CN101857623B - Method for extracting GP4G from organism - Google Patents

Method for extracting GP4G from organism Download PDF

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CN101857623B
CN101857623B CN2010101819568A CN201010181956A CN101857623B CN 101857623 B CN101857623 B CN 101857623B CN 2010101819568 A CN2010101819568 A CN 2010101819568A CN 201010181956 A CN201010181956 A CN 201010181956A CN 101857623 B CN101857623 B CN 101857623B
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gp4g
extraction
carry out
solution
adopt
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CN101857623A (en
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郑春阳
魏国祥
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TIANJIN QIANGWEITE BIO-TECH Co Ltd
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TIANJIN QIANGWEITE BIO-TECH Co Ltd
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Abstract

The invention provides a method for extracting GP4G (polyphosphoric guanosine) from organism, which adopts the following steps: pretreatment of raw material, dehydration, breaking, leaching, neutralization, adsorption, elution, desalination and vacuum concentration. With the technical scheme, the method has the advantages of high yield, low production cost and industrialized production.

Description

A kind of method of from organism, extracting GP4G
Technical field
The present invention relates to a kind of method of extracting active substance in the organism, particularly a kind of method of from organism, extracting GP4G (four di(2-ethylhexyl)phosphate guanosines).
Background technology
GP4G (P 1, P 4-Di (guanosine 5 ') tetraphosphate), chemistry four di(2-ethylhexyl)phosphate guanosines by name, it is plain to be called as cell activation regeneration in cosmetic circle, porous has Cucumber Extract to skin corium, helps to repair damaged cell, strengthen the refresh function of cell, make the nutrient mediation efficient between the cell higher; Promote the collagen protein composition synthetic, promote the cell overall activity, delay wrinkle and produce, improve and the desalination wrinkle, help skin to recover to compact and elasticity.Has vast market prospect at cosmetic industry.
There is the extracting method about GP4G of report to see the article The occurrence of P that F.J.FINAMORE and A.H.WARNER delivered at The Journal of Biological Chemistry in 1963 at present 1, P 4-Diguanosine 5 '-Tetraphosphate inBrine shrimp eggs.But according to the method for the extraction GP4G of this bibliographical information, the yield of its GP4G is low, equipment requirements condition height, and the production cost height is unfavorable for suitability for industrialized production.
Summary of the invention
Given this, the purpose of this invention is to provide a kind of method of from organism, extracting GP4G, compare, have the yield height, production cost is low, helps the characteristics of suitability for industrialized production with method in the document.
The present invention adopts following technical scheme: as starting material, carry out the operation of following steps with the biomaterial that contains GP4G:
Raw materials pretreatment: take by weighing the biological starting material of 1000kg~2000kg, screen, remove foreign material, clean again with 100~200 purpose screen clothes;
Dehydration: the biological starting material after will handling are placed on the conveyer that is provided with 100~200 eye mesh screens and dewater, and transmit with the speed of 0.5m/min~2m/min;
Broken: the biological starting material after will dewatering carry out fragmentation;
Leaching: the mixed with-10 ℃~20 ℃ the acidic solution of the biological raw-material pure dry weight after the fragmentation and 1mol/L~3mol/L is pressed 1: 1~10kg: L, fully stir 30min~120min, separation of supernatant;
Neutralization: behind 1~10 times of the vacuum concentration, with supernatant liquor pH to 7~8 after 2mol/L~5mol/L basic solution adjustment leaching, collect the solid precipitate, remove clear liquid, the solid precipitate is adopted distilled water or deionized water dissolving by 5%~30% concentration, adopt 2mol/L~5mol/L acidic solution to transfer clear liquid pH to 2~5 again, prepare upper prop;
Absorption: above-mentioned solution adsorbs with storng-acid cation exchange resin, adsorb saturated after, injection de-salted water or deionized water carry out binder;
Wash-out: the acid eluent with 0.5mol/L~2mol/L carries out the wash-out desorb, and the Fractional Collections target product;
Desalination: adopt weak base anion-exchange resin to carry out overcurrent technology decolouring desalting treatment then;
Vacuum concentration: at working pressure be-0.085MPa~-0.092MPa, temperature is under 30 ℃~60 ℃ the vacuum state, when the content of GP4G in the solution reaches 6%, to begin to collect GP4G.
The biomaterial of the described GP4G of containing be contain high salt and contain high mineral or halophilic marine plankton and interior lake in shell class biology.Be preferably and contain high salt and contain high mineral or halophilic halogen worm or artemia cysts.
The acidic solution of using in the described leaching step is HCl, HClO 4, H 2SO 4, HNO 3, HCOOH, CH 3Among the COOH one or more.
The separation method of supernatant liquor is siphon, centrifugal, one or more modes in filtering in the described leaching step.
Used basic solution is NaOH in the described neutralization procedure, KOH, one or more in the ammoniacal liquor.
Acid eluent in the described elution step is formic acid, acetate, HCl, H 2SO 4In one or more.
The GP4G product of described final acquisition is liquid or solid-state, is preferably liquid state.
In actually operating, often add NaCl, (NH in the acidic solution of using in the leaching process 4) 2SO 4, salts such as KCl are used for the effect strengthening leaching; After for the first time the supernatant liquor leaching being finished, also have some solid insoluble residues, these solid insolubles and acidic solution are carried out secondary, three leachings in 1: 1~10 ratio, twice leaching liquid in back can be successively as the first time of next batch, leaching liquid uses for the second time, Cao Zuo benefit can reduce solvent load and wastewater flow rate like this, can improve the lixiviate yield simultaneously and save extraction time.With remaining solid insoluble flame filter press, working pressure is that 10MPa~20MPa carries out filtration treatment again, collects supernatant liquor, and filter residue is used for other purposes.
After obtaining the GP4G of vacuum concentration, can make following process respectively by the different specification requirement of the finished product.
The method of mentioning in the present invention and the document is compared, and has the following advantages: (1) the present invention does not adopt and directly transfers the sedimentary method of pH, afterwards transfers pH but take to concentrate earlier, has improved yield so greatly, and the while has also been saved the consumption of alkali; (2) when adopting cation seperation column absorption, be after absorption is saturated, just to carry out the setting-out wash-out, increased the resin absorption amount like this, improved service efficiency, reduced cost; (3) be to adopt weak base anion-exchange resin to carry out overcurrent technology decolouring desalting treatment during desalination, rather than the charcoal of mentioning in document absorption or dialysis desalination, can play the effect of desalination and decolouring so simultaneously, improved efficient.In addition, also adopt Plate Filtration to substitute the technology of high speed centrifugation and B in actually operating, operational condition requires low, is beneficial to industrialization, large-scale production.In sum, the present invention has the yield height, production cost is low, is beneficial to the characteristics of suitability for industrialized production.
Embodiment
Describe in conjunction with specific embodiments.
Embodiment 1
Carry out the extraction of GP4G by following steps:
The first step: take by weighing 1000kg salt solution good year worm's ovum, moto-vibro screen vibration with 100 eye mesh screens, remove dust and tiny solid impurity, artemia cysts and bigger solid impurity are stayed on the sieve, enter the uniflux service sink then, utilize the proportion difference, adopt 120 purpose screen clothes to fish for the artemia cysts of keeping afloat at the pond the other end;
Second step: the salt solution good year worm's ovum of fishing for is placed on the belt conveyer at 120 eye mesh screens, 30 ° of inclination angles, speed with 2m/min transmits, the moisture content that artemia cysts is had leaks down by screen cloth, and the other end of conveyer links to each other with the pulverizer opening for feed, makes the artemia cysts of drip-dry water directly enter pulverizer;
The 3rd step: by pulverizer worm's ovum being crushed to its granular size can be good by 140 purpose screen clothes;
The 4th step: the pure dry weight of the salt solution good year worm's ovum after the fragmentation and-4 ℃ the acidic solution HCl of 1mol/L are pressed 1: the mixed of 1kg: L, fully stir 30min, adopt centrifugation to separate supernatant;
The 5th step: behind 2 times of the vacuum concentration,, collect the solid precipitate with supernatant liquor pH to 7~8 after the 2mol/L basic solution adjustment leaching, remove clear liquid, the solid precipitate is pressed 30% concentration with distilled water or deionized water dissolving, transfer clear liquid pH to 2~5 with the 2mol/L acidic solution again, prepare upper prop;
The 6th step: the solution after the above-mentioned neutralization carries out upper prop absorption with 0.5BV/h speed at 732 storng-acid cation exchange resins, adsorb saturated after, stop upper prop, adopt de-salted water to carry out binder then, the hydraulic fluid color is thin out to be stopped, binder water is gone here and there post and is adsorbed;
The 7th step: carry out the wash-out desorb, the Fractional Collections target product with the acid eluent of the HCl of 0.5mol/L;
The 8th step: upper prop speed is 0.5BV/h, adopts weak base anion-exchange resin to carry out overcurrent technology decolouring desalting treatment; To collecting pH value of solution is to stop desalting treatment at 6.5 o'clock;
The 9th step: be-0.090MPa that temperature is under 40 ℃ the vacuum state, when the content of GP4G in the solution reaches 6%, to begin to collect GP4G at vacuum operating pressure.
Adopt the method in the document to extract with 1000kg salt solution good year worm's ovum, the yield of GP4G among the GP4G yield of gained and the present invention compared, see the following form:
Method Yield
Literature method 10% (solid-state)
The present invention 78% (liquid state)
Embodiment 2
Carry out the extraction of GP4G by following steps:
The first step: the Artemia Salina worm's ovum that takes by weighing 1500kg, moto-vibro screen vibration with 100 eye mesh screens, remove dust and tiny solid impurity, artemia cysts and bigger solid impurity are stayed on the sieve, enter the uniflux service sink then, utilize the proportion difference, adopt 120 purpose screen clothes to fish for the artemia cysts of keeping afloat at the pond the other end;
Second step: the artemia cysts of fishing for is placed on the belt conveyer at 120 eye mesh screens, 30 ° of inclination angles, speed with 1m/min transmits, the moisture content that artemia cysts is had leaks down by screen cloth, and the other end of conveyer links to each other with the pulverizer opening for feed, makes the artemia cysts of drip-dry moisture directly enter pulverizer;
The 3rd step: by pulverizer worm's ovum being crushed to its granular size can be good by 140 purpose screen clothes;
The 4th step: the pure dry weight of the artemia cysts after the fragmentation and 0 ℃ the acidic solution HCl of 1.5mol/L are pressed 1: the mixed of 3kg: L, fully stir 70min, adopt the mode of siphon to separate supernatant;
The 5th step: behind 4 times of the vacuum concentration,, collect the solid precipitate with supernatant liquor pH to 7~8 after the 3mol/L basic solution adjustment leaching, remove clear liquid, the solid precipitate is pressed 25% concentration with distilled water or deionized water dissolving, transfer clear liquid pH to 2~5 with the 2mol/L acidic solution again, prepare upper prop;
The 6th step: the solution after the above-mentioned neutralization carries out upper prop absorption with 1BV/h speed at 732 storng-acid cation exchange resins, adsorb saturated after, stop upper prop, adopt ionized water to carry out binder then, the hydraulic fluid color is thin out to be stopped, binder water is gone here and there post and is adsorbed;
The 7th step: with the H of 1mol/L 2SO 4Acid eluent carries out the wash-out desorb, the Fractional Collections target product;
The 8th step: upper prop speed is 1BV/h, and adopting weak base anion-exchange resin to carry out overcurrent technology decolouring desalting treatment is to stop desalination at 7 o'clock to collecting pH value of solution;
The 9th step: be-0.092MPa that temperature is under 50 ℃ the vacuum state, when the content of GP4G in the solution reaches 6%, to begin to collect GP4G at vacuum operating pressure.
Adopt the method in the document to extract with the Artemia Salina worm's ovum of 1500kg, the yield of GP4G among the GP4G yield of gained and the present invention compared, see the following form:
Method Yield
The paper method 10% (solid-state)
This technology 75% (liquid state)
Embodiment 3
Carry out the extraction of GP4G by following steps, carry out solid crystal then:
The first step: take by weighing the 2000kg artemia cysts, moto-vibro screen vibration with 100 eye mesh screens, remove dust and tiny solid impurity, artemia cysts and bigger solid impurity are stayed on the sieve, enter the uniflux service sink then, utilize the proportion difference, adopt 120 purpose screen clothes to fish for the artemia cysts of keeping afloat at the pond the other end;
Second step: the artemia cysts of fishing for is placed on the belt conveyer at 120 eye mesh screens, 30 ° of inclination angles, speed with 2m/min transmits, the water that artemia cysts is had leaks down by screen cloth, and the other end of conveyer links to each other with the pulverizer opening for feed, makes the artemia cysts of drip-dry water directly enter pulverizer;
The 3rd step: by pulverizer worm's ovum being crushed to its granular size can be good by 140 purpose screen clothes;
The 4th step: with 15 ℃ the acidic solution HCOOH of the pure dry weight of the artemia cysts after the fragmentation and 3mol/L by 1: 10 (kg: mixed L), fully stir 120min, the mode of employing siphon is separated supernatant;
The 5th step: behind 5 times of the vacuum concentration,, collect the solid precipitate with supernatant liquor pH to 7~8 after the 4mol/L basic solution adjustment leaching, remove clear liquid, the solid precipitate is pressed 30% concentration with distilled water or deionized water dissolving, transfer clear liquid pH to 2~5 with the 2mol/L acidic solution again, prepare upper prop;
The 6th step: above-mentioned solution carries out upper prop absorption with 2BV/h speed at 732 storng-acid cation exchange resins, adsorb saturated after, stop upper prop, adopt ionized water to carry out binder then, the hydraulic fluid color is thin out to be stopped, binder water is gone here and there post and is adsorbed;
The 7th step: with the H of 1mol/L 2SO 4Acid eluent carries out the wash-out desorb, the Fractional Collections target product;
The 8th step: upper prop speed is 2BV/h, and adopting weak base anion-exchange resin to carry out overcurrent technology decolouring desalting treatment is 7 to stop to collecting pH value of solution;
The 9th step: be-0.085MPa that temperature is under 60 ℃ the vacuum state, when the content of GP4G in the solution reaches 6%, to begin to collect GP4G at working pressure;
The tenth step: adopt 95% ethanol, 1: 3 by volume (V/V (ethanol)) slowly adds the alcohol crystallization while stirring to concentrated solution, allows crystal separate out naturally.-4 ℃ low incubation crystalline substances 2~10 hours;
The 11 step: it is centrifugal to adopt centrifugal speed 4000rpm to carry out, and collects crystal, and mother liquor returns the 5th step operation;
The 12 step: dry employing small size vacuum double-cone dryer, 50 ℃ of temperature, vacuum operating pressure-0.092MPa, moisture≤1% o'clock discharging;
The 13 step:, pack after the assay was approved through 60 purpose boltings.
Adopt the method in the document to extract with the 2000kg artemia cysts, the yield of GP4G among the GP4G yield of gained and the present invention compared, see the following form:
Method Yield
The paper method 10% (solid-state)
This technology 50% (solid-state)
The above is the preferred embodiments of the present invention only, is not limited to the present invention, and for a person skilled in the art, the present invention can have various changes and variation.Within the spirit and principles in the present invention all, any modification of being done, be equal to replacement, improvement etc., all should be included within protection scope of the present invention.

Claims (7)

1. method of extracting GP4G from organism is characterized in that: adopt the biomaterial that contains GP4G as starting material, carry out the operation of following steps:
Raw materials pretreatment: take by weighing the biological starting material of 1000kg~2000kg, screen, remove foreign material, clean again with 100~200 eye mesh screens;
Dehydration: the biological starting material after will handling are placed on the conveyer that is provided with 100~200 eye mesh screens and dewater, and transmit with the speed of 0.5m/min~2m/min;
Broken: the biological starting material after will dewatering carry out fragmentation;
Leaching: the mixed with-10~20 ℃ the acidic solution of the biological raw-material pure dry weight after the fragmentation and 1mol/L~3mol/L is pressed 1: 1~10kg: L, fully stir 30min~120min, separation of supernatant;
Neutralization: behind 1~10 times of the vacuum concentration, adopt supernatant liquor pH to 7~8 after 2mol/L~5mol/L basic solution adjustment is leached, collect the solid precipitate, remove clear liquid, with the solid precipitate by 5%~30% concentration with distilled water or deionized water dissolving, transfer clear liquid pH to 2~5 with 1~5mol/L acidic solution again, prepare upper prop;
Absorption: solution adsorbs with storng-acid cation exchange resin, adsorb saturated after, injection de-salted water or deionized water carry out binder;
Wash-out: the acid eluent with 0.5mol/L~2mol/L carries out the wash-out desorb, and the Fractional Collections target product;
Desalination: adopt weak base anion-exchange resin to carry out overcurrent technology decolouring desalting treatment then;
Vacuum concentration: at vacuum operating pressure be-0.085MPa~-0.092MPa, temperature is that 30 ℃~60 ℃ vacuum state concentrates down, when the content of GP4G in the solution reaches 6%, begins to collect GP4G;
The biomaterial of the described GP4G of containing is for containing high salt and containing high mineral or halophilic artemia cysts.
2. the method for extraction GP4G according to claim 1, it is characterized in that: the acidic solution of using in the described leaching step is HCl, HClO 4, H 2SO 4, HNO 3, HCOOH, CH 3Among the COOH one or more.
3. the method for extraction according to claim 1 GP4G is characterized in that: the separation method of supernatant liquor is siphon, centrifugal, one or more modes in filtering in the described leaching step.
4. the method for extraction GP4G according to claim 1, it is characterized in that: used basic solution is NaOH in the described neutralization procedure, KOH, one or more in the ammoniacal liquor.
5. the method for extraction GP4G according to claim 1, it is characterized in that: the acid eluent in the described elution step is formic acid, acetate, HCl, H 2SO 4In one or more.
6. the method for extraction GP4G according to claim 1 is characterized in that: the final GP4G product that obtains is for liquid or solid-state.
7. the method for extraction GP4G according to claim 6 is characterized in that: the final GP4G product that obtains is for liquid.
CN2010101819568A 2010-05-25 2010-05-25 Method for extracting GP4G from organism Active CN101857623B (en)

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Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101999670A (en) * 2010-10-22 2011-04-06 天津强微特生物科技有限公司 Regenerating food capable of alleviating fatigue and rejuvenating and preparation method thereof
CN104974207B (en) * 2014-04-03 2017-07-28 珀莱雅化妆品股份有限公司 A kind of method that GP4G is extracted in the artemia cysts from salt water
ES2650981B2 (en) * 2016-07-20 2018-05-04 Ocupharm Diagnostics, Sl Preparation and use of an extract of Artemia saline to treat the ocular surface

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1304305A (en) * 1998-01-29 2001-07-18 科蒂股份有限公司 Cosmetic product based on artemia salina extracts for regenerating anmd stimulating skin cells

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2004238297A (en) * 2003-02-04 2004-08-26 Arysta Lifescience Corp Skin stress-resistant (stress-protecting) cosmetic

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1304305A (en) * 1998-01-29 2001-07-18 科蒂股份有限公司 Cosmetic product based on artemia salina extracts for regenerating anmd stimulating skin cells

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
F. J. FINAMORE, et al..The Occurrence of P1, P4-Diguanosine 5’-Tetraphosphate in Brine Shrimp Eggs.《THE JOURNAL OF BIOLOGICAL CHEMISTRY》.1963,第238卷(第1期),344-348. *
F.J.FINAMORE et al..The Occurrence of P1
JP特开2004-238297A 2004.08.26

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Denomination of invention: Method for extracting GP4G from organism

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