CN112625786A - Preparation method of tuna oil - Google Patents
Preparation method of tuna oil Download PDFInfo
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- CN112625786A CN112625786A CN202011623626.XA CN202011623626A CN112625786A CN 112625786 A CN112625786 A CN 112625786A CN 202011623626 A CN202011623626 A CN 202011623626A CN 112625786 A CN112625786 A CN 112625786A
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/02—Pretreatment
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- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/10—Production of fats or fatty oils from raw materials by extracting
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/10—Production of fats or fatty oils from raw materials by extracting
- C11B1/106—Production of fats or fatty oils from raw materials by extracting using ultra-sounds
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- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B3/00—Refining fats or fatty oils
- C11B3/001—Refining fats or fatty oils by a combination of two or more of the means hereafter
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B3/00—Refining fats or fatty oils
- C11B3/005—Refining fats or fatty oils by wave energy or electric current, e.g. electrodialysis
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
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- C11B3/00—Refining fats or fatty oils
- C11B3/02—Refining fats or fatty oils by chemical reaction
- C11B3/06—Refining fats or fatty oils by chemical reaction with bases
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6409—Fatty acids
- C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
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Abstract
The invention relates to the technical field of tuna processing, in particular to a preparation method of tuna oil. The method comprises the steps of raw material pretreatment, enzymolysis and separation, and specifically, the method adopts twice, and adopts the complex enzyme under the vacuum condition in the enzymolysis, so that the prepared fish oil has fewer impurities and bright color. The preparation method can promote the grease in the tissues to be fully released, thereby improving the extraction rate of the fish oil; meanwhile, the method reduces oxidation in the extraction process as much as possible, thereby avoiding quality reduction caused by oxidation; in addition, the good quality is achieved by inhibiting oxidation and actively removing fishy smell.
Description
Technical Field
The invention relates to the technical field of tuna processing, in particular to a preparation method of tuna oil.
Background
The tuna meat is tender and delicious, has high protein content, is rich in protein, fat, vitamin A, D and trace elements, and is also rich in polyunsaturated fatty acids with bioactivity such as docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA); meanwhile, the methionine, the taurine, the mineral substances and the vitamins are rich in content, and the food is a green pollution-free healthy food recommended by the international nutrition association. EPA and DHA mainly have the effects of inhibiting platelet aggregation, reducing thrombosis, lowering cholesterol and preventing cardiovascular and cerebrovascular diseases; anti-inflammatory, anticancer, and immunity enhancing effects; enhancing nervous system function, improving intelligence, strengthening brain, preventing senile dementia, and protecting vision.
Tuna is a very important fish with a very high nutritional value. Fish oil is an important way for people to obtain fish-affecting ingredients.
The prior preparation method of the fish oil has the problems that the purity of the extracted fish oil is not high and the yield cannot be guaranteed, and meanwhile, the fish oil needs to be decolored, deodorized and subjected to impurity removal in the process of extracting the fish oil, so that the fishy smell cannot be completely removed due to incomplete operation, and the quality of the fish oil is affected.
The above problems are caused by the fact that the extraction process of fish oil is not mature enough. Therefore, how to design a method for extracting fish oil from tuna is a key point for obtaining high-quality fish oil.
Disclosure of Invention
The invention aims to provide a preparation method of tuna oil, so that a high-yield and high-quality tuna oil product can be prepared by using easily-realized operation.
The technical scheme adopted by the invention for solving the technical problems is as follows:
the preparation method of the tuna oil comprises the steps of raw material pretreatment, enzymolysis and separation, and specifically comprises the following steps:
firstly, preprocessing, specifically removing viscera of tuna, and then crushing the tissues of the rest part of the tuna into 1cm square fragments; after pretreatment, carrying out cleaning and soaking treatment by using plasma water, and carrying out microwave treatment for 3-5min after draining;
cleaning and soaking with plasma water, and performing microwave treatment to sterilize tuna tissues and preliminarily destroy the molecular structure of fish;
then soaking the mixture for 0.5 to 2 hours by using a sodium hydroxide solution with the concentration of 1 to 3 percent, and then draining; and by utilizing alkalinity, partial substances which are easy to generate rancidity are removed, and the rancidity is prevented from being deepened in the enzymolysis process.
Performing primary enzymolysis, namely performing primary enzymolysis on the tuna tissues by using enzymolysis liquid; adopting immobilized lipase, wherein the addition amount of the lipase is 80-130u/g, and the weight ratio of the enzymolysis liquid to the tissue of the tuna is 1: 3-4; adding 2-3.5% of calcium chloride by mass into the enzymolysis liquid, and keeping the pressure at 5-20kpa under vacuum condition; the enzymolysis temperature is 30-40 ℃; keeping pH at 6.5-7.0, and performing enzymolysis for 5-8 hr to extinguish fire; filtering to remove residue to obtain primary enzymolysis solution;
carrying out secondary enzymolysis on the primary enzymolysis liquid, adding composite protease and partial glyceride lipase into the primary enzymolysis liquid, wherein the addition amount of the composite protease is 800u/g and the addition amount of the partial glyceride lipase is 200u/g, and the enzymolysis temperature is increased to 52-55 ℃; adjusting the pH value to 7.5-8.0; keeping the pressure at 5-20kpa under vacuum condition, and extinguishing fire after enzymolysis for 1-2 h; obtaining secondary enzymolysis liquid;
in the conventional enzymatic method for extracting the fish oil, only a single protease is used for disintegrating protein tissues to release the fish oil, however, the fish oil is very easy to emulsify after being subjected to enzymolysis to a certain degree, so that part of the fish oil is coated in an emulsion, and the yield of the fish oil is reduced in the subsequent separation;
vacuum is adopted in the secondary enzymolysis, so that the proportion of fish oil embedded in the emulsion is reduced, and the yield of the fish oil is improved.
When the method is applied to the extraction of the internal organs of the salmon, the reaction condition is difficult to control, and the serious emulsification is generated after the enzymolysis for a certain time,
the fish oil which is originally precipitated is combined with protein emulsion and protein liquid in a large amount in a short time, so that the ideal yield of the fish oil is difficult to achieve.
If the amount of enzyme used is increased after emulsification, the enzymolysis is continued for a long time to ensure that most of the protein tissue is enzymolyzed into small peptides, and an emulsification system
Is damaged, the emulsified fish oil can be partially separated out, but the whole fish oil is dark and dark brown, which brings great convenience for subsequent processing
It is difficult to use the polyunsaturated fatty acid as a source of polyunsaturated fatty acids for pharmaceutical and high-end foods. If the enzymolysis is stopped before emulsification occurs, the
The fish oil yield is very low.
Heating the secondary enzymolysis liquid to 60-70 ℃, and adjusting the pH to 8.0-9.0; preserving heat for 1-2h, and then cooling to below 30 ℃;
and centrifuging to obtain the upper layer of fish oil.
Preferably, in centrifugation, vacuum centrifugation is used, maintaining a vacuum of less than 1 kpa.
Preferably, the residue obtained by filtering in one enzymolysis step is involved in the next enzymolysis step.
Preferably, the compound protease is trypsin, papain, subtilisin and flavourzyme which are compounded in a ratio of 6:4:3: 1.
The invention has the beneficial effects that: the fish oil prepared by the preparation method disclosed by the application has fewer impurities and bright color. The preparation method can promote the grease in the tissues to be fully released, thereby improving the extraction rate of the fish oil; meanwhile, the method reduces oxidation in the extraction process as much as possible, thereby avoiding quality reduction caused by oxidation; in addition, the good quality is achieved by inhibiting oxidation and actively removing fishy smell.
Detailed Description
Representative embodiments will now be further refined. It should be understood that the following description is not intended to limit the embodiments to one preferred embodiment. On the contrary, it is intended to cover alternatives, modifications, and equivalents as may be included within the spirit and scope of the embodiments as defined by the appended claims.
In the following detailed description, while these embodiments are described in sufficient detail to enable those skilled in the art to practice the embodiments, it is to be understood that these examples are not limiting, such that other examples may be used and that corresponding modifications may be made without departing from the spirit and scope of the embodiments.
Specifically, the application provides a preparation method of tuna oil, which comprises the steps of raw material pretreatment, enzymolysis and separation, and specifically comprises the following detailed steps:
firstly, preprocessing, specifically removing viscera of tuna, and then crushing the tissues of the rest part of the tuna into 1cm square fragments; after pretreatment, plasma water is adopted for cleaning and soaking treatment, and microwave treatment is carried out for 3-5min after draining.
Soaking in 1-3% sodium hydroxide solution for 0.5-2 hr, and draining.
Performing primary enzymolysis, namely performing primary enzymolysis on the tuna tissues by using enzymolysis liquid; adopting immobilized lipase, wherein the addition amount of the lipase is 80-130u/g, and the weight ratio of the enzymolysis liquid to the tissue of the tuna is 1: 3-4; adding 2-3.5% of calcium chloride by mass into the enzymolysis liquid, and keeping the pressure at 5-20kpa under vacuum condition; the enzymolysis temperature is 30-40 ℃; keeping pH at 6.5-7.0, and performing enzymolysis for 5-8 hr to extinguish fire; filtering to remove residue to obtain primary enzymolysis solution; the dregs obtained by filtering in one enzymolysis participate in the next enzymolysis.
Carrying out secondary enzymolysis on the primary enzymolysis liquid, adding composite protease and partial glyceride lipase into the primary enzymolysis liquid, wherein the addition amount of the composite protease is 800u/g and the addition amount of the partial glyceride lipase is 200u/g, and the enzymolysis temperature is increased to 52-55 ℃; adjusting the pH value to 7.5-8.0; keeping the pressure at 5-20kpa under vacuum condition, and extinguishing fire after enzymolysis for 1-2 h; obtaining secondary enzymolysis liquid; the compound protease is composed of trypsin, papain, subtilisin and flavourzyme in a ratio of 6:4:3: 1.
Heating the secondary enzymolysis liquid to 60-70 ℃, and adjusting the pH to 8.0-9.0; preserving heat for 1-2h, and then cooling to below 30 ℃;
and (4) performing centrifugal separation, namely performing vacuum centrifugal separation, and keeping the vacuum of less than 1kpa to obtain the upper layer, namely the fish oil.
Example 1: a preparation method of tuna oil comprises the following detailed steps:
firstly, preprocessing, specifically removing viscera of tuna, and then crushing the tissues of the rest part of the tuna into 1cm square fragments; after pretreatment, plasma water is adopted for cleaning and soaking treatment, and microwave treatment is carried out for 3min after draining.
Then soaking the mixture for 2 hours in a sodium hydroxide solution with the concentration of 1 percent, and then draining the mixture.
Then, carrying out primary enzymolysis on the tuna tissues by adopting enzymolysis liquid; the enzymolysis liquid adopts immobilized lipase, the addition amount of the lipase is 130u/g, and the enzymolysis liquid is prepared according to the weight ratio of the enzymolysis liquid to the tissue of the tuna being 1: 3; adding calcium chloride with the mass fraction of 3.5% into the enzymolysis liquid, and keeping the pressure at 10kpa under vacuum condition; the enzymolysis temperature is 40 ℃; keeping pH at 6.5-7.0 weak acidity in the whole process, and extinguishing fire after enzymolysis for 8 h; filtering to remove residue to obtain primary enzymolysis solution; the dregs obtained by filtering in one enzymolysis participate in the next enzymolysis.
Adding composite protease and partial glyceride lipase into the primary enzymolysis liquid for secondary enzymolysis, wherein the addition amount of the composite protease is 500u/g, the addition amount of the partial glyceride lipase is 200u/g, and the enzymolysis temperature is increased to 52 ℃; adjusting the pH value to be in alkalescence of 7.5-8.0; keeping the pressure at 5kpa under vacuum condition, and extinguishing fire after 2 hours of enzymolysis; obtaining secondary enzymolysis liquid; the compound protease is composed of trypsin, papain, subtilisin and flavourzyme in a ratio of 6:4:3: 1.
Heating the secondary enzymolysis liquid to 70 ℃, adjusting the pH to 8.0-9.0, preserving the heat for 1h, and then cooling to below 30 ℃;
and (4) performing centrifugal separation, namely performing vacuum centrifugal separation, and keeping the vacuum of less than 1kpa to obtain the upper layer, namely the fish oil.
Example 2: a preparation method of tuna oil comprises the following detailed steps:
firstly, preprocessing, specifically removing viscera of tuna, and then crushing the tissues of the rest part of the tuna into 1cm square fragments; after pretreatment, plasma water is adopted for cleaning and soaking treatment, and microwave treatment is carried out for 5min after draining.
Then soaking in 3% sodium hydroxide solution for 0.5h, and draining.
Then, carrying out primary enzymolysis on the tuna tissues by adopting enzymolysis liquid; the enzymolysis liquid adopts immobilized lipase, the addition amount of the lipase is 80u/g, and the enzymolysis liquid is prepared according to the weight ratio of the enzymolysis liquid to the tissue of the tuna being 1: 3; adding calcium chloride with the mass fraction of 2% into the enzymolysis liquid, and keeping the pressure at 5kpa under vacuum condition; the enzymolysis temperature is 40 ℃; keeping pH at 6.5-7.0 weak acidity in the whole process, and extinguishing fire after 5h of enzymolysis; filtering to remove residue to obtain primary enzymolysis solution; the dregs obtained by filtering in one enzymolysis participate in the next enzymolysis.
Adding composite protease and partial glyceride lipase into the primary enzymolysis liquid for secondary enzymolysis, wherein the addition amount of the composite protease is 800u/g, the addition amount of the partial glyceride lipase is 200u/g, and the enzymolysis temperature is increased to 52 ℃; adjusting the pH value to be in alkalescence of 7.5-8.0; keeping the pressure at 10kpa under vacuum condition, and extinguishing fire after enzymolysis for 1-2 h; obtaining secondary enzymolysis liquid; the compound protease is composed of trypsin, papain, subtilisin and flavourzyme in a ratio of 6:4:3: 1.
Heating the secondary enzymolysis liquid to 60 ℃, adjusting the pH to 8.0-9.0, preserving the heat for 1-2h, and then cooling to below 30 ℃;
and (4) performing centrifugal separation, namely performing vacuum centrifugal separation, and keeping the vacuum of less than 1kpa to obtain the upper layer, namely the fish oil.
Example 3: a preparation method of tuna oil comprises the following detailed steps:
firstly, preprocessing, specifically removing viscera of tuna, and then crushing the tissues of the rest part of the tuna into 1cm square fragments; after pretreatment, plasma water is adopted for cleaning and soaking treatment, and microwave treatment is carried out for 5min after draining.
Then soaking the mixture for 2 hours in a sodium hydroxide solution with the concentration of 2 percent, and then draining the mixture.
Then, carrying out primary enzymolysis on the tuna tissues by adopting enzymolysis liquid; the enzymolysis liquid adopts immobilized lipase, the addition amount of the lipase is 100u/g, and the enzymolysis liquid is prepared according to the weight ratio of the enzymolysis liquid to the tissue of the tuna being 1: 3; adding calcium chloride with the mass fraction of 3.5% into the enzymolysis liquid, and keeping the pressure at 10kpa under vacuum condition; the enzymolysis temperature is 30 ℃; keeping pH at 6.5-7.0 weak acidity in the whole process, and extinguishing fire after 6h enzymolysis; filtering to remove residue to obtain primary enzymolysis solution; the dregs obtained by filtering in one enzymolysis participate in the next enzymolysis.
Adding composite protease and partial glyceride lipase into the primary enzymolysis liquid for secondary enzymolysis, wherein the addition amount of the composite protease is 700u/g, the addition amount of the partial glyceride lipase is 150u/g, and the enzymolysis temperature is increased to 53 ℃; adjusting the pH value to be in alkalescence of 7.5-8.0; extinguishing fire after enzymolysis for 1.5 h; and keeping the pressure at 15kpa under vacuum condition to obtain secondary enzymolysis liquid; the compound protease is composed of trypsin, papain, subtilisin and flavourzyme in a ratio of 6:4:3: 1.
Heating the secondary enzymolysis liquid to 67 ℃, adjusting the pH to 8.0-9.0, preserving the heat for 2 hours, and then cooling to below 30 ℃;
and (4) performing centrifugal separation, namely performing vacuum centrifugal separation, and keeping the vacuum of less than 1kpa to obtain the upper layer, namely the fish oil.
The results of extraction, purity, fishy smell and color are shown in the table below for the above examples.
| Extraction rate | Purity of | Smell(s) | Appearance of the product | |
| Example 1 | 92% | 95.3% | 8.5 | 9.3 |
| Example 2 | 90% | 96.8% | 9.0 | 9.2 |
| Example 3 | 91% | 94.9% | 8.7 | 9.6 |
Odor scoring criteria: 1-3 has strong fishy smell and rancid taste; 4-7 has slight fishy smell and is not obvious in rancidity taste; no fishy smell, no rancidity smell and fragrant smell for 8-10 households.
Appearance: 1-3 earthy yellow and turbid with obvious precipitation; 4-7 orange, slightly turbid, slightly precipitated; 8-10 transparent egg yolk without precipitate.
As can be seen from the above table, the extracted fish oil of the present invention has a better appearance, is transparent and light yellow, and has no fishy smell.
The prepared fish oil was subjected to a performance test according to SC/T3502, and the test structure is shown in the following table:
| item | Example 1 | Example 2 | Example 3 |
| Water and volatile matter% | ≤0.2 | ≤0.2 | ≤0.2 |
| Acid number (calculated as KOH), mg/g | ≤3.0 | ≤3.0 | ≤3.0 |
| Peroxide number, meq/kg | ≤10.0 | ≤10.0 | ≤10.0 |
| Anisidine number | ≤20.0 | ≤20.0 | ≤20.0 |
| Iodine value, g/100g | ≥140 | ≥140 | ≥140 |
| Insoluble impurities,% of | ≤0.1 | ≤0.1 | ≤0.1 |
| Unsaponifiable matter,% | ≤3.0 | ≤3.0 | ≤3.0 |
As can be seen from the above table, the quality of the fish oil prepared by the method is far higher than the quality requirement of crude oil, and the basic requirement of refined fish oil can be met.
For purposes of explanation, specific nomenclature is used in the above description to provide a thorough understanding of the described embodiments. It will be apparent, however, to one skilled in the art that these specific details are not required in order to practice the embodiments described above. Thus, the foregoing descriptions of specific embodiments described herein are presented for purposes of illustration and description. It is not intended to be exhaustive or to limit the embodiments to the precise form disclosed. It will be apparent to those skilled in the art that certain modifications, combinations, and variations can be made in light of the above teachings.
Claims (6)
1. The preparation method of the tuna oil comprises the steps of raw material pretreatment, enzymolysis and separation, and is characterized in that: specifically, the method comprises the following steps:
firstly, preprocessing, specifically removing viscera of tuna, and then crushing the tissues of the rest part of the tuna into 1cm square fragments; after pretreatment, carrying out cleaning and soaking treatment by using plasma water, and carrying out microwave treatment for 3-5min after draining;
then soaking the mixture for 0.5 to 2 hours by using a sodium hydroxide solution with the concentration of 1 to 3 percent, and then draining;
performing primary enzymolysis, namely performing primary enzymolysis on the tuna tissues by using enzymolysis liquid; adopting immobilized lipase, wherein the addition amount of the lipase is 80-130u/g, and the weight ratio of the enzymolysis liquid to the tissue of the tuna is 1: 3-4; adding 2-3.5% of calcium chloride by mass into the enzymolysis liquid, and keeping the pressure at 5-20kpa under vacuum condition; the enzymolysis temperature is 30-40 ℃; keeping pH at 6.5-7.0, and performing enzymolysis for 5-8 hr to extinguish fire; filtering to remove residue to obtain primary enzymolysis solution;
carrying out secondary enzymolysis on the primary enzymolysis liquid, adding composite protease and partial glyceride lipase into the primary enzymolysis liquid, wherein the addition amount of the composite protease is 800u/g and the addition amount of the partial glyceride lipase is 200u/g, and the enzymolysis temperature is increased to 52-55 ℃; adjusting the pH value to 7.5-8.0; keeping the pressure at 5-20kpa under vacuum condition, and extinguishing fire after enzymolysis for 1-2 h; obtaining secondary enzymolysis liquid;
heating the secondary enzymolysis liquid to 60-70 ℃, and adjusting the pH to 8.0-9.0; preserving heat for 1-2h, and then cooling to below 30 ℃;
and centrifuging to obtain the upper layer of fish oil.
2. The method for preparing tuna oil according to claim 1, wherein: in centrifugation, vacuum centrifugation is used, maintaining a vacuum of less than 1 kpa.
3. The method for preparing tuna oil according to claim 1, wherein: the dregs obtained by filtering in one enzymolysis participate in the next enzymolysis.
4. The method for preparing tuna oil according to claim 1, wherein: the compound protease is compounded by trypsin, papain, subtilisin and flavourzyme in a ratio of 6:4:3: 1.
5. The method for preparing tuna oil according to claim 1, wherein: in the first enzymolysis, the vacuum condition with the pressure of 5-10kpa is maintained.
6. The method for preparing tuna oil according to claim 1, wherein: in the secondary enzymolysis, the vacuum condition with the pressure of 5-8kpa is kept.
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