed, sea cucumber active component obtains To discharge, the sea cucumber slurry after ultrasonication is obtained.
Sea cucumber slurry after ultrasonication carries out freeze-drying and removes moisture removal.First freeze dryer is carried out before freeze-drying pre- It is cold, when freeze-drying machine partition board temperature is down to 4 DEG C, the stainless steel tray for the sea cucumber concentrate crossed in -40 DEG C of pre-freezes will be filled It is placed on shelf, it is -40 DEG C to set freeze-drying machine partition board temperature, opens water vessel refrigerating state, opens vavuum pump, sets vacuum For 15Pa, persistently freeze 10 hours.
Product after lyophilized crushes 30 minutes in pulverizer, crosses 100 mesh sieves, obtains sea cucumber active component extract.
3) by the sea cucumber slurry after ultrasonication, adding in the centrifuge tube of supercentrifuge in batches, set high The revolution of fast centrifuge is 12,000rpm, and centrifugation time 45 minutes, temperature is set as 4 DEG C, starts centrifuge and starts to centrifuge Journey.After centrifugation terminates, careful collection centrifuged supernatant, i.e. liquid phase ingredient；It is placed in a clean stainless steel cask.It is remaining solid Phase constituent is collected in stainless steel tray, is placed in -40 DEG C of refrigerator freezings and is preserved.
4) by the liquid phase ingredient after high speed centrifugation, using circulating pump, circulating pump conveying pressure is set as 0.8MPa, is made Liquid phase ingredient removes solid phase impurity by 0.45 μm of miillpore filter.The efflux after micro porous filtration is collected, using circulating pump, Circulating pump conveying pressure is set as 1.5MPa, makes the feed liquid after removing solid phase impurity again by aperture for 1,000 dalton NF membrane carries out desalination and removing heavy metals ion, and concentrates the moisture of removing 70%, obtains 30% concentrate, percentage is each The quality of component accounts for the percentage of liquid phase ingredient gross mass.Concentrate carries out freeze-drying and removes moisture removal.First to lyophilized before lyophilized Machine carries out precooling, when freeze-drying machine partition board temperature is down to 4 DEG C, will fill the sea cucumber concentrate crossed in -40 DEG C of pre-freezes not Rust steel tray is placed on shelf, and it is -40 DEG C to set freeze-drying machine partition board temperature, opens water vessel refrigerating state, opens vavuum pump, if It is 15Pa to put vacuum, is persistently freezed 10 hours.Product after lyophilized crushes 30 minutes in pulverizer, crosses 60 mesh sieves, obtains water The sea cucumber extract of insoluble portion.
5) by the solid-phase component after high speed centrifugation, it is freeze-dried with the same operating method described in step 4) Remove moisture removal.Solid-phase component after lyophilized, is crushed 30 minutes in pulverizer, is crossed 60 mesh sieves, is obtained thick sea cucumber powder.Every 1 kilogram thick Sea cucumber powder is placed in 5L extractive reaction kettle, carries out CO2Supercritical extract, regulation and control extraction conditions are：Extraction kettle Stress control exists 25MPa, extraction kettle temperature control is at 40 DEG C, CO2Flow velocity is 25L/h, dynamic extraction 3 hours, obtains the sea of non-water-soluble fractions Conopsea extraction.
Embodiment 2 extracts active component from sea cucumber
1) fresh extra large stichopus japonicus (Stichopus japonicus) is taken, removes clean internal organ, removes residual in intestinal wall Silt, it is put into rinse bath, with clean running water cleaning three times, adds deionized water cleaning once, pull out and drain away the water, Obtain sea cucumber skin.Start meat grinder, sea cucumber is rubbed one by one, the sea cucumber skin after rubbing is collected in stainless steel cask, adds 4 times of weight Deionized water, be thoroughly mixed, form broken sea cucumber skin mixed liquor.Start colloid mill, setting fineness is 200 mesh, will be broken The feeding mouth that broken sea cucumber skin mixed liquor gradually puts into colloid mill is ground, and the feed liquid of outlet grinds one repeatedly with the same manner again It is secondary, always it is co-mulled and made into twice, the sea cucumber slurry after finally being ground.
2) the sea cucumber slurry prepared is inputted in vacuum ultrasonic reactor, is evacuated to 100Pa, control temperature in the kettle 10 DEG C, the power for adjusting ultrasonic transmitter is 500W, sets ultrasonic wave tranmitting frequency as 25KHz, pulse cycle frequency 8 times/ Second, the meter per second of acoustic beam 1700, liquid surface sound pressure level is 20 °, starts ultrasonic wave program, controls ultrasonic intensity, produces sea cucumber slurry Raw obvious vortex.Ultrasonication 200 minutes, sea cucumber histocyte wall is set fully to be crushed, sea cucumber active component is released Release, obtain sea cucumber active component extract.
3) by sea cucumber active component extract, in the centrifuge tube of addition supercentrifuge in batches, setting high-speed centrifugation The revolution of machine is 12,000rpm, and centrifugation time 30 minutes, temperature is set as 10 DEG C, starts centrifuge and starts centrifugal process.Centrifugation After end, careful collection centrifuged supernatant, i.e. liquid phase ingredient, it is placed in a clean stainless steel cask.Remaining solid-phase component It is collected in stainless steel tray, is placed in -40 DEG C of refrigerator freezings and preserves.
4) by the liquid phase ingredient after high speed centrifugation, using circulating pump, circulating pump conveying pressure is set as 0.8MPa, is made Liquid phase feed liquid removes solid phase impurity by 0.22 μm of miillpore filter.The efflux after micro porous filtration is collected, using circulating pump, Circulating pump conveying pressure is set as 1.5MPa, makes the feed liquid after removing solid phase impurity again by aperture for 1,000 dalton NF membrane carries out desalination and removing heavy metals ion, and concentrates the moisture of removing 70%, obtains 30% concentrate.Concentrate is entered Moisture removal is removed in row freeze-drying.Precooling first is carried out to freeze dryer before lyophilized, when freeze-drying machine partition board temperature is down to 4 DEG C, will be filled The stainless steel tray for the sea cucumber concentrate crossed in -40 DEG C of pre-freezes is placed on shelf, and it is -40 to set freeze-drying machine partition board temperature DEG C, water vessel refrigerating state is opened, opens vavuum pump, setting vacuum is 15Pa, is persistently freezed 8 hours.Product after lyophilized Crushed 20 minutes in pulverizer, cross 60 mesh sieves, obtain the sea cucumber extract of water-soluble portion.
5) by the solid-phase component after high speed centrifugation, it is freeze-dried with the same operating method described in step 4) Remove moisture removal.Solid-phase component after lyophilized, is crushed 30 minutes in pulverizer, is crossed 60 mesh sieves, is obtained thick sea cucumber powder.Every 1 kilogram thick Sea cucumber powder is placed in 5L extractive reaction kettle, carries out CO2Supercritical extract, regulation and control extraction conditions are：Extraction kettle Stress control exists 25MPa, extraction kettle temperature control is at 35 DEG C, CO2Flow velocity is 30L/h, dynamic extraction 2.5 hours, obtains non-water-soluble fractions Sea cucumber extract.
Embodiment 3 extracts active component from sea cucumber
1) fresh extra large stichopus japonicus (Stichopus japonicus) is taken, removes clean internal organ, removes residual in intestinal wall Silt, it is put into rinse bath, with clean running water cleaning three times, adds deionized water cleaning once, pull out and drain away the water. Start meat grinder, sea cucumber skin rubbed one by one, the sea cucumber skin after rubbing is collected in stainless steel cask, add 4 times of weight go from Sub- water, is thoroughly mixed, and forms broken sea cucumber skin mixed liquor.Start colloid mill, setting fineness is 200 mesh, by broken sea cucumber The feeding mouth that skin mixed liquor gradually puts into colloid mill is ground, and the feed liquid of outlet is ground once repeatedly with the same manner again, always It is co-mulled and made into twice, the sea cucumber slurry after finally being crushed.
2) the sea cucumber slurry prepared is inputted in vacuum ultrasonic reactor, is evacuated to 100Pa, control temperature in the kettle 0 DEG C, the power for adjusting ultrasonic transmitter is 500W, sets ultrasonic wave tranmitting frequency as 25KHz, pulse cycle frequency 15 times/ Second, the meter per second of acoustic beam 1700, liquid surface sound pressure level is 20 °, starts ultrasonic wave program, controls ultrasonic intensity, produces sea cucumber slurry Raw obvious vortex.Ultrasonication 150 minutes, sea cucumber histocyte wall is set fully to be crushed, sea cucumber active component is released Release, obtain sea cucumber active component extract.
3) by sea cucumber active component extract, in the centrifuge tube of addition supercentrifuge in batches, setting high-speed centrifugation The revolution of machine is 12,000rpm, and centrifugation time 60 minutes, temperature is set as 0 DEG C, starts centrifuge and starts centrifugal process.Centrifugation After end, careful collection centrifuged supernatant, i.e. liquid phase ingredient, it is placed in a clean stainless steel cask.Remaining solid-phase component It is collected in stainless steel tray, is placed in -40 DEG C of refrigerator freezings and preserves.
4) by the liquid phase ingredient after high speed centrifugation, using circulating pump, circulating pump conveying pressure is set as 0.8MPa, is made Liquid phase ingredient removes solid phase impurity by 0.80 μm of miillpore filter.The efflux after micro porous filtration is collected, using circulating pump, Circulating pump conveying pressure is set as 1.5MPa, makes the feed liquid after removing solid phase impurity again by aperture for 1,000 dalton NF membrane carries out desalination and removing heavy metals ion, and concentrates the moisture of removing 70%, obtains 30% concentrate.Concentrate is carried out Moisture removal is removed in freeze-drying.Precooling first is carried out to freeze dryer before lyophilized, when freeze-drying machine partition board temperature is down to 4 DEG C, will be filled The stainless steel tray of sea cucumber concentrate through being crossed in -40 DEG C of pre-freezes is placed on shelf, and it is -40 DEG C to set freeze-drying machine partition board temperature, Water vessel refrigerating state is opened, opens vavuum pump, setting vacuum is 15Pa, is persistently freezed 12 hours.Product after lyophilized in Crushed 45 minutes in pulverizer, cross 60 mesh sieves, obtain the sea cucumber extract of water-soluble portion.
5) by the solid-phase component after high speed centrifugation, it is freeze-dried with the same operating method described in step 4) Remove moisture removal.Solid-phase component after lyophilized, is crushed 45 minutes in pulverizer, is crossed 60 mesh sieves, is obtained thick sea cucumber powder.Every 1 kilogram thick Sea cucumber powder is placed in 5L extractive reaction kettle, carries out CO2Supercritical extract reacts, and regulation and control extraction conditions is：Extraction kettle pressure control System is in 25MPa, and extraction kettle temperature control is at 45 DEG C, CO2Flow velocity is 20L/h, dynamic extraction 4 hours, obtains non-water-soluble fractions Sea cucumber extract.
Comparative example 1 commonly extracts the processing mode of active material of sea cucumber
1) fresh extra large stichopus japonicus (Stichopus japonicus) is taken, removes clean internal organ, removes residual in intestinal wall Silt, it is put into rinse bath, with clean running water cleaning three times, adds deionized water cleaning once, pull out and drain away the water, Obtain sea cucumber skin.Start meat grinder, sea cucumber skin is rubbed one by one, the sea cucumber after rubbing is collected in stainless steel cask, adds 4 times of weight Deionized water, be thoroughly mixed, form broken sea cucumber skin mixed liquor.Start colloid mill, setting fineness is 200 mesh, will be broken The feeding mouth that broken sea cucumber skin mixed liquor gradually puts into colloid mill is ground, and the feed liquid of outlet grinds one repeatedly with the same manner again It is secondary, always it is co-mulled and made into twice, the sea cucumber slurry after finally being ground.
2) the sea cucumber slurry after milled processed is imported into filter press and carries out press filtration, collect respectively solid-phase component and liquid phase into Point.Liquid phase ingredient is placed in a clean stainless steel cask.Solid-phase component is collected in stainless steel tray.
3) by the liquid phase ingredient after press filtration, add in rotary evaporator, set 40 DEG C of evaporating temperature, vacuumize rotation and steam Hair is until dry.Dried product crushes 30 minutes in pulverizer, crosses 60 mesh sieves, obtains the sea cucumber extraction of water-soluble portion Thing.
4) the stainless steel tray of solid-phase component will be filled, is placed in cold wind dryer, sets 40 DEG C of wind-warm syndrome, cold wind is opened and dries Dry machine, persistently dry 10 hours, obtain the sea cucumber extract of non-water-soluble fractions.
Comparative example 2 commonly extracts the processing mode of active material of sea cucumber.
1) fresh extra large stichopus japonicus (Stichopus japonicus) is taken, removes clean internal organ, removes residual in intestinal wall Silt, it is put into rinse bath, with clean running water cleaning three times, adds deionized water cleaning once, pull out and drain away the water, Obtain sea cucumber skin.Start meat grinder, sea cucumber skin is rubbed one by one, the sea cucumber skin after rubbing is collected in stainless steel cask, adds 4 times of weights The deionized water of amount, is thoroughly mixed, and forms broken sea cucumber skin mixed liquor.Start colloid mill, setting fineness is 200 mesh, will The feeding mouth that broken sea cucumber skin mixed liquor gradually puts into colloid mill is ground, and the feed liquid of outlet is ground repeatedly with the same manner again Once, always it is co-mulled and made into twice, the sea cucumber slurry after finally being ground.
2) by the sea cucumber slurry after milled processed, adding in the centrifuge tube of centrifuge in batches, turning for centrifuge is set Number is 6,000rpm, at room temperature centrifugation time 60 minutes.Centrifugation terminate after, careful collection centrifuged supernatant, obtain liquid phase into Point, it is placed in a clean stainless steel cask.Remaining solid-phase component is collected in stainless steel tray.
3) by the liquid phase ingredient after centrifugation, add in rotary evaporator, set 60 DEG C of evaporating temperature, vacuumize rotation and steam Hair is until dry.Dried product crushes 30 minutes in pulverizer, crosses 60 mesh sieves, obtains sea cucumber powder, obtain water-soluble portion Sea cucumber extract.
4) the stainless steel tray of solid-phase component will be filled, is placed in cold wind dryer, sets 80 DEG C of wind-warm syndrome, cold wind is opened and dries Dry machine, persistently dry 10 hours, obtain the sea cucumber extract of non-water-soluble fractions.
The beche-de-mer products that embodiment and comparative example are prepared, detection, as a result as shown in table 2.Using embodiment 1, in fact Apply in the sea cucumber extract that example 2 and the method for embodiment 3 obtain, its total protein content, DHA content, EPA content significantly greater than with The sea cucumber extract that comparative example 1 and the method for comparative example 2 obtain；And content of beary metal is then substantially less than using comparative example 1 and contrast The sea cucumber extract that the method for example 2 obtains.
The embodiment of table 2. and comparative example method sea cucumber extraction product detection result

Claims (10)

  1. A kind of 1. method that active component is extracted from sea cucumber, it is characterised in that it comprises the following steps：Sea cucumber slurry is passed through Ultrasonication, produce sea cucumber active component extract；The temperature of the ultrasonication is 0 DEG C -10 DEG C.
  2. 2. the method as described in claim 1, it is characterised in that：Described sea cucumber slurry is obtained by sea cucumber after pretreatment Arrive；
    And/or described ultrasonication is carried out in vacuum ultrasonic reactor；
    And/or the treatment temperature in described ultrasonication is 4 DEG C；
    And/or the time of described ultrasonication is 150-200 minutes, preferably 180 minutes；
    And/or power is 300-700W, preferably 500W in described ultrasonication；
    And/or tranmitting frequency is 20-30KHz, preferably 25KHz in described ultrasonication；
    And/or pulse cycle frequency is 5-15 times/second, preferably 10 times/second in described ultrasonication；
    And/or acoustic beam is 1200-2200 meter per seconds, preferably 1700 meter per seconds in described ultrasonication；
    And/or liquid surface sound pressure level is 15-55 °, preferably 55 ° in described ultrasonication；
    And/or vacuum is 80-120Pa, preferably 100Pa in described ultrasonication；
    And/or described sea cucumber active component extract stores at -30 to -50 DEG C, preferably -40 DEG C.
  3. 3. method as claimed in claim 2, it is characterised in that：Described pretreatment comprises the steps：
    (a) by sea cucumber impurity elimination, sea cucumber skin is obtained；
    (b) described sea cucumber skin is crushed, the sea cucumber skin crushed；
    (c) the sea cucumber skin of described crushing is ground together with water, obtains described sea cucumber slurry；
    In described step (a), described sea cucumber is preferably extra large stichopus japonicus (Stichopus japonicus)；Described impurity elimination is excellent Elect as and remove the internal organs and silt；Also include the operation with water rinse after described impurity elimination；The operation of described water rinse is preferably wrapped Include the following steps：Cleaned with clean running water, then cleaned with deionized water or distilled water；The described operation knot with water rinse Also include draining operation after beam；
    In described step (b), described crushing is preferably to be crushed using meat grinder；
    In described step (c), the mass values of described water and the sea cucumber skin of described crushing are preferably 3~5；Described grinds Mill is preferably using high speed homogenizer grinding or colloid mill grinding；The number of described grinding is preferably 1 time or 2 times.
  4. A kind of 4. sea cucumber active component extract, it is characterised in that its method system described according to claim any one of 1-3 , and it is freeze-dried after obtain；
    Wherein, the temperature of described freeze-drying is preferably -30 to -50 DEG C；More preferably -40 DEG C；Described freeze-drying Vacuum is preferably 10-20Pa, more preferably 15Pa；The time of described freeze-drying is preferably 8-12 hours, more preferably 10 hours；Described freeze-drying is carried out preferably in freeze drier；It is excellent before the operation of described freeze-drying is carried out Described freeze drier is first carried out pre-freeze by choosing.
  5. A kind of 5. method that active component is extracted from sea cucumber, it is characterised in that it comprises the following steps：Will be such as claim 1- Described sea cucumber active component extract made from method described in 3 any one through being centrifuged at a high speed to obtain solid-phase component and Liquid phase ingredient.
  6. 6. method as claimed in claim 5, it is characterised in that the method that active component is extracted from sea cucumber also includes The operation handled described liquid phase ingredient；The processing of described liquid phase ingredient preferably includes the following steps：Will be described The operation that liquid phase ingredient passes through filtering with microporous membrane；Wherein, described miillpore filter aperture is preferably 0.22~0.8 μm；It is described Filtering with microporous membrane when pressure be preferably 0.5-1.0MPa；Also include passing through NF membrane after described filtering with microporous membrane The operation of nanofiltration；The pressure of described nanofiltration is preferably 1.0-1.8MPa；The aperture of described NF membrane is preferably 500-1,500 Dalton；The processing of described liquid phase ingredient still further comprises the following steps：It is freeze-dried after described nanofiltration；Institute The temperature for the freeze-drying stated is preferably -30 to -50 DEG C；The vacuum of described freeze-drying is preferably 10-20Pa；Described The time of freeze-drying is preferably 8-12 hours；The processing of described liquid phase ingredient may further include described freeze-drying After crushed, sieve, that is, obtain the sea cucumber extract of water-soluble portion.
  7. 7. method as claimed in claim 5, it is characterised in that the method that active component is extracted from sea cucumber also includes The operation handled described solid-phase component；The processing of described solid-phase component preferably includes the following steps：Will be described Solid-phase component, carry out CO2Supercritical extract, produce the sea cucumber composition of non-water-soluble fractions；Further preferably comprise the following steps： By described solid-phase component after freeze-drying and crushing, thick sea cucumber powder is obtained, then carries out CO again2Supercritical extract；Wherein, Described freeze-drying and the condition of crushing are the same as liquid phase ingredient lyophilized dry in claim 6 and the condition crushed；Described CO2 The pressure of supercritical extract is preferably 20-30MPa；Described CO2The temperature of supercritical extract is preferably 30-50 DEG C；Described CO2CO in supercritical extract2Flow velocity is preferably 20-30L/h；Described CO2The time of supercritical extract is preferably 2-3 hours.
  8. 8. a kind of sea cucumber active component extract, it is made as the method described in claim any one of 5-7.
  9. 9. a kind of sea cucumber extract, it is characterised in that it is made by following methods, and described method comprises the following steps：Will be as Described sea cucumber active component extract is by being centrifuged at a high speed to obtain made from method described in claim any one of 1-3 Liquid phase ingredient；Described liquid phase ingredient is also handled using the processing method of the liquid phase ingredient described in claim 6.
  10. 10. a kind of sea cucumber extract, it is characterised in that it is made by following methods, and described method comprises the following steps：Will be as Described sea cucumber active component extract is by being centrifuged at a high speed to obtain made from method described in claim any one of 1-3 Solid-phase component, described solid-phase component are handled using the processing method of the procedure component described in claim 7.
CN201710677091.6A 2017-08-09 2017-08-09 A kind of method and its extract that active component is extracted from sea cucumber Pending CN107334096A (en)

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CN107955081A (en) * 2017-11-20 2018-04-24 黑龙江美丽石岛科技开发有限公司 The combined extraction method of a variety of nutriments in a kind of sea cucumber
CN108030004A (en) * 2017-12-05 2018-05-15 河北汉唯生物制药有限公司 A kind of sea cucumber nutrient composition, its preparation method and the food containing it
CN108384726A (en) * 2018-04-25 2018-08-10 杭州中美华东制药有限公司 Aweto mycelium and its fermentation medium and fermentation process
CN109452578A (en) * 2018-12-29 2019-03-12 海南椰岛酒业发展有限公司 A kind of preparation method of marine animal extract
CN109620853A (en) * 2017-10-09 2019-04-16 董玲 A kind of extraction process of medicinal material
CN111808673A (en) * 2020-07-09 2020-10-23 江苏海宏制药有限公司 Subcritical fluid extraction process of sea cucumber oil

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CN103005528A (en) * 2007-07-08 2013-04-03 谭攸恒 Preparation method of sea cucumber extract

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CN103005528A (en) * 2007-07-08 2013-04-03 谭攸恒 Preparation method of sea cucumber extract
CN101411523A (en) * 2008-11-14 2009-04-22 钱武刚 Method for extracting original liquid of trepang by ultrasonic wave

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109620853A (en) * 2017-10-09 2019-04-16 董玲 A kind of extraction process of medicinal material
CN107955081A (en) * 2017-11-20 2018-04-24 黑龙江美丽石岛科技开发有限公司 The combined extraction method of a variety of nutriments in a kind of sea cucumber
CN108030004A (en) * 2017-12-05 2018-05-15 河北汉唯生物制药有限公司 A kind of sea cucumber nutrient composition, its preparation method and the food containing it
CN108384726A (en) * 2018-04-25 2018-08-10 杭州中美华东制药有限公司 Aweto mycelium and its fermentation medium and fermentation process
CN109452578A (en) * 2018-12-29 2019-03-12 海南椰岛酒业发展有限公司 A kind of preparation method of marine animal extract
CN111808673A (en) * 2020-07-09 2020-10-23 江苏海宏制药有限公司 Subcritical fluid extraction process of sea cucumber oil

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