CN109134414B - Method for separating purple sweet potato anthocyanin by using glutaraldehyde modified magnetic chitosan nanoparticles - Google Patents
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- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
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Abstract
A method for separating purple sweet potato anthocyanin by glutaraldehyde modified magnetic chitosan nanoparticles comprises the steps of preparing purple sweet potato juice rich in anthocyanin by using purple sweet potatoes as a raw material, performing adsorption separation on anthocyanin in the purple sweet potato juice by using the glutaraldehyde modified magnetic chitosan nanoparticles as an adsorption separation material, and finally performing elution, vacuum concentration and freeze drying to obtain a purple sweet potato anthocyanin product. The method for separating purple sweet potato anthocyanin by using glutaraldehyde modified magnetic chitosan nanoparticles, disclosed by the invention, has the advantages of simple process, easiness in operation, convenience in popularization and application, low energy consumption in the separation process, no harmful waste discharge and high comprehensive utilization rate of resources. According to the method for separating the purple sweet potato anthocyanin by using the glutaraldehyde modified magnetic chitosan nanoparticles, the prepared purple sweet potato anthocyanin is high in purity, and can be widely applied to health products and food industries as an antioxidant, a natural pigment, a nutrition enhancer and the like.
Description
Technical Field
The invention belongs to the technical field of food processing, relates to a preparation method of purple sweet potato anthocyanin, and particularly relates to a method for separating purple sweet potato anthocyanin by using glutaraldehyde modified magnetic chitosan nanoparticles.
Background
Anthocyanins (anthocyanidins) are a flavonoid compound widely found in nature, and have attracted attention in recent years because anthocyanins in purple potatoes are stable as a water-soluble natural pigment. It has good anti-aging effect as natural pigment; can promote blood circulation; the eyesight can be enhanced, and the fatigue of eyes can be properly relieved; has certain promoting and protecting effects on cardiopulmonary function of human body, and can also prevent senile dementia, etc.
At present, the direct diet is mainly adopted for human bodies to take anthocyanin in daily life, and the cost for separating and extracting the anthocyanin is relatively high, the steps are complicated, and how to extract the anthocyanin with high efficiency and low cost becomes a research focus. After the surface of the magnetic nano-particles is modified by the chitosan, the magnetic nano-particles not only have the characteristics of large surface area and good magnetism, but also effectively prevent agglomeration after modification, and show good biocompatibility. According to the invention, by adopting the characteristics of large surface area of magnetic chitosan and easiness in separation, recovery and reutilization, purple sweet potato juice rich in anthocyanin is prepared by taking purple sweet potatoes as raw materials, then glutaraldehyde modified magnetic chitosan nanoparticles are taken as adsorption separation materials to carry out adsorption separation on anthocyanin in the purple sweet potato juice, and finally the purple sweet potato anthocyanin product is obtained through elution, vacuum concentration and freeze-drying. The method has the characteristics of simple process, easy operation, energy conservation, no harmful waste discharge, high comprehensive utilization rate of resources, high product purity, convenient popularization and application and the like. The product prepared by the method can be widely applied to health products and food industries as an antioxidant, a natural pigment, a nutrition enhancer and the like.
Disclosure of Invention
The invention aims to provide a method for separating purple sweet potato anthocyanin by using glutaraldehyde modified magnetic chitosan nanoparticles, which is used for adsorbing and separating anthocyanin in purple sweet potato juice by using the glutaraldehyde modified magnetic chitosan nanoparticles as an adsorption and separation material according to the characteristics of large surface area and easiness in separation and recovery of magnetic chitosan.
In order to achieve the purpose, the technical scheme of the invention is as follows:
a method for separating purple sweet potato anthocyanin by glutaraldehyde modified magnetic chitosan nanoparticles comprises the following steps:
s1: respectively preparing purple sweet potato juice with the pH value of 3-5 and glutaraldehyde modified magnetic chitosan nanoparticles;
s2: placing glutaraldehyde modified magnetic chitosan nanoparticles into purple sweet potato juice with the pH value of 3-5, and performing saturated adsorption under the shaking condition;
s3: separating glutaraldehyde modified magnetic chitosan nanoparticles which are saturated and adsorb purple sweet potato anthocyanin under the condition of an external magnetic field;
s4: placing the separated glutaraldehyde modified magnetic chitosan nanoparticles saturated and adsorbing the purple sweet potato anthocyanin in an organic solvent containing hydrochloric acid and at the temperature of 55-65 ℃ for ultrasonic treatment to obtain a purple sweet potato anthocyanin eluent;
s5: and (4) carrying out vacuum concentration and freeze drying on the purple sweet potato anthocyanin eluent to obtain the purple sweet potato anthocyanin freeze-dried powder.
As a preferable technical scheme of the invention, the preparation of the purple sweet potato juice with the pH of 3-5 in the step S1 comprises the following steps:
a1) cleaning and crushing fresh purple sweet potatoes, adding water and high-temperature resistant α -amylase into the crushed purple sweet potatoes, and performing enzymolysis for 1.5-2 hours to obtain purple sweet potato pulp;
a2) squeezing the purple sweet potato pulp to obtain purple sweet potato juice;
a3) adjusting the pH value of the purple sweet potato juice, adding saccharifying enzyme, and carrying out saccharification and enzymolysis at the enzymolysis temperature of 50-55 ℃;
a4) adjusting the pH value of the purple sweet potato juice subjected to saccharification and enzymolysis, adding pectinase, and carrying out enzymolysis at the enzymolysis temperature of 30;
a5) sequentially adding bentonite and active carbon, stirring, standing, filtering, and blending with hydrochloric acid to obtain purple sweet potato juice with pH of 3-5;
as a preferred technical solution of the present invention, the preparing of the glutaraldehyde-modified magnetic chitosan nanoparticle in step S1 includes the steps of:
b1) FeCl is added3·6H2O and FeCl2·4H2Adding O into water protected by nitrogen, stirring, adjusting the pH to 10-11, separating the product under the condition of an external magnetic field after water bath at 80 ℃, and fully washing the product to be neutral by using distilled water to obtain Fe3O4A magnetic core solution;
b2) preparing acetic acid solution containing chitosan, and mixing Fe3O4Mixing the magnetic core solution, the acetic acid solution, cyclohexane and n-hexanol to obtain emulsion, slowly adding Triton X-100 into the obtained emulsion until the emulsion is changed from black to white, continuously adding Triton X-100 until the emulsion is gradually changed into black, adding NaOH, carrying out water bath, and respectively washing with absolute ethyl alcohol and water to obtain a magnetic chitosan nanoparticle solution;
b3) the obtained magnetic chitosan nanoparticle solution is fixed in Fe3O4And (3) oscillating and activating in glutaraldehyde, and washing with water to obtain the glutaraldehyde modified magnetic chitosan nanoparticles.
As a preferred technical solution of the present invention, the step b1) specifically includes: adding FeCl into nitrogen-protected water3·6H2O and FeCl2·4H2O, and F3+With Fe2+Stirring for 5-10 minutes at a molar ratio of 2:1 to allow FeCl3·6H2O and FeCl2·4H2Fully dissolving O in water, dropwise adding concentrated ammonia water into the stable water solution until the pH value is 10-11, then carrying out water bath at 80 ℃ for 30min, separating the obtained product under the condition of an external magnetic field, and washing the product with distilled water to be neutral to obtain Fe3O4Magnetic core solution.
As a preferable embodiment of the present invention, said Fe3O4The volume ratio of the magnetic core solution to the acetic acid solution to the cyclohexane to the n-hexanol is 4:4:11: 6.
As a preferable technical scheme of the invention, the step b3) specifically comprises the step of containing the obtained magnetic chitosan nano-particles in 10mg/m L Fe3O4And (3) oscillating and activating the concentration for 60min at the rotating speed of 150rpm in a glutaraldehyde environment of 0.5 mol/L, and then washing for 2-3 times to obtain the glutaraldehyde modified magnetic chitosan nanoparticles.
As a preferred technical solution of the present invention, the step S2 specifically includes: placing the glutaraldehyde modified magnetic chitosan nano-particles into purple sweet potato juice with the pH value of 3-5, and oscillating at 150rpm for 1 hour to complete saturated adsorption.
As a preferable technical scheme, the step S4 specifically comprises the steps of placing the separated glutaraldehyde modified magnetic chitosan nanoparticles with saturated adsorption of purple sweet potato anthocyanin in a methanol solution or ethanol solution containing hydrochloric acid at 55-65 ℃, wherein the concentration of the hydrochloric acid is 0.1 mol/L, and treating the mixture for 30-45 min under an ultrasonic environment with power of 150-250W to obtain the purple sweet potato anthocyanin eluent.
As a preferable technical scheme of the invention, the methanol solution or the ethanol solution is 85-95% by volume of methanol solution or 65-75% by volume of ethanol solution.
As a preferred technical solution of the present invention, the step S5 specifically includes: pumping the purple sweet potato anthocyanin eluent into a vacuum concentration device, and carrying out vacuum concentration under the conditions that the vacuum degree is-0.6 to-0.9 Mpa and the temperature is 75-85 ℃ until no methanol or ethanol exists in a concentrated solution; collecting a purple sweet potato anthocyanin concentrated solution and an evaporation solution, pre-freezing the collected purple sweet potato anthocyanin concentrated solution for 5-6 hours at-18 ℃, and then freeze-drying for 24-30 hours under the conditions that the temperature is-50 ℃ and the vacuum degree is 25-50 Pa to obtain the purple sweet potato anthocyanin freeze-dried powder with the purity of 95-98%.
Compared with the prior art, the invention has the beneficial effects that: the method for separating purple sweet potato anthocyanin by using glutaraldehyde modified magnetic chitosan nanoparticles is simple in process, easy to operate and convenient to popularize and apply; the separation process has low energy consumption, no harmful waste discharge and high comprehensive utilization rate of resources; according to the method for separating the purple sweet potato anthocyanin by using the glutaraldehyde modified magnetic chitosan nanoparticles, the prepared purple sweet potato anthocyanin is high in purity, and can be widely applied to health products and food industries as an antioxidant, a natural pigment, a nutrition enhancer and the like.
The specific implementation mode is as follows:
example 1
1. Preparing concentrated purple sweet potato juice:
fresh purple sweet potatoes are used as raw materials, and are washed, crushed, pulped, liquefied, juiced, secondarily juiced, saccharified, clarified and canned for later use. The specific operation steps are as follows:
(1) crushing the washed purple sweet potatoes, adding water with the mass of 1-2 times of that of the purple sweet potatoes and the temperature of 80-95 ℃ into the crushed purple sweet potatoes, and simultaneously performing treatment according to the weight ratio of 3-5 × 104Adding the addition amount of U/kg into the high temperature resistantα -amylase at the temperature, and carrying out enzymolysis for 1.5-2 hours at the temperature of 95 ℃;
(2) feeding the purple sweet potato pulp subjected to enzymolysis into a juice extractor for primary squeezing, adding water at 45-50 ℃ into the squeezed potato pulp according to the proportion of water to residue of 2:1 for heavy suspension leaching for 30min, carrying out secondary squeezing, combining the purple sweet potato juice squeezed twice, and filtering;
(3) adjusting the pH value of the purple sweet potato juice to 5.0 by using citric acid, and adding 4-4.5 × 105Saccharifying and hydrolyzing U/kg of saccharifying enzyme at 50-55 deg.C for 2 hr;
(4) adjusting the pH value of the saccharified purple sweet potato juice to 4.0 by using citric acid, and adding 1.5-2 × 105Carrying out enzymolysis on U/kg pectinase for 1 hour under the condition that the enzymolysis temperature is 30 ℃;
(5) after enzymolysis, adding bentonite in an amount of 500-800 mg/L, then adding 300-400 mg/L of activated carbon, stirring, standing, filtering, collecting filtrate, and blending with hydrochloric acid to obtain the purple sweet potato juice with pH of 3-5.
2. Preparation of glutaraldehyde modified magnetic chitosan nanoparticles
(1) Preparation of Fe3O4Magnetic core
Adding a certain amount of FeCl into water protected by nitrogen3·6H2O and FeCl2·4H2O(F3+With Fe2+The molar ratio is 2:1), mechanically stirring for 5-10 minutes to fully dissolve the Fe-Fe3O4The magnetic core solution is ready for use.
(2) Preparation of magnetic Chitosan nanoparticles
Preparing a 2% acetic acid solution with the concentration of 1% chitosan by adopting a Triton X-100 reversed-phase water-in-oil microemulsion system of a two-step method, mixing a magnetic core solution, the acetic acid solution, cyclohexane and n-hexanol according to the volume ratio of 4:4:11:6 at the rotation speed of 1800rpm, then slowly adding Triton X-100 until the emulsion is changed from black to white, and gradually changing to transparent black, finally adding 5 mol/L NaOH with the total volume of about 4%, and after carrying out water bath for 120min at 60 ℃, washing with absolute ethyl alcohol and water for 2-3 times respectively to obtain a magnetic chitosan nanoparticle solution for later use.
(3) Preparation of glutaraldehyde modified magnetic chitosan nanoparticles
The obtained magnetic chitosan nano-particles are fixed to 10mg/m L Fe3O4And (3) oscillating and activating the concentration in a glutaraldehyde environment of 0.5 mol/L at the rotating speed of 150rpm for 60min, and washing for 2-3 times to obtain the glutaraldehyde-modified magnetic chitosan nanoparticles for later use.
3. Adsorbing purple sweet potato anthocyanin
Placing a proper amount of magnetic chitosan nanoparticles activated by glutaraldehyde into purple sweet potato juice with the pH value of 3-5, completing saturated adsorption separation under the shaking condition of 150rpm for 1 hour, and after the saturated adsorption is finished, separating the magnetic chitosan nanoparticles subjected to saturated adsorption of purple sweet potato anthocyanin under the condition of an external magnetic field for later use.
4. Eluting purple sweet potato anthocyanin
Placing the magnetic chitosan nanoparticles saturated and adsorbing the purple sweet potato anthocyanin in a 90% methanol solution at 65 ℃, wherein the methanol solution contains 0.1 mol/L hydrochloric acid, treating for 30min under an ultrasonic environment with the power of 200W, and then finishing the elution of the purple sweet potato anthocyanin on the magnetic chitosan nanoparticles, collecting the eluent of the purple sweet potato anthocyanin after the magnetic chitosan nanoparticles are separated by an external magnetic field for later use, wherein the elution rate of the eluent reaches 80.5%.
5. Preparation of purple sweet potato anthocyanin freeze-dried powder
Pumping the purple sweet potato anthocyanin eluent into a vacuum concentration device, completing vacuum concentration at the vacuum degree of-0.6 to-0.9 Mpa and the temperature of 75 to 85 ℃ until no methanol or ethanol exists in the concentrated solution, and collecting the concentrated solution and the evaporated solution of the purple sweet potato anthocyanin. And then pre-freezing the collected purple sweet potato anthocyanin concentrated solution for 5-6 hours at-18 ℃, and freeze-drying for 24-30 hours at-50 ℃ and under the vacuum degree of 25-50 Pa to obtain the purple sweet potato anthocyanin freeze-dried powder with the purity of 96.8%.
Example 2
The difference between the embodiment and embodiment 1 lies in elution of purple sweet potato anthocyanin and preparation of purple sweet potato anthocyanin freeze-dried powder, which are specifically as follows:
4. eluting purple sweet potato anthocyanin
Placing the magnetic chitosan nanoparticles saturated and adsorbing the purple sweet potato anthocyanin in a 70% methanol solution at 55 ℃, treating the methanol solution containing 0.1 mol/L hydrochloric acid for 45min under an ultrasonic environment with the power of 200W, and then completing the elution of the purple sweet potato anthocyanin on the magnetic chitosan nanoparticles, collecting the eluent of the purple sweet potato anthocyanin after the magnetic chitosan nanoparticles are separated by an external magnetic field for later use, wherein the elution rate reaches 67.5%.
5. Preparation of purple sweet potato anthocyanin freeze-dried powder
Pumping the purple sweet potato anthocyanin eluent into a vacuum concentration device, completing vacuum concentration at the vacuum degree of-0.6 to-0.9 Mpa and the temperature of 75 to 85 ℃ until no methanol or ethanol exists in the concentrated solution, and collecting the concentrated solution and the evaporated solution of the purple sweet potato anthocyanin. Pre-freezing the collected purple sweet potato anthocyanin concentrated solution for 5-6 hours at-18 ℃, and freeze-drying for 24-30 hours at-50 ℃ and under the vacuum degree of 25-50 Pa to obtain the purple sweet potato anthocyanin freeze-dried powder with the purity of 95.7%.
Claims (4)
1. A method for separating purple sweet potato anthocyanin by using glutaraldehyde modified magnetic chitosan nanoparticles is characterized by comprising the following steps:
s1: respectively preparing purple sweet potato juice with the pH value of 3-5 and glutaraldehyde modified magnetic chitosan nanoparticles; the purple sweet potato juice with the pH value of 3-5 is prepared by the following steps:
a1) cleaning and crushing fresh purple sweet potatoes, adding water and high-temperature resistant α -amylase into the crushed purple sweet potatoes, and performing enzymolysis for 1.5-2 hours to obtain purple sweet potato pulp;
a2) squeezing the purple sweet potato pulp to obtain purple sweet potato juice;
a3) adjusting the pH value of the purple sweet potato juice, adding saccharifying enzyme, and carrying out saccharification and enzymolysis at an enzymolysis temperature of 50-55 ℃;
a4) adjusting the pH value of the purple sweet potato juice subjected to saccharification and enzymolysis, adding pectinase, and carrying out enzymolysis at the enzymolysis temperature of 30 ℃;
a5) after enzymolysis is finished, sequentially adding bentonite and activated carbon, stirring, standing, filtering, and blending by using hydrochloric acid to obtain purple sweet potato juice with the pH of 3-5;
the preparation method of the glutaraldehyde modified magnetic chitosan nanoparticle comprises the following steps:
b1) adding FeCl into nitrogen-protected water3·6H2O and FeCl2·4H2O, and Fe3+With Fe2+Stirring for 5-10 minutes at a molar ratio of 2:1 to allow FeCl3·6H2O and FeCl2·4H2Fully dissolving O in water, dropwise adding concentrated ammonia water into the stable water solution until the pH value is 10-11, then carrying out water bath at 80 ℃ for 30min, separating the obtained product under the condition of an external magnetic field, and washing the product with distilled water to be neutral to obtain Fe3O4A magnetic core solution;
b2) preparing acetic acid solution containing chitosan, and mixing Fe3O4Mixing the magnetic core solution, the acetic acid solution, cyclohexane and n-hexanol according to a volume ratio of 4:4:11:6 to obtain emulsion, slowly adding Triton X-100 into the obtained emulsion until the emulsion is changed from black to white, continuously adding Triton X-100 until the emulsion is gradually changed into black, adding NaOH, carrying out water bath, and respectively washing with absolute ethyl alcohol and water to obtain a magnetic chitosan nanoparticle solution;
b3) the obtained magnetic chitosan nano-particles are fixed to 10mg/m L Fe3O4Performing oscillation activation at the rotating speed of 150rpm for 60min in the environment of 0.5 mol/L glutaraldehyde according to the concentration, and then washing for 2-3 times to obtain glutaraldehyde modified magnetic chitosan nanoparticles;
s2: placing glutaraldehyde modified magnetic chitosan nanoparticles into purple sweet potato juice with the pH value of 3-5, and performing saturated adsorption under the shaking condition;
s3: separating glutaraldehyde modified magnetic chitosan nanoparticles which are saturated and adsorb purple sweet potato anthocyanin under the condition of an external magnetic field;
s4, placing the separated glutaraldehyde modified magnetic chitosan nanoparticles saturated and adsorbing purple sweet potato anthocyanin into a methanol solution or ethanol solution containing hydrochloric acid at the temperature of 55-65 ℃, wherein the concentration of the hydrochloric acid is 0.1 mol/L, and treating for 30-45 min under an ultrasonic environment with the power of 150-250W to obtain purple sweet potato anthocyanin eluent;
s5: and (4) carrying out vacuum concentration and freeze drying on the purple sweet potato anthocyanin eluent to obtain the purple sweet potato anthocyanin freeze-dried powder.
2. The method for separating purple sweet potato anthocyanin by using glutaraldehyde-modified magnetic chitosan nanoparticles as claimed in claim 1, wherein the step S2 specifically comprises: placing the glutaraldehyde modified magnetic chitosan nano-particles into purple sweet potato juice with the pH value of 3-5, and oscillating at 150rpm for 1 hour to complete saturated adsorption.
3. The method for separating purple sweet potato anthocyanin from glutaraldehyde-modified magnetic chitosan nanoparticles as claimed in claim 1, wherein the methanol solution or ethanol solution is 85-95% by volume or 65-75% by volume.
4. The method for separating purple sweet potato anthocyanin by using glutaraldehyde-modified magnetic chitosan nanoparticles as claimed in claim 1, wherein the step S5 specifically comprises: pumping the purple sweet potato anthocyanin eluent into a vacuum concentration device, and carrying out vacuum concentration under the conditions that the vacuum degree is-0.6 to-0.9 Mpa and the temperature is 75-85 ℃ until no methanol or ethanol exists in a concentrated solution; collecting a purple sweet potato anthocyanin concentrated solution and an evaporation solution, pre-freezing the collected purple sweet potato anthocyanin concentrated solution for 5-6 hours at-18 ℃, and then freeze-drying for 24-30 hours under the conditions that the temperature is-50 ℃ and the vacuum degree is 25-50 Pa to obtain the purple sweet potato anthocyanin freeze-dried powder with the purity of 95-98%.
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| CN102687881A (en) * | 2012-05-02 | 2012-09-26 | 烟台大学 | Preparation method for pure natural green high-anthocyanin purple sweet potato beverage |
| CN106046079A (en) * | 2016-05-27 | 2016-10-26 | 合肥工业大学 | Method for rapidly separating and preparing peonidin-3-caffeoyl sophoroside-5-glucoside from purple sweet potatoes |
| US20170182188A1 (en) * | 2015-12-29 | 2017-06-29 | King Abdulaziz University | Method for preparing chitosan-coated magnetic nanoparticles for protein immobilization |
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| CN102687881A (en) * | 2012-05-02 | 2012-09-26 | 烟台大学 | Preparation method for pure natural green high-anthocyanin purple sweet potato beverage |
| US20170182188A1 (en) * | 2015-12-29 | 2017-06-29 | King Abdulaziz University | Method for preparing chitosan-coated magnetic nanoparticles for protein immobilization |
| CN106046079A (en) * | 2016-05-27 | 2016-10-26 | 合肥工业大学 | Method for rapidly separating and preparing peonidin-3-caffeoyl sophoroside-5-glucoside from purple sweet potatoes |
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