CN107034260B - Preparation method of nano-zinc oyster powder - Google Patents
Preparation method of nano-zinc oyster powder Download PDFInfo
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- CN107034260B CN107034260B CN201710502158.2A CN201710502158A CN107034260B CN 107034260 B CN107034260 B CN 107034260B CN 201710502158 A CN201710502158 A CN 201710502158A CN 107034260 B CN107034260 B CN 107034260B
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- 241000237502 Ostreidae Species 0.000 title claims abstract description 110
- 235000020636 oyster Nutrition 0.000 title claims abstract description 110
- 239000000843 powder Substances 0.000 title claims abstract description 65
- 239000011701 zinc Substances 0.000 title claims abstract description 59
- 229910052725 zinc Inorganic materials 0.000 title claims abstract description 59
- 238000002360 preparation method Methods 0.000 title claims abstract description 33
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims abstract description 36
- 229960001763 zinc sulfate Drugs 0.000 claims abstract description 35
- 229910000368 zinc sulfate Inorganic materials 0.000 claims abstract description 35
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 34
- 239000002002 slurry Substances 0.000 claims abstract description 20
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 claims abstract description 17
- 239000007864 aqueous solution Substances 0.000 claims abstract description 17
- 238000006243 chemical reaction Methods 0.000 claims abstract description 15
- 238000001694 spray drying Methods 0.000 claims abstract description 14
- 239000000243 solution Substances 0.000 claims abstract description 11
- 238000000034 method Methods 0.000 claims abstract description 9
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- 238000004140 cleaning Methods 0.000 claims abstract description 7
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- 239000012535 impurity Substances 0.000 claims abstract description 7
- 238000005360 mashing Methods 0.000 claims abstract description 7
- 238000002156 mixing Methods 0.000 claims abstract description 7
- 238000000926 separation method Methods 0.000 claims abstract description 6
- 230000009849 deactivation Effects 0.000 claims abstract description 3
- 102000004142 Trypsin Human genes 0.000 claims description 9
- 108090000631 Trypsin Proteins 0.000 claims description 9
- 239000012588 trypsin Substances 0.000 claims description 9
- 238000005238 degreasing Methods 0.000 claims description 8
- 238000000605 extraction Methods 0.000 claims description 6
- 239000012065 filter cake Substances 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 6
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- 238000007873 sieving Methods 0.000 claims description 6
- 239000002904 solvent Substances 0.000 claims description 5
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 2
- 238000001291 vacuum drying Methods 0.000 claims description 2
- 239000002105 nanoparticle Substances 0.000 abstract description 34
- 239000002245 particle Substances 0.000 abstract description 14
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- 108090000623 proteins and genes Proteins 0.000 description 3
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- 229940091251 zinc supplement Drugs 0.000 description 3
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
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- 230000002035 prolonged effect Effects 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 description 1
- 238000003917 TEM image Methods 0.000 description 1
- 206010048259 Zinc deficiency Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
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- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000013522 chelant Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000007071 enzymatic hydrolysis Effects 0.000 description 1
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 1
- 238000011067 equilibration Methods 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
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- 239000007788 liquid Substances 0.000 description 1
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- IYDGMDWEHDFVQI-UHFFFAOYSA-N phosphoric acid;trioxotungsten Chemical compound O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.OP(O)(O)=O IYDGMDWEHDFVQI-UHFFFAOYSA-N 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 230000002797 proteolythic effect Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000011669 selenium Substances 0.000 description 1
- 229910052711 selenium Inorganic materials 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 239000011686 zinc sulphate Substances 0.000 description 1
- 235000009529 zinc sulphate Nutrition 0.000 description 1
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
- C07K1/113—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides without change of the primary structure
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Abstract
The invention belongs to the technical field of aquatic product processing, and particularly relates to a preparation method of nano-zinc oyster powder, which comprises the following preparation steps: 1) pretreatment of oysters: removing shell of Concha Ostreae, cleaning to remove impurities, draining, mashing tissue to obtain slurry, and defatting to obtain defatted Concha Ostreae powder; 2) oyster enzymolysis: preparing the defatted oyster powder prepared in the step 1) into an aqueous solution, performing enzymolysis, enzyme deactivation, cooling, performing centrifugal separation, taking supernate, performing vacuum concentration, and performing spray drying to obtain oyster peptide powder; 3) nano zinc of oyster peptide: preparing the oyster peptide powder obtained in the step 2) into an aqueous solution, adding zinc sulfate at room temperature, adding the zinc sulfate to a certain concentration, uniformly mixing, adjusting the pH value to react, and after the reaction is finished, carrying out vacuum concentration and spray drying on the reaction solution to obtain the nano zinc oyster powder. The method can be used for preparing the oyster peptide-zinc nanoparticles with the particle size range of 28-102 nm, and then the nano-zinc oyster powder can be prepared.
Description
Technical Field
The invention belongs to the technical field of aquatic product processing, and particularly relates to a preparation method of nano-zinc oyster powder.
Background
According to the prior literature report, the oyster is the food raw material with the highest zinc content and is a famous zinc supplement food. Because zinc plays a vital role in important physiological processes such as growth and development of human bodies, reproductive inheritance and the like, and zinc deficiency is frequent, researches on developing zinc supplement products by using oysters pay attention to the zinc supplement products. Not many studies on this are currently being made, but all focus on the use of oyster (poly) peptides or proteolytic digests in combination with zinc sulphate to form chelates.
Researches in recent years show that the nano mineral element supplement has the characteristic of high absorption rate. The biological template method for regulating and controlling the nanocrystallization of mineral elements by utilizing biomolecules such as protein peptides, polysaccharides and the like has the advantage of mild reaction conditions and is more and more paid attention by researchers. At present, research shows that calcium, iron and selenium can form nanoparticles under the action of protein peptide or polysaccharide, however, whether protein peptide and zinc can form nanoparticles is not reported, and whether the oyster peptide zinc chelate has a nanostructure is unclear.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a preparation method of nano-zinc oyster powder, and the oyster peptide-zinc nanoparticles with the particle size range of 28-102 nm can be prepared by the method, so that the nano-zinc oyster powder can be prepared.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention provides a preparation method of nano-zinc oyster powder, which comprises the following preparation steps:
1) pretreatment of oysters: removing shell of Concha Ostreae, cleaning to remove impurities, draining, mashing tissue to obtain slurry, and defatting to obtain defatted Concha Ostreae powder;
2) oyster enzymolysis: preparing the defatted oyster powder prepared in the step 1) into an aqueous solution, performing enzymolysis, enzyme deactivation, cooling, performing centrifugal separation, taking supernate, performing vacuum concentration, and performing spray drying to obtain oyster peptide powder;
3) nano zinc of oyster peptide: preparing the oyster peptide powder obtained in the step 2) into an aqueous solution with the mass concentration of 0.3-0.5%, adding zinc sulfate at room temperature until the final mass concentration of the zinc sulfate is 0.5-0.9%, uniformly mixing, adjusting the pH value to 6.0-11.0 for reaction, reacting for 1-2 h, concentrating the reaction liquid in vacuum, and performing spray drying to obtain the nano-zinc oyster powder.
In the preparation method, researches show that the nanoparticles can be prepared by simultaneously adjusting the final mass solution concentration of zinc sulfate and the pH value parameter during reaction. If the final mass concentration of the zinc sulfate is too high or the pH value is too high, the system generates precipitation, and when the final mass concentration of the zinc sulfate is too low or the pH value is too low, the system is basically clear, and the nanoparticles cannot be obtained.
Preferably, in step 1) of the present invention, the degreasing treatment specifically includes: extracting the slurry by using a mixed solvent of n-hexane and absolute ethyl alcohol with a volume ratio of 1: 2-4, repeatedly extracting for 2 times, filtering to remove fat, vacuum drying a filter cake, crushing, and sieving with a 100-150-mesh sieve.
Preferably, in step 1) of the present invention, the amount of the solvent added during the degreasing treatment is 2 times the volume of the slurry. In the invention, when the adding amount of the solvent is 2 times of the volume of the slurry, the degreasing effect is better, the waste of the solvent is avoided as much as possible, if the effect is less than 2 times, the effect is not good, and the repeated times must be increased, so that the waste of the solvent is caused, and the preparation period is also prolonged.
Preferably, in step 1) of the present invention, the extraction conditions during the degreasing process are: the temperature is 45-55 ℃, and the extraction time is 8 h. If the extraction time is less than 8 hours, the best degreasing effect is not achieved, and if the extraction time exceeds 8 hours, the preparation period is prolonged.
Preferably, in step 2) of the present invention, the enzymatic hydrolysis specifically comprises the following steps: and (3) carrying out enzymolysis for 1.5-2.5 h by using trypsin at the temperature of 45-49 ℃ and under the condition that the pH value is 8.0-9.0.
Preferably, in the step 2) of the invention, the adding amount of the trypsin is 2500-3500U/g.
Preferably, in the step 2), the defatted oyster powder prepared in the step 1) is prepared into an aqueous solution with the mass concentration of 2-2.5%.
Preferably, in the step 2), the centrifugal separation is carried out for 15-20 min at the rotating speed of 6000-9000 r/min. Under this condition, a better separation effect is obtained.
Preferably, in the step 3) of the invention, the final mass concentration of zinc sulfate is 0.9%. When the final mass concentration of zinc sulfate in the oyster peptide powder aqueous solution reaches 0.9%, better oyster peptide-zinc nanoparticles are obtained, and the particles are uniformly dispersed.
Preferably, in step 3) of the present invention, the reaction has a pH of 11.0. The reaction is carried out at a pH value to obtain better oyster peptide-zinc nanoparticles which are more uniform.
Compared with the prior art, the invention has the beneficial effects that:
1. the method can be used for preparing the nano zinc oyster peptide powder, and the oyster peptide-zinc nanoparticles are stably and uniformly dispersed in the solution within the range of 28-102 nm through the detection of a particle size analyzer.
2. In the invention, the oyster peptide-zinc nanoparticles can be obtained by simultaneously adjusting the final mass concentration of zinc sulfate and the pH value of the solution, and the method is simple and easy to implement.
Drawings
FIG. 1 is a transmission electron micrograph of a sample prepared in example 3 of the present invention.
Detailed Description
The present invention will be further described with reference to the following examples, but the present invention is not limited to these examples.
Example 1
A preparation method of nano-zinc oyster powder comprises the following preparation steps:
1) pretreatment of oysters: removing shells of oysters, cleaning to remove impurities, draining, mashing tissues to obtain slurry, extracting the slurry at 45 ℃ for 2 times by using a n-hexane-absolute ethyl alcohol mixed solvent with a volume ratio of 1:2 which is 2 times of the volume of the slurry, filtering 6 layers of gauze to remove fat, drying a filter cake in vacuum, crushing, and sieving with a 150-mesh sieve to obtain defatted oyster powder;
2) oyster enzymolysis: preparing the defatted oyster powder prepared in the step 1) into an aqueous solution with the mass concentration of 2%, adding 3500U/g of trypsin for enzymolysis for 1.5h at 45 ℃ and under the condition that the pH value is 8.8, inactivating the enzyme for 10min at 95 ℃ after the enzymolysis is finished, cooling, centrifugally separating for 15min at the rotating speed of 9000r/min, taking supernatant, concentrating in vacuum, and then spray-drying to obtain oyster peptide powder;
3) nano zinc of oyster peptide: preparing the oyster peptide powder obtained in the step 2) into an aqueous solution with the mass concentration of 0.4%, adding zinc sulfate at room temperature until the final mass concentration of the zinc sulfate is 0.5%, uniformly mixing, adjusting the pH value to 6.0 for reaction, and after reacting for 1h, carrying out vacuum concentration and spray drying on the reaction liquid to obtain the nano-zinc oyster powder.
Example 2
A preparation method of nano-zinc oyster powder comprises the following preparation steps:
1) pretreatment of oysters: removing shells of oysters, cleaning to remove impurities, draining, mashing tissues to obtain slurry, extracting the slurry at 50 ℃ for 2 times by using a n-hexane-absolute ethyl alcohol mixed solvent with a volume ratio of 1:3 which is 2 times of the volume of the slurry, filtering 6 layers of gauze to remove fat, drying a filter cake in vacuum, crushing, and sieving with a 150-mesh sieve to obtain defatted oyster powder;
2) oyster enzymolysis: preparing the defatted oyster powder prepared in the step 1) into an aqueous solution with the mass concentration of 2.5%, adding 3000U/g trypsin for enzymolysis for 2.5h at 47 ℃ and under the condition that the pH value is 8.5, inactivating the enzyme for 10min at 95 ℃ after the enzymolysis is finished, cooling, centrifugally separating for 17min at the rotating speed of 7000r/min, taking supernate, concentrating in vacuum, and then spray drying to obtain oyster peptide powder;
3) nano zinc of oyster peptide: preparing the oyster peptide powder obtained in the step 2) into an aqueous solution with the mass concentration of 0.3%, adding zinc sulfate at room temperature until the final mass concentration of the zinc sulfate is 0.6%, uniformly mixing, adjusting the pH value to 6.5 for reaction, reacting for 1.5h, and then carrying out vacuum concentration and spray drying on the reaction liquid to obtain the nano-zinc oyster powder.
Example 3
A preparation method of nano-zinc oyster powder comprises the following preparation steps:
1) pretreatment of oysters: removing shells of oysters, cleaning to remove impurities, draining, mashing tissues to obtain slurry, extracting the slurry at 50 ℃ for 2 times by using a n-hexane-absolute ethyl alcohol mixed solvent with a volume ratio of 1:3 which is 2 times of the volume of the slurry, filtering 6 layers of gauze to remove fat, drying a filter cake in vacuum, crushing, and sieving with a 100-mesh sieve to obtain defatted oyster powder;
2) oyster enzymolysis: preparing the defatted oyster powder prepared in the step 1) into an aqueous solution with the mass concentration of 2%, adding 3000U/g trypsin for enzymolysis for 2h at 49 ℃ and the pH value of 9.0, inactivating the enzyme for 10min at 95 ℃ after the enzymolysis is finished, cooling, centrifugally separating for 20min at the rotating speed of 8000r/min, taking supernatant, concentrating in vacuum, and then spray-drying to obtain oyster peptide powder;
3) nano zinc of oyster peptide: preparing the oyster peptide powder obtained in the step 2) into an aqueous solution with the mass concentration of 0.35%, adding zinc sulfate at room temperature until the final mass concentration of the zinc sulfate is 0.9%, uniformly mixing, adjusting the pH value to 11.0 for reaction, and after reacting for 1h, carrying out vacuum concentration and spray drying on the reaction liquid to obtain the nano-zinc oyster powder.
Example 4
A preparation method of nano-zinc oyster powder comprises the following preparation steps:
1) pretreatment of oysters: removing shells of oysters, cleaning to remove impurities, draining, mashing tissues to obtain slurry, extracting the slurry at 55 ℃ for 2 times by using a n-hexane-absolute ethyl alcohol mixed solvent with a volume ratio of 1:4 which is 2 times of the volume of the slurry, filtering 6 layers of gauze to remove fat, drying a filter cake in vacuum, crushing, and sieving with a 100-mesh sieve to obtain defatted oyster powder;
2) oyster enzymolysis: preparing the defatted oyster powder prepared in the step 1) into an aqueous solution with the mass concentration of 2.5%, adding 2500U/g of trypsin for enzymolysis for 2 hours at 48 ℃ and under the condition that the pH value is 8.0, inactivating the enzyme for 10min at 95 ℃ after the enzymolysis is finished, cooling, centrifugally separating for 18min at the rotating speed of 6000r/min, taking supernatant, concentrating in vacuum, and then spray-drying to obtain oyster peptide powder;
3) nano zinc of oyster peptide: preparing the oyster peptide powder obtained in the step 2) into an aqueous solution with the mass concentration of 0.5%, adding zinc sulfate at room temperature until the final mass concentration of the zinc sulfate is 0.8%, uniformly mixing, adjusting the pH value to 7.0 for reaction, and after reacting for 2 hours, carrying out vacuum concentration and spray drying on the reaction liquid to obtain the nano-zinc oyster powder.
Example 5
A preparation method of nano-zinc oyster powder comprises the step 3), adjusting the pH value to 8.0 for reaction. The other steps and parameters were the same as in example 3.
Example 6
A preparation method of nano-zinc oyster powder comprises the step 3), adjusting the pH value to 9.0, and reacting. The other steps and parameters were the same as in example 3.
Example 7
A preparation method of nano-zinc oyster powder comprises the step 3), adjusting the pH value to 10.0, and reacting. The other steps and parameters were the same as in example 3.
In order to fully illustrate that oyster peptide-zinc nanoparticles can be obtained by adjusting the final mass concentration of zinc sulfate and the pH value of a solution, the applicant performs the following screening experiments:
1. materials and instruments
The materials are fresh oyster shells which are removed: supermarkets with Qianzhou Qianzhi; trypsin: 190kU/g of enzyme activity, Pengbo bioengineering, Inc., Nanning; other reagents used in the experiment are analytically pure or more.
DF-101S heat collection type constant temperature heating magnetic stirrer: zhengzhou great wall science and trade company, Inc.; GL-21M high speed refrigerated centrifuge: xiang Yuan centrifuge instruments Ltd; PHS-3C type precision pH meter: shanghai Lei magnetic instrument factories; UV-2550 type ultraviolet spectrophotometer: shimadzu, Japan; ZEN3600 particle size Analyzer, Malvern, UK; JEM-1200EX transmission electron microscope: japan JEOL Ltd.
2. Experimental methods
2.1 initial determination of nanoparticle formation oyster powder was prepared according to the procedure and parameters of example 3 of the present invention, with the difference that the final mass concentration of zinc sulfate and the solution reaction pH were adjusted as required and the light transmittance was measured at 600 nm. Meanwhile, a zinc solution with the same final mass concentration at a corresponding pH value is used as a blank solution, an oyster peptide solution prepared in example 3 is used as a control solution, the lower light transmittance of the two is used as a reference light transmittance, or one of the two is turbid or precipitated, and the other light transmittance is used as the reference light transmittance. When the sample solution does not generate turbidity or precipitation and the light transmittance is reduced by 20% compared with the standard light transmittance, the formation of the nano-particles is judged.
2.2 particle size analysis determination parameters are set as: light source He-Ne laser lamp, wavelength 633nm, incident angle 173 °, viscosity 1.0031, RI 1.330, 25 ± 0.1 ℃, equilibration time 60s, each sample scanned 3 times and averaged.
And 2.3, adsorbing the sample liquid to be detected on a copper net by electron microscope analysis, naturally airing for 5-10 min, then carrying out negative staining by phosphotungstic acid, naturally airing, and then placing on an 80kV transmission electron microscope for observation.
3. Results of the experiment
3.1 initial determination of oyster peptide-Zinc nanoparticle formation
The oyster peptide solution containing 0.1-0.9% of zinc sulfate is adjusted to pH 3-11 respectively, and the light transmittance of the sample solution is measured, as shown in Table 1.
Table 1 light transmittance (%) of mixed solution of oyster peptide and zinc sulfate
Note: the same upper superscript different letters indicate significant differences (p <0.05), the same below; -means turbidity or precipitation.
As can be seen from table 1, the light transmittance of the mixed system increases with increasing pH, but decreases with increasing final mass concentration of zinc sulfate. Conditions capable of forming nanoparticles are summarized in table 2 according to the judgment conditions for nanoparticle formation.
As can be seen from table 2, oyster peptide cannot form nanoparticles with zinc sulfate at low final mass concentrations (0.4% and below). When the final mass concentration of zinc sulfate reaches 0.5% or more, oyster peptide can form nanoparticles with zinc sulfate, the minimum pH required is 6.0, and the minimum pH required for nanoparticle formation is increased along with the increase of the final mass concentration of zinc sulfate, and the pH range is also increased. When the final mass concentration of zinc sulfate reaches 0.9%, the pH value formed by the nanoparticles can reach 11 at most, and the pH range is increased to 3 units.
3.2 particle size distribution of oyster peptide-Zinc nanoparticles
The "nanoparticles" prepared under the conditions judged in the above study as being likely to produce oyster peptide-zinc nanoparticles were measured for particle size distribution using a particle size analyzer to determine whether nanoparticles were formed, and the results are shown in table 3.
TABLE 3 particle size distribution of oyster peptide-Zinc nanoparticles
As is clear from Table 3, under the above conditions, nanoparticles having a particle diameter of 28 to 102nm were formed and the dispersibility was good (PDI < 0.4). It was found by analysis that the particle size increased with a decrease in pH and an increase in the final zinc mass concentration, which was consistent with the results of measurement of light transmittance.
3.3 Electron microscopy of oyster peptide-Zinc nanoparticles
To further confirm the formation of oyster peptide zinc nanoparticles, "nanoparticles" prepared in example 3 were observed by electron microscopy, and the results are shown in fig. 1. The formation of nanoparticles is clearly observed from fig. 1, and the size of the nanoparticles is substantially consistent with the results of the particle size analyzer. The results of the particle size analyzer and the electron microscopy analysis both confirmed the formation of nanoparticles, and the "nanoparticles" obtained in example 3 were the smallest in particle size in the conditions that nanoparticles could be formed.
Claims (10)
1. The preparation method of the nano-zinc oyster powder is characterized by comprising the following preparation steps:
1) pretreatment of oysters: removing shell of Concha Ostreae, cleaning to remove impurities, draining, mashing tissue to obtain slurry, and defatting to obtain defatted Concha Ostreae powder;
2) oyster enzymolysis: preparing the defatted oyster powder prepared in the step 1) into an aqueous solution, performing enzymolysis, enzyme deactivation, cooling, performing centrifugal separation, taking supernate, performing vacuum concentration, and performing spray drying to obtain oyster peptide powder;
3) nano zinc of oyster peptide: preparing the oyster peptide powder obtained in the step 2) into an aqueous solution with the mass concentration of 0.3-0.5%, adding zinc sulfate at room temperature, uniformly mixing, adjusting the pH value to react for 1-2 h, and then carrying out vacuum concentration and spray drying on the reaction solution to obtain nano zinc oyster powder;
when the final mass concentration of the zinc sulfate is 0.5%, adjusting the pH value to be 6;
when the final mass concentration of zinc sulfate is 0.6%, adjusting the pH value to 6.0-6.5;
when the final mass concentration of zinc sulfate is 0.7%, adjusting the pH value to 6.5-7.0;
when the final mass concentration of zinc sulfate is 0.8%, adjusting the pH value to 7.0-9.0;
when the final mass concentration of the zinc sulfate is 0.9%, the pH value is adjusted to 8.0-11.0.
2. The preparation method of the nano-zinc oyster powder according to claim 1, which is characterized by comprising the following steps: in the step 1), the degreasing treatment specifically comprises the following steps: extracting the slurry by using a mixed solvent of n-hexane and absolute ethyl alcohol with a volume ratio of 1: 2-4, repeatedly extracting for 2 times, filtering to remove fat, vacuum drying a filter cake, crushing, and sieving with a 100-150-mesh sieve.
3. The preparation method of the nano-zinc oyster powder according to claim 2, which is characterized in that: in the step 1), the adding amount of the solvent in the degreasing treatment process is 2 times of the volume of the slurry.
4. The preparation method of the nano-zinc oyster powder according to claim 3, which is characterized by comprising the following steps: in the step 1), the extraction conditions in the degreasing treatment process are as follows: the temperature is 45-55 ℃, and the extraction time is 8 h.
5. The preparation method of the nano-zinc oyster powder according to claim 1, which is characterized by comprising the following steps: in the step 2), the enzymolysis specifically comprises the following steps: and (3) carrying out enzymolysis for 1.5-2.5 h by using trypsin at the temperature of 45-49 ℃ and under the condition that the pH value is 8.0-9.0.
6. The preparation method of the nano-zinc oyster powder according to claim 5, which is characterized by comprising the following steps: in the step 2), the adding amount of the trypsin is 2500-3500U/g.
7. The preparation method of the nano-zinc oyster powder according to claim 1, which is characterized by comprising the following steps: in the step 2), the defatted oyster powder prepared in the step 1) is prepared into an aqueous solution with the mass concentration of 2-2.5%.
8. The preparation method of the nano-zinc oyster powder according to claim 1, which is characterized by comprising the following steps: in the step 2), centrifugal separation is carried out for 15-20 min at the rotating speed of 6000-9000 r/min.
9. The preparation method of the nano-zinc oyster powder according to claim 1, which is characterized by comprising the following steps: in the step 3), the final mass concentration of the zinc sulfate is 0.9%.
10. The preparation method of the nano-zinc oyster powder according to claim 1, which is characterized by comprising the following steps: in the step 3), the pH value of the reaction is 11.0.
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| CN108378261A (en) * | 2018-02-08 | 2018-08-10 | 舟山海研食品科技有限公司 | Oyster protein sports drink and preparation method thereof |
| CN109805388A (en) * | 2019-01-25 | 2019-05-28 | 集美大学 | A kind of fucoxanthol-oyster peptide nanoparticles and its preparation method and application |
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