CN110698695B - Preparation method of colloidal emulsion with stable cellulose nanofibers - Google Patents
Preparation method of colloidal emulsion with stable cellulose nanofibers Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/02—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques
- C08J3/09—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in organic liquids
- C08J3/091—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in organic liquids characterised by the chemical constitution of the organic liquid
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2301/00—Characterised by the use of cellulose, modified cellulose or cellulose derivatives
- C08J2301/02—Cellulose; Modified cellulose
Abstract
The invention discloses a preparation method of colloidal emulsion with stable cellulose nano-fiber, which comprises the following steps: taking mushroom stem powder as a raw material, and extracting cellulose; dissolving the cellulose in water, and preparing a cellulose nanofiber solution by utilizing high-pressure homogenization; adding vegetable oil into CNFs according to the weight ratio of 1 (1) - (3), and homogenizing for 2-20 times under 10-90 MPa to obtain the colloidal emulsion. The invention uses the fungal fruiting body cellulose nano-fiber for the first time to prepare the stable colloidal medium-internal phase emulsion, and the colloidal emulsion is obtained under the condition of 50% internal phase ratio. The raw materials used in the preparation method are food-grade materials, are rich in resources, have the advantages of environmental protection, reproducibility and low cost, and simultaneously have good biodegradability and biocompatibility, the preparation conditions of the colloidal medium-internal phase emulsion are mild, the operation is simple, the colloidal medium-internal phase emulsion can be stored for 30 days at 25 ℃ without phase separation, and the preparation method has great application value in the fields of food, medicine, cosmetics and the like.
Description
Technical Field
The invention belongs to the technical field of emulsion preparation, and particularly relates to a preparation method of colloidal medium-internal phase emulsion with stable cellulose nanofibers.
Background
The colloidal emulsion attracts the interest of a plurality of researchers due to the characteristics of both gel and emulsion, and is widely applied to the fields of food, medicine, cosmetics, 3D printing and the like. The colloidal network structure in the colloidal emulsion can improve the stability and antioxidant activity of the emulsion, thereby inhibiting the oxidation of unsaturated fatty acids in the emulsion, and the characteristic enables the colloidal emulsion to play an important role in a drug delivery system. Current research has demonstrated that particles such as whey protein, soy protein, porcine plasma protein, and zein can stabilize the oil-water interface in a colloidal emulsion to form a uniform colloidal emulsion. However, most of these particles are chemically modified protein particles, and are greatly affected by temperature and pH. Until now, the research on natural polymer-stabilized colloidal emulsions has been very limited.
The use of cellulose to produce colloidal emulsions has been less and less studied than protein. Cellulose is one of the most abundant natural resources on earth and is also a major constituent of terrestrial plants. A large number of research documents have demonstrated that cellulose can be extracted and modified in various ways and then used as an emulsifier in the food industry. Among them, the most widely used is nano-sized cellulose, such as Cellulose Nanofibers (CNFs). CNFs play an important role in natural nanoscale particles due to their high aspect ratio, high strength, and the ability to form networks in emulsions. In the existing literature reports, the natural CNFs having the function of stabilizing the oil-water interface are mostly obtained from wood or plants, such as mangosteen bark, cotton, Pinus radiate wood, etc., and these CNFs can stabilize 10% -35% of the internal phase in the emulsion to form the emulsion with high fluidity. However, these materials are derived from plant and wood cellulose, and are rich in a large amount of hard lignin, which affects the ability of CNFs to stabilize oil-water interface to some extent.
The mushroom is the fruiting body of the fungus of the Lentinus Edodes, is the second most edible fungus in the world, is one of the special products in China, and is called mountain delicacies among people. The mushroom has the function of decomposing lignin, the main edible part is the mushroom umbrella part, and the mushroom stem is usually discarded in the processing process due to the higher cellulose content and poor palatability, so that the mushroom becomes industrial waste and causes great resource waste. To our knowledge, the preparation of colloidal emulsions at the medium internal phase level has not been reported.
Disclosure of Invention
The invention aims to prepare a colloidal medium-internal phase emulsion by adopting mushroom stem CNFs as a disperse phase and adopting a high-pressure homogenization method. The invention utilizes the nanometer cellulose of the fungus fruiting body to stabilize the colloidal emulsion for the first time and obtains the food-grade colloidal emulsion under the condition of 50 percent internal phase ratio. The raw materials used in the preparation method are food-grade materials, are rich in resources, have the advantages of environmental protection, reproducibility and low cost, and simultaneously have good biodegradability and biocompatibility, the preparation condition of the colloidal emulsion is mild, the operation is simple, the colloidal emulsion has the characteristics of both emulsion and gel, and can be stably stored for 30 days at room temperature (25 ℃) without phase separation, thereby having great application value in the fields of food, medicine, cosmetics and the like.
In order to achieve the above object, the present invention provides a method for preparing a colloidal emulsion stabilized by cellulose nanofibers, comprising the steps of:
s1, extracting the lentinus edodes cellulose (native cellulose, NC): taking dried shiitake stipe powder, adding 300mg/mL sodium hydroxide solution with the weight 5-200 times of that of the dried shiitake stipe powder, boiling, keeping for 1-10 h, and filtering by a 20-180-mesh sieve to obtain a precipitate as crude cellulose; adding 9.97mol/L hydrogen peroxide solution which is 5-50 times of the weight of the crude cellulose into the crude cellulose, stirring the crude cellulose for 1 hour at the temperature of 30-70 ℃ and 1000rpm, stirring the crude cellulose for 1-24 hours at the temperature of 25 ℃ and 1000rpm, sieving the obtained mixture by a 20-180-mesh sieve, washing the obtained precipitate A by deionized water to the pH value of 7.0-7.9, and centrifuging the precipitate B to obtain the lentinus edodes cellulose; the mushroom cellulose is fibrous;
s2, preparing lentinus edodes stem Cellulose Nanofibers (CNFs): dissolving the lentinus edodes cellulose in water in the step S1 to prepare a solution with the cellulose concentration of 10-25 mg/mL, and homogenizing for 2-20 times under the high pressure of 10-90 MPa to obtain a cellulose-rice-fiber solution;
s3, preparing a colloidal emulsion: taking the cellulose nanofiber solution with the concentration of 10-25 mg/mL in the step S2, adding vegetable oil into the cellulose nanofiber solution, and homogenizing for 2-20 times under the high pressure of 10-90 MPa to obtain colloidal emulsion with stable Cellulose Nanofibers (CNFs); wherein the weight ratio of the cellulose nanofiber solution to the vegetable oil is 1 (1-3).
Preferably, the preparation method of the dried mushroom stem powder in step S1 includes: taking dried shiitake mushroom stems with moisture content of less than 13g/100g, and crushing the dried shiitake mushroom stems into 20-180 meshes of powder; the centrifugation is carried out at 1000-10000 rpm for 5-60 min; the dried mushroom stems conform to the national food safety standard of edible fungi and products GB7096-2014 thereof.
Preferably, the vegetable oil in step S3 is an edible oil such as soybean oil, salad oil, corn oil, sunflower seed oil or blend oil, and an aromatized vegetable oil obtained by extracting an edible oil as a base.
In a preferred embodiment, the preparation method of the colloidal emulsion stabilized by cellulose nanofibers comprises the following steps:
s1, extracting mushroom cellulose (NC): crushing dried lentinus edodes stems, filtering the crushed lentinus edodes stems through a 20-mesh sieve to obtain 10g of lentinus edodes stem powder, adding 2000g of 300mg/mL sodium hydroxide solution into the lentinus edodes stem powder, boiling the mixture for 4 hours, filtering the obtained mixture through a 120-mesh sieve to obtain crude cellulose, adding 10g of the obtained crude cellulose into 200g of 9.97mol/L hydrogen peroxide solution, stirring the mixture in a stirrer at 1000rpm for 1 hour at 70 ℃, stirring the mixture at 25 ℃ for 12 hours at 1000rpm, filtering the obtained mixture through the 120-mesh sieve to obtain a precipitate A, washing the precipitate A to be pH7.9 with deionized water, and centrifuging the precipitate B at 5000rpm for 30 minutes to obtain lentinus edodes cellulose (NC);
s2, preparing lentinus edodes stem Cellulose Nanofibers (CNFs): dissolving the lentinus edodes cellulose obtained in the step S1 in water to prepare a solution with the lentinus edodes cellulose concentration of 25mg/mL, and then homogenizing for 10 times under the high pressure of 80MPa to obtain a CNFs solution;
s3, preparing a colloidal emulsion: and (5) taking 50g of the CNFs solution obtained in the step S2, adding 50g of soybean oil into the CNFs solution, stirring to uniformly disperse the soybean oil, and homogenizing for 10 times under high pressure of 80MPa to obtain the colloidal emulsion with stable CNFs.
The beneficial effects of the invention are mainly embodied in the following four aspects:
firstly, the invention applies stable colloidal emulsion of fungus fruiting body CNFs for the first time, the lentinus edodes stem cellulose resource is rich, the lentinus edodes processing by-product is obtained, and the invention has the advantages of green environmental protection, regeneration and low cost, and simultaneously has good biodegradability and biocompatibility.
Secondly, the preparation condition of the colloidal emulsion is mild, the operation is simple, the colloidal emulsion has the double characteristics of emulsion and gel, the colloidal emulsion can be stably stored for 30 days at room temperature (25 ℃) without phase separation, and the colloidal emulsion has great application value in the fields of food, medicine, cosmetics and the like.
Thirdly, the invention realizes the preparation of the colloidal emulsion by using the fungal nano-cellulose at the level of the middle-internal phase with the internal phase mass ratio of 50%.
Fourthly, the CNFs extracted from the lentinus edodes stems by the method are dendritic, and can form a gel network in the emulsion, so that the formation of the gel emulsion is promoted, and the stability of the emulsion is further improved.
Drawings
FIG. 1 is a diagram showing the appearance of the colloidal emulsion obtained in example 1 of the present invention.
FIG. 2 is an appearance diagram of a colloidal emulsion obtained in example 2 of the present invention.
FIG. 3 is an appearance diagram of a colloidal emulsion obtained in example 3 of the present invention.
FIG. 4 is an appearance diagram of a colloidal emulsion obtained in example 4 of the present invention.
FIG. 5 is an appearance diagram of a colloidal emulsion obtained in comparative example 1 of the present invention.
FIG. 6 is an appearance of the colloidal emulsion obtained in comparative example 2 of the present invention.
FIG. 7 is an appearance of a colloidal emulsion obtained in comparative example 3 of the present invention.
FIG. 8 is a microscopic image of the colloidal emulsion obtained in example 1 of the present invention under a confocal laser scanning microscope.
FIG. 9 is a microscopic image of the colloidal emulsion obtained in example 2 of the present invention under a confocal laser scanning microscope.
FIG. 10 is a microscopic image of the colloidal emulsion obtained in example 3 of the present invention under a confocal laser scanning microscope.
FIG. 11 is a microscopic image of the colloidal emulsion obtained in example 4 of the present invention under a confocal laser scanning microscope.
FIG. 12 is a microscopic image of the colloidal emulsion obtained in comparative example 1 of the present invention under a confocal laser microscope.
FIG. 13 is a microscopic image of the colloidal emulsion obtained in comparative example 2 of the present invention under a confocal laser microscope.
FIG. 14 is a microscopic morphology image under a scanning electron microscope of the colloidal emulsion obtained in example 1 of the present invention.
FIG. 15 is a microscopic image of the colloidal emulsion obtained in example 2 of the present invention under a scanning electron microscope.
FIG. 16 is a microscopic morphology image under a scanning electron microscope of the colloidal emulsion obtained in example 3 of the present invention.
FIG. 17 is a microscopic morphology image under a scanning electron microscope of the colloidal emulsion obtained in example 4 of the present invention.
FIG. 18 is a microscopic morphology under a scanning electron microscope of the colloidal emulsion obtained in comparative example 1 of the present invention.
FIG. 19 is a microscopic morphology image under a scanning electron microscope of the colloidal emulsion obtained in comparative example 2 of the present invention.
FIG. 20 is a graph showing the thiobarbituric acid values of the colloidal emulsions obtained in examples 1 to 4 of the present invention and comparative examples 1 to 2 during 30-day storage, wherein the lower case letters a to e indicate significant differences (p < 0.05).
FIG. 21 shows the emulsification indexes of the colloidal emulsions obtained in examples 1 to 4 of the present invention and comparative examples 1 to 2.
Detailed Description
The present invention is further illustrated by the following specific examples, but the embodiments of the present invention are not limited thereto.
The invention provides a preparation method of a stable colloidal emulsion of mushroom stem CNFs, which comprises the following steps:
s1, extracting NC: crushing the lentinus edodes stems into lentinus edodes stem powder; adding the mushroom stem powder into 300mg/mL sodium hydroxide solution which is 5-200 times of the mushroom stem powder, boiling, keeping for 1-10 hours, and filtering through a 20-180-mesh sieve to obtain crude cellulose; adding 9.97mol/L hydrogen peroxide solution with 5-50 times volume of the crude cellulose into the crude cellulose, stirring the crude cellulose in a stirrer at 1000rpm for 1h at 30-70 ℃, stirring the crude cellulose at 25 ℃ for further stirring at 1000rpm for 1-24 h, sieving the obtained mixture with a 20-180-mesh sieve, and washing the obtained mixture with deionized water until the pH value is 7.0-7.9 to obtain NC; the NC is fibrous.
S2, preparing mushroom stems CNFs: homogenizing the lentinus edodes cellulose obtained in the step S1 for 2-20 times at a high pressure of 10-90 MPa to obtain CNFs; and (3) dissolving the CNFs in water to prepare a 10-25 mg/mL CNFs solution.
S3, preparing a colloidal emulsion: and (4) adding vegetable oil into the 10-25 mg/mL CNFs solution in the step (S2) according to the mass ratio of 1 (1-3), stirring to uniformly disperse the vegetable oil, and homogenizing for 2-20 times under 10-90 MPa to obtain the stable colloidal emulsion of the CNFs.
Preferably, the vegetable oil in step S3 is an edible oil such as soybean oil, salad oil, corn oil, sunflower seed oil or blend oil, and an aromatized vegetable oil obtained by extracting an edible oil as a base.
Example 1:
s1, extracting mushroom cellulose (NC): crushing dried shiitake stems with the water content of less than 13g/100g (national food safety standard edible fungi and products GB7096-2014 thereof), filtering through a 180-mesh sieve to obtain shiitake stem powder 50g, adding 250g of 300mg/mL sodium hydroxide solution, boiling for 1h, filtering the obtained mixture through a 20-mesh sieve to obtain crude cellulose, adding 200g of 9.97mol/L hydrogen peroxide solution for bleaching to obtain 40g of crude cellulose, stirring for 1h at 1000rpm in a stirrer at 30 ℃, stirring for 1h at 25 ℃ continuously at 1000rpm, filtering the obtained mixture through the 20-mesh sieve to obtain precipitate A, washing the precipitate A with deionized water to pH7.9, centrifuging for 60min at 1000rpm, and obtaining precipitate B to obtain shiitake cellulose (NC);
s2, preparing lentinus edodes stem Cellulose Nanofibers (CNFs): dissolving the lentinus edodes cellulose obtained in the step S1 in water to prepare a solution with the concentration of the lentinus edodes cellulose being 10mg/mL, and then homogenizing for 2 times under high pressure of 10MPa to obtain a Cellulose Nanofiber (CNFs) solution;
s3, preparing a colloidal emulsion: and (5) taking 50g of the CNFs solution obtained in the step S2, adding 100g of corn oil into the CNFs solution, stirring to uniformly disperse the corn oil, and homogenizing for 2 times under high pressure of 10MPa to obtain the colloidal emulsion with stable CNFs. The colloidal emulsion prepared in this example was homogeneous and stable, with no visible phase separation occurring after 30 days at room temperature (25 ℃).
Example 2:
s1, extracting mushroom cellulose (NC): crushing dried shiitake stems with the water content of less than 13g/100g (edible fungi and products thereof GB7096-2014 in national food safety standard), filtering the crushed shiitake stems through a 100-mesh sieve to obtain 50g of shiitake stem powder, adding 5000g of 300mg/mL sodium hydroxide solution, boiling the solution for 10h, filtering the obtained mixture through a 180-mesh sieve to obtain crude cellulose, adding 40g of the obtained crude cellulose into 200g of 9.97mol/L hydrogen peroxide solution, stirring the obtained mixture in a stirrer at 60 ℃ for 1h at 1000rpm, stirring the obtained mixture at 25 ℃ for 24h at 1000rpm, filtering the obtained mixture through a 180-mesh sieve to obtain precipitate A, washing the precipitate A with deionized water to pH7.9, and centrifuging the precipitate A at 5000rpm for 30min to obtain precipitate B to obtain shiitake cellulose (NC);
s2, preparing lentinus edodes stem Cellulose Nanofibers (CNFs): dissolving the lentinus edodes cellulose obtained in the step S1 in water to prepare a solution with the lentinus edodes cellulose concentration of 15mg/mL, and then homogenizing for 20 times under the high pressure of 90MPa to obtain Cellulose Nanofiber (CNFs) solution;
s3, preparing a colloidal emulsion: and (5) taking 50g of the CNFs solution obtained in the step S2, adding 150g of soybean oil into the CNFs solution, stirring to uniformly disperse the soybean oil, and homogenizing for 20 times under high pressure of 90MPa to obtain the colloidal emulsion with stable CNFs. The colloidal emulsion prepared in this example was homogeneous and stable, with no visible phase separation occurring after 30 days at room temperature (25 ℃).
Example 3:
s1, extracting mushroom cellulose (NC): crushing dried mushroom stems with the water content of less than 13g/100g (national food safety standard edible fungi and products thereof GB7096-2014), filtering the crushed dried mushroom stems through a 100-mesh sieve to obtain mushroom stem powder 10g, adding the mushroom stem powder into 2000g of 300mg/mL sodium hydroxide solution, boiling the mixture for 4 hours, filtering the obtained mixture through a 120-mesh sieve to obtain crude cellulose, adding 200g of 9.97mol/L hydrogen peroxide solution into the obtained crude cellulose, stirring the obtained mixture for 1 hour at 70 ℃ in a stirrer at 1000rpm, stirring the obtained mixture for 12 hours at 25 ℃ continuously at 1000rpm, filtering the obtained mixture through the 120-mesh sieve to obtain precipitate A, washing the precipitate A with deionized water to pH7.9, centrifuging the obtained mixture at 10000rpm for 5min, and obtaining precipitate B to obtain mushroom cellulose (NC);
s2, preparing lentinus edodes stem Cellulose Nanofibers (CNFs): dissolving the lentinus edodes cellulose obtained in the step S1 in water to prepare a solution with the lentinus edodes cellulose concentration of 20mg/mL, and then homogenizing for 10 times under the high pressure of 80MPa to obtain a CNFs solution;
s3, preparing a colloidal emulsion: and (5) taking 50g of the CNFs solution obtained in the step S2, adding 50g of soybean oil into the CNFs solution, stirring to uniformly disperse the soybean oil, and homogenizing for 10 times under high pressure of 80MPa to obtain the colloidal emulsion with stable CNFs. The colloidal emulsion prepared in this example was homogeneous and stable, and showed no visible phase separation after 30 days at room temperature (25 ℃).
Example 4:
s1, extracting mushroom cellulose (NC): crushing dried mushroom stems with the water content of less than 13g/100g (edible fungi and products thereof GB7096-2014) and filtering the crushed dried mushroom stems through a 20-mesh sieve to obtain mushroom stem powder 10g, adding 2000g of 300mg/mL sodium hydroxide solution into the mushroom stem powder, boiling the mixture for 4h, filtering the obtained mixture through a 120-mesh sieve to obtain crude cellulose, adding 200g of 9.97mol/L hydrogen peroxide solution into the obtained crude cellulose to bleach the obtained crude cellulose, stirring the obtained mixture for 1h at 70 ℃ in a stirrer at 1000rpm, stirring the obtained mixture for 12h at 25 ℃ at 1000rpm, filtering the obtained mixture through the 120-mesh sieve to obtain precipitate A, washing the precipitate A with water to pH7.9, and centrifuging the precipitate A at 5000rpm for 30min to obtain precipitate B to obtain the mushroom cellulose (NC);
s2, preparing lentinus edodes stem Cellulose Nanofibers (CNFs): dissolving the lentinus edodes cellulose obtained in the step S1 in water to prepare a solution with the lentinus edodes cellulose concentration of 25mg/mL, and then homogenizing for 10 times under the high pressure of 80MPa to obtain a CNFs solution;
s3, preparing a colloidal emulsion: and (5) taking 50g of the CNFs solution obtained in the step S2, adding 50g of soybean oil into the CNFs solution, stirring to uniformly disperse the soybean oil, and homogenizing for 10 times under high pressure of 80MPa to obtain the colloidal emulsion with stable CNFs. The colloidal emulsion prepared in this example was homogeneous and stable, and showed no visible phase separation after 30 days at room temperature (25 ℃).
Comparative example 1
S1, extracting mushroom cellulose (NC): crushing dried mushroom stems with the water content of less than 13g/100g (edible fungi and products thereof GB7096-2014) and filtering the crushed dried mushroom stems through a 200-mesh sieve to obtain mushroom stem powder 10g, adding 3000g of 300mg/mL sodium hydroxide solution into the mushroom stem powder, boiling the mixture for 12h, filtering the obtained mixture through the 200-mesh sieve to obtain crude cellulose, adding 1000g of 9.97mol/L hydrogen peroxide solution into the obtained crude cellulose 10g for bleaching, stirring the obtained crude cellulose in a stirrer at 1000rpm under the condition of 100 ℃ for 1h, stirring the obtained mixture at 25 ℃ for 48h at 1000rpm, filtering the obtained mixture through the 200-mesh sieve, taking precipitate A, washing the precipitate A with deionized water to pH7.9, centrifuging the precipitate A at 5000rpm for 30min, and taking precipitate B to obtain mushroom cellulose (NC);
s2, preparing lentinus edodes stem Cellulose Nanofibers (CNFs): dissolving the lentinus edodes cellulose obtained in the step S1 in water to prepare a solution with the lentinus edodes cellulose concentration of 5mg/mL, and then homogenizing for 30 times under 100MPa to obtain a CNFs solution;
s3, preparing a colloidal emulsion: and step S2, adding 200g of soybean oil into 50g of the CNFs solution, stirring to uniformly disperse the soybean oil, and homogenizing for 30 times under the high pressure of 100MPa to obtain the colloidal emulsion. This comparative example did not produce a stable emulsion and macroscopic phase separation occurred after high pressure homogenization.
Comparative example 2
S1, extracting mushroom cellulose (NC): crushing dried mushroom stems with the water content of less than 13g/100g (edible fungi and products thereof GB7096-2014) and filtering the crushed stems through a 200-mesh sieve to obtain mushroom stem powder 10g, adding 3000g of 300mg/mL sodium hydroxide solution into the mushroom stem powder, boiling the mixture for 30h, filtering the obtained mixture through the 200-mesh sieve to obtain crude cellulose, adding 10g of the obtained crude cellulose into 10g of 9.97mol/L hydrogen peroxide solution for bleaching, stirring the obtained mixture in a stirrer at 1000rpm for 1h at 4 ℃, stirring the obtained mixture at 1000rpm for 30min at 25 ℃, filtering the obtained mixture through the 200-mesh sieve to obtain precipitate A, washing the precipitate A with deionized water to pH7.0, and centrifuging the precipitate A at 5000rpm for 30min to obtain precipitate B, thus obtaining the mushroom cellulose (NC);
s2, preparing lentinus edodes stem Cellulose Nanofibers (CNFs): dissolving the lentinus edodes cellulose obtained in the step S1 in water to prepare a solution with the concentration of the lentinus edodes cellulose being 30mg/mL, and then homogenizing for 1 time under the high pressure of 5MPa to obtain a CNFs solution;
s3, preparing a colloidal emulsion: step S2, adding 50g of soybean oil into the CNFs solution, stirring to uniformly disperse the soybean oil, and homogenizing under 5MPa for 1 time to obtain colloidal emulsion. The emulsion prepared in this comparative example was very fluid and did not form a gel-like emulsion, with visible phase separation after 2 days at room temperature (25 ℃).
Comparative example 3
Dissolving commercially available plant cellulose nanocrystals (20161006, New chemical Co., Ltd., lake, Ling lake) in water to obtain 10mg/mL cellulose nanocrystal solution, adding 100g soybean oil into 100g cellulose nanocrystal solution with concentration of 10mg/mL, stirring to uniformly disperse the soybean oil, and homogenizing under 20MPa for 1 time to obtain emulsion. The emulsion prepared by the method has strong fluidity, can not become colloidal emulsion, and can be separated by naked eyes after being placed at room temperature (25 ℃) for 4 hours.
The appearance diagrams of the emulsions prepared in the examples are shown in fig. 1 to 7, the emulsions prepared in the examples 1 to 4 have remarkable colloidal emulsion characteristics, the emulsions are in a uniform and stable state, the fluidity is low, and the emulsions do not flow down when inverted. The emulsions in comparative examples 1-3 are in a flowing state, the emulsion in comparative example 1 is unstable, water is separated out at the lower layer, and oil drops at the top of the emulsion in comparative example 3 are clearly visible; in comparative example 2, however, the emulsion had a high fluidity and was not able to form a gel-like emulsion by flowing down slowly along the bottle wall.
FIGS. 8 to 13 are laser confocal scanning images of the emulsions prepared in examples 1 to 4 and comparative examples 1 to 2. The results show that the emulsions prepared in each example are in the form of oil-in-water emulsions, and the CNFs are dispersed in the continuous phase and coated on the surface of oil droplets, thereby achieving the effect of stabilizing the emulsions. The oil droplets in the colloidal emulsions of examples 1-4 all had a uniform circular profile, whereas the oil droplets became non-spherical in comparative examples 1 and 2 and were in a state of being coalesced. In comparative example 1, the oil droplets are highly dispersed and have a large diameter, at which time the oil droplets are aggregated into large particles, thereby decreasing the stability of the emulsion. Whereas in comparative example 2, the redundant CNFs intertwine in the continuous phase, these unevenly dispersed CNFs cause the oil droplets to coalesce and assume a polygonal shape, resulting in a decreased stability of the emulsion. The microscopic morphology of the emulsions shows that the emulsions prepared in examples 1-4 have the morphological characteristics of oil-in-water emulsions, and the CNFs are uniformly distributed on the surface of oil droplets, so that stable colloidal emulsions are formed. Whereas the emulsions prepared in comparative examples 1-2 exhibited an unstable morphology.
FIGS. 14 to 19 are scanning electron micrographs of the emulsions prepared in examples 1 to 4 and comparative examples 1 to 2, and the results further confirm the microstructure of the emulsion exhibited by the laser confocal coating. As can be seen, CNFs are interconnected in the emulsion to form a porous bridged network, so that oil droplets are embedded therein to stabilize the emulsion. This CNFs bridging network is less in comparative example 1, and thus the oil droplets in comparative example 1 exhibit an aggregated state, suggesting instability of the emulsion. The CNFs networks in the embodiments 1-4 are uniformly distributed on the continuous phase and the surface of oil drops, and the emulsion has high stability. In comparative example 2, however, it was observed that a large excess of CNFs was present in the continuous phase, and the excess CNFs twisted to form a compact small agglomerate, further confirming the results of confocal laser scanning. In addition, unlike the smooth oil droplet surfaces observed in examples 1-4 and comparative example 1, the excessive amount of CNFs agglomerates roughens the surface of the oil droplets in comparative example 2, and these agglomerates can form irregular steric hindrance, resulting in decreased emulsion stability.
FIG. 20 is a graph showing the measurement of the degree of lipid oxidation in emulsions prepared in examples 1 to 4 and comparative examples 1 to 2 by the thiobarbituric acid method. The thiobarbituric acid number (TBARS) of the soybean oil during 20 days of storage was between 1.26. + -. 0.17mmol/kg of oil and 2.84. + -. 0.14mmol/kg of oil, which was much higher than the TBARS of the emulsions of examples 1-4 and comparative examples 1-2 after storage for the same period of time. This result is probably because the CNFs coated on the surface of the oil droplets inhibited the oxidation of lipids, thereby reducing TBARS in the emulsion. Notably, the TBARS of comparative example 2 is significantly higher than example 4, which is a lower concentration of CNFs, a result that may be related to the instability of the emulsion of comparative example 2. The exposed oil droplets on the surface of comparative example 2 may exacerbate lipid oxidation of soybean oil, compared to comparative example 1 having an emulsifying layer above the water layer (see fig. 6).
FIG. 21 is a graph showing the emulsion index of the emulsions prepared in examples 1 to 4 and comparative examples 1 to 2 measured by centrifugation; wherein the emulsification index is calculated by the following formula, and the lower case letters a to e in the figure represent significant differences (p < 0.05).
As can be seen, the emulsification index between the examples is between 49.94 + -2.04% and 99.80 + -1.05%. Of these, comparative example 1 had the lowest emulsification index of 49.94 ± 2.04%. The colloidal emulsions prepared in examples 1 to 4 all exhibited a state of a uniform emulsion layer at the top and a precipitated water layer at the bottom after centrifugation. However, the emulsion of comparative example 2 has a higher emulsification index, but significant oil droplet separation is observed at the top, suggesting instability of the emulsion prepared in comparative example 2.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be able to cover the technical solutions and the inventive concepts of the present invention within the technical scope of the present invention.
Claims (5)
1. A method for preparing colloidal emulsion stabilized by cellulose nano-fibers is characterized by comprising the following steps:
s1, extracting the lentinus edodes cellulose: taking dried shiitake stipe powder, adding 300mg/mL sodium hydroxide solution with the weight 5-200 times of that of the dried shiitake stipe powder, boiling, keeping for 1-10 h, and filtering by a 20-180-mesh sieve to obtain a precipitate as crude cellulose; adding 9.97mol/L hydrogen peroxide solution which is 5-50 times of the weight of the crude cellulose into the crude cellulose, stirring the crude cellulose at 30-70 ℃ and 1000rpm for 1h, stirring the crude cellulose at 25 ℃ and 1000rpm for 1-24 h, sieving the obtained mixture by using a 20-180-mesh sieve, taking a precipitate A, washing the precipitate A with deionized water to a pH value of 7.0-7.9, and centrifuging the precipitate B to obtain the lentinan;
s2, preparing the lentinus edodes stem cellulose nano fiber: dissolving the lentinus edodes cellulose obtained in the step S1 in water to prepare a solution with the lentinus edodes cellulose concentration of 10-25 mg/mL, and homogenizing for 2-20 times under 10-90 MPa to obtain a cellulose nano-fiber solution;
s3, preparing a colloidal emulsion: taking the cellulose nanofiber solution with the concentration of 10-25 mg/mL in the step S2, adding vegetable oil, and homogenizing for 2-20 times under 10-90 MPa to obtain colloidal emulsion with stable cellulose nanofiber; wherein the weight ratio of the cellulose nanofiber solution to the vegetable oil is 1 (1-3).
2. The method for preparing cellulose nanofiber-stabilized colloidal emulsion according to claim 1, wherein the dried shiitake stem powder of step S1 is prepared by: crushing dried lentinus edodes stems into 20-180-mesh powder; the water content of the dried mushroom stems is less than 13g/100 g.
3. The method for preparing the colloidal emulsion stabilized by cellulose nanofibers according to claim 1, wherein the centrifugation parameters in step S1 are 1000-10000 rpm for 5-60 min.
4. The method of preparing a colloidal emulsion stabilized by cellulose nanofibers according to claim 1, wherein said vegetable oil of step S3 is soybean oil, salad oil, corn oil, sunflower seed oil or blend oil.
5. The method of preparing a cellulose nanofiber-stabilized colloidal emulsion according to claim 1, comprising the steps of:
s1, extracting the lentinus edodes cellulose: crushing dried lentinus edodes stems with the water content of less than 13g/100g, filtering through a 20-mesh sieve to obtain 10g of lentinus edodes stem powder, adding 2000g of 300mg/mL sodium hydroxide solution into the lentinus edodes stem powder, boiling for 4 hours, and filtering the obtained mixture through a 120-mesh sieve to obtain crude cellulose; adding 10g of the crude cellulose into 200g of 9.97mol/L hydrogen peroxide solution for bleaching, stirring at 70 ℃ and 1000rpm for 1h, stirring at 25 ℃ and 1000rpm for 12h, filtering the obtained mixture by using a 120-mesh sieve to obtain a precipitate A, washing the precipitate A by using deionized water until the pH value is 7.9, and centrifuging to obtain a precipitate B, thus obtaining the lentinus edodes cellulose;
s2, preparing mushroom stem cellulose nano-fibers: dissolving the lentinus edodes cellulose obtained in the step S1 in water to prepare a solution with the lentinus edodes cellulose concentration of 25mg/mL, and homogenizing for 10 times under 80MPa to obtain a cellulose nano-fiber solution;
s3, preparing a colloidal emulsion: and (5) taking 50g of the cellulose nanofiber solution obtained in the step (S2), adding 50g of soybean oil, and homogenizing for 10 times under 80MPa to obtain the colloidal emulsion with stable cellulose nanofiber.
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