CN107629873B - Method for enriching fish oil EPA and DHA through low-temperature crystallization - Google Patents
Method for enriching fish oil EPA and DHA through low-temperature crystallization Download PDFInfo
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Abstract
The invention relates to a method for enriching fish oil EPA and DHA by low-temperature crystallization, belonging to the technical field of aquatic product processing and health products. The method comprises the following steps: pretreating fish oil, adding an organic solvent and a crystallization assistant agent into the fish oil, wherein the mass ratio of the fish oil to the organic solvent to the crystallization assistant agent is 100 (200-1000) to 0.5-2, uniformly mixing, performing low-temperature crystallization, performing vacuum filtration to separate crystals and non-crystallized liquid, and removing the organic solvent and the crystallization assistant agent in the non-crystallized liquid to obtain the enriched fish oil EPA and DHA. The method for enriching the EPA and the DHA in the fish oil by low-temperature crystallization has the advantages of simple equipment, simple and convenient operation, large single treatment capacity, low cost, difficult oxidation, isomerization, aggregation and other reactions of effective components, and better maintenance of the natural activity of the EPA and the DHA.
Description
Technical Field
The invention belongs to the technical field of aquatic product processing and health products, and relates to a method for enriching fish oil EPA and DHA through low-temperature crystallization.
Background
Today, china has grown into the third world EPA and DHA oil market, and DHA + EPA has received new resource food certifications and will be incorporated into DRI, which will help further improve consumer cognition. With the increasing income, education degree and health consciousness of consumers in China year by year, the physiological functions of EPA and DHA, such as preventing atherosclerosis and cardiovascular diseases, preventing and resisting cancer, improving brain function, improving memory, promoting intelligence development of infants and the like, are generally known, and especially the increase of the aged population in China in recent years inevitably brings new market opportunity to EPA and DHA products.
EPA and DHA belong to n-3 series polyunsaturated fatty acids and have important health care effect on human bodies, and fish oil is one of the hot spots of domestic and foreign research for many years because of being rich in n-3 series polyunsaturated fatty acids, especially EPA and DHA.
At present, methods for separating and enriching polyunsaturated fatty acids in fish oil mainly comprise a low-temperature solvent crystallization method, urea inclusion and supercritical CO2Extraction, molecular distillation, enzyme concentration, etc. It is composed ofIn the low-temperature crystallization method, the operation is simple and convenient, the requirements on instruments and equipment are low, the natural activity of the unsaturated fatty acid can be better maintained in a low-temperature environment, the separation effect is better, and the like. Yuanyan et al mixed fatty acids were mixed at a ratio of 1: 3 (acid: solvent) is dissolved in petroleum ether, the solution is cooled to 5-7 ℃, saturated fatty acid is crystallized, unsaturated fatty acid is obtained through suction filtration and distillation, the yield can reach 80%, and a low-temperature crystallization method has a certain enrichment effect (Yuanyan, et al.) research on a method for separating unsaturated fatty acid from flax oil [ J]Proceedings of Liaoning science and technology institute, 2005, 7 (4): 12-13).
So far, many reports about the enrichment of polyunsaturated fatty acids in fish oil exist, and most of the industrialized production of the ethyl ester fish oil adopts a method combining urea inclusion and molecular distillation, and the method has not been broken through for decades. The urea inclusion method has the advantages of simple equipment, mild conditions, low cost, capability of completely retaining the physiological activity of the active ingredients and the like, but needs to consume a large amount of solvent, has the problems of solvent recovery and environmental pollution, and can not separate EPA and DHA. To increase the yield, multiple urea encapsulation processes or a combination of molecular distillation techniques are generally used. The separation and purification process of the molecular distillation technology is relatively complex, for example, multi-stage purification is needed, how to realize continuous feeding, and the influence on material degradation when solid feeding is heated at high temperature is realized. The supercritical extraction technology has the characteristics of low temperature, high pressure and air isolation, is particularly suitable for separating heat-sensitive substances and easily-oxidized substances, can fully protect the quality of fish oil from being damaged, has better product quality, but has high requirements on equipment and high energy consumption,
and the specific process research on the enrichment of fish oil EPA/DHA by the low-temperature crystallization method is not reported, and related condition parameters are more difficult to find as references, so that more enrichment methods are explored, more possibilities are sought, and the method is very important for upgrading and reform of the future fish oil industry.
Disclosure of Invention
The invention aims to solve the problems in the prior art, and provides a method for enriching EPA and DHA in fish oil by low-temperature crystallization, which has the advantages of simple equipment, simple and convenient operation, large single treatment capacity, low cost, difficult oxidation, isomerization, aggregation and other reactions of effective components, and better maintenance of the natural activity of EPA and DHA.
The purpose of the invention can be realized by the following technical scheme: a method for enriching fish oil EPA and DHA through low-temperature crystallization comprises the following steps:
pretreating fish oil, adding an organic solvent and a crystallization assistant agent into the fish oil, wherein the mass ratio of the fish oil to the organic solvent to the crystallization assistant agent is 100 (200-1000) to 0.5-2, uniformly mixing, performing low-temperature crystallization, performing vacuum filtration to separate crystals and non-crystallized liquid, and removing the organic solvent and the crystallization assistant agent in the non-crystallized liquid to obtain the enriched fish oil EPA and DHA.
The method enriches EPA and DHA in the fish oil by a low-temperature crystallization method, is simple and convenient to operate, has low requirements on instruments and equipment, has large single treatment capacity and good separation effect, and can better maintain the natural activity of EPA and DHA in a low-temperature environment. In the low-temperature crystallization process, the addition of the organic solvent improves the liquid phase proportion and the degree of freedom of crystals, so that the later filtration and separation of the crystals from the uncrystallized liquid are easy to carry out. The addition of the crystallization assistant promotes the crystallization, reduces the supercooling degree required by crystallization, improves the crystallization temperature and reduces the crystallization time. The dosage of the organic solvent is related to the content of saturated fatty acid in the raw oil, when the dosage is too small, the whole crystals cannot be separated out, and when the dosage is too large, the raw oil is diluted too much, the granularity and the distribution of crystals are influenced, the granularity of the formed crystals is too small, the too small crystals can absorb more EPA and DHA, the separation efficiency is reduced, and simultaneously the waste of the solvent is caused, so the ratio of the fish oil to the organic solvent is limited within the range. The strength of the crystal structure of the saturated fatty acid formed by freezing is limited, and the crystals and the non-crystallized liquid are separated by vacuum filtration, so that the shearing of the floccule flow can be avoided as much as possible, and the damage to the crystals is reduced.
Preferably, the fish oil is ethyl ester type fish oil.
Preferably, the content of EPA and DHA in the ethyl ester fish oil is 50-70% by mass.
Preferably, the pretreatment is to heat the fish oil at 30-50 ℃ for 5-30 min.
During processing, storage, transportation and the like of fish oil, uneven crystal nuclei are often formed, which is not favorable for the formation of crystal nuclei during low-temperature crystallization, and therefore, the uneven crystal nuclei must be destroyed before low-temperature crystallization. The fish oil can be completely melted by heating treatment at 30-50 ℃ for 5-30 min, so that uneven crystal nuclei in the fish oil can be destroyed.
Preferably, the organic solvent is one or more of methanol, absolute ethyl alcohol, acetone and n-hexane.
During low-temperature crystallization separation, saturated acids in fish oil fatty acids and low-unsaturation fatty acids such as monoenoic acid and dienoic acid are required to be precipitated in a crystal form, EPA and DHA are still remained in a solution, and the organic solvent still has good solubility for EPA and DHA at a lower temperature, so that EPA and DHA are ensured to be remained in the solution as much as possible.
Preferably, the organic solvent contains 0.8-1.2% by mass of citric acid.
The addition of a small amount of citric acid can promote the rapid crystallization of EPA and DHA and the refinement of crystal grains under the low-temperature condition, and improve the crystallization efficiency.
Preferably, the organic solvent is a mixed solvent of methanol and absolute ethyl alcohol in a volume ratio of (6-12): 1, and the organic solvent further contains 0.9-1.1% by mass of citric acid.
The methanol has the best effect of removing saturated fatty acid, namely the enrichment effect on EPA and DHA, the absolute ethanol has the second best effect, the absolute ethanol has the better effect of removing low unsaturated fatty acid, the ethanol is nontoxic, and the methanol has the advantages of low price, small volatility, high flash point and the like, but has relatively high viscosity and low crystallization temperature. Comprehensively considering, selecting compound methanol and absolute ethyl alcohol as solvents for low-temperature crystallization treatment.
Preferably, the crystallization assistant is one or two of diatomite and argil.
The diatomite and the argil have huge surface areas, and a small amount of the diatomite and the argil are added in the low-temperature crystallization process, so that a large activated surface can be provided for the nucleation of saturated fatty acid in fish oil, the nucleation free energy is reduced, crystals form a compact and ordered structure, and the growth of subsequent crystals on the surface of the crystals is accelerated. And the diatomite and the argil can be removed through centrifugal treatment in the subsequent process, and are safe and non-toxic. The addition amount of the crystallization assistant agent is too small, the number of generated crystal nuclei is small, the crystallization rate is low, and the effect of promoting crystallization is not obvious; if the addition amount is too large, the movement of saturated fatty acid molecules is affected by the existence of too many heterogeneous impurities, so that the diffusion capability of the saturated fatty acid molecules to the crystallization front is weakened, and the crystallization rate is reduced.
Preferably, the low-temperature crystallization is carried out in ultrasonic stirring, wherein the ultrasonic stirring has an ultrasonic power of 300-500 w and a stirring speed n of 15-30 r/min
The fish oil under the supercooling condition is treated by ultrasonic waves, a large number of crystal nuclei are formed due to the flocculation effect of the ultrasonic waves, the cavitation effect of the ultrasonic waves breaks the formed crystal nuclei to form new crystal centers, the secondary nucleation is promoted, the crystallization of saturated fatty acid is accelerated, the crystallization induction time is shortened, the crystal morphology is changed, the crystal particles are fine and uniform, and the hardness is increased. If the stirring force is not enough, local crystal nuclei can be generated, and if the stirring force is too violent, the crystals can be torn, so that the stirring speed is controlled appropriately. The continuous slow stirring is kept in the crystal growing process, which is beneficial to improving the heat transfer efficiency and the growth of crystals.
Preferably, the low-temperature crystallization comprises a first stage and a second stage, wherein the temperature of the first stage is reduced to-10-5 ℃, the temperature is maintained for crystallization for 3-4 hours, the temperature of the second stage is reduced to-30-50 ℃, and the temperature is maintained for crystallization for 2.5-3.5 hours.
The temperature is one of the key factors for the formation and separation of fatty acid crystals, the temperature is too high, heat is difficult to transfer from a solvent and a mixture of fish oil and the mixture, a good supercooling degree cannot be formed, the formation of crystal nuclei is limited, and the non-crystallized liquid after low-temperature crystallization contains a large amount of other components besides EPA and DHA, so that the proportion of EPA and DHA is reduced. Proper crystallization temperature is favorable for the saturated fatty acid to form crystals. However, the crystallization temperature is controlled within the above range by the present invention because the crystallization temperature is too low, EPA and DHA migrate to the crystal nucleus and finally precipitate as crystals, the EPA and DHA enrichment ratio is reduced, and the formed crystals are large and difficult to filter, and the reduction of the temperature increases energy consumption.
The crystallization process is carried out in two steps, the growth of crystal nuclei and the growth of crystals, and once the crystal nuclei are formed, the crystals begin to grow. As long as the system is not saturated, the temperature is not too cold, the fatty acid molecules also have a higher mobility to form crystals, which will continue to form until the system reaches equilibrium. The growth of the crystal requires a certain time, and thus the crystallization time is an important factor. The crystallization time is short, the crystallization is incomplete, some saturated fatty acid components still exist in the organic solvent, and the proportion of EPA and DHA in the final product is reduced; and the crystallization time is too long, which may cause partial EPA and DHA components to tend to crystallize, resulting in the reduction of the content of EPA and DHA. The present invention controls the crystallization time within the above range.
The low-temperature crystallization process is carried out in two stages, when the crystallization temperature is gradually reduced to-10-5 ℃, the crystallization rate of saturated fatty acid in the fish oil is rapidly increased, and when the temperature is reduced again, the crystallization rate is increased and slowed, until the temperature is reduced to-30 to-50 ℃, the increase of the crystallization rate is accelerated again, which shows that the crystallization rate of saturated fatty acid in the fish oil is higher at-10-5 ℃ and-30 to-50 ℃. Therefore, the invention adopts a sectional crystallization method, the first stage of crystallization is carried out at-10 to 5 ℃ to fully crystallize the saturated fatty acid which can be crystallized in the first stage, and then the second stage of crystallization is carried out at-30 to-50 ℃, the first stage of crystallization reduces the pressure of the second stage of crystallization, which is beneficial to reducing energy consumption, and the first stage of crystallization can be used as a crystal nucleus of the second stage of crystallization to be beneficial to the second stage of crystallization.
Preferably, the temperature reduction rate of the first stage is 1-1.5 ℃/min, and the temperature reduction rate of the second stage is 1.3-1.8 ℃/min.
In the separation process of the crystal and the crystallization liquid, the crystal form of the crystal influences the separation effect, and the most stable crystal form is obtained by the cooling rate and the crystallization temperature. The vitreous crystals which cannot be separated are easily formed by rapid cooling with large temperature difference, and the corresponding crystal forms can be obtained only by adopting proper cooling rate to slowly cool to the crystallization temperature. The invention adopts a lower cooling rate in the first stage to facilitate the primary growth of crystals formed by crystal nucleuses, and adopts a faster cooling rate in the second stage to facilitate the rapid formation of the crystals, thereby improving the crystallization efficiency.
Compared with the prior art, the invention has the following beneficial effects: according to the invention, ethyl ester type fish oil is used as a raw material, EPA and DHA are enriched by a low-temperature crystallization method, an organic solvent and a crystallization aid are reasonably selected, technological parameters are optimized, and fish oil with high content of EPA and DHA is obtained by separation and enrichment; the method has the advantages of simple operation, low requirement on instruments and equipment, large single treatment capacity and good separation effect, can better maintain the natural activity of EPA and DHA in a low-temperature environment, and has important reference significance for deep processing and utilization of fish oil and development of polyunsaturated fatty acid series health care products.
Detailed Description
The following are specific examples of the present invention and further describe the technical solutions of the present invention, but the present invention is not limited to these examples.
The method for enriching the fish oil EPA and DHA in the present invention is further explained by the following specific examples 1-12.
Example 1
The method for enriching the fish oil EPA and DHA through low-temperature crystallization comprises the following steps:
heating the ethyl ester type fish oil at 30 ℃ for 30min, wherein the content of EPA and DHA in the ethyl ester type fish oil is 50% by mass;
adding methanol and diatomite into fish oil, wherein the mass ratio of the fish oil to the methanol to the diatomite is 100:200:0.5, and uniformly mixing to form a mixture;
placing the mixture in low-temperature equipment, cooling to 5 ℃ at the speed of 1.0 ℃/min while ultrasonically stirring, performing heat preservation crystallization for 3h, cooling to-30 ℃ at the speed of 1.3 ℃/min, performing heat preservation crystallization for 2.5h, wherein the ultrasonic power is 300w, and the stirring speed n is 15 r/min;
and after low-temperature crystallization, carrying out vacuum filtration to separate crystals and non-crystallized liquid, centrifuging to remove the crystallization assistant in the non-crystallized liquid, and carrying out molecular distillation to remove the organic solvent in the non-crystallized liquid, thereby obtaining the enriched fish oil EPA and DHA. Through detection, the total content of EPA and DHA in the enriched fish oil EPA and DHA is 87.53%.
Example 2
The method for enriching the fish oil EPA and DHA through low-temperature crystallization comprises the following steps:
heating the ethyl ester type fish oil at 35 ℃ for 25min, wherein the content of EPA and DHA in the ethyl ester type fish oil is 55% by mass;
adding absolute ethyl alcohol and argil into fish oil, wherein the mass ratio of the fish oil to the absolute ethyl alcohol to the argil is 100:400:1, and uniformly mixing to form a mixture;
placing the mixture in low-temperature equipment, cooling to 0 ℃ at the speed of 1.3 ℃/min while ultrasonically stirring, performing heat preservation crystallization for 3.5h, cooling to-35 ℃ at the speed of 1.5 ℃/min, performing heat preservation crystallization for 3h, wherein the ultrasonic power is 350w, and the stirring speed n is 20 r/min;
and after low-temperature crystallization, carrying out vacuum filtration to separate crystals and non-crystallized liquid, centrifuging to remove the crystallization assistant in the non-crystallized liquid, and carrying out molecular distillation to remove the organic solvent in the non-crystallized liquid, thereby obtaining the enriched fish oil EPA and DHA. Through detection, the total content of EPA and DHA in the enriched fish oil EPA and DHA is 87.62%.
Example 3
The method for enriching the fish oil EPA and DHA through low-temperature crystallization comprises the following steps:
heating ethyl ester type fish oil at 40 deg.C for 20min, wherein the content of EPA and DHA in the ethyl ester fish oil is 60 wt%;
adding acetone and diatomite into the fish oil, wherein the mass ratio of the fish oil to the acetone to the diatomite is 100:600:1.5, and uniformly mixing to form a mixture;
placing the mixture in low-temperature equipment, cooling to-3 ℃ at the speed of 1.5 ℃/min while ultrasonically stirring, performing heat preservation crystallization for 4h, cooling to-40 ℃ at the speed of 1.8 ℃/min, performing heat preservation crystallization for 3.5h, wherein the ultrasonic power is 400w, and the stirring speed n is 25 r/min;
and after low-temperature crystallization, carrying out vacuum filtration to separate crystals and non-crystallized liquid, centrifuging to remove the crystallization assistant in the non-crystallized liquid, and carrying out molecular distillation to remove the organic solvent in the non-crystallized liquid, thereby obtaining the enriched fish oil EPA and DHA. Through detection, the total content of EPA and DHA in the enriched fish oil EPA and DHA is 88.87%.
Example 4
The method for enriching the fish oil EPA and DHA through low-temperature crystallization comprises the following steps:
heating ethyl ester type fish oil at 40 deg.C for 10min, wherein the content of EPA and DHA in the ethyl ester fish oil is 70 wt%;
adding normal hexane and argil into the fish oil, wherein the mass ratio of the fish oil to the normal hexane to the argil is 100:800:2, and uniformly mixing to form a mixture;
placing the mixture in low-temperature equipment, cooling to-5 ℃ at the speed of 1.4 ℃/min while ultrasonically stirring, performing heat preservation crystallization for 3.5h, cooling to-45 ℃ at the speed of 1.7 ℃/min, performing heat preservation crystallization for 3h, wherein the ultrasonic power is 450w, and the stirring speed n is 30 r/min;
and after low-temperature crystallization, carrying out vacuum filtration to separate crystals and non-crystallized liquid, centrifuging to remove the crystallization assistant in the non-crystallized liquid, and carrying out molecular distillation to remove the organic solvent in the non-crystallized liquid, thereby obtaining the enriched fish oil EPA and DHA. Through detection, the total content of EPA and DHA in the enriched fish oil EPA and DHA is 89.71%.
Example 5
The method for enriching the fish oil EPA and DHA through low-temperature crystallization comprises the following steps:
heating the ethyl ester type fish oil at 50 ℃ for 5min, wherein the content of EPA and DHA in the ethyl ester type fish oil is 65% by mass;
adding an organic solvent and diatomite into fish oil, wherein the mass ratio of the fish oil to the organic solvent to the diatomite is 100:1000:2, uniformly mixing to form a mixture, and the organic solvent is a mixed solvent of methanol and absolute ethanol with the volume ratio of 6: 1;
placing the mixture in low-temperature equipment, cooling to-10 ℃ at the speed of 1.3 ℃/min while ultrasonically stirring, performing heat preservation crystallization for 3.5h, cooling to-50 ℃ at the speed of 1.6 ℃/min, performing heat preservation crystallization for 3h, wherein the ultrasonic power is 500w, and the stirring speed n is 25 r/min;
and after low-temperature crystallization, carrying out vacuum filtration to separate crystals and non-crystallized liquid, centrifuging to remove the crystallization assistant in the non-crystallized liquid, and carrying out molecular distillation to remove the organic solvent in the non-crystallized liquid, thereby obtaining the enriched fish oil EPA and DHA. Through detection, the total content of EPA and DHA in the enriched fish oil EPA and DHA is 89.63%.
Example 6
The method for enriching the fish oil EPA and DHA through low-temperature crystallization comprises the following steps:
heating ethyl ester type fish oil at 40 deg.C for 20min, wherein the content of EPA and DHA in the ethyl ester fish oil is 60 wt%;
adding an organic solvent and a crystallization aid into fish oil, wherein the mass ratio of the fish oil to the organic solvent to the crystallization aid is 100:600:1.5, uniformly mixing to form a mixture, the organic solvent is a mixed solvent of methanol and absolute ethanol with the volume ratio of 9:1, and the crystallization aid is a mixture of diatomite and clay with the mass ratio of 2: 1;
placing the mixture in low-temperature equipment, cooling to-10 ℃ at the speed of 1.3 ℃/min while ultrasonically stirring, performing heat preservation crystallization for 3.5h, cooling to-50 ℃ at the speed of 1.7 ℃/min, performing heat preservation crystallization for 3h, wherein the ultrasonic power is 400w, and the stirring speed n is 20 r/min;
and after low-temperature crystallization, carrying out vacuum filtration to separate crystals and non-crystallized liquid, centrifuging to remove the crystallization assistant in the non-crystallized liquid, and carrying out molecular distillation to remove the organic solvent in the non-crystallized liquid, thereby obtaining the enriched fish oil EPA and DHA. Through detection, the total content of EPA and DHA in the enriched fish oil EPA and DHA is 89.20%.
Example 7
The method for enriching the fish oil EPA and DHA through low-temperature crystallization comprises the following steps:
heating ethyl ester type fish oil at 40 deg.C for 20min, wherein the content of EPA and DHA in the ethyl ester fish oil is 60 wt%;
adding an organic solvent and a crystallization aid into fish oil, wherein the mass ratio of the fish oil to the organic solvent to the crystallization aid is 100:600:1.5, uniformly mixing to form a mixture, the organic solvent is a mixed solvent of methanol and absolute ethanol with the volume ratio of 12:1, and the crystallization aid is a mixture of diatomite and clay with the mass ratio of 1: 1;
placing the mixture in low-temperature equipment, cooling to-10 ℃ at the speed of 1.2 ℃/min while ultrasonically stirring, performing heat preservation crystallization for 3.5h, cooling to-50 ℃ at the speed of 1.6 ℃/min, performing heat preservation crystallization for 3h, wherein the ultrasonic power is 400w, and the stirring speed n is 20 r/min;
and after low-temperature crystallization, carrying out vacuum filtration to separate crystals and non-crystallized liquid, centrifuging to remove the crystallization assistant in the non-crystallized liquid, and carrying out molecular distillation to remove the organic solvent in the non-crystallized liquid, thereby obtaining the enriched fish oil EPA and DHA. Through detection, the total content of EPA and DHA in the enriched fish oil EPA and DHA is 89.21%.
Example 8
The method for enriching EPA and DHA in fish oil through low-temperature crystallization comprises the following steps, wherein the EPA and DHA content in the ethyl esterification fish oil is 60% by mass:
heating ethyl ester type fish oil at 40 deg.C for 20 min;
adding an organic solvent and a crystallization aid into fish oil, wherein the mass ratio of the fish oil to the organic solvent to the crystallization aid is 100:600:1.5, uniformly mixing to form a mixture, the organic solvent is a mixed solvent of methanol and absolute ethanol with the volume ratio of 9:1, the organic solvent contains 0.8% of citric acid by mass, and the crystallization aid is a mixture of diatomite and argil with the mass ratio of 2: 1;
placing the mixture in low-temperature equipment, cooling to-10 ℃ at the speed of 1.2 ℃/min while ultrasonically stirring, performing heat preservation crystallization for 3.5h, cooling to-50 ℃ at the speed of 1.6 ℃/min, performing heat preservation crystallization for 3h, wherein the ultrasonic power is 400w, and the stirring speed n is 20 r/min;
and after low-temperature crystallization, carrying out vacuum filtration to separate crystals and non-crystallized liquid, centrifuging to remove the crystallization assistant in the non-crystallized liquid, and carrying out molecular distillation to remove the organic solvent in the non-crystallized liquid, thereby obtaining the enriched fish oil EPA and DHA. Through detection, the total content of EPA and DHA in the enriched fish oil EPA and DHA is 90.02%.
Example 9
The method for enriching EPA and DHA in fish oil through low-temperature crystallization comprises the following steps, wherein the EPA and DHA content in the ethyl esterification fish oil is 60% by mass:
heating ethyl ester type fish oil at 40 deg.C for 20 min;
adding an organic solvent and a crystallization aid into fish oil, wherein the mass ratio of the fish oil to the organic solvent to the crystallization aid is 100:600:1.5, uniformly mixing to form a mixture, the organic solvent is a mixed solvent of methanol and absolute ethanol with the volume ratio of 9:1, the organic solvent contains 0.9% of citric acid by mass, and the crystallization aid is a mixture of diatomite and argil with the mass ratio of 2: 1;
placing the mixture in low-temperature equipment, cooling to-10 ℃ at the speed of 1.3 ℃/min while ultrasonically stirring, performing heat preservation crystallization for 3.5h, cooling to-50 ℃ at the speed of 1.8 ℃/min, performing heat preservation crystallization for 3h, wherein the ultrasonic power is 400w, and the stirring speed n is 20 r/min;
and after low-temperature crystallization, carrying out vacuum filtration to separate crystals and non-crystallized liquid, centrifuging to remove the crystallization assistant in the non-crystallized liquid, and carrying out molecular distillation to remove the organic solvent in the non-crystallized liquid, thereby obtaining the enriched fish oil EPA and DHA. Through detection, the total content of EPA and DHA in the enriched fish oil EPA and DHA is 90.11%.
Example 10
The method for enriching EPA and DHA in fish oil through low-temperature crystallization comprises the following steps, wherein the EPA and DHA content in the ethyl esterification fish oil is 60% by mass:
heating ethyl ester type fish oil at 40 deg.C for 20 min;
adding an organic solvent and a crystallization aid into fish oil, wherein the mass ratio of the fish oil to the organic solvent to the crystallization aid is 100:600:1.5, uniformly mixing to form a mixture, the organic solvent is a mixed solvent of methanol and absolute ethanol with the volume ratio of 9:1, the organic solvent contains 1.0% by mass of citric acid, and the crystallization aid is a mixture of diatomite and clay with the mass ratio of 2: 1;
placing the mixture in low-temperature equipment, cooling to-10 ℃ at the speed of 1.1 ℃/min while ultrasonically stirring, performing heat preservation crystallization for 3.5h, cooling to-50 ℃ at the speed of 1.6 ℃/min, performing heat preservation crystallization for 3h, wherein the ultrasonic power is 400w, and the stirring speed n is 20 r/min;
and after low-temperature crystallization, carrying out vacuum filtration to separate crystals and non-crystallized liquid, centrifuging to remove the crystallization assistant in the non-crystallized liquid, and carrying out molecular distillation to remove the organic solvent in the non-crystallized liquid, thereby obtaining the enriched fish oil EPA and DHA. Through detection, the total content of EPA and DHA in the enriched fish oil EPA and DHA is 92.00%.
Example 11
The method for enriching EPA and DHA in fish oil through low-temperature crystallization comprises the following steps, wherein the EPA and DHA content in the ethyl esterification fish oil is 60% by mass:
heating ethyl ester type fish oil at 40 deg.C for 20 min;
adding an organic solvent and a crystallization aid into fish oil, wherein the mass ratio of the fish oil to the organic solvent to the crystallization aid is 100:600:1.5, uniformly mixing to form a mixture, the organic solvent is a mixed solvent of methanol and absolute ethanol with the volume ratio of 9:1, the organic solvent contains 1.1% by mass of citric acid, and the crystallization aid is a mixture of diatomite and clay with the mass ratio of 2: 1;
placing the mixture in low-temperature equipment, cooling to-10 ℃ at the speed of 1.3 ℃/min while ultrasonically stirring, performing heat preservation crystallization for 3.5h, cooling to-50 ℃ at the speed of 1.7 ℃/min, performing heat preservation crystallization for 3h, wherein the ultrasonic power is 400w, and the stirring speed n is 20 r/min;
and after low-temperature crystallization, carrying out vacuum filtration to separate crystals and non-crystallized liquid, centrifuging to remove the crystallization assistant in the non-crystallized liquid, and carrying out molecular distillation to remove the organic solvent in the non-crystallized liquid, thereby obtaining the enriched fish oil EPA and DHA. Through detection, the total content of EPA and DHA in the enriched fish oil EPA and DHA is 91.65%.
Example 12
The method for enriching EPA and DHA in fish oil through low-temperature crystallization comprises the following steps, wherein the EPA and DHA content in the ethyl esterification fish oil is 60% by mass:
heating ethyl ester type fish oil at 40 deg.C for 20 min;
adding an organic solvent and a crystallization aid into fish oil, wherein the mass ratio of the fish oil to the organic solvent to the crystallization aid is 100:600:1.5, uniformly mixing to form a mixture, the organic solvent is a mixed solvent of methanol and absolute ethanol with the volume ratio of 9:1, the organic solvent contains 1.2% of citric acid by mass, and the crystallization aid is a mixture of diatomite and argil with the mass ratio of 2: 1;
placing the mixture in low-temperature equipment, cooling to-10 ℃ at the speed of 1.2 ℃/min while ultrasonically stirring, performing heat preservation crystallization for 3.5h, cooling to-50 ℃ at the speed of 1.5 ℃/min, performing heat preservation crystallization for 3h, wherein the ultrasonic power is 400w, and the stirring speed n is 20 r/min;
and after low-temperature crystallization, carrying out vacuum filtration to separate crystals and non-crystallized liquid, centrifuging to remove the crystallization assistant in the non-crystallized liquid, and carrying out molecular distillation to remove the organic solvent in the non-crystallized liquid, thereby obtaining the enriched fish oil EPA and DHA. Through detection, the total content of EPA and DHA in the enriched fish oil EPA and DHA is 91.86%.
Comparative example 1
No crystallization promoter was added during the enrichment process, and the rest was the same as in example 10. Through detection, the total content of EPA and DHA in the enriched fish oil EPA and DHA is 83.16%.
Comparative example 2
In the low-temperature freezing process, the temperature is reduced to minus 30 ℃ at the speed of 18 ℃/min while ultrasonic stirring, and the heat preservation crystallization is carried out for 5.5h, and the rest is the same as that of the embodiment 10. Through detection, the total content of EPA and DHA in the enriched fish oil EPA and DHA is 83.93%.
Comparative example 3
In the low-temperature freezing process, ultrasonic waves were not used, and the procedure was otherwise the same as in example 10. Through detection, the total content of EPA and DHA in the enriched fish oil EPA and DHA is 86.19%.
In conclusion, the ethyl ester type fish oil is used as a raw material, EPA and DHA are enriched by a low-temperature crystallization method, and technological parameters are reasonably optimized in the enrichment process; diatomite and clay are added as crystallization promoting agents, the proportion of heterogeneous nucleation is increased, the supercooling degree is reduced, and the crystallization temperature is increased; ultrasonic stirring is assisted, so that the crystallization efficiency is increased; a two-section crystallization method is adopted in the crystallization process, so that the crystallization energy consumption is reduced, and the crystallization efficiency is increased; the method is simple and convenient to operate, has low requirements on instruments and equipment, large single treatment capacity and good separation effect, and can better maintain the natural activity of EPA and DHA in a low-temperature environment.
The specific embodiments described herein are merely illustrative of the spirit of the invention. Various modifications or additions may be made to the described embodiments or alternatives may be employed by those skilled in the art without departing from the spirit or ambit of the invention as defined in the appended claims.
Claims (5)
1. A method for enriching EPA and DHA in fish oil through low-temperature crystallization is characterized by comprising the following steps:
pretreating fish oil, adding an organic solvent and a crystallization aid into the fish oil, wherein the mass ratio of the fish oil to the organic solvent to the crystallization aid is 100 (200-1000) to 0.5-2, uniformly mixing, performing low-temperature crystallization, performing vacuum filtration to separate crystals and non-crystallized liquid, and removing the organic solvent and the crystallization aid from the non-crystallized liquid to obtain enriched fish oil EPA and DHA;
the fish oil is ethyl ester type fish oil,
the pretreatment is to heat the fish oil at 30-50 ℃ for 5-30 min,
the organic solvent is one or more of methanol, absolute ethyl alcohol, acetone and normal hexane, the organic solvent contains 0.8-1.2% by mass of citric acid,
the crystallization aid is one or two of diatomite and argil.
2. The method for low-temperature crystallization enrichment of fish oil EPA and DHA as claimed in claim 1, wherein the organic solvent is a mixed solvent of methanol and absolute ethanol with a volume ratio of (6-12): 1, and the organic solvent further contains 0.9-1.1% by mass of citric acid.
3. The method for enriching fish oil EPA and DHA through low-temperature crystallization according to claim 1, wherein the low-temperature crystallization is performed in ultrasonic stirring, the ultrasonic power of the ultrasonic stirring is 300-500 w, and the stirring speed n = 15-30 r/min.
4. The method for enriching the EPA and the DHA in the fish oil through low-temperature crystallization according to claim 1, wherein the low-temperature crystallization comprises a first stage and a second stage, the temperature of the first stage is reduced to-10-5 ℃, the temperature is maintained for crystallization for 3-4 h, the temperature of the second stage is reduced to-30-50 ℃, and the temperature is maintained for crystallization for 2.5-3.5 h.
5. The method for low-temperature crystallization enrichment of fish oil EPA and DHA as claimed in claim 4, wherein the temperature reduction rate of the first stage is 1.0-1.5 ℃/min, and the temperature reduction rate of the second stage is 1.3-1.8 ℃/min.
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