CN111194767A - Litsea cubeba essential oil nanoemulsion - Google Patents
Litsea cubeba essential oil nanoemulsion Download PDFInfo
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- CN111194767A CN111194767A CN202010085098.0A CN202010085098A CN111194767A CN 111194767 A CN111194767 A CN 111194767A CN 202010085098 A CN202010085098 A CN 202010085098A CN 111194767 A CN111194767 A CN 111194767A
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/14—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12
- A23B4/18—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12 in the form of liquids or solids
- A23B4/20—Organic compounds; Microorganisms; Enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a litsea cubeba essential oil nanoemulsion which is prepared by compounding a water phase and an organic phase containing litsea cubeba essential oil, wherein the mass ratio of the litsea cubeba essential oil in the organic phase is (0.5-2): 1, and the mass ratio of the organic phase to the water phase is (0.15-0.25): 1. The litsea cubeba essential oil nanoemulsion solves the problem that litsea cubeba essential oil as an oily substance is not easy to dissolve in a water phase, has good low-temperature stability and strong antibacterial and antioxidant properties, can inhibit the decay and quality maintenance of aquatic products, prolongs the shelf life of the aquatic products, and has wide application prospect in the field of preservation and corrosion prevention in the process of cold and fresh aquatic products.
Description
Technical Field
The invention relates to litsea cubeba essential oil nanoemulsion.
Background
In the processes of processing, storing and circulating, food is decayed due to oxidation and propagation of harmful microorganisms, and in order to keep the nutritional value and the quality of the food, the traditional aquatic product preservative and fresh-keeping mode comprises physical processing and chemical synthesis of bacteriostatic agents, but potential safety hazards exist. Wherein, the physical processing technology has high equipment cost and is easy to cause the damage of food nutrient components; chemical bacteriostatic agents such as chemical compounds and antibiotics are easy to cause potential safety hazards such as bacterial drug resistance and the like, and potential harm is caused to human health when too much chemical bacteriostatic agent is ingested.
The plant essential oil is highly concentrated extract of aromatic plant, and can be obtained from flower, leaf, stem, root, fruit, seed, bark, etc. of herbaceous plant by squeezing, distilling, etc. Different from the traditional chemical preservative, the plant essential oil belongs to a natural antibacterial agent and has stronger antibacterial, bacteriostatic and antioxidant effects, so the plant essential oil is often applied to the field of food storage and preservation and serves as a substitute of a chemical compound and antibiotics. On the other hand, the volatility and strong hydrophobicity of the plant essential oil directly limit the application of the plant essential oil as a food additive in food; in addition, hydrophobic groups in plant essential oils may combine with proteins and lipids to form an uneven distribution, reducing antibacterial efficacy, and thus the effect in practical use is not ideal.
Disclosure of Invention
The invention aims to provide the litsea cubeba essential oil nanoemulsion, which solves the problem that litsea cubeba essential oil as an oily substance is not easy to dissolve in a water phase, has good low-temperature stability and strong antibacterial and antioxidant properties, can inhibit the putrefaction and quality maintenance of aquatic products, prolongs the shelf life of the aquatic products, and has wide application prospect in the field of preservation and corrosion prevention in the process of cold and fresh aquatic products.
In order to achieve the purpose, the invention adopts the following technical scheme:
the litsea cubeba essential oil nanoemulsion is prepared by compounding a water phase and an organic phase containing litsea cubeba essential oil, wherein the mass ratio of the litsea cubeba essential oil in the organic phase is (0.5-2) to 1, and the mass ratio of the organic phase to the water phase is (0.15-0.25) to 1.
The organic phase contains medium chain triglycerides which form a homogeneous organic phase with litsea cubeba essential oil.
The mixing mass ratio of the medium chain triglyceride to the litsea cubeba essential oil is (0.5-2): 1.
The litsea cubeba essential oil nanoemulsion also comprises an emulsifier.
Tween-80 is used as emulsifier.
The mass ratio of the tween-80 in the formed litsea cubeba essential oil nanoemulsion is as follows: 4 to 8 percent.
The water phase is distilled water or phosphate buffer.
And a cosurfactant glycerol is also added, and the mass ratio of the glycerol in the formed litsea cubeba essential oil nanoemulsion is as follows: 0.5 to 1.5 percent.
The litsea cubeba essential oil nanoemulsion comprises 15% of an organic phase, 78% of a water phase, 6% of tween-80 and 1% of glycerol by mass ratio, wherein the organic phase is a uniform organic phase formed by medium chain triglyceride and litsea cubeba essential oil, the medium chain triglyceride is 2:1, and the water phase is 0.1mM of phosphate buffer.
Compared with the prior art, the invention has the beneficial effects that: the application solves the problem that litsea cubeba essential oil is not easy to dissolve in a water phase as an oily substance, the litsea cubeba essential oil can be applied to seawater products in a smearing or adding mode after being wrapped in water-based nano-milk, the stability of the litsea cubeba essential oil at low temperature is also beneficial to the application of aquatic products in a cold chain process, the strong antibacterial and antioxidant properties of the litsea cubeba essential oil can also cooperate with low-temperature treatment to inhibit the putrefaction and quality maintenance of the water products, and the shelf life of the aquatic products is prolonged, so that the litsea cubeba essential oil has a wide application prospect in the field.
The application adopts a phosphate buffer solution water phase, and the litsea cubeba essential oil forms an oil-in-water nanoemulsion, has a whitish appearance, and has the following advantages: (1) the litsea cubeba oil-in-water nanoemulsion has moderate viscosity and stable property, and can be infinitely diluted by a phosphate buffer; (2) the litsea cubeba essential oil wrapped by the nano-emulsion has strong low-temperature stability, combines strong antibacterial property and active oxygen scavenging capacity, and can be applied to preservation and preservation of aquatic products and quality maintenance in a matching way at low temperature; (3) the litsea cubeba essential oil wrapped by the nanoemulsion has the effects of simultaneously inhibiting putrefying bacteria and pathogenic bacteria and simultaneously inhibiting the formation of cell envelopes of the putrefying bacteria and the pathogenic bacteria, so that the drug resistance of harmful microorganisms can be reduced.
The glycerol has low cost, easy industrial acquisition and nutritional function. The application adopts 1% of glycerol to form an organic phase together with the litsea cubeba essential oil, reduces the cost of independently using the litsea cubeba essential oil as the organic phase, and improves the practical applicability and value of the product.
The aquatic product spoilage bacteria are Shewanella borrelia, and the pathogenic bacteria are Listeria monocytogenes. The litsea cubeba nanoemulsion provided by the embodiment of the invention is very suitable for inhibiting the growth of putrefying bacteria and pathogenic bacteria in aquatic products and the formation of biofilm.
Drawings
Table 1 orthogonal analysis of nanoemulsion essential oil for preservation and freshness of aquatic products;
table 2 antibacterial property analysis of litsea cubeba pure essential oil;
FIG. 1 is a particle size distribution diagram, a microscopic view and an actual view of a Litsea cubeba nanoemulsion according to an embodiment of the invention;
FIG. 2 shows the particle size change (stability) of the Litsea cubeba nanoemulsion at 4 deg.C, 25 deg.C and 40 deg.C according to the embodiment of the present invention;
FIG. 3 is a graph showing the growth inhibitory effect of the Litsea cubeba nanoemulsion on aquatic product spoilage bacteria (Shewanella borealis) and pathogenic bacteria (Listeria monocytogenes) according to an embodiment of the present invention;
FIG. 4 is a graph showing the effect of the litsea cubeba nanoemulsion on biofilm inhibition by aquatic product spoilage bacteria (Shewanella borrelia) and pathogenic bacteria (Listeria monocytogenes) according to the embodiment of the invention.
FIG. 5 is a graph showing the effect of a Litsea cubeba nanoemulsion on the scavenging ability of active oxygen ions according to an embodiment of the present invention;
FIG. 6 shows the content of volatile basic nitrogen (mg/100g) in fish meat;
FIG. 7 shows the effect of the litsea cubeba essential oil nanoemulsion on the total bacterial count (TVC value) of refrigerated large yellow croaker at 4 ℃.
Detailed Description
In order to make the technical solution of the present invention clearer, the present invention will be described in detail below with reference to fig. 1 to 7. It should be understood that the detailed description and specific examples, while indicating the scope of the invention, are intended for purposes of illustration only and are not intended to limit the scope of the invention.
Example 1
The litsea cubeba essential oil nanoemulsion is prepared by compounding an aqueous phase and an organic phase containing litsea cubeba essential oil, wherein the mass ratio of the litsea cubeba essential oil in the organic phase is 0.5:1, the mass ratio of the organic phase to the aqueous phase is 0.15:1, and the aqueous phase is distilled water or a phosphate buffer solution.
Preferably, the organic phase contains medium chain triglyceride, the medium chain triglyceride and the litsea cubeba essential oil form a uniform organic phase, and the mixing mass ratio of the medium chain triglyceride to the litsea cubeba essential oil is 0.5: 1.
The litsea cubeba essential oil nanoemulsion also comprises an emulsifier and a cosurfactant, wherein the emulsifier is tween-80, the tween-80 accounts for 4% of the mass of the litsea cubeba essential oil nanoemulsion, the cosurfactant is glycerol, and the mass ratio of the glycerol in the formed litsea cubeba essential oil nanoemulsion is as follows: 0.5 percent.
Example 2
The litsea cubeba essential oil nanoemulsion is prepared by compounding an aqueous phase and an organic phase containing litsea cubeba essential oil, wherein the mass ratio of the litsea cubeba essential oil in the organic phase is 1:1, the mass ratio of the organic phase to the aqueous phase is 0.2:1, and the aqueous phase is distilled water or a phosphoric acid buffer solution.
Preferably, the organic phase contains medium chain triglyceride, the medium chain triglyceride and the litsea cubeba essential oil form a uniform organic phase, and the mixing mass ratio of the medium chain triglyceride to the litsea cubeba essential oil is 0.2: 1.
The litsea cubeba essential oil nanoemulsion also comprises an emulsifier and a cosurfactant, wherein the emulsifier is tween-80, the tween-80 accounts for 6% of the mass of the litsea cubeba essential oil nanoemulsion, the cosurfactant is glycerol, and the mass ratio of the glycerol in the formed litsea cubeba essential oil nanoemulsion is as follows: 1 percent.
Example 3
The litsea cubeba essential oil nanoemulsion is prepared by compounding an aqueous phase and an organic phase containing litsea cubeba essential oil, wherein the mass ratio of the litsea cubeba essential oil in the organic phase is 2:1, the mass ratio of the organic phase to the aqueous phase is 0.25:1, and the aqueous phase is distilled water or a phosphoric acid buffer solution.
Preferably, the organic phase contains medium chain triglyceride, the medium chain triglyceride and the litsea cubeba essential oil form a uniform organic phase, and the mixing mass ratio of the medium chain triglyceride to the litsea cubeba essential oil is 2: 1.
The litsea cubeba essential oil nanoemulsion also comprises an emulsifier and a cosurfactant, wherein the emulsifier is tween-80, the tween-80 accounts for 8% of the mass of the litsea cubeba essential oil nanoemulsion, the cosurfactant is glycerol, and the mass ratio of the glycerol in the formed litsea cubeba essential oil nanoemulsion is as follows: 1.5 percent.
Example 4
A Litsea cubeba essential oil nanoemulsion is prepared by compounding an aqueous phase and an organic phase containing Litsea cubeba essential oil, wherein the mass ratio of the organic phase is 22%, and the mass ratio of the aqueous phase is 78%; wherein the water phase is 0.1mM phosphate buffer solution, the organic phase comprises litsea cubeba essential oil, medium chain triglyceride, tween-80 and glycerol, the mass ratio of the litsea cubeba essential oil is 10%, the mass ratio of the medium chain triglyceride is 5%, the mass ratio of the tween-80 is 6% and the mass ratio of the glycerol is 1%.
Example 5
The preparation method of the litsea cubeba essential oil nanoemulsion comprises the following steps:
the method comprises the following steps: respectively weighing an organic phase and a water phase according to the mass ratio, and slowly adding the water phase into the organic phase;
step two: putting the mixed solution into a stirrer, wherein the stirring speed is 800 revolutions per minute, and stirring for 30 minutes;
step three: then, homogenizing for 5 minutes at 12000 rpm by using a high-speed homogenizer;
step four: and finally, dispersing the solution by adopting ultrasound to obtain the litsea cubeba essential oil nanoemulsion.
Preferably, the ultrasonic power is 940W/100ml of solution, and the ultrasonic time is 8-16 minutes.
Such as tween-80, an emulsifier and/or glycerol, a co-surfactant, may be weighed in step one as part of the organic phase.
Example 6: influence of organic phase composition on litsea cubeba essential oil nanoemulsion effect
In this example, the aqueous phase was selected from 0.1mM phosphate buffer (abbreviated as PBS, pH7.0), and the organic phase consisted of: litsea cubeba essential oil, medium chain triglyceride, tween-80 and glycerol. The influence effect of different addition ratios on the particle size of the nanoemulsion is selected, and the particle size is measured by a dynamic light scattering method, which is specifically shown in table 1 and fig. 1.
Table 1. nanoemulsion orthogonal experiment (n ═ 3)a
When using varying concentrations of litsea cubeba essential oil, medium chain triglyceride, tween-80, glycerol as the organic phase, emulsions of different average particle size can be formed (table 1). When the litsea cubeba essential oil (the litsea cubeba essential oil: medium chain triglyceride is 2:1 (mass ratio)) is contained in 15%, tween-80 is 6%, glycerol is 1% and PBS is 78%, the optimal nano particle size (101.2 +/-0.1 nm, shown in figures 1-A, B and C) can be effectively obtained, and the litsea cubeba essential oil nanoemulsion is prepared according to the component ratio in the following examples.
Example 7: experiment for influence of stability of litsea cubeba essential oil nanoemulsion at different temperatures
The litsea cubeba essential oil nanoemulsion used for preservation and freshness of aquatic products in the embodiment is the litsea cubeba essential oil nanoemulsion of the embodiment 4.
The embodiment provides the stability of the litsea cubeba essential oil nanoemulsion at different temperatures. Standing fructus Litseae nanoemulsion at 4 deg.C, 25 deg.C, and 40 deg.C for 30 days, and measuring average particle diameter every 10 days. The increase of the temperature can cause the changes of the emulsion such as the enhancement of Brownian motion and the like, thereby causing the phenomena of the physical stability reduction, the milk separation and the like of the litsea cubeba nano emulsion. Therefore, the key point of the nano emulsion in the fields of food and medicine is to maintain good storage stability at a certain temperature. The litsea cubeba essential oil nanoemulsion is stored for 30 days at the temperature of 4 ℃, 25 ℃ and 40 ℃, and the average particle size of the litsea cubeba essential oil nanoemulsion increases to different degrees along with the increase of the temperature and the storage time. The average particle size increased from 101.1nm to 374.23nm after 30 days of storage at 40 ℃. It is worth noting that the litsea cubeba essential oil nanoemulsion in the embodiment can keep good stability when stored at normal temperature (25 ℃) and low temperature (4 ℃) for 30 days, and therefore is suitable for preservation and fresh keeping of aquatic products during ice fresh refrigeration. Fig. 2 shows the particle size change (stability) of the litsea cubeba nanoemulsion at 4 ℃, 25 ℃ and 40 ℃ according to an embodiment of the invention.
Example 8: antibacterial property analysis of litsea cubeba essential oil and nanoemulsion thereof
The litsea cubeba essential oil nanoemulsion used for preservation and freshness of aquatic products in the embodiment is the litsea cubeba essential oil nanoemulsion of the embodiment 4. Preparing the litsea cubeba essential oil nanoemulsion into a series of twice-diluted gradient concentrations, mixing the litsea cubeba essential oil nanoemulsion with an LB culture medium which is melted and cooled to about 50 ℃ to prepare a litsea cubeba nanoemulsion with the mass fraction of 0.125-8%, and pouring the litsea cubeba essential oil nanoemulsion to prepare a mixed flat plate. 5 μ L of the prepared bacterial suspension was transferred and inoculated onto the surface of the plate with an initial bacterial concentration of about 104cfu/mL. And (3) taking an LB blank plate and the nanoemulsion without adding litsea cubeba essential oil as a control, culturing the test plate in a constant temperature incubator at 30 ℃ for 24 hours, and observing whether bacteria grow.
The growth test adopts a detection method adopting a live bacterium CTC-flow cytometer: taking the bacterial liquid cultured for the corresponding time, centrifuging at 12000r/min for 10min, removing supernatant, and resuspending the bacterial liquid with 1mL sterile PBS or normal saline. The total colony number measuring method of the CTC-flow cytometer comprises the following steps: mu.L of the sample was added to a 96-well plate, 4. mu.L of 50mM CTC was added, and the plate was incubated at 37 ℃ in the dark for 3 hours, followed by counting using a flow cytometer. A200. mu.L sample was added to a 96-well plate without CTC as negative. Flow cytometer parameters were set to: FL1 was 4.11, FL2 was 6.29, FL3 was 5.47, and the volume was 50 μ L per measurement. The optimal reaction conditions for CTC staining were: the final concentration of CTC is 2mM, the incubation is carried out for 3h at 37 ℃ in the dark, and the minimum detection limit of the method is 103CFU/mL. As a result, as shown in FIG. 3, the viable cell count of Listeria monocytogenes and Shewanella borreliana increased to 10 days after the addition of any of the treated control groups9cfu/mL, the litsea cubeba essential oil can obviously reduce the viable count of two bacteria, and shows the initial sterilization effect and the whole bacteriostasis effect after being wrapped by the nano-emulsion.
TABLE 2 antibacterial property of Litsea cubeba essential oil
Note: "+" indicates that there is bacterial growth; "-" indicates no bacterial growth.
FIG. 3 is a graph showing the growth inhibition effect of the litsea cubeba nanoemulsion on aquatic product spoilage bacteria (Shewanella borrelia) and pathogenic bacteria (Listeria monocytogenes) according to the embodiment of the invention
Example 9: analysis of inhibition effect of litsea cubeba essential oil and nanoemulsion thereof on biological envelope
The litsea cubeba essential oil nanoemulsion used for preservation and freshness of aquatic products in the embodiment is the litsea cubeba essential oil nanoemulsion of the embodiment 4. The biofilm test adopts a crystal violet detection method: the activated strain was diluted 1:100 and inoculated into a 96-well cell plate at a volume of 100. mu.L/well (the inoculum size was adjusted appropriately according to the incubation time, and if the incubation time exceeded two days, the inoculum size was increased appropriately to prevent air drying of the inoculum). Then putting the sample into the culture temperature for constant temperature culture at 4 ℃. Considering that different strains have different periods of pellicle formation and exfoliation at different culture temperatures, it is suggested that measurement can be selected every 12h or every 24h to find the peak of pellicle formation by the strain when cultured at the appropriate temperature. After the culture time is up, the excess medium is aspirated by a pipette gun, and the cells are washed 2 times with 150. mu.L of ultrapure water, the last wash being as complete as possible, in order to shorten the subsequent drying time, and dried or baked. Adding 150 mu L of crystal violet dye solution, standing for 20min, and sucking away the dye solution. Washing with 200 μ L water twice, air drying, adding 200ul 95% ethanol, decolorizing for 5min, placing 100 μ L per well in a new 96-well plate, and reading OD value at 595nm with an enzyme labeling instrument. The result is shown in fig. 4, the pure litsea cubeba essential oil has the effect of inhibiting the formation of mycoderm, and the inhibiting effect is obviously improved by adding the litsea cubeba nanoemulsion essential oil.
Example 10
The litsea cubeba essential oil nanoemulsion used for preservation and freshness of aquatic products in the embodiment is the litsea cubeba essential oil nanoemulsion of the embodiment 4.
DPPH free radical clearance determination samples of methanol solutions with mass concentrations of 4,8,10,16,24,32 and 50g/L were prepared, 200. mu.L and 4mL samples of 6X 10-5mixing mol/L DPPH methanol solution, measuring absorbance at the wavelength of 517nm, and taking DPPH methanol solution of pure litsea cubeba essential oil sample as blank sample. The number of replicates was 3 and the radical clearance was performed as follows:
free radical scavenging rate (%) - (A)Sample 520nm-ABlank 520nm)/(ASample 520nm-ABlank 520nm)]×100
Wherein A isSample 520nmIs the absorbance of the sample at 520 nm; a. theBlank 520nmIs the absorbance at 520nm of the blank.
In conclusion, the litsea cubeba essential oil nanoemulsion has low-temperature stability, can inhibit the growth of bacteria by spoilage bacteria and pathogenic bacteria of aquatic products, has a remarkable inhibition effect on biological envelopes, and can be applied to preservation and preservation of aquatic products and quality maintenance in cooperation with low temperature in combination with strong active oxygen scavenging capacity. FIG. 5 is a graph showing the effect of a Litsea cubeba nanoemulsion on the scavenging ability of active oxygen ions according to an embodiment of the present invention;
example 11: application of litsea cubeba essential oil nanoemulsion in fresh-keeping and corrosion prevention of salmon fillet
The litsea cubeba essential oil nanoemulsion used for preservation and freshness of aquatic products in the embodiment is the litsea cubeba essential oil nanoemulsion of the embodiment 4.
The embodiment provides an application of litsea cubeba essential oil nanoemulsion in chilled preservation of aquatic products. The method comprises the steps of purchasing a commercially available fresh salmon, flushing the surface of a raw material fish body with 5% saline water, washing off mucus and part of microorganisms, stripping fish skin on a super clean workbench, cutting muscles into sheets with uniform size (about 5cm multiplied by 5cm), immersing a fish sheet sample into litsea cubeba essential oil nano-emulsion and pure nano-emulsion to enable the surface of the fish sheet sample to be rapidly wrapped with a layer of preservative film, airing at room temperature for 5 minutes, and storing the coated salmon in a refrigerator at 4 ℃.
The pure nano-emulsion is prepared by compounding a water phase and an organic phase, wherein a phosphoric acid buffer solution is used for replacing litsea cubeba essential oil, the mass ratio of the organic phase is 12%, and the mass ratio of the water phase is 88%; wherein the water phase is 0.1mM phosphate buffer solution, the organic phase is composed of medium chain triglyceride, Tween-80 and glycerol, the mass ratio of the medium chain triglyceride is 5%, the mass ratio of the Tween-80 is 6%, and the mass ratio of the glycerol is 1%, and the preparation steps are as follows:
the method comprises the following steps: respectively weighing an organic phase and a water phase according to a preset mass ratio, and slowly adding the water phase into the organic phase;
step two: putting the mixed solution into a stirrer, wherein the stirring speed is 800 revolutions per minute, and stirring for 30 minutes;
step three: then, homogenizing for 5 minutes at 12000 rpm by using a high-speed homogenizer;
step four: finally, dispersing the solution by adopting ultrasound to obtain the pure nano-emulsion.
Volatile basic nitrogen measurements were performed on salmon while in storage, and the results shown in fig. 6 were obtained, and the data are the average values after 3 measurements. Method for measuring volatile basic nitrogen: mashing the fish sample by using a tissue mashing machine, weighing 5g of the mashed fish sample into a conical flask, adding 45mL of perchloric acid solution, homogenizing for 2min, filtering by using filter paper, storing the liquid to be detected at 4 ℃, and determining by using a detection method by referring to national standard TVB-N. The total number of colonies (TVC value s) was determined according to the method of national Standard GB 4789.2-2016, and counted by plate pour method.
As can be seen from fig. 6, compared with the control group (Blank), the content of volatile basic nitrogen in the fish meat after the litsea cubeba essential oil nanoemulsion is added is obviously lower, the standard of secondary freshness is that the volatile basic nitrogen is less than 30, and the freshness date after the litsea cubeba essential oil nanoemulsion is added is longer than the proportion, which shows that the method of the present invention has good freshness preservation and corrosion prevention effects.
The results of colony count are shown in FIG. 7, compared with the control group (Blank) without any treatment, the colony count of salmon fillet storage 4d in the Blank control group (Blank) and the pure nanoemulsion treatment group reaches 7.6log cfu/g and 6.4log cfu/g with the extension of storage time, which exceed the national specified sanitary standard; after the preservative fresh-keeping coating film of the aquatic product, namely the litsea cubeba essential oil nanoemulsion, is added, the bacterial growth is obviously inhibited, and the shelf life of the salmon fillet can be prolonged by more than 6d (the TVC value is 5.98log cfu/g).
Claims (9)
1. The litsea cubeba essential oil nanoemulsion is characterized in that: the litsea cubeba oil-containing emulsion is prepared by compounding a water phase and an organic phase containing litsea cubeba essential oil, wherein the mass ratio of the litsea cubeba essential oil in the organic phase is (0.5-2) to 1, and the mass ratio of the organic phase to the water phase is (0.15-0.25) to 1.
2. The litsea cubeba essential oil nanoemulsion as claimed in claim 1, characterized in that: the organic phase contains medium chain triglycerides which form a homogeneous organic phase with litsea cubeba essential oil.
3. The litsea cubeba essential oil nanoemulsion as claimed in claim 2, characterized in that: the mixing mass ratio of the medium chain triglyceride to the litsea cubeba essential oil is (0.5-2): 1.
4. The litsea cubeba essential oil nanoemulsion as claimed in claim 3, characterized in that: the litsea cubeba essential oil nanoemulsion also comprises an emulsifier.
5. The litsea cubeba essential oil nanoemulsion according to claim 4, characterized in that: tween-80 is used as emulsifier.
6. The litsea cubeba essential oil nanoemulsion as claimed in claim 5, characterized in that: the mass ratio of the tween-80 in the formed litsea cubeba essential oil nanoemulsion is as follows: 4 to 8 percent.
7. The litsea cubeba essential oil nanoemulsion as claimed in any one of claims 1 to 6, characterized in that: the water phase is distilled water or phosphate buffer.
8. The litsea cubeba essential oil nanoemulsion as claimed in claim 7, characterized in that: and a cosurfactant glycerol is also added, and the mass ratio of the glycerol in the formed litsea cubeba essential oil nanoemulsion is as follows: 0.5 to 1.5 percent.
9. The litsea cubeba essential oil nanoemulsion as claimed in claim 8, characterized in that: the preparation method comprises 15% of an organic phase, 78% of a water phase, 6% of tween-80 and 1% of glycerol by mass ratio, wherein the organic phase is a uniform organic phase formed by medium-chain triglyceride and litsea cubeba essential oil, the ratio of the medium-chain triglyceride to the litsea cubeba essential oil is 2:1, and the water phase is 0.1mM of phosphate buffer.
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