CN110982854A - Preparation method of salmon viscera fish oil - Google Patents
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Abstract
The invention discloses a preparation method of salmon viscera fish oil, which comprises the steps of raw material pretreatment, first-stage enzymolysis and second-stage enzymolysis; the first-stage enzymolysis is mainly lipase enzymolysis, meanwhile, the autolytic enzyme enzymolysis process is assisted to be enhanced, and calcium chloride is added in the process; the second-stage enzymolysis is protease enzymolysis. The method can obviously shorten the extraction time of the fish oil, obviously improve the extraction rate of the fish oil, simultaneously the prepared crude visceral fish oil has orange, bright and clear color, no fishy smell and no peculiar smell, obviously improves the quality, effectively reduces the rancidity degree in the extraction process of the fish oil, improves the acceptability of the visceral fish oil, expands the application approach of the visceral fish oil and improves the processing applicability of the visceral fish oil.
Description
Technical Field
The invention belongs to the technical field of aquatic product processing, and particularly relates to a preparation method of salmon visceral fish oil.
Background
The salmon as a rare product in water has high nutritive value and contains rich high-quality protein, vitamin D and n-3 unsaturated fatty acid, wherein the content of the unsaturated fatty acid in the fish oil reaches more than 60 percent, and clinical researches prove that the n-3 polyunsaturated fatty acid has physiological effects of promoting brain development, reducing the risk of cardiovascular diseases, resisting inflammation, resisting tumors and the like.
The consumption of salmon is gradually increased in China, large-scale culture is formed in China, because a large amount of visceral leftovers generated in the process of killing the salmon contain rich grease and can be used as a production raw material of high-quality fish oil, however, in view of the fact that salmon is large in size and has viscera such as fish liver, fish intestine and the like naturally larger than other common fishes, a large amount of dirty blood can be remained in the liver after killing, particularly, the fish liver contains a large amount of grease but is dark in color, the appearance and color of the fish oil can be seriously influenced, a large amount of excrement, microorganisms and the like can also be remained in a thick intestinal tract, if the grease extraction process cannot be effectively controlled, the extracted visceral crude fish oil is turbid, the fishy smell is obvious, the color is not clear and transparent, the quality of the fish oil is seriously influenced, the application range of the fish oil is limited, and the difficulty of subsequent fish oil refining is increased. The salmon viscera has strong autolysis, and if the fish liver contains rich endogenous enzymes (mainly endogenous protease), the extraction effect of the fish oil is influenced, the resource waste is caused, and the environmental pollution problem is caused. Therefore, the extraction of visceral fish oil with excellent sensory properties and good flavor aiming at the unique characteristics of the salmon viscera is an urgent technical problem to be solved.
At present, the crude visceral fish oil is extracted mainly by an enzymolysis technology, although the extraction condition is mild and the damage to active ingredients of the fish oil is small, if the relative speed of the precipitation of oil and the emulsification of the oil is not controlled well, the extraction rate of the oil is reduced, and pigments, fishy smell and small molecular substances with bad flavor in visceral tissues are embedded by the oil due to the emulsification of the oil, so that the quality of the fish oil is influenced.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides the preparation method of the salmon viscera fish oil, the method can obviously shorten the extraction time of the fish oil and obviously improve the extraction rate of the fish oil, and meanwhile, the prepared crude viscera fish oil has orange yellow, bright and clear color, basically no fishy smell and no peculiar smell, obviously improves the quality, improves the acceptability of the viscera fish oil, expands the application approach of the viscera fish oil and improves the processing applicability of the viscera fish oil. In addition, the method can effectively reduce the rancidity degree in the extraction process of the fish oil, and has unexpected technical effects.
The specific technical scheme is as follows:
a method for preparing salmon viscera fish oil comprises pretreating raw materials, performing first-stage enzymolysis, and performing second-stage enzymolysis;
the first-stage enzymolysis is mainly lipase enzymolysis, meanwhile, the autolytic enzyme enzymolysis process is assisted to be enhanced, and calcium chloride is added in the process.
The second-stage enzymolysis is protease enzymolysis.
The invention adopts the technical means of sectional enzymolysis to extract crude fish oil from the internal organs of the salmon, breaks through the inherent mode of extracting the fish oil by the conventional enzyme method, uses lipase to combine with autolytic enzyme in the internal organs of the salmon in the first-stage enzymolysis, kills the enzyme after the first-stage enzymolysis is carried out for a period of time, separates the upper-layer fish oil, uses protease to carry out enzymolysis on a tissue system independently, and separates the upper-layer fish oil again. The whole process adopts a linkage, quick and thorough extraction mode, and effectively solves the technical problems of incomplete extraction and poor quality of the visceral fish oil rich in grease.
Enzymatic extraction of fish oil in the art typically uses only proteases to disintegrate the proteinaceous tissue to release the fish oil. However, when the method is applied to the extraction of the internal organs of the salmon, the reaction condition is difficult to control, and serious emulsification is generated after enzymolysis for a certain time, so that a large amount of fish oil which is originally separated out is combined with protein emulsion and protein liquid in a short time, and the ideal yield of the fish oil is difficult to achieve. If the use amount of enzyme is increased after emulsification, the enzymolysis is continued for a long time, most protein tissues are enzymolyzed into small peptides, an emulsification system is damaged, and emulsified fish oil can be partially separated out, but the whole color of the fish oil is dark and dark brown, which brings great difficulty for subsequent processing and is difficult to be used as a polyunsaturated fatty acid source for medicinal and high-end food. If the enzymolysis is stopped before emulsification occurs, the fish oil yield is very low.
Lipases are generally regarded in the art as being advantageous for emulsification and are therefore commonly used as emulsifiers; however, in the experiments for preparing the salmon visceral oil, the lipase used in the early stage of preparing the salmon visceral oil can be matched with autolytic enzyme contained in the viscera to accelerate the disintegration of the viscera tissues and quickly release a part of grease, and the part of grease is lighter and bright in color, bright orange and similar to the color of fish oil extracted by taking salmon meat as a raw material. It should be noted that it is necessary to deactivate enzyme and stop reaction before the emulsifying speed exceeds the oil separating speed, so as to ensure high extraction rate and good organoleptic properties. And (3) separating the fish oil after the lipase reaction is terminated, adding protease to carry out enzymolysis on the residual tissue fluid, promoting the sufficient release of grease in the tissue, and obtaining the two-stage enzymolysis precipitated fish oil.
Experiments prove that the method can obviously improve the yield of the salmon viscera fish oil in a relatively short extraction time, and is obviously superior to the traditional protease method. Meanwhile, the extraction time of the fish oil is shortened, so that the oxidation rancidity degree of the fish oil in the extraction process can be effectively reduced, the embedding and dissolving effects of the oil on pigments, impurities and the like in a system are reduced, and the sensory properties and quality of the fish oil are obviously improved.
Preferably, the working conditions of the first stage of enzymolysis are as follows:
mixing the pretreated visceral tissue with water according to a mass ratio of 1: 0.5-1: 2, adding 1.0-1.3 wt% of calcium chloride based on the mass of the added water, adding 100-160 u/g of lipase based on the mass of a reaction system, carrying out enzymolysis at 40-55 ℃, carrying out enzyme deactivation after carrying out enzymolysis for 1-2 h, cooling to below 25 ℃, and carrying out centrifugal separation to obtain the upper-layer crude fish oil.
The enzyme deactivation method can adopt water bath heating at 90-95 ℃ for 5-10 min.
The first stage of enzymolysis is the combined action of lipase and autolytic enzyme of internal organs. The limitation of the enzymolysis time is important, and the emulsification is prevented from exceeding the oil precipitation speed and generating obvious emulsification. Inactivation of the enzyme after enzymatic hydrolysis is necessary, again to prevent the occurrence of excessive emulsification.
Stirring can be carried out in the first-stage enzymolysis process, the stirring in a certain rotating speed range is favorable for the reaction, but the too fast rotating speed causes the emulsification to be accelerated. Preferably, the stirring speed is 60-100 r/min.
Preferably, the working conditions of the second stage enzymolysis are as follows:
adding protease into a system left after the first-stage enzymolysis crude fish oil is separated, wherein the addition amount of the protease is 1500-2500 u/g based on the mass of the remaining system, the enzymolysis temperature is 45-55 ℃, the pH value is 7.0-8.0, cooling to below 25 ℃ after enzymolysis is carried out for 1-2 hours, and carrying out centrifugal separation to obtain the upper-layer crude fish oil.
Further preferably, the protease is an animal protease.
Stirring can be carried out during the second-stage enzymolysis, and the stirring speed is preferably below 100 r/min. The stirring can be stopped after 20-30 min in the two-stage enzymolysis process.
Preferably, the working conditions of the raw material pretreatment are as follows: removing gallbladder, air bladder and feces.
In particular to a method for removing gall bladder and swimming bladder from fresh viscera or thawed viscera and extruding excrement in intestinal tracts.
The tissue can be broken into small pieces of 1-3 cm, but the tissue is not suitable to be twisted into erosive shape.
It should be noted that the washing of the internal organs is not desirable and leads to a reduction in yield.
The invention has the following beneficial effects:
the invention adopts the technical means of sectional enzymolysis to extract crude fish oil from the internal organs of the salmon, breaks through the inherent mode of extracting the fish oil by the conventional enzyme method, uses the combined action of lipase and autolytic enzyme in the internal organs of the salmon in the first-stage enzymolysis to accelerate the disintegration of the internal organs, simultaneously quickly releases most of grease, and then further destroys the internal organs by using protease. Promoting the sufficient release of grease in the tissues. By implementing the technical scheme, the extraction rate of the salmon viscera fish oil can be obviously improved, and meanwhile, the oxidation rancidity degree of the fish oil in the extraction process can be effectively reduced due to the shortened extraction time of the fish oil, the embedding effect of oil on pigments, impurities and the like in a system is reduced, and the sensory properties and quality of the fish oil are obviously improved.
Detailed Description
The principles and features of this invention are described below in conjunction with examples, which are set forth to illustrate, but are not to be construed to limit the scope of the invention.
The instruments, reagents, materials and the like used in the following examples are conventional instruments, reagents, materials and the like in the prior art and are commercially available in a normal manner unless otherwise specified. Unless otherwise specified, the experimental methods, detection methods, and the like described in the following examples are conventional experimental methods, detection methods, and the like in the prior art.
Lipase used in the following examples was purchased from Pengpo bioengineering, Inc., Nanning, with a viability unit of 1X 105u/g, animal protease purchased from Nanning Pompe bioengineering, Inc. with a viability unit of 1X 105u/g。
Example 1
A method for preparing salmon viscera fish oil comprises the following steps:
(1) pretreatment of raw materials: taking about 2Kg of fresh salmon viscera, removing gall bladder and swim bladder, extruding feces in the fish intestines, cutting into small pieces of 1-3 cm, and weighing 1.82Kg for later use.
(2) First-stage enzymolysis: mixing the visceral tissue obtained in the step (1) with water according to a mass ratio of 1:0.5, adding 1.0wt% of calcium chloride calculated by the mass of the added water, wherein the addition amount of lipase is 100u/g calculated by the mass of a reaction system, the enzymolysis temperature is 40 ℃, standing and carrying out enzymolysis for 2h under the condition of pH7.0, then heating at 90 ℃ for 10min for enzyme deactivation, cooling to below 25 ℃, centrifuging at 6000r/min for 15min, and collecting the upper-layer crude fish oil.
(3) Second-stage enzymolysis: and (3) carrying out proteolysis on the reaction system remained after the fish oil is collected in the step (2). Adding animal protease into the reaction system, wherein the addition amount is 1500u/g based on the rest reaction system, the enzymolysis temperature is 45 ℃, standing and performing enzymolysis for 1h under the condition of pH7.0, performing ice bath, cooling to below 25 ℃ within 20min, centrifuging at 6000r/min for 15min, and collecting the upper layer crude fish oil.
(4) And (3) combining the crude fish oil obtained in the step (2) and the step (3).
Example 2
A method for preparing salmon viscera fish oil comprises the following steps:
(1) pretreatment of raw materials: taking 1.5Kg of frozen salmon viscera, thawing, removing gallbladder and swim bladder, extruding feces in the fish intestines, weighing 1.3Kg for later use.
(2) First-stage enzymolysis: mixing the visceral tissue obtained in the step (1) with water according to a mass ratio of 1:2, adding 1.3wt% of calcium chloride in terms of the mass of the added water, wherein the addition amount of lipase is 160u/g in terms of the mass of a reaction system, the enzymolysis temperature is 55 ℃, the enzymolysis is carried out for 1h under the condition of pH8.5, and the stirring is continuously carried out in the enzymolysis process, wherein the stirring speed is 80 r/min; heating at 95 deg.C for 5min to inactivate enzyme after enzymolysis, cooling to below 25 deg.C, centrifuging at 6000r/min for 15min, and collecting upper layer crude fish oil.
(3) Second-stage enzymolysis: and (3) carrying out proteolysis on the reaction system remained after the fish oil is collected in the step (2). Adding animal protease into the reaction system at 2500 u/g based on the rest reaction system, allowing standing at 55 deg.C and pH8.0 for enzymolysis for 1.5h, cooling to below 25 deg.C in ice bath for 20min, centrifuging at 6000r/min for 15min, and collecting the upper layer crude fish oil.
(4) And (3) combining the crude fish oil obtained in the step (2) and the step (3).
Example 3
A method for preparing salmon viscera fish oil comprises the following steps:
(1) pretreatment of raw materials: taking 1.5Kg of frozen salmon viscera, thawing, removing gallbladder and swim bladder, extruding feces in the fish intestines, weighing 1.25Kg for later use.
(2) First-stage enzymolysis: mixing the visceral tissue obtained in the step (1) with water according to a mass ratio of 1:2, adding 1.2wt% of calcium chloride in terms of the mass of water, wherein the addition amount of lipase is 160u/g in terms of the mass of a reaction system, the enzymolysis temperature is 55 ℃, the enzymolysis is carried out for 1h under the condition of pH8.5, and the stirring is continuously carried out in the enzymolysis process, and the rotating speed of the stirring is 100 r/min; heating at 95 deg.C for 5min to inactivate enzyme after enzymolysis, cooling to below 25 deg.C within 20min, centrifuging at 6000r/min for 15min, and collecting upper layer crude fish oil.
(3) Second-stage enzymolysis: and (3) carrying out proteolysis on the reaction system remained after the fish oil is collected in the step (2). Adding animal protease into the reaction system, wherein the addition amount is 2000 u/g calculated by the rest reaction system, the enzymolysis temperature is 50 ℃, the enzymolysis is carried out for 2h under the condition of pH7.5, the stirring is continuously carried out for 20min before the enzymolysis, the stirring rotating speed is 60r/min, and the stirring is stopped after the enzymolysis is carried out for 20 min; and (4) after the enzymolysis is finished, performing ice bath, cooling to below 25 ℃ within 20min, centrifuging at 6000r/min for 15min, and collecting the upper-layer crude fish oil.
(4) And (3) combining the crude fish oil obtained in the step (2) and the step (3).
Example 4
A method for preparing salmon viscera fish oil comprises the following steps:
(1) pretreatment of raw materials: taking 2.3Kg of frozen salmon viscera, unfreezing, removing gallbladder and swim bladder, extruding feces in the fish intestines, and weighing 1.93Kg for later use.
(2) First-stage enzymolysis: mixing the visceral tissue obtained in the step (1) with water according to a mass ratio of 1:1, adding 1.3wt% of calcium chloride in terms of the mass of the added water, wherein the addition amount of lipase is 120u/g in terms of the mass of a reaction system, the enzymolysis temperature is 50 ℃, the enzymolysis is carried out for 1.5h under the condition of pH7.5, and the stirring is continuously carried out in the enzymolysis process, wherein the stirring speed is 60 r/min; heating at 95 deg.C for 5min to inactivate enzyme after enzymolysis, cooling to below 25 deg.C, centrifuging at 6000r/min for 15min, and collecting upper layer crude fish oil. At this point, the fish oil yield was calculated to be 23.8%.
(3) Second-stage enzymolysis: and (3) carrying out proteolysis on the reaction system remained after the fish oil is collected in the step (2). Adding animal protease into the reaction system, wherein the addition amount is 2000 u/g based on the rest reaction system, the enzymolysis temperature is 50 ℃, standing and enzymolysis are carried out for 1.5h under the condition of pH7.5, ice bath is carried out after the enzymolysis is finished, cooling is carried out to below 25 ℃ within 20min, 6000r/min is carried out for centrifugation for 15min, and the crude fish oil at the upper layer is collected.
(4) And (3) combining the crude fish oil obtained in the step (2) and the step (3).
Experimental example 1
The working conditions of experimental example 1 were as in example 4, with the only difference that no lipase was added in the first stage of the enzymatic hydrolysis.
Experimental example 2
The working conditions of experimental example 2 refer to example 4, and the only difference is that the first stage enzymolysis is not inactivated, and the second stage enzymolysis is carried out without adding animal protease, and lipase enzymolysis is still carried out.
Experimental example 3
The working conditions of the experimental example 3 refer to the working conditions of the example 4, and the difference is that the first stage enzymolysis is not carried out, the second stage enzymolysis is directly carried out, and the enzymolysis process is carried out by standing.
Experimental example 4
The working conditions of example 4 were as in example 3, except that the duration of the enzymolysis was 3 hours, which corresponds to the total duration of the enzymolysis in example 4.
Experimental example 5
Experimental example 5 the working conditions were as in Experimental example 4, except that the stirring was continued during the enzymatic hydrolysis at a speed of 100 r/min. The method of experimental example 5 corresponds to the enzymatic preparation of fish oil commonly used in the prior art. (except for the rotation speed, the technical scheme of the experimental example 5 is basically the same as that of the prior art, and the rotation speed in the prior art is generally more than 200 r/min.)
Experimental example 6
Experimental example 6 the working conditions were as in Experimental example 5 except that the digestion time was 6 hours.
Note: the raw materials used in the experimental examples 1 to 6 were 1.5 to 2.5 Kg.
Experiment 1 comparative experiment of fish oil yield in example 4 and experiment examples 1 to 6
The yields of fish oil finally obtained in examples 4 and 1 to 6 were calculated, and the results are shown in table 1, and the calculation formula is as follows:
TABLE 1 comparison of fish oil yields
As can be seen from Table 1, the method can effectively improve the yield of the fish oil, and is obviously superior to the autolytic enzyme method and the protease method. An excess of lipase is disadvantageous and may promote emulsification of the system and prevent the extraction of fats and oils.
It should be noted that the protease method (experimental example 6) extended the reaction time to 6 hours, and the yield was rather decreased, and it is presumed that the precipitation was slow and the emulsification was active as the time was extended, and finally the speed of emulsification exceeded the precipitation speed, and the yield was reversed. If the enzymolysis time is continued to be prolonged, the emulsification will decline with the further reduction of the molecular weight of the protein in the system, and the oil will be separated out again, but the reaction lasts for more than 18 hours, the oil is dark brown in color and luster, bad in smell, and the acid value and the peroxide value can not meet the national requirements.
Experiment 2 acid value and peroxide value of fish oil finally obtained in example 4 and experiment examples 4-6 were compared
The acid value of the fish oil finally obtained in example 4 and experimental examples 4 to 6 was measured by GB/T5530-2005 using the hot ethanol assay, and the peroxide value of the fish oil finally obtained in example 4 and experimental examples 4 to 6 was measured by GB/T5538-2005 using the sodium thiosulfate titration method, and the influence of the patented method and the conventional enzymatic method on the quality (rancidity degree) of the fish oil during extraction of the visceral fish oil was evaluated.
The results are shown in Table 2.
TABLE 2 comparison table of acid value and peroxide value of fish oil
As can be seen from Table 2, under the same reaction time conditions, the acid value and the peroxide value of the method are lower than those obtained by the protease method, and rancidity of the oil and fat in the extraction process can be inhibited to a certain extent. Experimental example 6 the reaction time was 6 hours, and the degree of rancidity was severe.
Experiment 3 comparison of sensory Properties of fish oil obtained in example 4 and experiment examples 4 to 6
And (4) performing quality identification on the viscera crude fish oil through sensory evaluation scores. And randomly selecting 15 professional technicians and 15 non-professional technicians to respectively score the odor and the appearance of the fish oil finally obtained in the embodiment 4 and the experimental examples 4-6, superposing the two components to calculate an average score, and taking two effective numbers to obtain a sensory score. The scoring criteria are shown in Table 3, and reference is made to the scoring criteria
The grading standard SC/T3502-2016 of crude fish oil is shown in Table 4, and the scoring results are shown in Table 5.
TABLE 4 grading Standard Table of crude Fish oils
TABLE 5 sensory evaluation score sheet
As can be seen from Table 5, the odor and the appearance of the product of example 4 of the present invention are superior to those of the experimental examples, and the product has bright light orange color, good transparency and no rancidity odor; the experimental example 4 using the protease alone has a good color, but has a turbid texture and poor transparency, and has rancid odor; experimental example 5 in addition to experimental example 4, the increase in stirring resulted in an increase in the yield of fats and oils, but further deterioration in odor and appearance; experimental example 6 the extraction time is long, a large amount of micromolecular pigment and flavor substances are released from protein tissues and dissolved into grease, the color is dark and turbid, and the rancid smell is prominent.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (6)
1. A preparation method of salmon viscera fish oil is characterized by comprising the steps of raw material pretreatment, first-stage enzymolysis and second-stage enzymolysis;
the first-stage enzymolysis is lipase enzymolysis, and calcium chloride is added in the enzymolysis process;
the second-stage enzymolysis is protease enzymolysis.
2. The preparation method of claim 1, wherein the working conditions of the first stage of enzymolysis are as follows:
mixing the pretreated visceral tissue with water according to a mass ratio of 1: 0.5-1: 2, and adding calcium chloride; adding the lipase in an amount of 100-160 u/g by mass of a reaction system, carrying out enzymolysis at 40-55 ℃ and pH of 7.0-8.5 for 1-2 h, inactivating the enzyme, cooling to below 25 ℃, and carrying out centrifugal separation to obtain the upper-layer crude fish oil.
3. The production method according to claim 2, wherein the calcium chloride is added in an amount of 1.0 to 1.3wt% based on the mass of the added water.
4. The preparation method according to claim 1, wherein the working conditions of the secondary enzymolysis are as follows:
adding protease into a system left after the first-stage enzymolysis crude fish oil is separated, wherein the addition amount of the protease is 1500-2500 u/g based on the mass of the system left, the enzymolysis temperature is 45-55 ℃, the pH value is 7.0-8.0, cooling to below 25 ℃ after enzymolysis is carried out for 1-2 h, and carrying out centrifugal separation to obtain the upper-layer crude fish oil.
5. The method according to claim 4, wherein the protease is an animal protease.
6. The method of claim 1, wherein the raw material pretreatment comprises removal of gall bladder, air bladder and feces from the salmon viscera.
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