CN113061482A - A kind of method for extracting earthworm active crude fat - Google Patents
A kind of method for extracting earthworm active crude fat Download PDFInfo
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- CN113061482A CN113061482A CN202110365302.9A CN202110365302A CN113061482A CN 113061482 A CN113061482 A CN 113061482A CN 202110365302 A CN202110365302 A CN 202110365302A CN 113061482 A CN113061482 A CN 113061482A
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- 241000361919 Metaphire sieboldi Species 0.000 title abstract description 86
- 238000000034 method Methods 0.000 title abstract description 28
- 235000019784 crude fat Nutrition 0.000 title abstract description 24
- 241001233061 earthworms Species 0.000 abstract description 62
- 235000019197 fats Nutrition 0.000 abstract description 48
- 238000000605 extraction Methods 0.000 abstract description 19
- 238000004108 freeze drying Methods 0.000 abstract description 9
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- 239000003925 fat Substances 0.000 abstract 4
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- 238000002360 preparation method Methods 0.000 description 10
- 239000012535 impurity Substances 0.000 description 9
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- 239000003795 chemical substances by application Substances 0.000 description 5
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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
- C11B1/025—Pretreatment by enzymes or microorganisms, living or dead
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
本发明涉及一种提取蚯蚓活性粗脂肪的方法，包括以下步骤：（1）蚯蚓清肠；（2）蚯蚓自溶酶解；（3）蚯蚓冻干；（4）蚯蚓脂肪提取。通过本发明，采用“酶解+低温干燥+有机萃取”的技术方法，提取蚯蚓体中的活性脂肪，为后续蚯蚓脂肪产品的深加工提供原料。目前，市场对蚯蚓的开发利用主要集中在活性酶，忽视对脂肪的开发利用。本专利市场实施可能性大，预期经济效益显著。
The invention relates to a method for extracting active crude fat from earthworms, comprising the following steps: (1) intestinal cleansing of earthworms; (2) autolytic enzymolysis of earthworms; (3) freeze-drying of earthworms; and (4) extraction of earthworm fat. Through the present invention, the technical method of "enzymatic hydrolysis + low temperature drying + organic extraction" is adopted to extract the active fat in the earthworm body and provide raw materials for the subsequent deep processing of the earthworm fat product. At present, the development and utilization of earthworms in the market mainly focus on active enzymes, ignoring the development and utilization of fats. The patent market has a high possibility of implementation, and the expected economic benefits are significant.
Description
Technical Field
The invention relates to a method for extracting earthworm active crude fat, belonging to the field of applied biotechnology.
Background
Earthworms, also known as earthworms, are terrestrial invertebrates of the oligochaeta of Annelida, whose life processes can significantly affect soil fertility, and are called "ecosystem engineers". The earthworms are high in feeding impurity and reproduction efficiency, play a special role in aspects of material circulation, biodiversity and the like, so that the earthworms are used for treating high-water-content organic solid wastes such as domestic garbage, livestock and poultry manure and the like to develop an important environmental protection industry, and how to deeply develop the earthworms and utilize high added values becomes a focus of attention at home and abroad. Currently, the deep processing of earthworms mainly focuses on the liquefaction of earthworms and the extraction of high-activity enzyme preparations from earthworm bodies, and the like, and the systematic research on the application of fat in the earthworm bodies is lacked.
The fat of earthworm body is mainly distributed in the body wall and digestive tract of earthworm and participates in synthesizing fat body, glycogen and cell membrane structure, and the components of fatty acid, sterol and the like in the earthworm body play an important role in the growth, development and reproduction of the earthworm. Researches show that the crude fat content of different earthworm living bodies is about 1.5-4.5%. The earthworm fat is efficiently extracted while the biological activity of the earthworm fat is maintained, so that the utilization value of earthworms is forcefully improved, and the healthy development of the earthworm industry is forcefully promoted.
Disclosure of Invention
The present invention has been made in view of the above problems, and an object of the present invention is to provide a method for extracting earthworm-active crude fat.
The technical scheme of the invention is as follows: a method for extracting earthworm active crude fat is characterized by comprising the following steps:
(1) clearing intestines by earthworms; putting fresh earthworms into tap water, and enabling the earthworms to spit out materials swallowed into cavities and intestines in the tap water, wherein the amount of the tap water is 1-2 times of the weight of the earthworms; standing for 6-8 hours, fishing out the earthworms, cleaning with flowing tap water, and removing impurities to obtain the earthworms with cleared intestines for later use;
(2) carrying out autolysis enzymolysis on earthworms; putting the earthworms with the cleared intestines into a heat-preserving barrel, and uniformly adding a sugar preparation, wherein the adding amount of the sugar preparation is 2-4% of the weight of the earthworms with the cleared intestines; forcibly stirring the mixture of the earthworms and the sugar preparation at the stirring speed of 5 revolutions per minute, keeping the temperature of a heat-preserving container at 45-55 ℃, filtering the mixture by using a 5 mm screen to remove impurities after the earthworms are subjected to enzymolysis for 12-24 hours, and collecting an earthworm enzymolysis solution for later use;
(3) freeze-drying earthworms; putting the earthworm enzymolysis liquid into a vacuum freeze drying box, and freeze drying to obtain earthworm freeze-dried powder;
(4) fat extraction; and extracting fat in the earthworm freeze-dried powder by a Soxhlet extraction method.
In the step (2), the sugar preparation comprises the following components: glucose: fructose, sucrose: glucose: the weight ratio of fructose is 3-5: 1-3: 2 to 4.
In the step (3), the water content of the freeze-dried powder is required to be lower than 2%.
In the step (3), the fat in the earthworm freeze-dried powder is extracted by a Soxhlet extraction method, which comprises the following steps:
step 1), preparing petroleum ether with a low boiling range of 30-60 ℃ as an extracting agent; putting the freeze-dried earthworm powder into a filter paper bag, sealing, and putting into an extraction tube; pouring 1/2 volumes of the extracting agent into a fat bottle of a Soxhlet extractor, and connecting the Soxhlet extractor;
step 2), heating the fat bottle at a constant temperature of 45-55 ℃, evaporating and gasifying an extractant in the fat bottle, feeding the extractant into a condensing tube, condensing the extractant, dripping the condensed extractant into the extracting tube to extract fat in the earthworm freeze-dried powder, and refluxing the extractant into the fat bottle when the upper liquid level of the extractant in the extracting tube is higher than a reflux tube;
step 3), carrying out reciprocating reflux extraction for 4 hours according to the step 2) to obtain a mixture of crude earthworm fat and petroleum ether;
step 4), transferring the mixture to a rotary evaporator, and recovering pure petroleum ether at constant temperature of 45-55 ℃ to obtain a concentrated earthworm crude fat and petroleum ether mixture;
step 5), placing the concentrated mixture of the crude earthworm fat and the petroleum ether in a fume hood, and exhausting and volatilizing for 12 hours to fully volatilize the residual petroleum ether to obtain the crude earthworm fat; meanwhile, the pure petroleum ether recovered by the rotary evaporator is used for extracting the earthworm active crude fat.
The method is advanced and scientific, and the technical problems to be solved by the invention are as follows: (1) green enzymolysis of earthworm body. Fully mobilize the activity of the earthworm autolytic enzyme and reduce the usage amount of chemicals in the earthworm enzymolysis process. (2) High-efficiency extraction of earthworm body fat substances. The earthworm bodies fully release fat, and simultaneously fat substances are effectively extracted and collected. (3) The biological activity of the earthworm body fat substances is efficiently maintained. The biological activity of the earthworm body fat substances is kept while the earthworm body fat substances are lifted, and a biological function support is provided for subsequent high-added-value utilization.
The invention has the beneficial effects that:
(1) according to the method, the yield of the earthworm crude fat (F1) is 3.6-4.2% of the fresh weight of the earthworms. Meanwhile, the earthworms are inactivated by high temperature (105 ℃) for 30 minutes, dried by hot dry air (75 ℃), extracted by a Soxhlet extraction method and the same extraction technical parameters, and the yield of the crude earthworm fat (F2) is 2.2-2.3% of the fresh weight of the earthworms. The extraction amount of F1 crude fat is 1.64-1.83 times of that of F2.
(2) The inhibition effect of crude earthworm fat on escherichia coli was determined by the tube-dish method, using tap water as a Control (CK). The result shows that the CK has no obvious inhibition zone, the average diameter of the F1 inhibition zone is 17.3-18.6 mm, the average diameter of the F2 inhibition zone is 6.8-7.3 mm, and the diameter of the F1 inhibition zone is 2.47-2.55 times of that of the F2.
(3) When the activity of earthworm crude fat superoxide dismutase (SOD) is measured, the SOD enzyme activity of F1 is 2.36-2.65 units/g.Pr, the SOD enzyme activity of F2 is 0.61-0.63 units/g.Pr, and the SOD enzyme activity of F1 is 3.75-4.21 times of that of F2.
The crude earthworm fat obtained by the technical method has the characteristics of high yield and high biological activity, and lays a biological foundation for further processing and utilization of the crude earthworm fat.
The invention adopts the technical method of enzymolysis, low-temperature drying and organic extraction to extract the active fat in the earthworm body and provides raw materials for the subsequent deep processing of earthworm fat products. At present, the development and utilization of earthworms in the market are mainly focused on active enzymes, and the development and utilization of fat are neglected. The patent has high market implementation possibility and remarkable expected economic benefit.
Drawings
FIG. 1 is a schematic flow chart of the present invention.
Detailed description of the invention
The invention is further described with reference to specific examples. The scope of the invention is not limited thereto:
case 1:
(1) clearing intestines by earthworms. Fresh live earthworms are put into tap water, so that the earthworms can spit out materials swallowed into cavities and intestines in the tap water, and the dosage of the tap water is 2 times of the weight of the earthworms. Standing for 6 hours, fishing out the earthworms, cleaning by flowing tap water, and removing impurities for later use.
(2) Earthworm autolysis enzymolysis. And (3) putting the earthworms after clearing the intestines into a heat-preserving barrel, and uniformly adding a sugar preparation (sucrose: glucose: fructose = 5: 3: 4, weight ratio) in an amount of 4% of the weight of the fresh earthworms. Forcibly stirring the mixture of earthworm body and sugar preparation at 5 r/min while maintaining the barrel temperature of 55 deg.C for 24 hr, filtering with 5 mm screen to remove impurities, and collecting the earthworm enzymolysis liquid.
(3) And (5) freeze-drying the earthworms. And (3) putting the earthworm enzymolysis liquid into a vacuum freeze drying box, and freeze drying to obtain freeze-dried earthworm powder, wherein the water content of the freeze-dried powder is required to be lower than 2%.
(4) Extracting earthworm fat. Petroleum ether with a low boiling range (30-60 ℃) is used as an extracting agent. Filling the freeze-dried powder into a filter paper bag, and putting the filter paper bag into an extraction tube; pouring 1/2 volumes of extractant into a fat burning bottle of a Soxhlet extractor, connecting each part of the Soxhlet extractor, heating at constant temperature (55 ℃) and performing reflux for 4 hours to obtain a mixture of earthworm crude fat and petroleum ether; transferring the mixture to a rotary evaporator, and recovering pure petroleum ether at constant temperature (55 ℃) to obtain a concentrated earthworm crude fat and petroleum ether mixture; and (3) putting the mixture in a fume hood, and exhausting and volatilizing for 12 hours to fully volatilize the residual petroleum ether to obtain the crude earthworm fat. Meanwhile, the pure petroleum ether recovered by the rotary evaporator is used for extracting the earthworm active crude fat.
Has the advantages that:
(1) according to the above method, the yield of crude earthworm fat (F1) was 3.6% of the fresh weight of earthworms. Meanwhile, the earthworms are inactivated by high temperature (105 ℃) for 30 minutes, dried by hot dry air (75 ℃), extracted by a Soxhlet extraction method and the same extraction technical parameters, and the yield of the crude earthworm fat (F2) is 2.2 percent of the fresh weight of the earthworms. The extraction amount of F1 crude fat was 1.64 times that of F2.
(2) The inhibition effect of crude earthworm fat on escherichia coli was determined by the tube-dish method, using tap water as a Control (CK). The result shows that the CK has no obvious inhibition zone, the average diameter of the inhibition zone of F1 is 18.6mm, the average diameter of the inhibition zone of F2 is 7.3mm, and the diameter of the inhibition zone of F1 is 2.55 times of that of F2.
(3) The superoxide dismutase (SOD) activities of F1 and F2 are respectively 2.65units/g.Pr and 0.63units/g.Pr, and the SOD enzyme activity of F1 is 4.21 times that of F2.
Case 2:
(1) clearing intestines by earthworms. Fresh live earthworms are put into tap water, so that the earthworms can spit out materials swallowed into cavities and intestines in the tap water, and the dosage of the tap water is 1 time of the weight of the earthworms. Standing for 8 hours, fishing out the earthworms, cleaning by flowing tap water, and removing impurities for later use.
(2) Earthworm autolysis enzymolysis. And (3) putting the earthworms after clearing the intestines into a heat-preserving barrel, and uniformly adding a sugar preparation (sucrose: glucose: fructose = 3: 1: 2, weight ratio) in an amount of 2% of the weight of the fresh earthworms. Forcibly stirring the mixture of earthworm body and sugar preparation at 5 r/min while keeping the barrel temperature at 45 deg.C for 12 hr, filtering with 5 mm screen to remove impurities, and collecting the earthworm enzymolysis solution.
(3) And (5) freeze-drying the earthworms. And (3) putting the earthworm enzymolysis liquid into a vacuum freeze drying box, and freeze drying to obtain freeze-dried earthworm powder, wherein the water content of the freeze-dried powder is required to be lower than 2%.
(4) Extracting earthworm fat. And extracting fat in the earthworm freeze-dried powder by a Soxhlet extraction method. Petroleum ether with a low boiling range (30-60 ℃) is used as an extracting agent. Filling the freeze-dried powder into a filter paper bag, and putting the filter paper bag into an extraction tube; pouring 1/2 volumes of extractant into a fat bottle of a Soxhlet extractor, connecting each part of the Soxhlet extractor, heating at constant temperature (45 ℃) and distilling for 4 hours to obtain a mixture of earthworm crude fat and petroleum ether; transferring the mixture to a rotary evaporator, and recovering pure petroleum ether at constant temperature (45 ℃) to obtain a concentrated earthworm crude fat and petroleum ether mixture; and (3) putting the mixture in a fume hood, and exhausting and volatilizing for 12 hours to fully volatilize the residual petroleum ether to obtain the crude earthworm fat. Meanwhile, the pure petroleum ether recovered by the rotary evaporator is used for extracting the earthworm active crude fat.
Has the advantages that:
(1) according to the above method, the yield of crude earthworm fat (F1) was 4.2% of the fresh weight of earthworms. Meanwhile, the earthworms are inactivated by high temperature (105 ℃) for 30 minutes, dried by hot dry air (75 ℃), extracted by a Soxhlet extraction method and the same extraction technical parameters, and the yield of the crude earthworm fat (F2) is 2.3 percent of the fresh weight of the earthworms. The extraction amount of F1 crude fat was 1.83 times of that of F2.
(2) The inhibition effect of crude earthworm fat on escherichia coli was determined by the tube-dish method, using tap water as a Control (CK). The result shows that CK has no obvious inhibition zone, the average diameter of the inhibition zone of F1 is 17.3mm, the average diameter of the inhibition zone of F2 is 6.8mm, and the diameter of the inhibition zone of F1 is 2.54 times of that of F2.
(3) The superoxide dismutase (SOD) activities of F1 and F2 are respectively 2.43 units/g.Pr and 0.61 units/g.Pr, and the SOD enzyme activity of F1 is 3.98 times that of F2.
Case 3:
(1) clearing intestines by earthworms. Fresh live earthworms are put into tap water, so that the earthworms can spit out materials swallowed into cavities and intestines in the tap water, and the dosage of the tap water is 2 times of the weight of the earthworms. Standing for 7 hours, fishing out the earthworms, cleaning by flowing tap water, and removing impurities for later use.
(2) Earthworm autolysis enzymolysis. And (3) putting the earthworms after clearing the intestines into a heat-preserving barrel, and uniformly adding a sugar preparation (sucrose: glucose: fructose = 4: 2: 3, weight ratio) in an amount of 3% of the weight of the fresh earthworms. Forcibly stirring the mixture of earthworm body and sugar preparation at 5 r/min while keeping the barrel temperature at 50 deg.C for 18 hr, filtering with 5 mm screen to remove impurities, and collecting the earthworm enzymolysis liquid.
(3) And (5) freeze-drying the earthworms. And (3) putting the earthworm enzymolysis liquid into a vacuum freeze drying box, and freeze drying to obtain freeze-dried earthworm powder, wherein the water content of the freeze-dried powder is required to be lower than 2%.
(4) Extracting earthworm fat. And extracting fat in the earthworm freeze-dried powder by a Soxhlet extraction method. Petroleum ether with a low boiling range (30-60 ℃) is used as an extracting agent. Filling the freeze-dried powder into a filter paper bag, and putting the filter paper bag into an extraction tube; pouring 1/2 volumes of extractant into a Soxhlet extractor fat bottle, connecting each part of the Soxhlet extractor, heating at constant temperature (50 ℃) and performing reflux for 4 hours to obtain a mixture of earthworm crude fat and petroleum ether; transferring the mixture to a rotary evaporator, and recovering pure petroleum ether at constant temperature (50 ℃) to obtain a concentrated earthworm crude fat and petroleum ether mixture; and (3) putting the mixture in a fume hood, and exhausting and volatilizing for 12 hours to fully volatilize the residual petroleum ether to obtain the crude earthworm fat. Meanwhile, the pure petroleum ether recovered by the rotary evaporator is used for extracting the earthworm active crude fat.
Has the advantages that:
(1) according to the above method, the yield of crude earthworm fat (F1) was 3.92% of the fresh weight of earthworms. Meanwhile, the earthworms are inactivated by high temperature (105 ℃) for 30 minutes, dried by hot dry air (75 ℃), extracted by a Soxhlet extraction method and the same extraction technical parameters, and the yield of the crude earthworm fat (F2) is 2.2 percent of the fresh weight of the earthworms. The extraction amount of F1 crude fat was 1.78 times of that of F2.
(2) The inhibition effect of crude earthworm fat on escherichia coli was determined by the tube-dish method, using tap water as a Control (CK). The result shows that CK has no obvious inhibition zone, the average diameter of the inhibition zone of F1 is 17.8mm, the average diameter of the inhibition zone of F2 is 7.2 mm, and the diameter of the inhibition zone of F1 is 2.47 times of that of F2.
(3) The superoxide dismutase (SOD) activities of F1 and F2 are respectively 2.36 units/g.Pr and 0.63units/g.Pr, and the SOD enzyme activity of F1 is 3.75 times that of F2.
Claims (4)
1. A method for extracting earthworm active crude fat is characterized by comprising the following steps:
(1) clearing intestines by earthworms; putting fresh earthworms into tap water, and enabling the earthworms to spit out materials swallowed into cavities and intestines in the tap water, wherein the amount of the tap water is 1-2 times of the weight of the earthworms; standing for 6-8 hours, fishing out the earthworms, cleaning with flowing tap water, and removing impurities to obtain the earthworms with cleared intestines for later use;
(2) carrying out autolysis enzymolysis on earthworms; putting the earthworms with the cleared intestines into a heat-preserving barrel, and uniformly adding a sugar preparation, wherein the adding amount of the sugar preparation is 2-4% of the weight of the earthworms with the cleared intestines; forcibly stirring the mixture of the earthworms and the sugar preparation at the stirring speed of 5 revolutions per minute, keeping the temperature of a heat-preserving container at 45-55 ℃, filtering the mixture by using a 5 mm screen to remove impurities after the earthworms are subjected to enzymolysis for 12-24 hours, and collecting an earthworm enzymolysis solution for later use;
(3) freeze-drying earthworms; putting the earthworm enzymolysis liquid into a vacuum freeze drying box, and freeze drying to obtain earthworm freeze-dried powder;
(4) fat extraction; and extracting fat in the earthworm freeze-dried powder by a Soxhlet extraction method.
2. The method for extracting crude earthworm fat according to claim 1, wherein in the step (2), the sugar preparation comprises the following components: glucose: fructose, sucrose: glucose: the weight ratio of fructose is 3-5: 1-3: 2 to 4.
3. The method for extracting crude earthworm fat according to claim 1, wherein in the step (3), the water content of the freeze-dried powder is less than 2%.
4. The method for extracting the earthworm active crude fat as claimed in claim 1, wherein the fat in the earthworm freeze-dried powder is extracted by a Soxhlet extraction method in the step (3), and the method comprises the following steps:
step 1), preparing petroleum ether with a low boiling range of 30-60 ℃ as an extracting agent; putting the freeze-dried earthworm powder into a filter paper bag, sealing, and putting into an extraction tube; pouring 1/2 volumes of the extracting agent into a fat bottle of a Soxhlet extractor, and connecting the Soxhlet extractor;
step 2), heating the fat bottle at a constant temperature of 45-55 ℃, evaporating and gasifying an extractant in the fat bottle, feeding the extractant into a condensing tube, condensing the extractant, dripping the condensed extractant into the extracting tube to extract fat in the earthworm freeze-dried powder, and refluxing the extractant into the fat bottle when the upper liquid level of the extractant in the extracting tube is higher than a reflux tube;
step 3), carrying out reciprocating reflux extraction for 4 hours according to the step 2) to obtain a mixture of crude earthworm fat and petroleum ether;
step 4), transferring the mixture to a rotary evaporator, and recovering pure petroleum ether at constant temperature of 45-55 ℃ to obtain a concentrated earthworm crude fat and petroleum ether mixture;
step 5), placing the concentrated mixture of the crude earthworm fat and the petroleum ether in a fume hood, and exhausting and volatilizing for 12 hours to fully volatilize the residual petroleum ether to obtain the crude earthworm fat; meanwhile, the pure petroleum ether recovered by the rotary evaporator is used for extracting the earthworm active crude fat.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110365302.9A CN113061482A (en) | 2021-04-06 | 2021-04-06 | A kind of method for extracting earthworm active crude fat |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN202110365302.9A CN113061482A (en) | 2021-04-06 | 2021-04-06 | A kind of method for extracting earthworm active crude fat |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1991007480A1 (en) * | 1989-11-13 | 1991-05-30 | Mta Szegedi Biológiai Központ | Method for the preparation of fat mixtures comprising eicosapentaenoate ester from worms of family lumbricidae and, optionally fatty acid mixtures comprising eicosapentaenoic acid therefrom |
| CN104593456A (en) * | 2014-12-02 | 2015-05-06 | 高向阳 | Method for preparing peptone from enzymatically-hydrolyzed earthworm dry powder and purpose of peptone |
| CN110540550A (en) * | 2019-07-31 | 2019-12-06 | 倪成 | combined preparation process of lumbrokinase and crude earthworm lecithin |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1991007480A1 (en) * | 1989-11-13 | 1991-05-30 | Mta Szegedi Biológiai Központ | Method for the preparation of fat mixtures comprising eicosapentaenoate ester from worms of family lumbricidae and, optionally fatty acid mixtures comprising eicosapentaenoic acid therefrom |
| CN104593456A (en) * | 2014-12-02 | 2015-05-06 | 高向阳 | Method for preparing peptone from enzymatically-hydrolyzed earthworm dry powder and purpose of peptone |
| CN110540550A (en) * | 2019-07-31 | 2019-12-06 | 倪成 | combined preparation process of lumbrokinase and crude earthworm lecithin |
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