CN113563490B - Carrageenan extract and preparation method and application thereof - Google Patents

Carrageenan extract and preparation method and application thereof Download PDF

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CN113563490B
CN113563490B CN202110989471.XA CN202110989471A CN113563490B CN 113563490 B CN113563490 B CN 113563490B CN 202110989471 A CN202110989471 A CN 202110989471A CN 113563490 B CN113563490 B CN 113563490B
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carrageenan
carrageenan extract
ethanol
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CN113563490A (en
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孙云起
裴运林
刘涵
郭朝万
胡露
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Guangdong Marubi Biological Technology Co Ltd
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    • C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract

The invention relates to a carrageenan extract and a preparation method and application thereof. The preparation method of the carrageenan extract comprises the steps of freeze drying, reflux degreasing, concentration, centrifugation, addition of a solvent to remove algin, ethanol precipitation and the like, the preparation method is simple to operate, and the prepared carrageenan extract has a remarkable antioxidant effect and can be applied to the field of food additives or medicine additives.

Description

Carrageenan extract and preparation method and application thereof
Technical Field
The invention belongs to the technical field of polysaccharide, relates to an algae polysaccharide extract and a preparation method and application thereof, and particularly relates to a carrageenan extract and a preparation method and application thereof.
Background
With the development of medical technology, researchers have found that the causes and development of tumors, inflammations, aging, and the like are closely related to radical and lipid peroxidation, and thus, there is an increasing interest in finding substances capable of inhibiting radical and lipid peroxidation. In particular, the search for natural, non-toxic antioxidants has been an important research direction for pharmaceutical and food additives. With the concern of the biological activity of marine algae, research on algae extracts has been greatly advanced, and research shows that algae extracts have significant antioxidant activity, for example, red algae extracts not only have a certain inhibitory effect on lipid peroxidation induced by AAPH, but also have a significant inhibitory effect on DPPH.
Carrageenan is a low-fat, high-protein macroalgae belonging to the family Gigartinaceae, the phylum Rhodophyta, the genus Carrageenan, and is one of the important algae for carrageenan production. The carrageenin is rich in active substances, mainly comprises seaweed pigment, fatty acid, amino acid, sterol, polysaccharide, vitamin, etc., and has good bioactivity.
Polysaccharide substances are a class of biomacromolecules ubiquitous in biological organisms and are important components of cytoskeletons and various endogenous bioactive molecules. The polysaccharide can be used for regulating the physiological function of organisms in a free form or in a form of conjugate with protein, lipid and the like. Polysaccharide substances are widely present in plants, and researches show that nearly hundred kinds of polysaccharides are separated from plants at present, have no cytotoxicity and can be applied to organisms.
The carrageenan polysaccharide extract has the functions of resisting tumor, ulcer, virus, bacteria, blood sugar and blood fat, has antioxidant potential, and can be applied to the fields of food and medicine. The extraction method of carrageenan commonly used in the prior art comprises a dilute alkali solution extraction method, a dilute acid solution extraction method, a hot water extraction method and the like. However, due to the different conditions of reagents, temperature, etc., the chemical components of the carrageenan extracts obtained by different extraction methods are different, so that the efficacy of the carrageenan extracts is also different. For example, CN111743924A discloses a marine plant extract for treating diabetes, which is prepared from total polyphenol and/or polysaccharide of carrageenin and has the effect of inhibiting the activity of diabetes target alpha-glucosidase. CN100513427C discloses the manufacture of carrageenan and carrageenan products which produce carrageenan solutions that thicken, suspend, stabilize microparticles, colloidal dispersions and water/oil emulsions while also assisting in giving the food product a suitable mouthfeel.
At present, the prior art does not disclose a method for preparing a carrageenan extract which is simple to operate and has a remarkable antioxidant effect.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a carrageenan extract, a preparation method and an application thereof, wherein the preparation method is simple to operate, and the prepared carrageenan extract has remarkable antioxidant effect.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, the present invention provides a process for the preparation of a carrageenan extract, said process comprising the steps of:
(1) drying, pulverizing and sieving carrageen to obtain carrageen algae powder;
(2) carrying out reflux degreasing and drying on the carrageenin algae powder obtained in the step (1) to obtain degreased algae powder;
(3) carrying out water bath treatment on the degreased algae powder obtained in the step (2) to obtain a first mixed solution;
(4) centrifuging the first mixed solution obtained in the step (3), concentrating the supernatant, and adding a solvent to remove algin to obtain a second mixed solution;
(5) centrifuging the second mixed solution obtained in the step (4), concentrating the supernatant, adding a solvent, and precipitating to obtain a third mixed solution;
(6) centrifuging the third mixed solution obtained in the step (5), and collecting precipitates to obtain the carrageenan extract.
The preparation method of the carrageenan extract provided by the invention is simple to operate, and the prepared carrageenan extract has a remarkable antioxidant effect and can be applied to the field of food additives or pharmaceutical additives.
Preferably, the drying in step (1) is performed in a freeze drying manner, the drying time is 20-30h, for example, 20 h, 21h, 22 h, 23 h, 24 h, 25 h, 26 h, 27 h, 28 h, 29 h, 30h, and the like, and other specific values in the numerical range can be selected at will, which is not described herein again.
Preferably, the mesh number of the screen in the screening in the step (1) is 40-80 meshes, for example, 40 meshes, 45 meshes, 50 meshes, 55 meshes, 60 meshes, 65 meshes, 70 meshes, 75 meshes, 80 meshes, and the like, and other specific point values in the numerical range can be selected at will, and are not described in detail herein.
Preferably, the solvent used in the refluxing in step (2) comprises any one or a combination of at least two of ethanol, acetone, ethyl acetate, diethyl ether, methanol or petroleum ether.
The combination of at least two of the above-mentioned compounds, such as the combination of ethanol and acetone, the combination of acetone and ethyl acetate, the combination of ethyl acetate and ethyl ether, the combination of methanol and petroleum ether, etc., can be selected in any combination manner, and will not be described herein any more, and the combination of ethanol, ethyl ether and ethyl acetate is preferred.
The use of a combination of ethanol, diethyl ether and ethyl acetate as solvent for the reflux defatting of carrageenan powder compared to other solvents has surprisingly the effect of further improving the antioxidant capacity of the carrageenan extract produced.
Preferably, the refluxing time in step (2) is 1-4 times, such as 1 time, 2 times, 3 times, 4 times, and each refluxing time is 3-12h, such as 3 h, 3.5 h, 4 h, 4.5 h, 5 h, 5.5 h, 6h, 6.5 h, 7h, 7.5 h, 8 h, 8.5 h, 9 h, 9.5 h, 10 h, 10.5 h, 11 h, 11.5 h, 12h, etc., and other specific values in the value range can be selected, which is not repeated herein.
Preferably, the drying in step (2) is performed at 30 ℃ to 60 ℃, for example, 30 ℃, 32 ℃, 35 ℃, 37 ℃, 40 ℃, 42 ℃, 45 ℃, 47 ℃, 50 ℃, 52 ℃, 55 ℃, 57 ℃, 60 ℃ and the like, and other specific values in the numerical range can be selected, and are not described in detail herein.
Preferably, the water bath treatment in the step (3) is performed under the assistance of ultrasonic waves, the ultrasonic time is 40-80min, such as 40 min, 45 min, 50 min, 55 min, 60 min, 65 min, 70 min, 75 min, 80min, and the like, and other specific point values within the numerical range can be selected at will, and are not repeated herein.
Preferably, the liquid-to-material ratio in the water bath treatment in step (3) is 30-70mL/g, for example, 30 mL/g, 35 mL/g, 40 mL/g, 45 mL/g, 50 mL/g, 55 mL/g, 60 mL/g, 65 mL/g, 70mL/g, and the like, and other specific values in the numerical range can be selected, which is not described herein again.
Preferably, the temperature of the water bath treatment in the step (3) is 70-100 ℃, and the time of the water bath treatment is 3-7 h.
The specific value of 70-100 deg.C can be selected from 70 deg.C, 72 deg.C, 75 deg.C, 77 deg.C, 80 deg.C, 82 deg.C, 85 deg.C, 87 deg.C, 90 deg.C, 92 deg.C, 95 deg.C, 97 deg.C, 100 deg.C, etc.
The specific value of 3-7h can be selected from 3 h, 3.5 h, 4 h, 4.5 h, 5 h, 5.5 h, 6h, 6.5 h, 7h, etc.
Other specific point values within the above numerical ranges can be selected at will, and are not described in detail herein.
Preferably, the centrifugation in the step (4) is carried out at 15-40 ℃, the speed of the centrifugation is 3000-5000r/min, and the time of the centrifugation is 10-30 min.
The specific value of 15-40 deg.C can be selected from 15 deg.C, 17 deg.C, 20 deg.C, 22 deg.C, 25 deg.C, 27 deg.C, 30 deg.C, 32 deg.C, 35 deg.C, 37 deg.C, 40 deg.C, etc.
The specific value of 3000-5000r/min can be selected from 3000 r/min, 3200 r/min, 3500 r/min, 3700 r/min, 4000 r/min, 4200 r/min, 4500 r/min, 4700 r/min, 5000r/min, etc.
The specific value of 10-30min can be selected from 10 min, 12 min, 15 min, 17 min, 20 min, 22 min, 25 min, 27 min, 30min, etc.
Other specific point values within the above numerical ranges can be selected at will, and are not described in detail herein.
Preferably, the final volume of the concentration in step (4) is 1/6-1/4 times of the volume of the supernatant, such as 1/6 times, 1/5.8 times, 1/5.5 times, 1/5.2 times, 1/5 times, 1/4.8 times, 1/4.5 times, 1/4.2 times, 1/4 times, and the like, and other specific points in the value range can be selected, and are not repeated herein.
Preferably, the solvent in step (4) is 20% to 50% ethanol, and the 20% to 50% is a volume fraction of ethanol, such as 20%, 22%, 25%, 27%, 30%, 32%, 35%, 37%, 40%, 42%, 45%, 47%, 50%, and the like, and other specific points in the numerical range can be selected, and are not described herein, and preferably 25% to 35% ethanol is preferred.
Compared with ethanol with other concentrations, the removal effect is the best when 25% -35% ethanol is used as a solvent to remove algin, and further the antioxidant capacity of the prepared carrageenan extract can be further improved.
Preferably, the solvent removal of the algin in the step (4) is carried out at 15-25 ℃ for 0.5-2 h.
The specific value of the temperature of 15-25 deg.C can be selected from 15 deg.C, 16 deg.C, 17 deg.C, 18 deg.C, 19 deg.C, 20 deg.C, 21 deg.C, 22 deg.C, 23 deg.C, 24 deg.C, 25 deg.C, etc.
Specific values of 0.5-2h can be selected from 0.5 h, 0.6 h, 0.7 h, 0.8 h, 0.9 h, 1h, 1.1 h, 1.2 h, 1.3 h, 1.4 h, 1.5 h, 1.6 h, 1.7 h, 1.8 h, 1.9 h, 2h and the like.
Other specific point values within the above numerical ranges can be selected at will, and are not described in detail herein.
Preferably, the centrifugation in step (5) is performed at 15-40 ℃, the speed of the centrifugation is 3000-5000r/min, and the time of the centrifugation is 10-30 min.
The specific value of 15-40 deg.C can be selected from 15 deg.C, 17 deg.C, 20 deg.C, 22 deg.C, 25 deg.C, 27 deg.C, 30 deg.C, 32 deg.C, 35 deg.C, 37 deg.C, 40 deg.C, etc.
The specific value of 3000-5000r/min can be selected from 3000 r/min, 3200 r/min, 3500 r/min, 3700 r/min, 4000 r/min, 4200 r/min, 4500 r/min, 4700 r/min, 5000r/min, etc.
The specific value of 10-30min can be selected from 10 min, 12 min, 15 min, 17 min, 20 min, 22 min, 25 min, 27 min, 30min, etc.
Other specific point values within the above numerical ranges can be selected at will, and are not described in detail herein.
Preferably, the final volume of the concentration in step (5) is 1/6-1/4 times of the volume of the supernatant, such as 1/6 times, 1/5.8 times, 1/5.5 times, 1/5.2 times, 1/5 times, 1/4.8 times, 1/4.5 times, 1/4.2 times, 1/4 times, and the like, and other specific points in the value range can be selected, and are not repeated herein.
Preferably, the solvent in step (5) is 80% -95% ethanol, and the 80% -95% is a volume fraction of ethanol, such as 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, and the like, and other specific points in the numerical range can be selected, which is not described herein again.
Preferably, the volume of the solvent in the step (5) is 3-5 times of the volume of the supernatant after concentration, and the precipitation in the step (5) is carried out at 0-8 ℃ for 12-16 h.
The specific numerical range of 3 to 5 times may be 3 times, 3.2 times, 3.4 times, 3.6 times, 3.8 times, 4 times, 4.2 times, 4.4 times, 4.6 times, 4.8 times, 5 times, or the like.
The specific value range of 0-8 deg.C can be selected from 0 deg.C, 1 deg.C, 2 deg.C, 3 deg.C, 4 deg.C, 5 deg.C, 6 deg.C, 7 deg.C, 8 deg.C, etc.
The specific numerical range of 12-16h can be selected from 12h, 12.5 h, 13 h, 13.5 h, 14 h, 14.5 h, 15 h, 15.5 h, 16h and the like.
Other specific point values within the above numerical ranges can be selected at will, and are not described in detail herein.
Preferably, the centrifugation in the step (6) is carried out at 15-40 ℃, and the speed of the centrifugation is 3000-; the centrifugation time is 10-30 min.
The specific value of 15-40 deg.C can be selected from 15 deg.C, 17 deg.C, 20 deg.C, 22 deg.C, 25 deg.C, 27 deg.C, 30 deg.C, 32 deg.C, 35 deg.C, 37 deg.C, 40 deg.C, etc.
The specific value of 3000-5000r/min can be selected from 3000 r/min, 3200 r/min, 3500 r/min, 3700 r/min, 4000 r/min, 4200 r/min, 4500 r/min, 4700 r/min, 5000r/min, etc.
The specific value of 10-30min can be selected from 10 min, 12 min, 15 min, 17 min, 20 min, 22 min, 25 min, 27 min, 30min, etc.
Other specific point values within the above numerical ranges can be selected at will, and are not described in detail herein.
Preferably, the step (6) of collecting the precipitate further comprises washing the precipitate, wherein the washing reagent is absolute ethyl alcohol.
Preferably, the number of washes is 1-3, such as 1, 2, 3.
Preferably, the washing further includes drying the precipitate, the drying is performed in a freeze-drying manner, the drying time is 0.5-1h, for example, 0.5 h, 0.6 h, 0.7 h, 0.8 h, 0.9 h, 1h, and the like, and other specific values in the value range can be selected, which is not described herein again.
In a second aspect, the present invention provides a carrageenan extract produced by the method of producing a carrageenan extract according to the first aspect.
In a third aspect, the present invention provides the use of a carrageenan extract as described in the second aspect for the preparation of a food additive or a pharmaceutical additive.
Compared with the prior art, the invention has the following beneficial effects:
the preparation method of the carrageenan extract provided by the invention is simple to operate, and the prepared carrageenan extract has a remarkable antioxidant effect and can be applied to the field of food additives or pharmaceutical additives.
Detailed Description
To further illustrate the technical means and effects of the present invention, the following further describes the technical solution of the present invention with reference to the preferred embodiments of the present invention, but the present invention is not limited to the scope of the embodiments.
The following examples, comparative examples refer to carrageenans produced in Qinghai plateau from Roswent agricultural specialties.
Example 1
This example provides a method for preparing a carrageenan extract, comprising the steps of:
(1) freeze drying carrageen for 24 hr, pulverizing, and sieving with 60 mesh sieve to obtain carrageen algae powder;
(2) refluxing the carrageenin powder obtained in the step (1) for 3 times, wherein the refluxing time is 4 h each time, and drying at 40 ℃ to obtain degreased algae powder;
the solvent used in the reflux is 95% ethanol, the 95% ethanol is the volume fraction of the ethanol, and the liquid-material ratio of the solvent to the carrageenan powder is 15 mL/g;
(3) carrying out water bath treatment on the degreased algae powder obtained in the step (2) for 6 hours at 100 ℃ to obtain a first mixed solution;
the water bath treatment is carried out under the assistance of ultrasonic waves, the ultrasonic power is 300 w, the ultrasonic time is 50 min, and the liquid-material ratio of the water bath treatment is 50 mL/g;
(4) centrifuging the first mixed solution obtained in the step (3) at 25 ℃ at the rotating speed of 4000 r/min for 20 min, concentrating the supernatant to 1/5 volume, and treating the supernatant with 30% ethanol at 20 ℃ for 1h to obtain a second mixed solution;
(5) centrifuging the second mixed solution obtained in the step (4) at the rotating speed of 4000 r/min for 20 min at the temperature of 20 ℃, concentrating the supernatant to 1/5 volume, and treating the supernatant with 95% ethanol of 4 times of the volume of the concentrated supernatant for 12h at the temperature of 4 ℃ to obtain a third mixed solution;
(6) centrifuging the third mixed solution obtained in step (5) at 25 deg.C at 4000 r/min for 20 min, washing the precipitate with anhydrous ethanol for 3 times, and freeze drying for 0.5 hr to obtain carrageenan extract.
Example 2
This example provides a method for preparing a carrageenan extract, comprising the steps of:
(1) freeze drying carrageen for 28 hr, pulverizing, and sieving with 70 mesh sieve to obtain carrageen algae powder;
(2) refluxing the carrageenin powder obtained in the step (1) for 2 times, wherein the refluxing time is 6h each time, and drying at 55 ℃ to obtain degreased algae powder;
the solvent used in the reflux is absolute methanol, and the liquid-material ratio of the absolute methanol to the carrageenin powder is 15 mL/g;
(3) carrying out water bath treatment on the degreased algae powder obtained in the step (2) for 5 hours at 90 ℃ to obtain a first mixed solution;
the water bath treatment is carried out under the assistance of ultrasonic waves, the ultrasonic power is 300 w, the ultrasonic time is 60 min, and the liquid-material ratio of the water bath treatment is 60 mL/g;
(4) centrifuging the first mixed solution obtained in the step (3) at the rotating speed of 4500 r/min for 15 min at the temperature of 15 ℃, concentrating the supernatant to 1/6 volume, and treating with 20% ethanol at the temperature of 20 ℃ for 2h to obtain a second mixed solution;
(5) centrifuging the second mixed solution obtained in the step (4) at 35 ℃ at a rotating speed of 3000 r/min for 30min, concentrating the supernatant to 1/6 volume, and treating the supernatant with 90% ethanol in volume which is 3 times of the concentrated supernatant at 0 ℃ for 14 h to obtain a third mixed solution;
(6) centrifuging the third mixed solution obtained in step (5) at 15 deg.C at 5000r/min for 10 min, washing the precipitate with anhydrous ethanol for 2 times, and freeze drying for 0.5 hr to obtain carrageenan extract.
Example 3
This example provides a method for preparing a carrageenan extract, comprising the steps of:
(1) freeze drying carrageen for 21 hr, pulverizing, and sieving with 50 mesh sieve to obtain carrageen algae powder;
(2) refluxing the carrageenin powder obtained in the step (1) for 1 time, wherein the refluxing time is 12h, and drying at 35 ℃ to obtain degreased algae powder;
the solvent used in the reflux is anhydrous acetone, and the liquid-material ratio of the anhydrous acetone to the carrageenan powder is 15 mL/g;
(3) carrying out water bath treatment on the degreased algae powder obtained in the step (2) for 7 hours at the temperature of 80 ℃ to obtain a first mixed solution;
the water bath treatment is carried out under the assistance of ultrasonic waves, the ultrasonic power is 300 w, the ultrasonic time is 40 min, and the liquid-material ratio of the water bath treatment is 70 mL/g;
(4) centrifuging the first mixed solution obtained in the step (3) at the temperature of 30 ℃ at the rotating speed of 3500 r/min for 25 min, concentrating the supernatant to 1/4 volume, and treating with 50% ethanol at the temperature of 20 ℃ for 0.5 h to obtain a second mixed solution;
(5) centrifuging the second mixed solution obtained in the step (4) at the rotating speed of 4500 r/min for 15 min at 15 ℃, concentrating the supernatant to 1/4 volume, and treating the supernatant with 80% ethanol of 5 times of the volume of the concentrated supernatant for 16h at 8 ℃ to obtain a third mixed solution;
(6) centrifuging the third mixed solution obtained in step (5) at 35 deg.C at 3000 r/min for 30min, washing the precipitate with anhydrous ethanol for 2 times, and freeze drying for 0.5 hr to obtain carrageenan extract.
Example 4
This example provides a process for the preparation of a carrageenan extract which differs from example 1 only in that the solvent "95% ethanol" used in the reflux in step (2) is replaced by the same amount of "dehydrated ether" added, otherwise the conditions are as in example 1.
Example 5
This example provides a process for the preparation of a carrageenan extract, which differs from example 1 only in that the solvent "95% ethanol" used in the reflux in step (2) is replaced by "anhydrous ethyl acetate" in the same amount, and otherwise the conditions are as in example 1.
Example 6
This example provides a process for the preparation of a carrageenan extract which differs from example 1 only in that the solvent "95% ethanol" used in the reflux in step (2) is replaced by the same amount of "a combination of 95% ethanol and anhydrous ethyl acetate" wherein the volume ratio of 95% ethanol to anhydrous ethyl acetate is 1:1, and otherwise the conditions refer to example 1.
Example 7
This example provides a process for the preparation of a carrageenan extract which differs from example 1 only in that the solvent "95% ethanol" used in the reflux in step (2) is replaced by the same amount of "a combination of 95% ethanol and anhydrous diethyl ether" wherein the volume ratio of 95% ethanol to anhydrous diethyl ether is 1:1, and otherwise the conditions are as in example 1.
Example 8
This example provides a process for the preparation of a carrageenan extract which differs from example 1 only in that the solvent "95% ethanol" used in the reflux in step (2) is replaced by the same amount of "a combination of anhydrous ethyl acetate and anhydrous diethyl ether" wherein the volume ratio of anhydrous ethyl acetate to anhydrous diethyl ether is 1:1, and otherwise the conditions are as in example 1.
Example 9
This example provides a process for the preparation of a carrageenan extract which differs from example 1 only in that the solvent "95% ethanol" used in the refluxing in step (2) is replaced by the same amount of "a combination of 95% ethanol, anhydrous ethyl acetate and anhydrous diethyl ether" wherein the volume ratio of 95% ethanol, anhydrous ethyl acetate and anhydrous diethyl ether is 1:1:1, and the other conditions refer to example 1.
Example 10
This example provides a method for preparing a carrageenan extract, which is different from example 1 only in that the solvent "30% ethanol" in step (4) is replaced by the same amount of "20% ethanol", and other conditions refer to example 1.
Example 11
This example provides a method for preparing a carrageenan extract, which is different from example 1 only in that the solvent "30% ethanol" in step (4) is replaced with the same amount of "50% ethanol", and the other conditions refer to example 1.
Comparative example 1
This comparative example provides a method for preparing a carrageenan extract, which is different from example 1 only in that the reflux defatting operation in step (2) is omitted, and the other conditions refer to example 1.
Test example 1
The extraction yield of the carrageenan extracts prepared in examples 1-11 and comparative example 1 was evaluated.
The total sugar content is determined by phenol-sulfuric acid method, polysaccharide is hydrolyzed into monosaccharide under the action of sulfuric acid, and is rapidly dehydrated to generate uronic acid derivative, which reacts with phenol to generate orange yellow substance with characteristic absorption at 490 nm, and the content is quantified compared with standard series. The carrageenan content was tested with glucose as standard.
The specific operation steps are as follows;
(1) dissolving 1 mg of the prepared carrageenan extract in 1 mL of distilled water to prepare a solution to be detected;
(2) transferring all the solution to be detected obtained in the step (1) to a 20 mL glass test tube with a plug, supplementing the solution to 1 mL by using distilled water, and adding 1 mL of phenol (5%);
(3) adding 5 mL concentrated sulfuric acid (vertical to the liquid surface, without contacting the wall of the test tube, to mix with the reaction solution), and standing for 10 min;
(4) the reaction solution was thoroughly mixed using a vortex shaker, and then the tube was placed in a water bath at 30 ℃ for 20 min, and the absorbance was measured at 490 nm.
The polysaccharide content in the sample was calculated according to the following formula:
Figure DEST_PATH_IMAGE002
wherein: m is1-the polysaccharide content corresponds to the sugar content (ug) on the standard curve;
V1-sample volumetric volume (mL);
m2-sample mass (g);
V2- -volume of sample removed for colorimetric determination (mL);
0.9- - -conversion factor of glucose to dextran.
The results of comparing the extraction yields of the carrageenan extracts prepared in examples 1-11 and comparative examples 1 and 2 are shown in Table 1.
TABLE 1
Group of Extraction rate
Example 1 3.53%
Example 2 3.19%
Examples3 3.24%
Example 4 2.89%
Example 5 2.98%
Example 6 3.63%
Example 7 3.66%
Example 8 3.65%
Example 9 3.83%
Example 10 3.05%
Example 11 3.01%
Comparative example 1 1.03%
The results show that: the carrageenans extracts prepared in examples 1-11 were obtained with higher extraction yields, with the carrageenans extracts prepared in examples 6-8 being slightly higher than in example 1, the carrageenans extract prepared in example 9 being the highest extraction yield, the carrageenans extracts prepared in examples 10 and 11 being slightly lower than in example 1 and the carrageenans extract prepared in comparative example 1 being lower extraction yields.
Test example 2
The antioxidant effect of the carrageenan extracts prepared in examples 1-11 and comparative example 1 was evaluated using an in vitro test. The test method is as follows:
dissolving carrageenan extract in anhydrous ethanol to prepare a series of sample solutions to be tested, wherein the sample solutions are 1 mg/mL, 2 mg/mL, 4 mg/mL, 8 mg/mL, 16 mg/mL and 24 mg/mL.
(1) DPPH radical scavenging ability test:
preparing 0.2 mmol/L DPPH solution (the solvent is absolute ethyl alcohol);
sample measurement A1: mixing 100 μ L sample solution to be detected and 100 μ L DPPH solution in 96-well plate, reacting in dark for 30min, measuring the light absorption value at 517 nm, and recording as A1；
Sample blank A2: mixing 100 μ L sample solution to be tested with 100 μ L anhydrous ethanol in 96-well plate, reacting in dark for 30min, measuring light absorption value at wavelength of 517 nm, and recording as A2；
Reagent blank A0: mixing 100 μ L DPPH solution and 100 μ L anhydrous ethanol in 96-well plate, reacting in dark for 30min, measuring the absorbance at 517 nm, and recording as A0；
Figure DEST_PATH_IMAGE004
(2) ABTS free radical scavenging ability test:
preparing 5 mmol/L ABTS stock solution, diluting with anhydrous ethanol to make the absorbance value of the diluted solution at 734 nm be 0.7 + -0.02 (at 30 deg.C), and obtaining ABTS working solution.
Sample measurement A1: absorbing 190 μ L ABTS working solution, adding 10 μ L sample solution to be tested, oscillating for 10 s, standing at 30 deg.C for 6 min, measuring absorbance at 734 nm, and recording as A1；
Sample blank A2: absorbing 190 mu L of absolute ethyl alcohol, and adding 10 mu L of absolute ethyl alcohol to be treatedMeasuring the sample solution, oscillating for 10 s, standing at 30 deg.C for 6 min, measuring the light absorption value at 734 nm, and recording as A2；
Reagent blank A0: absorbing 190 μ L ABTS working solution, adding 10 μ L anhydrous ethanol, oscillating for 10 s, standing at 30 deg.C for 6 min, measuring absorbance at 734 nm, and recording as A0；
Figure DEST_PATH_IMAGE005
(3) Hydroxyl radical scavenging ability test:
sample measurement A1: add 60. mu.L of FeSO to 96-well plates4Solution, 60 μ L salicylic acid solution, and finally 60 μ L H2O2Starting the reaction of the solution, uniformly mixing, and incubating for 15 min at 37 ℃; adding 20 μ L of sample solution to be tested, incubating at 37 deg.C for 15 min, measuring absorbance at 510 nm, and recording as A1；
Sample blank A2: add 60. mu.L of FeSO to 96-well plates4The solution is 60 mu L of salicylic acid solution, and finally 60 mu L of distilled water is added, and after uniform mixing, incubation is carried out for 15 min at 37 ℃; adding 20 μ L of sample solution to be tested, incubating at 37 deg.C for 15 min, measuring absorbance at 510 nm, and recording as A2；
Reagent blank A0: add 60. mu.L of FeSO to 96-well plates4Solution, 60 μ L salicylic acid solution, and finally 60 μ L H2O2Starting the reaction of the solution, uniformly mixing, and incubating for 15 min at 37 ℃; adding distilled water 20 μ L, incubating at 37 deg.C for 15 min, measuring absorbance at 510 nm, and recording as A0；
Figure DEST_PATH_IMAGE007
And (3) drawing a change curve by taking the concentration of the sample solution to be detected as an abscissa and the free radical scavenging rate as an ordinate so as to calculate the IC50 value of the carrageenan extract for scavenging free radicals.
The test results are shown in Table 2.
TABLE 2
Group of IC50 (mg/mL) for scavenging DPPH free radicals IC50 (mg/mL) for scavenging ABTS free radicals IC50 (mg/mL) for scavenging hydroxy radicals
Example 1 8.16 17.02 8.81
Example 2 8.32 17.56 9.04
Example 3 8.55 17.87 8.93
Example 4 8.93 17.85 9.78
Example 5 9.03 17.95 9.96
Example 6 7.65 16.57 8.23
Example 7 7.57 16.43 8.16
Example 8 7.63 16.68 8.34
Example 9 7.02 16.07 7.75
Example 10 8.56 17.34 9.23
Example 11 8.64 17.54 9.41
Comparative example 1 11.23 20.02 11.87
The results show that: the carrageenan extracts prepared in examples 1-11 have a strong scavenging ability against DPPH free radicals, ABTS free radicals and hydroxyl free radicals, i.e. a significant antioxidant effect. Among them, the carrageenan extracts obtained in examples 6 to 8 had slightly better antioxidant effect than that of example 1, the carrageenan extract obtained in example 9 had the best antioxidant effect, the carrageenan extracts obtained in examples 10 and 11 had slightly worse antioxidant effect than that of example 1, and the carrageenan extract obtained in comparative example 1 had poor antioxidant effect.
Test example 3
The antioxidant effect of the carrageenan extracts prepared in examples 1 to 11 and comparative example 1 was evaluated using an in vivo experiment based on a caenorhabditis elegans (hereinafter, referred to as nematode) model. The test method is as follows:
(1) preparing a nematode culture medium and a reagent:
(1M) Potassium phosphate buffer
KH2PO4 108.39 g
K2HPO4 35.69 g
Adding water to 1L, and adjusting pH to 6.0.
② M9 buffer solution
Na2HPO4 6 g
KH2PO4 3 g
NaCl 5 g
MgSO4·7H2O 0.25 g
Adding water to 1L, and sterilizing at 121 deg.C for 15 min.
③ LB liquid culture medium
Tryptone 10 g/L
Sodium chloride 10 g/L
Yeast powder 5 g/L
Preparing distilled water, adjusting pH to 7.0 with 1M sodium hydroxide solution, and sterilizing at 121 deg.C for 15 min.
(iv) Nematode Growth solid Medium (NGM) 1L
NaCl 3 g
Agar 17 g
TryPtone (TryPtone) 2.5 g
Streptomyces (Streptomycin) 0.2 g
975 mL of distilled water
Shaking, sterilizing at 121 deg.C for 30min, keeping the temperature at 80 deg.C for 15 min, and adding the following sterilized solutions (wherein cholesterol is sterilized by filtration, and the rest solutions are sterilized at high temperature).
1 M CaCl2 l mL
1 M MgSO4 l mL
5 mg/mL Cholesterol (solvent ethanol) l mL
25 mL of 1M potassium phosphate buffer
Lysate
0.1g NaOH and 1.3 mL NaClO were dissolved in 4 mL distilled water and mixed well (ready for use).
(2) Basic nematode manipulation method
(ii) cultivation of Escherichia coli OP50
Taking OP50 strain to streak on LB plate, picking single colony in 10 mL LB liquid culture medium, culturing at 37 deg.C and 200 rpm under shaking for 12h to OD600Equal to 0.4, for inoculation of NGM to feed normal group nematodes.
② E.coliApplication of OP50
An appropriate amount of inoculum (typically 60 mm diameter plate plus 100. mu.L) was added to each NGM plate and the inoculum was spread evenly on the NGM plate using a sterile spreader or the bottom of a glass test tube, taking care that the inoculum edge should be about 0.5 cm from the plate edge. The bacterial-coated NGM plates were kept overnight at room temperature (21-25 deg.C) in a cold room or 4 deg.C freezer until use.
③ synchronization of nematodes
Washing young adults into an aseptic EP tube by using M9 buffer solution, adding a proper amount of lysate to crack nematodes, placing the nematode on a low-speed centrifuge at 3000 rpm for 1 min, discarding the supernatant, washing the nematodes for 2 times by using M9, centrifuging the nematode, discarding the supernatant, sucking the nematodes at the bottom of the EP tube by using a pipette gun to drip into an aseptic area of the NGM, basically developing fertilized eggs in the cracked nematodes into L4 larvae after about 48 hours, and completing synchronization for the experiment.
Cracking of nematode
The young adults on the NGM plate are repeatedly washed twice by 1 mL of M9 buffer solution and then transferred to a sterile 2 mL centrifuge tube, 1 mL of prepared lysate is added, after full oscillation for 3-5 min, the mixture is centrifuged for 1 min at the rotating speed of 3000 rpm, and the supernatant is discarded. The nematodes were washed with 1 mL of M9 buffer, centrifuged 2 times under the same conditions, the supernatant discarded, and 0.3-0.4 mL of egg-containing buffer remained. Then gently blow and beat the mixed eggs by using a pipette gun, and suck about 100 mu L of buffer solution containing the eggs to a sterile area close to OP50 on an NGM plate. After about 48 hours, the nematode oosperm basically develops into L4 larvae, and synchronization is completed. The larvae in stage L4 were picked and transferred to drug plates for each index measurement.
(3) Preparation of carrageenan-containing NGM:
dissolving 1 mg carrageenin in 1 mL sterile water to obtain mother solution, and collecting 98 μ LE.coliOP50 bacterial liquid and 2 mul mother liquid are mixed evenly. A pipette gun was used to aspirate 150. mu.L of mixed inoculum and gently pipette the mixed inoculum into the center of each NGM plate. Blank group extraction 100-E.coliOP50 bacterial liquid is uniformly coated. And (4) drying the bacterial-coated NGM plate in the dark, sealing the membrane, and storing at 4 ℃ for later use.
(4) And (3) accumulation test of active oxygen in the nematode:
the placebo and the test group after 96 h carrageenan treatment were transferred to NGM plates to remove e. After 3 transfers, the nematodes were transferred to 96-well fluorescent plates containing 50. mu. L M9 buffer, 80 per well, in triplicate. At the same time, 50. mu.L of 100. mu.M H in M9 buffer was added2DCF-DA solution with 50. mu.L of H without nematodes2DCF-DA solution and 50. mu. L M9 buffer served as blanks.Immediately, the 96-well plate is placed into an enzyme-labeling instrument to measure the fluorescence intensity, the reaction temperature is 25 ℃, the emission wavelength is 528 nm, and the excitation wavelength is 485 nm. The assay was performed every 20 min for 6h with shaking before each reading. Because the accumulation of ROS in the nematode is in direct proportion to the generation amount of the fluorescent substance DCF, the fluorescence intensity of each group can directly reflect the accumulation amount of ROS in the nematode.
(5) CAT activity test in the nematode body:
respectively selecting 200 nematodes from a blank group of nematodes and a test group of nematodes treated by carrageenan for 96 h, placing the nematodes into sterile water for washing, centrifuging at a low speed to remove supernatant, repeatedly carrying out three times, fixing the volume to 0.5 mL, and carrying out low-temperature grinding for 1 min in a grinding instrument. Grinding, adding 1% chap solution to a constant volume of 1 mL, centrifuging at low temperature (4 ℃, 12000 r/min, 15 min), collecting supernatant, and standing at 4 ℃ for testing. The determination of the enzyme activity is carried out according to the specification requirements of BCA and CAT test boxes of Nanjing institute of Biotechnology.
The test results are shown in Table 3.
TABLE 3
Group of Accumulated amount of ROS CAT Activity (U/mgprot)
Blank group (nematodes not treated with carrageenan extract) 1 ± 0.12 22.71 ± 0.96
Example 1 0.78 ± 0.31 29.76 ± 1.12
Example 2 0.80 ± 0.28 29.98 ± 1.83
Example 3 0.79 ± 0.07 29.85 ± 0.76
Example 4 0.85 ± 0.24 27.47 ± 2.17
Example 5 0.84 ± 0.19 27.56 ± 3.01
Example 6 0.72 ± 0.12 31.78 ± 0.86
Example 7 0.73 ± 0.17 31.17 ± 1.28
Example 8 0.72 ± 0.28 31.34 ± 2.02
Example 9 0.65 ± 0.07 33.72 ± 1.46
Example 10 0.82 ± 0.22 27.81 ± 2.27
Example 11 0.81 ± 0.14 28.02 ± 2.63
Comparative example 1 0.93 ± 0.09 24.12 ± 1.26
The results show that: the carrageenans extracts prepared in examples 1-11 all significantly reduced the accumulation of active oxygen in the nematode while significantly enhanced CAT activity in the nematode, i.e., had significant antioxidant effects. Among them, the carrageenan extracts obtained in examples 6 to 8 had slightly better antioxidant effect than that of example 1, the carrageenan extract obtained in example 9 had the best antioxidant effect, the carrageenan extracts obtained in examples 10 and 11 had slightly worse antioxidant effect than that of example 1, and the carrageenan extract obtained in comparative example 1 had poor antioxidant effect.
In conclusion, the preparation method of the carrageenan extract provided by the invention is simple to operate, and the prepared carrageenan extract has a remarkable antioxidant effect and can be applied to the field of food additives or pharmaceutical additives.
The applicant states that the present invention is illustrated by the above examples of the carrageenan extract and the method of preparation and use of the present invention, but the present invention is not limited to the above examples, i.e. it is not meant that the present invention must be practiced by relying on the above examples. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.
The preferred embodiments of the present invention have been described in detail, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention.
It should be noted that the various technical features described in the above embodiments can be combined in any suitable manner without contradiction, and the invention is not described in any way for the possible combinations in order to avoid unnecessary repetition.

Claims (3)

1. A method for preparing a carrageenan extract, wherein carrageen is produced from Qinghai plateau, the method for preparing the carrageenan extract comprises the following steps:
(1) freeze drying carrageen for 20-30 hr, pulverizing, and sieving with 40-80 mesh sieve to obtain carrageen algae powder;
(2) refluxing the carrageenin powder obtained in the step (1) for 1-4 times, wherein the refluxing time is 3-12h each time, and drying at 30-60 ℃ to obtain degreased algae powder;
the solvent used in the reflux is a combination of 95% ethanol, ethyl ether and ethyl acetate, wherein the volume ratio of the 95% ethanol to the anhydrous ethyl acetate to the anhydrous ethyl ether is 1:1: 1;
(3) treating the defatted algae powder obtained in the step (2) for 3-7h in water bath at 70-100 ℃ to obtain a first mixed solution;
the water bath treatment is carried out under the assistance of ultrasonic waves, the ultrasonic time is 40-80min, and the liquid-material ratio of the water bath treatment is 30-70 mL/g;
(4) centrifuging the first mixed solution obtained in the step (3) at the rotating speed of 3000-;
(5) centrifuging the second mixed solution obtained in the step (4) at the rotating speed of 3000-;
(6) centrifuging the third mixed solution obtained in the step (5) at the rotating speed of 3000-.
2. A carrageenan extract, characterized in that it has been prepared by the process for the preparation of a carrageenan extract according to claim 1.
3. Use of a carrageenan extract according to claim 1 for the preparation of a food additive or a pharmaceutical additive.
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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111454899A (en) * 2020-04-30 2020-07-28 青岛思拓新源细胞医学有限公司 Application of carrageenan in inhibiting mesenchymal stem cell lipogenesis transformation

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111454899A (en) * 2020-04-30 2020-07-28 青岛思拓新源细胞医学有限公司 Application of carrageenan in inhibiting mesenchymal stem cell lipogenesis transformation

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
"响应面法优化鹿角菜中岩藻多糖的超声波辅助提取工艺";宋海燕等;《食品与发酵工业》;20151103;第41卷(第12期);第212-218页 *
响应面法优化鹿角菜中岩藻多糖的提取工艺;宋海燕等;《海洋科学》;20160415;第40卷(第04期);第73-80页 *

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