Spirulina phatensis polysaccharide and extraction method thereof

A technology of spirulina polysaccharide and extraction method, which is applied in the field of deep processing of natural products, can solve the problems of extraction and purification technology staying in the laboratory research stage, cumbersome operation steps, toxicity, etc., and achieve good anti-oxidation and scavenging free radical effect, extraction process Simple, low-cost effect

Active Publication Date: 2015-01-28
SHANTOU UNIV
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  • Claims
  • Application Information

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Problems solved by technology

However, because the traditional hot water extraction method needs to consume a large amount of organic solvents to remove protein from Spirulina polysaccharides, and the operation steps are cumbersome, and at the same time, the activity of polysaccharides is also damaged to a certain extent, and further column chromatography purification of polysaccharides is even more limited. , making it difficult to separate and purify spirulina polysaccharides for industrial production
At present, the industrial production of spirulina in my country has been successful. The outstanding problems in the production and development process are: the deep-processing products of spirulina are in the primary stage, and domestic extraction and purification technologies of spirulina polysaccharides mostly stay in the laboratory research stage. Less production practice
The Sevage method is a classic method for removing protein, but it is usually only suitable for removing a small amount of protein and must be repeated many times; TCA method can shorten the process and reduce the loss rate of polysaccharides, which is beneficial to subsequent purification, but the loss of polysaccharide activity is serious; using enzymes Degrading protein by this method can increase the yield of polysaccharide crude product, but there is still more protein in the obtained composition
[0004] Moreover, the organic solvents used in these protein removal methods have certain toxicity, which will reduce the activity of polysaccharides to a certain extent in the process of protein removal, and the operation is cumbersome and troublesome; and the chromatography column used for further purification of spirulina polysaccharides obtains The amount of polysaccharide is limited. In addition, the packing medium of the chromatographic column also has defects such as expensive price and limited number of times of use, which limits the industrial separation and purification of spirulina polysaccharide.

Method used

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  • Spirulina phatensis polysaccharide and extraction method thereof
  • Spirulina phatensis polysaccharide and extraction method thereof
  • Spirulina phatensis polysaccharide and extraction method thereof

Examples

Experimental program
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Embodiment 1

[0035] Preparation of Spirulina Polysaccharide Extract

[0036] The concrete process of preparing Spirulina polysaccharide extract in the present embodiment is as follows: figure 1 shown.

[0037] (1) Preparation of spirulina powder suspension: 8 g of Spirulina platensis powder was weighed and dissolved in 400 mL of phosphate buffer (0.01 mol / L, pH=7) to prepare a spirulina powder suspension.

[0038] (2) Preparation of spirulina cell disruption solution 1: put the spirulina powder suspension at 4°C for 4 hours, then freeze it at -20°C for 3 hours, crush the ice cubes, and put it at 4°C for 4 hours. Freeze at -20°C for 3 hours, crush the ice cubes, repeat this three times, and centrifuge at 6000 rpm for 20 minutes to obtain Spirulina cell disruption solution 1 and cell fragments;

[0039] (3) Preparation of spirulina cell disruption solution 2: add ammonium sulfate solid to spirulina cell disruption solution 1 until its saturation is 50%, stir it with a magnetic stirrer to c...

Embodiment 2

[0050] Evaluation of Antioxidant Activity Determination Index

[0051] The sample solution in this embodiment is the polysaccharide extract in different extraction steps.

[0052] DPPH system: refer to the methods of Capek.P and others, and use ascorbic acid (Vc) as a positive control to measure the scavenging rate of DPPH free radicals. The samples or Vc were dissolved in water to prepare 6 gradient samples or Vc solutions (20, 40, 80, 160, 320, 640 μg / mL). According to Table 1, add 2mL of the above solution (not added to the blank tube) and 2mL DPPH solution (not added to the background tube) into the test tube in turn, make up to 4mL in each tube with distilled water, mix with a shaker, let it stand at room temperature for 30min, and measure at 517nm with Distilled water was adjusted to zero to determine the absorbance, and the calculation formula was:

[0053]

[0054] Where: V DPPH is the absorbance of the DPPH solution without adding sample or Vc; A 实验 Absorbance ...

Embodiment 3

[0070] Determination of structure by infrared spectroscopy

[0071] Determination of infrared spectrogram: dialyze the obtained lower phase spirulina polysaccharide and the spirulina polysaccharide solution after passing through the column to desalinate and then freeze-dry to obtain spirulina polysaccharide powder. Press with KBr, use infrared spectrometer at 400-4000cm-1 range scan.

[0072] Spirulina polysaccharide solution through Sephadex G-150 gel column chromatography steps:

[0073] (1) Pretreatment: Weigh about 5 g of Sephadex G-150, add 100 ml of distilled water, and let it swell at room temperature for 3 hours.

[0074] (2) Column packing: the diameter and column length of the gel chromatography column are 1.6×40cm. The bottom of the column was plugged tightly with a rubber stopper equipped with a thin glass tube, and the bottom was covered with a clean nylon cloth. Then install the column vertically, first add 1 / 3 column volume of distilled water, then continuous...

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Abstract

The invention relates to a spirulina phatensis polysaccharide and an extraction method thereof. The extraction method comprises the following steps: after multigelation and wall breaking of a spirulina phatensis powder suspension, centrifuging to obtain a cell lysate 1 and cell debris; adding ammonium sulfate in the cell lysate 1 until the saturation is 50%, after salting-out precipitation, centrifuging to remove phycobiliprotein to obtain a supernatant, and evenly mixing the supernatant and the cell debris to obtain a cell lysate 2; obtaining a spirulina phatensis polysaccharide crude extract by using a hot water extraction method; adding the crude extract to an ethanol/ammonium sulfate aqueous two-phase system, and extracting to obtain a bottom-phase solution of spirulina phatensis polysaccharide with higher purity; after the bottom-phase solution is desalted by dialysis, eluting by using a Sephadex G-150 chromatographic column and a DEAE Sephadex A-50 anion exchange column to obtain a pure spirulina phatensis polysaccharide solution; and freeze-drying, thereby obtaining a pure spirulina phatensis polysaccharide finished product. The extraction method can be used for avoiding the traditional complicated protein removal operation steps, has low cost, high yield, and high purity and activity of polysaccharide, and is suitable for intermittent and large-scale production and processing.

Description

technical field [0001] The invention relates to the technical field of deep processing of natural products, in particular to a spirulina polysaccharide and an extraction method thereof. Background technique [0002] Spirulina contains a variety of biologically active substances, and the spirulina polysaccharide (content is about 2.0% to 3.0%) is a water-soluble acidic heteropolysaccharide, which is extracted and separated from spirulina algae and spirulina culture fluid A class of biologically active substances in the form of white powder. Spirulina polysaccharide is another biologically active substance with anti-tumor, anti-radiation and anti-mutation functions in addition to phycobiliprotein in Spirulina. It can improve the body's cellular and humoral immune functions, resist cancer cell proliferation, and reduce radiation-induced Genetic damage, etc.; can be used for anti-cancer, anti-aging, enhance the body's immunity and so on. However, because the traditional hot wa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/00
Inventor 刘杨李瑞昌颜晓琳肖湘
Owner SHANTOU UNIV
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