CN109134680B - Extraction and purification method of fucoidin and application thereof - Google Patents

Extraction and purification method of fucoidin and application thereof Download PDF

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CN109134680B
CN109134680B CN201811051757.8A CN201811051757A CN109134680B CN 109134680 B CN109134680 B CN 109134680B CN 201811051757 A CN201811051757 A CN 201811051757A CN 109134680 B CN109134680 B CN 109134680B
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extraction
fucoidan
filtering
purification
raw materials
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CN109134680A (en
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刘�东
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Shenzhen Huachuang Biomedical Technology Co ltd
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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
    • C08B37/006Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
    • C08B37/0063Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/52Adding ingredients
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105Plant extracts, their artificial duplicates or their derivatives
    • 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
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00Food compositions, function of food ingredients or processes for food or foodstuffs

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Polymers & Plastics (AREA)
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  • Medicinal Chemistry (AREA)
  • Food Science & Technology (AREA)
  • Organic Chemistry (AREA)
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  • General Health & Medical Sciences (AREA)
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  • Polysaccharides And Polysaccharide Derivatives (AREA)
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Abstract

The invention belongs to the technical field of polysaccharide extraction, and particularly relates to an extraction and purification method of fucoidan and application thereof. The method comprises three steps of crude extraction, heavy metal removal and purification. The extracted fucoidin can be processed into health food. According to the technical scheme provided by the invention, the product which is further used for food processing and has high content and no heavy metal residue is obtained through three steps of crude extraction, heavy metal removal and purification in sequence, and the three steps are organically linked, so that the extraction process is greatly simplified, the use amount of water and a solvent is greatly reduced, two times of ultrasound are carried out, the extraction efficiency is facilitated, the heavy metal residue is removed, and the high-quality product is obtained.

Description

Extraction and purification method of fucoidin and application thereof
Technical Field
The invention belongs to the technical field of polysaccharide extraction, and particularly relates to an extraction and purification method of fucoidan and application thereof.
Background
The fucoidan component is limited to only the slimy part extracted from brown algae, and there are many different fucoidans, such as those extracted from kelp, undaria pinnatifida sporophyll and nemacystus, etc. According to recent studies, many biological activities of fucoidan have been disclosed, such as "anticancer effect", "cholesterol-lowering", "blood pressure-lowering" and "antiviral" effects. In the structure of chemical molecule, fucoidan is a high molecular polysaccharide composed of fucose sulfate, and besides, the sugar chain of fucoidan also includes galactose, mannose, xylose and glyconic acid.
The fucoidan extraction method mainly comprises an acid extraction method, a water extraction method and a salt extraction method, and the newly developed extraction method mainly comprises an ultrasonic wave extraction method, a microwave-assisted extraction method and an enzyme-assisted extraction method. The extraction and separation method of fucoidan is particularly critical because the activity of the fucoidan is related to the molecular weight, monosaccharide components, sulfate content and sulfate ester group composition, and sulfate loss and target compound structure transformation are easily caused in the separation process.
In addition, as the raw materials for extracting the fucoidin are marine products, the fucoidin has strong adsorbability to heavy metal ions due to inevitable pollution of seawater, so that some heavy metal ions can be remained in the extracted fucoidin, and troubles are caused for subsequent use of health-care food or medicines.
Disclosure of Invention
The invention aims to provide a method for extracting and purifying fucoidan and application thereof.
In order to solve the technical problems, the technical scheme of the invention is as follows: the extraction and purification method of fucoidin comprises the following steps:
step one, crude extraction: crushing the raw materials, and then obtaining an immersion liquid of fucoidin by adopting an acid extraction method and/or a hot water extraction method;
step two, removing heavy metals: adding a sulfhydryl chelating agent into the steeping liquor, heating and ultrasonically vibrating, and then precipitating by using absolute ethyl alcohol to obtain a filter cake of fucoidin;
step three, purification: the fucoidin is further purified by absolute ethyl alcohol precipitation, quaternary ammonium salt precipitation or solvent extraction.
Optionally, the raw material is one or more of kelp, undaria pinnatifida sporophyll and nemacystus decipiens.
Optionally, the specific process of the step one is as follows: crushing the dried raw materials by a crusher, and soaking the crushed raw materials in 200mL of acetic acid aqueous solution with the pH value of 4-5 for 2-3 hours; filtering, retaining filtrate, further grinding filter cake into slurry, adding water 8-12 times of the weight of the raw material, heating to 30-40 deg.C, ultrasonic extracting for 30-60min, and filtering; the two filtrates are combined and adjusted to pH 6.5-7.5.
Optionally, the sulfhydryl chelator of step two is selected from 1 to 3g of a sulfhydryl chelator (2, 3-dimercaptopropanol-1-sulfonic acid or 2, 3-dimercaptosuccinic acid).
Optionally, the mercapto-chelating agent is used in an amount of 1-3% by weight of the starting material.
Optionally, the specific process of the second step is as follows: adding a sulfhydryl chelator into the solution obtained in the first step, heating to 40-50 ℃, ultrasonically vibrating for 30-60min, adjusting pH value to 6.5-7.5, adding anhydrous ethanol 8-12 times of the weight of the raw material, standing overnight, and filtering.
Optionally, the specific process of the step three is as follows: and (3) dissolving the filter cake obtained in the second step by using a calcium chloride aqueous solution, adding equal volume of absolute ethyl alcohol to precipitate the fucoidan, filtering, washing for 3 times by using the absolute ethyl alcohol, and drying to obtain an extracted product.
Optionally, the concentration of calcium chloride in the calcium chloride aqueous solution is 5-10% wt.
The invention also provides the application of the fucoidin prepared by the purification and purification method in health-care functional food.
Optionally, the food product is a beverage or a brewable powder.
According to the technical scheme provided by the invention, the product which is further used for food processing and has high content and no heavy metal residue is obtained through three steps of crude extraction, heavy metal removal and purification in sequence, and the three steps are organically linked, so that the extraction process is greatly simplified, the use amount of water and a solvent is greatly reduced, two times of ultrasound are carried out, the extraction efficiency is facilitated, the heavy metal residue is removed, and the high-quality product is obtained.
Detailed Description
For the convenience of understanding, the process for extracting and purifying fucoidan is described in the following with reference to examples, which are intended to illustrate the present invention and not to limit the scope of the present invention.
The raw materials used in the examples are all conventional commercial products except for special instructions, and the processing techniques used are all the techniques common in the art except for special instructions.
Example 1
The extraction and purification method of fucoidin comprises the following steps:
step one, crude extraction: crushing dried 100g of raw material (kelp) by a crusher, and soaking the crushed raw material in 200mL of acetic acid aqueous solution with the pH value of 4 for 3 hours; filtering, retaining filtrate, further grinding filter cake into slurry, adding 1L water, heating to 40 deg.C, ultrasonic extracting for 30min, and filtering; combining the two filtrates, and adjusting the pH value to about 7;
step two, removing heavy metals: adding 1g of sulfhydryl chelator (2, 3-dimercaptopropanol-1-sulfonic acid) into the solution obtained in the first step, heating to 50 ℃, ultrasonically vibrating for 60min, adjusting the pH value to about 7 again, adding 1L of absolute ethyl alcohol, standing overnight, and filtering;
step three, purification: and (3) dissolving the filter cake obtained in the second step by using a 10% wt calcium chloride aqueous solution, adding equal volume of absolute ethyl alcohol to precipitate the fucoidan, filtering, washing for 3 times by using the absolute ethyl alcohol, and drying to obtain an extracted product.
Example 2
The extraction and purification method of fucoidin comprises the following steps:
step one, crude extraction: sun-drying 100g of raw material (Nemacystus decipiens), pulverizing with a pulverizer, and soaking in 200mL of acetic acid aqueous solution with pH of 5 for 3 hours; filtering, retaining filtrate, further grinding filter cake into slurry, adding 1L water, heating to 30 deg.C, ultrasonic extracting for 60min, and filtering; combining the two filtrates, and adjusting the pH value to about 7;
step two, removing heavy metals: adding 3g of sulfhydryl chelator (2, 3-dimercaptosuccinic acid) into the solution obtained in the first step, heating to 40 ℃, ultrasonically vibrating for 30min, adjusting the pH value to about 7 again, adding 1L of absolute ethyl alcohol, standing overnight, and filtering;
step three, purification: and (3) dissolving the filter cake obtained in the second step by using 8% wt of calcium chloride aqueous solution, adding equal volume of absolute ethyl alcohol to precipitate the fucoidan, filtering, washing for 3 times by using the absolute ethyl alcohol, and drying to obtain an extracted product.
Example 3
The extraction and purification method of fucoidin comprises the following steps:
step one, crude extraction: sun-drying 100g of raw materials (sea tangle, Undaria pinnatifida sporophyll and Nemacystus decipiens and mixture), pulverizing with a pulverizer, and soaking in 200mL of acetic acid aqueous solution with pH of 4 for 2 hours; filtering, retaining filtrate, further grinding filter cake into slurry, adding 1L water, heating to 30 deg.C, ultrasonic extracting for 50min, and filtering; combining the two filtrates, and adjusting the pH value to 6.5;
step two, removing heavy metals: adding 2g of heavy metal sulfydryl chelating agent (2, 3-dimercaptopropanol-1-sulfonic acid) into the solution obtained in the step one, heating to 40 ℃, ultrasonically vibrating for 40min, adjusting the pH value to 6.5 again, adding 1L of absolute ethyl alcohol, standing overnight, and filtering;
step three, purification: and (3) dissolving the filter cake obtained in the second step by using a 6% wt calcium chloride aqueous solution, adding equal volume of absolute ethyl alcohol to precipitate the fucoidan, filtering, washing for 3 times by using the absolute ethyl alcohol, and drying to obtain an extracted product.
Comparative example 1
Extracting with common hot water, and purifying by precipitating with anhydrous ethanol for 2-3 times.
Experimental data
The examples 1-3 and comparative examples were subjected to compositional measurements and the specific data are shown in table 1.
TABLE 1
Composition (I) Raw material Example 1 Example 2 Example 3 Comparative example
Total polysaccharide (%) 25.1 83.9 81.6 80.4 75.3
Sulfates of sulfuric acid 1.25 35.8 38.7 32.8 27.3
As(mg/kg) 1.61 Not detected out Not detected out Not detected out 1.35
Pb(mg/kg) 1.23 Not detected out Not detected out Not detected out 0.84
Wherein the detection method of total polysaccharide is phenol-cysteine-sulfuric acid method, the detection method of sulfate is gelatin-barium chloride method, the detection method of As is described in GB/T5009.11-2003, and the detection method of Pb is described in GB/T5009.11-2003
GB5009.12-2010。
As can be seen from Table 1, the fucoidan extracted by the technical method provided by the invention has the advantages of minimal loss of active sulfate and complete removal of residual heavy metals.
Finally, it should be noted that: the above examples are only for illustrating the technical solutions of the present invention, and are not limited thereto. Although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: it is to be understood that modifications may be made to the technical solutions described in the foregoing embodiments, or some or all of the technical features may be equivalently replaced, and such modifications or replacements may not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.

Claims (5)

1. The extraction and purification method of fucoidin is characterized by comprising the following steps:
step one, crude extraction: crushing the dried raw materials by a crusher, and soaking the crushed raw materials in 200mL of acetic acid aqueous solution with the pH value of 4-5 for 2-3 hours; filtering, retaining filtrate, further grinding filter cake into slurry, adding water 8-12 times of the weight of the raw material, heating to 30-40 deg.C, ultrasonic extracting for 30-60min, and filtering; mixing the two filtrates, and adjusting pH to 6.5-7.5;
step two, removing heavy metals: adding a sulfhydryl chelator into the filtrate obtained in the step one, wherein the sulfhydryl chelator is selected from 2, 3-dimercaptopropanol-1-sulfonic acid or 2, 3-dimercaptosuccinic acid, the dosage of the sulfhydryl chelator is 1-3% of the weight of the raw materials, heating to 40-50 ℃, ultrasonically vibrating for 30-60min, adjusting the pH value to 6.5-7.5 again, adding anhydrous ethanol with the weight 8-12 times that of the raw materials, standing overnight, and filtering;
step three, purification: and (3) dissolving the filter cake obtained in the second step by using a calcium chloride aqueous solution, adding equal volume of absolute ethyl alcohol to precipitate the fucoidan, filtering, washing for 3 times by using the absolute ethyl alcohol, and drying to obtain the extracted product.
2. The method for extracting and purifying fucoidan according to claim 1, wherein the raw material is one or more of kelp, sporophyll of undaria pinnatifida, and nemacystus decipiens.
3. The extraction and purification method of fucoidan according to claim 1, wherein the concentration of calcium chloride in the calcium chloride aqueous solution is 5-10% wt.
4. The use of fucoidan prepared by the extraction and purification method according to any one of claims 1 to 3 for preparing a health functional food.
5. Use according to claim 4, wherein the foodstuff is a beverage or an infusible powder.
CN201811051757.8A 2018-09-10 2018-09-10 Extraction and purification method of fucoidin and application thereof Expired - Fee Related CN109134680B (en)

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RU2707872C1 (en) * 2019-05-22 2019-12-02 федеральное государственное автономное образовательное учреждение высшего образования "Южно-Уральский государственный университет (национальный исследовательский университет)" Micronization method of fucoidan
CN110981988B (en) * 2019-12-25 2022-05-31 无锡先君生物医药有限公司 Extraction method and application of fucoidin
CN113278083A (en) * 2020-05-19 2021-08-20 杭州鹿扬科技有限公司 Low molecular weight polysaccharides with antithrombotic activity
EP4302772A1 (en) 2022-07-06 2024-01-10 Müller, Jan Allan Composition for oral administration and supplement comprising such a composition

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