CN107629865A - A kind of enzymolysis solvent extraction process of squid liver grease - Google Patents
A kind of enzymolysis solvent extraction process of squid liver grease Download PDFInfo
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- CN107629865A CN107629865A CN201711092325.7A CN201711092325A CN107629865A CN 107629865 A CN107629865 A CN 107629865A CN 201711092325 A CN201711092325 A CN 201711092325A CN 107629865 A CN107629865 A CN 107629865A
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- enzymolysis
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Abstract
The invention discloses a kind of enzymolysis solvent extraction process of squid liver grease, and it passes through step：（I）Squid viscera is thawed and cuts into particle and adds water；（II）Alkali protease and papain, regulation pH enzymolysis, heat inactivation are added to the squid viscera mixture；（III）Digest mixture and centrifuge the first oil product and lower floor's enzymolysis liquid；（IV）N-hexane and stirring will be added in lower floor's enzymolysis liquid, oil phase is collected and evaporates recovery and obtain the second oil product；（V）The second oil product that step IV is obtained is vacuumized and stirred；（VI）The first oil product that the oil product and step III that blend step V is obtained obtain, obtains final products.The method of the present invention realizes higher oil recovery rate, and obtain safer squid liver grease without using the solvent of toxic side effect by using enzymolysis and two step separation methods.
Description
Technical field
The present invention relates to the refined of fatty oil, especially a kind of enzymolysis solvent extraction process of squid liver grease.
Background technology
It is known that in squid process, while squid body weight 15-20% squid viscera can be produced.This part pair
Product mainly has two kinds of processing modes at present：Crush after drying and sold as fish meal, or for extracting squid liver grease.
And containing abundant grease in squid liver, unsaturated fatty acid content is very high wherein in grease, and EPA and DHA content account for fat
35% or so of sour total amount, therefore be the raw material of good extraction squid grease.
At present, the squid in the methods of typically taking squeezing, boiling, enzymolysis, supercritical fluid, solvent extraction squid viscera
Liver grease.In these methods, the rate of recovery of squeezing and boiling method is very low；It is left that the rate of recovery of enzyme solution is likely to be breached 70%
The right side, and the quality of its extraction grease is higher；The rate of recovery of extract by solvents can reach more than 90%, but extract what is obtained
The quality of grease is relatively low, and easily because residual solvent causes the toxic side effect of grease；Supercritical fluid processes are to equipment
It is required that it is higher, it is unfavorable for small-scale production.
The content of the invention
In order to overcome deficiency of the prior art, it is an object of the invention to provide a kind of rate of recovery it is higher, independent of having
The enzymolysis separation method of the gentle squid liver grease of solvent, process conditions.
The technical solution adopted for the present invention to solve the technical problems is：A kind of enzymolysis solvent extraction of squid liver grease
Method, it is characterised in that：Described method passes through following steps：
（I）By squid viscera defrosting and 10-20mm particle is cut into, the water of the weight such as addition；
（II）The squid viscera mixture obtained to step I adds alkali protease and papain to 1500-2000U/mL
And 2000-2600U/mL, using sodium carbonate or salt acid for adjusting pH to 9.0-9.5, and 2-4 is digested under 40-45 degrees Celsius
Hour, heat inactivation；
（III）The enzymolysis mixture that step II is obtained is centrifuged into 15-30 minutes in 1500-2000 revs/min of centrifuge, and
Separate the first oil product and lower floor's enzymolysis liquid；
（IV）Weight 12-20% n-hexane will be added in the lower floor's enzymolysis liquid obtained in step III, and stir 10-20 points
Clock, collect oil phase and evaporate n-hexane and reclaim to obtain the second oil product；
（V）The second oil product that step IV is obtained vacuumizes under 0-15 degrees Celsius reaches 0.05-0.1 atmospheric pressure, and
Stir 15-30 minutes；
（VI）The first oil product that the oil product and step III that blend step V is obtained obtain, obtains final products.
Mixed between currently preferred method, described step I and II, in addition to by the obtained squid visceras of step I
Compound between 40-45 degrees Celsius stir 1-2 hours the step of.
In currently preferred aspect, described step II, the squid viscera mixture obtained to step I adds alkalescence
Protease and papain are to 1700U/mL and 2200U/mL, using sodium carbonate or salt acid for adjusting pH to 9.2, and 42
Digested 3.2 hours under degree Celsius.
In currently preferred aspect, described step III, 4.5-5.5 is arrived into pH regulations before centrifugation.
In currently preferred aspect, described step III, by the enzymolysis mixture that step II is obtained 1800 turns/
Centrifuged 22 minutes in the centrifuge divided, and separate the first oil product and lower floor's enzymolysis liquid.
In currently preferred aspect, described step IV, weight will be added in the lower floor's enzymolysis liquid obtained in step III
The n-hexane of amount 15%, and stir 18 minutes, collect oil phase and evaporate n-hexane and reclaim to obtain the second oil product.
In currently preferred aspect, the second oil product that step IV is obtained vacuumizes at 4 deg. celsius to be reached
0.08 atmospheric pressure, and stir 22 minutes.
In currently preferred aspect, described step VI, in the obtained oil products of blend step V and step III
It is described after the first obtained oil product, in addition to the step of the oil product of mixing is washed using cleaning solution
Cleaning solution includes the sodium citrate that mass fraction is 0.5-1.5% and 0.3-0.5% EDETATE DISODIUM.
In currently preferred aspect, described step VI, in the obtained oil products of blend step V and step III
It is described after the first obtained oil product, in addition to the step of the oil product of mixing is washed using cleaning solution
Cleaning solution includes the sodium citrate and 0.34% EDETATE DISODIUM that mass fraction is 0.90%.
The method of the present invention realizes higher oil recovery rate by using enzymolysis and two step separation methods, and by
Safer squid liver grease has been obtained in the solvent without using toxic side effect.The method process conditions temperature of the present invention
It is feasible with, advanced technology, operation, it can be widely applied in the preparation of fish oil class health products.
Embodiment
Unless additionally illustrating, the papain and alkali protease in following embodiment are purchased from the permanent Hua Daosheng in east
Thing Science and Technology Ltd..Two kinds of enzymes are 800,000 U/ grams of dry powder.When adding, add water that 100,000 U/mL are made in the dry powder first
Enzyme solutions, then the enzyme solutions of certain volume are added in seafood mixture, reach the amount of expectation.
Unless additionally illustrating, the squid viscera of double centner is used in following embodiment and comparative example.All realities
The step I for applying example and comparative example is general step, and is carried out in a big food processing container, so all implementation
The step I of example and comparative example result is identical, takes identical to originate in subsequent steps.
Embodiment 1
In the present embodiment, a kind of enzymolysis solvent extraction process of squid liver grease, it passes through following steps：
（I）By squid viscera defrosting and 10-20mm particle is cut into, the water of the weight such as addition；
（II）The squid viscera mixture obtained to step I add alkali protease and papain to 1700U/mL and
2200U/mL, using sodium carbonate or salt acid for adjusting pH to 9.2, and digested 3.2 hours under 42 degrees Celsius, heat inactivation；
（III）The enzymolysis mixture that step II is obtained is adjusted pH to 5.0 before centrifugation, in 1800 revs/min of centrifuge
Middle centrifugation 22 minutes, and separate the first oil product and lower floor's enzymolysis liquid；
（IV）The n-hexane of weight 15% will be added in the lower floor's enzymolysis liquid obtained in step III, and stirred 18 minutes, collected
Oil phase and evaporate n-hexane and reclaim to obtain the second oil product；
（V）The second oil product that step IV is obtained vacuumizes at 4 deg. celsius reaches 0.08 atmospheric pressure, and stirs 22 points
Clock；
（VI）The first oil product that the oil product and step III that blend step V is obtained obtain, obtains final products.
Embodiment 2
In the present embodiment, a kind of enzymolysis solvent extraction process of squid liver grease, it passes through following steps：
（I）By squid viscera defrosting and 10-20mm particle is cut into, the water of the weight such as addition；
（II）The squid viscera mixture obtained to step I add alkali protease and papain to 1500U/mL and
2000U/mL, using sodium carbonate or salt acid for adjusting pH to 9.5, and digested 4 hours under 45 degrees Celsius, heat inactivation；
（III）The enzymolysis mixture that step II is obtained is adjusted pH to 4.5 before centrifugation, in 2000 revs/min of centrifuge
Middle centrifugation 15 minutes, and separate the first oil product and lower floor's enzymolysis liquid；
（IV）The n-hexane of weight 20% will be added in the lower floor's enzymolysis liquid obtained in step III, and stirred 10 minutes, collected
Oil phase and evaporate n-hexane and reclaim to obtain the second oil product；
（V）The second oil product that step IV is obtained vacuumizes under 15 degrees Celsius reaches 0.05 atmospheric pressure, and stirs 15
Minute；
（VI）The first oil product that the oil product and step III that blend step V is obtained obtain, obtains final products.
Embodiment 3
In the present embodiment, a kind of enzymolysis solvent extraction process of squid liver grease, it passes through following steps：
（I）By squid viscera defrosting and 10-20mm particle is cut into, the water of the weight such as addition；
（II）The squid viscera mixture obtained to step I add alkali protease and papain to 2000U/mL and
2600U/mL, using sodium carbonate or salt acid for adjusting pH to 9.0, and digested 2 hours under 40 degrees Celsius, heat inactivation；
（III）The enzymolysis mixture that step II is obtained is adjusted pH to 5.5 before centrifugation, in 1500 revs/min of centrifuge
Middle centrifugation 30 minutes, and separate the first oil product and lower floor's enzymolysis liquid；
（IV）The n-hexane of weight 12% will be added in the lower floor's enzymolysis liquid obtained in step III, and stirred 20 minutes, collected
Oil phase and evaporate n-hexane and reclaim to obtain the second oil product；
（V）The second oil product that step IV is obtained vacuumizes under 0 degree Celsius reaches 0.1 atmospheric pressure, and stirs 30 points
Clock；
（VI）The first oil product that the oil product and step III that blend step V is obtained obtain, obtains final products.
Embodiment 4
The method and embodiment 1 used in the present embodiment is essentially identical, and its difference is：Between described step I and II, also wrap
Include the squid viscera mixture that step I is obtained stirred 1.6 hours between 42 degrees Celsius the step of.
Embodiment 5
The method and embodiment 1 used in the present embodiment is essentially identical, and its difference is：In described step VI, in mixing step
After the first oil product that the oil product and step III that rapid V is obtained obtain, in addition to the oil product use of mixing washed
The step of liquid is washed is washed, described cleaning solution includes the sodium citrate and 0.34% EDETATE SODIUM that mass fraction is 0.90%
Salt.
Embodiment 6
The method and embodiment 1 used in the present embodiment is essentially identical, and its difference is：Between described step I and II, also wrap
Include the squid viscera mixture that step I is obtained stirred 2 hours between 40 degrees Celsius the step of；In described step VI,
After the first oil product that the oil product and step III that blend step V is obtained obtain, in addition to the oil product by mixing
The step of being washed using cleaning solution, described cleaning solution include the sodium citrate and 0.5% that mass fraction is 0.50%
EDETATE DISODIUM.
Comparative example 1
In this comparative example, the method for extraction squid liver grease includes：
（I）By squid viscera defrosting and 10-20mm particle is cut into, the water of the weight such as addition；
（II）N-hexane-isopropanol (3: 2) is solvent, the mL/g of liquid ratio 10, the min of extraction time 30, temperature 60 C, and is carried
Ultrasonication is applied to squid viscera mixture during taking.
After extraction finishes, the isopropanol content by gas chromatography analysis in squid liver grease, find wherein
The isopropanol content contained is 0.45-0.52 wt.%.
Comparative example 2
The method and embodiment 1 used in this comparative example is essentially identical, and its difference is：Step VI-V is not performed.
Comparative example 3
The method and embodiment 1 used in this comparative example is essentially identical, and its difference is：Step V is not performed.
The rate of recovery is carried out to the squid liver grease extracted in embodiment 1-6 and comparative example 1-3 below and peroxidating tries
Test.
First, squid liver oil recovery rate is tested
In this experiment, embodiment 1-6 and comparative example 1-3 squid liver grease are weighed, are as a result shown in table 1.
Table 1：Embodiment 1-:6 and comparative example 1-3 squid liver grease weight.
| Test group | Squid liver grease weight/kg |
| Embodiment 1 | 18.19 |
| Embodiment 2 | 18.32 |
| Embodiment 3 | 18.63 |
| Embodiment 4 | 20.23 |
| Embodiment 5 | 18.01 |
| Embodiment 6 | 17.85 |
| Comparative example 1 | 19.83 |
| Comparative example 2 | 15.21 |
| Comparative example 3 | 18.54 |
It can see from table 1, when being extracted using organic solvent（Comparative example 1）, obtained very high-recovery
Squid liver grease.When only using enzymolysis product to carry out single stage centrifugation, very low-producing squid liver oil has been obtained
Fat（Comparative example 2）.When aiding in n-hexane recovery, grease yield has been lifted（Comparative example 3）.In embodiment 1-3 and embodiment
In 5-6, the grease yield of very high content has also been obtained.Applicant have surprisingly discovered that internal organ mixture is allowed to exist before enzymolysis
Voluntarily degraded under initial condition after a period of time, extraction efficiency can be lifted.
2nd, squid liver oil peroxidation value is tested
After the extraction grease of each group stores 1 month, peroxide value measurement experiment is carried out, measuring method is according to GB/T5338-
Method described by 2005 is carried out.Specific method step includes：2.00 grams of oil samples are weighed, are placed in 250mL iodine flasks,
And 50 milliliters of isooctane-glacial acetic acid solutions are added, the shake that closes the lid is until sample dissolves.Add 0.5mL saturation KIs
Solution, closeing the lid makes its reaction, and the time is 1 minute, is at least shaken the bottle during this period 3 times.30 milliliters are added immediately after
Distilled water, and titrated using 0.01M hypo solution, until yellow almost disappears.Add about 0.5mL starch water
Solution, continue to titrate, constantly shaken in titration process, until blueness disappears.Measurement result is scaled mM every kilogram of expression.
Each group of peroxide value measurement result is shown in table 2.
Table 2：Embodiment 1-6 and comparative example 1-3 peroxide value measurement result.
It can be seen that, the liver grease for extracting to obtain in comparative example 1 has very high peroxide value measurement result from table 2, this
Show that the liver grease experienced complex and fierce chemical treatment in processing procedure, cause caused oil quality not
Height, its freshness can not meet the requirements.The grease that peroxide value is told somebody what one's real intentions are has been obtained in embodiment 1-3.By further
After washing, the peroxide value of grease create a further reduction.This shows the processing by embodiment 5-6, eliminates part meeting
Cause the oxidized material of grease so that the quality of grease increases.
Claims (9)
- A kind of 1. enzymolysis solvent extraction process of squid liver grease, it is characterised in that：Described method passes through following steps：（I）By squid viscera defrosting and 10-20mm particle is cut into, the water of the weight such as addition；（II）The squid viscera mixture obtained to step I adds alkali protease and papain to 1500-2000U/mL And 2000-2600U/mL, using sodium carbonate or salt acid for adjusting pH to 9.0-9.5, and 2-4 is digested under 40-45 degrees Celsius Hour, heat inactivation；（III）The enzymolysis mixture that step II is obtained is centrifuged into 15-30 minutes in 1500-2000 revs/min of centrifuge, and Separate the first oil product and lower floor's enzymolysis liquid；（IV）Weight 12-20% n-hexane will be added in the lower floor's enzymolysis liquid obtained in step III, and stir 10-20 points Clock, collect oil phase and evaporate n-hexane and reclaim to obtain the second oil product；（V）The second oil product that step IV is obtained vacuumizes under 0-15 degrees Celsius reaches 0.05-0.1 atmospheric pressure, and Stir 15-30 minutes；（VI）The first oil product that the oil product and step III that blend step V is obtained obtain, obtains final products.
- A kind of 2. enzymolysis solvent extraction process of squid liver grease according to claim 1, it is characterised in that：Described Between step I and II, in addition to the squid viscera mixture that step I is obtained stirs 1-2 hours between 40-45 degrees Celsius Step.
- A kind of 3. enzymolysis solvent extraction process of squid liver grease according to claim 1, it is characterised in that：Described In step II, the squid viscera mixture that obtains to step I add alkali protease and papain to 1700U/mL and 2200U/mL, using sodium carbonate or salt acid for adjusting pH to 9.2, and digested 3.2 hours under 42 degrees Celsius.
- A kind of 4. enzymolysis solvent extraction process of squid liver grease according to claim 1, it is characterised in that：Described In step III, 4.5-5.5 is arrived into pH regulations before centrifugation.
- A kind of 5. enzymolysis solvent extraction process of squid liver grease according to claim 1, it is characterised in that：Described In step III, the enzymolysis mixture that step II is obtained is centrifuged 22 minutes in 1800 revs/min of centrifuge, and separates the One oil product and lower floor's enzymolysis liquid.
- A kind of 6. enzymolysis solvent extraction process of squid liver grease according to claim 1, it is characterised in that：Described In step IV, the n-hexane of weight 15% will be added in the lower floor's enzymolysis liquid obtained in step III, and stirred 18 minutes, collected Oil phase and evaporate n-hexane and reclaim to obtain the second oil product.
- A kind of 7. enzymolysis solvent extraction process of squid liver grease according to claim 1, it is characterised in that：By step The second oil product that IV is obtained vacuumizes at 4 deg. celsius reaches 0.08 atmospheric pressure, and stirs 22 minutes.
- A kind of 8. enzymolysis solvent extraction process of squid liver grease according to claim 1, it is characterised in that：Described In step VI, after the first oil product that the obtained oil products of blend step V and step III are obtained, in addition to will be mixed The step of oil product of conjunction is washed using cleaning solution, described cleaning solution include the lemon that mass fraction is 0.5-1.5% The EDETATE DISODIUM of sour sodium and 0.3-0.5%.
- A kind of 9. enzymolysis solvent extraction process of squid liver grease according to claim 1, it is characterised in that：Described In step VI, after the first oil product that the obtained oil products of blend step V and step III are obtained, in addition to will be mixed The step of oil product of conjunction is washed using cleaning solution, described cleaning solution include the citric acid that mass fraction is 0.90% Sodium and 0.34% EDETATE DISODIUM.
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Cited By (1)
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