CN113197299A - Preparation method of oyster powder with microcirculation improving effect - Google Patents
Preparation method of oyster powder with microcirculation improving effect Download PDFInfo
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- CN113197299A CN113197299A CN202110535402.1A CN202110535402A CN113197299A CN 113197299 A CN113197299 A CN 113197299A CN 202110535402 A CN202110535402 A CN 202110535402A CN 113197299 A CN113197299 A CN 113197299A
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- 241000237502 Ostreidae Species 0.000 title claims abstract description 85
- 235000020636 oyster Nutrition 0.000 title claims abstract description 85
- 239000000843 powder Substances 0.000 title claims abstract description 40
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- 238000002360 preparation method Methods 0.000 title claims abstract description 23
- 230000000694 effects Effects 0.000 title claims abstract description 17
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- 241001523579 Ostrea Species 0.000 description 1
- 208000031481 Pathologic Constriction Diseases 0.000 description 1
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 1
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- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
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- 108010007119 flavourzyme Proteins 0.000 description 1
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- 230000007365 immunoregulation Effects 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/06—Enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
- A23L17/50—Molluscs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/27—Removal of unwanted matter, e.g. deodorisation or detoxification by chemical treatment, by adsorption or by absorption
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/28—Removal of unwanted matter, e.g. deodorisation or detoxification using microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/30—Physical treatment, e.g. electrical or magnetic means, wave energy or irradiation
- A23L5/32—Physical treatment, e.g. electrical or magnetic means, wave energy or irradiation using phonon wave energy, e.g. sound or ultrasonic waves
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Life Sciences & Earth Sciences (AREA)
- Polymers & Plastics (AREA)
- Nutrition Science (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Meat, Egg Or Seafood Products (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to the field of oyster product processing, and in particular relates to a preparation method of oyster powder with an effect of improving microcirculation. The preparation method comprises the steps of crushing, degreasing, deodorization, enzymolysis, alcoholysis, drying and the like of the fresh oyster. The oyster is subjected to composite treatment by two steps of enzymolysis and alcoholysis, the reaction end point is detected by detecting the molecular weight of the enzymolysis reaction and the alcoholysis reaction under proper reaction conditions, and the obtained mixture of a plurality of compounds with small molecular weight is obtained and is capped by ethanol, so that the oyster has a better effect on resisting microcirculation disturbance.
Description
Technical Field
The invention relates to the field of oyster product processing, and in particular relates to a preparation method of oyster powder with an effect of improving microcirculation.
Background
Microcirculation is a vascular network between arterioles and venules, plays an important role in regulating the energy metabolism and the functions of tissues and organs of an organism, and can regulate the physiology of the microvasculature so as to avoid overlarge hydrostatic pressure fluctuation of capillaries and play an important role in determining the peripheral resistance of the organism. When abnormal changes such as slow blood flow, capillary malformation and lumen stenosis occur in microcirculation, microcirculation disturbance of different degrees can occur. Generally, microcirculation disturbance is divided into two categories, namely structural disturbance and functional disturbance, the microcirculation disturbance is closely related to the occurrence and development of cardiovascular and cerebrovascular diseases on the basis of microangiopathy, and the microcirculation disturbance is an important basis and aggravation factor of target organ damage such as heart, brain, kidney and the like, and under the pathological condition, the material exchange of body tissues and the normal function of organs are seriously influenced.
Oyster (Ostrea), also known as fresh oyster, has high nutritive value, high protein content, rich and uniform amino acid content, and is called as "sea milk". Oyster peptide refers to polypeptide obtained by hydrolyzing or degrading protein contained in oysters by chemical or biological methods, and oyster peptide has small relative molecular weight and is easier to absorb and utilize than protein.
The preparation method of the oyster peptide usually adopts an enzymolysis method as a main method, and comprises a single enzymolysis method, a compound enzymolysis method, a multi-enzyme distribution enzymolysis method, an enzymolysis-hydrolysis combined method and the like. The oyster peptides prepared by different methods have different molecular weight distributions and different efficacies. The biological activities of oyster peptide reported at present comprise immunoregulation, antibiosis, antivirus, blood sugar reduction, blood fat reduction, blood pressure reduction, antioxidation, anti-aging and the like, and the multiple effects and the improvement of microcirculation disturbance have a certain relation up to now.
Chinese patent application CN111334549A discloses oyster peptide and an oyster peptide extraction method, wherein fresh shell-free oysters are used as raw materials, the oyster peptide is extracted by enzymolysis of protease special for fresh oyster meat, oyster biological enzyme is hydrolyzed to be completely water-soluble active ingredients, the small peptide content of the product is high, and the taste of the oyster peptide is improved by deodorizing with active carbon, so that the oyster peptide is more beneficial to absorption and utilization of the oyster active ingredients. The oyster peptide and the oyster peptide extraction method have the advantages of simple operation, good protein degradation effect, high oyster peptide yield and higher sensory acceptability, are easy for industrial scale production, have uniform molecular weight distribution, change the oyster into all water-soluble active ingredients after biological enzyme hydrolysis by a biological enzymolysis technology, are completely and quickly absorbed, have high small peptide content in the product, and are more beneficial to the absorption and utilization of the oyster active ingredients; the oyster peptide is more acceptable in taste after being deodorized by active carbon.
Chinese patent application CN109825545A discloses a preparation method of oyster oligopeptide freeze-dried powder, which comprises the following steps: after pretreatment, carrying out enzymolysis on the oyster slurry by adopting alkaline protease, and then carrying out enzymolysis by adopting neutral protease FoodProPNL and flavourzyme LAP; centrifuging the enzymolysis solution, performing ultrafiltration on the obtained supernatant, and then concentrating and performing chromatography on the filtrate; freeze-drying and pulverizing the chromatographed material by vacuum freeze-drying technology. The invention adopts a sustainable step-by-step enzymolysis method, so that the enzymolysis of the oysters is more thorough, the utilization rate of raw materials is improved, and the taste of the product is improved; the invention combines vacuum freeze drying and superfine grinding technologies for preparing oyster oligopeptide, can preserve active ingredients of the oyster oligopeptide and can improve the bioavailability of the oyster oligopeptide.
Chinese patent application CN109504732A discloses a preparation method of oyster active peptide, comprising the following steps: 1) pretreating oyster meat; 2) extracting protein; 3) adding the extracted protein into distilled water according to a feed-liquid ratio of 1: 8-15 g/mL, wherein the distilled water contains protease with the mass concentration of 0.5-0.7%, carrying out enzymolysis for 3-5 hours in a constant-temperature water bath kettle at 45-55 ℃, keeping shaking in the enzymolysis process, heating in a water bath to 90 ℃ for enzyme deactivation, centrifuging, and taking the supernatant to obtain an enzymolysis liquid; 4) precipitating the enzymolysis solution with 70% ethanol solution, and freeze drying to obtain Concha Ostreae active peptide. The oyster active peptide prepared by the method has high purity and strong biological activity, the ratio of small molecular polypeptide with the molecular weight of below 5000Da is up to more than 56.51%, and the oyster active peptide is more beneficial to human body absorption. The oyster active peptide can effectively inhibit alpha-amylase and superoxide anion and effectively remove DPPH, and has good hypoglycemic activity and antioxidant activity.
Disclosure of Invention
The invention aims to provide a preparation method of oyster powder with the effect of improving microcirculation.
The purpose of the invention is realized by the following technical scheme.
A preparation method of oyster powder with microcirculation improving effect comprises the following steps:
s1: adding water into cleaned Carnis Ostreae, and ultrasonic pulverizing to obtain slurry A.
S2: adding n-hexane into the slurry A for extraction and degreasing, and discarding the n-hexane phase to obtain slurry B;
s3: adding arabinose and dry yeast into the slurry B for deodorization to obtain slurry C;
s4: adjusting the pH value of the slurry C to 6-8, adding neutral protease for enzymolysis, inactivating enzyme, and centrifuging to obtain an enzymolysis solution;
s5: concentrating and drying the enzymolysis liquid to obtain polypeptide powder A, and adding an ethanol solution of sulfuric acid for alcoholysis to obtain an alcoholysis liquid;
s6: concentrating the alcoholysis solution, ultrafiltering, drying the filter residue, and grinding to obtain the Concha Ostreae powder.
Further, in S1, the water adding amount is 10-15 times of the weight of the oyster meat, and the ultrasonic frequency is 20-25 Hz.
Further, in S2, n-hexane with the weight 10-15 times that of the oyster meat is added for extraction for 2-3 times.
Further: in S3, arabinose is added in an amount of 1-2% of the oyster meat, and dry yeast is added in an amount of 0.3-0.5% of the oyster meat.
Further, in S4, the amount of neutral protease added to the reaction solution was 1000-1500U/g based on the amount of oyster meat.
Further, in S4, the enzymolysis temperature is 30-40 ℃.
Further, in S4, the enzymolysis time is 5-8 h.
Further, in S4, the end point of the enzymolysis is that the average molecular weight of the polypeptide in the enzymolysis liquid is 3k-5 kDa.
Further, in S5, the mass concentration of sulfuric acid in the ethanol solution of sulfuric acid is 2 to 4%.
Further, in S5, the mass of the ethanol solution of sulfuric acid is 20-30 times of that of the polypeptide powder A.
Further, in S5, the reaction temperature for alcoholysis is 70-90 ℃.
Further, in S5, the reaction time of alcoholysis is 3-5 h.
Further, in S5, the end point of the alcoholysis reaction is that the average molecular weight of the small-molecule peptide in the alcoholysis solution is 0.5k-1 kDa.
Further, in S6, the ultrafiltration uses an ultrafiltration membrane having a molecular weight cut-off of 200-.
Further, in S6, the filter residue was washed with ethanol at-10 to 0 ℃ during ultrafiltration until the filtrate pH was 6 to 7.
The invention has the advantages that:
1. the oyster powder prepared by the preparation method can effectively improve microcirculation disturbance, and has good application prospect in hypertension, diabetes mellitus and microcirculation disturbance caused by occupational environments such as high temperature, noise and the like.
2. The oyster powder prepared by the preparation method of the invention mainly contains micromolecular oyster peptide, is sealed by ethanol, delays the degradation rate after entering the body through oral administration, prolongs the metabolism time in the body, and can play a role for a long time after being taken once.
3. The micromolecule oyster peptide prepared by the preparation method is capped by ethanol, so that the water absorption rate of the product prepared into micromolecule peptide powder can be reduced, and the storage time can be prolonged.
4. The invention adopts two steps of enzymolysis and alcoholysis to compositely treat the oysters, detects the reaction end point by detecting the molecular weight of the enzymolysis reaction and the alcoholysis reaction, and obtains the mixture of a plurality of compounds with small molecular weight, and the compounds are blocked by ethanol, thereby having better effect in resisting microcirculation disturbance.
Detailed Description
The oyster used in the invention is derived from the Crassostrea lactiflora pall of Shandong.
The average molecular weight in the present invention is analyzed and calculated by Gel Permeation Chromatography (GPC).
Example 1
A preparation method of oyster powder with microcirculation improving effect comprises the following steps:
s1: adding 15 times of water into cleaned Carnis Ostreae, and crushing for 2 hr under 20kHz ultrasonic wave to obtain slurry A.
S2: adding n-hexane 15 times the weight of the oyster meat into the slurry A, extracting for 2 times to degrease, and discarding the n-hexane phase to obtain slurry B;
s3: adding 1 wt% of arabinose and 0.3 wt% of dry yeast into the slurry B for deodorization to obtain slurry C;
s4: adjusting the pH value of the slurry C to 6, adding 1000U/g of neutral protease based on the weight of the oyster meat, performing enzymolysis at 35 deg.C for 5h, inactivating enzyme, and centrifuging to obtain an enzymolysis solution; at the moment, the average molecular weight of the polypeptide in the detected enzymolysis liquid is about 4.8 KDa;
s5: concentrating and drying the enzymolysis liquid to obtain polypeptide powder A, adding a 3% sulfuric acid ethanol solution with the mass 20 times that of the polypeptide powder, heating to 80 ℃, reacting for 3 hours to obtain an alcoholysis liquid, and detecting to obtain the average molecular weight of the small molecular peptide of 0.78 kDa;
s6: concentrating the alcoholysis solution, performing ultrafiltration by using an ultrafiltration membrane with the molecular weight cutoff of 300Da, washing filter residues by using ethanol with the temperature of-10 ℃ until the pH of the filtrate is 7, and drying the filter residues to obtain the oyster powder.
Example 2
A preparation method of oyster powder with microcirculation improving effect comprises the following steps:
s1: adding water 10 times the weight of the cleaned Carnis Ostreae into the cleaned Carnis Ostreae, and crushing for 4 hr under 25kHz ultrasonic wave to obtain slurry A.
S2: adding 10 times weight of normal hexane into the slurry A, extracting for 3 times to defat, and discarding normal hexane phase to obtain slurry B;
s3: adding 1 wt% of arabinose and 0.5 wt% of dry yeast into the slurry B for deodorization to obtain slurry C;
s4: adjusting the pH value of the slurry C to 6, adding 1300U/g of neutral protease based on the weight of the oyster meat, performing enzymolysis at 35 deg.C for 8h, inactivating enzyme, and centrifuging to obtain an enzymolysis solution; at the moment, the average molecular weight of the polypeptide in the detected enzymolysis liquid is about 3.3 KDa;
s5: concentrating and drying the enzymolysis liquid to obtain polypeptide powder A, adding a 4% sulfuric acid ethanol solution with the mass 30 times that of the polypeptide powder, heating to 80 ℃, reacting for 5 hours to obtain an alcoholysis liquid, and detecting to obtain the average molecular weight of the small molecular peptide of 0.53 kDa;
s6: concentrating the alcoholysis solution, performing ultrafiltration by using an ultrafiltration membrane with the molecular weight cutoff of 200Da, washing filter residues by using ethanol at 0 ℃ until the pH value of the filtrate is 6, and drying the filter residues to obtain the oyster powder.
Example 3
A preparation method of oyster powder with microcirculation improving effect comprises the following steps:
s1: adding 12 times of water into cleaned Carnis Ostreae, and pulverizing for 3 hr under 22kHz ultrasonic wave to obtain slurry A.
S2: adding n-hexane 15 times the weight of the oyster meat into the slurry A, extracting for 2 times to degrease, and discarding the n-hexane phase to obtain slurry B;
s3: adding 1 wt% of arabinose and 0.3 wt% of dry yeast into the slurry B for deodorization to obtain slurry C;
s4: adjusting the pH value of the slurry C to 8, adding 1500U/g neutral protease based on the weight of the oyster meat, performing enzymolysis at 35 deg.C for 5h, inactivating enzyme, and centrifuging to obtain enzymolysis solution; at the moment, the average molecular weight of the polypeptide in the detected enzymolysis liquid is about 4.2 KDa;
s5: concentrating and drying the enzymolysis liquid to obtain polypeptide powder A, adding 2% sulfuric acid ethanol solution with the mass 25 times that of the polypeptide powder, heating to 70 ℃, reacting for 3 hours to obtain an alcoholysis liquid, and detecting to obtain the average molecular weight of the small molecular peptide of 0.96 kDa;
s6: concentrating the alcoholysis solution, performing ultrafiltration by using an ultrafiltration membrane with the molecular weight cutoff of 200Da, washing filter residues by using ethanol at 0 ℃ until the pH value of the filtrate is 6, and drying the filter residues to obtain the oyster powder.
Comparative example 1
The difference from example 1 is that in S4, the amount and duration of action of protease are increased, the average molecular weight of peptide in the enzymatic hydrolysate is decreased, and there is no alcoholysis step.
A preparation method of oyster powder comprises the following steps:
s1: adding 15 times of water into cleaned Carnis Ostreae, and crushing for 2 hr under 20kHz ultrasonic wave to obtain slurry A.
S2: adding n-hexane 15 times the weight of the oyster meat into the slurry A, extracting for 2 times to degrease, and discarding the n-hexane phase to obtain slurry B;
s3: adding 1 wt% of arabinose and 0.3 wt% of dry yeast into the slurry B for deodorization to obtain slurry C;
s4: adjusting the pH value of the slurry C to 6, adding 5000U/g of neutral protease based on the weight of the oyster meat, performing enzymolysis at 35 deg.C for 5h, inactivating enzyme, and centrifuging to obtain an enzymolysis solution; the average molecular weight of the polypeptide in the detected enzymolysis liquid is about 0.73 kDa;
s5: concentrating the enzymolysis liquid, performing ultrafiltration by using an ultrafiltration membrane with the molecular weight cutoff of 300Da, washing filter residues by using ethanol with the temperature of-10 ℃ until the pH value of the filtrate is 7, and drying the filter residues to obtain the oyster powder.
Comparative example 2
The difference from example 1 is that no enzymatic hydrolysis is used, only an alcoholysis step.
A preparation method of oyster powder comprises the following steps:
s1: adding 15 times of water into cleaned Carnis Ostreae, and crushing for 2 hr under 20kHz ultrasonic wave to obtain slurry A.
S2: adding n-hexane 15 times the weight of the oyster meat into the slurry A, extracting for 2 times to degrease, and discarding the n-hexane phase to obtain slurry B;
s3: adding 1 wt% of arabinose and 0.3 wt% of dry yeast into the slurry B for deodorization to obtain slurry C;
s4: concentrating and drying the slurry C to obtain polypeptide powder A, adding a 3% sulfuric acid ethanol solution with the mass 20 times that of the polypeptide powder, heating to 80 ℃, reacting for 3 hours to obtain an alcoholic solution, and detecting to obtain the average molecular weight of the small molecular peptide more than 8 kDa;
s5: concentrating the alcoholysis solution, performing ultrafiltration by using an ultrafiltration membrane with the molecular weight cutoff of 300Da, washing filter residues by using ethanol with the temperature of-10 ℃ until the pH of the filtrate is 7, and drying the filter residues to obtain the oyster powder.
Comparative example 3
The difference from example 1 is that the parameters used in S4 and S5 are different.
A preparation method of oyster powder comprises the following steps:
s1: adding 15 times of water into cleaned Carnis Ostreae, and crushing for 2 hr under 20kHz ultrasonic wave to obtain slurry A.
S2: adding n-hexane 15 times the weight of the oyster meat into the slurry A, extracting for 2 times to degrease, and discarding the n-hexane phase to obtain slurry B;
s3: adding 1 wt% of arabinose and 0.3 wt% of dry yeast into the slurry B for deodorization to obtain slurry C;
s4: adjusting the pH value of the slurry C to 5, adding 1000U/g of neutral protease based on the weight of the oyster meat, performing enzymolysis at 35 deg.C for 3h, inactivating enzyme, and centrifuging to obtain enzymolysis solution; at the moment, the average molecular weight of the polypeptide in the detected enzymolysis liquid is about 7.3 KDa;
s5: concentrating and drying the enzymolysis liquid to obtain polypeptide powder A, adding a 2% sulfuric acid ethanol solution with the mass 20 times that of the polypeptide powder, heating to 80 ℃, reacting for 3 hours to obtain an alcoholysis liquid, and detecting to obtain the average molecular weight of the small molecular peptide of 2.6 kDa;
s6: concentrating the alcoholysis solution, performing ultrafiltration by using an ultrafiltration membrane with the molecular weight cutoff of 300Da, washing filter residues by using ethanol with the temperature of-10 ℃ until the pH of the filtrate is 7, and drying the filter residues to obtain the oyster powder.
The efficacy of improving microcirculation is verified by an adrenaline hydrochloride-induced microcirculation disturbance animal model.
Experimental animals: the SPF-level Kunming-breed mice are half female and half male, and have the body mass of 18-22 g.
The experimental method comprises the following steps: the group was divided into a blank control group, a model group, example 1, comparative example 2 and comparative example 3, wherein the blank control group and the model group were fed with normal feed while 1mL of distilled water was gavaged at the same time every day. Example 1, comparative example 2 and comparative example 3 groups of mice were fed normal diet, while the product of the respective examples was gavaged daily at the same time at a dose of 1g/kg BW for 15 days. Mice were injected intraperitoneally with 5% chloral hydrate (3 ml. kg) 1h after gavage on day 15-1) Anaesthetizing, fixing anesthetized mouse on mouse observation table with abdomen facing downwards, adjusting ear support height to make auricle flat on ear support, dripping small amount of cedar oil on auricle surface, placing observation table on microscope stage, observing auricle arteriole, venule diameter and capillary vessel opening number of each mouse with medical image analysis system (BI-2000 medical image analysis system), and immediately injecting AD (adrenaline hydrochloride) into tail vein of other groups except blank control group) Injection (10mg kg)-1) After 5min, the mice were observed for the number of arterioles, venules and capillaries opened. The effect of each group of products on resisting microcirculation disturbance is judged according to the change of the vascular state before and after AD injection. The smaller the change of the vascular state before and after AD injection, the better the effect. The results are shown in Table 1.
TABLE 1 vascular changes in mice of the experimental groups: (
n＝10)
P < 0.05, P < 0.01, compared to model control.
It can be seen that, in the case that the mouse continuously intakes the product, in the microcirculation disturbance modeling experiment, the experimental group which intakes the oyster powder of the application has small change of the vascular state after modeling, the vascular state after modeling is closer to the normal group, and has very significant difference in every aspect with the model control group. After the model is made, the vascular state of each group of the comparative example is greatly different from that of the normal group, and compared with the model group, the vascular state cannot achieve the effect of extremely significant difference in all aspects, so that the comparative example has poor resistance to microcirculation disturbance.
Finally, it should be noted that the above-mentioned contents are only used for illustrating the technical solutions of the present invention, and not for limiting the protection scope of the present invention, and that the simple modifications or equivalent substitutions of the technical solutions of the present invention by those of ordinary skill in the art can be made without departing from the spirit and scope of the technical solutions of the present invention.
Claims (10)
1. A preparation method of oyster powder with microcirculation improving effect comprises the following steps:
s1: adding water into the cleaned oyster meat, and carrying out ultrasonic crushing to obtain slurry A;
s2: adding n-hexane into the slurry A for extraction and degreasing, and discarding the n-hexane phase to obtain slurry B;
s3: adding arabinose and dry yeast into the slurry B for deodorization to obtain slurry C;
s4: adjusting the pH value of the slurry C to 6-8, adding neutral protease for enzymolysis, inactivating enzyme, and centrifuging to obtain an enzymolysis solution;
s5: concentrating and drying the enzymolysis liquid to obtain polypeptide powder A, and adding an ethanol solution of sulfuric acid for alcoholysis to obtain an alcoholysis liquid;
s6: concentrating the alcoholysis solution, ultrafiltering, drying the filter residue, and grinding to obtain the Concha Ostreae powder.
2. The preparation method according to claim 1, wherein in S1, the amount of water added is 10-15 times of the weight of the oyster meat, and the ultrasonic frequency is 20-25 Hz; in S2, n-hexane which is 10-15 times of the weight of the oyster meat is added for extraction for 2-3 times.
3. The method according to claim 1, wherein in S3, arabinose is added in an amount of 1-2% and dry yeast is added in an amount of 0.3-0.5% based on the weight of the oyster meat.
4. The method according to claim 1, wherein the amount of neutral protease added in the reaction solution of S4 is 1000-1500U/g based on oyster meat.
5. The preparation method according to claim 1, wherein in S4, the enzymolysis temperature is 30-40 ℃, and the enzymolysis time is 5-8 h.
6. The method according to claim 1, wherein the end point of the enzymatic hydrolysis in S4 is that the average molecular weight of the polypeptide in the enzymatic hydrolysate is 3k-5 kDa.
7. The preparation method according to claim 1, wherein in S5, the mass concentration of sulfuric acid in the ethanol solution of sulfuric acid is 2-4%; the mass of the ethanol solution of the sulfuric acid is 20-30 times of that of the polypeptide powder A.
8. The method according to claim 1, wherein in S5, the reaction temperature for alcoholysis is 70-90 ℃; the reaction time of alcoholysis is 3-5 h.
9. The method according to claim 1, wherein in S5, the alcoholysis reaction is terminated at a point where the average molecular weight of the small-molecule peptide in the alcoholysis solution is 0.5k-1 kDa.
10. The method according to claim 1, wherein in S6, the ultrafiltration uses an ultrafiltration membrane having a molecular weight cut-off of 200-300; during the ultrafiltration, the filter residue is washed by ethanol at the temperature of-10-0 ℃ until the pH value of the filtrate is 6-7.
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