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CN110551777A - preparation method of aloe polysaccharide - Google Patents

preparation method of aloe polysaccharide Download PDF

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Publication number
CN110551777A
CN110551777A CN201810538154.4A CN201810538154A CN110551777A CN 110551777 A CN110551777 A CN 110551777A CN 201810538154 A CN201810538154 A CN 201810538154A CN 110551777 A CN110551777 A CN 110551777A
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CN
China
Prior art keywords
aloe
polysaccharide
carrying
aloe vera
water
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Pending
Application number
CN201810538154.4A
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Chinese (zh)
Inventor
杨成东
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Nanjing Zelang Biotechnology Co Ltd
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Nanjing Zelang Biotechnology Co Ltd
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Priority to CN201810538154.4A priority Critical patent/CN110551777A/en
Publication of CN110551777A publication Critical patent/CN110551777A/en
Pending legal-status Critical Current

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    • CCHEMISTRY; METALLURGY
    • C08ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08BPOLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/0003General processes for their isolation or fractionation, e.g. purification or extraction from biomass
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00Preparation of compounds containing saccharide radicals
    • C12P19/04Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00Preparation of compounds containing saccharide radicals
    • C12P19/14Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biochemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Microbiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Sustainable Development (AREA)
  • Molecular Biology (AREA)
  • Materials Engineering (AREA)
  • Medicinal Chemistry (AREA)
  • Polymers & Plastics (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Polysaccharides And Polysaccharide Derivatives (AREA)

Abstract

The invention discloses a preparation method for extracting aloe polysaccharide from aloe vera, which is characterized by comprising the following steps of 1) taking dried aloe vera, crushing the dried aloe vera, sieving the crushed aloe vera with a 80-mesh sieve, collecting coarse powder for later use, 2) adding dried aloe vera powder into distilled water according to a water-material ratio of 5: 1 ~ 30: 1 (mL/g), rehydrating the mixture for 30min, adjusting the pH value to 7 ~ 9, 3) adding 0.1% ~ 0.3% of cellulase, carrying out ultrasonic extraction for 1 ~ 4h at the temperature of 50 ~ 110 ℃, 4) carrying out vacuum filtration on an extracting solution, taking a filtrate, carrying out vacuum rotary evaporation on the filtrate until the volume reaches 10mL, then adding 5 times of absolute ethyl alcohol by volume, uniformly mixing, placing the mixture in a refrigerator at 4 ℃ for 12h, centrifuging the mixture for 10min at 3000r/min to obtain crude polysaccharide, 5) putting the obtained crude polysaccharide on a macroporous adsorption resin column, carrying out distillation and elution by using 1 ~ 4 times of column volume of water, then carrying out elution by using 40 ~ 90% of ethyl alcohol, collecting an eluent, carrying out vacuum drying the reagent to obtain the aloe polysaccharide, and drying the aloe polysaccharide to obtain a high-quality aloe polysaccharide.

Description

Preparation method of aloe polysaccharide
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a preparation method of aloe polysaccharide.
Background
aloe (Aloe vera) is a perennial evergreen fleshy herbaceous plant of the genus Aloe of the family Liliaceae, distributed in tropical and subtropical regions, and currently has over 600 varieties, and is a widely used natural medicinal plant, wherein Aloe barbadensis, Aloe arborescens, Aloe china, and the like have important edible and medicinal values. Aloe is complex in chemical composition, and mainly contains anthraquinone substances, polysaccharides, amino acids, organic acids, vitamins, minerals and microelements. The aloe polysaccharide is the main bioactive substance in aloe gel, and has multiple effects of regulating immunity, resisting bacteria, viruses, tumors and oxidation, preventing aging, preventing and treating AIDS, preventing and treating diabetes, improving immunity, etc.
At present, the extraction method of aloe polysaccharide has been reported more at home and abroad, and mainly comprises hot water extraction, microwave-assisted extraction and ultrasonic-assisted extraction, while the supercritical fluid extraction method has less reports. There is no report on the extraction of aloe polysaccharides by an enzymatic method, nor on the extraction of aloe polysaccharides by combining an enzyme with ultrasonic waves. The method combines ultrasonic-assisted cellulase-macroporous resin extraction and purification technologies to prepare the high-purity aloe polysaccharide.
disclosure of Invention
The invention aims to provide an extraction method of aloe polysaccharide, and the aloe polysaccharide has wide medicinal value.
The invention is completed by the following technical scheme, and the specific content comprises the following steps: extraction method of aloe polysaccharide
1) Pulverizing dried Aloe barbadensis Miller, sieving with 80 mesh sieve, and collecting coarse powder;
2) adding Aloe dry powder into distilled water at water-to-material ratio of 5: 1 ~ 30: 1 (mL/g), rehydrating for 30min, and adjusting pH to 7 ~ 9;
3) Adding 0.1% ~ 0.3.3% cellulase, and ultrasonic extracting at 50 ~ 110 deg.C for 1 ~ 4 h;
4) Vacuum filtering the extractive solution, vacuum rotary evaporating the filtrate to 10mL, adding 5 times volume of anhydrous ethanol, mixing, standing in 4 deg.C refrigerator for 12 hr, and centrifuging at 3000r/min for 10min to obtain crude polysaccharide;
5) Loading the obtained crude polysaccharide on macroporous adsorbent resin column, eluting with 1 ~ 4 times of column volume of distilled water, eluting with 40 ~ 90% ethanol, and collecting ethanol eluate;
6) Recovering the reagent, and vacuum drying to obtain aloe polysaccharide.
The water-material ratio in the step 2) is 5: 1 ~ 30: 1 (mL/g).
Adding 0.1% ~ 0.3.3% cellulase in the step 3), and performing ultrasonic treatment at 50 ~ 110 ℃ for 1 ~ 4 h.
The macroporous adsorption resin in the step 5) is one of JAD-1000, ZJX and HD-8 types.
The present invention will be further described with reference to specific embodiments, but the scope of the invention as claimed is not limited to the following embodiments.
the specific implementation mode is as follows:
Example 1:
pulverizing dried Aloe barbadensis Miller, sieving with 80 mesh sieve, and collecting coarse powder; mixing aloe dry powder according to the water-material ratio of 5: 1 (mL/g) is added into distilled water, the mixture is rehydrated for 30min, and the pH is adjusted to 7; adding 0.1% cellulase, and performing ultrasonic extraction at 50 deg.C for 1 hr; vacuum filtering the extractive solution, vacuum rotary evaporating the filtrate to volume of 10mL, adding 5 times of anhydrous ethanol, mixing, standing in 4 deg.C refrigerator for 12 hr, and centrifuging at 3000r/min for 10min to obtain crude polysaccharide; loading the obtained crude polysaccharide on macroporous adsorbent resin column JAD-1000, eluting with 1 times of column volume of distilled water, eluting with 40% ethanol, and collecting ethanol eluate; recovering the reagent, and vacuum drying to obtain aloe polysaccharide.
Example 2:
Pulverizing dried Aloe barbadensis Miller, sieving with 80 mesh sieve, and collecting coarse powder; mixing aloe dry powder according to the water-material ratio of 20: 1 (mL/g) is added into distilled water, the mixture is rehydrated for 30min, and the pH is adjusted to 8; adding 0.2% cellulase, and performing ultrasonic extraction at 80 deg.C for 3 hr; vacuum filtering the extractive solution, vacuum rotary evaporating the filtrate to volume of 10mL, adding 5 times of anhydrous ethanol, mixing, standing in 4 deg.C refrigerator for 12 hr, and centrifuging at 3000r/min for 10min to obtain crude polysaccharide; loading the obtained crude polysaccharide on macroporous adsorbent resin column, eluting with 2 times of column volume of distilled water, eluting with 60% ethanol, and collecting ethanol eluate; recovering the reagent, and vacuum drying to obtain aloe polysaccharide.
example 3:
Pulverizing dried Aloe barbadensis Miller, sieving with 80 mesh sieve, and collecting coarse powder; mixing aloe dry powder according to the water-material ratio of 30: 1 (mL/g) is added into distilled water, the mixture is rehydrated for 30min, and the pH is adjusted to 9; adding 0.3% cellulase, and performing ultrasonic extraction at 110 deg.C for 4 hr; vacuum filtering the extractive solution, vacuum rotary evaporating the filtrate to volume of 10mL, adding 5 times of anhydrous ethanol, mixing, standing in 4 deg.C refrigerator for 12 hr, and centrifuging at 3000r/min for 10min to obtain crude polysaccharide; loading the obtained crude polysaccharide on macroporous adsorbent resin column, eluting with 4 times of column volume of distilled water, eluting with 90% ethanol, and collecting ethanol eluate; recovering the reagent, and vacuum drying to obtain aloe polysaccharide.

Claims (4)

1. A preparation method for extracting aloe polysaccharide from aloe vera is characterized by comprising the following steps:
1) Pulverizing dried Aloe barbadensis Miller, sieving with 80 mesh sieve, and collecting coarse powder;
2) adding Aloe dry powder into distilled water at water-to-material ratio of 5: 1 ~ 30: 1 (mL/g), rehydrating for 30min, and adjusting pH to 7 ~ 9;
3) Adding 0.1% ~ 0.3.3% cellulase, and ultrasonic extracting at 50 ~ 110 deg.C for 1 ~ 4 h;
4) vacuum filtering the extractive solution, vacuum rotary evaporating the filtrate to volume of 10mL, adding 5 times of anhydrous ethanol, mixing, standing in 4 deg.C refrigerator for 12 hr, and centrifuging at 3000r/min for 10min to obtain crude polysaccharide;
5) loading the obtained crude polysaccharide on macroporous adsorbent resin column, eluting with 1 ~ 4 times of column volume of distilled water, eluting with 40 ~ 90% ethanol, and collecting ethanol eluate;
6) Recovering the reagent, and vacuum drying to obtain aloe polysaccharide.
2. the method for extracting aloe polysaccharides from aloe vera according to claim 1, wherein the water to material ratio of step 2) is 5: 1 ~ 30: 1 (mL/g).
3. The method of claim 1, wherein the step 3) is carried out by adding 0.1% ~ 0.3.3% cellulase at 50 ~ 110 deg.C and under ultrasound for 1 ~ 4 h.
4. the method of claim 1, wherein the macroporous adsorbent resin of step 5) is one of JAD-1000, ZJX, and HD-8 types.
CN201810538154.4A 2018-05-30 2018-05-30 preparation method of aloe polysaccharide Pending CN110551777A (en)

Priority Applications (1)

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Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201810538154.4A CN110551777A (en) 2018-05-30 2018-05-30 preparation method of aloe polysaccharide

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111304266A (en) * 2019-12-12 2020-06-19 武汉新华扬生物股份有限公司 Biological enzymolysis fermentation process for aloe
CN114149512A (en) * 2020-09-08 2022-03-08 戴垄生技股份有限公司 Aloe extract and its preparation method

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111304266A (en) * 2019-12-12 2020-06-19 武汉新华扬生物股份有限公司 Biological enzymolysis fermentation process for aloe
CN114149512A (en) * 2020-09-08 2022-03-08 戴垄生技股份有限公司 Aloe extract and its preparation method
CN114149512B (en) * 2020-09-08 2023-11-14 戴垄生技股份有限公司 Aloe extract and its preparation method

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