CN111802653A - Tuna oligopeptide powder composition and preparation method and application thereof - Google Patents
Tuna oligopeptide powder composition and preparation method and application thereof Download PDFInfo
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- CN111802653A CN111802653A CN202010957587.0A CN202010957587A CN111802653A CN 111802653 A CN111802653 A CN 111802653A CN 202010957587 A CN202010957587 A CN 202010957587A CN 111802653 A CN111802653 A CN 111802653A
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- enzymolysis
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- oligopeptide powder
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Abstract
The invention relates to a tuna oligopeptide powder composition, which comprises the following components in parts by weight: 25-30 parts of tuna oligopeptide powder, 10-15 parts of fructo-oligosaccharide, 15-20 parts of fish collagen peptide powder, 15-20 parts of acerola juice powder, 15-20 parts of juicy peach juice powder, 5-10 parts of erythritol, 1-2 parts of compound sour sweetener and 3-4 parts of water-soluble sugar; the water-soluble sugar is fucoidin or chitosan oligosaccharide. The tuna oligopeptide powder composition is used for reducing the uric acid level in serum. The tuna oligopeptide powder composition can effectively reduce the serum uric acid level, the main component of the tuna oligopeptide powder composition is easy to absorb by a human body, various raw materials and auxiliary materials are matched to help to maintain the health level, and the tuna oligopeptide powder composition has no toxic or side effect after being eaten for a long time, can effectively overcome the defect of side effect of medicines, and can maintain the health condition of the human body.
Description
Technical Field
The invention relates to a tuna oligopeptide powder composition, a preparation method and application thereof, and belongs to the technical field of marine fish food processing.
Background
In recent years, the incidence of gout has been on the rise year by year with changes in factors such as lifestyle, eating habits, and the surrounding environment. Gout is a metabolic disease caused by purine metabolic disorder of human body, and hyperuricemia is the biochemical basis of gout. Uric acid is a final product of purine substance metabolism of a human body, is slightly soluble in the blood environment of the body, and when the production amount of uric acid is far higher than the excretion amount, the blood uric acid level in the body is continuously increased and is higher than the solubility to form sodium urate crystals, the sodium urate crystals are deposited on tissues such as joints, kidneys and the like to cause inflammation, and severe cases form tophus, and finally gout occurs.
The control of uric acid level is the key to gout treatment, and has important significance for preventing gout, repeated gout attack and disease deterioration. Western medicines such as colchicine, non-steroidal anti-inflammatory drugs, prednisolone, allopurinol and the like are commonly used for treating hyperuricemia or gout at present. Although the western medicine has quick response, certain side effects exist, such as kidney and liver dysfunction, abnormal carbohydrate metabolism, central nervous system damage, muscular atrophy and the like.
Taking allopurinol which is specially used for resisting the generation of uric acid as an example, the allopurinol is clinically and specially used for diseases such as primary and secondary hyperuricemia, recurrent gout, tophus and the like. However, allopurinol can cause various adverse reactions, such as gastrointestinal reactions such as pruritic papules or urticaria, diarrhea and vomiting and the like, leukopenia, alopecia, hypodynamia, numbness of hands and feet, lymphadenectasis and the like, and even unknown death cases appear in foreign countries and even during the medicine taking period. In view of the toxic and side effects of allopurinol, the dosage needs to be strictly controlled in the process of taking, and the medicine is taken according to the medical advice strictly aiming at specific conditions.
Aiming at the current situation that western medicines for treating hyperuricemia generally have obvious adverse reactions, the development of the high-efficiency low-toxicity uric acid reducing active substance has important significance.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides the tuna oligopeptide powder composition and the preparation method and the application thereof, the tuna oligopeptide powder composition can effectively reduce the serum uric acid level, the main component of the tuna oligopeptide powder composition is easy to be absorbed by human bodies, various raw and auxiliary materials are matched in a compatible manner, the health level is maintained, no toxic or side effect is caused after long-term eating, and the defect of side effect of medicines can be effectively overcome.
The technical scheme for solving the technical problems is as follows: the tuna oligopeptide powder composition comprises the following components in parts by weight: 25-30 parts of tuna oligopeptide powder, 10-15 parts of fructo-oligosaccharide, 15-20 parts of fish collagen peptide powder, 15-20 parts of acerola juice powder, 15-20 parts of juicy peach juice powder, 5-10 parts of erythritol, 1-2 parts of compound sour sweetener and 3-4 parts of water-soluble sugar; the water-soluble sugar is fucoidin or chitosan oligosaccharide.
Preferably, the content of the tuna oligopeptide functional dipeptide in the tuna oligopeptide powder composition is 20-60 mg.
Preferably, the compound sour sweetener comprises DL-malic acid, citric acid, sodium citrate and a sweetener, the weight ratio of the DL-malic acid to the citric acid to the sodium citrate to the sweetener is 3:2:1:1, and the sweetener is sucralose, mogroside or stevioside.
Preferably, the powder size of the tuna oligopeptide powder composition is 60-100 meshes.
Tuna oligopeptide powder: the tuna meat is prepared by a biological enzymolysis process by taking tuna meat as a raw material, has high nutritional value, contains more than 85% of protein, contains 20 amino acids required by a human body, 12.1% of branched chain amino acid and 1.3% of taurine, and is rich in functional components such as carnosine and anserine and mineral elements such as calcium, zinc and selenium; the molecular weight is moderate, the average relative molecular mass is 607Da, the contents of dipeptide and tripeptide are 45.77%, and the content of polypeptide with the relative molecular mass less than 1000Da is 89.83%; the absorption and utilization rate is high, the amino acid proportioning mode is close to that of a human body, the relative molecular mass is small, digestion is not needed, and the amino acid is directly absorbed in small intestines in an integral form, participates in metabolism of the human body and exerts the biological activity function of the amino acid. Besides the function of reducing uric acid level, the tuna oligopeptide also has the functions of immunoregulation, fatigue resistance, oxidation resistance, blood pressure reduction and the like. The main functions of the tuna oligopeptide powder in the formula are to reduce the uric acid level of a patient with high uric acid and the high urea and creatinine levels caused by high uric acid, and no related report that the tuna oligopeptide powder has the function of reducing uric acid exists at present.
Fish collagen peptide powder: the protein-rich haydite is prepared by taking alas deepened sea cod skin as a raw material through a biological enzymolysis process, the protein content is up to 95 percent, and the average relative molecular mass is 705 Da; is rich in small molecule active peptide, amino acid and other components, can be directly absorbed and utilized without digestion, and has the absorption utilization rate as high as 90 percent. The fish collagen peptide has the functions of immunoregulation, antioxidation, bone joint health maintenance and the like.
Fucoidin: fucoidin is a unique water-soluble polysaccharide combined with sulfuric acid groups, and has the physiological functions of regulating immunity, eliminating free radicals, resisting aging, reducing blood fat, reducing blood pressure and the like. A large number of clinical studies prove that hyperuricemia is easy to induce gout, most hyperuricemia patients are accompanied by hypertension, renal function injury and the like with different degrees, and fucoidan has the function of relieving chronic renal failure, so that renal function is effectively protected. The fucoidin is added in the formula, so that adverse reactions such as renal function injury and the like of hyperuricemia patients caused by metabolic disorder are relieved, and the fucoidin is coordinated with tuna oligopeptide to jointly improve the metabolic condition of the patients and maintain the health level.
Chitosan oligosaccharide: the chitosan oligosaccharide is a micromolecular oligosaccharide with amino groups degraded by chitosan derived from shrimp and crab shells, is the only alkaline amino oligosaccharide with positive charges in the nature, and is animal dietary fiber and prebiotics. The polymerization degree of the chitosan oligosaccharide is between 2 and 20, the water solubility is good, the biological activity is high, and the chitosan oligosaccharide has the functions of assisting in reducing the uric acid level, regulating the immunity, improving the intestinal micro-ecology and the like. The compatibility of the tuna oligopeptide powder in the formula is beneficial to reducing the serum uric acid level; the fructo-oligosaccharide serving as animal prebiotics is compatible with plant prebiotics and is beneficial to absorption and utilization; and the chitosan oligosaccharide is beneficial to improving the taste of a formula sample and is easy to accept.
The acerola juice powder is rich in anthocyanin, melatonin, quercetin and other components, and has the functions of resisting inflammation, easing pain, resisting tumor, resisting oxidation, reducing blood sugar, protecting nerve cells and the like. The tuna oligopeptide and the fish collagen peptide are mutually compatible in the formula, so that the absorption and utilization are facilitated, and the harm of hyperuricemia to human bodies is resisted together.
The honey peach juice powder is rich in vitamin C, is sour, sweet, tasty, fresh and fine, can effectively cover the fishy smell of the tuna oligopeptide, brings the taste of the fruit drink, is easy to accept by consumers, and can be eaten for a long time.
The fructo-oligosaccharide is a natural active substance, maintains the pure sweet taste of sucrose, has refreshing sweet taste compared with sucrose, has the functions of regulating intestinal flora, proliferating bifidobacteria, promoting calcium absorption, regulating blood lipid, regulating immunity and the like, does not participate in sugar metabolism, and can be eaten by diabetic patients.
Erythritol is a zero-calorie sweetener, does not participate in sugar metabolism and blood sugar change, has a cooling feeling, can cover up the bad taste of the high-power sweetener, and is beneficial to improving the overall taste of a formula product.
The ingredients of the compound sour sweetener are blended according to a certain proportion and then added into the product, which has great influence on the flavor and taste of the product and plays a role in adjusting the taste.
Besides the main functional components such as tuna oligopeptide powder, fucoidin, chitosan oligosaccharide, acerola cherry juice powder and the like, the types and the amounts of the raw and auxiliary materials can be adjusted according to the specific formula characteristics or requirements, so that the actual effect of reducing the uric acid level of the formula composition product is not influenced.
The invention also discloses a preparation method of the tuna oligopeptide powder composition, which comprises the following steps:
1) preparation of tuna oligopeptide powder:
(a) pretreatment of raw materials: thawing raw water of tuna, taking tuna meat, washing twice with tap water, and washing once with purified water; (b) crushing: pulverizing tuna meat for 30min with an automatic chopper mixer to obtain homogenate; (c) pressure heat treatment: pouring the homogenate into a pressure stirring pot, adding purified water with the mass 2 times that of the fish meat, heating to 120 ℃, preserving heat for 2 hours, closing the pressure stirring pot, and naturally cooling to normal temperature and normal pressure; (d) enzymolysis: transferring the homogenate after the heat treatment into an enzymolysis tank for enzymolysis to obtain an enzymolysis liquid; (c) centrifuging: heating the enzymolysis solution to 60 ℃, filtering the enzymolysis solution by a 100-mesh filter screen, standing for 2 hours, then feeding the enzymolysis solution into a centrifugal machine, centrifuging at 6900r/min, feeding at a speed of 0.5t/h, discharging slag for 10 min/time, and feeding the centrifuged feed liquid into a liquid storage tank; (d) and (3) decoloring: heating the feed liquid to 50 ℃, adding active carbon accounting for 5% of the fish meat mass, and stirring for 30 min; (e) and (3) filtering: filter pressing the feed liquid for 2 times by using diatomite as a filter cake; (f) concentration and purification: pumping the filtrate into a liquid storage tank, and circulating through an ultrafiltration device, wherein the cut-off molecular weight of the ultrafiltration membrane is 10000Da, and the pressure is lower than 0.2 MPa; nano-filtering, concentrating and desalting to remove 80% of water; transferring the feed liquid into a single-effect evaporator, concentrating for the second time until the concentration of the feed liquid is 12-16%, controlling the pressure to be 0.08MPa, discharging evaporation condensate water every half hour, and heating to 80 ℃; (g) spray drying: preheating spray drying equipment, heating the feed liquid to 90 ℃, pumping the feed liquid into a spray drying tower, controlling the temperature at 150 ℃, the feeding amount at 50kg/h and the powder collecting frequency at 1 h/time; (h) and (3) packaging and storing: putting the dried powder into a food-grade plastic bag, fastening the opening of the bag, labeling, and storing in a warehouse;
the preparation method of the tuna oligopeptide powder comprises the steps of firstly screening various parts of tuna raw materials, respectively carrying out biological enzymolysis on tuna skin, tuna head, tuna bone and tuna meat to prepare the oligopeptide powder, measuring the content of functional dipeptide by using a high performance liquid chromatography, and selecting the tuna meat as the raw material by combining the measurement result and the actual production process condition.
2) Preparing fish collagen peptide powder:
(a) pretreatment of raw materials: cleaning Alaska deepened sea cod skin twice with tap water, and cleaning with purified water once; (b) crushing: crushing the cod skin for 10min by using an automatic cutting and mixing machine to obtain homogenate; (c) pressure heat treatment: pouring the homogenate into a pressure stirring pot, adding purified water 2 times the weight of the fish skin, heating to 120 ℃, keeping the temperature for 30min, closing the pressure stirring pot, and naturally cooling to normal temperature and normal pressure; (d) enzymolysis: transferring the homogenate after the heat treatment into an enzymolysis tank for enzymolysis to obtain an enzymolysis liquid; (e) centrifuging: heating the enzymolysis solution to 60 ℃, filtering the enzymolysis solution by a 100-mesh filter screen, standing for 2 hours, then feeding the enzymolysis solution into a centrifugal machine, centrifuging at 6900r/min, feeding at a speed of 0.5t/h, discharging slag for 10 min/time, and feeding the centrifuged feed liquid into a liquid storage tank; (f) and (3) decoloring: heating the feed liquid to 50 ℃, adding active carbon accounting for 5% of the weight of the fish skin, and stirring for 30 min; (g) and (3) filtering: filter pressing the feed liquid for 2 times by using diatomite as a filter cake; (h) concentration and purification: pumping the filtrate into a liquid storage tank, circulating through an ultrafiltration device, intercepting the molecular weight of the ultrafiltration membrane to 10000Da and concentrating to remove 80% of water, wherein the pressure is lower than 0.2 MPa; transferring the feed liquid into a single-effect evaporator, concentrating for the second time until the concentration of the feed liquid is 12-16%, controlling the pressure to be 0.08MPa, discharging evaporation condensate water every half hour, and heating to 80 ℃; (i) spray drying: preheating spray drying equipment, heating the feed liquid to 90 ℃, pumping the feed liquid into a spray drying tower, controlling the temperature at 150 ℃, the feeding amount at 50kg/h and the powder collecting frequency at 1 h/time; (j) and (3) packaging and storing: putting the dried powder into a food-grade plastic bag, fastening the opening of the bag, labeling, and storing in a warehouse;
other raw and auxiliary materials are powder products purchased from the market, and all the raw and auxiliary materials are food production permission qualifications.
3) Tuna oligopeptide powder composition:
the tuna oligopeptide powder composition comprises the following components in parts by weight: 25-30 parts of tuna oligopeptide powder, 10-15 parts of fructo-oligosaccharide, 15-20 parts of fish collagen peptide powder, 15-20 parts of acerola juice powder, 15-20 parts of juicy peach juice powder, 5-10 parts of erythritol, 1-2 parts of compound sour sweetener and 3-4 parts of water-soluble sugar; the water-soluble sugar is fucoidin or chitosan oligosaccharide;
weighing the raw materials in proportion, sequentially sieving with 60 mesh sieve, transferring the sieved raw materials into a V-shaped mixer, starting a power supply to automatically mix for 35min, transferring the uniformly mixed powder to a full-automatic quantitative packaging machine, adjusting the metering value to 4 g/bag, and automatically packaging. The automatically packaged products are packed in boxes, the specification is 30 bags/box, one box is used for one period, the serum uric acid level of patients with hyperuricemia can be effectively reduced, and the body feeling effect is good.
Further, in the preparation process of the tuna oligopeptide powder, the enzymolysis process is as follows: transferring the heat-treated homogenate into an enzymolysis tank, adding purified water with the mass 8 times that of fish, stirring uniformly, adjusting the temperature to 50-52 ℃, adjusting the pH to =9, adding alkaline protease with the mass 0.5% of fish for enzymolysis for 2h, adjusting the pH of the homogenate to 7, sequentially adding neutral protease, compound protease and flavor enzyme with the mass 0.5% of fish, performing enzymolysis for 1h, controlling the temperature to 50-52 ℃ in the enzymolysis process, and stirring once every 10min to obtain an enzymolysis solution.
Further, in the preparation process of the fish collagen peptide powder, the enzymolysis process is as follows: transferring the heat-treated homogenate into an enzymolysis tank, adding purified water 6 times the mass of the fish skin, uniformly stirring, adjusting the temperature to 53-55 ℃, adjusting the pH to =9, adding alkaline protease 0.5% of the mass of the fish skin for enzymolysis for 2h, adjusting the pH of the homogenate to 7, sequentially adding neutral protease 0.5% of the mass of the fish skin and flavor enzyme, performing enzymolysis for 1h, controlling the temperature to 53-55 ℃ in the enzymolysis process, and stirring once every 10 min.
The invention also discloses an application of the tuna oligopeptide powder composition, and the tuna oligopeptide powder composition is used for reducing the uric acid level in serum.
The invention has the beneficial effects that: the tuna oligopeptide powder composition can reduce the uric acid level of a patient, effectively relieve hyperuricemia symptoms, can effectively relieve renal function injury caused by uric acid metabolic disorder by adding fucoidin in an auxiliary manner, is also favorable for reducing serum uric acid level by matching chitosan oligosaccharide added in the auxiliary manner with the tuna oligopeptide powder, and is more favorable for absorption and utilization by matching the chitosan oligosaccharide and fructo-oligosaccharide. In addition, the tuna oligopeptide powder and the fish collagen peptide powder are small molecular peptides with biological activity, have the functions of resisting oxidation, regulating immunity and the like, are beneficial to regulating various functions and promote the health of human bodies.
In addition, the juicy peach juice powder, acerola cherry juice powder and various sour sweeteners are added into the formula product to adjust the taste, the special taste of the tuna oligopeptide powder is covered, the whole taste of the formula is fresh and cool, the tuna oligopeptide powder is easy to accept, the health level can be maintained due to the compatibility and matching of various raw and auxiliary materials, no toxic or side effect is caused after long-term eating, the defect of side effect of the medicine can be effectively overcome, and various raw and auxiliary materials are processed and prepared into convenient food through a food processing technology, so that the instant food is easy to store and carry and is convenient for patients to take for a long.
The active peptide product obtained by the biological enzymolysis technology belongs to food and has no toxic or side effect on human bodies after being eaten for a long time. Moreover, the small molecular peptide is easier to absorb, has the functions of reducing blood pressure, cholesterol, thrombus and the like, can effectively improve the utilization value of the marine fish, can expand the source range of active substances for reducing uric acid, and fills up the blank of the current market.
Drawings
FIG. 1 comparison of the body weight changes of the mice in example 3
）；
FIG. 2 comparison of biochemical index changes in blood of mice in each group in example 3: (
）；
(A) Uric Acid (UA), (B) Creatinine (CREA), (C) Urea (Urea), (D) blood Glucose (GLU), (E) Total Cholesterol (TCH), (F) triglyceride (TG-SH), (G) alanine Aminotransferase (ALT), (H) aspartate Aminotransferase (AST), (I) Adenosine Deaminase (ADA); (P < 0.05;. P < 0.01);
FIG. 3 comparison of the content of Xanthine Oxidase (XOD) in the serum of mice of each group in example 3: (
）（*P＜0.05；**P＜0.01）；
FIG. 4 the liver Adenosine Deaminase (ADA) and Xanthine Oxidase (XOD) contents of each group of mice in example 3: (
）（*P＜0.05；**P＜0.01）；
FIG. 5 comparison of liver XOD and ADA m RNA expression in each group of mice in example 3: (
）（*P＜0.05；**P＜0.01）
Detailed Description
The present invention will be described in detail with reference to the following embodiments in order to make the aforementioned objects, features and advantages of the invention more comprehensible. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.
Example 1
Preparation of tuna oligopeptide powder composition:
the tuna oligopeptide powder composition comprises the following components in parts by weight: 26 parts of tuna oligopeptide powder, 16.8 parts of fish collagen peptide powder, 16 parts of acerola juice powder, 15 parts of honey peach juice powder, 10 parts of fructo-oligosaccharide, 10 parts of erythritol, 3 parts of fucoidin and 1.2 parts of compound sour sweetener.
The compound sour sweetener is prepared by blending DL-malic acid, citric acid, sodium citrate and sucralose according to the proportion of 3:2:1: 1.
Weighing the raw and auxiliary materials according to a certain proportion, sequentially sieving the raw and auxiliary materials by a 60-mesh sieve, transferring the sieved raw and auxiliary materials into a V-shaped mixer, starting a power supply to automatically mix for 35min, transferring the uniformly mixed powder to a full-automatic quantitative packaging machine, adjusting the metering value to 4 g/bag, and automatically packaging. The automatically packaged products are packed in boxes, the specification is 30 bags/box, one box is used for one period, the serum uric acid level of patients with hyperuricemia can be effectively reduced, and the body feeling effect is good.
Example 2
Preparation of tuna oligopeptide powder composition:
the tuna oligopeptide powder composition comprises the following components in percentage by weight: 25 parts of tuna oligopeptide powder, 16.5 parts of fish collagen peptide powder, 16.25 parts of acerola juice powder, 15 parts of honey peach juice powder, 12.5 parts of fructo-oligosaccharide, 7.5 parts of erythritol, 3 parts of chitosan oligosaccharide and 1.0 part of compound sour sweetener.
The compound sour sweetener is prepared by blending DL-malic acid, citric acid, sodium citrate and sucralose according to the proportion of 3:2:1: 1.
Weighing the raw and auxiliary materials according to a certain proportion, sequentially sieving the raw and auxiliary materials by a 60-mesh sieve, transferring the sieved raw and auxiliary materials into a V-shaped mixer, starting a power supply to automatically mix for 35min, transferring the uniformly mixed powder to a full-automatic quantitative packaging machine, adjusting the metering value to 4 g/bag, and automatically packaging. The automatically packaged products are packed in boxes, the specification is 30 bags/box, one box is used for one period, the serum uric acid level of patients with hyperuricemia can be effectively reduced, and the body feeling effect is good.
Example 3
The influence of tuna oligopeptide powder on the content of uric acid in the serum of a hyperuricemia mouse is as follows:
experimental animals: kunming mice, SPF grade, 72 males, weight 20-22 g, purchased from Jinan Pengyue laboratory animal Breeding Co., Ltd, production license number: SCXK (lu) 20140007.
Reagent consumables: potassium Oxonate (batch: B22D9D 78194), hypoxanthine (batch: Y15J9C 52922), allopurinol (batch: X27J10Y 91606) were purchased from leaf organisms of origin; sodium carboxymethylcellulose (batch: 20160602) and chloroform (batch: 20180808) were purchased from national pharmaceutical group chemical reagents, Inc.; XOD (batch: 01/2020) ELISA kit, Jianglai organism; RevertAId First Strand cDNA Synthesis Kit (batch: 00775466), U.S.A. ThermoFisher; isopropyl alcohol (batch: F715BA 1004), Shanghai Biotech; TRIzol (batch # BCBX 0488), Sigma in USA; chamq SYBR qPCR Master Mix (batch No. 7E381B 9), Nanjing Novowed Biotech, Inc.; DEPC treated Water (batch: 20191206), Solebao.
An experimental instrument:
autoclave (model: GI54 TW), Youmi (Xiamen) instruments, Inc.;
one-ten-thousandth balance (model: ME 204T), Mettler-Torlo;
electronic balance (model: YP 20K), shanghai zhuojing electronics technologies ltd;
microplate reader (model: Elx 808), Berton instruments Inc., USA;
real-time fluorescent quantitative PCR instrument (model: ABI 7500), applied biosystems of America (ABI); ultramicro spectrophotometer (model: DS-11FX +), Demovix, USA.
The experimental method comprises the following steps: a hyperuricemia mouse Model is constructed by feeding hypoxanthine (300 mg/kg) + oxonate potassium (250 mg/kg), SPF-grade Kunming male mice are randomly divided into a blank control group (Normal), a Model group (Model), an allopurinol group (Positive) (40 mg/kg), a tuna oligopeptide powder Low dose group (Low) (286 mg/kg), a tuna oligopeptide powder medium dose group (Middle) (572 mg/kg) and a tuna oligopeptide powder High dose group (High) (858 mg/kg), the mice are perfused for 30d according to 20ml/kg for 1 time every day during the experiment period, the general state of the mice is observed, the weight of the mice is weighed after administration every week, the mice are euthanized after the experiment is finished, blood is taken and centrifuged to obtain serum, and then Uric Acid (UA), Creatinine (CREA), Urea (Urea), blood sugar (GLU), Total Cholesterol (TCH) and Total Cholesterol (TCH) are detected, The content of triglyceride (TG-SH), alanine Aminotransferase (ALT), aspartate Aminotransferase (AST), Adenosine Deaminase (ADA), and Xanthine Oxidase (XOD); collecting liver at low temperature, and detecting the content of Adenosine Deaminase (ADA) and Xanthine Oxidase (XOD) and the expression of liver XOD and ADA mRNA in liver tissue.
The experimental results are as follows:
(i) observation of general conditions in mice
In the whole experiment process, the mice in the blank group had normal drinking water, bright hair, normal stool and good mental status. In the model group and the positive group, some mice have dry fur, no luster, poor mental status and the like at the later stage of the experiment, particularly in the positive control group, the body gradually becomes thin along with the prolonging of the administration time, and 2 mice die at the 28 th day and the 29 th day of the administration. Compared with the model group, the mice in the administration group had regular hair and good mental status.
In the whole experiment process, except for the positive control group, the weight average of the mouse bodies of other groups is in a gradually rising trend, and no significant difference exists; the positive control group gradually decreased in body weight and decreased significantly in the first week and recovered in the second week, but overall decreased in body weight, possibly with damage to mice due to allopurinol side effects (P < 0.01). The specific contents are as follows in table 1:
TABLE 1 comparison of the body weight changes of the mice in the groups: (
）
（**P＜0.01）
(ii) Biochemical index change of blood of mouse
Compared with a blank control group, the serum of the model group mice is obviously increased by uric acid, creatinine and adenosine deaminase (P is less than 0.01); blood sugar and alanine aminotransferase are obviously reduced; urea, total cholesterol, triglycerides, aspartate aminotransferase were not significantly changed. The serum contents of uric acid (P < 0.01) and adenosine deaminase (P < 0.05) in the positive control group are obviously reduced compared with the serum contents in the model control group; serum creatinine and urea were significantly elevated (× P < 0.01), suggesting that renal injury may be caused by allopurinol side effects. Compared with a model control group, the serum uric acid content of mice in each dose group of low, medium and high tuna oligopeptide powder is obviously reduced, and the statistical difference (P < 0.01) is provided, so that the tuna oligopeptide is prompted to effectively reduce the uric acid level; adenosine deaminase in the serum of the low, medium and high dose groups of tuna oligopeptide powder has a reduction trend and is in a dose relationship, but has no statistical difference; the tuna oligopeptide powder has no significant influence on the contents of creatinine, urea, blood sugar, total cholesterol, triglyceride, alanine aminotransferase and aspartate aminotransferase in serum.
TABLE 2 Biochemical index comparison of blood of mice in each group: (
）
（*P＜0.05；**P＜0.01）
(iii) Effect of Xanthine Oxidase (XOD) in mouse serum
TABLE 3 Oxidation of xanthine in the serum of groups of miceComparison of enzyme (XOD) content: (
）
（*P＜0.05；**P＜0.01）
The content of Xanthine Oxidase (XOD) in the serum of the mice is detected by an Elisa method. The results showed a significant increase in the levels of xanthine oxidase in the sera of the model group mice compared to the blank control group (. about.p < 0.01). Compared with the model group, the content of xanthine oxidase in the serum of mice in the positive control group, the tuna oligopeptide powder is obviously reduced (P < 0.01) in the low, medium and high dose groups, and the dose groups of the test article are in dose relation.
(iv) Effect of Adenosine Deaminase (ADA) and Xanthine Oxidase (XOD) content in liver tissue of mice
The content of Adenosine Deaminase (ADA) in the livers of each group of mice was determined using a blood biochemical analyzer. The results showed that the liver adenosine deaminase content in the model mice was significantly increased (. about.P. < 0.05) compared to the blank control group. Compared with the model group, the content of adenosine deaminase in the liver of the positive control group mouse is obviously reduced (P is less than 0.05), and the content of adenosine deaminase in the liver of the tuna oligopeptide powder of each dosage group mouse is not obviously changed.
The content of Xanthine Oxidase (XOD) in the livers of each group of mice was determined by the Elisa method. The results showed a significant increase in the level of xanthine oxidase in the liver of the model mice compared to the blank control group (. about.P < 0.01). Compared with the model group, the content of the xanthine oxidase in the liver of the positive control group mice is obviously reduced (P is less than 0.01), and the content of the xanthine oxidase in the liver of the tuna oligopeptide powder is obviously reduced (P is less than 0.01) in the low, medium and high dose groups of mice, and is in dose relation.
TABLE 4 comparison of Adenosine Deaminase (ADA) and Xanthine Oxidase (XOD) content in liver tissue of mice: (
）
（*P＜0.05；**P＜0.01）
(v) Effect on liver XOD and ADA m RNA expression
TABLE 5 liver XOD and ADA m RNA expression in various groups of mice: (
）
（*P＜0.05；**P＜0.01）
Compared with a blank control group, the expression of the liver XOD and ADA of the mice in the model group is obviously increased (P is less than 0.01), compared with the model control group, the expression of the positive control group XOD and ADA is obviously reduced (P is less than 0.01), the expression of the low-dose group ADA of the tuna oligopeptide powder is obviously reduced (P is less than 0.05), the expression of the XOD and ADA in the medium-dose group is obviously reduced (P is less than 0.01), and the expression of the XOD in the high-dose group is obviously reduced (P is less than 0.01).
The experimental results are summarized as follows: the positive control group showed a significant decrease in body weight (P < 0.01) compared to the model control group. Compared with the model control group, the positive control group has obviously increased creatinine and urea levels (P < 0.01), which indicates that allopurinol has damage to renal function and aggravates the renal metabolism burden, while the low, middle and high groups of the tuna have creatinine and urea values which are almost the same as those of the model group and the blank group, do not influence the urinary metabolism and aggravate the renal burden. The serum uric acid content of the model group is obviously increased compared with that of the blank control group, the serum uric acid and xanthine oxidase contents of the tuna oligopeptide powder in the low, medium and high dose groups are obviously reduced (P < 0.01), the dose relationship is shown, the statistical difference is realized, the xanthine oxidase content in liver tissues is obviously reduced (P < 0.01), and the dose relationship is also shown, and the statistical difference is realized. Compared with the model control group, the expression of ADA in the low-dose tuna peptide group is obviously reduced (P is less than 0.05), the expression of XOD and ADA in the high-dose tuna peptide group is obviously reduced (P is less than 0.01), and the expression of XOD in the low-dose tuna peptide group is obviously reduced (P is less than 0.01). Therefore, the research preliminarily verifies that the tuna oligopeptide powder can inhibit ADA/XOD activity and mRNA expression in mouse serum, promotes the metabolism of uric acid in a mouse body, and reduces the high uric acid level of the mouse.
Example 4
Tuna oligopeptide powder composition product human body trial feedback
The tuna oligopeptide powder composition products obtained in examples 1 and 2 are provided for patients with hyperuricemia to take test clothes, and the test clothes are required to determine blood uric acid value indexes before and after taking test clothes, and detailed data are shown in the following table 6.
TABLE 6 tuna oligopeptide powder composition product human trial feedback
Therefore, after a trial man eats the formula product for one period, the blood uric acid value is obviously reduced and is close to the upper limit of the normal uric acid value of a male by 420 mu mol/L, the gout condition is effectively relieved, and the expected target of the formula product is realized.
The diseases such as hyperuricemia, chronic renal function injury and the like are treated mainly by medicaments at the present stage, so that toxic and side effects with different degrees are generated, all raw and auxiliary materials and finished products in the formula are qualified for food production through audit certification, and the food has no toxic and side effects on a human body after being eaten for a long time and has lower cost than the medicaments. The formula product can effectively adjust the taste of the formula by adding various fruit juice powder, sour sweetener and other ingredients, is sour, sweet, tasty and pleasant, and is easy to accept by consumers. The mixed powder is prepared into strip-shaped instant food by a food processing technical means, is easy to store and carry, can be eaten after being brewed with warm water, and is convenient and practical. Although the food formula is not as fast as the drug in effect taking speed, the uric acid level can be reduced by consistent taking, the pain of patients with hyperuricemia and gout can be effectively relieved, the toxic and side effects of western medicines are avoided, the renal function can be assisted to be improved, the renal burden is lightened, the health level of patients with hyperuricemia can be improved, the social benefit is remarkable, and the general requirements of 'healthy Chinese 2030' planning compendium are met.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (8)
1. The tuna oligopeptide powder composition is characterized by comprising the following components in parts by weight: 25-30 parts of tuna oligopeptide powder, 10-15 parts of fructo-oligosaccharide, 15-20 parts of fish collagen peptide powder, 15-20 parts of acerola juice powder, 15-20 parts of juicy peach juice powder, 5-10 parts of erythritol, 1-2 parts of compound sour sweetener and 3-4 parts of water-soluble sugar; the water-soluble sugar is fucoidin or chitosan oligosaccharide.
2. The tuna oligopeptide powder composition of claim 1, wherein the content of tuna oligopeptide functional dipeptide from 20 to 60 mg.
3. The tuna oligopeptide powder composition of claim 1, wherein the compound sour sweetener comprises DL-malic acid, citric acid, sodium citrate and a sweetener, the weight ratio of DL-malic acid, citric acid, sodium citrate and the sweetener is 3:2:1:1, and the sweetener is sucralose, mogroside or stevioside.
4. The tuna oligopeptide powder composition of claim 1, wherein the powder size of the tuna oligopeptide powder composition is 60-100 mesh.
5. A method of preparing a tuna oligopeptide powder composition according to any one of claims 1 to 4, comprising:
1) preparation of tuna oligopeptide powder:
(a) pretreatment of raw materials: thawing raw water of tuna, taking tuna meat, washing twice with tap water, and washing once with purified water; (b) crushing: pulverizing tuna meat for 30min with an automatic chopper mixer to obtain homogenate; (c) pressure heat treatment: pouring the homogenate into a pressure stirring pot, adding purified water with the mass 2 times that of the fish meat, heating to 120 ℃, preserving heat for 2 hours, closing the pressure stirring pot, and naturally cooling to normal temperature and normal pressure; (d) enzymolysis: transferring the homogenate after the heat treatment into an enzymolysis tank for enzymolysis to obtain an enzymolysis liquid; (c) centrifuging: heating the enzymolysis solution to 60 ℃, filtering the enzymolysis solution by a 100-mesh filter screen, standing for 2 hours, then feeding the enzymolysis solution into a centrifugal machine, centrifuging at 6900r/min, feeding at a speed of 0.5t/h, discharging slag for 10 min/time, and feeding the centrifuged feed liquid into a liquid storage tank; (d) and (3) decoloring: heating the feed liquid to 50 ℃, adding active carbon accounting for 5% of the fish meat mass, and stirring for 30 min; (e) and (3) filtering: filter pressing the feed liquid for 2 times by using diatomite as a filter cake; (f) concentration and purification: pumping the filtrate into a liquid storage tank, and circulating through an ultrafiltration device, wherein the cut-off molecular weight of the ultrafiltration membrane is 10000Da, and the pressure is lower than 0.2 MPa; nano-filtering, concentrating and desalting to remove 80% of water; transferring the feed liquid into a single-effect evaporator, concentrating for the second time until the concentration of the feed liquid is 12-16%, controlling the pressure to be 0.08MPa, discharging evaporation condensate water every half hour, and heating to 80 ℃; (g) spray drying: preheating spray drying equipment, heating the feed liquid to 90 ℃, pumping the feed liquid into a spray drying tower, controlling the temperature at 150 ℃, the feeding amount at 50kg/h and the powder collecting frequency at 1 h/time; (h) and (3) packaging and storing: putting the dried powder into a food-grade plastic bag, fastening the opening of the bag, labeling, and storing in a warehouse;
2) preparing fish collagen peptide powder:
(a) pretreatment of raw materials: cleaning Alaska deepened sea cod skin twice with tap water, and cleaning with purified water once; (b) crushing: crushing the cod skin for 10min by using an automatic cutting and mixing machine to obtain homogenate; (c) pressure heat treatment: pouring the homogenate into a pressure stirring pot, adding purified water 2 times the weight of the fish skin, heating to 120 ℃, keeping the temperature for 30min, closing the pressure stirring pot, and naturally cooling to normal temperature and normal pressure; (d) enzymolysis: transferring the homogenate after the heat treatment into an enzymolysis tank for enzymolysis to obtain an enzymolysis liquid; (e) centrifuging: heating the enzymolysis solution to 60 ℃, filtering the enzymolysis solution by a 100-mesh filter screen, standing for 2 hours, then feeding the enzymolysis solution into a centrifugal machine, centrifuging at 6900r/min, feeding at a speed of 0.5t/h, discharging slag for 10 min/time, and feeding the centrifuged feed liquid into a liquid storage tank; (f) and (3) decoloring: heating the feed liquid to 50 ℃, adding active carbon accounting for 5% of the weight of the fish skin, and stirring for 30 min; (g) and (3) filtering: filter pressing the feed liquid for 2 times by using diatomite as a filter cake; (h) concentration and purification: pumping the filtrate into a liquid storage tank, circulating through an ultrafiltration device, intercepting the molecular weight of the ultrafiltration membrane to 10000Da and concentrating to remove 80% of water, wherein the pressure is lower than 0.2 MPa; transferring the feed liquid into a single-effect evaporator, concentrating for the second time until the concentration of the feed liquid is 12-16%, controlling the pressure to be 0.08MPa, discharging evaporation condensate water every half hour, and heating to 80 ℃; (i) spray drying: preheating spray drying equipment, heating the feed liquid to 90 ℃, pumping the feed liquid into a spray drying tower, controlling the temperature at 150 ℃, the feeding amount at 50kg/h and the powder collecting frequency at 1 h/time; (j) and (3) packaging and storing: putting the dried powder into a food-grade plastic bag, fastening the opening of the bag, labeling, and storing in a warehouse;
3) tuna oligopeptide powder composition:
the tuna oligopeptide powder composition comprises the following components in parts by weight: 25-30 parts of tuna oligopeptide powder, 10-15 parts of fructo-oligosaccharide, 15-20 parts of fish collagen peptide powder, 15-20 parts of acerola juice powder, 15-20 parts of juicy peach juice powder, 5-10 parts of erythritol, 1-2 parts of compound sour sweetener and 3-4 parts of water-soluble sugar; the water-soluble sugar is fucoidin or chitosan oligosaccharide;
weighing the raw materials in proportion, sequentially sieving with 60 mesh sieve, transferring the sieved raw materials into a V-shaped mixer, starting a power supply to automatically mix for 35min, transferring the uniformly mixed powder to a full-automatic quantitative packaging machine, adjusting the metering value to 4 g/bag, and automatically packaging.
6. The method for preparing the tuna oligopeptide powder composition according to claim 5, wherein the enzymolysis process in the preparation of the tuna oligopeptide powder is as follows: transferring the heat-treated homogenate into an enzymolysis tank, adding purified water with the mass 8 times that of fish, stirring uniformly, adjusting the temperature to 50-52 ℃, adjusting the pH to =9, adding alkaline protease with the mass 0.5% of fish for enzymolysis for 2h, adjusting the pH of the homogenate to 7, sequentially adding neutral protease, compound protease and flavor enzyme with the mass 0.5% of fish, performing enzymolysis for 1h, controlling the temperature to 50-52 ℃ in the enzymolysis process, and stirring once every 10min to obtain an enzymolysis solution.
7. The method for preparing the tuna oligopeptide powder composition according to claim 5, wherein the enzymolysis process in the preparation of the tuna collagen peptide powder is as follows: transferring the heat-treated homogenate into an enzymolysis tank, adding purified water 6 times the mass of the fish skin, uniformly stirring, adjusting the temperature to 53-55 ℃, adjusting the pH to =9, adding alkaline protease 0.5% of the mass of the fish skin for enzymolysis for 2h, adjusting the pH of the homogenate to 7, sequentially adding neutral protease 0.5% of the mass of the fish skin and flavor enzyme, performing enzymolysis for 1h, controlling the temperature to 53-55 ℃ in the enzymolysis process, and stirring once every 10 min.
8. Use of a tuna oligopeptide powder composition according to any one of claims 1 to 4 for reducing serum uric acid levels.
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| Publication number | Priority date | Publication date | Assignee | Title |
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Application publication date: 20201023 |