KR101024491B1 - The Method of Extracting Lipids from Marine Animals - Google Patents

Abstract

The present invention comprises the steps of (a) heating 10-60 (v / v)% ethanol to a temperature of 40-80 ℃; (b) removing oxygen from the ethanol using nitrogen; And (c) extracting the ethanol from which oxygen is removed in step (b) by spraying the marine animal pulverized at a pressure of 50-3,000 bar and extracting for 2-5 hours. Relates to marine animal extracts.

Marine Animal, Krill Shrimp, High Pressure Pump, Astaxanthin, DHA

Description

 The Method of Extracting Lipids from Marine Animals

The present invention relates to marine animal extracts, and more particularly, to a marine animal extract enhanced with lenolenic acid (Dinolenic acid), DHA (Docosahexaenoic acid), EPA (Eicosapentaenoic acid), Phospholipid (Phospholipid) and Astaxanthin (Astaxanthin) It is about.

As the demand for omega-3 fatty acids increases with increasing income and health concern, there is a need for an efficient omega-3 fatty acid extraction process. Many are destroyed. In addition, krill oil is known to have a much higher absorption effect on the human body than fish oil because its structure is similar to the phospholipids that make up the human fat cell wall. Krill oil contains vitamins A, E, and D in addition to lipids such as omega-3, and contains astaxanthin, a particularly powerful antioxidant, and it has been reported that krill oil's antioxidant activity is 48 times that of normal fish oil. .

Krill oil is known to have the following benefits and uses:

(1) Medical use

Krill oil and its fractions have anti-inflammatory properties and are known to help alleviate the symptoms of cardiovascular disease. It is also known to inhibit the development of kidney disease.

(2) dietary regulators

Krill oil can also be used as a dietary source of dietary regulation of the human body. These fatty acids are essential for the proper development of the brain and eyes and are rich in fat-soluble vitamins A, D and E and carotenoids.

(3) cosmetics

Krill oil is highly moisturizing and is used for moisturizing creams.

(4) fish farming

In krill oil, there are high concentrations of fatty acid linolenic acid, DHA and EPA. These fatty acids are essential nutrients and useful as a feed for fish. In particular, krill oil is rich in astaxanthin and can be used as a feed for red meats.

(5) animal feed

Animal foods rich in omega-3 fatty acids can reduce cholesterol levels in meat and increase levels of unsaturated fatty acids. This property is already used in the poultry industry to improve egg quality.

On the other hand, various methods of extracting krill oil are known. Generally, a method of extracting oil using organic solvents such as hexane and ethanol is known. In addition, methods using solvents such as acetone, methanol or chloroform are known. As the extraction method, a reduced pressure distillation method, a low temperature extraction method or a supercritical extraction method is known.

U.S. Patent No. 4,331,695 discloses the use of solvents that are gaseous at room temperature, such as propane, butane or hexane. This extraction method is carried out at a temperature of 15 to 80 ° C. and the extracted oil is extracted and separated under high temperature and high pressure conditions of 50 to 200 ° C. Hexane, however, is an undesirable solvent for the extraction of marine animals such as krill. Moreover, the high temperatures used in the extraction separation step are likely to deform the extracted oil.

Canadian Patent Application No. 2,115,571 discloses oil extraction methods of various brown and red algae. This method provides, for example, Soxhlet extraction using nearly pure ethanol for 40 hours.

U. S. Patent No. 5,006, 281 discloses a method for extracting oil from marine and aquatic animals such as fish. The marine and aquatic animals are first treated with antioxidants, finely divided and centrifuged to separate the oil phase from the water and solid phases. The oil phase is then treated with an antioxidant to remove undesirable flavors or flavors.

Canadian Patent Application No. 1,098,900 discloses a method for extracting oil from krill. This method involves emulsifying fresh or thawed krill in an aqueous medium. The oil fraction is recovered by centrifugation.

J. Biol . In a paper published in Chem ., 226: 497-509 (1957), “A simple method for the isolation and purification of total lipids from animal tissues,” Folk presented an extraction method using chloroform and methanol. This method cannot be used for oil extraction for food or feed due to the toxicity of the solvent used.

As such, the prior art processes usually consist of a process that extracts only one substance, as well as not being practically available or low in yield. Korean Patent Publication No. 2002-0084371 discloses a method for preparing phospholipids containing DHA and EPA, which are the main components of omega-3, but this method is not suitable as an oil extraction method for food use due to the toxicity of solvents, as well as pigment extraction. Not suitable for

The present inventors provide marine animal extracts enriched with linolenic acid, DHA, EPA, phospholipids and astaxanthin pigments in high yield without chemical and thermal denaturation of the active substances from marine animals.

According to one aspect of the invention, the invention comprises the steps of (a) heating 10-60 (v / v)% ethanol to a temperature of 40-80 ℃; (b) removing oxygen from the ethanol using nitrogen; And (c) extracting the ethanol from which oxygen is removed in step (b) by spraying the marine animal pulverized at a pressure of 50-3,000 bar and extracting for 2-5 hours. Provide marine animal extracts.

The present inventors have endeavored to extract marine animal extracts from marine animals in high yield without chemical and thermal denaturation of the active substances. As a result, marine animal extracts extracted under constant extraction conditions (temperature, pressure, etc.) using a mixed solvent of alcohol and water are used to enhance linolenic acid, DHA, EPA, phospholipids, and astaxanthin pigments, and chemical and thermal effects. It was confirmed that the extraction in high yield without denaturation.

Marine animals used in the present invention include crustaceans such as shrimp or crab, arthropods, and salmon, sea bream, carp or goldfish containing omega-3 fatty acids (linolenic acid, DHA and EPA), phospholipids and astaxanthin pigments, and the like. Various marine animals, such as fish, are included, Preferably krill, shrimp, crab, salmon, or sea bream, More preferably, krill.

Hereinafter, an extraction method for extracting the marine animal extract of the present invention will be described in detail with reference to the accompanying drawings as follows:

(a) heating the solvent

First, 10-60 (v / v)% ethanol is heated to a temperature of 40-80 ° C.

According to a preferred embodiment of the present invention, the ethanol used in the present invention may be 30-50 (v / v)%. In case of less than 10 (v / v)% ethanol, it takes long time to elute fat-soluble substance and very low extraction efficiency.In case of more than 60 (v / v)% ethanol, extraction efficiency is improved but As flammability increases, there is a need for facilities such as explosion-proof facilities in preparation for an accident such as fire and explosion, which increases the facility investment and management burden.

According to another preferred embodiment of the invention, the ethanol used in the invention has a temperature of 45-55 ° C. If the temperature of ethanol is less than 40 ℃, the ethanol penetrates into the extract longer, so it takes a lot of extraction time and has low extraction efficiency.When it exceeds 80 ℃, the extraction efficiency increases but the temperature of the extract increases. It causes heat denaturation and has a disadvantage in that a lot of ethanol loss occurs because it exceeds the boiling point of ethanol.

(b) removing oxygen from the ethanol using nitrogen

According to the extraction method used in the present invention, since the oxygen contained in the ethanol of step (a) promotes oxidation of the extract, oxygen is removed using nitrogen generated through the nitrogen generator.

(c) spraying the oxygen-depleted ethanol to the ground animal

In step (b), the oxygen-free ethanol is sprayed to the marine animal crushed at a pressure of 50-3,000 bar and extracted for 2-5 hours.

Marine animals used in the present invention contain a large amount of fat-soluble vitamins (A, E and D) and antioxidant astaxanthin in addition to lipids such as omega-3.

On the other hand, marine animals used in the present invention are not significantly affected by conditions (freezing or thawing) or drying conditions (nature, hot air, vacuum or freezing, etc.) and can be used by grinding to a certain size to increase the surface area.

According to a preferred embodiment of the present invention, the pressure may be 200-600 bar, more preferably 350-450 bar. If the extraction pressure is less than 50 bar, the discharge pressure is low to not sufficiently agitate the extracts, and ethanol does not penetrate sufficiently to the inside of the extract, resulting in a poor extraction efficiency. Although very good, there is a disadvantage in that the installation cost increases due to the extraction efficiency.

The extraction method for extracting the marine animal extract of the present invention may further include the following step (d).

(d) centrifugation and vacuum distillation of the resultant (c)

In addition, the extraction method is to recover the marine animal extract containing water-containing ethanol by centrifuging the resultant (c) and distilled it under reduced pressure at 25-60 ℃ to evaporate ethanol and water marine animals containing the active substance Extracts can be obtained. The reason for distillation under reduced pressure at 25-60 ° C. is to safely separate many heat-sensitive antioxidants and active substances by performing the separation process at the above temperature.

According to another preferred embodiment of the present invention, the marine animal extract of the present invention may be fortified with linolenic acid, DHA, EPA, phospholipids and astaxanthin.

As described in detail above, the present invention is a marine animal extract extracted under a constant extraction conditions (temperature, pressure, etc.) using a mixed solvent of alcohol and water, omega-3 (linolenic acid, DHA, EPA without chemical thermal denaturation) ), Phospholipids and astaxanthin pigments. In addition, in the present invention by extracting using a solvent harmless to the human body, the marine animal extract of the present invention can be applied as a pharmaceutical, cosmetic or food composition, the by-product can be used as animal feed.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, and the scope of the present invention is not limited by these examples in accordance with the gist of the present invention to those skilled in the art. Will be self explanatory.

Example

Example  One:

100 kg of crushed krill shrimp were filled in a reactor having an internal volume of 200 L, the reactor was closed, and 30-60 (v / v)% ethanol heated using a high pressure pump was injected into the reactor through a nozzle attached to the reactor. Pigment and lipid were extracted.

At this time, the extraction temperature and pressure were respectively adjusted to the extraction temperature of 45-55 ℃ using a heater attached to the outside of the extraction tank and the high pressure pump mounted to the front of the extraction tank and extracted by changing the pressure to 350-400bar. The total extraction time was 2-3 hours. The krill shrimp obtained after extraction was transferred to a continuous centrifuge from the transfer pump to separate the solvent, and the solvent was concentrated under reduced pressure to recover the pigment and lipid. The weight of the recovered product was measured to compare the extraction rate and the extraction yield obtained in the comparative example.

The composition of the dried krill shrimp used in the present invention is shown in Table 1 below:

Furtherance % Per 100g Fresh krill Dry krill moisture 79.9  4.3 Crude fat  4.3 25.7 Crude protein 12.6 62.2 Etc  3.2  7.8

And the fatty acid composition of the krill shrimp extracted by the above method is summarized in Table 2, and the phospholipid and astaxanthin content in Table 3.

Comparative example  One

Ethyl ether was used as a solvent, and the extraction method was the soxhlet extraction method described in Canadian Patent Application No. 2,115,571, and krill was extracted by Soxhlet extraction method.

Fatty acid composition of krill shrimp extract extracted by Soxhlet extraction method using ethyl ether is summarized in Table 2 and phospholipid and astaxanthin content in Table 3.

Comparative example  2

Ethyl ether was used as a solvent, and the extraction method was the soxhlet extraction method described in Canadian Patent Application No. 2,115,571, and krill was extracted by Soxhlet extraction method.

Fatty acid composition of krill shrimp extract extracted by Soxhlet extract using ethanol is shown in Table 2 and phospholipids and astaxanthin contents are shown in Table 3.

Krill  Fatty acid composition, phospholipids and Astaxanthin  content

Fatty acid analysis gas-liquid chromatograms of krill shrimp extracts of Example 1, Comparative Example 1 and Comparative Example 2 are shown in FIGS.

Analytical conditions of the gas-liquid chromatograph are shown in Table 2 below:

Column HP-FFAP, 25 m, 0.32 mm, 0.5 Carrier Hellium 46.6 psi Oven 150 ° C (1 min), to 230 ° C (2.9 ° C / min), 230 ° C (1 min) Split Ratio 20/1 Injector Temp. 260 ℃ Injection Size 1.5 Detector Temp. 260 ℃

As shown in Table 2 above, the column used for analysis was HP-FFAP (25 m, 0.32 mm, 0.5), and the oven temperature was initially raised to 150 ° C. and then raised to 230 ° C. at 2.9 ° C./min to 230 When the temperature was reached, the measurement was completed after 1 minute. At this time, the temperature of the sample inlet was 260 ° C, the detector temperature was 260 ° C, and the detector used FID.

The fatty acid composition of the krill shrimp extract of Example 1, Comparative Example 1 and Comparative Example 2 is shown in Table 3 below:

Item Example Comparative Example 1 Comparative Example 2 Extraction yield 14.54 19.74 10.73 lauric acid C12: 0 0.31 0.36 0.48 myristic acid C 15.29 17.47 22.44 myristoleic acid C14: 1 0.34 0.22 0.21 palmitic acid C16: 0 22.60 24.29 23.94 palmitoleic acid C16: 1 1.27 1.07 1.05 stearic acid C18: 0 1.80 1.75 1.67 oleic acid C18: n9c 17.80 13.88 13.17 linoleic acid C18: 2n6c 3.07 2.72 2.87 linolenic acid C18: 3n3 2.33 1.21 1.17 cis-11-eicoseneic acid C20: 1 1.04 0.92 0.84 cis-11,14-eicosadienonic acid C20: 2 2.30 2.08 1.92 EPA (cis-5,8,11,14,17-eicosapentaenoic acid C20: 5n-3) 14.21 10.93 13.36 DHA (cis-4, 7, 10, 13,16, 19-docosahexaenoic acid C22: 6n-3) 8.53 5.22 4.11 unknown 1 12.42 9.33 8.38 unknown 2 7.38 7.32 7.72 Sum 100.0 100.0 100.0

As can be seen in Table 3, the yield of the final krill shrimp extract can be seen that Example 1 is 14.84%, Comparative Example 1 is about 19.74% and Comparative Example 2 is about 10.73%. The extraction yield was highest in Comparative Example 1, then Example 1 was higher, and Comparative Example 2 was the lowest.

In addition, in the case of linolenic acid among the omega-3 fatty acid components required by the present invention, Example 1 was the highest, Comparative Examples 1 and 2 were similar, Example 1 was the highest in EPA, and Comparative Example 2 was then It was high and Comparative Example 1 was the lowest. On the other hand, in the case of DHA, it was found that Example 1 was significantly higher, Comparative Example 1 was higher, and Comparative Example 2 was the lowest.

In addition, since the ethyl ether which is the organic solvent used in the comparative example 1 is not acceptable as a solvent in a food method, the comparative example 1 is hard to use practically.

Phospholipids and astaxanthin content of the krill shrimp extract of Example 1, Comparative Example 1 and Comparative Example 2 are shown in Table 4. The phospholipid content was measured by Baginski method, and the astaxanthin content was analyzed by liquid chromatography.

- Phospholipids (%) Astaxanthin (ppm) Example 1 54.2 126 Comparative Example 1 8.5 104 Comparative Example 2 39.7 59

As can be seen in Table 4, it was found that the krill shrimp extract of Example 1 contains the most phospholipids and astaxanthin. In phospholipids, Comparative Example 2 was the second highest and Comparative Example 1 was the lowest. In the case of astaxanthin, Comparative Example 1 was the highest after Example 1, and Comparative Example 2 was the lowest.

Therefore, it can be confirmed that phospholipids and astaxanthin are most strengthened in the krill shrimp extract according to Example 1.

Example  2:

To prepare a krill shrimp extract in the same manner as in Example 1, by varying the extraction pressure phospholipid extraction efficiency is summarized as shown in Table 5 below:

Phospholipid Extraction Efficiency by Pressure (Temperature: 50 ℃) Pressure (bar) 50 100 500 1000 2000 3000 Extraction rate (%) 43.68 43.87 48.38 51.92 51.35 54.25

As can be seen in Table 5, it can be seen that the maximum recovery of phospholipids in the final krill shrimp extract is about 54.25%.

Example  3:

To prepare a krill shrimp extract in the same manner as in Example 1, by varying the content of water contained in ethanol astaxanthin pigment extraction efficiency is summarized in Table 6 below:

Astaxanthin pigment extraction efficiency (temperature: 50 ℃, pressure: 1000 bar) Water-EtOH (V / V)% 0 10 20 30 40 50 Extraction rate (ppm) 13 45 83 110 115 126

As can be seen in Table 6, the maximum recovery of the astaxanthin pigment in the final krill shrimp extract was about 126 ppm.

As a result of experiments with different extraction conditions in Examples 2, 3 and 4, the optimum separation conditions of krill shrimp extract was derived.

In addition, through Examples 2, 3 and 4, the extraction efficiency of the krill shrimp extract of the present invention was the maximum, it was confirmed that the conditions of the highest omega-3 content.

In preparing the krill shrimp extract of the present invention, it was found that the extraction time should be within 5 hours while maintaining the heating temperature at 80 ° C. or less in order to prevent the denaturation of DHA and the like in using water-containing ethanol.

Detailed description of the extraction process of the marine animal extract of the present invention as follows:

The pulverized krill was introduced into the reactor, heated through a nozzle attached to the reactor, and 10-60 (v / v)% ethanol, in which oxygen was removed by nitrogen gas, was continuously supplied to the reactor using a high pressure pump. Perform the extraction process. When the mixture of krill and 10-60 (v / v)% ethanol becomes a certain volume or more in the reactor, it is sent to the continuous centrifuge through the solvent recovery tube installed in the reactor, and the solvent and the precipitate are separated and fed back into the reactor. At this time, the extraction conditions of the reaction tank may be performed for 2-5 hours at a temperature of 40-80 ℃, pressure 50-3,000 bar. Water-containing ethanol continuously introduced into the extractor is stirred with krill filled in the reaction tank to extract the pigment and oil containing the active substance.

Then, after the extraction process, the contents of the reaction tank is transferred to a centrifuge using a transfer pump to recover water-containing ethanol through centrifugation. The recovered water-containing ethanol was distilled under reduced pressure at 25-60 ° C. to evaporate ethanol and water to recover the lipid component containing the pigment. The water-containing ethanol recovered by distillation under reduced pressure is reused while adjusting the concentration and storing it in a solvent tank.

As described above in detail specific parts of the present invention, for those skilled in the art, these specific techniques are merely preferred embodiments, and the scope of the present invention is not limited thereto.

1 shows a fatty acid analysis gas-liquid chromatogram of krill shrimp extract according to Example 1. FIG.

2 shows a fatty acid analysis gas-liquid chromatogram of krill shrimp extract according to Comparative Example 1.

3 shows a fatty acid analysis gas-liquid chromatogram of krill shrimp extract according to Comparative Example 2.

Claims (7)

(a) heating 30-60 (v / v)% ethanol to a temperature of 45-80 ° C .; (b) removing oxygen from the ethanol using nitrogen; And (c) extracting the lipid from the marine animal, wherein the oxygen-derived ethanol is sprayed on the marine animal pulverized at a pressure of 50-3,000 bar for 2-5 hours in step (b). How to extract. delete delete The method of claim 1, wherein the pressure in step (c) is 200-600 bar. The method of claim 1, wherein the extraction method further comprises the step of (d) after step (c) (d) centrifuging the resultant (c) and distilling under reduced pressure at 25-60 ° C to evaporate ethanol and water. A method of extracting lipids from marine animals. The method of claim 1, wherein the marine animal is selected from the group consisting of krill, shrimp, crab, salmon, and sea bream. delete

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