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Method for eutectic solvent and salt double aqueous phase extraction of phycocyanin from spirulina

A technology of deep eutectic solvent and phycocyanin, which is applied to the preparation method of peptides, chemical instruments and methods, and peptide decapitation, can solve the problems of complex synthesis, high cost, and toxicity, and achieve high yield and purity, Ease of operation and mild conditions

Pending Publication Date: 2017-07-11
HEXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

CN105820236A、CN102993297A、CN104672325A、CN106008705A、CN105820238A、CN105820237A、CN104479009A、CN104387468A和CN104387469A均采用聚合物 / 盐双水相体系萃取螺旋藻藻蓝蛋白;CN103880950A公开了一种离子液体双水相体系提取藻蓝蛋白的 method, as a new type of green solvent and functional medium material, ionic liquid has the characteristics of low vapor pressure, non-volatility, strong solubility and good chemical stability, and has become a substitute for traditional organic solvents; but its synthesis is complicated , high cost, and the imidazole and pyridine ionic liquids themselves have the defects of being difficult to degrade and be toxic; and the emergence of deep eutectic solvents not only inherits the excellent characteristics of ionic liquids, but also overcomes its disadvantages, and has the advantages of simple synthesis , low cost, safety, biocompatibility and other characteristics, it is a new type of solvent with the concept of green chemistry
However, there is no report on the separation of spirulina phycocyanin by using deep eutectic solvent two-phase extraction

Method used

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  • Method for eutectic solvent and salt double aqueous phase extraction of phycocyanin from spirulina

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A method for deep eutectic solvent-salt two-phase extraction of phycocyanin in spirulina, comprising the steps:

[0029] S1: Ultrasonic-assisted deep eutectic solvent extraction: Add 55 g of deep eutectic solvent made of tetramethylammonium chloride and urea with a molar ratio of 1:1 to 1 g of spirulina powder, and after 100 W ultrasonic treatment for 5 min, the Centrifuge at 5000r / min for 5min to obtain a crude extract containing phycocyanin;

[0030] S2: deep eutectic solvent-salt aqueous two-phase extraction: add K to the crude extract obtained in step S1 2 HPO 4 and deionized water to make K 2 HPO 4 The mass fraction of the crude extract is 35%, and the mass fraction of the crude extract is 25%. Mix well to obtain a two-phase system, adjust the pH to 6.0 and mix well, let stand at room temperature for 10min to separate layers, and obtain an upper phase and a lower phase;

[0031] S3: secondary aqueous two-phase extraction: take the upper phase separated in step ...

Embodiment 2

[0034] A method for deep eutectic solvent-salt two-phase extraction of phycocyanin in spirulina, comprising the steps:

[0035] S1: Ultrasonic-assisted deep eutectic solvent extraction: Add 90 g of deep eutectic solvent made of betaine and glycerol with a molar ratio of 1:2 to 1.5 g of spirulina powder, and after 10 min of ultrasonic treatment at 75 W, at 4000 r / min Centrifuge for 5 minutes under the same conditions to obtain a crude extract containing phycocyanin;

[0036] S2: deep eutectic solvent-salt aqueous two-phase extraction: add K to the crude extract obtained in step S1 2 HPO 4 and deionized water to make K 2 HPO 4 The mass fraction of the crude extract is 38%, and the mass fraction of the crude extract is 27%. Mix well to obtain a two-phase aqueous system, adjust the pH to 6.0 and mix well, let stand at room temperature for 12 minutes to separate layers, and obtain an upper phase and a lower phase;

[0037] S3: secondary aqueous two-phase extraction: take the up...

Embodiment 3

[0040] A method for deep eutectic solvent-salt two-phase extraction of phycocyanin in spirulina, comprising the steps:

[0041]S1: Ultrasonic-assisted deep eutectic solvent extraction: Add 70 g of deep eutectic solvent made of choline chloride, glucose and ethylene glycol with a molar ratio of 1:1:2 to 1 g of spirulina powder, and treat with 125W ultrasonic After 8 minutes, centrifuge at 6000r / min for 3 minutes to obtain a crude extract containing phycocyanin;

[0042] S2: deep eutectic solvent-salt aqueous two-phase extraction: add K to the crude extract obtained in step S1 2 HPO 4 and deionized water to make K 2 HPO 4 The mass fraction of the crude extract is 25%, and the mass fraction of the crude extract is 25%. Mix well to obtain a two-phase system, adjust the pH to 7.0 and mix well, let stand at room temperature for 15 minutes to separate layers, and obtain an upper phase and a lower phase;

[0043] S3: secondary aqueous two-phase extraction: take the upper phase sep...

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Abstract

The invention provides a method for eutectic solvent and salt double aqueous phase extraction of phycocyanin from spirulina. The method comprises steps of supersonic-wave-assisted eutectic solvent extraction, eutectic solvent and salt double aqueous phase extraction, secondary double aqueous phase extraction and recycle of the eutectic solvent and phycocyanin. According to the method, the recyclable eutectic solvent is taken as an extraction solvent and forms a double aqueous phase system together with salt for separation and purification. The advantages of the eutectic solvent and double aqueous phase are combined, so that the method is simple to operate and saved in time, the yield and purity of phycocyanin are obviously improved, and the method is suitable for massive industrial production.

Description

technical field [0001] The invention belongs to the technical field of separation and purification of active ingredients of natural products, and particularly relates to a method for extracting phycocyanin in spirulina with a deep eutectic solvent-salt two-phase aqueous phase. Background technique [0002] Phycocyanin, also known as phycocyanin, is a dark blue powder isolated from spirulina. Phycocyanin is divided into C-phycocyanin and R-phycocyanin, the former is mainly found in cyanobacteria, the latter is mainly found in red algae, and both are found in cryptophycobacteria. Phycocyanin is not only a good food coloring, but also a high-quality health food because of its complete range of amino acids and high content of essential amino acids. At the same time, it is often used as a fluorescent agent in experiments. [0003] The current methods for protein purification mainly include salting out precipitation, electrophoresis, ion exchange and adsorption, etc., but there a...

Claims

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Application Information

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IPC IPC(8): C07K14/795C07K1/14
CPCC07K14/795
Inventor 罗光宏张喜峰杨生辉王丹霞陈天仁祖廷勋
Owner HEXI UNIV
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