CN103053666B - Method for determining best ice temperature cold induction mode of fish flesh - Google Patents
Method for determining best ice temperature cold induction mode of fish flesh Download PDFInfo
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Abstract
The invention provides a method for maintaining the fish flesh freshness maintaining and the fish flesh taste increasing through ice temperature cold induction. The freshness maintaining and taste increasing method comprises the following steps: 1, assaying the ice point of fish flesh; 2, determining the lethal temperature of fish; 3, treatment before live fish cold induction: temporarily rearing live fish in water having a temperature 3-5DEG C higher than the lethal temperature for above 1h, killing the fish in an environment having a temperature 3-5DEG C higher than the lethal temperature, removing internal organs, and allowing the fish to be complete, or segmenting or slicing; and 4, cold induction: cooling the fish flesh to an ice temperature range, and storing at a temperature in the ice temperature range for a period of time. The invention also provides a method for determining the best ice temperature cold induction mode of the fish flesh. The best cold induction mode is determined through assaying the freshness index K value and the taste component based on the freshness maintaining and taste increasing method, and evaluating the freshness and the taste of the fish flesh undergoing different cold induction modes. The methods allow fresh and delicious fish flesh to be obtained, provide references for the fish flesh storing and processing mode selection, and are of great guiding significances to the improvement of the fish selling quality, especially the maintenance of the freshness of processed aquatic products.
Description
Technical field
The present invention relates to food preservative technology field, specifically, is the method for the cold induction mode of the best ice temperature of a kind of definite flesh of fish.
Background technology
Tilapia mossambica (Tilapias) claim again African crucian, belong to Perciformes (Perciformes) Li Diao section (Cichlidae) Tilapia mossambica and belong to (Tilapia), because its food polygamy, strong adaptability, breeding potential are high, the main cultured fishes that become international by force of fast growth, disease resistance, also become the important freshwater aquiculture object of China.Because Tilapia mossambica is higher to water temperature requirement, existence temperature range is 15 ℃ ~ 35 ℃, larger in the higher area cultivation of China's Guangdong equitemperature.Tilapia mossambica is the cultured freshwater fish that the emphasis scientific research of World Water industry is cultivated, and is described as one of main source of following animal protein.
The fish such as Tilapia mossambica through catch lethal after, in its body, still carrying out the variation of various complexity, by just freshness is after death good in putrid and deteriorated, to pass through substantially stiff, self-dissolving and putrid and deteriorated three phases.Conventionally the seafood degree before stiff or in stiff is good, and the autolysis stage freshness quality of fish starts to decline, and when protein is decomposed by enzyme, the material of amino acid one class increases, for the breeding of bacterium provides condition.
Utilize adenosine triphosphate in fish muscle (ATP) after death the initial stage decomposes, through adenosine diphosphate (ADP), adenylate (AMP), inosinicacid (IMP), inosine (HxR), hypoxanthine (Hx) etc., the wherein ratio of (HxR+Hx) and ATP and catabolite summation thereof, is index of fish freshness K value:
General K value≤20% that adopts is as good index of fish freshness (Japan is for the quality standard of fresh fish), and K value≤60% is as the freshness standard processing raw material.
Generally believe that IMP is the main component of delicate flavour, the generation of IMP can improve the delicate flavour of fish, and in the time that IMP concentration reduces, the local flavor of fish just becomes unacceptable gradually.
Free amino acid is the main component of non-volatile nitrogenous thing in taste substance, in the time that some free amino acid is present in the muscle of fish with sufficiently high concentration, can be independent of other compositions the local flavor of fish is worked.As glycine, alanine produce sweet taste, lysine produces sweetness and bitterness taste, and histidine forms " the meat perfume (or spice) " of some marine product, and glutamic acid, aspartic acid and alanine etc. have larger contribution to delicate flavour.
Extend preservation time, the maintenance fish freshness of the aquatic products such as Tilapia mossambica and increase flesh of fish flavour for improving fish market quality, especially keeping process water product freshness significant.
At preservation of fishery technical elements, domestic and international existing document is mainly to concentrate on iced storage and cold storing and fresh-keeping, and that the at present research of ice-temperature technique is reported is at home also more and more.Ice temperature refers to that 0 ℃ starts to the temperature province of organism chill point, and Icetemperature Storage refers to the storage in this temperature band.The outstanding advantages of Icetemperature Storage compared with cold storage is not destroy cell; The outstanding advantages of ice temperature compared with refrigeration is that not only harmful microbe activity movable and various enzymes is suppressed, and extends storage period, and can improve the quality of food.To black carp (Liang Qiong, Wan Jinqing, Wang Guoqiang. black carp sheet Icetemperature Storage research [J]. Food Science, 2010 (06): 270-273.), lefteye flounder (Li Hui, Liu Lianfeng, Yang Bofeng etc. the freshness of lefteye flounder and texture change [J] under ice temperature fresh-keeping condition .) etc. research show, Icetemperature Storage condition can more effectively suppress the breeding of aquatic products internal microorganism, extends storage period of food.
In recent years, Japanese scholars in ice-temperature technique aspect two of lay special stress ons (take the Country man of virtue and ability of portion. improve the ice-temperature technique [C] of food freshness and taste .), the one, the importance of " organism defense reaction ", the 2nd, prevent temperature change technology, i.e. constant temperature technology.The former says, is placed on the fresh food on dining table, from cell angle, can think that they are lived.The dead fish of processing take the internal organ of eating at home is example, the temperature of fish is slowly reduced to ice Wen Wendu band, gradually the sensation artificially of closing on winter is passed to it, along with approaching solidification point, as fish self precognition will be freezed to death, the concentration of non freezing solution in cell (as amino acid etc.) increases, and through the regular hour, taste can become deliciousness.In addition, because Icetemperature Storage temperature field is in low-temperature space, harmful microorganism and pathogenic bacteria significantly reduce, and this temperature field is also the very active field of enzyme, yeast and lactic acid bacteria of improving mouthfeel and local flavor, therefore, need not add anticorrisive agent.
But, neither say that any fresh food or food material can increase due to the effect of organism defense reaction and enzyme the delicious food of food.By promoting the defense reaction of organism, improving its effect is one of feature of ice-temperature technique.For different food materials, in order organism defense reaction to be amino acid whose conversion maximization, the guiding of its chilling temperature is different, this is that ice temperature effect is not easy the maximum reason realizing: conventionally, front to higher temperature (10 ~ 5 ℃ of left and right) for guiding temperature, i.e. use is cooling fast, as also not giving the impact large with ice temperature effect.But, the speed declining from later temperature, will be very slow, can not be irritable, carry out so-called domestication by low temperature operation, gradually the sensation artificially of closing on winter is passed to natural biology, allow them start the preparation that arrive winter, cold resistance that its result has made bio-augmentation from physiology.But, for different foodstuffs, not only to lower the temperature in certain proportion, during the course, the hopping time that a gap also will be set seems effective means.In addition, as some citrus, entering before low temperature field, the temperature band below+5 ℃, the fruit and vegetable based food that has some may cause low temperature infringement, this situation will be found the countermeasure of eliminating low temperature infringement simultaneously.
Chinese periodical " Food Science ", 06 phase in 2010, the paper " research of black carp sheet Icetemperature Storage " of publishing, is placed on black carp sheet respectively under the environment of (0.8 ± 0.2), (2.0 ± 1.0) ℃ and (4.0 ± 1.5) ℃ and preserves, and regularly takes out and measures its sense organ, microorganism and physical and chemical index.In the storage end of term, the sample total plate count under 3 kinds of holding conditions is respectively 1.17 × 10
6(the 11st day), 1.12 × 10
6cFU/g(the 8th day) and 1.2 × 10
6cFU/g(the 5th day); TVB-N value is respectively 24.95(the 20th day), 20.03mg/100g(the 15th day) and 20.39mg/100g(the 12nd day); PH value is respectively 6.96(the 12nd day), 6.59(the 7th day) and 6.73(the 8th day).Result shows, black carp sheet refrigeration and micro-approached while freezing under condition respectively the 5th day of experiment and the 8th day corrupt, and in the black carp sheet of Icetemperature Storage approaching corruption just during at the 11st day.
Chinese periodical " fishery science progress ", 03 phase in 2011, the paper " freshness of lefteye flounder and texture change under ice temperature fresh-keeping condition " of publishing, the index of fish freshness (pH value, K value, TVB-N, total number of bacteria) that research lefteye flounder is oppressed under Icetemperature Storage condition and the variation of matter structure characteristic parameter (subjective appreciation, histological structure, rupture strength etc.), and compare analysis with the respective change of refrigeration sample.Result shows, along with the prolongation of storage time, total plate count, K value and TVB-N rise, and pH value first declines and rises afterwards, refrigerates later stage sample pH value value rising very fast and very high; During to shelf life terminal, rupture strength is and reduces trend, and institutional framework is deteriorated gradually.Compared with refrigeration sample, Icetemperature Storage condition can more effectively suppress the effect of lefteye flounder fish internal microorganism, extends the storage period of lefteye flounder.
The ice temperature method of above-mentioned aquatic products is all directly fresh sample to be put in to storage in ice warm area, and have not been reported with the method that keeps fish freshness and increase flesh of fish flavour about ice temperature after cold induction at present, and effectively determine that the method for the cold induction mode of the best ice temperature of the flesh of fish also there is not yet report.
Summary of the invention
The object of the invention is for deficiency of the prior art, provide the cold induction of a kind of ice temperature to keep fish freshness and increase the method for oppressing flavour.
Second object of the present invention is that the method for the cold induction mode of the best ice temperature of a kind of definite flesh of fish is provided.
For achieving the above object, the technical scheme that the present invention takes is:
The cold induction of ice temperature keeps fish freshness and increases a method for flesh of fish flavour, and it comprises the following steps:
A) flesh of fish freezing test;
B) determine the lethal temperature of fish;
C) processing before the cold induction of live fish: live fish more than temporary foster 1h, is slaughtered fish in the water higher than 3～5 ℃ of lethal temperatures in the environment higher than 3～5 ℃ of lethal temperatures, gilled, whole piece, segmentation or section;
D) cold induction: flesh of fish temperature is down to behind its ice temperate zone, in ice temperate zone storage a period of time.
Its concrete way of flesh of fish freezing test described in step a) is: fish is slaughtered, gill, clean up, by thermoelectricity occasionally thermal resistance equitemperature sensor insert under fish body surface about 0.5cm place and fix, put into the cold storage chamber of-20 ℃, temperature acquisition is spaced apart 10s and gathers a secondary data, and experiment finishes rear drafting freezing curve and draws the freezing point of the flesh of fish.
Its concrete way of the lethal temperature of definite fish described in step b) is: live fish is placed in ecological tank and is supported temporarily, with the speed of 1 ℃/24h to water for cooling, death time and corresponding water temperature situation are once recorded in every 2h manual observation, the death standard of check experiment fish is, the gill cover stop agitating and acupuncture reactionless, after experiment fish is placed in to natural temperature, in 5 minutes, do not reactivate.3 repeated experiments, chooses the maximum temperature while there is dead fish, as the lethal temperature of fish.
For realizing above-mentioned second object, the technical scheme that the present invention takes is:
A method for the cold induction mode of the best ice temperature of definite flesh of fish, it comprises the following steps:
A) flesh of fish freezing test;
B) determine the lethal temperature of fish;
C) processing before the cold induction of live fish: live fish more than temporary foster 1h, is slaughtered fish in the water higher than 3～5 ℃ of lethal temperatures in the environment higher than 3～5 ℃ of lethal temperatures, gilled, whole piece, segmentation or section;
D) cold induction: the flesh of fish is put into the cold environment induction of lowering the temperature, and the finishing temperature of oppressing cold induction is its ice temperate zone, and the flesh of fish is preserved to a period of time in ice temperate zone;
E) measure index of fish freshness K value and flavour composition, the flesh of fish of the cold induction mode of difference is carried out to the evaluation of freshness and flavour, determine best cold induction mode.
Its concrete way of flesh of fish freezing test described in step a) is: fish is slaughtered, gill, clean up, by occasionally thermal resistance equitemperature sensor of thermoelectricity, insert about 0.5cm place fixing under fish body surface, put into the cold storage chamber of-20 ℃, temperature acquisition is spaced apart 10s and gathers a secondary data, and experiment finishes rear drafting freezing curve and draws the freezing point of the flesh of fish.
Its concrete way of the lethal temperature of definite fish described in step b) is: live fish is placed in ecological tank and is supported temporarily, with the speed of 1 ℃/24h to water for cooling, death time and corresponding water temperature situation are once recorded in every 2h manual observation, the death standard of check experiment fish is, the gill cover stop agitating and acupuncture reactionless, after experiment fish is placed in to natural temperature, in 5 minutes, do not reactivate, 3 repeated experiments, chooses the maximum temperature while there is dead fish, as the lethal temperature of fish.
The invention has the advantages that:
1, the invention provides the method that the cold induction of a kind of ice temperature keeps fish freshness and increases flesh of fish flavour, can obtain the flesh of fish of flavour deliciousness;
2, the cold induction of ice temperature of the present invention keeps fish freshness and increases the method for oppressing flavour, by live fish in the water higher than 3～5 ℃ of lethal temperatures more than temporary foster 1h, in the environment higher than 3～5 ℃ of lethal temperatures, fish is slaughtered, to maintaining the flavour composition of the flesh of fish, guarantee that the conversion of flavour composition in the cold Induction Process of ice temperature has important function;
3, known to different food materials, in order organism defense reaction to be amino acid whose conversion maximization, the guiding of its chilling temperature is different, this is that ice temperature effect is not easy the maximum reason realizing, the initial cold inducing temperature that the present invention carries out cold induction to Tilapia mossambica adopts 15 ℃, can obtain good cold induction effect;
4, the method for the cold induction best mode of definite flesh of fish ice temperature of the present invention is preservation time of extending the aquatic products such as Tilapia mossambica, keeps its freshness and increase its flavour, for improving fish market quality, especially keep process water product freshness to there is important directive significance.
Accompanying drawing explanation
Accompanying drawing 1 is Tilapia mossambica freezing curve.
Accompanying drawing 3 is standard items HPLC collection of illustrative plates of ATP and catabolite thereof.
Accompanying drawing 4 is that under different cold inductive condition, Tilapia Fillet K value changes.
Accompanying drawing 5 is the variations of Tilapia mossambica IMP and ATP under different cold inductive conditions.
Accompanying drawing 6 is 35h temperature lowering curves before Tilapia mossambica fillet.
Accompanying drawing 7 is that under 5 kinds of cold inductive conditions of different initial temperatures, Tilapia Fillet K value changes.
Accompanying drawing 8 is the variations of Tilapia mossambica IMP and ATP under 5 kinds of cold inductive conditions of different initial temperatures.
The specific embodiment
Below in conjunction with accompanying drawing, the specific embodiment provided by the invention is elaborated.
1 materials and methods
1.1 material
Tilapia mossambica: purchase in food market, Lingang New City Gu Zong road, Shanghai City, body weight 500 ~ 600g/ tail, after buying, laboratory is transported rapidly in oxygenation back.
1.2 reagent and equipment
Reagent: standard items comprise atriphos (Adenosine Triphosphate, and compound adenosine diphosphate (ADP) (Adenosine Diphosphate ATP), ADP), adenosine monophosphate (Adenosine Monophosphate, AMP), inosinicacid (Inosine Monophosphate, IMP), inosine (Inosine, HxR), hypoxanthine (Hypoxanthine, Hx), wherein ATP, ADP, IMP, Hx standard items are produced by Sigma company, AMP standard items are produced by Japanese TCI company, HxR standard items are produced by German Dr. Ehrenstorfer company.Methyl alcohol is HPLC level (Chemical Reagent Co., Ltd., Sinopharm Group); Phosphate HPLC level (Town in Shanghai spectrum scientific instrument company); Experimental water all adopts ultra-pure water; It is pure that perchloric acid (PCA), potassium hydroxide, NaOH, phosphoric acid, sulfosalicylic acid are analysis.
Equipment: Japanese Shimadzu LC-2010CHT high performance liquid chromatograph; HITACHI L-8800 amino acid fully-automatic analyzer; Agilent 34972A Agilent temperature sensor; Shanghai Yiheng Scientific Instruments Co., Ltd's climatic chamber BPS-100CL(-10 ~ 100 ℃), BPS-250CB (40 ~ 100 ℃), LHS-100CA (20 ~ 35 ℃); The full-automatic snowflake ice machine of Changshu-Xue Ke IMS-50; Ningbo new sesame biotechnology SB25-12DT ultrasonic machine; Fluko FA25 laboratory high-shear homogenizing machine; Japan Shimadzu AUW320 electronic balance; Thunder magnetic PHS-3C type pH meter; Shanghai one permanent scientific instrument DHG-9053A electric drying oven with forced convection; Hunan, Changsha instrument H2050R refrigerated centrifuge; GM-0.33A vacuum diaphragm pump and solvent filter are risen in Tianjin; Qingdao Haier BCD-216SCM refrigerator.
The equipment of experiment use all adopts ultrasonic machine to clean 20min, repeatedly rinses with distilled water, then uses ultra-pure water rinse 2 ~ 3 times, and drying machine drying is for subsequent use.
1.3 test method
1.3.1 oppress freezing test
Along vertebra, fish being cutd open is two, and thermocouple is inserted to about 0.5cm place fixing under fish body surface, puts into the cold storage chamber of-20 ℃, and temperature acquisition is spaced apart 10s and gathers a secondary data, and experiment finishes rear drafting freezing curve and draws Tilapia mossambica freezing point.
1.3.2 determine the lethal temperature of fish
10 tail live fishes are placed in ecological tank and are supported temporarily, with the speed of 1 ℃/24h to water for cooling, death time and corresponding water temperature situation are once recorded in every 2h manual observation, the death standard of check experiment fish is, the gill cover stop agitating and acupuncture reactionless, after experiment fish is placed in to natural temperature, in 5 minutes, do not reactivate.3 repeated experiments, chooses the maximum temperature while there is dead fish, as the lethal temperature of fish.
1.3.3 the processing before the cold induction of live fish
Live fish is supported temporarily in the water higher than 3～5 ℃ of lethal temperatures to 1 ~ 2h, in the environment higher than 3～5 ℃ of lethal temperatures, fish is slaughtered, gill, sheet becomes fillet, is distributed into freshness protection package and is placed in cold environment.
1.3.4 cold induction---the setting of different cold induction mode cooling process
3 kinds of different cold induction modes, temperature control continuous cooling (mode I), the cooling of temperature control ladder (mode II), direct ice temperature drop temperature (mode III), used respectively 3 climatic chambers.Initial cooling temperature has all adopted 15 ℃, to guarantee that Tilapia mossambica keeps fresh and alive at this temperature.
Mode I, is first stabilized in climatic chamber 15 ℃ of lasting 2h, and fillet are placed in to steady-state zone, casing central authorities temperature field, and cooling process is, per hourly between 15 ℃ → 1 ℃ falls 1 ℃, per hourly between 1 ℃ →-0.5 ℃ falls 0.5 ℃; Allow the flesh of fish adapt to slowly cold environment, until be down to ice temperate zone, and maintain ice temperature condition a period of time.
Mode II, first climatic chamber is stabilized in to 15 ℃ of lasting 2h, fillet are placed in to steady-state zone, casing central authorities temperature field, cooling process is, directly be down to 10 ℃ and holding temperature from 15 ℃, after 5h, (since 15 ℃ of frigorimeters time) is directly down to 4 ℃ and keep temperature-resistant again, and after 10h, (since 10 ℃ of frigorimeters time) is directly down to-0.5 ℃ and maintain ice temperature condition a period of time again.
Mode III, is first stabilized in climatic chamber-0.5 ℃ of lasting 2h, fillet is put into, and maintained ice temperature condition a period of time.
1.3.5 the detection of ATP and relationship compound thereof
With reference to Yokoyama(Yokoyama Y, Sakaguchi M, et al. Change in concentration of ATP-related compounds in various tissues of oyster during ice storage[J]. Nippon Suisan Gakkaishi, 1992,58(11): 2125-2136.) and (QIU WEIQIANG such as QIU WEIQIANG, Chen Gang, Chen Shunsheng etc. ion pair reversed-phased high performace liquid chromatographic detects 6 kinds of ATP relationship compounds [J] in aquatic products simultaneously. aquatic product journal, 2011, (11) method: 1745-1752.), slightly changes.
(1) extraction of fillet ATP and relationship compound (ADP, AMP, IMP, HxR, Hx)
Get fresh fillet and the sample after cold treatment, after chopping, get 5g fast and put into centrifuge tube, add perchloric acid (PCA) the solution making beating 2min of the 10ml 10% of precooling.After homogenate, with 10000r/min refrigerated centrifuge 15min, get supernatant.5% PCA washing of precooling for precipitation, centrifuging and taking supernatant, repeats once.Merge supernatant, first with after 10mol/L KOH solution with 1mol/L KOH solution adjust pH to 6.5, supernatant is transferred in 50mL volumetric flask after leaving standstill 30min, and uses ultra-pure water constant volume, shake up, with to be determined after 0.45 μ m micro porous filtration membrane filtration etc.Whole process all operates under 0 ~ 4 ℃ of condition.
(2) high performance liquid chromatography (HPLC) detects chromatographic condition
Chromatogram (HPLC) condition: the Inertsil ODS-SP C18(4.6 × 250mm of GL Sciences company, m) liquid-phase chromatographic column of 5 μ; Guard column post core Inertsil ODS-SP(4 × 10mm, 5 μ are m); Mobile phase: A is 0.05mol/L potassium dihydrogen phosphate and dipotassium hydrogen phosphate (1:1) solution, and being adjusted to pH with phosphoric acid is that 6.5, B is methanol solution; Isocratic elution; Flow velocity: 1mL/min; Column temperature: 28 ℃; Sample size: 10 μ L; Check wavelength: 254nm.
1.3.6 the detection of free amino acid
With reference to Deng Jiechun (Deng Jiechun, Wang Xichang, Liu Yuan. fugu obscurus and Fugu rubripes flavour component difference research [J]. food industry science and technology, 2010,: 106-108.) and Xue Song (Xue Song (3), Wan Jinqing. the impact [J] of Icetemperature Storage on chicken freshness and free amino acid variation. Jiangsu agricultural sciences, 2010 (6): 411-413.) free amino acid assay method, slightly change.
(1) extraction of free amino acid
Take respectively flesh of fish 2g, add 10mL 5%(w/v) sulfosalicylic acid, precipitation 2h gets off protein precipitation, draw the centrifugal 15min of 6mL supernatant 10000r/min refrigerated centrifuge, get 3mL supernatant, with a certain amount of NaOH solution modulation pH to 2.0 left and right, be settled to 12mL, measure with being filled to upper machine in sample disc after 0.45 μ m micro porous filtration.
(2) analysis condition
L-8800 type automatic amino acid analyzer, sample analysis cycle 53min.
Chromatographic column: 4.6 × 150mm, 7 μ m;
Column temperature: 50 ℃;
Flow velocity 1:0.4ml/min, flow velocity 2:0.35ml/min;
Mobile phase: the buffer solution of citric acid, natrium citricum and ninhydrin.
2 interpretations
2.1 freezing test
The freezing curve of Tilapia mossambica as shown in Figure 1.The chill point of Tilapia mossambica is-1.2 ℃ of left and right, therefore can think after the reserve temperature of Tilapia Fillet is down to freezing point continuous decrease time, starts crystallization in fillet cell again, destroys eucaryotic cell structure.According to the freezing point of the Tilapia Fillet recording, experiment is that Tilapia Fillet is down to behind ice temperate zone herein, and fillet temperature is positioned within the scope of (0.5 ± 0.3) ℃ and remains unchanged.
Determining of 2.2 fish lethal temperatures
Through check, the lethal temperature of determining Tilapia mossambica is 12 ℃.
The variation of 2.3 fillet temperature
Before Tilapia mossambica fillet, 30h temperature lowering curve as shown in Figure 2.Presentation of results: the temperature lowering curve of mode I is evenly downward trend continuously, and mode II temperature lowering curve declines in gradient, 2 kinds of methods are to arrive ice temperate zone in 17h left and right substantially.Mode III is directly to put under ice temperature environment, and decrease speed is very fast, is just down to ice temperate zone in 3h left and right.
The variation of 2.4 K values
Fig. 3 is the high-efficient liquid phase chromatogram that contains 6 kinds of standard items ATP, ADP, AMP, IMP, HxR, Hx, and ATP and catabolite thereof just can effectively be separated in 18min under this chromatographic condition, and favorable reproducibility.Index of fish freshness K value be reflection aquatic products initial stage fresh target change and the relevant biochemical indicator of local flavor (Wu Chengye, Ye Mei, Wang Qin etc. several fresh-water fishes are fresh target change research in during frozen storage).The less expression freshness of K value is better, and K value more freshness is poorer.Many scholars are by the relation between the aquatic products research K values such as sardine, scad, conger pile, large yellow croaker and freshness, and K value is a kind of index of the generally acknowledged early stage freshness of evaluation fish, kills the K value of fish below 10%.As the K value of eating fillet raw, in one-level freshness below 20%, 20% ~ 40% is secondary freshness, and 60% ~ 80% is incipient spoilage fish.In ATP degradation process, produce 5 kinds of compounds such as ADP, AMP, IMP, HxR and Hx, the wherein ratio of HxR+Hx and ATP and catabolite summation thereof, is K value, can be represented by the formula:
By calculating, under 3 kinds of cold inductive conditions, the variation of Tilapia mossambica K value as shown in Figure 4.As can be seen from Figure 4,, in 10h, it is little that the K value of 3 kinds of cold induction modes changes difference.10～53h, the K value of 3 kinds of modes is all in rising trend, and mode III is considerably slower than another 2 kinds.After 53h, different cold induction modes demonstrate significant difference to the impact of K value, and mode I is at 53h, and K value is 16.2%, still can be used as raw fish edible, and in the time of 65h, K value rises to 25.6%, has exceeded 20% of one-level freshness; Mode II is at 65h, and K value is 18.3%, until when 90h, K value has just reached 20%.Mode III is in the time of 65h, and K value is 11.7%, and in the time of 90h, K value is only 13.8%, much smaller than first two induction mode, until when 126h, K value is 22.2%.Different cold induction modes have larger impact to K value, and possible reason is, in same time, oppressing residing mean temperature is that mode I is greater than mode II, and mode II is greater than mode III, has affected the enzymatic activity of ATP and relationship compound thereof, causes K value variation difference.In whole experimentation, 3 kinds of cooling methods all remain on relatively high freshness level, and when 138h, K value is all lower than 40%.
2.5 nucleotides IMP and ATP analyze
Inosinicacid (IMP) is atriphos (ATP) degraded, is the main taste compound of ucleotides, as novel food additives, has been widely used in foodstuff flavouring.IMP and sodium glutamate are mixed with synergy very significantly, experiment shows, if both mix in certain proportion, its delicate flavour can be greater than own delicate flavour (Li Huifang separately, Chen Guohong, Wu Xinsheng etc. animal muscle inosinicacid progress [J]. zoology and animal medicine, 1999,16 (4): 6-7.).IMP is the extremely strong flavour enhancer (Qi Xiaoyu of a kind of delicate flavour, Li Yan, Zhou Peigen. during Macrobrachium nipponensis iced storage, ATP catabolite changes and freshness evaluation), along with the decomposition of ATP, IMP is the trend of rising, in general within 1 ~ 2d, reach maximum concentration, and maintain a period of time, this is consistent with experimental result.The content of three kinds of cold induction mode ATP just presents the trend of fast-descending at front 20h as can be seen from Figure 5, and fall reaches respectively 92%, 93%, 93%.The fast degradation of ATP is relevant with highly active ATP enzyme, (Watabe S, Ushio H, Iwamoto M, the et al. Temperature-dependency of rigor-mort is of fish muscle:myofibrillar Mg such as Wataba
2+-ATPase activity and Ca
2+uptake by sarcoplasmic reticulum [J]. J Food Sci, 1989,54:1107-1115.) think, the calcium absorbability of sacroplasmine reticulum declines at low temperatures, and in muscle fibril, calcium concentration increases, calcium ion activated muscle fibril Mg
2+-ATP enzyme, has accelerated the degraded of ATP.IMP content is in rising trend and reach peak value before 20h, after this on a declining curve.At 17h place, the IMP content maximum of the mode I that records, II, III is respectively 166.43mg/100g, 165.62mg/100g, 167.75mg/100g, than distinguishing many 30%, 28%, 5% at 10h place, than 39h place respectively many 27%, 32%, 7.8%, 3 kind of mode IMP at 17 ~ 27h all in higher level.After IMP content 50h, have obvious difference, the IMP of mode I declines obvious, and mode II is taken second place, and it is the slowest that mode III declines.Experimental result illustrates in the later stage of cold induction, and different cooling methods affects IMP decomposition rate.Mode I is obviously accelerated in cold induction later stage IMP degraded than mode II, and the IMP degraded of mode III is slower, to the still content of IMP when just slaughtering of the IMP of 138h place content.
The analysis of 2.6 different cold induction mode Main Free Amino Acids
Under the different induction mode of table 1, Tilapia Fillet flavour free amino acid changes
Note: I-temperature control continuous cooling mode, II-temperature control ladder cooling method, III-direct ice temperature cooling method, TFAA-free amino acid total amount (comprising 17 seed amino acid asparatates, threonine, serine, glutamic acid, glycine, alanine, cystine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, histidine, arginine, proline).
Table 1 has been listed 8 kinds of flavour free aminoacid contents of Tilapia Fillet and the free amino acid total amount situation of change at front 65h.Because the IMP of 3 kinds of cold induction modes after 65h obviously declines, in table, do not list variation thereafter.
The TFAA of mode I, II, III, the 0h comparison after slaughtering with fish, at 17h place, increases and is respectively 35.96%, 21.86%, 20.03%; Increased by 38.61%, 26.03%, 21.68% at 27h place, increased by 49.61%, 29.7%, 25.2% at 65h place, 3 kinds of method TFAA content are in rising trend generally, and mode I TFAA growth trend is obvious, higher than mode II, III.In the flesh of fish, delicious taste is mainly to be determined by amino acid such as asparatate, glycine, glutamic acid, alanine and arginine, result shows that these four kinds of fillet Mid-Heaven Gate winter propylhomoserin, glycine, glutamic acid, the alanine of 0h are delicate flavour free amino acid and have just accounted for 58.04% of TFAA, and these compositions are apparent on the impact of flesh of fish local flavor.
Tilapia mossambica be mainly taste free amino acid in the content of glycine and alanine relatively high, glycine is the taste components of seafood products, has synergism with other delicate flavour material glutamic acid, inosinicacid etc.Mode I, II, III, 3 kinds of cold abductive approach, the 0h comparison after slaughtering with fish, at 17h place, the content of glycine has increased by 47.07%, 13.81%, 7.38%; At 27h place, increase by 51.58%, 26.25%, 19.54%; Increase by 76.16%, 27.24%, 26.44% at 65h place.In mode I, increase obviously, exceeded threshold value 130mg/100g at 17h, mode II, III increase slower after 27h.Alanine is slightly bitter sweet taste amino acid, also can draw the delicate flavour composition of fish and shellfish.In mode I, alanine has reached 14.04% in 27h recruitment, and mode II has only increased by 4.8%, and mode III does not increase variation substantially, and alanine content in whole process fluctuates at 20mg/100g, and variation tendency is milder.The threshold value of the glutamic acid of the flesh of fish is many below 0.03%, but with the flesh of fish in the IMP that accumulates produce synergism, even if content still can produce delicate flavour below threshold value.I, II, III, the content of glutamic acid in 3 kinds of methods is the trend of growth, has increased respectively 22%, 26%, 3% at 27h than 0h, and front 2 kinds of method recruitments are basically identical, and content is only second to the threshold value 5mg/100g of glutamic acid.Histidine be the speciality that causes some marine product " meat perfume (or spice) " (Shen Yuexin. aquatic food is learned [M]. the .2001:31-35. of Chinese agriculture publishing house), be respectively 17.17mg/100g, 16.70mg/100g, 15.96mg/100g at the histidine content at 27h place, substantially approach threshold value 20mg/100g, increased respectively 86.26%, 81.21%, 73.14% than 0h.
Arginine is a kind of bitter taste amino acid, content is less, in aquatic products, be not bitter taste, increase and put forward fresh effect on the contrary, the effect of continuation, complexity and dense sense to mouthfeel can not be ignored (Li Huifang, Chen Guohong, Wu Xinsheng etc. animal muscle inosinicacid progress [J]. zoology and animal medicine, 1999,16 (4): 6-7.).The amino acid methionine of bitter taste, fluctuate at 10mg/100g, variation tendency is not obvious and lower than threshold value 30mg/100g, it is the indispensable taste composition of sea urchin peculiar flavour, research find micro-methionine be improved the effect that MSG is sense of taste (Deng Jiechun, Wang Xichang, Liu Yuan. fugu obscurus is studied [J] with Fugu rubripes flavour component difference. food industry science and technology, 2010, (3): 106-108.).Isoleucine is also the amino acid that is bitter taste, the lower threshold value 90mg/100g well below them of content in whole temperature-fall period, and have the sign of minimizing; The asparatate with delicate flavour also has slightly minimizing, but because its content is low, limited on overall flavour impact.
3 conclusions
By adopting 3 kinds of different cold induction modes to carry out cold induction experimental study to Tilapia Fillet, draw the following conclusions:
(1) the K value of cold induction mode I, II, III, respectively in 60h, 90h, 120h, does not all exceed 20%, meets the food sanitation standard of fresh fillet; Different cold induction modes have time-lag effect to the impact of K value, and after 53h, the K value of mode I increases very fast, show that freshness declines very fast.
(2) nucleotides of cold induction mode III (IMP) increases soon, and later stage degradation is slow, and the Tilapia Fillet after slaughtering is down to ice temperate zone fast, can significantly delay the decline of IMP, is conducive to the reservation to its corresponding flavour.
(3) in same time, cold induction mode I can make Tilapia Fillet Free Amino Acids total amount obviously increase, and particularly glycine content obviously increases, and exceedes threshold value 130mg/100g in the time of 17h.
(4) consider K value, nucleotides (IMP) and the mainly variation of flavour free amino acid under different condition, the Tilapia Fillet of mode I induction, the most delicious during 17 ~ 27h.
1 materials and methods
1.1 materials and equipment
Tilapia mossambica: in August, 2012 Tilapia mossambica, purchase in food market, Lingang New City Gu Zong road, Shanghai City, body weight 500 ~ 600g/ tail, buy after oxygenation transport rapidly laboratory back.Live fish is placed in to keep-alive case and stores up 1 ~ 2h, avoid its struggle, then live fish is knocked lethal after, sheet becomes fillet, is distributed into freshness protection package and is placed in cold environment; Atriphos (ATP), adenosine diphosphate (ADP) (ADP), inosinicacid (IMP), hypoxanthine (Hx): Sigma company; Adenosine monophosphate (AMP): Japanese TCI company; Inosine (HxR): German Dr.Ehrenstorfer company; Dipotassium hydrogen phosphate, potassium dihydrogen phosphate: Town in Shanghai spectrum scientific instrument company, chromatographically pure; Ultra-pure water, perchloric acid (PCA), potassium hydroxide, NaOH, phosphoric acid, sulfosalicylic acid: Chemical Reagent Co., Ltd., Sinopharm Group, analyzes pure; Methyl alcohol: Chemical Reagent Co., Ltd., Sinopharm Group, chromatographically pure.
LC-2010CHT high performance liquid chromatograph, AUW320 electronic balance: Japanese Shimadzu company; L-8800 amino acid fully-automatic analyzer: HITACHI company; Agilent-34972A temperature sensor: Agilent company; BPS-250CB(-20 ~ 100 ℃) climatic chamber, DHG-9053A electric drying oven with forced convection: Shanghai Yiheng Scientific Instruments Co., Ltd; The full-automatic snowflake ice machine of IMS-50: Changshu Xue Ke Co., Ltd; SB25-12DT ultrasonic machine: the new sesame biotechnology in Ningbo; FA25 homogenizer: Fluko company; PHS-3C type pH meter: Shanghai Precision Scientific Apparatus Co., Ltd; H2050R refrigerated centrifuge: Changsha Xiang Yi Co., Ltd; GM-0.33A vacuum diaphragm pump and solvent filter: Tianjin Jin Teng experimental facilities Co., Ltd; Haier BCD-216SCM refrigerator: Qingdao company of Haier.The equipment of experiment use all adopts ultrasonic machine to clean 20min, repeatedly rinses with distilled water, then uses ultra-pure water rinse 2 ~ 3 times, and drying machine drying is for subsequent use.
1.2 experimental technique
1.2.1 freezing test
Along vertebra, fish being cutd open is two, and thermocouple is inserted to about 0.5cm place fixing under fish body surface, puts into the cold storage chamber of-20 ℃, and temperature acquisition is spaced apart 10s and gathers a secondary data, and experiment finishes rear drafting freezing curve and draws Tilapia mossambica freezing point.
1.2.2 the setting of the cold induction mode cooling process of different cold induction initial temperatures
5 kinds of different initial temperatures, are respectively 25,20,15,10,5 ℃, all adopt temperature control cooling.Cooling process is in table 2.
The setting of table 2 cooling process
With same rate of temperature fall cooling, allow the flesh of fish adapt to slowly cold environment, until be down to ice temperate zone, and maintain ice temperature condition a period of time.
1.2.3 the detection of ATP and relationship compound thereof
With reference to Yokoyama(Yokoyama Y, Sakaguchi M, Kawai F, et al. Change in concentration of ATP-related compounds in varioustissues of oyster during ice storage[J]. Nippon Suisan Gakkaishi, 1992,58 (11): 2125-2136.) method, slightly changes.The extraction of fillet ATP and relationship compound: get fresh fillet and the sample after cold treatment, get 5g after chopping fast and put into centrifuge tube, add perchloric acid (PCA) the solution making beating 2min of the 10mL 10% of precooling.After homogenate, with 10000r/min refrigerated centrifuge 15min, get supernatant.5% PCA washing of precooling for precipitation, centrifuging and taking supernatant, repeats once.Merge supernatant, first regulate pH value with the KOH solution of 10mol/L, in the time approaching required pH value, the KOH solution essence adjust pH of using 1mol/L instead is 6.5, after leaving standstill 30min, supernatant is transferred in 50mL volumetric flask, and use ultra-pure water constant volume, shake up, with to be determined after 0.45 μ m micro porous filtration membrane filtration etc.Whole process all operates under 0 ~ 4 ℃ of condition.
High performance liquid chromatography (HPLC) detects chromatographic condition: the Inertsil ODS-SP C of GL Sciences company
18(4.6mm × 250mm, m) liquid-phase chromatographic column of 5 μ; Guard column post core Inertsil ODS-SP (4mm × 10mm, 5 μ are m); Mobile phase: A is 0.05mol/L potassium dihydrogen phosphate and dipotassium hydrogen phosphate (1:1) solution, and being adjusted to pH with phosphoric acid is that 6.5, B is methanol solution; Isocratic elution; Flow velocity: 1mL/min; Column temperature: 28 ℃; Sample size: 10 μ L; Check wavelength: 254nm.
1.2.4 the detection of free amino acid
With reference to Deng Jiechun (Deng Jiechun, Wang Xichang, Liu Yuan. fugu obscurus and Fugu rubripes flavour component difference research [J]. food industry science and technology, 2010,31 (3): 106-108.) free amino acid assay method, slightly change.The extraction of free amino acid: take respectively flesh of fish 2g, add 10mL 5%(w/v) sulfosalicylic acid, precipitation 2h gets off protein precipitation, draw the centrifugal 15min of 6mL supernatant 10000r/min refrigerated centrifuge, get 3mL supernatant, with a certain amount of NaOH solution adjusting pH to 2.0 left and right, be settled to 12mL, measure with being filled to upper machine in sample disc after 0.45 μ m micro porous filtration.
Analysis condition: L-8800 type automatic amino acid analyzer, sample analysis cycle 53min.Chromatographic column (4.6mm × 150mm, 7 μ are m); Column temperature: 50 ℃; Passage 1 flow velocity: 0.4mL/min, passage 2 flow velocitys: 0.35mL/min.Mobile phase: it is the buffer solution of 4% ninhydrin that pH value is respectively 3.3,3.2,4.0,4.9 citric acid and natrium citricum cocktail buffer and concentration.
2 results and discussion
2.1 freezing test
The freezing curve of Tilapia mossambica as shown in Figure 1.The chill point of Tilapia mossambica is-1.2 ℃ of left and right, therefore can think after the reserve temperature of Tilapia Fillet is down to freezing point continuous decrease time, starts crystallization in fillet cell again, destroys eucaryotic cell structure.According to the freezing point of the Tilapia Fillet recording, experiment is that Tilapia Fillet is down to behind ice temperate zone herein, and fillet temperature is positioned within the scope of (0.5 ± 0.3) ℃ and remains unchanged.
The variation of 2.2 fillet temperature
Before Tilapia mossambica fillet, 35h temperature lowering curve as shown in Figure 6.Result shows: mode I, II, III, IV, V temperature lowering curve are all continuous cooling trend, and respectively 27,22,17,12,7h left and right arrives ice temperate zone, and maintains ice temperate zone a period of time, keeps Tilapia mossambica fillet temperature in (0.5 ± 0.3) ℃.
The variation of 2.3 index of fish freshness K values
By calculating, under the different cold induction mode of I, II, III, IV, 5 kinds of initial temperatures of V, the variation of Tilapia mossambica K value as shown in Figure 7.As seen from Figure 7, the impact of the cold induction mode of 5 kinds of different initial temperatures on Tilapia mossambica fillet K value, K value is in temperature-fall period, elongated with storage time, all in rising trend, and the K value of fresh fillet is 2.27%.Mode I and II K value rise very fast, and mode III is taken second place, and mode IV and V increases slowly, and are mild ascendant trend.After mode I induction 27h, K value is just to 20%, also within the scope of one-level freshness; After 28 ~ 70h, K value exceedes 40%, and fillet are in the secondary freshness stage; Be 46.1% in 171h K value, also do not arrive putrefaction stage.Mode II, at 30h place, exceedes one-level index of fish freshness 20%, all exceedes 30%, and approach secondary index of fish freshness at 142h in the K of 70 ~ 142h place value.Mode III is at 50h still within the scope of one-level index of fish freshness, until 161h, K value 36.7%, within the scope of secondary index of fish freshness.Mode IV and mode V, arrive one-level index of fish freshness at 108h and 127h respectively, do not exceed 30% to 150h, maintains higher freshness level.
2.4 nucleotides IMP and ATP analyze
Inosinicacid (IMP), by atriphos (ATP) degraded, is the main taste compound of ucleotides, has been widely used in foodstuff flavouring.IMP is the extremely strong flavour enhancer of a kind of delicate flavour, and along with the decomposition of ATP, IMP is the trend of rising, in general within 1 ~ 2d, reaches maximum concentration, and maintains a period of time, and this is consistent with experimental result.The fast degradation of ATP is relevant with highly active ATP enzyme, (the Watabe S such as Wataba, Ushio H, Iwamoto M, et al. Temperature-dependency of rigor-mort is of fish muscle:myofibrillar Mg2+-ATPase activity and Ca2+ uptake by sarcoplasmic reticulum[J]. J Food Sci, 1989,54:1107-1115.) think, the calcium absorbability of sacroplasmine reticulum declines at low temperatures, in muscle fibril, calcium concentration increases, calcium ion activated muscle fibril Mg
2+-ATP enzyme, has accelerated the degraded of ATP.Under the cold inductive condition of different initial temperatures the variation of Tilapia mossambica IMP and ATP as shown in Figure 8, as shown in Figure 8, within 40h, all there is peak value in IMP, with friction speed degraded, the degradation speed of mode I and mode II is very fast after 40h, at 50h place when just slaughtering IMP value.Mode III, IV be IMP value 2.62 μ mol/g when just slaughtering, and the time is respectively 89,150h.The IMP content of mode V is always in higher level, and experiment finishes still higher than the level of just having slaughtered.
The analysis of 2.5 free amino acids
Table 3 Tilapia Fillet flavour free amino acid composition (mg/100g)
Note: I: 20 ℃ of cold induction modes that are initial temperature, II: 15 ℃ of cold induction modes that are initial temperature, III: 10 ℃ of cold induction modes that are initial temperature, IV: 15 ℃ of cold induction modes that are initial temperature, V: 5 ℃ of cold induction modes that are initial temperature; TFAA comprises 17 seed amino acids (asparatate, threonine, serine, glutamic acid, glycine, alanine, cystine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, histidine, arginine, proline).
Table 3 has been listed the situation of change of 6 kinds of flavour free amino acids of Tilapia mossambica and free amino acid total amount.With regard to TFAA, mode I, the equal growth trend of II are mild, and mode III, IV have obvious increase, and mode V growth rate is the slowest.0h comparison after slaughtering with fish, at 27h place, has increased respectively 10.81%, 10.31%, 21.46%, 14.87%, 4.97%; At 75h place, increase respectively 15.55%, 13.31%, 32.01%, 15.01%, 8.71%; At 123h place, increase respectively 16.43%, 19.19%, 32.63%, 25.36%, 14.91%.The cold induction mode TFAA content of above 5 kinds of different initial temperatures, all in rising trend, mode III, IV growth trend are obvious, and mode III is greater than mode V, all higher than mode I, II and V.The delicious taste of the flesh of fish is mainly to be determined by amino acid such as glycine, glutamic acid, alanine, leucine, isoleucine, histidines, experimental result shows, in the Tilapia Fillet of just having slaughtered, these 4 kinds main taste free amino acids that are of asparatate, glutamic acid, glycine and alanine have just accounted for 57.28% of the TFAA recording, and these compositions are very obvious on the impact of flesh of fish flavour.
Fish free amino acid content compared with shellfish is lower, but some free amino acid can be present in the flesh of fish with enough high concentration, to the local flavor generation effect of fish.In being in taste free amino acid of Tilapia mossambica fillet, glycine and alanine content are relatively high, and wherein glycine is the main taste compound of seafood products, have synergism with delicate flavour material inosinicacid, glutamic acid etc.The cold abductive approach of 5 kinds of different initial temperatures, at 27h place, relatively, the glycine content of mode III is the highest, approach the threshold value 130mg/100g of glycine, increased by 34.18% than former state, mode I, II, IV have increased respectively 20.98%, 18.65%, 28.10%, mode V growth trend minimum.Fish and shellfish all contains glutamic acid, the threshold value of the glutamic acid in the flesh of fish below 0.03%, but produce synergism with the inosinicacid in muscle after death, so even content in threshold value still can produce delicate flavour
[17].5 kinds of cooling methods with glutamic acid, substantially all exceed threshold value 5mg/100g at 27h place, are respectively 16.04%, 20.37%, 23.12%, 23.74%, 2.89% than the recruitment of former state.Be respectively 5.74%, 30.30%, 37.07%, 18.65%, 10.53% in the recruitment at 75h place, mode III glutamic acid growth trend is relatively obvious.Alanine is sweet taste amino acid, and the delicate flavour of the flesh of fish is had to contribution.In 5 kinds of modes, the content of alanine fluctuates at 21mg/100g, changes milder.Histidine is the speciality that causes some marine product meat perfume (or spice), and mode I, II, III increase are more remarkable, have increased by 69.41%, 44.50%, 95.22% at 27h place than 0h place, and mode IV, V do not increase variation substantially.
Leucine and isoleucine are the amino acid that is bitter taste, and result of study finds that the content of leucine and methionine is far below their threshold value 190mg/100g and 90mg/100g, and in temperature-fall period, has minimizing trend.In addition, arginine content and methionine are also bitter taste amino acid, and continuation and the complexity of arginine to mouthfeel has the effect can not be ignored, and methionine is the indispensable taste composition of sea urchin local flavor.
3 conclusions
3.1 cold induction mode I, II, III, IV, V, on Tilapia mossambica freshness, impact is different.Initial inducing temperature is lower, and the growth of K value is slower, and freshness is better, otherwise the growth of K value is very fast, and freshness declines.
3.2 cold induction mode I, II, III, IV, V, all in 40h, there is peak value in IMP content, in this while, ATP degrades rapidly.
3.3 cold induction mode I, II, III, IV, V, free amino acid total amount all increases to some extent, different induction mode amplification degree differences, research finds that cold induction initial temperature is 15 ℃, free amino acid recruitment is more remarkable.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, do not departing under the prerequisite of the inventive method; can also make some improvement and supplement, these improvement and the supplementary protection scope of the present invention that also should be considered as.
Claims (6)
1. the cold induction of ice temperature keeps fish freshness and increases a method of oppressing flavour, it is characterized in that, it comprises the following steps:
A) flesh of fish freezing test;
B) determine the lethal temperature of fish;
C) processing before the cold induction of live fish: live fish more than temporary foster 1h, is slaughtered fish in the water higher than 3～5 ℃ of lethal temperatures in the environment higher than 3～5 ℃ of lethal temperatures, gilled, whole piece, segmentation or section;
D) cold induction: flesh of fish temperature is down to behind its ice temperate zone, in ice temperate zone storage a period of time, wherein, described its ice temperate zone that flesh of fish temperature is down to refers to that the flesh of fish is put into cold environment carries out the induction of temperature control continuous cooling or temperature control ladder cooling induction, oppress the initial temperature of cold induction higher than 3～5 ℃ of lethal temperatures, the finishing temperature of oppressing cold induction is its ice temperate zone, the program of described temperature control continuous cooling induction is: per hourly between initial temperature → 1 ℃ fall 1 ℃, 1 ℃ → the flesh of fish is per hour between freezing point falls 0.5 ℃, the program of described temperature control ladder cooling induction is: be directly down to 10 ℃ and keep temperature-resistant from initial temperature, when starting frigorimeter, initial temperature is directly down to again 4 ℃ and keep temperature-resistant 5h, during since 10 ℃ of frigorimeters, after 10h, be directly down to again flesh of fish freezing point.
2. method according to claim 1, it is characterized in that, its concrete way of flesh of fish freezing test described in step a) is: fish is slaughtered, gill, clean up, by thermoelectricity occasionally thermal resistance equitemperature sensor insert under fish body surface about 0.5cm place and fix, put into the cold storage chamber of-20 ℃, temperature acquisition is spaced apart 10s and gathers a secondary data, and experiment finishes rear drafting freezing curve and draws the freezing point of the flesh of fish.
3. method according to claim 1, it is characterized in that, its concrete way of the lethal temperature of definite fish described in step b) is: live fish is placed in ecological tank and is supported temporarily, with the speed of 1 ℃/24h to water for cooling, death time and corresponding water temperature situation are once recorded in every 2h manual observation, the death standard of check experiment fish is, the gill cover stop agitating and acupuncture reactionless, after experiment fish is placed in to natural temperature, in 5 minutes, do not reactivate, 3 repeated experiments, chooses the maximum temperature while there is dead fish, as the lethal temperature of fish.
4. a method of determining the cold induction mode of the best ice temperature of the flesh of fish, is characterized in that, it comprises the following steps:
A) flesh of fish freezing test;
B) determine the lethal temperature of fish;
C) processing before the cold induction of live fish: live fish more than temporary foster 1h, is slaughtered fish in the water higher than 3～5 ℃ of lethal temperatures in the environment higher than 3～5 ℃ of lethal temperatures, gilled, whole piece, segmentation or section;
D) cold induction: flesh of fish temperature is down to behind its ice temperate zone, in ice temperate zone storage a period of time, wherein, described its ice temperate zone that flesh of fish temperature is down to refers to that the flesh of fish is put into cold environment carries out the induction of temperature control continuous cooling or temperature control ladder cooling induction, oppress the initial temperature of cold induction higher than 3～5 ℃ of lethal temperatures, the finishing temperature of oppressing cold induction is its ice temperate zone, the program of described temperature control continuous cooling induction is: per hourly between initial temperature → 1 ℃ fall 1 ℃, 1 ℃ → the flesh of fish is per hour between freezing point falls 0.5 ℃, the program of described temperature control ladder cooling induction is: be directly down to 10 ℃ and keep temperature-resistant from initial temperature, when starting frigorimeter, initial temperature is directly down to again 4 ℃ and keep temperature-resistant 5h, during since 10 ℃ of frigorimeters, after 10h, be directly down to again flesh of fish freezing point,
E) measure index of fish freshness K value and flavour composition, the flesh of fish of the cold induction mode of difference is carried out to the evaluation of freshness and flavour, determine best cold induction mode.
5. method according to claim 4, it is characterized in that, its concrete way of flesh of fish freezing test described in step a) is: fish is slaughtered, gill, clean up, by occasionally thermal resistance equitemperature sensor of thermoelectricity, insert about 0.5cm place fixing under fish body surface, put into the cold storage chamber of-20 ℃, temperature acquisition is spaced apart 10s and gathers a secondary data, and experiment finishes rear drafting freezing curve and draws the freezing point of the flesh of fish.
6. method according to claim 4, it is characterized in that, its concrete way of the lethal temperature of definite fish described in step b) is: live fish is placed in ecological tank and is supported temporarily, with the speed of 1 ℃/24h to water for cooling, death time and corresponding water temperature situation are once recorded in every 2h manual observation, the death standard of check experiment fish is, the gill cover stop agitating and acupuncture reactionless, after experiment fish is placed in to natural temperature, in 5 minutes, do not reactivate, 3 repeated experiments, chooses the maximum temperature while there is dead fish, as the lethal temperature of fish.
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