Technology for extracting chlorophyll from spirulina

A technology of spirulina and chlorophyll, applied in the direction of organic chemistry, can solve the problems of reducing yield and biological activity, large solvent consumption, chlorophyll decomposition, etc., to improve food safety, improve yield and purity, and promote the reaction. Effect

Active Publication Date: 2019-08-23
北海生巴达生物科技有限公司
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AI-Extracted Technical Summary

Problems solved by technology

The commonly used organic solvent extraction method is time-consuming, consumes a lot of solvents, is highly toxic, destroys nutrients, is costly, and is easy to decompose chlorophyll at high temperatures, thereby reducing yield and biological activity.
At present, although there are relevant literature reports on extracting chlorophyll from Spirulina, the comprehensive development and utilization technology of Spirulina and other biomass in the prior art is still immature, and most of Spirulina and other biomass have not been fully utilized, resulting in biomass A huge waste of energy; and, because th...
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Abstract

The invention discloses a technology for extracting chlorophyll from spirulina. The technology for extracting the chlorophyll from the spirulina comprises the steps of raw material pre-treatment, soaking, enzymolysis, filtering and drying. According to the technology for extracting the chlorophyll from the spirulina, when the chlorophyll is extracted from the spirulina, modes of soaking, enzymolysis, microwaves and ultrasonic treatment are adopted for obtaining the chlorophyll, and use of organic solvents is avoided, so that the food safety of the chlorophyll is improved. Enzymes of protease,cellulase, pectinase, lignase, amylase, esterase and the like are adopted for conducting enzymolysis treatment on the spirulina, substances other than the chlorophyll in the spirulina are degraded, the chlorophyll in the spirulina is not destructed, thus the chlorophyll contained in the spirulina is extracted as much as possible, and a product does not contain other impurities, so that the yield and the purity of the chlorophyll are improved, the extraction yield of the chlorophyll is higher than 99.5%, and is higher than that of a traditional solvent extraction method by 8.8% or above, and the purity of the chlorophyll is higher than 99.96%.

Application Domain

Technology Topic

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  • Technology for extracting chlorophyll from spirulina

Examples

  • Experimental program(5)
  • Comparison scheme(3)
  • Effect test(1)

Example Embodiment

[0039] Example 1
[0040] A process for extracting chlorophyll from spirulina, comprising the following steps:
[0041] (1) Raw material pretreatment: Wash the fresh spirulina, treat it in the dark at 1°C for 80 minutes, then pulverize it at 8°C until it passes through a 50-mesh sieve, put it into a centrifuge tube equipped with steel balls, and pour it into liquid nitrogen. When the liquid nitrogen has just evaporated, cover the centrifuge tube and place it on the vortexer to vortex, and vortex to the spirulina powder that has passed a 250-mesh sieve; during the vortexing process, the centrifuge tube is repeatedly inverted 3 times;
[0042] (2) immersion: under dark and argon or nitrogen protection conditions, add deionized water immersion treatment to the above-mentioned spirulina powder, and adopt microwave and ultrasonic treatment to assist immersion to obtain a spirulina aqueous solution;
[0043] The microwave and ultrasonic treatment-assisted soaking methods are as follows: first, microwave treatment is performed at a microwave power of 50W and a microwave temperature of 2° C. for 45 s, and then at an ultrasonic power of 105W and an ultrasonic temperature of 18° C. for 3 min. The microwave and ultrasonic treatments were alternately performed twice; the soaking parameters were controlled as follows: the soaking pressure was 0.05MPa, the soaking temperature was 1°C, and the soaking time was 2.5h;
[0044] (3) Enzymatic hydrolysis: under dark and argon or nitrogen protection conditions, add mixed enzyme A to the above-mentioned Spirulina aqueous solution for enzymatic hydrolysis, and use microwave treatment to assist enzymatic hydrolysis, then add mixed enzyme B, and further use ultrasonic treatment to assist enzyme solution to obtain an enzymatic hydrolysis mixture;
[0045] Described microwave treatment: microwave power is 85W, microwave time is 60s, microwave temperature is 2 ℃; Described ultrasonic treatment: ultrasonic power is 105W, ultrasonic wave time is 3min, ultrasonic temperature is 18 ℃; Described mixed enzyme A is protease, Cellulase, pectinase, hemicellulase, and peroxidase are in a mass ratio = 3:1:1:0.3:0.1; before adding mixed enzyme B, the temperature is raised to 40°C and an enzyme inhibitor is added to adjust the pH. Inactivate the enzyme, cool down to 1 °C and then add mixed enzyme B; after the ultrasonic treatment is completed to assist the enzymatic hydrolysis, the material is heated to 42 °C and an enzyme inhibitor is added to adjust the pH to inactivate the enzyme, and then the temperature is lowered to 8 °C to obtain the enzymatic hydrolysis mixture; Mixed enzyme B is lignase, cellulase, amylase, esterase, lipoxygenase and pectinase according to the mass ratio=1:2:1:0.5:0.5:1;
[0046] (4) Filtration: under dark and argon or nitrogen protection conditions, use an ultrafiltration membrane to filter the above-mentioned enzymatic hydrolysis mixture, remove the filtration mother liquor, and obtain a chlorophyll semi-finished product;
[0047] (5) drying: under dark and argon or nitrogen protection conditions, the chlorophyll semi-finished product obtained above is sent into a dryer for drying, sealed, packaged, and protected from light to obtain the chlorophyll; the decompression drying The temperature is 26°C, the relative air humidity is less than or equal to 65%, the drying time is 22min, and the drying is performed until the chlorophyll water content is lower than 20%.

Example Embodiment

[0048] Example 2
[0049] A process for extracting chlorophyll from spirulina, comprising the following steps:
[0050] (1) Raw material pretreatment: Wash the fresh spirulina, treat it in the dark at 9°C for 90min, then pulverize it at 1°C until it passes through a 100-mesh sieve, put it into a centrifuge tube with steel balls, and pour it into liquid nitrogen. When the liquid nitrogen has just evaporated, cover the centrifuge tube and place it on the vortexer for vortexing, and vortex to the spirulina powder that has passed through a 350-mesh sieve; during the vortexing process, the centrifuge tube is repeatedly inverted 5 times;
[0051] (2) immersion: under dark and argon or nitrogen protection conditions, add deionized water immersion treatment to the above-mentioned spirulina powder, and adopt microwave and ultrasonic treatment to assist immersion to obtain a spirulina aqueous solution;
[0052] The microwave and ultrasonic treatment-assisted soaking method is as follows: firstly, microwave treatment is performed for 65 s at a microwave power of 60W and a microwave temperature of 6° C., and then ultrasonic treatment is performed for 7 minutes at an ultrasonic power of 120W and an ultrasonic temperature of 18° C. According to the above The microwave and ultrasonic treatments were alternately performed three times; the soaking parameters were controlled as follows: the soaking pressure was 0.09MPa, the soaking temperature was 8°C, and the soaking time was 3h;
[0053] (3) Enzymatic hydrolysis: under dark and argon or nitrogen protection conditions, add mixed enzyme A to the above-mentioned Spirulina aqueous solution for enzymatic hydrolysis, and use microwave treatment to assist enzymatic hydrolysis, then add mixed enzyme B, and further use ultrasonic treatment to assist enzyme solution to obtain an enzymatic hydrolysis mixture;
[0054] Described microwave treatment: microwave power is 95W, microwave time is 120s, microwave temperature is 7 ℃; Described ultrasonic treatment: ultrasonic power is 120W, ultrasonic wave time is 9min, ultrasonic temperature is 25 ℃; Described mixed enzyme A is protease, Cellulase, pectinase, hemicellulase, and peroxidase are in a mass ratio = 3:2:1:0.3:0.2; before adding mixed enzyme B, the temperature is raised to 50°C and an enzyme inhibitor is added to adjust the pH. Inactivate the enzyme, cool down to 10°C and then add mixed enzyme B; after ultrasonic treatment to assist enzymolysis, heat the material to 50°C and add an enzyme inhibitor to adjust the pH to inactivate the enzyme, and then cool down to 1°C to obtain an enzymatic hydrolysis mixture; Mixed enzyme B is lignase, cellulase, amylase, esterase, lipoxygenase, and pectinase according to the mass ratio=2:2:1:0.5:0.5:1.5;
[0055] (4) Filtration: under dark and argon or nitrogen protection conditions, use an ultrafiltration membrane to filter the above-mentioned enzymatic hydrolysis mixture, remove the filtration mother liquor, and obtain a chlorophyll semi-finished product;
[0056] (5) drying: under dark and argon or nitrogen protection conditions, the chlorophyll semi-finished product obtained above is sent into a dryer for drying, sealed, packaged, and protected from light to obtain the chlorophyll; the decompression drying The temperature is 30 DEG C, the relative air humidity is less than or equal to 65%, the drying time is 30min, and the chlorophyll water content is less than 20%.

Example Embodiment

[0057] Example 3
[0058] A process for extracting chlorophyll from spirulina, comprising the following steps:
[0059] (1) Raw material pretreatment: Wash the fresh spirulina, treat it in the dark at 2°C for 82min, then pulverize it at 7°C until it passes through a 60-mesh sieve, put it into a centrifuge tube equipped with steel balls, and pour it into liquid nitrogen. When the liquid nitrogen has just evaporated, cover the centrifuge tube cover and place it on the vortexer for vortexing, and vortex to the spirulina powder that has passed through a 270-mesh sieve; during the vortexing process, the centrifuge tube is repeatedly inverted 5 times;
[0060] (2) immersion: under dark and argon or nitrogen protection conditions, add deionized water immersion treatment to the above-mentioned spirulina powder, and adopt microwave and ultrasonic treatment to assist immersion to obtain a spirulina aqueous solution;
[0061] The microwave and ultrasonic treatment-assisted soaking methods are as follows: firstly, microwave treatment is performed for 50 s at a microwave power of 52W and a microwave temperature of 3° C., and then ultrasonic treatment is performed for 4 minutes under an ultrasonic power of 110W and an ultrasonic temperature of 19° C. according to the above. The microwave and ultrasonic treatments were alternately performed twice; the soaking parameters were controlled as follows: the soaking pressure was 0.06MPa, the soaking temperature was 2°C, and the soaking time was 2.6h;
[0062] (3) Enzymatic hydrolysis: under dark and argon or nitrogen protection conditions, add mixed enzyme A to the above-mentioned Spirulina aqueous solution for enzymatic hydrolysis, and use microwave treatment to assist enzymatic hydrolysis, then add mixed enzyme B, and further use ultrasonic treatment to assist enzyme solution to obtain an enzymatic hydrolysis mixture;
[0063] Described microwave treatment: microwave power is 87W, microwave time is 80s, microwave temperature is 3 ℃; Described ultrasonic treatment: ultrasonic power is 110W, ultrasonic wave time is 4min, ultrasonic temperature is 19 ℃; Described mixed enzyme A is protease, Cellulase, pectinase, hemicellulase, and peroxidase were in a mass ratio = 3:1.2:1:0.3:0.12; before adding mixed enzyme B, the temperature was raised to 42 °C and an enzyme inhibitor was added to adjust pH Inactivate the enzyme, cool down to 2°C, and then add mixed enzyme B; after the assisted enzymolysis by ultrasonic treatment, the material is heated to 43°C, and an enzyme inhibitor is added to adjust the pH to inactivate the enzyme, and then the temperature is lowered to 3°C to obtain an enzymatic hydrolysis mixture; Mixed enzyme B is lignase, cellulase, amylase, esterase, lipoxygenase, and pectinase according to the mass ratio=1.2:2:1:0.5:0.5:1.2;
[0064] (4) Filtration: under dark and argon or nitrogen protection conditions, use an ultrafiltration membrane to filter the above-mentioned enzymatic hydrolysis mixture, remove the filtration mother liquor, and obtain a chlorophyll semi-finished product;
[0065] (5) drying: under dark and argon or nitrogen protection conditions, the chlorophyll semi-finished product obtained above is sent into a dryer for drying, sealed, packaged, and protected from light to obtain the chlorophyll; the decompression drying The temperature is 27°C, the relative air humidity is less than or equal to 65%, the drying time is 25min, and the drying is performed until the chlorophyll water content is lower than 20%.
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