CN104855489A - Processing method for decapterus maruadsi can - Google Patents

Abstract

A processing method for a decapterus maruadsi can relates to decapterus maruadsi. The invention provides the processing method for the decapterus maruadsi can, wherein the content of histamine of the decapterus maruadsi and the sensitivity of parvalbumin are reduced simultaneously, and the processing method is simple and reasonable in process, strong in operability, easy for scaled production and low in product sensitization. The processing method comprises the following steps of: abandoning decapterus maruadsi heads, tails and viscera and then cleaning the decapterus maruadsi; slicing fish meat to fish cubes, cleaning and then soaking the fish cubes in sweet water; putting the soaked fish meat into disinfected empty tanks; adding sweet water into the tanks and thermally evaporating; thermally sealing the tanks; sterilizing, cooling the tanks; and casing the tanks into boxes and putting the boxes into a storeroom. In the production and processing course of the decapterus maruadsi can, the sweet water is added, reducing sugars conduct Maillard reaction with histamine and parvalbumin of the fish meat during steps such as thermal evaporation, and meanwhile damaging the parvalbumin structure in the fish meat by virtue of high-temperature and high-pressure treatment; special texture and nutritional components of decapterus maruadsi are maintained to the maximum extent; at the same time, sensitive substance histamine and parvalbumin are reduced.

Description

A kind of processing method of indigo plant circle Scad can

Technical field

The present invention relates to blue circle Scad, especially relate to a kind of processing method of indigo plant circle Scad can.

Background technology

Consumer can cause some health problems when eating fish, and as parasitic disease, heavy metals exceeding standard, allergic reaction etc., wherein food hypersenstivity becomes the subject matter of global concern gradually.Research about fish anaphylaxis problem is divided into two classes, and a class is that allergic food is poisoning, as the histamine in the corruption flesh of fish; Two is hypersensitivity that anaphylactogen causes, as flesh of fish anaphylactogen parvalbumin.

Histamine (Histamine) be free histidine degrade under the effect of microorganism histidine decarboxylase produce a kind of amine, be extensively present in various organism and food, be mainly aquatic products.It plays an important role in allergic reaction, airway inflammation, nerve signal transmission and immunological regulation as a kind of important neurotransmitter.But when human body is taken the photograph too much, histamine poisoning (being also called mackerel poisoning) (Dickinson G.Scombroid fish poisoning syndrome [J] .Annals of EmergencyMedicine can be caused, 1982,11 (9): 487 – 489), cardinal symptom shows as uncomfortable in chest, headache, conjunctival congestion, vomiting and diarrhoea, nettle rash, hemorrhage of digestive tract, severe patient even threat to life (believes that system is gorgeous, Deng. edible mackerel causes histamine poisoning to investigate and analyse [J] together. Chinese Journal of Health Laboratory Technology, 2012,80 (7): 25-67.).At present, Conventional processing methods as heat treatment, microwave, ultrasonic etc. on histamine almost do not affect (Zhao Zhonghui, etc. different processing is on the research of biogenic amine impact. food industry science and technology, 2011,32 (7): 88-90).The method such as ice preservation, interpolation natural antiseptic agent is adopted to suppress to produce histamine microbial growth, histamine can be controlled and produce (Yang Jian, Deng. produce the progress [J] of histamine microorganism and control thereof in food. food industry science and technology, 2012,7 (3): 23-29).

Parvalbumin (parvalbumin) is the main allergen in fish muscle, and molecular weight is about 12.5kDa, is Ca ⁺in conjunction with water-solubility protein.People's Late Cambrian from cod (Gadus callarias) such as foreign scholar Elsayed in 1971, called after Gad c l (Ma Y, et al.Comparison of natural and recombinant forms of the majorfish allergen parvalbumin from cod and carp [J] .Mol Nutr Food Res, 2008,52 (1): 196 – 207).Follow-up domestic and international scientist conducts in-depth research other fish, and confirms that it is the main allergen in fish.Allergic symptom main manifestations is the symptoms such as allergic dermatitis, nettle rash, generalized patchy redness, asthma, oral cavity syndrome, enteropathy syndrome, and serious caused anaphylactic shock is threat to life even.There is certain difference in the medium and small albuminised content of fish different parts muscle, the medium and small albuminised content of the red meat comparatively low (Cai Qiufeng of white meat, Deng. the detection [J] of fish main allergen parvalbumin. China Immunology Journal, 2010,26 (8): 716-720.).In addition, the molecular weight that parvalbumin polymerization is formed is about the tetramer that the dimer of 24kDa or molecular weight be about 48kDa and also may reacts with IgG/IgE, has anaphylaxis.The hydrolysis, chemical modification etc. of parvalbumin to heat, protease have higher stability, therefore in process of manufacture, removing (Hamada Y is difficult to, et al.Expression and evaluation of IgE-binding capacity of recombinantPacific mackerel parvalbumin [J] .Allergol.Int, 2004,53 (3): 271 – 278).

China is global maximum fish production state.According to the statistics of China Fisheries yearbook, 2013, China's aquatic products total output reached 6,172 ten thousand tons, growth by 4.17% compared with 2012, and wherein marine product reaches 3138.86 ten thousand tons, accounts for 50.85% of total output.Containing enriching good protein and other nutrients in fish, deeply like by consumer.But can there is the problems such as the too high and parvalbumin of histamine content is irritated in consumer, thus limit consumer and obtain high-quality nutrition in fish, therefore, the processing method that research can reduce or remove these two kinds of materials has great importance when food fish.Blue circle Scad (Decapterus maruadsi) is one of China East Sea, South Sea Main Commercial Fishes, mainly be distributed in the Fujian coast in the Pacific Ocean and China Taiwan, the East Sea, its nutritive value is very high, the tender deliciousness of meat, protein content is up to 18.5%, fat content is only 3.4%, be rich in polyunsaturated fatty acid in fish body as several physiological active substances such as eicosapentaenoic acid (EPA), DHA (DHA), active peptides, its nutritive value can match in excellence or beauty with sardine.Blue circle Scad belongs to rascal red meat fish simultaneously, is rich in a large amount of histidine in body, easily decomposes under the effect of histamine bacterium and produces a large amount of histamine, causes flesh of fish histamine content to exceed standard.

Maillard reaction (MR) is the class complex chemical reaction that the material such as reduced sugar and amino acid, protein, amine occurs, and is also called carbonyl ammonia react.Maillard reaction is extensively present in food processing (as dried, frying, bake, boil, exploding), the local flavor of food and color and luster sense organ are had great importance (Li Yali, Deng. Maillard reaction progress [J]. food science and technology, 2012,37 (9): 82-87).Histamine is a kind of amine substance, can with glucose generation Maillard reaction, histamine content is reduced.Anaphylactogen is a kind of protein, can be changed or destroy the epitope of anaphylactogen, and then reduce its anaphylaxis by chemical modification.

Can processing have long shelf-life, simple to operate, easy to carry, the advantages such as the original color of food can be kept preferably, be the important trade aquatic product in the world, deeply like by consumer.

Summary of the invention

The object of this invention is to provide and can reduce blue circle Scad histamine content and parvalbumin anaphylaxis simultaneously, the processing method of technique advantages of simple, workable, easy large-scale production, a kind of indigo plant circle Scad can that product sensitization is low.

The present invention includes following steps:

1) after the reject indigo plant circle head of Scad, tail and internal organ, cleaning;

2) flesh of fish is flitched, cleaning, then impregnated in syrup;

3) flesh of fish flooded is put into the slack tank of having sterilized, add syrup, heat is steamed, and seals while hot, sterilization, cooling, vanning warehouse-in.

In step 1) in, indigo plant circle Scad can be carried out flowing water before the head of described reject indigo plant circle Scad, tail and internal organ and to thaw process, water temperature controls below 10 DEG C.

In step 2) in, the size of described fish block can be 1.5 ~ 2cm; Described cleaning can adopt clear water repeatedly to clean 3 ~ 5 times; Described syrup composition by mass percentage can be: reduced sugar 1% ~ 3%, salt 1% ~ 3%, and surplus is water; The time of described dipping can be 15 ~ 20min, and the temperature-controllable of dipping is built in less than 10 DEG C.

In step 3) in, described syrup composition by mass percentage can be: reduced sugar 1% ~ 3%, salt 1% ~ 3%, and surplus is water; The described syrup that adds can add to bottom clearance height 8 ~ 10mm; The time that described heat is steamed can be 15 ~ 25min; Described sterilization can adopt wet sterilization, and the temperature of sterilization can be 121 ~ 127 DEG C, the can standard that sterilizing time is issued with reference to national Ministry of Light Industry; Described cooling can adopt progressively cooling method to 35 ~ 40 DEG C.

The present invention adds syrup in indigo plant circle Scad can processing process, utilizes reduced sugar in the processes such as heat steaming, Maillard reaction to occur, simultaneously in conjunction with the destruction etc. of HTHP to parvalbumin space structure with histamine in the flesh of fish and parvalbumin.

Advantage of the present invention is: add syrup in blue circle Scad can process of manufacture; reduced sugar is utilized in the processes such as heat steaming, Maillard reaction to occur with the histamine in the flesh of fish and parvalbumin; utilize high temperature high pressure process to methods such as the destructions of the medium and small albumin structure of the flesh of fish simultaneously; while keeping the blue circle peculiar quality of Scad and nutrition to greatest extent; cut down anaphylaxis material histamine and parvalbumin, there is the features such as easy and simple to handle, cost is low, easily accomplish scale production.

Accompanying drawing explanation

Fig. 1 be in Maillard reaction different glucose on the impact of histamine content.

Fig. 2 be in Maillard reaction different glucose on the impact of parvalbumin.In fig. 2, PV, parvalbumin; G, glucose; Con, control group is oppressed.

Fig. 3 is the impact of high temperature high pressure process different time on parvalbumin IgE binding activities.

Fig. 4 is the indigo plant circle Scad can that embodiment 4 processes.In the diagram, the indigo plant circle Scad flesh of fish of A. sugaring (1% glucose, 2% salt); B. the indigo plant circle Scad flesh of fish after the steaming of syrup heat is added; C. the indigo plant circle Scad can after high temperature high pressure process.

Fig. 5 is the impact of the different processing mode of embodiment 5 on histamine content in the indigo plant circle Scad flesh of fish.In Figure 5,1, contrast; 2, sugaring; 3, heat is steamed; 4, high temperature high pressure sterilizing process; 5, Maillard reaction is in conjunction with can processing.

Fig. 6 is that the different processing mode of embodiment 6 oppresses the impact of medium and small albuminised IgE binding activities to indigo plant circle Scad.In figure 6,1, contrast; 2, sugaring; 3, heat is steamed; 4, high temperature high pressure sterilizing process; 5, Maillard reaction is in conjunction with can processing.

Fig. 7 is that before and after embodiment 7 is processed, in the blue circle Scad flesh of fish, parvalbumin activates rat basophilic granulocyte (RBL-2H ₃) release β-hexosaminidase impact.In the figure 7,1, PBS; 2, contrast; 3, sugaring; 4, heat is steamed; 5, HTHP; 6, Maillard reaction is in conjunction with can processing.

Detailed description of the invention

Below in conjunction with the drawings and specific embodiments, the present invention is further illustrated.

Embodiment 1: in Maillard reaction, the glucose of variable concentrations is on the impact of histamine content

(1) preparation of glucose solution and histamine standard liquid: configuration 1%, 2%, 3%, 4% glucose solution is the histamine standard liquid of 200 μ g/mL with histamine dihydrochloric acid standard items compound concentration simultaneously.

(2) set up Maillard reaction system: by glucose solution and histamine standard liquid according to 1: 1 volume ratio mix, make histamine standard liquid final concentration be that to be adjusted to pH be 9.5 for 100 μ g/mL, 0.1M NaOH.

(3) Maillard reaction of glucose and histamine standard items: ready sample is positioned in 80 DEG C of water-baths and reacts 30min, with the histamine standard liquid of 100 μ g/mL in contrast, glucose variable concentrations is detected in Maillard reaction on the impact of histamine standard items content.

Embodiment 2: in Maillard reaction, the glucose of variable concentrations is to the IgE binding activities of parvalbumin

(1) Maillard reaction system is set up: the solution (3: 1,2: 1,1: 1,1: 3,1: 5,1: 7,1: 9) parvalbumin and glucose being mixed with different quality ratio, make parvalbumin final concentration be 0.5 μ g/mL, being adjusted to pH with 0.1M NaOH is 9.5.

(2) Maillard reaction of glucose and parvalbumin: sample is positioned in 80 DEG C of water-baths and reacts 30min.

(3) Dot-blotting analyzes: with reference to Towbin method (Towbin H, et al.Electrophoretic transferof proteins from polyacrylamide gels to nitrocellulose sheets:Procedure and someapplications.Proceedings of the National Academy of Sciences of the United Statesof America, 1979,76 (9): 435-4354) the IgE binding activities that Dot-blotting analyzes parvalbumin after Maillard reaction is carried out.The direct point sample of protein sample is on nitrocellulose filter (NC) film, with 1% defatted milk dilution fish allergic patients sera (1: 4) be primary antibodie, the sheep anti human IgE that horseradish peroxidase (HRP) marks be two resist, using diaminobenzidine as nitrite ion, develop the color and take pictures.

Embodiment 3: high temperature high pressure process different time is analyzed the IgE binding activities of parvalbumin

(1) high temperature high pressure process: be 0.5 μ g/mL by parvalbumin Concentration Modulation, autoclave sterilization pot temperature is set to 121 DEG C, and pressure is 0.12MPa, sterilizing time is respectively 5,10,15,20,25min.

(2) Dot-blotting analyzes in reference example 2 (3).

Embodiment 4: reduce histamine content and the hypersensitive processing method of parvalbumin in blue circle Scad can, comprise the following steps:

(1) pretreatment of raw material: indigo plant circle Scad is carried out flowing water and to thaw process (water temperature controls below 10 DEG C), the head of reject fish, tail and internal organ, with foreign material such as clear water flushing clean Intraabdominal blood stains, black films;

(2) sugaring: the fish block flesh of fish being cut into the even size of 1.5cm, clear water cleans 3 times repeatedly; The fish block cut impregnated in 20min (soaking temperature controls below 10 DEG C) in syrup (1% reduced sugar and 2% salt);

(3) heat is steamed and exhaust: the soaked flesh of fish is put into the slack tank of having sterilized, add syrup (1% reduced sugar and 12% salt) to bottom clearance height 10mm, and heat steams 20min, seals while hot;

(4) high temperature high pressure sterilizing: adopt high temperature and high pressure steam sterilization, sterilization temperature is 121 DEG C, sterilizing time 20min;

(5) progressively cool, warehouse-in of casing: adopt stepwise process be cooled to about 40 DEG C, dry tank table moisture, vanning warehouse-in.

Embodiment 5: histamine in the pre column Derivatization indigo plant circle Scad flesh of fish

(1) derivatization of histamine standard liquid: with histamine dihydrochloric acid standard items respectively compound concentration be the standard liquid of 1,5,10,20,50,100,200,500 μ g/mL.Accurate absorption 300 μ L sample solution is in 2mL centrifuge tube, and NaOH is adjusted to alkalescence.In solution, add 600 μ L/mL Dns-Cl solution, after 40 DEG C of water-bath 20min terminate, add 100 μ g/mL ammoniacal liquor mixings, after lucifuge leaves standstill 30min, be settled to 1.5mL with acetonitrile and cross 0.22 μm of organic film, upper machine is to be measured.

(2) extraction of sample: accurately take 5.0g sample in 50mL centrifuge tube, adds 15mL 5% trichloroacetic acid and smashs homogenate to pieces, 8000r/min, 4 DEG C, centrifugal 15min, and supernatant liquor moves in the brown volumetric flask of 25mL.Sediment adds 10mL 5% trichloroacetic acid again and repeats extraction 1 time according to aforesaid operations, and supernatant is incorporated in volumetric flask, is settled to 25mL scale with 5% trichloroacetic acid.

(3) derivatization of sample: add 40 μ L 2mol/L sodium hydroxide solutions in 300 μ L sample extracting solutions, adjusts about pH to 9.5, then adds 600 μ L 10mg/mL dansyl chloride solution, after mixing, in 40 DEG C of thermostat water baths, react 20min.After completion of the reaction, add 100 μ L ammoniacal liquor, mixing, leave standstill 30min.Be settled to 1.5mL with acetonitrile, after vibration mixing, 10000r/min, 4 DEG C, centrifugal 5min, supernatant is to be measured after crossing the organic phase pin type filter of 0.22 μm.

(4) column front derivation high-efficient liquid phase chromatogram condition chromatographic column: Agilent ZORBAX Extend-C18 reverse-phase chromatographic column (4.6mm I.D. × 150mm, 5 μm); Mobile phase: acetonitrile: water=70:30 (V/V); Column temperature: 35 DEG C; UV detect wavelength: 254nm; Flow velocity: 1mL/min; Sample size: 20 μ L.

Embodiment 6: before and after processing, blue circle Scad oppresses the mensuration of medium and small albuminised IgE binding activities

(1) slightly carrying of parvalbumin is oppressed: will oppress and the mixing of soup juice, and smash to pieces (low-temperature operation) with tissue mashing machine, 12000g, 4 DEG C, centrifugal 20min, gets supernatant four layers of silk and filters, except the impurity such as degrease, insoluble protein, supernatant is for subsequent use.

(2) mensuration of medium and small albuminised IgE binding activities is oppressed: enzyme linked immunosorbent assay (ELISA) (ELISA): adopt the IgE binding activities of inhibition ELISA to sample to analyze.Diluted by parvalbumin crude extract coating buffer, wrap by 96 hole ELISA Plates, every hole bag is by 200ng, and 100 μ L/ holes, place 16h for 4 DEG C.Liquid in hole inclines by next day, pats dry, and cleans 5 times with TBST.Then 5% defatted milk is added in 37 DEG C of closed 2h.Allergen protein elaboration products (the sugaring of 30 μ L fish allergic patients sera (1: 4 with 1% defatted milk dilution) and equivalent is got respectively in this external centrifuge tube, heat is steamed, high temperature high pressure sterilizing process, can processing) and undressed allergen protein (control group) mixing hatch, allergen protein elaboration products, as inhibitor, hatch 1h at 37 DEG C.End to be closed, after TBST cleaning of enzyme target, joins in hole by completely reacted mixed liquor with 50 μ L/ holes, 37 DEG C, continues to hatch 1h.With TBST washing, add the sheep anti human IgE two anti-(1: 2000) that 100 μ L HRP mark, 37 DEG C, hatch 2h.Tetramethyl benzidine develops the color, and color display is blue.Add 2mol/L sulfuric acid solution cessation reaction, color display is yellow.Light absorption value under ELIASA detection 450nm.The calculating of inhibiting rate: inhibiting rate (%)=(X-Y) ÷ (X-Z) × 100, wherein X is the light absorption value that allergic patients sera does not have inhibiting, Y and Z is that the light absorption value after hatching competed by positive serum and negative serum and inhibitor respectively.

Embodiment 7: before and after processing, in the flesh of fish, parvalbumin activates the impact of rat basophilic granulocyte release β-hexosaminidase

(1) basophilic granulocyte is cultivated: will be stored in the rat basophilic granulocyte (RBL-2H in liquid nitrogen container ₃) carry out cell recovery cultivation, add containing 10% hyclone and 1% dual anti-MEM/EBSS culture medium 2mL in each culture dish, cultivate attached cell for 37 DEG C.Every 2d changes a not good liquor, observes counting at any time, and final adjustment cell density is 1 × 10 ⁶/ mL, collecting cell.

(2) oppress elaboration products and activate basophilic granulocyte situation: be added to after flesh of fish crude extract protein concentration is diluted to 1 μ g/mL in 6 porocyte culture plates, 20 μ L mouse specific serums are added in every hole, after sensitized overnight, 4 DEG C, 1200g, 10min are centrifugal, wash 1 time with PBS.Add Tyrode ' s buffer (200 μ L/well), 37 DEG C, hatch 6h.Get supernatant respectively with precipitation, after adding 200 μ L 0.1%triton X100 in precipitation, above-mentioned cell (supernatant is with the cell after lysis) is added M2133 retting conditions reagent, 37 DEG C, reaction 30min.Get 200 μ L and add 2mL0.1M sodium acetate cessation reaction, ELIASA 405nm place surveys light absorption value.Calculate β-hexosaminidase release rate as follows:

β-hexosaminidase release rate=cell conditioned medium β-hexosaminidase content/(cell conditioned medium β-hexosaminidase content+cell pyrolysis liquid β-hexosaminidase content) × 100%.

Fish block be impregnated in syrup, alleviate the peculiar smell of fish body on the one hand, suppress growth of microorganism; Make syrup immerse in structure of fish muscle on the other hand, be convenient to, in subsequent process, Maillard reaction occurs.

Add syrup in can, the flesh of fish environment heat that is placed in reduced sugar is steamed, impel fish block musculature closely, while texture characteristic improves, be convenient to the flesh of fish and in the processes such as heat steaming, Maillard reaction occur with reduced sugar solution.

The flesh of fish belongs to low acid can, and HTHP is convenient to kill the bacteriums such as clostridium botulinum, simultaneously the space structure of broken ring parvalbumin, realizes the object of abatement flesh of fish anaphylaxis material.

Adopt progressively cooling method, prevent Cans to be easy to when temperature sharply changes burst.

Claims (10)

1. a processing method for blue circle Scad can, is characterized in that comprising the following steps:

1) after the reject indigo plant circle head of Scad, tail and internal organ, cleaning;

2) flesh of fish is flitched, cleaning, then impregnated in syrup;

3) flesh of fish flooded is put into the slack tank of having sterilized, add syrup, heat is steamed, and seals while hot, sterilization, cooling, vanning warehouse-in.

2. the processing method of a kind of indigo plant circle Scad can as claimed in claim 1, is characterized in that in step 1) in, before the head of described reject indigo plant circle Scad, tail and internal organ, indigo plant circle Scad is carried out flowing water and to thaw process, water temperature controls below 10 DEG C.

3. a kind of indigo plant justifies the processing method of Scad can as claimed in claim 1, it is characterized in that in step 2) in, the size of described fish block is 1.5 ~ 2cm.

4. a kind of indigo plant justifies the processing method of Scad can as claimed in claim 1, it is characterized in that in step 2) in, described cleaning adopts clear water repeatedly to clean 3 ~ 5 times.

5. the processing method of a kind of indigo plant circle Scad can as claimed in claim 1, is characterized in that in step 2) and 3) in, described syrup consisting of by mass percentage: reduced sugar 1% ~ 3%, salt 1% ~ 3%, surplus is water.

6. a kind of indigo plant justifies the processing method of Scad can as claimed in claim 1, it is characterized in that in step 2) in, the time of described dipping is 15 ~ 20min, and the temperature of dipping controls below 10 DEG C.

7. the processing method of a kind of indigo plant circle Scad can as claimed in claim 1, is characterized in that in step 3) in, described in add syrup be add to bottom clearance height 8 ~ 10mm.

8. a kind of indigo plant justifies the processing method of Scad can as claimed in claim 1, it is characterized in that in step 3) in, the time that described heat is steamed is 15 ~ 25min.

9. a kind of indigo plant justifies the processing method of Scad can as claimed in claim 1, it is characterized in that in step 3) in, described sterilization adopts wet sterilization, and the temperature of sterilization is 121 ~ 127 DEG C, the can standard that sterilizing time is issued with reference to national Ministry of Light Industry.

10. a kind of indigo plant justifies the processing method of Scad can as claimed in claim 1, it is characterized in that in step 3) in, described cooling adopts progressively cooling method to 35 ~ 40 DEG C.