WO2007041951A1 - Procede d'extraction du polysaccharide de l'ormeau - Google Patents

Procede d'extraction du polysaccharide de l'ormeau Download PDF

Info

Publication number
WO2007041951A1
WO2007041951A1 PCT/CN2006/002650 CN2006002650W WO2007041951A1 WO 2007041951 A1 WO2007041951 A1 WO 2007041951A1 CN 2006002650 W CN2006002650 W CN 2006002650W WO 2007041951 A1 WO2007041951 A1 WO 2007041951A1
Authority
WO
WIPO (PCT)
Prior art keywords
abalone
minutes
supernatant
water
hours
Prior art date
Application number
PCT/CN2006/002650
Other languages
English (en)
Chinese (zh)
Inventor
Beiwei Zhu
Dongmei Li
Hongling Yin
Xiuping Dong
Yu Song
Yueyue Li
Dan Jiang
Original Assignee
Dalian Polytechnic University
Dalian Zhangzidao Fishery Group Co., Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Dalian Polytechnic University, Dalian Zhangzidao Fishery Group Co., Ltd. filed Critical Dalian Polytechnic University
Priority to JP2008533850A priority Critical patent/JP4420470B2/ja
Publication of WO2007041951A1 publication Critical patent/WO2007041951A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the invention relates to a method for extracting, separating and purifying polysaccharides, belonging to the field of biological products.
  • Background Art In recent years, with the development of molecular biology, it has been gradually recognized that polysaccharides, proteins and nucleic acids are one of the three types of biological macromolecules involved in the nature of life activities. Polysaccharide is a kind of biological macromolecular substance with a wide range of biological activities. It is also called polysaccharide. It is not only an essential component of human life, but also exists in all cell membrane structures and participates in various life function activities. In recent years, it has been found that polysaccharides have enhanced immune function to the human body, as well as physiological activities such as anti-cancer, anti-radiation, anti-inflammatory, and hypoglycemic effects, and thus have received extensive attention and attention.
  • the object of the present invention is to provide a process for efficiently extracting abalone polysaccharides from husked abalone whole, abalone gastropods and abalone organs, and purifying and purifying them.
  • the technical scheme of the invention is as follows: the abalone is obtained by processing, extracting, alcohol precipitation and drying of the raw material to obtain crude abalone polysaccharide; and the crude polysaccharide can be isolated and purified to obtain abalone refined polysaccharide.
  • the specific process steps are - I.
  • Raw material processing Raw abalone can be fresh, frozen, or dried.
  • the abalone meat can be divided into the whole (without the gastropods and organs), the abdomen's gastropods and organs.
  • Abalone polysaccharides can also be extracted separately from the abdomen's gastropods and organs.
  • the method for extracting abalone polysaccharide adopts water leaching method, lye extraction method, ultrasonic extraction method and enzyme extraction method, and the enzyme can be combined with the first three methods to increase the yield.
  • the abalone slurry or the abalone dry powder obtained by the raw material treatment is added with water of 10 to 50 times the amount of the abalone, and ultrasonically treated with 20 to 30 KHZ for 10 to 60 minutes, and the supernatant and the insoluble matter are obtained by centrifugation. Further, 10 to 40 times of water was added to the insoluble matter, and ultrasonic treatment was carried out for 10 to 60 minutes, and the supernatant and the insoluble matter were obtained by centrifugation. The resulting supernatant was combined twice.
  • the enzyme extraction method is abalone slurry or abalone dry powder added with water, and the enzyme is subjected to enzymatic hydrolysis under the conditions of suitable pH and temperature of the enzyme used, and the enzyme used may be single enzyme, double enzyme, complex enzyme or autolytic enzyme, or By combining with the above extraction method, the extraction purpose can be achieved.
  • the single enzyme, the double enzyme, and the complex enzyme are pepsin, trypsin, papain, neutral protease, and the like.
  • the solution is 0. 05 ⁇ 3. 00% of pepsin.
  • the solution is added with a solution of 0. 05 ⁇ 3.
  • Stir at 20 ⁇ 60 °C for 1 ⁇ 6 hours maintain the above pH value during hydrolysis; adjust pH to 7 ⁇ 9 with 0.5mol/L NaOH or K0H solution, add 0.05 ⁇ 3.00% trypsin Stir in the 20 ⁇ 60 °C water bath for 1 ⁇ 6 hours, maintain the above pH value during hydrolysis; adjust the pH to neutral after acid hydrolysis, and heat up to 90 ⁇ 98 °C for 10 minutes in 2 ⁇ 5 minutes.
  • Compound enzyme enzymatic hydrolysis method Add the abalone slurry or abalone dry powder obtained from the raw material treatment, add 10 ⁇ 50 times of water to the abalone, adjust the pH to ⁇ 9 with 0.5mol/L NaOH or K0H solution, and add papain.
  • the amount of addition is 0.05 ⁇ 3.00% of the weight of the solution, the ratio of the three enzymes is 1.0: 0.1-1.0: 0.1-1.0, at a temperature of 20 ⁇ 60 ° C, enzymatic hydrolysis 1 ⁇ 6
  • the pH7 with acid adjust the pH7 with acid, heat up to 90 ⁇ 98 °C for 2 minutes, 1-2 minutes, cool to room temperature for 5 minutes, centrifuge to obtain the supernatant and precipitate; the supernatant is ready for use, the precipitation can also be added 10 ⁇ 40 times of ice is repeated for the above steps 1 ⁇ 2 times, and the supernatant separated by centrifugation is combined for use.
  • Self-dissolving and exogenous enzyme phase-solving (1) Autolysis: It can be autolyzed by any of the following methods: 1 Abalone slurry is added with 10-50 times water weight, and irradiated with ultraviolet light for 10-30 minutes. 0.06 ⁇ 0.08mol/L NaCl is autolyzed by its own enzyme at pH 7.0 7.5, 30 ⁇ 50 °C; 2 The abalone slurry plus abalone 10 ⁇ 50 times water is placed in a sterile enzyme tank at pH6. .0 ⁇ 7.5, self-dissolving at room temperature for 6 ⁇ 8 hours; 3 Put abalone slurry plus 10 ⁇ 50 times water into sterile enzyme tank, at pH 6.0 ⁇ 7.5, 0 ⁇ 4°C autolysis 24 ⁇ 48 hours.
  • the above operation requires two different enzymes for the hydrolysis of the two enzymes. Therefore, if the two enzymes require the same pH value, two enzymes can be added at the same time. For example, when papain, neutral protease and trypsin are hydrolyzed, the pH value is the same, 7 ⁇ 9, and two enzymes can be added at this pH.
  • the centrifugation supernatant obtained above was concentrated in vacuo to a polysaccharide content of 1.5 to 3.0%.
  • the polysaccharide after the alcohol precipitation is centrifuged at a high speed for about 10 minutes, and the obtained precipitate is a crude polysaccharide of abalone.
  • Drying of abalone crude polysaccharide can be achieved by any of the following methods:
  • the purified centrifugal insoluble matter is dried under vacuum at a temperature of 50 to 60 ° C to obtain a dried dried abalone crude polysaccharide.
  • the centrifugally insoluble matter obtained by the purification is diluted with water to a specific gravity of about 1.05 to 1.10, spray-dried, the inlet temperature of the spray drying equipment is 160 to 180 ° C, and the outlet temperature is 50 to 60 ° C to obtain a dried crude abalone polysaccharide.
  • the abalone polysaccharide obtained above may be pulverized to 160 mesh or less by a pulverizer.
  • the abalone polysaccharide extracted by the method of the present invention is a crude polysaccharide, a brown powder, and the polysaccharide extraction rate is 20% to 40% (dry ratio), the polysaccharide content is 6 to 18%, and is easily soluble in hot water, slightly soluble. In cold water. Can be widely used in the food industry. If further purified and deproteinized, the polysaccharide content can reach 40% ⁇ 60%, or even higher, and can be widely used in the pharmaceutical industry.
  • the crude abalone polysaccharide extracted by the above steps can also be further purified and purified according to the following steps:
  • abalone gastropod polysaccharide The crude abalone polysaccharide (hereinafter referred to as abalone gastropod polysaccharide) extracted from the abdomen of abalone is formulated into a 2 ⁇ 10% solution, and a 10% trichloroacetic acid solution is slowly added at 4 ° C, the volume ratio is 5 : 1, Stir. After the shaking at this temperature for 10 minutes, the high-speed centrifugation was carried out for 10 minutes under the conditions of 4 Torr, and the supernatant was subjected to the above-mentioned deproteinization step several times, and the protein content in the polysaccharide solution was determined to be 0.5% or less, and 4 was added to the solution.
  • abalone gastropod polysaccharide The crude abalone polysaccharide extracted from the abdomen of abalone is formulated into a 2 ⁇ 10% solution, and a 10% trichloroacetic acid solution is slowly added at 4 ° C, the volume ratio is 5 : 1, Stir. After the shaking
  • abalone organ polysaccharide The crude abalone polysaccharide (hereinafter referred to as abalone organ polysaccharide) extracted from the abalone organ is formulated into a solution of 2 to 10%, and 0.05 to 1% of pepsin is added to the weight of the solution, and adjusted with 6 mol/L of HC1. After pH 1.5 ⁇ 3.0% and temperature 37 °C, after enzymatic hydrolysis for 1 ⁇ 6 hours, heat up to 90 ⁇ 98 °C for 10 minutes for 2 ⁇ 5 minutes, cool to room temperature, and adjust to neutral with NaOH solution. Centrifuge for 10 minutes, take the supernatant, add 3 ⁇ 4. 5 times the volume of 95% ethanol, at a temperature of 0 ⁇ 4 °C, alcohol precipitation for about 12 ⁇ 16 hours.
  • abalone organ polysaccharide The crude abalone polysaccharide (hereinafter referred to as abalone organ polysaccharide) extracted from the abalone organ is formulated into a solution of 2 to 10%
  • abalone gastropod polysaccharides in addition to glycogen
  • the abalone and abdomen polysaccharides were prepared as a 5% solution, and placed in a 7000Da dialysis bag, and added 0. 05 ⁇ 1% of the ⁇ -amylase at 37 ° C in 0.9% NaCL Dialysis was carried out in the solution, and after hydrolysis for 48 hours, it was dialyzed again in water for 48 hours. After dialysis, the inner liquid of the bag is concentrated, and lyophilized to obtain a white powder, that is, refined abalone gastropod polysaccharide (AGP).
  • AGP refined abalone gastropod polysaccharide
  • AGP was separated by Sephadex G-100 column chromatography.
  • the eluent was 0.9% NaCl solution, the elution rate was 0.5 ml/min, and the phenol sulfuric acid method was used for tracking detection. The only elution peak was drawn and collected. Combine, dialyze, concentrate, freeze-dry to obtain a uniform abalone gastropod polysaccharide AGP.
  • AHP was separated by Sephadex G-100 column chromatography.
  • the eluent was 0.9% NaCl solution, the elution rate was 0.5 ml/min, and the phenol sulfuric acid method was used for tracking detection.
  • Two eluted peaks were drawn and collected separately. The eluted peaks were dialyzed, dried, and then purified by DEAE cellulose 52, and the gradient eluted with a solution of NaCl was a single eluting peak. Collect and collect, dialysis, and concentrate. Freeze-drying yielded uniform abalone organ polysaccharides AHP-1 and AHP-2.
  • the gel column was a Sepharose CL-6B column, and the eluent was 0.9% NaCl solution at a flow rate of 0.24 ml/min.
  • Standard solution preparation Standard glucans with molecular weights of 1000, 5000, 12000, 80,000 and 270,000 respectively were dissolved in double distilled water to prepare a solution of 10 mg/ml, respectively, and the samples were injected according to the molecular weight from small to large, and separated. And collected, phenol sulfuric acid method to track the sugar content, detection at 490nm, record the number of tubes with the largest OD value.
  • Linear regression was performed on the logarithm of the molecular weight by the number of tubes to obtain a linear regression equation.
  • the purified polysaccharide was dissolved in double distilled water to prepare a solution of 10 mg/ml concentration, and the sample was injected under the same conditions, and the number of tubes was recorded, and the molecular weight was calculated by substituting the regression equation. '
  • the chromatographic conditions were: gas chromatograph (Agilent 6890N, USA), HP-1 column, stationary phase Methylsiloxane, carrier gas N2, flow rate 45 ml/min. ⁇ Use hydrogen flame ion detector (FID), detection temperature 300 °C, the injection volume is 1 ⁇ 1.
  • the inlet gasification temperature is 300 °C
  • the column temperature is programmed, 150 °C, lmin, 10 °C/min to 182 °C, 2 min, l °C/min to 188 °C, lmin, 8 °C/min To 230 ° C.
  • the AGP obtained by the method of the present invention is a white powder having a molecular weight of 5000-1000 dal, which is composed of glucose; and the abalone organ polysaccharides AHP-1 and AHP-2 obtained by the method of the present invention are all light brown powder, wherein AHP- The molecular weight of 1 is about 5 X 10 5 dal, which is composed of glucose and arabinose.
  • AHP-1 does not contain amino sugar
  • AHP-2 Traces of N-acetylglucosamine.
  • the invention has the following features:
  • the abalone body except the shell is used as the raw material for extraction.
  • the abalone's gastropods and organs can be used to extract polysaccharides, which can fully utilize resources and no waste discharge;
  • the supernatant was concentrated in vacuo to a polysaccharide content of 3% (detected by phenol sulfuric acid method); 3 times volume of 95% ethanol 21,000 ml was added to the concentrate, and the alcohol was precipitated at a temperature of 4 ° C for 16 hours, and centrifuged at high speed 10 In the minute or so, a centrifugal insoluble matter is obtained.
  • the insoluble matter was dried under vacuum at a temperature of 6 CTC to obtain a coarse abalone.
  • the phenolic acid was determined by a phenolic acid content of 8. 1%.
  • the insoluble matter was dried under vacuum at a temperature of 50 ° C to obtain 298 g of dried abalone crude polysaccharide. Crush to below 160 mesh with a pulverizer. It was detected by phenol sulfuric acid method and its polysaccharide content was 9.3%.
  • Abalone gastrope 1000 g was taken, and treated as in Example 1, ultrasonic treatment with 20 KHZ for 60 minutes, and centrifugation at high speed to obtain a centrifugation supernatant and a precipitate.
  • the precipitate was added to 25,000 g of water, ultrasonically treated with 27 KHZ for 20 minutes, and centrifuged at a high speed to obtain a centrifugation supernatant and a precipitate. The resulting supernatant was combined.
  • the precipitate was added to 25000 g of water, 0.5 mol/L of K0H solution was added thereto, and the ratio was adjusted to 8, and 150 g of trypsin (enzyme activity 2500 u / mg) was added thereto, and after enzymolysis for 2 hours at 50 Torr, The pH of 6 mol/L HC1 was adjusted to about 7 and the temperature was raised to 90 ° C for 5 minutes to purify the enzyme for 10 minutes, cooled to room temperature within 5 minutes, and centrifuged at high speed to obtain a supernatant and a precipitate. The resulting supernatant was combined. Concentrate, alcoholize, centrifuge as in Example 1. 5 ⁇ Drying abalone polysaccharide 43. 2 grams.
  • the resulting agglomerated polysaccharides were diluted with water to a specific gravity of 1. 07, spray drying, an inlet temperature of 160 ° C, an outlet temperature of 50 ⁇ , to obtain a dried abalone polysaccharide 43. 2 grams. It was detected by phenol sulfuric acid method and its polysaccharide content was 14.8%.
  • the obtained centrifuged insoluble matter was diluted with water to a specific gravity of 1.06, spray-dried, an inlet temperature of 160 Torr, and an outlet temperature of 50 ° C to obtain 236 g of dried abalone polysaccharide. 9% ⁇
  • the phenolic sulfuric acid method was determined to have a polysaccharide content of 13.9%.
  • Example 1 After enzymatic hydrolysis for 3 hours, the temperature was raised to 90-98 °C for 10 minutes in 2 to 5 minutes, and the mixture was cooled to room temperature in 5 minutes, and centrifuged to obtain a supernatant and a precipitate. The resulting supernatant was combined. Concentration, alcoholization, and centrifugation are as in Example 1. 5 ⁇ Drying abalone polysaccharide 225. 5 grams. The dried abalone polysaccharide 225. 5 grams. It was detected by a phenol sulfuric acid method and had a polysaccharide content of 14.2%.
  • the pH was adjusted to 9.0 with 0.5 mol/L of K0H solution, and 175 g of papain (enzyme activity: 1200 u / mg) was added, and the enzyme was stirred in a 35 ° C water bath. Solution for 3 hours, maintain the above pH value during hydrolysis; 'Re-use 6mol / L HC1 solution to adjust the pH to neutral, 5 minutes to 90 ° C to kill the enzyme for 10 minutes, 5 minutes to cool to room temperature, centrifuge to obtain the supernatant And precipitation; the resulting supernatant was combined. Concentrate, alcoholate, centrifuge as in Example 1.
  • the obtained centrifuged insoluble matter was diluted with water to a specific gravity of 1.1, spray-dried, an inlet temperature of 170 ° C, and an outlet temperature of 55 ° C to obtain 234 g of dried abalone polysaccharide.
  • the polysaccharide content was 15.2% as measured by a phenol sulfuric acid method.
  • a total of 1000 g of abalone was taken and treated as in Example 1.
  • the obtained slurry was irradiated with ultraviolet rays for 30 minutes, and autolyzed with NaCl at a concentration of 0.06 mol/l at pH 7.5, 50 ° C to obtain 6 mol/ from the solution.
  • L HC1 was adjusted to pH 5
  • 150 g of pepsin (enzyme activity 50 u/mg) was added, and the mixture was digested at 40 ° C for 2 hours.
  • the pH was adjusted to 7, 5 minutes, and the temperature was raised to 98 ° C for 10 minutes.
  • the mixture was cooled to room temperature in 5 minutes, and centrifuged at high speed to obtain a centrifugation supernatant and a precipitate.
  • Example 10 Take the whole abalone 1000g, treat it as in Example 1, and irradiate the obtained slurry with ultraviolet light for 10 minutes.
  • the NaCl having a degree of 0.08 mol/l was autolyzed at pH 7, 30 °C.
  • the rest of the operation is the same as in Example 10.
  • 50.9 g of dried abalone crude polysaccharide was obtained.
  • the polysaccharide content was determined by the phenol sulfuric acid method to be 14.4%.
  • the centrifugally insoluble matter is diluted with water to a specific gravity of 1.08, spray-dried, inlet temperature is 170, and the mixture is centrifuged. 1% ⁇ The phenolic sulphuric acid method was used to determine the polysaccharide content of 16.1%.
  • the abalone gastrope was taken in 1000 g, and the treatment was as in Example 1. Ultrasonic treatment was carried out for 30 minutes at 30 KHZ, and the supernatant and the insoluble matter were obtained by centrifugation. Add 25000 g of water to the insoluble matter, adjust the pH to 3 with 6 mol/L HCl, add 100 g of pepsin (enzyme activity 50 u/mg), stir and digest for 5 hours in a 40 ° C water bath, and maintain the above hydrolysis.
  • Example 17 pH value; adjust pH to 7 ⁇ 9 with 0.5mol/L K0H solution, add 80g trypsin, stir and digest for 2 hours in water bath at .55°C, keep the above pH value during hydrolysis; after enzymatic hydrolysis
  • the pH was adjusted to neutral with acid, the temperature was raised to 95 ° C for 5 minutes, the enzyme was incubated for 10 minutes, cooled to room temperature for 5 minutes, and centrifuged to obtain a supernatant and a precipitate. The supernatant separated by centrifugation was combined for use. Concentrate, alcoholate, centrifuge as in Example 1. 2 ⁇ Drying abalone polysaccharides 58. 2 grams.
  • the dried abalone polysaccharides were obtained by drying with water to a specific gravity of 1. 10, spray drying, an inlet temperature of 18 CTC, and an outlet temperature of 55 ° C.
  • the polysaccharide content was determined by the phenol sulfuric acid method to be 17.4%.
  • Example 17 Example 17:
  • AHP refined abalone organ polysaccharide
  • AHP was separated by Sephadex G-100 column chromatography. The eluent was 0.9% NaCl solution, the elution rate was 0.5 ml/min, and the phenol sulfuric acid method was used for tracking detection. Two elution peaks were drawn and the elution was collected separately. The peaks were dialyzed, dried, and then purified with DEAE cellulose 52 and eluted with a gradient of NaCl solution, each being a single eluting peak. Collect the pool, dialysis, and concentrate. Freeze-drying yielded uniform abalone organ polysaccharides AHP-1 and AHP-2.
  • AGP obtained in Example 24 dissolve it by adding 10 ml of water, and separate it by Sephadex G-100 column chromatography.
  • the eluent is 0.9% NaCl solution, elution rate is 0.5 ml/min, trace detection by phenol sulfuric acid method, draw a curve The only eluted peaks were collected.
  • AHP 100 obtained in Example 18 dissolve it by adding 10 ml of water, and separate it by Sephadex G-100 column chromatography.
  • the eluent is 0.9% NaCl solution
  • the elution rate is 0.5 ml/min
  • trace detection by phenol sulfuric acid method and curve is drawn.
  • Two eluting peaks were obtained, and the eluted peaks were collected, dialyzed, dried, and then purified by DEAE cellulose 52, and the gradient of the NaCl solution was a single eluting peak. Collect and combine, dialysis, and concentrate. Freeze-drying yielded uniform abalone organ polysaccharides AHP-1 and AHP-2.
  • Example 25 The AGPlOmg obtained in Example 25 was dissolved in ⁇ ultrapure water, sonicated for 20 minutes, filtered, and detected by gel chromatography. The molecular weight measured was 5,000-10,000 dal.
  • the gel column is Sepliarose CL-6B column, the eluent is 0.9% NaCl solution, flow rate 0.24 ml/min.
  • Example 26 10 mg of AHP-1 obtained in Example 26 was dissolved in 1 ml of ultrapure water, sonicated for 20 minutes, filtered, and detected by gel chromatography. The molecular weight measured was about 5 X 105 dal.
  • Example 26 The AHP-2 lOmg obtained in Example 26 was dissolved in 1 ml of ultrapure water, sonicated for 20 minutes, filtered, and detected by gel chromatography. The molecular weight measured was 10000-15000 dal.
  • the chromatographic conditions were: gas chromatograph (Agilent 6890N, USA), HP-1 column, stationary phase Methylsiloxane, carrier gas N2, flow rate 45 ml/min.
  • a hydrogen flame ionization detector (FID) was used to detect a temperature of 300 °C and an injection volume of 1 ⁇ l.
  • the inlet gasification temperature is 300 °C
  • the column temperature is programmed, 150 °C, lmin, 10 °C/min to 182 °C, 2 min, l °C/min to 188 °C, lmin, 8 °C/min To 230 ° C.
  • Example 26 Take AHP-1 obtained in Example 26, thoroughly dry to obtain 10 mg, add 2 ml of anhydrous HCI-methanol, fill with N2 tube, methyleneate at 80 ° C for 20 hours, take it out to room temperature, and neutralize with anhydrous KOH-methanol to The pH was 6, and then it was evaporated to dryness under reduced pressure at 40 ° C, and dried conventionally.
  • 0.2 ml of anhydrous pyridine was added to the thoroughly dried methanolysis product, dissolved at 75 ° C for 30 minutes, then 0.3 ml of silylating reagent was added, shaken, and allowed to stand for a few minutes. The supernatant was taken for gas chromatography analysis.
  • the chromatographic conditions were: gas chromatograph (Agilent 6890N, USA), HP-1 column, stationary phase Methylsiloxane, carrier gas N2, flow rate 45 ml/min.
  • the hydrogen flame ionization detector (FID) was used to detect the temperature at 300 ° C and the injection volume was 1 ⁇ .
  • the inlet gasification temperature is 300 °C
  • the column temperature is programmed, 150 °C, lmin, 10 °C/min to 182 °C, 2 min, rc/min To 188 ° C, lmin, 8. C/min to 230 °C.
  • Example 26 Take AHP-2 obtained in Example 26, thoroughly dry to obtain 10 mg, add 2 ml of anhydrous HC1-methanol, fill with N2 tube, methylene chloride at 80 ° C for 20 hours, take it out to room temperature, and neutralize with anhydrous KOH-methanol to The pH was 6, and then evaporated to dryness under reduced pressure at 40 ° C, and dried conventionally.
  • the chromatographic conditions were: gas chromatograph (Agilent 6890N, USA), HP-1 column, stationary phase Methylsiloxane, carrier gas N2, flow rate 45 nil/min. ⁇ Use hydrogen flame ionization detector (FID), the detection temperature is 300 °C, and the injection volume is 1 ⁇ 1.
  • the inlet gasification temperature is 300 °C
  • the column temperature is programmed, 150 °C, lmin, 10 °C/min to 182 °C, 2 min, VC/mm to 188 °C, lmin, 8 °C/min to 230 °C.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Animal Behavior & Ethology (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Polymers & Plastics (AREA)
  • Sustainable Development (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Materials Engineering (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Polysaccharides And Polysaccharide Derivatives (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Extraction Or Liquid Replacement (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)

Abstract

L'invention concerne un procédé d'extraction du polysaccharide d'ormeau. On obtient la chair d'ormeau (y compris les organes) en enlevant la coquille d'ormeau, en brisant la paroi à l'aide d'un hachoir, en la mélangeant unifomément avec de l'eau, en l'extrayant à une température comprise en 20 et 80 °C pendant 2 à 6 h, et en la séparant par centrifugation, la substance insoluble étant extraite de manière répétée une ou deux fois par une nouvelle addition d'eau, et le surnageant étant combiné. L'extrait est concentré à une teneur de saccharide de 1,5 à 3,0 %, précipité à une température comprise en 0 et 4 °C pendant 12 à 16 h par addition de 3 à 4 fois le volume de 95 % d'éthanol centrifugé, et séché par séchage sous vide ou séchage par pulvérisation pour obtenir le produit. Le polysaccharide d'ormeau uniforme est obtenu par purification avec sephadex G-100 ou DEAE-cellulose après évacuation de protéine et de glucogène. L'extraction peut également être réalisée par extraction alcaline, ultrasonique ou enzymatique. L'extraction d'enzyme peut être réalisée avec une enzyme simple, deux enzymes, une enzyme complexe ou leur combinaison. L'enzyme utilisée est la pepsine, la trypsase, etc. L'invention concerne un procédé permettant d'extraire de l'ormeau du polysaccharide (notamment des pléopodes et de ses organes). Ainsi, l'extraction et la séparation peuvent être réalisées efficacement. L'invention concerne également une base technique pour poursuivre la recherche, ce d'une manière continue et intensive en matière d'utilisation pharmaceutique du polysaccharide d'ormeau.
PCT/CN2006/002650 2005-10-11 2006-10-10 Procede d'extraction du polysaccharide de l'ormeau WO2007041951A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2008533850A JP4420470B2 (ja) 2005-10-11 2006-10-10 あわび多糖類抽出方法

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN200510047409.X 2005-10-11
CNB200510047409XA CN100336831C (zh) 2005-10-11 2005-10-11 鲍鱼多糖的提取方法

Publications (1)

Publication Number Publication Date
WO2007041951A1 true WO2007041951A1 (fr) 2007-04-19

Family

ID=36604952

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2006/002650 WO2007041951A1 (fr) 2005-10-11 2006-10-10 Procede d'extraction du polysaccharide de l'ormeau

Country Status (3)

Country Link
JP (1) JP4420470B2 (fr)
CN (1) CN100336831C (fr)
WO (1) WO2007041951A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102140141A (zh) * 2011-05-13 2011-08-03 陈锦权 鲍鱼多糖的制备工艺
CN111513269A (zh) * 2020-04-16 2020-08-11 自然资源部第三海洋研究所 一种鲍鱼内脏提取物及其应用

Families Citing this family (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101503721B (zh) * 2008-11-27 2012-01-11 浙江珍世堂生物科技有限公司 一种蚌肉多糖的提取方法
KR101257494B1 (ko) * 2010-05-27 2013-04-26 조선대학교산학협력단 전복 소화 가수분해물을 함유하는 면역계 염증성 질환 예방 및 치료용 조성물
CN101955548B (zh) * 2010-09-26 2012-05-23 陕西农产品加工技术研究院 酶法辅助提取蒺藜多糖的方法
CN102839149B (zh) * 2012-09-21 2013-11-13 中国海洋大学 鲍鱼内脏多糖替代血清在细胞培养基中的应用
CN103819575B (zh) * 2014-03-07 2016-03-02 潍坊医学院 一种土鳖虫多糖的提取方法、提取的多糖以及该多糖的用途
CN104381791B (zh) * 2014-11-25 2015-08-26 烟台新海水产食品有限公司 一种速溶鲍鱼覆盆子颗粒冲剂及其制作方法
CN104356249B (zh) * 2014-11-27 2016-07-13 厦门蓝湾科技有限公司 一种高纯鲍鱼多糖的制备工艺及其应用
CN104513843B (zh) * 2014-12-30 2018-01-19 集美大学 一种多糖和蛋白肽的联合制备方法
CN104497164A (zh) * 2015-01-26 2015-04-08 吉林大学 高压脉冲电场辅助酶解提取河蚌多糖工艺
CN105087713A (zh) * 2015-09-01 2015-11-25 集美大学 鲍鱼脏器多糖的制备方法
CN105272887B (zh) * 2015-10-20 2017-06-23 集美大学 一种从鲍鱼内脏中同时提取牛磺酸和多糖的方法
CN113150178B (zh) * 2021-04-14 2023-05-02 大连工业大学 一种硫酸化鲍鱼多糖及其在抑制新冠病毒中的应用
CN114133459A (zh) * 2021-11-23 2022-03-04 福建师范大学 一种鲍鱼多糖的制备方法及应用
CN114805623A (zh) * 2022-04-22 2022-07-29 福建大众健康生物科技有限公司 一种鲍鱼多糖的超低温冻融加压提取工艺
CN115025515B (zh) * 2022-06-10 2023-11-14 陈建 一种虎奶菇多糖提取分离设备
CN116693708B (zh) * 2023-04-28 2024-08-30 百珍堂生物科技(浙江)有限公司 一种高活性鲍鱼内脏多糖及其低共熔溶剂提取方法

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1473508A (zh) * 2003-08-07 2004-02-11 大连太平洋海龙科技有限公司 鲍鱼、海参、海胆营养液及其制造方法
CN1478423A (zh) * 2002-08-30 2004-03-03 大连轻工业学院 鲍鱼食品及其制备方法

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH10289760A (ja) * 1997-04-11 1998-10-27 Molex Inc 金属シェルの接続手段
CN1057776C (zh) * 1997-06-27 2000-10-25 吉林大学 人参、西洋参多糖皂甙联合提取工艺

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1478423A (zh) * 2002-08-30 2004-03-03 大连轻工业学院 鲍鱼食品及其制备方法
CN1473508A (zh) * 2003-08-07 2004-02-11 大连太平洋海龙科技有限公司 鲍鱼、海参、海胆营养液及其制造方法

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
"Extraction of Haliotis discushannai Ino. Polysaccharide and Experimental Study on Its Anti-tumor Action", CHINESE MODERN APPLIED PHARMACOLOGY, vol. 15, no. 1, February 1998 (1998-02-01) *
"Study on the Methanolysis of the Sulphated Polysaccharide Hal-A-from Haliotis diverisicolor Reeve", CHINESE JOURNAL OF ORGANIC CHEMISTRY, vol. 22, no. 5, 2002, pages 367 - 370, XP008080698 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102140141A (zh) * 2011-05-13 2011-08-03 陈锦权 鲍鱼多糖的制备工艺
CN111513269A (zh) * 2020-04-16 2020-08-11 自然资源部第三海洋研究所 一种鲍鱼内脏提取物及其应用
CN111513269B (zh) * 2020-04-16 2022-07-19 自然资源部第三海洋研究所 一种鲍鱼内脏提取物及其应用

Also Published As

Publication number Publication date
JP2009510234A (ja) 2009-03-12
CN100336831C (zh) 2007-09-12
JP4420470B2 (ja) 2010-02-24
CN1749280A (zh) 2006-03-22

Similar Documents

Publication Publication Date Title
WO2007041951A1 (fr) Procede d'extraction du polysaccharide de l'ormeau
Cheng et al. Extraction, characterisation and antioxidant activity of Allium sativum polysaccharide
CN101451157B (zh) 一种低分子量海参多糖的制备方法
CN112972488B (zh) 一种具有抗高尿酸血症活性的岩藻寡糖及其制备方法和应用
WO2017005134A1 (fr) Procédé de préparation et utilisation de polysaccharide de graine de lin présentant une activité antivirale et immunologique
CN107114793A (zh) 一种综合利用鲟鱼骨制备钙和硫酸软骨素的方法
CN115160449A (zh) 一种具有调节糖脂吸收作用的辣木叶多糖提取物及其制备方法与应用
JP4406032B2 (ja) 帆立貝多糖類抽出方法
CN114214366A (zh) 一种预防和治疗贫血的小肽粉和血红素之肽红复方药物及其制备方法和应用
CN112691115A (zh) 预防/治疗胃肠粘膜和肝脏损伤的阿拉伯木聚糖及复合物
CN113584108A (zh) 一种脾氨肽的制备方法
CN106749733B (zh) 一种泡叶藻硫酸酯化多糖及其制备方法、应用
CN109266703B (zh) 一种牡丹籽多糖酶解低聚糖的制备方法
JP5882635B2 (ja) マイタケ由来の高分子α−グルカン
JP4448330B2 (ja) マツタケ由来陰イオン交換樹脂吸着画分、免疫増強剤、及びストレス負荷回復促進剤
CN104725532B (zh) 一种精确定量控制肝素/类肝素中硫酸软骨素及硫酸皮肤素含量的方法
CN106589154B (zh) 一种野西瓜硒多糖的制备方法
CN114031695A (zh) 一种具有抗肿瘤功效的高纯度鲨鱼硫酸软骨素的制备工艺
JP2001327263A (ja) キノコ類の成分の抽出方法
CN102746413A (zh) 蜂花粉多糖酶解破壁结合热水超声浸提方法
JPH03227939A (ja) 免疫能賦活化物質及びその製造法
CN114671960B (zh) 玉米须多糖的制备方法和在降血糖产品中的应用
CN111778305B (zh) 一种用于提高免疫力的脾多肽组合物及口服液
CN116178575B (zh) 一种鸡枞菌多糖及其制备方法和应用
NL2031295B1 (en) Preparation method of pea protein tetrapeptide and its application in antihypertensive fields

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: 2008533850

Country of ref document: JP

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 06791230

Country of ref document: EP

Kind code of ref document: A1