Spaceflight purple corncob anthocyanin as well as extraction method and application thereof

Abstract

The invention belongs to the field of anthocyanin extraction, and particularly relates to aerospace purple corncob anthocyanin and an extraction method and application thereof. The extraction method comprises the following steps: crushing spaceflight purple corn, mixing the crushed spaceflight purple corn with water, and then adding compound bacteria for fermentation; after fermentation is completed, suction filtration is conducted, filter residues obtained through suction filtration are dissolved in an organic solvent, oscillation treatment and ultrasonic treatment are conducted in sequence, then concentration suction filtration is conducted, freeze-drying sublimation and grinding are conducted on filtrate, and the anthocyanin is obtained. The extraction method disclosed by the invention has the advantages of high anthocyanin extraction rate, high yield and high preparation efficiency, the extracted anthocyanin has relatively strong immunomodulatory effect and antioxidant effect, and in addition, the extraction method is simple, low in cost and green and environment-friendly.

Description

Technical Field

[0001] The invention belongs to the field of anthocyanin extraction, and specifically relates to a space purple corncob anthocyanin and an extraction method and application thereof. Background Art

[0002] Anthocyanins are a class of flavonoid polyphenol compounds found in plants. They are a class of water-soluble natural pigments widely found in plants in nature. They are the main coloring substances in fruits, vegetables, and flowers that are obtained by hydrolysis of anthocyanins. Anthocyanins have antioxidant, vision-enhancing, and immune-enhancing effects.

[0003] Currently known methods for extracting anthocyanins include solvent extraction, enzyme extraction, ultrasonic assisted extraction, etc. Specifically:

[0004] 1. Solvent extraction method: Anthocyanin is a water-soluble pigment and has better stability in an acidic environment, so the extraction of anthocyanin often uses an acidic aqueous solution or an acidic organic solution extraction method.

[0005] 2. Enzymatic extraction method: The enzymatic method mainly targets the composition of plant cell walls, using the high efficiency and specificity of cellulase, pectinase, α-amylase, etc. to degrade or hydrolyze the components of the cell walls, destroy the structure of the cell walls, and expose the effective ingredients to the maximum extent, dissolve them in the solvent, thereby increasing the extraction rate of anthocyanins.

[0006] 3. Ultrasonic-assisted extraction method: Ultrasonic waves will close in the liquid medium and generate an instantaneous pressure, causing the plant cells to rupture. The effective ingredients in the cells are accelerated to melt out, and the ultrasonic vibration is homogenized, which can make the sample extraction uniform.

[0007] However, the above method has disadvantages such as low preparation efficiency, low output and high cost. Summary of the invention

[0008] Based on the above technical background, the main purpose of the present invention is to provide an aerospace purple corncob anthocyanin and an extraction method and application thereof to overcome the deficiencies in the prior art.

[0009] In order to achieve the above-mentioned invention object, the technical solution adopted by the present invention includes:

[0010] The first aspect of the present invention is to provide a method for extracting anthocyanins from space purple corn cobs, the extraction method comprising the following steps:

[0011] Step 1, crush the aerospace purple corn, mix it in water, and then add composite bacteria for fermentation;

[0012] Step 2: After the fermentation is completed, the filtrate is filtered, and the filtrate obtained by the filtration is dissolved in an organic solvent, and then subjected to shaking treatment and ultrasonic treatment in sequence, and then concentrated and filtered, and the filtrate is freeze-dried, sublimated, and ground to obtain anthocyanins.

[0013] Preferably, in step 1,

[0014] The spaceflight purple corn is selected from one or more of purple corn kernels, purple corn leaves, purple corn cobs, purple corn stalks, and purple corn silks;

[0015] The space purple corn is crushed into powder, and the powder is mixed in water according to the material-liquid ratio of the space purple corn powder to water of 1 g: (40-70) mL, and 20-120 U / g of cellulase is added, and the mixture is treated at 45-55° C. for 1-3 hours.

[0016] Preferably, in step 1,

[0017] After adding cellulase for treatment, 1×10 5 The composite bacteria is fermented at 20-42°C for 10-30h;

[0018] The composite bacteria is a mixture of Aspergillus niger, lactic acid bacteria, yeast, cellulase and pectinase in a mass ratio of 1-15:1-15:0.5-15:0.5-15:0.5-15.

[0019] Preferably, in step 2,

[0020] The filter residue obtained by suction filtration is dissolved in 30% to 70% ethanol.

[0021] Preferably, in step 2,

[0022] The shaking treatment conditions are: constant temperature shaking treatment at 30-60°C for 2-6 hours.

[0023] Preferably, in step 2,

[0024] The ultrasonic treatment conditions are: ultrasonic treatment at 100W-400W for 20-80min and ultrasonic temperature of 30-50°C.

[0025] The second aspect of the present invention is to provide a space purple corncob anthocyanin prepared according to the preparation method described in the first aspect of the present invention.

[0026] The third aspect of the present invention is to provide an application of the aerospace purple corncob anthocyanins described in the second aspect of the present invention in the fields of anti-oxidation and immune enhancement.

[0027] The beneficial effects of the present invention are:

[0028] (1) The extraction method of the present invention uses aerospace purple corn cobs as the extraction raw material, has the advantages of low cost, turning waste into treasure, being green and environmentally friendly, and increasing the added value of aerospace purple corn cobs. At the same time, a microbial fermentation extraction method and a solvent extraction method are combined to extract anthocyanins, so that the extraction method has the advantages of high anthocyanin extraction rate, high yield, and high preparation efficiency;

[0029] (2) The anthocyanins extracted by the extraction method of the present invention have the effect of enhancing immune regulation and have a strong antioxidant effect, and have good application prospects in the field of enhancing immune regulation and antioxidant effects;

[0030] (3) The extraction method of the present invention is simple, and no toxic or harmful substances are used or produced during the extraction process. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 A comparison chart of anthocyanins extracted from Example 1, Comparative Example 1 and Comparative Example 2 is shown;

[0032] Figure 2 The results of superoxide dismutase (SOD) assay in the control group, model group, simvastatin group, and anthocyanin group are shown;

[0033] Figure 3 The results of measuring transforming growth factor (TGF-β1) in the control group, model group, simvastatin group and anthocyanin group are shown. DETAILED DESCRIPTION

[0034] The present invention will be described in detail below, and the characteristics and advantages of the present invention will become clearer and more distinct with the description.

[0035] Purple corn cobs in space purple corn are mostly used for incineration, but they have a high content of anthocyanins. The purpose of the present invention is to provide a method for extracting anthocyanins from space purple corn cobs using a lactic acid bacteria-ultrasound-assisted extraction method, wherein the anthocyanins extracted play an important role in anti-oxidation and immune enhancement.

[0036] The first aspect of the present invention is to provide a method for extracting anthocyanins from spaceflight purple corn, the extraction method comprising the following steps:

[0037] Step 1, crush the aerospace purple corn, mix it in water, and then add composite bacteria for fermentation;

[0038] Step 2: After the fermentation is completed, the filtrate is filtered, and the filtrate obtained by the filtration is dissolved in an organic solvent, and then subjected to shaking treatment and ultrasonic treatment in sequence, and then concentrated and filtered, and the filtrate is freeze-dried, sublimated, and ground to obtain anthocyanins.

[0039] The above steps are described in detail below.

[0040] In step 1, the spaceflight purple corn is selected from one or more of purple corn kernels, purple corn leaves, purple corn cobs, purple corn stalks, and purple corn silks; preferably, the purple corn cobs.

[0041] The aerospace purple corn was crushed into powder, and the aerospace purple corn powder was dissolved in water at a material-liquid ratio of 1 g: (40-70) mL. 20-120 U / g of cellulase was added, and the mixture was treated at 45-55 °C for 1-3 h. Then, 1×10 5 The composite bacteria is fermented at 20-42°C for 10-30 hours.

[0042] Preferably, the aerospace purple corn is crushed into powder, and the powder is mixed in water at a material-liquid ratio of 1 g to 50 mL, 80 U / g of cellulase is added, and the mixture is treated at 50° C. for 2 h, and then 1×10 5 The composite bacteria was fermented at 25-30°C for 20 hours.

[0043] The composite bacteria is a mixture of Aspergillus niger, lactic acid bacteria, yeast, cellulase and pectinase in a mass ratio of 1-15:1-15:0.5-15:0.5-15:0.5-15.

[0044] Preferably, the composite bacteria is a mixture of Aspergillus niger, lactic acid bacteria, yeast, cellulase and pectinase in a mass ratio of 1:1:1:1:1.

[0045] In step 2, the filter residue obtained by filtration is dissolved in 30% to 70% ethanol, and then subjected to constant temperature shaking treatment at 30 to 60° C. for 2 to 6 hours, followed by ultrasonic treatment under 100W-400W ultrasonic conditions for 20 to 80 minutes, and the ultrasonic temperature is 30 to 50° C.

[0046] Preferably, the filter residue obtained by filtration is dissolved in 50% ethanol, and then subjected to constant temperature shaking treatment at 50°C for 4 hours, followed by ultrasonic treatment at 300W for 50 minutes, and the ultrasonic temperature is 40°C.

[0047] The second aspect of the present invention is to provide a space purple corncob anthocyanin prepared according to the preparation method described in the first aspect of the present invention.

[0048] The third aspect of the present invention is to provide an application of the aerospace purple corncob anthocyanins described in the second aspect of the present invention in the fields of anti-oxidation and immune enhancement.

[0049] Example

[0050] The present invention is further described below by means of specific examples. These embodiments are only intended to illustrate the present invention but are not intended to limit the scope of the present invention.

[0051] Example 1

[0052] The aerospace purple corn cobs were crushed into powder and dissolved in distilled water with a solid-liquid ratio of 1:50 (g / ml). Then 80 U / g of cellulase was added and treated at 50 °C for 2 h. Then 1×10 5 The composite bacteria is fermented at 27°C for 20 hours. The composite bacteria is a mixture of Aspergillus niger, lactic acid bacteria, yeast, cellulase, and pectinase in a mass ratio of 1:1:1:1:1, and then filtered. The filter residue after filtration is taken and dissolved in 50% ethanol by volume, and then treated at a constant temperature of 50°C for 4 hours, and then treated for 50 minutes under 300W ultrasonic assistance, and the ultrasonic temperature is 40°C. After cooling, it is concentrated and filtered, and the filtrate is freeze-dried and sublimated, and ground to obtain anthocyanins. The extracted anthocyanins contain 0.85±0.05 mg of anthocyanins per 100 grams, which is comparable to the anthocyanins extracted in Comparative Examples 1 and 2. Figure 1 As shown, from Figure 1 It can be seen that the anthocyanins obtained by using aerospace purple corn cobs as the extraction raw material are much higher than those extracted by using blueberries and purple sweet potatoes as raw materials. The content of anthocyanins obtained by using aerospace purple corn cobs as the extraction raw material is 6 to 8 times that of blueberries and 8 to 9 times that of purple sweet potatoes, indicating that using aerospace purple corn cobs as the extraction raw material can greatly increase the extraction amount of anthocyanins.

[0053] Example 2

[0054] The aerospace purple corn cobs were crushed into powder and dissolved in distilled water with a solid-liquid ratio of 1:40 (g / ml). Then 80 U / g of cellulase was added and treated at 50 °C for 2 h. Then 1×10 5 The composite bacteria is fermented at 27°C for 20 hours, wherein the composite bacteria is a mixture of Aspergillus niger, lactic acid bacteria, yeast, cellulase and pectinase in a mass ratio of 1:1:1:1:1, and then filtered. The filter residue after filtration is taken, dissolved in 30% ethanol by volume, and then subjected to constant temperature shaking treatment at 30°C for 6 hours, followed by treatment with 100W ultrasound assistance for 80 minutes, with the ultrasound temperature being 30°C. After cooling, the mixture is concentrated and filtered, and the filtrate is freeze-dried and sublimated, and ground and pulverized to obtain anthocyanins.

[0055] Example 3

[0056] The space purple corn cobs were crushed into powder and dissolved in distilled water with a solid-liquid ratio of 1:70 (g / ml). Then 80 U / g of cellulase was added and treated at 50 °C for 2 h. Then 1×10 5The composite bacteria is fermented at 27°C for 20 hours, wherein the composite bacteria is a mixture of Aspergillus niger, lactic acid bacteria, yeast, cellulase and pectinase in a mass ratio of 1:1:1:1:1, and then filtered, and the filter residue after filtration is taken, dissolved in 70% ethanol by volume, and then subjected to constant temperature shaking treatment at 60°C for 2 hours, and then treated with 400W ultrasound assistance for 20 minutes, the ultrasound temperature being 50°C, and then concentrated and filtered after cooling, and the filtrate is freeze-dried and sublimated, and ground and pulverized to obtain anthocyanins.

[0057] Comparative Example

[0058] Comparative Example 1

[0059] Purple sweet potato was crushed into powder and dissolved in distilled water with a solid-liquid ratio of 1:50 (g / ml). Then 80 U / g of cellulase was added and treated at 50 °C for 2 h. Then 1×10 5 The lactic acid bacteria were fermented at 37°C for 20 hours, and then filtered. The residue after filtration was taken and dissolved in 50% ethanol by volume, and then treated at 50°C for 4 hours, and then treated for 50 minutes under 300W ultrasonic assistance, the ultrasonic temperature was 40°C, and after cooling, concentrated and filtered. The filtrate was vacuum freeze-dried and ground to obtain anthocyanins. The anthocyanins extracted in Example 1 and Comparative Example 2 were compared. Figure 1 shown.

[0060] Comparative Example 2

[0061] Blueberries were crushed into powder and dissolved in distilled water at a solid-liquid ratio of 1:50 (g / ml). Then 80 U / g of cellulase was added and treated at 50 °C for 2 h. Then 1×10 5 The lactic acid bacteria were fermented at 37°C for 20 hours, and then filtered. The filtrate was taken and dissolved in 50% ethanol by volume, and then treated at 50°C for 4 hours, and then treated with 300W ultrasound for 50 minutes, the ultrasound temperature was 40°C, and then concentrated and filtered after cooling. The filtrate was vacuum freeze-dried and ground to obtain anthocyanins. The anthocyanins extracted in Example 1 and Comparative Example 1 were compared. Figure 1 shown.

[0062] Experimental example

[0063] Experimental Example 1 Animal Experiment

[0064] 40 SD rats were selected and divided into a control group, a model group, a simvastatin group and anthocyanidin group, with an average of 10 rats in each group. The model was established by feeding with a high-fat diet: the specific operation was that the control group was fed with ordinary feed all the time, and the other three groups were fed with a high-fat diet. After feeding for four weeks, a hyperlipidemia model was formed. Then the anthocyanidin group and the simvastatin group were gavaged for 4 weeks. The anthocyanidin group was gavaged with anthocyanidin prepared in Example 1, and the simvastatin group was gavaged with simvastatin. Then the animals were killed, and the animal livers were taken for the determination of superoxide dismutase (SOD). The test results are shown as follows: Figure 2 As shown, serum was taken for determination of transforming growth factor (TGF-β1), and the test results were as follows Figure 3 shown.

[0065] Figure 2 and Figure 3 Compared with the control group, *P<0.05, **P<0.01; compared with the model group, #P<0.05, #P<0.01.

[0066] from Figure 2 It can be seen that compared with the control group, the SOD level in the liver of the model group rats was significantly reduced (P<0.01). Compared with the model group, the SOD level in the liver of the anthocyanin group was significantly increased (P<0.01). Superoxide dismutase is an antioxidant metal enzyme in the body. It can catalyze the dismutation of superoxide anion free radicals to generate oxygen and hydrogen peroxide, and plays a vital role in the balance of oxidation and antioxidant in the body. The above results show that anthocyanins extracted from space purple corn cobs have antioxidant effects.

[0067] Figure 3 It can be seen that compared with the control group, the serum TGF-β1 of the model group rats was significantly reduced (P<0.01). Compared with the model group, the serum TGF-β1 level of the anthocyanin group was significantly increased (P<0.01). TGF-β1 is a key regulatory factor in maintaining immune response, which helps mucosal healing and protects colon tissue. This shows that anthocyanins extracted from space purple corn cobs have the effect of enhancing immune regulation.

[0068] The present invention has been described in detail above in conjunction with specific implementations and exemplary examples, but these descriptions cannot be understood as limiting the present invention. Those skilled in the art understand that, without departing from the spirit and scope of the present invention, a variety of equivalent substitutions, modifications or improvements may be made to the technical solution of the present invention and its implementation methods, all of which fall within the scope of the present invention. The scope of protection of the present invention shall be subject to the attached claims.

Claims

1. A method for extracting anthocyanins from aerospace purple corn cobs, characterized in that: The extraction method comprises the following steps: Step 1, crush the aerospace purple corn, mix it in water, and then add composite bacteria for fermentation; Step 2: After the fermentation is completed, the filtrate is filtered, and the filtrate obtained by the filtration is dissolved in an organic solvent, and then subjected to shaking treatment and ultrasonic treatment in sequence, and then concentrated and filtered, and the filtrate is freeze-dried, sublimated, and ground to obtain anthocyanins.

2. The method for extracting anthocyanins from aerospace purple corn cobs according to claim 1, characterized in that: In step 1, The spaceflight purple corn is selected from one or more of purple corn kernels, purple corn leaves, purple corn cobs, purple corn stalks, and purple corn silks; The space purple corn is crushed into powder, and the powder is mixed in water according to the material-liquid ratio of the space purple corn powder to water of 1 g: (40-70) mL, and 20-120 U / g of cellulase is added, and the mixture is treated at 45-55° C. for 1-3 hours.

3. The method for extracting anthocyanins from aerospace purple corn cobs according to claim 2, characterized in that: In step 1, After adding cellulase for treatment, 1×10 5 The composite bacteria is fermented at 20-42°C for 10-30h; The composite bacteria is a mixture of Aspergillus niger, lactic acid bacteria, yeast, cellulase and pectinase in a mass ratio of 1-15:1-15:0.5-15:0.5-15:0.5-15.

4. The method for extracting anthocyanins from aerospace purple corn cobs according to claim 1, characterized in that: In step 2, The filter residue obtained by suction filtration is dissolved in 30% to 70% ethanol.

5. The method for extracting anthocyanins from aerospace purple corn cobs according to claim 1, characterized in that: In step 2, The shaking treatment conditions are: constant temperature shaking treatment at 30-60°C for 2-6 hours.

6. The method for extracting anthocyanins from aerospace purple corn cobs according to claim 1, characterized in that: In step 2, The ultrasonic treatment conditions are: ultrasonic treatment at 100W-400W for 20-80min and ultrasonic temperature of 30-50°C.

7. Anthocyanins extracted according to the method for extracting anthocyanins from spaceflight purple corncobs according to any one of claims 1 to 6.

8. Use of the anthocyanin according to claim 7 in the field of anti-oxidation and immune enhancement.

Images