CN105266075B - method for extracting Stichopus japonicus saponins from Stichopus japonicus blanching liquid - Google Patents
method for extracting Stichopus japonicus saponins from Stichopus japonicus blanching liquid Download PDFInfo
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- CN105266075B CN105266075B CN201510693837.3A CN201510693837A CN105266075B CN 105266075 B CN105266075 B CN 105266075B CN 201510693837 A CN201510693837 A CN 201510693837A CN 105266075 B CN105266075 B CN 105266075B
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- 239000007788 liquid Substances 0.000 title claims abstract description 45
- 206010033546 Pallor Diseases 0.000 title claims abstract description 40
- 229930182490 saponin Natural products 0.000 title claims abstract description 37
- 150000007949 saponins Chemical class 0.000 title claims abstract description 37
- 238000000034 method Methods 0.000 title claims abstract description 34
- 235000017709 saponins Nutrition 0.000 title claims description 35
- 241000965254 Apostichopus japonicus Species 0.000 title claims description 5
- 241000251511 Holothuroidea Species 0.000 claims abstract description 79
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 75
- 239000007787 solid Substances 0.000 claims abstract description 33
- 239000000843 powder Substances 0.000 claims abstract description 29
- 239000001397 quillaja saponaria molina bark Substances 0.000 claims abstract description 28
- 238000000605 extraction Methods 0.000 claims abstract description 23
- 238000001179 sorption measurement Methods 0.000 claims abstract description 22
- 239000011347 resin Substances 0.000 claims abstract description 21
- 229920005989 resin Polymers 0.000 claims abstract description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 20
- 239000012530 fluid Substances 0.000 claims abstract description 13
- 239000006228 supernatant Substances 0.000 claims abstract description 12
- 239000003480 eluent Substances 0.000 claims abstract description 11
- 238000000967 suction filtration Methods 0.000 claims abstract description 11
- 239000000284 extract Substances 0.000 claims abstract description 9
- 238000009777 vacuum freeze-drying Methods 0.000 claims abstract description 6
- 239000012065 filter cake Substances 0.000 claims description 15
- 239000000706 filtrate Substances 0.000 claims description 11
- 238000009835 boiling Methods 0.000 claims description 10
- 239000002994 raw material Substances 0.000 claims description 10
- 238000001694 spray drying Methods 0.000 claims description 10
- 238000002156 mixing Methods 0.000 claims description 7
- 239000004519 grease Substances 0.000 claims description 6
- 238000001042 affinity chromatography Methods 0.000 claims description 5
- 238000005238 degreasing Methods 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 5
- 239000000469 ethanolic extract Substances 0.000 claims description 5
- 210000000936 intestine Anatomy 0.000 claims description 5
- 239000002244 precipitate Substances 0.000 claims description 5
- 239000008213 purified water Substances 0.000 claims description 5
- 238000004062 sedimentation Methods 0.000 claims description 5
- 230000002745 absorbent Effects 0.000 claims 1
- 239000002250 absorbent Substances 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 abstract description 7
- 229930183167 cerebroside Natural products 0.000 abstract description 5
- 150000003904 phospholipids Chemical class 0.000 abstract description 5
- 239000000047 product Substances 0.000 abstract description 4
- JSPNNZKWADNWHI-PNANGNLXSA-N (2r)-2-hydroxy-n-[(2s,3r,4e,8e)-3-hydroxy-9-methyl-1-[(2r,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoctadeca-4,8-dien-2-yl]heptadecanamide Chemical compound CCCCCCCCCCCCCCC[C@@H](O)C(=O)N[C@H]([C@H](O)\C=C\CC\C=C(/C)CCCCCCCCC)CO[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O JSPNNZKWADNWHI-PNANGNLXSA-N 0.000 abstract description 2
- RIZIAUKTHDLMQX-UHFFFAOYSA-N cerebroside D Natural products CCCCCCCCCCCCCCCCC(O)C(=O)NC(C(O)C=CCCC=C(C)CCCCCCCCC)COC1OC(CO)C(O)C(O)C1O RIZIAUKTHDLMQX-UHFFFAOYSA-N 0.000 abstract description 2
- 238000004440 column chromatography Methods 0.000 abstract description 2
- 150000001875 compounds Chemical class 0.000 abstract description 2
- 239000012141 concentrate Substances 0.000 abstract description 2
- 238000010924 continuous production Methods 0.000 abstract description 2
- 238000005377 adsorption chromatography Methods 0.000 abstract 1
- 239000013543 active substance Substances 0.000 description 3
- 150000001784 cerebrosides Chemical class 0.000 description 3
- 230000000694 effects Effects 0.000 description 2
- 238000003912 environmental pollution Methods 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- 208000005189 Embolism Diseases 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000700721 Hepatitis B virus Species 0.000 description 1
- 241000725303 Human immunodeficiency virus Species 0.000 description 1
- 206010029113 Neovascularisation Diseases 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 241000700584 Simplexvirus Species 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- 230000000843 anti-fungal effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000003471 anti-radiation Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 206010003246 arthritis Diseases 0.000 description 1
- 230000002358 autolytic effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 230000020764 fibrinolysis Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 229940024999 proteolytic enzymes for treatment of wounds and ulcers Drugs 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/616—Echinodermata, e.g. starfish, sea cucumbers or sea urchins
Landscapes
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Medicines Containing Plant Substances (AREA)
- Extraction Or Liquid Replacement (AREA)
- Preparation Of Fruits And Vegetables (AREA)
- Compounds Of Unknown Constitution (AREA)
Abstract
The invention discloses a method for extracting holothurian saponin from holothurian blanching liquids, which is to decompress and concentrate the holothurian blanching liquids, spray-dry the holothurian blanching liquids and then use supercritical CO2Extracting the fluid to obtain solid powder with oil removed, performing alcohol extraction, suction filtration and concentration on the solid powder, re-dissolving the solid powder in water, removing compounds such as phospholipid, cerebroside and the like in the solution by adopting a centrifugal method, performing adsorption chromatography on the obtained supernatant through a macroporous adsorption resin column, eluting by using an ethanol solution, and performing vacuum freeze drying on the eluent to obtain the sea cucumber saponin extract. The invention improves the adsorption efficiency of macroporous adsorption resin column chromatography, prolongs the service life of the resin column, has reasonable process, simple operation and low production cost, can realize continuous production, and improves the purity and the yield of target products.
Description
Technical Field
The invention relates to a technology for extracting active substances from sea cucumber processing byproducts, in particular to a method for extracting sea cucumber saponin from sea cucumber blanching liquid.
Background
The fresh sea cucumber is easy to peel and dissolve due to the existence of proteolytic enzymes such as autolytic enzyme and the like in the fresh sea cucumber after being caught, and the fresh sea cucumber is processed and stored after being blanched by hot water for convenient storage, however, the fresh sea cucumber generates a large amount of blanching liquid in the hot water treatment process, the blanching liquid of the sea cucumber contains a large amount of active substances such as sea cucumber saponin through detection, the sea cucumber saponin is main active ingredients in the sea cucumber and has various pharmacological activities such as anti-tumor effect, immunity improvement, antifungal effect, neovascularization resistance, fibrinolysis promotion, embolism formation inhibition, anti-inflammatory effect (such as arthritis), anti-radiation activity, AIDS virus, hepatitis B virus, herpes simplex virus and the like, and the recent research finds that the sea cucumber saponin also participates in fat metabolism and has the effect of losing weight.
For example, CN1733793A published in 2.15.2006 discloses a process for preparing holothurian saponins by using sea cucumber processing waste liquid, which comprises the steps of filtering the sea cucumber waste liquid, precipitating the filtrate with ethanol, standing, centrifuging, concentrating the supernatant under reduced pressure to remove ethanol components to obtain a concentrated solution, passing the concentrated solution through a macroporous adsorption resin column at a flow rate of 1.5-2.0 BV/h, eluting with water at the same flow rate to remove impurity components such as salts, saccharides, pigments and the like, eluting the holothurian saponins adsorbed on the macroporous adsorption resin with 60-70% ethanol, collecting the eluate, concentrating the eluate under reduced pressure to obtain an extract, and freeze-drying to obtain a holothurian saponin dry product.
Disclosure of Invention
In order to overcome the defects of low adsorption efficiency, weak adsorption specificity, easy column blockage and low saponin content purity existing in the process of directly adsorbing the saponin in the sea cucumber blanching liquid by using macroporous adsorption resin in the prior art, the invention provides methods for extracting the sea cucumber saponin by using the sea cucumber blanching liquid, which have the advantages of reasonable process, feasible operation, low production cost, high product yield and high purity.
The technical scheme adopted by the invention for solving the technical problems is that methods for extracting holothurian saponins by utilizing a holothurian blanching liquid are characterized by comprising the following process steps:
(1) selection and treatment of raw materials
Selecting a sea cucumber blanching liquid as a raw material, concentrating the sea cucumber blanching liquid under reduced pressure until the solid content is 20-45% to obtain a concentrated liquid;
(2) spray drying
Spray-drying the concentrated solution obtained in the step (1) to obtain th solid powder;
(3) degreasing
Subjecting the th solid powder obtained in step (2) to supercritical CO2Extracting the fluid, and removing grease to obtain second solid powder;
(4) alcohol extraction
Adding the second solid powder obtained in the step (3) into an ethanol solution with the mass being 20-100 times that of the second solid powder, and extracting for 12-24 hours to obtain an extracting solution; wherein the concentration of the ethanol solution is 70-95%;
(5) suction filtration and concentration
Carrying out suction filtration on the extracting solution obtained in the step (4) to obtain a filtrate; extracting the filter cake for 12-24 h by using 70-95% ethanol solution again to obtain filter cake ethanol extract; repeating the step for 2 times; mixing all the filtrate and the filter cake alcohol extract, concentrating under reduced pressure, and removing ethanol to obtain concentrated solution;
(6) water sedimentation
Adding purified water with the volume of 100-300 times of that of the concentrated solution obtained in the step (5) into the concentrated solution for dissolving, and removing precipitates through a horizontal screw centrifuge, a disc centrifuge and a tubular centrifuge in sequence to obtain supernatant for later use;
(7) affinity chromatography
Adsorbing and chromatographing the supernatant obtained in the step (6) through a macroporous adsorption resin column, and eluting the macroporous adsorption resin column by using an ethanol solution with the concentration of 60-70% to obtain an eluent;
(8) concentrating and drying
And (4) carrying out vacuum freeze drying on the eluent obtained in the step (7) to obtain the sea cucumber saponin extract.
The sea cucumber blanching liquid is obtained by dissecting and removing intestines of fresh sea cucumbers, putting the fresh sea cucumbers into boiling water, boiling for 10-30 min, and fishing out the sea cucumbers.
The supercritical CO2The temperature of the fluid extraction is controlled to be 40-50 ℃, the pressure is 30-40 Mpa, and the extraction time is 1-2 h.
The macroporous adsorbent resin column is AB-8 type or D101 type.
The invention is to concentrate the sea cucumber blanching liquid, spray-dry and then utilize supercritical CO2The fluid extraction, the fluid of the supercritical state has super strong dissolving capacity, the grease substances in the sea cucumber blanching liquid are effectively removed, the oil mixing in the subsequent extraction process is avoided, the degreased sea cucumber blanching liquid powder is extracted by an alcohol precipitation water extraction method, firstly, an ethanol solution is utilized for extraction, an extracting solution is re-dissolved in water after being concentrated, a centrifugal method is utilized for removing compounds such as phospholipid, cerebroside and the like in the solution, the mixing of impurities in the subsequent extraction process is further reduced by steps, the obtained extracting solution is high in clarity, the macroporous adsorption resin column chromatography adsorption efficiency is improved, the service life of a resin column is prolonged, the continuous production can be realized, the purity of a target product is improved, the sea cucumber saponin extracted by the method is high, the total saponin content is between 90 and 95 percent, and the technical method increases the operability and practicability of extracting the saponin from the sea cucumber blanching liquid, and solves the problems that the macroporous resin is directly utilized for adsorbing the saponin in the blanching liquid, the adsorption is low in adsorption efficiency,The extraction method has the advantages of reasonable process, simple operation and low production cost, and the extracted active substance saponin has higher health care value, reduces the environmental pollution after the discharge of the sea blanching liquid, and has universal social and economic benefits.
Detailed Description
The invention is further illustrated with reference to the following examples.
Example 1
method for extracting holothurian saponin by utilizing holothurian blanching liquid, which comprises the following process steps:
(1) selection and treatment of raw materials
Selecting a sea cucumber blanching liquid as a raw material, carrying out reduced pressure concentration on the sea cucumber blanching liquid under the pressure of 100 atm, and concentrating until the solid content is 30% to obtain a concentrated solution; wherein the sea cucumber blanching liquid is prepared by dissecting fresh sea cucumber, removing intestine, boiling in boiling water for 20min, and taking out sea cucumber;
(2) spray drying
Spray-drying the concentrated solution obtained in the step (1) to obtain th solid powder;
(3) degreasing
Subjecting the th solid powder obtained in step (2) to supercritical CO2Extracting the fluid, and removing grease to obtain second solid powder; wherein, supercritical CO2The temperature of fluid extraction is controlled to be 45 ℃, the pressure is 35Mpa, and the extraction time is 1.5 h;
(4) alcohol extraction
Adding the second solid powder obtained in the step (3) into an ethanol solution with the mass being 80 times that of the second solid powder, and extracting for 15 hours to obtain an extracting solution; wherein the concentration of the ethanol solution is 90 percent;
(5) suction filtration and concentration
Carrying out suction filtration on the extracting solution obtained in the step (4) to obtain a filtrate; extracting the filter cake for 15h by using 90% ethanol solution again to obtain filter cake ethanol extract; repeating the step for 2 times; mixing all the filtrates and the filter cake ethanol extractive solution, concentrating under reduced pressure below 30 deg.C, and removing ethanol to obtain concentrated solution;
(6) water sedimentation
Dissolving the concentrated solution obtained in the step (5) in purified water 200 times the volume of the concentrated solution, sequentially removing phospholipids and cerebrosides precipitates through a horizontal screw centrifuge (rotating speed 3000 rpm), a disc centrifuge (rotating speed 6000 rpm) and a tubular centrifuge (rotating speed 12000 rpm), and obtaining a supernatant for later use;
(7) affinity chromatography
Adsorbing and chromatographing the supernatant obtained in the step (6) by an AB-8 type macroporous adsorption resin column, and eluting the macroporous adsorption resin column by using 65% ethanol solution to obtain eluent;
(8) concentrating and drying
And (4) carrying out vacuum freeze drying on the eluent obtained in the step (7) to obtain the sea cucumber saponin extract.
The extraction method provided by the embodiment has the advantages of reasonable process, simple operation and low production cost, the holothurian saponin extracted by the method has high purity, and the total saponin content is 92 percent through determination, meanwhile, the environmental pollution after the discharge of the sea blanching liquid is reduced, and the social and economic benefits of are achieved.
Example 2
method for extracting holothurian saponin by utilizing holothurian blanching liquid, which comprises the following process steps:
(1) selection and treatment of raw materials
Selecting a sea cucumber blanching liquid as a raw material, carrying out reduced pressure concentration on the sea cucumber blanching liquid under the pressure of 70 atmospheric pressure, and concentrating until the solid content is 45% to obtain a concentrated solution; wherein the sea cucumber blanching liquid is prepared by dissecting fresh sea cucumber, removing intestine, boiling in boiling water for 30min, and taking out sea cucumber;
(2) spray drying
Spray-drying the concentrated solution obtained in the step (1) to obtain th solid powder;
(3) degreasing
Subjecting the th solid powder obtained in step (2) to supercritical CO2Extracting the fluid, and removing grease to obtain second solid powder; wherein, supercritical CO2Temperature of fluid extractionControlling the temperature at 50 deg.C, the pressure at 40Mpa, and the extraction time at 2 h;
(4) alcohol extraction
Adding the second solid powder obtained in the step (3) into an ethanol solution with the mass being 100 times that of the second solid powder, and extracting for 24 hours to obtain an extracting solution; wherein the concentration of the ethanol solution is 70 percent;
(5) suction filtration and concentration
Carrying out suction filtration on the extracting solution obtained in the step (4) to obtain a filtrate; extracting the filter cake for 24h by using 70% ethanol solution again to obtain filter cake ethanol extract; repeating the step for 2 times; mixing all the filtrates and the filter cake ethanol extractive solution, concentrating under reduced pressure below 25 deg.C, and removing ethanol to obtain concentrated solution;
(6) water sedimentation
Dissolving the concentrated solution obtained in the step (5) by adding purified water 300 times of the volume of the concentrated solution, and removing phospholipids and cerebrosides precipitates by using a horizontal screw centrifuge (rotating speed 3000 rpm), a disc centrifuge (rotating speed 6000 rpm) and a tubular centrifuge (rotating speed 12000 rpm) to obtain a supernatant for later use;
(7) affinity chromatography
Adsorbing and chromatographing the supernatant obtained in the step (6) by using a D101 type macroporous adsorption resin column, and eluting the macroporous adsorption resin column by using 70% ethanol solution to obtain eluent;
(8) concentrating and drying
And (4) carrying out vacuum freeze drying on the eluent obtained in the step (7) to obtain the sea cucumber saponin extract.
The extraction method provided by the embodiment has the advantages of reasonable process, simple operation and low production cost. The holothurian saponin extracted by the method has high purity, and is determined as follows: the content of total saponins is 95%.
Example 3
method for extracting holothurian saponin by utilizing holothurian blanching liquid, which comprises the following process steps:
(1) selection and treatment of raw materials
Selecting a sea cucumber blanching liquid as a raw material, carrying out reduced pressure concentration on the sea cucumber blanching liquid under the pressure of 200 atm, and concentrating until the solid content is 20% to obtain a concentrated solution; wherein the sea cucumber blanching liquid is obtained by dissecting fresh sea cucumber, removing intestines, boiling in boiling water for 10min, and taking out the sea cucumber;
(2) spray drying
Spray-drying the concentrated solution obtained in the step (1) to obtain th solid powder;
(3) degreasing
Subjecting the th solid powder obtained in step (2) to supercritical CO2Extracting the fluid, and removing grease to obtain second solid powder; wherein, supercritical CO2The temperature of fluid extraction is controlled at 40 ℃, the pressure is 30Mpa, and the extraction time is 1 h;
(4) alcohol extraction
Adding the second solid powder obtained in the step (3) into an ethanol solution with the mass 20 times of that of the second solid powder, and extracting for 12 hours to obtain an extracting solution; wherein the concentration of the ethanol solution is 95 percent;
(5) suction filtration and concentration
Carrying out suction filtration on the extracting solution obtained in the step (4) to obtain a filtrate; extracting the filter cake for 12h by using 95% ethanol solution again to obtain filter cake ethanol extract; repeating the step for 2 times; mixing all the filtrates and the filter cake ethanol extractive solution, concentrating under reduced pressure below 30 deg.C, and removing ethanol to obtain concentrated solution;
(6) water sedimentation
Dissolving the concentrated solution obtained in the step (5) in purified water with the volume 100 times of that of the concentrated solution, and removing phospholipids and cerebrosides precipitates by a horizontal screw centrifuge (rotating speed 3000 rpm), a disc centrifuge (rotating speed 6000 rpm) and a tubular centrifuge (rotating speed 12000 rpm) to obtain a supernatant for later use;
(7) affinity chromatography
Adsorbing and chromatographing the supernatant obtained in the step (6) by an AB-8 type macroporous adsorption resin column, and eluting the macroporous adsorption resin column by using 60% ethanol solution to obtain eluent;
(8) concentrating and drying
And (4) carrying out vacuum freeze drying on the eluent obtained in the step (7) to obtain the sea cucumber saponin extract.
The extraction method provided by the embodiment has the advantages of reasonable process, simple operation and low production cost. The holothurian saponin extracted by the method has high purity, and is determined as follows: the content of total saponins is 90%.
Claims (4)
1. method for extracting holothurian saponin by utilizing holothurian blanching liquid, which is characterized by comprising the following process steps:
(1) selection and treatment of raw materials
Selecting a sea cucumber blanching liquid as a raw material, concentrating the sea cucumber blanching liquid under reduced pressure until the solid content is 20-45% to obtain a concentrated liquid;
(2) spray drying
Spray-drying the concentrated solution obtained in the step (1) to obtain th solid powder;
(3) degreasing
Subjecting the th solid powder obtained in step (2) to supercritical CO2Extracting the fluid, and removing grease to obtain second solid powder;
(4) alcohol extraction
Adding the second solid powder obtained in the step (3) into an ethanol solution with the mass being 20-100 times that of the second solid powder, and extracting for 12-24 hours to obtain an extracting solution; wherein the concentration of the ethanol solution is 70-95%;
(5) suction filtration and concentration
Carrying out suction filtration on the extracting solution obtained in the step (4) to obtain a filtrate; extracting the filter cake for 12-24 h by using 70-95% ethanol solution again to obtain filter cake ethanol extract; repeating the step for 2 times; mixing all the filtrate and the filter cake alcohol extract, concentrating under reduced pressure, and removing ethanol to obtain concentrated solution;
(6) water sedimentation
Adding purified water with the volume of 100-300 times of that of the concentrated solution obtained in the step (5) into the concentrated solution for dissolving, and removing precipitates through a horizontal screw centrifuge, a disc centrifuge and a tubular centrifuge in sequence to obtain supernatant for later use;
(7) affinity chromatography
Adsorbing and chromatographing the supernatant obtained in the step (6) through a macroporous adsorption resin column, and eluting the macroporous adsorption resin column by using an ethanol solution with the concentration of 60-70% to obtain an eluent;
(8) concentrating and drying
And (4) carrying out vacuum freeze drying on the eluent obtained in the step (7) to obtain the sea cucumber saponin extract.
2. The method for extracting the holothurian saponins from the holothurian blanching liquids according to claim 1, wherein the holothurian blanching liquids are liquids obtained by dissecting fresh holothurian, removing intestines, putting the fresh holothurian into boiling water for boiling for 10-30 min, and fishing out the holothurian.
3. The method for extracting Stichopus japonicus selenka blanching liquid according to claim 1, wherein the supercritical CO is used2The temperature of the fluid extraction is controlled to be 40-50 ℃, the pressure is 30-40 Mpa, and the extraction time is 1-2 h.
4. The method for extracting the holothurian saponins from holothurian blanching liquid according to claim 1, which is characterized in that the macroporous absorbent resin column is AB-8 type or D101 type.
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| CN110256524A (en) * | 2019-07-19 | 2019-09-20 | 荣成市慧海创达生物科技有限公司 | The extracting method of selenka |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104262451A (en) * | 2014-10-24 | 2015-01-07 | 威海芝恩药业股份有限公司 | Method for extracting holothurin from sea cucumber processing waste fluid |
| CN104402983A (en) * | 2014-12-11 | 2015-03-11 | 山东省科学院生物研究所 | Method for extracting glycoprotein by utilizing sea cucumber treatment blanching liquid and application thereof |
| CN104739868A (en) * | 2015-02-15 | 2015-07-01 | 大连工业大学 | Comprehensive utilization method for sea cucumber processing waste liquid |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104262451A (en) * | 2014-10-24 | 2015-01-07 | 威海芝恩药业股份有限公司 | Method for extracting holothurin from sea cucumber processing waste fluid |
| CN104402983A (en) * | 2014-12-11 | 2015-03-11 | 山东省科学院生物研究所 | Method for extracting glycoprotein by utilizing sea cucumber treatment blanching liquid and application thereof |
| CN104739868A (en) * | 2015-02-15 | 2015-07-01 | 大连工业大学 | Comprehensive utilization method for sea cucumber processing waste liquid |
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