CN111449123A - Application of Ensifeng tea ethanol extract in inhibiting putrefaction of high-temperature cooked fish meat - Google Patents
Application of Ensifeng tea ethanol extract in inhibiting putrefaction of high-temperature cooked fish meat Download PDFInfo
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Abstract
The invention relates to an application of an Ensifera indica alcohol extract in inhibiting putrefaction of high-temperature cooked fish, which is characterized in that: the application of the Ensifera indica ethanol extract in inhibiting putrefaction of high-temperature cooked fish meat. The ampelopsis grossedentata ethanol extract has a simple extraction process, contains various active ingredients, is homologous in medicine and food, is green and healthy, can be used as a pure natural food preservative, and provides an important new product for prolonging the storage time of a fish product, improving the nutritional value of the fish product and improving the edible safety.
Description
Technical Field
The invention relates to an application of an Ensifera indica alcohol extract in inhibiting putrefaction of high-temperature cooked fish.
Background
Ampelopsis grossedentata (Ampelopsis grossedentata) is known as Ampelopsis grossedentata and commonly called white tea, mountain sweet tea, Cantonese tea, Longxu tea, etc. Is a plant of ampelopsis, and is a traditional medicinal and edible drink with the functions of tea and medicine. Vine tea is mainly prepared from leaves, has a small amount of stems and branches, and is also called Ampelopsis grossedentata tea, nectar tea and the like when being used as a medicine or drunk. In recent years, the chemical component analysis of ampelopsis grossedentata by scholars at home and abroad finds that the ampelopsis grossedentata is a special natural plant rich in flavonoid compounds and also rich in various nutritional components. The flavonoids are the main components of Ampelopsis grossedentata, the content of the flavonoids is richer than that of other plants, and the effective components of the flavonoids are ampelopsin, dihydromyricetin and the like. The content of the total flavone in the tender stems and leaves of the vine tea is up to 43.4 to 45.52 percent. The vine tea contains various flavonoid compounds, including dihydromyricetin, quercetin, taxifolin, kaempferol, vine tea extract, myricitrin, hesperetin, apigenin, etc., wherein the dihydromyricetin is the main active component in the vine tea. Vine tea has sweet taste, and has effects of clearing heat and detoxicating, and can be used for treating icterohepatitis, common cold wind-heat, sore throat, ulcerative equi, tinea pedis eczema, etc. The vine tea has high flavonoid content in plant, and its main flavone component is dihydromyricetin, and has effects of resisting oxidation, lowering blood pressure and regulating immunity. The ethanol crude extract (VTE) of Ampelopsis grossedentata has various physiological effects of inhibiting bacteria, resisting oxidation, diminishing inflammation, protecting liver, resisting tumor, resisting tyrosinase, resisting diabetes, reducing blood lipid, etc.
In the current food preservative application, chemical preservatives are mainly used. Chemical preservatives such as sulfur dioxide, sulfite and nitrite have accumulative toxicity to human bodies, and phenomena such as abuse, misuse and overproof use of the chemical preservatives occur occasionally, so that the chemical preservatives greatly threaten human health and cause a plurality of food safety problems. The natural preservative is non-toxic and harmless to human bodies, and also has certain nutritional value. The natural edible spice plants of perilla, bay, mustard seed, fennel and cinnamon are used as folk main natural preservatives for meat storage and preservation. Traditional Chinese medicinal materials become a new research hotspot of natural food preservatives, and forsythia extract of Oleaceae, plum extract of Rosaceae, artemisia selengensis extract of Compositae, rhubarb extract of Polygonaceae and the like are used for food preservative research.
The fish meat has high protein content, and is easy to cause the food to decay and deteriorate, thereby causing the economic loss of related food enterprises. The invention takes the ampelopsis grossedentata ethanol extract with various physiological effects as an object, researches the anticorrosion effect of the ampelopsis grossedentata ethanol extract serving as a pure natural food preservative on high-temperature cooked fish, and provides important references for prolonging the storage time of fish products, improving the nutritive value of the fish products and researching the edible safety.
The natural preservative applied to fish preservation at present mainly comprises lactic acid bacteria, tea polyphenol and chitosan film. The lactobacillus biological preservative developed by utilizing the growth competitive power of the lactobacillus has good preservation effect on the yellow croaker flesh, the sea bass blocks and the tilapia; the tea polyphenols have antioxidant effect, and can effectively slow down chain reaction and fish protein oxidation in the fat oxidation process of the chub meat blocks, and prolong the preservation time of the chub blocks; the edible film of chitosan can inhibit the growth of mesophilic and psychrophilic aerobic putrefying bacteria of Lateolabrax japonicus meat, and prolong the shelf life of Lateolabrax japonicus meat by about 20 days. The lactobacillus is extremely susceptible to the environment as a microorganism, and the metabolite of the lactobacillus can influence the flavor and the quality of fish meat, so that the application of the lactobacillus in the preservation of aquatic products is limited. The extraction process of the tea polyphenol and the chitosan-based membrane has higher requirements, and the large-scale application of the chitosan-based membrane in fish preservation is limited.
Disclosure of Invention
The invention aims to provide application of an Ensifera indica Gaertn tea ethanol extract in inhibiting putrefaction of high-temperature cooked fish.
The application of the Ensifeng tea ethanol extract in inhibiting the putrefaction of high-temperature cooked fish meat is characterized in that: the application of the Ensifera indica ethanol extract in inhibiting putrefaction of high-temperature cooked fish meat.
The application of the Ensifeng tea ethanol extract in inhibiting the putrefaction of high-temperature cooked fish meat is characterized in that: pulverizing dried Ampelopsis grossedentata, and soaking in 50% -70% ethanol at a material-liquid ratio of 1:40-55 for 17-19 h. Ultrasonic leaching at power of 190-210 w for 28-32 min, and concentrating under reduced pressure: concentrating the vine tea ethanol extract into pasty solid under the pressure of-0.085 MPa to-0.095 MPa and at the temperature of 35 ℃ to 40 ℃, and obtaining the vine tea ethanol extract preservative solution without ethanol solvent, wherein no liquid drop exists in a condensation tube of a rotary evaporator.
The application of the Ensify ampelopsis grossedentata ethanol extract in inhibiting putrefaction of fish cooked at high temperature is characterized in that the ampelopsis grossedentata ethanol extract is dissolved by sterile water to ensure that the concentration of the ampelopsis grossedentata ethanol extract is 40.0 mg.m L-1-50.0mg·mL-1Soaking cooked fish meat in water at a concentration of 40.0mg · m L-1-50.0mg·mL-1Soaking in the preservative solution for 30-35 min.
The application of the Ensify ampelopsis grossedentata ethanol extract in inhibiting putrefaction of fish cooked at high temperature is characterized in that the ampelopsis grossedentata ethanol extract is dissolved by sterile water to ensure that the concentration of the ampelopsis grossedentata ethanol extract is 45 mg.m L-1Soaking cooked fish meat in water at a concentration of 45.0mg · m L-1Soaking in the antiseptic solution for 33 min.
The anti-corrosion effect of the Engratia excelsa ethanol extract on high-temperature cooked fish is realized, bacteria causing high-temperature cooked fish spoilage are separated and identified by adopting a dilution plate method and a 16SRNA sequencing technology, the antibacterial activity of the Engratia excelsa ethanol extract on the spoilage bacteria is tested by using a filter paper sheet method, the antibacterial mechanism is researched by measuring the influence of the Engratia excelsa ethanol extract on the growth curve, the electric conductivity and the content of macromolecular dissolved substances of the spoilage bacteria, and the research result shows that the concentration of the Engratia excelsa ethanol extract is 30 mg.m L in the storage period of 15d-1The Engratia japonica ethanol extract can completely inhibit the growth of bacteria in high-temperature cooked fish, 2 Bacillus methylotrophicus T-10 and Alcaligenes faecalis T-9 which cause the high-temperature cooked fish to go bad are separated and identified, the Engratia japonica ethanol extract has medium-sensitive antibacterial activity (the diameter of an antibacterial ring is 10mm-20mm) on 2 putrefying bacteria, and the Minimum Inhibitory Concentration (MIC) of the Engratia japonica ethanol extract is 18.5mg m L-1、19.2mg·mL-1(ii) a The Endocarpus asiaticus ethanol extract plays a role in bacteriostasis by destroying the integrity of a thallus cell membrane and causing macromolecular substances to overflow.
The application of the Ensifera indica ethanol extract in inhibiting putrefaction of high-temperature cooked fish meat, provided by the invention, comprises multiple effective active ingredients: flavones, phenols, polysaccharides, volatile oil, amino acids, trace elements and the like, wherein the content of the flavones is 43.40-45.52%, and the main effective component dihydromyricetin in the flavones compound has the bacteriostatic action on staphylococcus aureus, multi-resistant pseudomonas aeruginosa, escherichia coli and bacillus subtilis and also has the antioxidant function. It is presumed that flavonoids such as dihydromyricetin in the ampelopsis grossedentata ethanol extract inhibits the reproduction of bacillus and alcaligenes faecalis in the high-temperature cooked fish meat, and can effectively slow down the chain reaction in the oxidation process of the fat in the fish meat blocks and the oxidation of the fish protein. The ampelopsis grossedentata ethanol extract has a simple extraction process, contains various active ingredients, is homologous in medicine and food, is green and healthy, can be used as a pure natural food preservative, and provides an important new product for prolonging the storage time of a fish product, improving the nutritional value of the fish product and improving the edible safety.
Drawings
FIG. 1 is a strain NJ molecular phylogenetic tree constructed based on 16S rRNA sequences;
FIG. 2 shows the effect of the ethanol extract of Ensifera on the growth curve of 2 strains of bacteria
FIG. 3 shows the effect of the extract of Ensifera on the conductivity of 2 strains of bacteria
FIG. 4 shows the effect of the extract of Ensifera indica (L.) Gaertn on the nucleic acid content of the bacterial suspension
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1 use of an extract of emsia crenata sims tea of the present invention in inhibiting putrefaction of fish meat cooked at high temperature,
the following examples used various raw materials and equipment sources and detection methods as follows:
1 materials and methods
1.1 materials and apparatus
DFY-1000 Chinese herbal medicine pulverizer (Wenling Lin big machinery Co., Ltd.), imark enzyme labeling instrument (Bio-Rad Burley), RE-200A rotary evaporator (Guangzhou Cix instruments Co., Ltd.), hermo Fisher Shier conductivity meter (model: 3-star), CR-80CO2 incubator (Guangzhou Kangheng instruments Co., Ltd.), etc.; the other reagents are all domestic analytical purifiers. 3 live carps (0.52kg, 0.63kg and 1.1kg) were purchased from the good and many supermarkets of Enshi City, vine tea samples were collected from the peripheral areas of Enshi City, and the high-level agronomers of the institute of Chinese medicinal materials of the academy of agricultural sciences of Hubei province were identified by the x articles.
L B culture medium is prepared from yeast extract 5 g/L, tryptone 10 g/L10 g/L, pH7.2-7.4, and sterilized at 121 deg.C for 30min (agar 20 g/L is added to solid L B culture medium).
MH liquid culture medium is prepared from beef powder 2 g/L, soluble starch 1.5 g/L, acid hydrolyzed casein 17.5 g/L, pH7.2-7.6, and sterilizing at 121 deg.C for 30 min.
1.2 test methods
1.2.1 ethanol extract of Enshi Ampelopsis grossedentata
Crushing 10g of dried vine tea, soaking the vine tea in ethanol with the material-liquid ratio of 1:50 and 250m L60% for 18h, ultrasonically extracting the vine tea for 30min with the power of 200w, centrifuging the vine tea at 10000rpm, taking supernatant, concentrating the vine tea ethanol extract into extract-shaped solid under the conditions of the pressure of-0.090 MPa and the temperature of 40 ℃, and obtaining 4.025g of vine tea ethanol extract without ethanol solvent, wherein the pressure of the vine tea ethanol extract is 0.090MPa, and the concentration of the vine tea ethanol extract is no drop by a rotary evaporator condenser.
1.2.2 preservation test of Enshi vine tea ethanol extract on high-temperature cooked Fish meat
Antiseptic test comprises cleaning fresh live fish, boiling 100g fish meat at 100 deg.C for 10min, mashing in a clean bench with sterile mortar, adding 100m L sterile water, mixing, and adding 5.0mg m L-1、10.0mg·mL-1、20.0mg·mL-1、30.0mg·mL-1、40.0mg·mL-1、50.0mg·mL-1The ethanol extract of (1) was used as experimental group, sterile water was used as blank control, ampicillin was added as positive control to a final concentration of 100. mu.g/m L, 3 replicates were set for each experimental group, 1000. mu.l of sample was applied evenly to L B solid plates and cultured at 37 ℃ with counting colonies every 24 h.
1.2.3 separation and identification of putrefying bacteria of fish cooked at high temperature
Taking 1000 ul of mashed fish tissue in 1.2.2, uniformly coating L B solid plate, culturing for 24h at 37 ℃, selecting colonies, streaking, purifying, amplifying, and quickly extracting colony genome DNA by a magnetic bead method, using 16S rRNA gene universal primers (F: 5' -AGAGAGTTTGATCCTGGCTCAG, R: GCTTACCTTGTTACGACTT), amplifying by Fastpfu high fidelity polymerase PCR, using dNTP 2 mu L.5 mu L, upstream and downstream primers each 1.25 mu L, colony genome DNA template 1 mu L, Fastpfu high fidelity polymerase 1 mu L, Fastpfu buffer 10 mu L, ddH 25 mu L, PCR conditions of pre-denaturation 4min at 94 ℃, denaturation 1min at 94 ℃, annealing 1min at 57 ℃, extension 1.5min at 72 ℃, 30 cycles, extension 10min at 72 ℃, heat preservation at 4 ℃, purifying kit (Axygen) purifying PCR products by gel recovery kit (Axygene), cloning on PCR products to 57 ℃, cloning to clone 1.5min at 72 ℃, using a gene library of transgenic strain library of transgenic horse radish tree (Johndse. origin), and identifying by using a genetic transformation system of DNA, PCR (gene library of transgenic strain).
1.2.4 Enshi Ampelopsis grossedentata ethanol extract antibacterial Activity test
Testing the bacteriostatic activity of the ethanol extract on 2 putrefying bacteria by using a filter paper method, and culturing to OD 20 mu L600Mixing the test bacteria liquid of 0.6 and solid culture medium of 20m L L B, pouring into a flat plate, collecting 15 μ L Ampelopsis grossedentata ethanol extract (dissolved in dimethyl sulfoxide DMSO), dripping onto filter paper (diameter 6mm) for three times, culturing the filter paper on the culture medium mixed with bacteria liquid, setting ampicillin (100 μ g/m L) as positive control and DMSO as blank control, culturing at 37 deg.C for 16h, measuring inhibition zone, and determining antibiotic susceptibility standard, high sensitivity (diameter of inhibition zone)>20mm), medium sensitivity (10mm-20mm), light sensitivity or drug resistance (<10mm) the minimum inhibitory concentration against spoilage bacteria of the ethanol extract was determined by broth microdilution according to the american society for clinical and laboratory standardization (C L SI) M07-a9 standard protocol by adding MH broth 100 μ L and ethanol extract samples to each well in 1-12 columns on a 96-well plate to give final concentrations of 128.0, 64.0, 32.0, 16.0, 8.0, 4.0, 2.0, 1.0, 0.5 and 0.25mg/M L (3-12 columns), respectively.Diluting the cultured bacterial liquid by 1000 times, adding 100 mu L of the diluted bacterial liquid into each well, setting ampicillin (100 mu g/M L) as a positive control, arranging the No. 1 as a blank culture medium control, arranging the No. 2 as a blank control, arranging the No. 3-12 as a sample test column, detecting OD600 by using an imark enzyme labeling instrument after culturing for 16h at 37 ℃, and comparing the concentration of the drug in a bacterial growth tube which inhibits 80% with that in a positive growth control tube according to C L SI M100-S26 to obtain the MIC of the tested bacteria.
1.2.5 antibacterial mechanism research of Enshi vine tea ethanol extract
Measuring the influence of ethanol extract on the growth of 2 putrefying bacteria by using an imark microplate reader, setting MH liquid culture medium 50m L containing ethanol extract with final concentration of 1 × MIC and 1/2 × MIC, setting MH liquid culture medium containing DMSO as blank control, performing shake culture at 37 ℃ and 150r/min for 24h, sampling every 2h, and detecting OD600Measuring the influence of the ethanol extract on the conductivity of 2 putrefying bacteria liquid by using a conductivity meter, washing the bacteria liquid in the logarithmic growth phase for 3 times by using 0.1 mol/L phosphate buffer solution (pH7.4), adjusting the concentration of the bacteria liquid to 108CFU/m L, respectively adding the ethanol extract with the final concentration of 1 × MIC, measuring the conductivity once every 10min, measuring the release amount of the ethanol extract on the dissolved matters in the 2 putrefying bacteria liquid by using an imark enzyme-labeling instrument, centrifuging the bacteria liquid in the logarithmic growth phase for 15min at 4000r/min, collecting the bacteria, washing the bacteria by using 0.1 mol/L phosphate buffer solution (pH7.4) for 3 times, adjusting the concentration of the bacteria liquid to 108CFU/m L by using 0.1 mol/L phosphate buffer solution (pH7.4), respectively adding the ethanol extracts with the final concentrations of 1 × and MIC 1/2 × MIC, culturing the bacteria liquid for 6h at 37 ℃, shaking table for 6h, sampling every 2h and 5min at 8000rpm, and measuring the OD260。
2 results and analysis
2.1 preservation test of Enshi Ampelopsis grossedentata ethanol extract on high-temperature cooked Fish meat
The change of the bacterial content in the storage process of the fish products is an important index for evaluating the quality of the fish products in the storage and fresh-keeping processes, and the table 1 shows that the vine tea ethanol extract treatment concentration is increased, the colony number growing in the fish cooked at high temperature is obviously reduced, and when the fish products are stored for 6 days, the treatment group is 5.0 mg.m L-1、10.0mg·mL-1、20.0mg·mL-1、30.0mg·mL-1And the colony counts of the blank control group (CK) were 263, 22 + -3, 11 + -2, 1 and 35 + -3, treatment group 10.0mg m L-1、20.0mg·mL-1、30.0mg·mL-1Achieves a very significant difference level (P) with CK<0.01) it can be thus obtained that the Ampelopsis grossedentata ethanol extract can inhibit the growth of bacteria in the fish meat cooked at high temperature, and 30.0 mg. m L in storage 15 days-1The colony number of the treated group is less than or equal to 1, and the final concentration is 30 mg.m L-1The above ampelopsis grossedentata ethanol extract can completely inhibit the growth of bacteria in the fish meat boiled at high temperature, as shown in Table 1.
Table 1 effect of ethanol extract treatment of various concentrations of emsia crenata on the number of colonies growing during storage of high-temperature cooked fish meat (means ± SD, n ═ 3)
Injecting, a) adding sterile water for treatment as a blank control;
b) adding 100 mu g m L as the final concentration-1Ampicillin as a positive control;
c) "-" indicates the formation of a lawn and failure to count.
2.2 separation and identification of putrefying bacteria in fish meat
During the cooking process of the fish, high temperature basically kills microbial thalli and spores which can not form spores, and the spores with strong stress resistance are the main reason for causing the fish to decay. 2 strains of Bacillus methylotrophicus T-10(Bacillus methylotrophicus T-10) and Alcaligenes faecalis (Alcaligenes faecalis T-9) isolated from high temperature cooked fish meat, and the RDPRelease 11 database shows that the strain Bacillus methylotrophicus T-10 belongs to Bacillus genus of Bacillus family 1, and shows 97.6% homology with the standard strain Bacillus subtilis DSM 10. The RDPRelease 11 database showed that the strain Alcaligenes faecalis T-9 belongs to the genus Alcaligenes of the Alcaligenes family, showing a homology of.94.6% with the standard strain Alcaligenes faecalis G. The phylogenetic tree is shown in figure 1, and a strain NJ molecular phylogenetic tree is constructed based on a 16S rRNA sequence.
2.3 Enshi Ampelopsis grossedentata ethanol extract antibacterial Activity test
As shown in Table 2, the pair of Ampelopsis grossedentata ethanol extracts2 fish spoilage bacteria have good bacteriostatic effect, and have moderate sensitive inhibitory activity (inhibition zone is 11-15mm) on Bacillus methylotrophicus T-10 and Alcaligenes faecalis T-9, and MIC values of the bacteria are respectively 18.5 mg.m L-1、19.2mg·mL-1. The bacillus and the alcaligenes faecalis can form spores at high temperature, are main putrefying bacteria causing high-temperature cooked meat products such as fishes and the like, and the Ensifera ampelopsis ethanol extract can effectively inhibit the germination of the spores.
TABLE 2 antibacterial Activity of Enshi Ampelopsis grossedentata ethanol extract (means + -SD, n ═ 3)
Note that a) 100. mu.g.m L final concentration was added-1Ampicillin as a positive control
2.4 bacteriostatic mechanism of the extract of Enshi vine tea
Comparing the blank control group (DMSO), the ampelopsis grossedentata ethanol extract treatment group with 1/2 × MIC concentration and the ampelopsis grossedentata ethanol extract treatment group with 1 × MIC concentration, after logarithmic phase and stationary phase delay, as shown in figure 2, the ampelopsis grossedentata ethanol extract has the effect of obviously inhibiting the multiplication of bacteria in logarithmic phase on 2 putrefying bacteria, when the ampelopsis grossedentata ethanol extract treatment concentration is increased from 1/2 × MIC to MIC, the growth inhibition effect on Bacillus methylotrophicus T-10 and Alcaligenes faecalis T-9 is increased, and the logarithmic phase and stationary phase further delay, and the thallus density is reduced.
FIG. 2 Effect of Enshi Ampelopsis grossedentata ethanol extract on growth curves of 2 strains
As shown in FIG. 3, comparing the sterile blank control group (adding Ampelopsis grossedentata ethanol extract with MIC concentration) and the bacterial liquid blank control group (DMSO), the conductivity of the Ampelopsis grossedentata ethanol extract treatment group with 1 × MIC concentration is decreased rapidly and increased, it is presumed that the Ensifera grossedentata ethanol extract causes the ion concentration of the bacterial liquid of Bacillus and Alcaligenes faecalis to be decreased rapidly and increased, it is presumed that the possible mechanism is that the organic acid compound in the ethanol extract is bonded to the surface of the cell membrane first, so that the conductivity of the bacterial liquid is decreased, the organic phenolic acid radical ion is bonded to the surfaceAfter binding to the surface of the cell membrane, the compound electrostatically interacts with the cell membrane to cause membrane damage and K in the bacterial cell+、Na+、H+And the leakage of charged ions such as nucleic acid, etc. and the increase of the conductivity of the bacterial liquid, thereby inhibiting the growth of bacteria.
FIG. 3 Effect of Ensify ampelopsis grossedentata ethanol extract on the conductivity of 2 strains of fungal fluid
As shown in FIG. 4, OD in the vine tea ethanol extract-treated group at 1/2 × MIC concentration and the vine tea ethanol extract-treated group at 1 × MIC concentration were compared in the blank control group (DMSO)260The method is characterized in that the concentration of nucleic acid in bacillus and alcaligenes faecalis is presumed to be increased by the ampelopsis grossedentata ethanol extract, and when the treatment concentration of the ethanol extract is increased from 1/2 × MIC to MIC, the concentration of the nucleic acid in the bacterial liquid is further increased.
FIG. 4 the effect of the ethanol extract of Ensifera on the nucleic acid content of the bacterial suspension
Discussion of 3
The natural preservative applied to fish preservation at present mainly comprises lactic acid bacteria, tea polyphenol and chitosan film. The lactobacillus biological preservative developed by utilizing the growth competitive power of the lactobacillus has good preservation effect on the yellow croaker flesh, the sea bass blocks and the tilapia; the tea polyphenols have antioxidant effect, and can effectively slow down chain reaction and fish protein oxidation in the fat oxidation process of the chub meat blocks, and prolong the preservation time of the chub blocks; the edible film of chitosan can inhibit the growth of mesophilic and psychrophilic aerobic putrefying bacteria of Lateolabrax japonicus meat, and prolong the shelf life of Lateolabrax japonicus meat by about 20 days. The lactobacillus is extremely susceptible to the environment as a microorganism, and the metabolite of the lactobacillus can influence the flavor and the quality of fish meat, so that the application of the lactobacillus in the preservation of aquatic products is limited. The extraction process of the tea polyphenol and the chitosan-based membrane has higher requirements, and the large-scale application of the chitosan-based membrane in fish preservation is limited.
The ampelopsis grossedentata ethanol extract contains various effective active ingredients: flavones, phenols, polysaccharides, volatile oil, amino acids, trace elements and the like, wherein the content of the flavones is 43.40-45.52%, and the main effective component dihydromyricetin in the flavones compound has the bacteriostatic action on staphylococcus aureus, multi-resistant pseudomonas aeruginosa, escherichia coli and bacillus subtilis and also has the antioxidant function. It is presumed that flavonoids such as dihydromyricetin in the ampelopsis grossedentata ethanol extract inhibits the reproduction of bacillus and alcaligenes faecalis in the high-temperature cooked fish meat, and can effectively slow down the chain reaction in the oxidation process of the fat in the fish meat blocks and the oxidation of the fish protein. The ampelopsis grossedentata ethanol extract has a simple extraction process, contains various active ingredients, is homologous in medicine and food, is green and healthy, can be used as a pure natural food preservative, and provides an important reference for prolonging the storage time of a fish product, improving the nutritional value of the fish product and researching the edible safety.
Example 2 application of an enrofloxacin vine tea ethanol extract in inhibiting high-temperature cooked fish decay, the application of the enrofloxacin vine tea ethanol extract in inhibiting high-temperature cooked fish decay, crushing dried vine tea, soaking for 17h in 50% ethanol according to a material-liquid ratio, carrying out ultrasonic leaching at a power of 190w for 28min, and carrying out vacuum concentration under the conditions that the pressure is-0.085 MPa and the temperature is 35 ℃, concentrating the vine tea ethanol extract into a pasty solid, and carrying out no-drop evaporation in a condensation pipe of a rotary evaporator to obtain a vine tea ethanol extract preservative solution without an ethanol solvent, wherein the vine tea ethanol extract is dissolved in sterile water to ensure that the concentration of the vine tea ethanol extract is 40.0 mg.m L-1Soaking cooked fish meat in water at a concentration of 40.0mg · m L-1Soaking in the preservative solution for 30 min.
Example 3 application of the enrofloxacin ethanol extract in inhibiting high-temperature cooked fish decay, the application of the enrofloxacin ethanol extract in inhibiting high-temperature cooked fish decay, crushing dried ampelopsis grossedentata, soaking for 19h according to a material-liquid ratio of 1:55 and 70% ethanol, carrying out ultrasonic leaching at a power of 210w for 32min, and carrying out concentration under reduced pressure at a pressure of-0.095 MPa and a temperature of 40 ℃, so that the ampelopsis grossedentata ethanol extract is concentrated into a pasty solid, and a rotary evaporator condensation tube does not have liquid drops, so that an ampelopsis grossedentata ethanol extract preservative solution without an ethanol solvent is obtained-1Soaking cooked fish meat in the solution with concentration of 50.0mg · m L-1Soaking in the preservative solution for 35 min.
Example 4 application of an enrofloxacin ethanol extract in inhibiting high-temperature cooked fish decay, the application of the enrofloxacin ethanol extract in inhibiting high-temperature cooked fish decay, crushing dried ampelopsis grossedentata, soaking for 18h according to a material-liquid ratio of 1:45 and 60% ethanol, ultrasonically leaching at a power of 200w for 30min, and concentrating under reduced pressure at a pressure of-0.090 MPa and a temperature of 38 ℃, concentrating the ampelopsis grossedentata ethanol extract into a pasty solid, and dissolving the ampelopsis grossedentata ethanol extract into sterile water to ensure that the concentration of the ampelopsis grossedentata ethanol extract is 45 mg.m L and no liquid drops are generated in a condensation pipe of a rotary evaporator to obtain an ethanol solvent-free ampelopsis grossedentata ethanol extract preservative solution-1Soaking cooked fish meat in water at a concentration of 45 mg. m L-1Soaking in the preservative solution for 32 min.
Example 5 application of an Ensifeng Ampelopsis grossedentata ethanol extract in inhibiting high-temperature cooked fish decay, and application of the Ensifeng Ampelopsis grossedentata ethanol extract in inhibiting high-temperature cooked fish decay, the Ensifeng Ampelopsis grossedentata ethanol extract is obtained by crushing 10g of dried Ampelopsis grossedentata, soaking the crushed dried Ampelopsis grossedentata in ethanol with the material-liquid ratio of 1:50 and 250m L60% for 18h, carrying out ultrasonic extraction with the power of 200w for 30min, centrifuging at 10000rpm, taking supernatant, concentrating under reduced pressure under the conditions that the pressure is-0.090 MPa and the temperature is 40 ℃, concentrating the Ampelopsis grossedentata ethanol extract into pasty solid, and dissolving the Ampelopsis grossedentata ethanol extract in sterile water to make the concentration of the ethanol extract be 40.0 mg.m L-1-50.0mg·mL-1Soaking cooked fish meat in water at a concentration of 40.0mg · m L-1-50.0mg·mL-1Soaking in the preservative solution for 30-35 min.
Example 6 application of the ethanol extract of embelia continentalis Bunge of the invention in inhibiting the putrefaction of fish cooked at high temperature, and application of the ethanol extract of embelia continentalis Bunge in inhibiting the putrefaction of fish cooked at high temperature, the ethanol extract of embelia continentalis Bunge is obtained by crushing 10g of dried ampelopsis grossedentata, soaking in ethanol with a concentration ratio of 1:50 to 250m L60% for 18h at a power of 200w and ultrasonic extraction for 30min, centrifuging at 10000rpm to obtain supernatant, concentrating under reduced pressure at a pressure of-0.090 MPa and a temperature of 40 ℃, concentrating the ethanol extract of embelia continentalis Bunge into pasty solid,the rotary evaporator condenser tube has no liquid drop to obtain Ampelopsis Grossdentata ethanol extract without ethanol solvent 4.025g, and the Ampelopsis Grossdentata ethanol extract is dissolved with sterile water to reach concentration of 50.0 mg.m L-1Soaking cooked fish meat in the solution with concentration of 50.0mg · m L-1Soaking in the preservative solution for 35 min.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (4)
1. The application of the Ensifeng tea ethanol extract in inhibiting the putrefaction of high-temperature cooked fish meat is characterized in that: the application of the Ensifera indica ethanol extract in inhibiting putrefaction of high-temperature cooked fish meat.
2. The use of an extract of emschynia indica ethyl alcohol according to claim 1 for inhibiting putrefaction of high temperature cooked fish meat, wherein: pulverizing dried Ampelopsis grossedentata, and soaking in 50% -70% ethanol at a material-liquid ratio of 1:40-55 for 17-19 h. Ultrasonic leaching at power of 190-210 w for 28-32 min, and concentrating under reduced pressure: concentrating the vine tea ethanol extract into pasty solid under the pressure of-0.085 MPa to-0.095 MPa and at the temperature of 35 ℃ to 40 ℃, and obtaining the vine tea ethanol extract preservative solution without ethanol solvent, wherein no liquid drop exists in a condensation tube of a rotary evaporator.
3. The use of the Ensify ampelopsis grossedentata ethanol extract for inhibiting putrefaction of fish meat cooked at high temperature according to claim 1, wherein the ampelopsis grossedentata ethanol extract is dissolved in sterile water to a concentration of 40.0 mg-m L-1-50.0mg·mL-1Soaking cooked fish meat in water at a concentration of 40.0mg · m L-1-50.0mg·mL-1Soaking in the preservative solution for 30-35 min.
4. The use of an extract of emschynia indica tea ethanol as claimed in claim 3 for inhibiting putrefaction of fish cooked at high temperature, characterized in thatThe ampelopsis grossedentata ethanol extract is dissolved by sterile water to ensure that the concentration of the ampelopsis grossedentata ethanol extract is 45 mg.m L-1Soaking cooked fish meat in water at a concentration of 45.0mg · m L-1Soaking in the antiseptic solution for 33 min.
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| CN111955636A (en) * | 2020-09-09 | 2020-11-20 | 湖北省农业科学院中药材研究所 | Composite vine tea solid beverage with antibacterial function |
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| CN105707202A (en) * | 2016-01-29 | 2016-06-29 | 广东省农业科学院蚕业与农产品加工研究所 | Natural plant extract dried fish preservative and preparation method and application thereof |
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| CN105707202A (en) * | 2016-01-29 | 2016-06-29 | 广东省农业科学院蚕业与农产品加工研究所 | Natural plant extract dried fish preservative and preparation method and application thereof |
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| 周茜雅等: "藤茶抗菌作用研究进展", 《中草药》 * |
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| CN111955636A (en) * | 2020-09-09 | 2020-11-20 | 湖北省农业科学院中药材研究所 | Composite vine tea solid beverage with antibacterial function |
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