CN112641085A - Antioxidant emulsion and preparation method thereof - Google Patents

Antioxidant emulsion and preparation method thereof Download PDF

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Publication number
CN112641085A
CN112641085A CN202011521095.3A CN202011521095A CN112641085A CN 112641085 A CN112641085 A CN 112641085A CN 202011521095 A CN202011521095 A CN 202011521095A CN 112641085 A CN112641085 A CN 112641085A
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solution
zymolyte
scallop
emulsion
antioxidant
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刘潇阳
王紫旭
阴法文
周大勇
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Dalian Polytechnic University
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Abstract

The invention provides an antioxidant zymolyte emulsion, which comprises 0.25-0.5% of scallop zymolyte, 2.5-10% of linoleic acid, 39.50-39.75% of buffer solution, 30-37.5% of ethanol with the concentration of 95% and 20% of water. According to the invention, the bay scallop zymolyte and the linoleic acid are mixed to prepare the emulsion, so that the oxidation stability of the linoleic acid is improved, meanwhile, the active ingredients of the polypeptide are introduced, the oil-in-water emulsion state is formed with the oil to a certain degree, the water solubility of the oil is increased, and a better antioxidant effect is further provided, the antioxidant activity of the zymolyte has certain continuity, and the emulsion plays a positive role in the storage of the nutritional oil, so that the combination is easier to absorb by a human body, and the nutrition and the safety of the combination are greatly improved.

Description

Antioxidant emulsion and preparation method thereof
Technical Field
The invention relates to the field of food, in particular to a preparation method of an antioxidant active emulsion taking bay scallop zymolyte dry powder as a raw material.
Background
Unsaturated fatty acids such as grease and the like are easily oxidized and decomposed, so that the nutritional function is lost and food rancidity is caused, and the addition of the antioxidant is a protection for functional grease. At present, synthetic antioxidants such as BHT and the like are widely applied, but researches in recent years show that the synthetic antioxidants cause serious harm to human health due to carcinogenic recessiveness, so that natural antioxidants are more and more preferred for production of food enterprises.
At present, the widely used natural antioxidants comprise tea polyphenol, bamboo leaf antioxidant, bay scallop zymolyte and the like, but most of the antioxidants are water-soluble and hardly have antioxidant effect on grease. Wherein, the bay scallop zymolyte is a mixture containing peptides with different molecular weights obtained by hydrolyzing bay scallop protein by protease. Relevant researches show that bay scallop zymolyte has various biological activities of oxidation resistance, tumor resistance, immunoregulation and the like, and particularly has higher oxidation resistance activity and higher safety in the aspect of oxidation resistance activity, so that research interests of scholars at home and abroad are aroused.
Therefore, it is urgent to develop an antioxidant which can exert an antioxidant effect on oils and fats by using a water-soluble natural antioxidant.
Disclosure of Invention
The invention provides a method for preparing a lipid emulsifier and an antioxidant by utilizing the natural antioxidant property of bay scallop zymolyte, so as to solve the problem that the water-soluble natural antioxidant is difficult to have antioxidant effect on grease due to the non-fat-soluble property of the water-soluble natural antioxidant.
In order to realize the aim, the invention provides an antioxidant zymolyte emulsion which comprises 0.25-0.5% of scallop zymolyte, 2.5-10% of linoleic acid, 39.50-39.75% of buffer solution, 30-37.5% of ethanol with the concentration of 95% and 20% of water.
Preferably, the buffer solution comprises 0.2M sodium dihydrogen phosphate solution and 0.2M disodium hydrogen phosphate solution which are mixed according to the volume ratio of 7:18 to prepare 0.2M compound phosphate solution with the pH value of 7.2-7.4.
A method of preparing the antioxidant emulsion of claim 1 comprising the steps of:
s1, dissolving scallop zymolyte in 0.2M phosphate solution with the pH of 7.2-7.4 according to the mass ratio of 1:80-160 to obtain zymolyte solution;
s2, mixing linoleic acid and ethanol with the concentration of 95% according to the volume ratio of 1:3-15 to obtain a linoleic acid solution;
s3, mixing the zymolyte solution, the linoleic acid solution and water according to the volume ratio of 1:1:0.5, and stirring to obtain the antioxidant emulsion.
Preferably, the preparation method of the scallop zymolyte comprises the following steps:
s1, freezing the scallop, then drying in vacuum, and grinding into powder to obtain scallop dry powder;
s2, adding water according to the feed-liquid ratio of 1:100, and adjusting the pH value to 8.0-9.0;
s3, adding alkaline protease into the scallop dry powder used in the S2 and the alkaline protease according to the mass ratio of 100:6, and stirring at 45-50 ℃ until complete hydrolysis;
s4, adding absolute ethyl alcohol into the hydrolyzed solution in the S3, and cooling the solution at a constant temperature of 4-6 ℃ in a dark place until the solution is completely clear and transparent;
s5, removing precipitates and ethanol from the solution obtained after the alcohol in the S4 is reduced;
s6, freezing and then drying in vacuum to obtain the scallop zymolyte.
Preferably, the volume fraction of ethanol contained in the system after the ethanol is added in the step S4 is 75-80%.
Preferably, the ethanol is removed in step S5 by evaporation concentration at 45-50 deg.C under vacuum of 1-20 Pa.
Preferably, the stirring speed in step S3 is 8000-10000 rpm.
Has the advantages that:
according to the invention, the bay scallop zymolyte and the linoleic acid are mixed to prepare the emulsion, so that the oxidation stability of the linoleic acid is improved, and meanwhile, the active ingredients of the polypeptide are introduced, the bay scallop zymolyte can form an oil-in-water emulsion state with oil to a certain extent as a mixed peptide solution, so that the water solubility of the oil is increased, a better antioxidant effect is further provided, the antioxidant activity of the zymolyte has certain persistence, the active effect on the storage of the nutritional oil is achieved, and the combination is easier to be absorbed by a human body. Compared with the synthetic antioxidant used for antioxidation of the oil in the current market, the nutrition and the safety of the synthetic antioxidant are greatly improved, the zymolyte can keep higher antioxidation activity at lower concentration (0.25 percent), the peroxide value inhibition rate approaches to 30 percent when the oil is stored for the fourth day, and the integral hydrophilicity of the oil is obviously improved after the mixed emulsion is prepared, thereby being more beneficial to the addition and utilization of liquid food, such as beverage and the like.
Drawings
FIG. 1 is a graph showing the POV (peroxide value) inhibition ratio as a function of storage time in examples of the present invention.
Detailed Description
Example 1
Bay scallop zymolyte dry powder of 0.25 percent omega
Linoleic acid 2.5% omega
Compounded with 39.75% omega phosphate solution
95% ethanol 37.5% omega
Water 20% omega
1. Preparing bay scallop zymolyte dry powder:
s1: freezing fresh Argopecten irradians in a refrigerator at-30 deg.C for 24 hr, taking out, vacuum freeze drying (freezing temperature of-50 deg.C and vacuum degree of 1Pa), and pulverizing to obtain dried Argopecten irradians powder;
s2: taking 10g of bay scallop dry powder, adding purified water according to the feed-liquid ratio of 1:100, using 0.2M HCl and 0.2M NaOH, and reading by using a pH meter to adjust the pH of the system to 8;
s3: adding alkaline protease (Novozyme 435, 200000U/g, EC 3.1.1.3, from Candida antarctica) into the solution obtained in S2, wherein the ratio of dried powder of Argopecten irradians: putting the solution into a magnetic stirring water bath kettle, and carrying out enzymolysis at 50 ℃ for 5 hours until the scallop protein is completely hydrolyzed, wherein the alkaline protease is 100:6 (the required enzyme activity is 12000U/g);
s4: taking out the S3 solution after enzymolysis, adding 3L of absolute ethyl alcohol into the solution for alcohol reduction, controlling the volume fraction of the ethanol contained in the system after the absolute ethyl alcohol is added to be 75%, and carrying out alcohol reduction on the solution at a constant temperature of 4 ℃ in a dark place for 24h until the solution is completely clear and transparent;
s5: filtering the solution obtained after the alcohol of S4 is reduced to remove the precipitate, then removing the ethanol by evaporation concentration, controlling the temperature of the evaporation concentration to be 50 ℃ and the vacuum degree to be 0.1M Pa;
s6: and (3) putting the solution obtained by the evaporation and concentration of the S5 into a refrigerator at the temperature of-80 ℃ for freezing for 2h, taking out the solution, performing vacuum freeze drying with the vacuum degree of 1Pa, and drying to obtain the bay scallop zymolyte dry powder.
2. Preparation of an antioxidant emulsion (taking 25mL of antioxidant emulsion as an example)
(1) Preparation of solutions
S1: bay scallop zymolyte solution: 62.5mg of the prepared dried bay scallop zymolyte powder is weighed and mixed with 9.9375mL of 0.2M compound phosphate solution (pH7.2). The compound phosphate solution, i.e., PB solution, was prepared by mixing 28mL of 0.2M sodium dihydrogen phosphate solution and 72mL of 0.2M disodium hydrogen phosphate solution (7:18(v: v)), and the concentration used as a buffer solution in this patent was 0.2M.
S2: linoleic acid solution: linoleic acid was measured at 0.625mL and mixed with 9.375mL of 95% ethanol (v/v) (1: 15).
(2) Preparation of emulsions
The solutions obtained from S1 and S2 were mixed with 5mL of water (volume ratio 1:1:0.5), and stirred at 10000rpm for 1min by using a high-speed homogenizer to obtain an emulsion with antioxidant activity.
Example 2
Bay scallop zymolyte dry powder of 0.5 percent omega
Linoleic acid 5% omega
Compounded with 39.5% omega phosphate solution
95% ethanol 35% omega
Water 20% omega
1. Preparing bay scallop zymolyte dry powder:
s1: freezing fresh Argopecten irradians in a refrigerator at-30 deg.C for 24 hr, taking out, vacuum freeze drying (freezing temperature of-50 deg.C and vacuum degree of 1Pa), and pulverizing to obtain dried Argopecten irradians powder;
s2: taking 10g of dried bay scallop powder, adding purified water according to the feed-liquid ratio of 1:100, using 0.2M HCl and 0.2M NaOH, and adjusting the pH value of the system to 8 by reading of a pH meter;
s3: adding alkaline protease (Novozyme 435, 200000U/g, EC 3.1.1.3, from Candida antarctica) into the solution obtained in S2, dried powder of Argopecten irradians: putting the solution into a magnetic stirring water bath kettle, and carrying out enzymolysis at 50 ℃ for 5 hours until the scallop protein is completely hydrolyzed, wherein the alkaline protease is 100:6 (the required enzyme activity is 12000U/g);
s4: taking out the S3 solution after enzymolysis, adding 3L of absolute ethyl alcohol into the solution for alcohol reduction, controlling the volume fraction of the ethanol contained in the system after the absolute ethyl alcohol is added to be 75%, and carrying out alcohol reduction on the solution at a constant temperature of 4 ℃ in a dark place for 24h until the solution is completely clear and transparent;
s5: filtering the solution obtained after the alcohol of S4 is reduced to remove the precipitate, then removing the ethanol by evaporation concentration, controlling the temperature of the evaporation concentration to be 50 ℃ and the vacuum degree to be 0.15M Pa;
s6: placing the solution obtained after the evaporation and concentration of S5 in a refrigerator at the temperature of minus 80 ℃ for freezing for 2h, taking out the solution, performing vacuum freeze drying with the vacuum degree of 1Pa, and drying to obtain bay scallop zymolyte dry powder;
2. preparation of an antioxidant emulsion (taking 25mL of antioxidant emulsion as an example)
(1) Preparation of solutions
S1: bay scallop zymolyte solution: weighing 125mg of bay scallop zymolyte dry powder obtained in S6, and mixing with 10mL of 0.2M compound phosphate solution (pH7.2)
S2: linoleic acid solution: linoleic acid was measured in 2.5mL and mixed with 7.5mL of 95% ethanol (v/v).
(2) Preparation of emulsions
The solutions obtained from S1 and S2 were mixed with 5mL of water (volume ratio 1:1:0.5), and stirred at 10000rpm for 1min by using a high-speed homogenizer to obtain an emulsion with antioxidant activity.
Example 3
Bay scallop zymolyte dry powder of 0.25 percent omega
Linoleic acid 10% omega
Compounded with 39.75% omega phosphate solution
95% ethanol 30% omega
Water 20% omega
1. Preparing bay scallop zymolyte dry powder:
s1: freezing fresh Argopecten irradians in a refrigerator at-30 deg.C for 24 hr, taking out, vacuum freeze drying (freezing temperature of-50 deg.C and vacuum degree of 1Pa), and pulverizing to obtain dried Argopecten irradians powder;
s2: taking 10g of dried bay scallop powder, adding purified water according to the feed-liquid ratio of 1:100, using 0.2M HCl and 0.2M NaOH, and adjusting the pH value of the system to 8 by reading of a pH meter;
s3: adding alkaline protease (Novozyme 435, 200000U/g, EC 3.1.1.3, from Candida antarctica) into the solution obtained in S2, placing the solution in a magnetic stirring water bath, and performing enzymolysis at 50 deg.C for 5 hr until scallop protein is completely hydrolyzed;
s4: taking out the S3 solution after enzymolysis, adding 3L of ethanol into the solution for alcohol reduction, controlling the volume fraction of the ethanol contained in the system after the ethanol is added to be 75%, and carrying out alcohol reduction on the solution at a constant temperature of 4 ℃ in a dark place for 24 hours until the solution is completely clear and transparent;
s5: filtering the solution obtained after the alcohol of S4 is reduced to remove the precipitate, then removing the ethanol by evaporation concentration, controlling the temperature of the evaporation concentration to be 50 ℃ and the vacuum degree to be 0.2 MPa;
s6: placing the solution obtained after the evaporation and concentration of S5 in a refrigerator at the temperature of minus 80 ℃ for freezing for 2h, taking out the solution, performing vacuum freeze drying with the vacuum degree of 1Pa, and drying to obtain bay scallop zymolyte dry powder;
2. preparation of an antioxidant emulsion (taking 25mL of antioxidant emulsion as an example)
(1) Preparation of solutions
S1: bay scallop zymolyte solution: 125mg of bay scallop zymolyte dry powder obtained in the step S6 is weighed and mixed with 9.875mL of 0.2M compound phosphate solution (pH 7.2).
S2: linoleic acid solution: linoleic acid was measured 2.5mL and mixed with 7.5mL of 95% ethanol (v/v).
(2) Preparation of emulsions
The solutions obtained from S1 and S2 were mixed with 5mL of water (volume ratio 1:1:0.5), and stirred at 10000rpm for 1min by using a high-speed homogenizer to obtain an emulsion with antioxidant activity.
And carrying out a simulated accelerated oxidation experiment on the obtained antioxidant active emulsion.
The emulsions prepared in examples 1 to 3 were placed in a constant temperature forced air drying oven under dark conditions, the temperature of the drying oven was set at 40 ℃, samples were taken on days 0,2 and 4, respectively, and the peroxide value (POV) was measured.
Measurement of POV (peroxide value): 0.1mL of the oxidized emulsion was taken, and 4.7mL of 75% ethanol, 0.1mL of a 30% ammonium persulfate solution, and 0.1mL of 20mM ferrous chloride (dissolved in 3.5% hydrochloric acid) were added thereto. After mixing, the solution was incubated for 3 minutes in the absence of light at 40 ℃. The absorbance of the solution was measured at 500nm using a microplate reader.
As shown in FIG. 1, ABC in the figure are the sample groups of examples 1, 2 and 3 for measured and calculated POV (peroxide value) inhibition, wherein the bay scallop substrate content is 0.25%, the linoleic acid content is 2.5%, 5% and 10%, respectively, and the inhibition is calculated by the following formula:
POV inhibition (%) ((sample group-blank)/blank) х 100
The bay scallop zymolyte still keeps stable antioxidant activity under an emulsion system, the emulsifying effect is better along with the increase of the linoleic acid concentration, the antioxidant effect is also increased simultaneously, and the bay scallop zymolyte has stable antioxidant effect which is mainly shown in the fact that the shelf life is prolonged, the bay scallop zymolyte keeps the antioxidant capacity, and by taking the fourth day of storage as an example, when the linoleic acid content is respectively 2.5 percent, 5 percent and 10 percent, the POV inhibition rate of the zymolyte is shown as 22.91 percent, 28.48 percent and 29.20 percent, and has an ascending trend compared with 4.02 percent, 7.69 percent and 9.24 percent on the 0 th day.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be able to cover the technical solutions and the inventive concepts of the present invention within the technical scope of the present invention.

Claims (7)

1. An antioxidant zymolyte emulsion is characterized by comprising 0.25-0.5% of scallop zymolyte, 2.5-10% of linoleic acid, 39.50-39.75% of buffer solution, 30-37.5% of ethanol with the concentration of 95% and 20% of water.
2. The emulsion of claim 1, wherein the buffer comprises 0.2M sodium dihydrogen phosphate solution and 0.2M disodium hydrogen phosphate solution mixed at a volume ratio of 7:18 to form a 0.2M complex phosphate solution with a pH of 7.2-7.4.
3. A method for preparing the antioxidant emulsion of claim 1, comprising the steps of:
s1, dissolving scallop zymolyte in 0.2M phosphate solution with the pH of 7.2-7.4 according to the mass ratio of 1:80-160 to obtain zymolyte solution;
s2, mixing linoleic acid and ethanol with the concentration of 95% according to the volume ratio of 1:3-15 to obtain a linoleic acid solution;
s3, mixing the zymolyte solution, the linoleic acid solution and water according to the volume ratio of 1:1:0.5, and stirring to obtain the antioxidant emulsion.
4. The method for preparing the antioxidant emulsion as claimed in claim 3, wherein the preparation method of the scallop zymolyte comprises the following steps:
s1, freezing the scallop, then drying in vacuum, and grinding into powder to obtain scallop dry powder;
s2, adding water according to the feed-liquid ratio of 1:100, and adjusting the pH value to 8.0-9.0;
s3, adding alkaline protease into the scallop dry powder used in the S2 and the alkaline protease according to the mass ratio of 100:6, and stirring at 45-50 ℃ until complete hydrolysis;
s4, adding absolute ethyl alcohol into the hydrolyzed solution in the S3, and cooling the solution at a constant temperature of 4-6 ℃ in a dark place until the solution is completely clear and transparent;
s5, removing precipitates and ethanol from the solution obtained after the alcohol in the S4 is reduced;
s6, freezing and then drying in vacuum to obtain the scallop zymolyte.
5. The method for preparing an antioxidant emulsion as claimed in claim 4, wherein the volume fraction of ethanol contained in the system after the ethanol is added in step S4 is 75-80%.
6. The method of claim 4, wherein the ethanol is removed in step S5 by evaporation concentration at 45-50 deg.C under a vacuum of 0.1-0.2M Pa.
7. The method as claimed in claim 3, wherein the stirring speed in step S3 is 8000-10000 rpm.
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Citations (5)

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Publication number Priority date Publication date Assignee Title
JPH09124504A (en) * 1995-10-30 1997-05-13 Morinaga Milk Ind Co Ltd Antimicrobial peptide composition and its production
CN107523601A (en) * 2017-10-11 2017-12-29 山东大学 The preparation method and applications of scallop edge whitening peptide
CN109122906A (en) * 2018-09-14 2019-01-04 南京财经大学 A kind of rice protein lotion and preparation method thereof
CN111748595A (en) * 2019-12-24 2020-10-09 浙江海洋大学 Active polypeptide with antioxidant effect
CN111869747A (en) * 2020-08-03 2020-11-03 东北农业大学 Preparation process of high-linoleic acid antioxidant powdered oil

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH09124504A (en) * 1995-10-30 1997-05-13 Morinaga Milk Ind Co Ltd Antimicrobial peptide composition and its production
CN107523601A (en) * 2017-10-11 2017-12-29 山东大学 The preparation method and applications of scallop edge whitening peptide
CN109122906A (en) * 2018-09-14 2019-01-04 南京财经大学 A kind of rice protein lotion and preparation method thereof
CN111748595A (en) * 2019-12-24 2020-10-09 浙江海洋大学 Active polypeptide with antioxidant effect
CN111869747A (en) * 2020-08-03 2020-11-03 东北农业大学 Preparation process of high-linoleic acid antioxidant powdered oil

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