CN112048025A - Zinc supplement water-soluble soybean polysaccharide zinc chelate as well as preparation method and application thereof - Google Patents
Zinc supplement water-soluble soybean polysaccharide zinc chelate as well as preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a water-soluble soybean polysaccharide zinc chelate of a zinc supplement agent, and a preparation method and application thereof. The method comprises the following steps: dissolving soybean polysaccharide in deionized water, adding a certain amount of zinc chloride solution, adjusting the pH value of a reaction system by using an alkaline solution or an acidic solution, then carrying out constant-temperature oscillation reaction, adding 3-4 times of volume of absolute ethyl alcohol into a reaction solution after the reaction is cooled, standing for 8-12 h at 0-4 ℃, centrifuging to remove a supernatant, and washing precipitates by using 95% volume fraction of ethyl alcohol, acetone and diethyl ether respectively. And (3) after washing, collecting the precipitate, and freeze-drying the obtained precipitate to obtain the water-soluble soybean polysaccharide zinc chelate. The water-soluble soybean polysaccharide zinc chelate provided by the invention is simple in synthesis method, high in zinc content of the chelate and higher in oxidation resistance.
Description
Technical Field
The invention relates to the field of synthesis of novel health-care products, in particular to a water-soluble soybean polysaccharide zinc chelate of a zinc supplement agent, and a preparation method and application thereof.
Background
The water-soluble soybean polysaccharide is a water-soluble polysaccharide substance extracted from soybean of Leguminosae, has many excellent functional properties such as emulsifying property, oxidation resistance, antibacterial property, film forming property, protein particle stabilizing effect under acidic condition, anti-caking property and foam stability, and can be used as emulsifier, dispersant, stabilizer, adsorbent, etc.
Zinc is one of important trace elements necessary for human bodies and animal bodies, and the content of the trace elements contained in the human bodies is second to that of iron. Zinc is a component of many enzymes in human bodies or an activator of the enzymes, and zinc deficiency in the human bodies causes malnutrition and cardiovascular diseases, and can cause various cancers in severe cases. Most of common zinc supplement agents in the market are inorganic zinc, are not easy to absorb, and have certain toxicity after being taken too much.
Compared with the polysaccharide zinc chelate synthesized by other methods, the zinc content is lower, the average content is about 6mg/g (the preparation of the rice germ zinc polysaccharide and the antioxidant activity thereof [ J ] in food industry science and technology, 2017,038(014): 238-.
The metal chelate with oxidation resistance can prevent or delay food oxidation, improve the stability of the food, prolong the storage period and bring better food safety to consumers. Therefore, the development of a novel healthy, nontoxic and antioxidant organic zinc supplement agent, namely water-soluble soybean polysaccharide zinc, has important significance.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention aims to provide a water-soluble soybean polysaccharide zinc chelate which can be used as a zinc supplement preparation and has oxidation resistance.
The water-soluble soybean polysaccharide zinc chelate provided by the invention is a novel biological zinc supplement agent, and can solve the problems of inorganic zinc in the aspects of human body absorption, toxicity, emission pollution and the like.
Meanwhile, the novel water-soluble soybean polysaccharide zinc chelate of the zinc supplement agent can also improve the oxidation resistance.
In order to achieve the above object, a first aspect of the present invention provides a method for preparing a water-soluble soybean polysaccharide zinc chelate as a zinc supplement.
The second aspect of the invention is to provide the water-soluble soybean polysaccharide zinc chelate of the zinc supplement agent prepared by the preparation method.
The third aspect of the present invention is to provide the above-mentioned water-soluble soybean polysaccharide as a zinc supplement or an antioxidant.
In order to achieve the above object of the present invention, the following technical solutions are adopted.
The invention provides a preparation method of a water-soluble soybean polysaccharide zinc chelate of a zinc supplement agent, which comprises the following steps:
dissolving water-soluble soybean polysaccharide powder in deionized water, adding a zinc chloride solution, wherein the zinc chloride solution is prepared by dissolving zinc chloride in hydrochloric acid, adding an alkaline solution or an acidic solution to adjust the pH value of a reaction system, and carrying out constant-temperature oscillation reaction for a period of time to form a reaction solution containing a polysaccharide-zinc chelate;
and carrying out alcohol precipitation and centrifugation on the reaction solution to obtain a precipitate, namely a crude product, and washing, drying and grinding the crude product to obtain the water-soluble soybean polysaccharide zinc chelate.
Further, the constant temperature condition is soaking in a constant temperature water bath.
Further, in the preparation method, the mass ratio of the water-soluble soybean polysaccharide to the zinc chloride is 12: 1-1: 1;
preferably, the mass ratio of the water-soluble soybean polysaccharide to the zinc chloride is 10: 1-6: 1; more preferably at a mass ratio of 8: 1.
Further, in the preparation method, the pH value of a reaction system of the water-soluble soybean polysaccharide and the zinc chloride is 3-7;
preferably, the pH of the reaction system of the water-soluble soybean polysaccharide and the zinc chloride is 4-6; more preferably at a pH of 5.
Further, in the preparation method, the reaction temperature of the water-soluble soybean polysaccharide and zinc chloride is 30-70 ℃;
preferably, the reaction temperature of the water-soluble soybean polysaccharide and zinc chloride is 40-60 ℃; more preferably, the reaction temperature is 50 ℃.
Further, in the preparation method, the reaction time of the water-soluble soybean polysaccharide and the zinc chloride is 0.5-2.5 hours;
preferably, the reaction time of the water-soluble soybean polysaccharide and the zinc chloride is 1-2 h; more preferably the reaction time is 1.5 h.
Further, the alcohol precipitation is to add 3-4 times of volume of absolute ethyl alcohol into the reaction solution and then place the reaction solution at 0-4 ℃ for standing for 8-12 hours; preferably, the alcohol precipitation condition is that 4 times of volume of absolute ethyl alcohol is added and then the mixture is placed at 4 ℃ and stands for 12 hours.
Further, the washing is to wash the precipitate with an organic solvent, wherein the organic solvent is ethanol with the volume fraction of 95%, acetone and diethyl ether; the drying is freeze drying.
The water-soluble soybean polysaccharide zinc chelate prepared by any one of the preparation methods.
The water-soluble soybean polysaccharide zinc chelate is applied to the preparation of zinc supplementing or antioxidant medicaments.
The invention also provides the application of the water-soluble soybean polysaccharide zinc chelate in a biological zinc supplement agent;
meanwhile, the invention also provides application of the water-soluble soybean polysaccharide zinc chelate as an antioxidant.
Compared with the prior art, the invention has the beneficial effects that:
(1) the preparation method is simple to operate, the source of the raw material water-soluble soybean polysaccharide is rich, the water-soluble soybean polysaccharide zinc chelate is obtained through reaction, and the condition selection is optimized, so that the zinc content can be greatly improved.
(2) The water-soluble soybean polysaccharide zinc chelate can be used as a zinc supplement agent, can provide safe, nontoxic and easily absorbed zinc elements for organisms, can also be used as an antioxidant in food, and has good oxidation resistance.
(3) The content of the water-soluble soybean polysaccharide zinc chelate prepared by the method is 20-30 mg/g.
Drawings
FIG. 1 is a graph of a zinc standard curve;
FIG. 2 is a graph of DPPH clearance for test example 1;
FIG. 3 is a graph showing OH clearance in test example 1.
Detailed Description
The following description of the embodiments of the present invention is provided in connection with the accompanying drawings and examples, but the invention is not limited thereto. It is noted that the processes described below, if not specifically described in detail, are all realizable or understandable by those skilled in the art with reference to the prior art. The reagents or apparatus used are not indicated to the manufacturer, and are considered to be conventional products available by commercial purchase.
The method for measuring the zinc content in the water-soluble soybean polysaccharide zinc chelate prepared in each example comprises the following steps:
determination of zinc standard curve: precisely sucking 1mL of 1000mg/L zinc standard solution into a 100mL volumetric flask, diluting the zinc standard solution to a scale with 5% nitric acid to obtain standard intermediate solution with the concentration of 10mg/L, respectively and precisely sucking 0mL, 1.00mL, 2.00mL, 4.00mL, 8.00mL and 10.00mL of the zinc standard intermediate solution into the 100mL volumetric flask, adding 5% nitric acid solution to a constant volume to a scale, and uniformly mixing. Measuring absorbance on an atomic spectrophotometer; the zinc standard curve can be plotted (see fig. 1).
The zinc content determination method comprises the following specific steps: accurately weighing 0.2g of solid sample into a microwave digestion tank, adding 4mL of nitric acid, and carrying out microwave digestion. And cooling, taking out the digestion tank, and removing acid to about 1mL on an electric hot plate at 140-160 ℃. And (3) after the digestion tank is cooled, transferring the digestion solution into a 250mL volumetric flask, washing the digestion tank with a small amount of water for 2-3 times, combining the washing solutions in the volumetric flask, fixing the volume to the scale with water, and uniformly mixing for later use. And measuring a light absorption value on an atomic spectrophotometer, and calculating the zinc content.
The content of zinc in the water-soluble soybean polysaccharide zinc chelate is calculated according to the following formula:
X＝[(ρ-ρ0)V]/m
in the formula:
x-the content of zinc in the sample, and the unit is mg/kg;
rho-mass concentration of zinc in the sample solution, and the unit is mg/L;
ρ0-mass concentration of zinc in the blank solution in mg/L;
v-constant volume of the sample digestive juice, unit is mL;
m-sample weighing or transfer volume, unit is g.
Example 1
Adding water-soluble soybean polysaccharide into a beaker, adding a certain amount of deionized water, stirring to completely dissolve the polysaccharide, and adding a zinc chloride solution (the zinc chloride solution is prepared by dissolving zinc chloride in 1% hydrochloric acid) after the soybean polysaccharide is completely dissolved, wherein the mass ratio of the water-soluble soybean polysaccharide to the zinc chloride is 8: 1. After mixing uniformly, slowly adding an acidic solution or an alkaline solution to adjust the pH value of the reaction solution to 5, then continuously reacting in a water bath at 50 ℃ for 1.5h, after the reaction is cooled, adding four times of volume of absolute ethyl alcohol, standing at 4 ℃ for 12h, centrifuging at 8000r/min for 5min to remove supernatant, and washing precipitates with 95% ethanol, acetone and diethyl ether respectively. And collecting the precipitate. And freeze-drying the precipitate to obtain the water-soluble soybean polysaccharide zinc chelate.
The absorbance of the water-soluble soybean polysaccharide zinc chelate is measured by an atomic spectrophotometer, the mass concentration of zinc in the water-soluble soybean polysaccharide zinc chelate is 0.87mg/L according to a standard curve of zinc by the atomic spectrophotometer, and the mass concentration of the diluted zinc in the water-soluble soybean polysaccharide zinc chelate prepared by the embodiment is 24.03mg/g by multiplying the mass concentration by a dilution factor after calculation according to a formula.
Example 2
Adding water-soluble soybean polysaccharide into a beaker, adding a certain amount of deionized water, stirring to completely dissolve the polysaccharide, and adding a zinc chloride solution (the zinc chloride solution is prepared by dissolving zinc chloride in hydrochloric acid) after the soybean polysaccharide is completely dissolved, wherein the mass ratio of the water-soluble soybean polysaccharide to the zinc chloride is 10: 1. After mixing uniformly, slowly adding an acidic solution or an alkaline solution to adjust the pH value of the reaction solution to 6, then continuing to react in a water bath at 30 ℃ for 0.5h, after the reaction is cooled, adding anhydrous ethanol with three times of volume, standing at 4 ℃ for 8h, centrifuging at 8000r/min for 5min to remove supernatant, and washing precipitates with ethanol with the volume fraction of 95%, acetone and ether respectively. And collecting the precipitate. And freeze-drying the precipitate to obtain the water-soluble soybean polysaccharide zinc chelate.
The absorbance of the water-soluble soybean polysaccharide zinc chelate is measured by an atomic spectrophotometer, the mass concentration of zinc in the water-soluble soybean polysaccharide zinc chelate is 0.62mg/L according to a standard curve of zinc by the atomic spectrophotometer, and the mass concentration of the diluted zinc in the water-soluble soybean polysaccharide zinc chelate prepared by the embodiment is 18.43mg/g by multiplying the mass concentration by a dilution factor after calculation according to a formula.
Example 3
Adding water-soluble soybean polysaccharide into a beaker, adding a certain amount of deionized water, stirring to completely dissolve the polysaccharide, and adding a zinc chloride solution (the zinc chloride solution is prepared by dissolving zinc chloride in hydrochloric acid) after the soybean polysaccharide is completely dissolved, wherein the mass ratio of the water-soluble soybean polysaccharide to the zinc chloride is 12: 1. After uniform mixing, slowly adding an acidic solution or an alkaline solution to adjust the pH value of the reaction solution to 3, then continuously reacting in a water bath at 60 ℃ for 2.5h, after the reaction is cooled, adding four times of volume of absolute ethyl alcohol, standing at 4 ℃ for 8h, centrifuging at 8000r/min for 5min to remove supernatant, and washing precipitates with 95% ethanol, acetone and diethyl ether respectively. And collecting the precipitate. And freeze-drying the precipitate to obtain the water-soluble soybean polysaccharide zinc chelate.
The absorbance of the water-soluble soybean polysaccharide zinc chelate is measured by an atomic spectrophotometer, the mass concentration of zinc in the water-soluble soybean polysaccharide zinc chelate is 0.59mg/L according to a standard curve of zinc by the atomic spectrophotometer, and the mass concentration of the diluted zinc in the water-soluble soybean polysaccharide zinc chelate prepared by the embodiment is 19.05mg/g by multiplying the mass concentration by a dilution factor after calculation according to a formula.
Example 4
Adding water-soluble soybean polysaccharide into a beaker, adding a certain amount of deionized water, stirring to completely dissolve the polysaccharide, and adding a zinc chloride solution (the zinc chloride solution is prepared by dissolving zinc chloride in hydrochloric acid) after the soybean polysaccharide is completely dissolved, wherein the mass ratio of the water-soluble soybean polysaccharide to the zinc chloride is 1: 1. After mixing uniformly, slowly adding an acidic solution or an alkaline solution to adjust the pH value of the reaction solution to 4, then continuing to react in a water bath at 70 ℃ for 2h, after the reaction is cooled, adding anhydrous ethanol with three times of volume, standing at 0 ℃ for 8h, centrifuging at 8000r/min for 5min to remove supernatant, and washing the precipitate with ethanol with the volume fraction of 95%, acetone and ether respectively. And collecting the precipitate. And freeze-drying the precipitate to obtain the water-soluble soybean polysaccharide zinc chelate.
The absorbance of the water-soluble soybean polysaccharide zinc chelate is measured by an atomic spectrophotometer, the mass concentration of zinc in the water-soluble soybean polysaccharide zinc chelate is 0.50mg/L according to a standard curve of zinc by the atomic spectrophotometer, and the mass concentration of the diluted zinc in the water-soluble soybean polysaccharide zinc chelate prepared by the embodiment is 16.08mg/g by multiplying the mass concentration by a dilution factor after calculation according to a formula.
Example 5
Adding water-soluble soybean polysaccharide into a beaker, adding a certain amount of deionized water, stirring to completely dissolve the polysaccharide, and adding a zinc chloride solution (the zinc chloride solution is prepared by dissolving zinc chloride in hydrochloric acid) after the soybean polysaccharide is completely dissolved, wherein the mass ratio of the water-soluble soybean polysaccharide to the zinc chloride is 6: 1. After uniform mixing, slowly adding an acidic solution or an alkaline solution to adjust the pH value of the reaction solution to 7, then continuously reacting in a water bath at 40 ℃ for 1.5h, after the reaction is cooled, adding four times of volume of absolute ethyl alcohol, standing at 4 ℃ for 12h, centrifuging at 8000r/min for 5min to remove supernatant, and washing precipitates with 95% ethanol, acetone and diethyl ether respectively. And collecting the precipitate. And freeze-drying the precipitate to obtain the water-soluble soybean polysaccharide zinc chelate.
The absorbance of the water-soluble soybean polysaccharide zinc chelate is measured by an atomic spectrophotometer, the mass concentration of zinc in the water-soluble soybean polysaccharide zinc chelate is 0.74mg/L according to a standard curve of zinc by the atomic spectrophotometer, and the mass concentration of the diluted zinc in the water-soluble soybean polysaccharide zinc chelate prepared by the embodiment is 21.24mg/g by multiplying the mass concentration by a dilution factor after calculation according to a formula.
Test example 1
The water-soluble soybean polysaccharide zinc prepared in example 1 was tested for oxidation resistance by the following specific test method:
the experimental method comprises the following steps:
(1) the cleaning effect of water-soluble soybean polysaccharide zinc on DPPH is tested: preparing water-soluble soybean polysaccharide zinc into water solutions with different gradient concentrations, adding 0.2mmol/L DPPH absolute ethyl alcohol solution 2mL into 1mL of water-soluble soybean polysaccharide zinc solution, mixing, standing in the dark for 30min, and measuring absorbance A at 517nm1. Clearance (%) - (1- (A)1-A2)/A0]X 100, equation A0(distilled water instead of sample) as the absorbance of control; a. the1Is the absorbance of the sample; a. the2(distilled water instead of DPPH) is the background absorbance of the sample. The Vc and water-soluble soybean polysaccharides were tested for DPPH clearance in the same manner.
(2) The water-soluble soybean polysaccharide zinc has the effect of eliminating OH free radicals: preparing water-soluble soybean polysaccharide zinc into water solutions with different gradient concentrations, and sequentially adding 1.0mL of 9mmol/L ferrous sulfate solution, 1.0mL of 9mmol/L salicylic acid solution and 1.0mL of 9mmol/L hydrogen peroxide solution into the polysaccharide zinc solution. Shaking and mixing the mixed solution uniformly, reacting for 15min in water bath at 37 ℃, and measuring the absorbance A at 510nm1. Clearance (%) - (A)1-A0)/A0]X 100, equation A0(distilled water instead of sample) as the absorbance of control; a. the1Is the absorbance of the sample. The Vc and water-soluble soybean polysaccharide were tested for OH scavenging effect in the same manner.
The concentration of the water-soluble soybean polysaccharide zinc solution in the above experiments (1) and (2) was divided into 5 gradient concentrations of 1mg/mL, 2mg/mL, 3mg/mL, 4mg/mL and 5mg/mL, respectively.
As shown in fig. 2 and fig. 3, in the in vitro antioxidant experiment, although the antioxidant property of the water-soluble soybean polysaccharide zinc is lower than that of Vc, compared with the water-soluble soybean polysaccharide, the water-soluble soybean polysaccharide zinc chelate has better antioxidant effect and more advantages. The clearance of the water-soluble soybean polysaccharide zinc chelate to DPPH is increased along with the increase of the concentration of the water-soluble soybean polysaccharide zinc, and the maximum clearance is 63.31 percent; the clearance rate of OH free radicals is increased along with the increase of the concentration of the water-soluble soybean polysaccharide zinc, and the maximum clearance rate is 64.29%. In conclusion, the water-soluble soybean polysaccharide zinc DPPH and OH free radicals have a good scavenging effect; further shows that the water-soluble soybean polysaccharide zinc has good oxidation resistance.
The above examples are only preferred embodiments of the present invention, which are intended to be illustrative and not limiting, and those skilled in the art should understand that they can make various changes, substitutions and alterations without departing from the spirit and scope of the invention.
Claims (10)
1. A preparation method of a water-soluble soybean polysaccharide zinc chelate of a zinc supplement agent is characterized by comprising the following steps:
(1) preparation of water-soluble soybean polysaccharide zinc chelate
Dissolving water-soluble soybean polysaccharide powder in deionized water, adding a zinc chloride solution, wherein the zinc chloride solution is prepared by dissolving zinc chloride in hydrochloric acid, adjusting the pH value with an acidic solution or an alkaline solution, and performing oscillation reaction at constant temperature to form a reaction solution containing a polysaccharide zinc chelate;
(2) precipitating with ethanol, washing and drying
And (2) carrying out alcohol precipitation and centrifugation on the reaction liquid obtained in the step (1), obtaining a precipitate which is a crude product, and washing, drying and grinding the crude product to obtain the water-soluble soybean polysaccharide zinc chelate.
2. The preparation method according to claim 1, wherein the mass ratio of the water-soluble soybean polysaccharide to the zinc chloride in the step (1) is 12: 1-1: 1, and the pH of a reaction system of the water-soluble soybean polysaccharide and the zinc chloride is 3-7.
3. The preparation method according to claim 2, wherein the mass ratio of the water-soluble soybean polysaccharide to the zinc chloride in the step (1) is 10: 1-6: 1, and the pH of a reaction system of the water-soluble soybean polysaccharide and the zinc chloride is 4-6.
4. The method according to claim 1, wherein the constant temperature condition in step (1) is soaking in a constant temperature water bath.
5. The preparation method according to claim 1, wherein the reaction temperature of the water-soluble soybean polysaccharide and the zinc chloride in the step (1) is 30-70 ℃, and the reaction time of the water-soluble soybean polysaccharide and the zinc chloride in the step (1) is 0.5-2.5 h.
6. The preparation method according to claim 5, wherein the reaction temperature of the water-soluble soybean polysaccharide and the zinc chloride in the step (1) is 40-60 ℃, and the reaction time of the water-soluble soybean polysaccharide and the zinc chloride in the step (1) is 1-2 h.
7. The preparation method according to claim 1, wherein the alcohol precipitation in the step (2) is that the reaction solution is added with 3-4 times volume of absolute ethyl alcohol and then placed at 0-4 ℃ for standing for 8-12 hours; washing the precipitate with an organic solvent, wherein the organic solvent is ethanol with the volume fraction of 95%, acetone and diethyl ether; the drying is freeze drying.
8. The preparation method according to claim 7, wherein the alcohol precipitation in the step (2) is carried out by adding 4 times of volume of absolute ethanol into the reaction solution and standing at 4 ℃ for 12 hours.
9. The water-soluble soybean polysaccharide zinc chelate prepared by the preparation method of any one of claims 1 to 8.
10. The use of the water-soluble soybean polysaccharide zinc chelate of claim 9 in the preparation of a zinc-supplementing or antioxidant medicament.
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