CN110839687A - Method for preserving globefish by combining bacteria-reducing pretreatment with vacuum packaging - Google Patents
Method for preserving globefish by combining bacteria-reducing pretreatment with vacuum packaging Download PDFInfo
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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
- A23B4/015—Preserving by irradiation or electric treatment without heating effect
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- A—HUMAN NECESSITIES
- A22—BUTCHERING; MEAT TREATMENT; PROCESSING POULTRY OR FISH
- A22C—PROCESSING MEAT, POULTRY, OR FISH
- A22C25/00—Processing fish ; Curing of fish; Stunning of fish by electric current; Investigating fish by optical means
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- A—HUMAN NECESSITIES
- A22—BUTCHERING; MEAT TREATMENT; PROCESSING POULTRY OR FISH
- A22C—PROCESSING MEAT, POULTRY, OR FISH
- A22C25/00—Processing fish ; Curing of fish; Stunning of fish by electric current; Investigating fish by optical means
- A22C25/02—Washing or descaling fish
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- A—HUMAN NECESSITIES
- A22—BUTCHERING; MEAT TREATMENT; PROCESSING POULTRY OR FISH
- A22C—PROCESSING MEAT, POULTRY, OR FISH
- A22C25/00—Processing fish ; Curing of fish; Stunning of fish by electric current; Investigating fish by optical means
- A22C25/14—Beheading, eviscerating, or cleaning fish
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- A—HUMAN NECESSITIES
- A22—BUTCHERING; MEAT TREATMENT; PROCESSING POULTRY OR FISH
- A22C—PROCESSING MEAT, POULTRY, OR FISH
- A22C25/00—Processing fish ; Curing of fish; Stunning of fish by electric current; Investigating fish by optical means
- A22C25/14—Beheading, eviscerating, or cleaning fish
- A22C25/145—Eviscerating fish
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- A—HUMAN NECESSITIES
- A22—BUTCHERING; MEAT TREATMENT; PROCESSING POULTRY OR FISH
- A22C—PROCESSING MEAT, POULTRY, OR FISH
- A22C25/00—Processing fish ; Curing of fish; Stunning of fish by electric current; Investigating fish by optical means
- A22C25/17—Skinning fish
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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
- A23B4/06—Freezing; Subsequent thawing; Cooling
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Abstract
A method for preserving puffer fish by combining sterilization pretreatment with vacuum packaging comprises selecting fish sample, preparing slightly acidic electrolyzed water, processing fish sample, sterilizing, packaging into bags, vacuum packaging, and refrigerating with refrigerator. The invention can reduce the microbial quantity of the sample after the puffer fish is subjected to the sterilization treatment by the prepared WAEW, so that the total microbial quantity of the treated group sample in the storage process is in a relatively low level, meanwhile, the sample is isolated from oxygen by vacuum packaging, the growth and the propagation of aerobic microorganisms are further inhibited, the quality deterioration reactions of the puffer fish, such as protein degradation, fat oxidation and the like, are effectively delayed by the synergistic effect of the two, the shelf life of the puffer fish is prolonged, and meanwhile, the composite technology is safe and environment-friendly, has relatively low production cost, and is a puffer fish refrigeration and fresh-keeping means with high commercial value.
Description
Technical Field
The invention relates to a method for preserving puffer fish, in particular to a method for preserving puffer fish by combining sterilization pretreatment with vacuum packaging.
Background
The globefish is a traditional food in coastal areas of China, is widely distributed in the east sea, the yellow sea, the Bohai sea and the Yangtze river basin, and is one fish species of three delicacies in the Yangtze river. The puffer fish is delicious in taste and rich in nutrition, contains 8 essential amino acids and 10 nonessential amino acids, has a ratio of the essential amino acids to the nonessential amino acids (WTEAA) of 68.34 percent, is higher than an ideal mode of FAO/WHO, is a high-quality protein source, contains high-content mineral elements such as potassium, calcium, zinc and the like, has relatively low fat content, and is popular with people. However, fresh puffer fish is extremely easy to decay under the action of exogenous microorganisms and endogenous enzymes, so that the commercial value is rapidly reduced, and the exploration of an effective fresh-keeping mode has important significance for the puffer fish industry. The prior preservation technology commonly used for aquatic products comprises low-temperature storage, sterilization treatment, vacuum packaging, modified atmosphere packaging, biological preservative and the like, wherein subacid Electrolyzed Water (WAEW) in the sterilization pretreatment technology is a commonly used bactericide, and hypochlorous acid molecules serving as active ingredients can not only destroy cell walls of various bacteria, but also generate oxidation with intracellular protein or destroy the activity of phosphate dehydrogenase of the intracellular protein, so that sugar metabolism is disordered to kill cells, and the method has the advantages of good storage stability, mild action conditions and the like. The Vacuum Packaging preservation (VP) technology forms a low-oxygen storage environment by removing air in product packages, can effectively inhibit the growth and propagation of microorganisms and product oxidation, thereby prolonging the shelf life of products.
The invention with the patent application number of 201810855767.0 discloses a tilapia fillet preservation method (application date: 2018.07.31). This tilapia fillet is frozen by adding a color fixative into WAEW to prepare fresh-keeping ice, the invention can effectively inhibit the reproduction of microorganisms and the oxidation of myoglobin, and keep the original color of the fillet, but the friction action of crushed ice easily causes the mechanical damage of the fillet, thereby accelerating the damage of muscle tissues, and meanwhile, the single WAEW treatment cannot meet the requirement of consumers on the shelf life of the tilapia.
The invention patent with the patent application number of 201210086607.7 discloses a method for preserving single-frozen shrimp meat by using electrolyzed water (application date: 2012.03.29), WAEW is prepared by electrolyzing a sodium chloride solution, the safety problem of microorganisms in a soaked shrimp meat sample is effectively controlled, and the quality safety of the product is ensured, but hydrogen which is difficult to dissolve in water and is generated in the process of electrolyzing the sodium chloride solution has great potential safety hazard, and meanwhile, chlorine generated in the electrolytic process is dissolved in water to form substances such as sodium hypochlorite and the like with oxidation effect, and although the substances have a certain sterilization effect, the putrefaction rate of the shrimp meat is easier to accelerate by contacting the oxidized substances for a long time.
The invention with patent application number 201610228406.4 discloses a WAEW in a method for preserving bonito by weak acid electrolyzed water (application date: 2016.04.13), which is prepared from deionized water, sodium chloride, ferric chloride, copper sulfate, amylose, glycerol, a conductive agent, an antioxidant and the like, can effectively kill harmful bacteria such as staphylococcus aureus and the like and inhibit the growth of putrefying microorganisms, does not generate flammable and explosive hydrogen, but has more types of added reagents, and cannot ensure that the residues have no influence on the health of a human body,
WAEW is an effective bactericide in the range of food additives, is traditionally obtained by electrolyzing sodium chloride solution, and potential safety hazards are caused by flammability and explosiveness of hydrogen generated in the electrolysis process, so dilute hydrochloric acid is selected as the electrolytic solution. Vacuum packaging is a widely applied and obvious fresh-keeping mode in the aquatic product industry, so that the invention combines two fresh-keeping modes to be applied to the fresh-keeping process of the puffer fish, can achieve the synergistic fresh-keeping effect and caters to the concept that consumers pursue green and healthy products.
Disclosure of Invention
The invention aims to provide a puffer fish fresh-keeping technology combining sterilization pretreatment with vacuum packaging, which is characterized in that WAEW sterilization treatment is carried out, then vacuum packaging is carried out, and the puffer fish is stored at 4 ℃, so that the technology can effectively inhibit the growth and the reproduction of microorganisms in a sample, maintain the volatile basic nitrogen (TVB-N), the freshness value (K), the Trimethylamine (TMA) and the thiobarbituric acid value (TBA) at relatively low levels, slow down the dissolving amount of myofibrillar protein and the reduction of hardness value, and remarkably prolong the shelf life of the puffer fish.
The invention relates to a method for preserving globefish by combining the bacteria-reducing pretreatment with the vacuum packaging, which comprises the steps of selecting and purchasing a fish sample, preparing slightly acidic electrolyzed water, processing the fish sample, reducing the bacteria, packaging into a bag, vacuum packaging and refrigerating in a refrigerator, and is characterized by comprising the following operation steps:
(1) selecting and purchasing a fish sample: selecting puffer fish with uniform size, bright color, fresh state and weight of 300 + -10 g, and transporting to laboratory via foam box;
(2) preparing subacid electrolyzed water: preparing a dilute hydrochloric acid solution with the Concentration of 2.70-3.30%, starting a subacid electrolyzed water experimental machine, setting the voltage range to be 5.20-5.30V, the current range to be 2.90-3.00A and the flow rate range to be 0.97-1.03 ml/min, opening a 'sample injection pump' and an 'electrolytic bath' switch, preparing WAEW with the parameters of pH value of 6.21-6.56, oxidation-reduction Potential (ORP) of 843.3-907.6 mv and Available Chlorine Content (ACC) of 27.21-38.16 ppm, preparing the WAEW for 15-25 min, and taking a sufficient amount of WAEW for standby by using a light-shielding container after the WAEW is stabilized;
(3) processing the fish sample: rapidly cutting open the abdomen of the puffer fish with sharp scissors to bleed blood; after the gills are cut off, the gills and internal organs are removed together, and then the residual internal organs are trimmed by scissors; tearing off the skin of the puffer along the upper abdominal part cutting line of the puffer fish; removing the eyes of the puffer fish with scissors; squeezing out residual blood in the vertebral column of the globefish; cleaning the globefish with ice water;
(4) and (3) carrying out sterilization treatment: putting 2-14 parts of slaughtered and cleaned globefish into a raw material with a mass ratio of 1: 3-1: 6, soaking in WAEW for 8-15 min for sterilization treatment;
(5) packaging into a bag: respectively filling the puffer fish subjected to the bacteria reduction treatment into high-resistance polyvinylidene chloride packaging bags with the specification of 28 cm multiplied by 28 cm;
(6) and (3) vacuum packaging: starting a bag type air-conditioning fresh-locking packaging machine, starting a vacuum pump, selecting a vacuum packaging mode, setting the vacuumizing time to be 15-20 s, and the heat seal temperature to be 130-170 ℃, carrying out vacuum packaging on two groups of samples of a vacuum packaging group (VP) and a subacid electrolyzed water combined vacuum packaging group (WAEW + VP), placing the bag mouth of a packaging bag containing the globefish at a gas replacement and heat seal position, clicking a vacuumizing button to vacuumize, and finishing the vacuumizing process after the bag mouth is subjected to heat seal;
(7) refrigerating by a refrigerator: the four groups of puffer fish samples are quickly transferred into a refrigerator with the temperature of 4.0 +/-0.1 ℃ for storage. The vacuum packaging bag selected by the invention is a high-resistance polyvinylidene chloride packaging bag, and all samples are stored at the temperature of 4 +/-0.1 ℃.
The preferable scheme is as follows: the slightly acidic electrolyzed water used was prepared from a dilute hydrochloric acid solution having a concentration of 3.06%.
The preferable scheme is as follows: the parameters of the set slightly acidic electrolyzed water experimental machine are as follows: voltage 5.28V, current 3.02A, flow rate range 1.00 ml/min.
The preferable scheme is as follows: the parameters of the prepared subacid electrolyzed water are as follows: the pH value is 6.39, the oxidation-reduction Potential (ORP) ORP is 878.3mv, and the Available Chlorine Content (ACC) is 37.65 ppm.
The preferable scheme is as follows: the fish sample processing method comprises the following steps: rapidly cutting open the abdomen of the puffer fish with sharp scissors to bleed blood; after the gills are cut off, the gills and internal organs are removed together, and then the residual internal organs are trimmed by scissors; tearing off the skin of the puffer along the upper abdominal part cutting line of the puffer fish; removing the eyes of the puffer fish with scissors; squeezing out residual blood in the vertebral column of the globefish; cleaning the globefish with ice water;
the preferable scheme is as follows: during the sterilization treatment, the puffer fish is put into a container with the mass ratio of 1: 4 in WAEW for 10 min.
The preferable scheme is as follows: the packaging specification of the sterilized globefish is 28 cm multiplied by 28 cm high-resistance polyvinylidene chloride packaging bag.
The preferable scheme is as follows: the used vacuum packaging machine is a bag type air-adjusting fresh-locking packaging machine, and the vacuum packaging parameters are as follows: the vacuumizing time is 15 s, and the heat sealing temperature is 150 ℃.
The invention can reduce the microbial quantity of the sample after the puffer fish is subjected to the sterilization treatment by the prepared WAEW, so that the total microbial quantity of the treated group sample in the storage process is in a relatively low level, meanwhile, the sample is isolated from oxygen by vacuum packaging, the growth and the propagation of aerobic microorganisms are further inhibited, the quality deterioration reactions of the puffer fish, such as protein degradation, fat oxidation and the like, are effectively delayed by the synergistic effect of the two, the shelf life of the puffer fish is prolonged, and meanwhile, the composite technology is safe and environment-friendly, has relatively low production cost, and is a puffer fish refrigeration and fresh-keeping means with high commercial value.
Drawings
FIG. 1 is a graph showing the effect of slightly acidic electrolyzed water of different pH values on the total number of colonies of fresh puffer fish.
FIG. 2 is a graph showing the change in the total number of colonies (TVC) of the puffer fish packed in combination with slightly acidic electrolyzed water and vacuum.
FIG. 3 is a graph of the variation of the volatile basic nitrogen content (TVB-N) of puffer fish in combination with slightly acidic electrolyzed water and vacuum packaging.
Fig. 4 is a graph showing changes in the freshness value (K value) of slightly acidic electrolyzed water combined with vacuum packed globefish.
FIG. 5 shows the change of the Thiobabarbituric acid number (TBA value) of puffer fish in combination with slightly acidic electrolyzed water and vacuum packaging.
FIG. 6 shows the Trimethylamine (TMA) change in slightly acidic electrolyzed water in combination with vacuum packed puffer fish.
Figure 7 is a graph of the change in the amount of slightly acidic electrolyzed water combined with the amount of myofibrillar proteins from a vacuum packed puffer fish.
Figure 8 is a graph of the change in hardness of puffer fish in combination with slightly acidic electrolyzed water and vacuum packaging.
Detailed Description
To explain the operation flow and creation features of the present invention in more detail for the user to understand and use better, the following detailed description is given with reference to the embodiments.
Example 1: a method for preserving puffer fish by combining the sterilization pretreatment with vacuum packaging comprises the following steps:
(1) selecting and purchasing a fish sample: selecting puffer fish with uniform size, bright color, fresh state and weight of 300 + -10 g, and transporting to laboratory via foam box;
(2) preparing subacid electrolyzed water: preparing a dilute hydrochloric acid solution with the Concentration of 2.78%, starting a subacid electrolyzed water tester, setting the voltage to be 5.25V, the current to be 2.96A, the flow rate range to be 0.98 ml/min, opening a 'sample injection pump' and an 'electrolysis bath' switch, preparing WAEW with the parameters of pH value of 6.21, oxidation-reduction Potential (ORP) ORP of 852.4mv and Available Chlorine Content (ACC) of 28.33 ppm, preparing the WAEW for 18 min, and taking a sufficient amount of WAEW for standby by using a light-proof container after the WAEW is stabilized;
(3) processing the fish sample: rapidly cutting open the abdomen of the puffer fish with sharp scissors to bleed blood; after the gills are cut off, the gills and internal organs are removed together, and then the residual internal organs are trimmed by scissors; tearing off the skin of the puffer along the upper abdominal part cutting line of the puffer fish; removing the eyes of the puffer fish with scissors; squeezing out residual blood in the vertebral column of the globefish; cleaning the globefish with ice water;
(4) and (3) carrying out sterilization treatment: putting 2 parts of slaughtered and cleaned globefish into a reaction kettle with a volume ratio of 1: 3, soaking in WAEW for 8 min for sterilization treatment;
(5) sampling and detecting: and (3) detecting the total number of colonies of the treated 2 globefish, firstly taking 5.00 g of fish sample, putting the fish sample into 45mL of sterilized normal saline, and uniformly mixing to prepare a mixture with the weight ratio of 1:10, a homogeneous dilution; then 1ml sterile pipetting is used for sucking 1: injecting 1ml of 10 diluent into a test tube containing 9 ml of sterile normal saline along the wall for 10-time gradient dilution to prepare 1:100 sample diluent, uniformly mixing, and performing 10-time serial dilution according to the operation and actual conditions; selecting 3 bacteria liquid with proper dilution concentration, sucking 1mL of the bacteria liquid into sterilized culture dishes, enabling each gradient to be parallel to 2, simultaneously sucking 1mL of sterilized normal saline into the two sterilized culture dishes respectively, and placing the sterilized normal saline in a super clean bench to be used as a blank control; pouring 15-20 mL of plate counting agar culture medium cooled to 45 ℃ into the culture dish, and rotating the culture dish to uniformly mix the agar culture medium and the plate counting agar culture medium; after the agar is solidified, the flat plate is placed in a biochemical incubator at the temperature of 30 +/-1 ℃ upside down for culturing for 72 +/-3 h, the total number of bacterial colonies is determined by adopting a flat plate counting method, and sterilized normal saline is used as a blank for a contrast test;
(6) plate counting: and selecting a plate with the total number of colonies of 30-300 for counting.
Example 2: a method for preserving puffer fish by combining the sterilization pretreatment with vacuum packaging comprises the following steps:
(1) selecting and purchasing a fish sample: selecting puffer fish with uniform size, bright color, fresh state and weight of 300 + -10 g, and transporting to laboratory via foam box;
(2) preparing subacid electrolyzed water: preparing a dilute hydrochloric acid solution with the Concentration of 2.92%, starting a subacid electrolyzed water tester, setting the voltage to be 5.27V, the current to be 2.98A and the flow rate range to be 0.99 ml/min, opening a 'sample injection pump' and an 'electrolysis bath' switch, preparing WAEW with the parameters of pH value of 6.32, oxidation-reduction Potential (ORP) ORP of 861.1mv and Available Chlorine Content (ACC) of 30.27 ppm, preparing the WAEW for 19 min, and taking a sufficient amount of WAEW for standby by using a light-proof container after the WAEW is stabilized;
(3) processing the fish sample: rapidly cutting open the abdomen of the puffer fish with sharp scissors to bleed blood; after the gills are cut off, the gills and internal organs are removed together, and then the residual internal organs are trimmed by scissors; tearing off the skin of the puffer along the upper abdominal part cutting line of the puffer fish; removing the eyes of the puffer fish with scissors; squeezing out residual blood in the vertebral column of the globefish; cleaning the globefish with ice water;
(4) and (3) carrying out sterilization treatment: putting 2 parts of slaughtered and cleaned globefish into a reaction kettle with a volume ratio of 1: 4, soaking in WAEW for 8 min for sterilization treatment;
(5) sampling and detecting: and (3) detecting the total number of colonies of the treated 2 puffer fish, namely, firstly taking 5.00 g of fish sample, uniformly mixing the fish sample in 45mL of sterilized normal saline, and preparing into a mixture with the weight ratio of 1:10, a homogeneous dilution; then 1ml sterile pipetting is used for sucking 1: injecting 1ml of 10 diluent into a test tube containing 9 ml of sterile normal saline along the wall for 10-time gradient dilution to prepare 1:100 sample diluent, uniformly mixing, and performing 10-time serial dilution according to the operation and actual conditions; selecting 3 bacteria liquid with proper dilution concentration, sucking 1mL of the bacteria liquid into sterilized culture dishes, enabling each gradient to be parallel to 2, simultaneously sucking 1mL of sterilized normal saline into the two sterilized culture dishes respectively, and placing the sterilized normal saline in a super clean bench to be used as a blank control; pouring 15-20 mL of plate counting agar culture medium cooled to 45 ℃ into the culture dish, and rotating the culture dish to uniformly mix the agar culture medium and the plate counting agar culture medium; after the agar is solidified, the flat plate is placed in a biochemical incubator at the temperature of 30 +/-1 ℃ upside down for culturing for 72 +/-3 h, the total number of bacterial colonies is determined by adopting a flat plate counting method, and sterilized normal saline is used as a blank for a contrast test;
(6) plate counting: and selecting a plate with the total number of colonies of 30-300 for counting.
Example 3: a method for preserving puffer fish by combining the sterilization pretreatment with vacuum packaging comprises the following steps:
(1) selecting and purchasing a fish sample: selecting puffer fish with uniform size, bright color, fresh state and weight of 300 + -10 g, and transporting to laboratory via foam box;
(2) preparing subacid electrolyzed water: preparing a dilute hydrochloric acid solution with the Concentration of 3.06%, starting a subacid electrolyzed water tester, setting the voltage to be 5.28V, the current to be 3.02A and the flow rate range to be 1.00 ml/min, opening a 'sample injection pump' and an 'electrolysis bath' switch, preparing WAEW with the parameters of pH value of 6.39, oxidation-reduction Potential (ORP) ORP of 878.3mv and Available Chlorine Content (ACC) of 37.65 ppm, preparing the WAEW for 20 min, and taking a sufficient amount of WAEW for standby after stabilization by using a light-shielding container;
(3) processing the fish sample: rapidly cutting open the abdomen of the puffer fish with sharp scissors to bleed blood; after the gills are cut off, the gills and internal organs are removed together, and then the residual internal organs are trimmed by scissors; tearing off the skin of the puffer along the upper abdominal part cutting line of the puffer fish; removing the eyes of the puffer fish with scissors; squeezing out residual blood in the vertebral column of the globefish; cleaning the globefish with ice water;
(4) and (3) carrying out sterilization treatment: putting 2 parts of slaughtered and cleaned globefish into a reaction kettle with a volume ratio of 1: 4, soaking in WAEW for 8 min for sterilization treatment;
(5) sampling and detecting: and (3) detecting the total number of colonies of the treated 2 puffer fish, namely, firstly taking 5.00 g of fish sample, uniformly mixing the fish sample in 45mL of sterilized normal saline, and preparing into a mixture with the weight ratio of 1:10, a homogeneous dilution; then 1ml sterile pipetting is used for sucking 1: injecting 1ml of 10 diluent into a test tube containing 9 ml of sterile normal saline along the wall for 10-time gradient dilution to prepare 1:100 sample diluent, uniformly mixing, and performing 10-time serial dilution according to the operation and actual conditions; selecting 3 bacteria liquid with proper dilution concentration, sucking 1mL of the bacteria liquid into sterilized culture dishes, enabling each gradient to be parallel to 2, simultaneously sucking 1mL of sterilized normal saline into the two sterilized culture dishes respectively, and placing the sterilized normal saline in a super clean bench to be used as a blank control; pouring 15-20 mL of plate counting agar culture medium cooled to 45 ℃ into the culture dish, and rotating the culture dish to uniformly mix the agar culture medium and the plate counting agar culture medium; after the agar is solidified, the flat plate is placed in a biochemical incubator at the temperature of 30 +/-1 ℃ upside down for culturing for 72 +/-3 h, the total number of bacterial colonies is determined by adopting a flat plate counting method, and sterilized normal saline is used as a blank for a contrast test;
(6) plate counting: and selecting a plate with the total number of colonies of 30-300 for counting.
Example 4
A method for preserving puffer fish by combining the sterilization pretreatment with vacuum packaging comprises the following steps:
(1) selecting and purchasing a fish sample: selecting puffer fish with uniform size, bright color, fresh state and weight of 300 + -10 g, and transporting to laboratory via foam box;
(2) processing the fish sample: rapidly cutting open the abdomen of the puffer fish with sharp scissors to bleed blood; after the gills are cut off, the gills and internal organs are removed together, and then the residual internal organs are trimmed by scissors; tearing off the skin of the puffer along the upper abdominal part cutting line of the puffer fish; removing the eyes of the puffer fish with scissors; squeezing out residual blood in the vertebral column of the globefish; cleaning the globefish with ice water;
(3) packaging into a bag: dividing the puffer fish into Air (AP) and vacuum packaging group (VP), and respectively packaging into high-resistance polyvinylidene chloride (PVDF) packaging bags of 28 cm × 28 cm;
(4) and (3) vacuum packaging: starting the bag type air-conditioning fresh-locking packaging machine, starting a vacuum pump, selecting a vacuum packaging mode, setting the vacuumizing time to be 15 s, and the heat sealing temperature to be 150 ℃, carrying out vacuum packaging on puffer fish in a vacuum packaging group (VP), placing the mouth of a vacuum packaging bag containing the puffer fish at a gas replacement and heat sealing position, clicking a vacuumizing button to vacuumize, and finishing the vacuumizing process after the mouth of the bag is heat sealed;
(5) refrigerating by a refrigerator: rapidly transferring the processed puffer fish sample into a refrigerator with the temperature of 4.0 +/-0.1 ℃ for storage;
(6) sampling and detecting: sampling detection is carried out for 0, 3, 6, 9, 12, 15 and 18 days respectively, the reserved 2 parts of puffer fish are detected for 0 day, after 0 day, two puffer fish are randomly sampled each time to be used as a parallel group for detection, and the influence of vacuum packaging on the fresh-keeping effect of the puffer fish is explored.
Example 5
A method for preserving puffer fish by combining the sterilization pretreatment with vacuum packaging comprises the following steps:
(1) selecting and purchasing a fish sample: selecting puffer fish with uniform size, bright color, fresh state and weight of 300 + -10 g, and transporting to laboratory via foam box;
(2) preparing subacid electrolyzed water: preparing a dilute hydrochloric acid solution with the Concentration of 3.06%, starting a subacid electrolyzed water tester, setting the voltage to be 5.28V, the current to be 3.02A and the flow rate range to be 1.00 ml/min, opening a 'sample injection pump' and an 'electrolysis bath' switch, preparing WAEW with the parameters of pH value of 6.39, oxidation-reduction Potential (ORP) ORP of 878.3mv and Available Chlorine Content (ACC) of 37.65 ppm, preparing the WAEW for 20 min, and taking a sufficient amount of WAEW for standby after stabilization by using a light-shielding container;
(3) processing the fish sample: rapidly cutting open the abdomen of the puffer fish with sharp scissors to bleed blood; after the gills are cut off, the gills and internal organs are removed together, and then the residual internal organs are trimmed by scissors; tearing off the skin of the puffer along the upper abdominal part cutting line of the puffer fish; removing the eyes of the puffer fish with scissors; squeezing out residual blood in the vertebral column of the globefish; cleaning the globefish with ice water;
(4) and (3) carrying out sterilization treatment: putting the slaughtered and cleaned globefish in 12 parts by volume ratio of 1: 4, soaking in WAEW for 10min for sterilization treatment;
(5) packaging into a bag: respectively filling 12 parts of puffer fish subjected to sterilization treatment into high-resistance polyvinylidene chloride packaging bags with the specification of 28 cm multiplied by 28 cm;
(6) and (3) vacuum packaging: starting the bag type air-conditioning fresh-keeping packaging machine, starting a vacuum pump, selecting a vacuum packaging mode, setting the vacuumizing time for 15 s, and the heat sealing temperature for 150 ℃, carrying out vacuum packaging on two groups of puffer fishes of a vacuum packaging group (VP) and a subacid electrolyzed water combined vacuum packaging group (WAEW + VP), placing the bag mouth of a vacuum packaging bag containing the puffer fishes at a gas replacement and heat sealing position, clicking a vacuumizing button to vacuumize, and finishing the vacuumizing process after the bag mouth is heat sealed;
(7) refrigerating by a refrigerator: rapidly transferring the four groups of processed puffer fish samples into a refrigerator with the temperature of 4.0 +/-0.1 ℃ for storage;
(8) sampling and detecting: sampling detection is carried out for 0, 3, 6, 9, 12, 15 and 18 days respectively, the reserved 2 parts of puffer fish are detected for 0 day, after 0 day, two puffer fish are randomly sampled each time and are used as a parallel group for detection, and the influence of slightly acidic electrolyzed water combined with vacuum packaging on the fresh-keeping effect of the puffer fish is researched.
The treated puffer fish is subjected to the following performance tests:
sampling and detecting: sampling and detecting for 0, 3, 6, 9, 12, 15 and 18 days respectively, detecting for 0 d for reserved 2 puffer fish, randomly sampling two puffer fish after 0 d each time to detect as parallel groups, and exploring the fresh-keeping effect of different groups of puffer fish through index measurement.
Index measurement:
(1) total number of colonies (TVC): accurately weighing 5.00 g of fish sample, putting into 45mL of sterilized normal saline, and uniformly mixing to obtain a mixture of 1:10, a homogeneous dilution; then 1ml sterile pipetting is used for sucking 1: injecting 1ml of 10 diluent into a test tube containing 9 ml of sterile normal saline along the wall for 10-time gradient dilution to prepare 1:100 sample diluent, uniformly mixing, and performing 10-time serial dilution according to the operation and actual conditions; selecting 3 bacteria liquid with proper dilution concentration, sucking 1mL of the bacteria liquid into sterilized culture dishes, enabling each gradient to be parallel to 2, simultaneously sucking 1mL of sterilized normal saline into the two sterilized culture dishes respectively, and placing the sterilized normal saline in a super clean bench to be used as a blank control; pouring 15-20 mL of plate counting agar culture medium cooled to 45 ℃ into the culture dish, and rotating the culture dish to uniformly mix the agar culture medium and the plate counting agar culture medium; after the agar is solidified, the flat plate is placed in a biochemical incubator at the temperature of 30 +/-1 ℃ upside down for culturing for 72 +/-3 h, the total number of bacterial colonies is determined by adopting a flat plate counting method, and sterilized normal saline is used as a blank for a contrast test; and after the culture, selecting a plate with the total number of colonies of 30-300 for counting.
(2) Volatile basic nitrogen content (TVB-N): the determination is carried out according to GB 5009.228-2016 (determination of volatile basic nitrogen in national food safety standard) and a semi-trace Kjeldahl nitrogen determination principle. Accurately weighing 5.00 g of puffer fish sample, placing in a digestive tube, adding 2 spoons of MgO powder, measuring the TVB-N value of the puffer fish sample by using a programmed Kjeldahl apparatus, performing 3 parallel experiments, and averaging.
(3) Freshness value (K value): accurately weighing 5.00 g of fish sample in a centrifugal tube, adding 10 mL of perchloric acid with volume fraction of 10% at 4 ℃, homogenizing, centrifuging for 15min at 4 ℃ and 8000r/min, and taking supernatant; adding 10 mL of perchloric acid with volume fraction of 5% at 4 ℃ into the precipitate, homogenizing, centrifuging at 4 ℃ and 8000r/min for 10min, and repeating for 2 times; finally, combining the supernatants, adding 15 mL of ultrapure water, adjusting the pH of the supernatant to 6.5 by using 10 mol/L and 1 mol/L KOH solutions and 5% and 10% perchloric acid solutions, standing for 30 min, taking the supernatant into a 50mL volumetric flask, metering the volume to 50mL by using the ultrapure water, and shaking up; finally, the mixture was filtered through a 0.22 μm membrane, and the filtrate was stored in a liquid phase sample bottle and subjected to high performance liquid analysis. 3 replicates of each sample were taken and averaged. High performance liquid phase analysis conditions: chromatography column VP-CDSC18 (46 mm. times.150 mm), which was eluted in equilibrium with 0.05 mol/L phosphate buffer pH =6.7, sample load of 10 uL, flow rate of 1mL/min, column temperature of 30 ℃, detection wavelength of 254 nm, and K value was calculated according to the formula, wherein WHxR, WHx, WATP, WADP, WAMP and WIMP are the mass fractions of inosine, hypoxanthine, adenosine triphosphate, adenosine diphosphate, adenylic acid and inosinic acid, respectively:
(4) thiobarbituric acid number (TBA): accurately weighing 5.00 g of fish sample homogenate, adding 25 mL of 20% trichloroacetic acid, standing for 1h, centrifuging at 0 deg.C and 8000r/min for 10min, filtering, transferring the filtrate to a 50mL volumetric flask, and fixing the volume. Transferring 5mL of the solution into a beaker, adding 5mL of 0.02 mol/L thiobarbituric acid solution, oscillating, mixing uniformly, reacting in a boiling water bath for 20 min, taking out and cooling to room temperature. The absorbance values were read at 532 nm against distilled water. The TBA values are as follows:. Wherein A is the absorbance value at 532 nm and TBA is in mg/kg.
(5) Trimethylamine (TMA): accurately weighing 2.00 g of fish sample, homogenizing, and centrifuging at 8000r/min for 10 min; placing 5ml of supernatant in a test tube, adding 1ml of formaldehyde (10%) solution, 10 ml of anhydrous toluene and 3 ml of 25% potassium hydroxide solution, repeatedly shaking, and heating at 30 deg.C for 5 min; discarding the water layer, and taking 7-9 ml toluene layer to contain 0.5 g anhydrous Na2SO4The resulting solution was dehydrated by shaking, 5mL of the toluene layer after the reaction was transferred to another tube containing 5mL of 0.02% picric acid solution, and the absorbance A1 was measured at 410 nm using a spectrophotometer. The blank was run with 7.5% TCA following the same procedure and the absorbance A0 was measured. Standard Curve by TMA (purity)>98%) and expressed as mg/100 g of sample, the absorbance is maintained at 30 ℃.
(6) Myofibrillar protein elution amount: firstly extracting myofibrillar protein, accurately weighing 2.00 g of fish meat, placing the fish meat into a 50mL centrifuge tube, adding 20 mL buffer solution A (20 mmol/L Tris-maleate, pH 7.0 and 0.05 mol/L KCl) for homogenization, freezing and centrifuging (4 ℃, 10000 Xg) for 15min, then discarding supernatant, repeatedly washing twice, adding 20 mL buffer solution B (20 mmol/L Tris-maleate, pH 7.0 and 0.6 mol/L KCl) into sediment for homogenization, then extracting for 1h at 4 ℃, freezing and centrifuging at 10000 r/min for 15min, wherein the supernatant is a myofibrillar protein solution, and measuring the concentration of the dissolved amount of the myofibrillar protein by adopting biuret. (7) Hardness value: cutting 15 mm × 15 mm × 15 mm fish blocks, selecting TPA mode to perform measurement, and setting parameters as follows: probe descent speed before measurement: 2.00 mm/s; testing speed: 1.00 mm/s; measuring the return speed of the probe: 5.00 mm/s; compression ratio: 40 percent; trigger force: 5.0 g; probe type p/5; the data acquisition rate was 200.00 points/s, and 3 parallel experiments were performed on each group of samples, and the average was taken.
The following table shows the indexes of the present invention and the control group after 18 days storage, wherein the present invention is the electrolyzed water treatment combined vacuum packaging group (WAEW + VP), and the control group is ① air packaging group (AP) ② vacuum packaging group (VP) ③ electrolyzed water treatment combined air packaging group (WAEW + AP):
| WAEW+VP | AP | VP | WAEW+AP | |
| TVC | 7.91 | 10.59 | 8.58 | 9.87 |
| TVB-N (mg/100g) | 22.23 | 47.11 | 24.26 | 36.61 |
| K value (%) | 59.50 | 75.21 | 63.99 | 71.79 |
| TBA (mg MDA/100g) | 0.63 | 0.75 | 0.70 | 0.73 |
| TMA (mg/100g) | 5.25 | 6.95 | 6.11 | 6.33 |
| amount of myofibrillar protein eluted (mg/g) | 31.30 | 25.15 | 29.38 | 26.50 |
| Hardness value (10)4) | 2.24 | 1.87 | 2.09 | 2.01 |
Through the comparison detection, all indexes of the puffer fish treated by the preservation method are superior to those of a control group, the shelf life of the puffer fish is obviously prolonged, and the preservation effect is good.
The experimental result of the invention shows that WAEW sterilization pretreatment is combined with vacuum packaging to effectively maintain the good storage quality of the puffer fish, the shelf life is prolonged from 7 days to 11 days of air packaging, and the technical method is simple to operate, safe and environment-friendly, and is an efficient and feasible puffer fish preservation technology.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.
Claims (7)
1. A method for preserving puffer fish by combining the sterilization pretreatment with vacuum packaging is characterized in that: the operation steps comprise fish sample selection, subacid electrolyzed water preparation, fish sample treatment, sterilization treatment, packaging and bagging, vacuum packaging and refrigerator refrigeration, and are characterized by comprising the following operation steps:
(1) selecting and purchasing a fish sample: selecting puffer fish with uniform size, bright color, fresh state and weight of 300 + -10 g, and transporting to laboratory via foam box;
(2) preparing subacid electrolyzed water: preparing a dilute hydrochloric acid solution with the concentration of 2.70-3.30%, starting a subacid electrolyzed water experimental machine, setting the voltage range to be 5.20-5.30V, the current range to be 2.90-3.00A and the flow rate range to be 0.97-1.03 ml/min, opening a 'sample feeding pump' and an 'electrolytic bath' switch, preparing WAEW with the parameters of pH value of 6.21-6.56, oxidation-reduction potential ORP of 843.3-907.6 mv and effective chlorine content ACC of 27.21-35.16 ppm, preparing the WAEW for 15-25 min, and taking a sufficient amount of WAEW for standby by using a light-shielding container after the WAEW is stable;
(3) processing the fish sample: rapidly cutting open the abdomen of the puffer fish with sharp scissors to bleed blood; after the gills are cut off, the gills and internal organs are removed together, and then the residual internal organs are trimmed by scissors; tearing off the skin of the puffer along the upper abdominal part cutting line of the puffer fish; removing the eyes of the puffer fish with scissors; squeezing out residual blood in the vertebral column of the globefish; cleaning the globefish with ice water;
(4) and (3) carrying out sterilization treatment: putting 2-14 parts of slaughtered and cleaned globefish into a raw material with a mass ratio of 1: 3-1: 6, soaking in WAEW for 8-15 min for sterilization treatment;
(5) packaging into a bag: respectively filling the puffer fish subjected to the bacteria reduction treatment into high-resistance polyvinylidene chloride packaging bags with the specification of 28 cm multiplied by 28 cm;
(6) and (3) vacuum packaging: starting a bag type air-conditioning fresh-locking packaging machine, starting a vacuum pump, selecting a vacuum packaging mode, setting the vacuumizing time to be 15-20 s, and the heat seal temperature to be 130-170 ℃, carrying out vacuum packaging on two groups of samples of a vacuum packaging group and a subacid electrolyzed water combined vacuum packaging group, placing the bag opening of a packaging bag containing puffer fish at a gas replacement and heat seal position, clicking a vacuumizing button to vacuumize, and finishing the vacuumizing process after the bag opening is heat sealed;
(7) refrigerating by a refrigerator: the four groups of puffer fish samples are quickly transferred into a refrigerator with the temperature of 4.0 +/-0.1 ℃ for storage.
2. The method for preserving puffer fish by combining the sterilization pretreatment with the vacuum packaging according to claim 1, wherein the method comprises the following steps:
the slightly acidic electrolyzed water used was prepared from a dilute hydrochloric acid solution having a concentration of 3.06%.
3. The method for preserving puffer fish by combining the sterilization pretreatment with the vacuum packaging according to claim 1, wherein the method comprises the following steps:
the parameters of the slightly acidic electrolyzed water experimental machine are as follows: voltage 5.28V, current 3.02A, flow rate range 1.00 ml/min.
4. The method for preserving puffer fish by combining the sterilization pretreatment with the vacuum packaging according to claim 1, wherein the method comprises the following steps:
the parameters of the prepared subacid electrolyzed water are as follows: the pH value is 6.39, the oxidation-reduction potential ORP is 878.3mv, and the available chlorine content ACC is 37.65 ppm.
5. The method for preserving puffer fish by combining the sterilization pretreatment with the vacuum packaging according to claim 1, wherein the method comprises the following steps:
during the sterilization treatment, the puffer fish is put into a container with the mass ratio of 1: 4 in WAEW for 10 min.
6. The method for preserving puffer fish by combining the sterilization pretreatment with the vacuum packaging according to claim 1, wherein the method comprises the following steps:
the packaging specification of the sterilized globefish is 28 cm multiplied by 28 cm high-resistance polyvinylidene chloride packaging bag.
7. The method for preserving puffer fish by combining the sterilization pretreatment with the vacuum packaging according to claim 1, wherein the method comprises the following steps:
the used vacuum packaging machine is a bag type air-adjusting fresh-locking packaging machine, and the vacuum packaging parameters are as follows: the vacuumizing time is 15 s, and the heat sealing temperature is 150 ℃.
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