CN116172191A - Nutritional composition and its application in preparing anti-inflammatory products - Google Patents
Nutritional composition and its application in preparing anti-inflammatory products Download PDFInfo
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Abstract
The present invention provides nutritional compositions and their use in the preparation of anti-inflammatory products. The present invention provides the use of a nutritional composition in the preparation of an anti-inflammatory product; the nutritional composition comprises protein, fat, carbohydrate and calcium salt; the proteins include lactoferrin and osteopontin; in the nutritional composition, the total protein content is (9-50 g)/100 g, the fat content is (12-30 g)/100 g, the carbohydrate content is (30-60 g)/100 g, the lactoferrin content is (0.005-3 g)/100 g, the osteopontin content is (0.01-1 g)/100 g, and the calcium content is (0.1-2 g)/100 g. The nutritional composition contains lactoferrin, osteopontin and calcium, has a synergistic inhibition effect on chronic inflammation which is easy to generate for tumor patients, wound stress patients and the like, has high safety, and is worthy of popularization and application.
Description
Technical Field
The present invention relates to the field of food technology, and more specifically to nutritional compositions and their use in the preparation of anti-inflammatory products.
Background
Tumors are the most common disease which endangers life and health of people at present, and the mortality rate is the first. Along with the continuous development of medical technology, the tumor treatment technology is also continuously improved and promoted, more and more tumor patients are improved in disease condition, but the number of recurrence and metastasis cases is still more, and related medical staff are required to attach importance to rehabilitation and health care of the tumor patients. Several epidemiological studies have shown that more than 2/3 of tumor patients can be prevented by taking appropriate lifestyles. It was found that a lactoferrin enriched diet can act as a prophylactic. Osteopontin is an important component of the extracellular matrix of tumors and is considered a key marker for the progression of many tumors.
Patients with post-traumatic stress disorder may develop emotional symptoms, including increased alertness, sensitivity, irritability to the environment, avoidance and numbness to daily life, or become turnera. Surgical trauma stress can induce a peripheral inflammatory response in the body, which destroys the integrity of the blood brain barrier by inflammatory cells, pro-inflammatory factors, or activates a series of neural and humoral mechanisms such as vagal primary neurons by transport proteins, related receptors, or by immune related stimulation, thereby activating microglial cells, producing a series of inflammatory mediators leading to an inflammatory response in the central nervous system, and eventually damaging the hippocampal brain region leading to PND. In the implementation of nutritional support for trauma patients, it is important to provide sufficient and appropriate proportions of heat to the patient, and the heat balance is closely related to mortality in critically ill patients, and damage to the intestinal mucosal barrier can occur early in the trauma.
Lactoferrin is involved in a wide range of biological functions including anti-infective, anti-inflammatory and reducing the detrimental host response to acute enteritis. Among them, lactoferrin plays an important immunological role as a key immunonutrient in breast milk. In piglets fed lactoferrin, immune cells produce a strong pro-inflammatory response when the bacteria trigger. These studies demonstrate the benefits of dietary lactoferrin in preventing infection, late sepsis and necrotizing enterocolitis.
The osteopontin is a multifunctional protein with biological activity, exists in most tissues and body fluids, and is found to be capable of remarkably promoting human small intestine gland fossa cell proliferation, reducing the activity of intestinal neutrophil in acute enteritis mice, reducing the secretion of a series of pro-inflammatory factors such as tumor necrosis factor (TNF-alpha), interferon gamma and the like, and reducing harmful host reactions by enhancing and restoring epithelial barriers and the like.
In the prior art, the anti-inflammatory effect of the calcium ion, lactoferrin and osteopontin, which is easily generated in adult disease states of tumor patients, trauma stress patients and the like, is not clear, and needs to be further explored.
Disclosure of Invention
In view of the above, the present invention aims to provide a nutritional composition and an application thereof in preparing anti-inflammatory products, wherein the nutritional composition contains lactoferrin, osteopontin and calcium, has a synergistic inhibition effect on chronic inflammation which is easy to occur in adult diseased states of tumor patients, trauma stress patients and the like, has high safety, and is worthy of popularization and application.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
use of a nutritional composition for the preparation of an anti-inflammatory product;
the nutritional composition comprises protein, fat, carbohydrate and calcium salt;
the proteins include lactoferrin and osteopontin;
in the nutritional composition, the total protein content is (9-50 g)/100 g, the fat content is (12-30 g)/100 g, the carbohydrate content is (30-60 g)/100 g, the lactoferrin content is (0.005-3 g)/100 g, the osteopontin content is (0.01-1 g)/100 g, and the calcium content is (0.1-2 g)/100 g.
In the invention, the mass ratio of the lactoferrin to the osteopontin to the calcium is 1 (0.1-5) (0.001-40); the mass ratio of the lactoferrin to the osteopontin to the calcium is preferably 1 (0.1-5): (0.011 to 0.06), more preferably 1: (0.1-5): (0.011-0.02).
In the present invention, the anti-inflammatory includes inhibiting the expression of a pro-inflammatory factor-related gene, and/or down-regulating the expression of an inflammatory pathway-related protein;
the pro-inflammatory factors include IL-1β and/or IL-6, preferably IL-1β;
the inflammatory pathway related proteins include one or more of TLR4, p-p65, p-STAT 3.
In the present invention, the anti-inflammatory product is preferably a food product;
the anti-inflammatory product is a formula food with special medical application; special medical use formulas, which are formulas specially processed and formulated to meet the special needs of people with limited eating, digestive absorption disorder, metabolic disorder or specific disease states for nutrients or diet, include special medical use infant formulas applicable to 0 to 12 months of age and special medical use formulas applicable to people over 1 year of age;
the anti-inflammatory is preferably against chronic inflammation;
the patient producing the chronic inflammation is preferably a tumor patient or a trauma stress patient. Chronic inflammation (chronic inflammation) is a long-term, slowly progressing inflammation that can last for several months to years, and sometimes is not known by itself under the duration of chronic inflammation. Chronic inflammation and related diseases are the leading causes of death in humans. Stress can cause chronic systemic low-grade inflammation of organisms, wherein the chronic low-grade inflammation is soil with various diseases, and further has negative effects on human health; tumor development and progression are associated with chronic inflammation. The microenvironment consisting of chronic inflammation is critical for the occurrence, development and metastasis of tumors, and is also the response of the body to tumors, and in general, the inflammation has a promoting effect on the tumors.
In the nutritional composition of the present invention, the total protein content is (15-35 g)/100 g, the fat content is (12-20 g)/100 g, the carbohydrate content is (32-50 g)/100 g, the lactoferrin content is (0.005-0.8 g)/100 g, the osteopontin content is (0.03-0.5 g)/100 g, and the calcium content is (0.1-1 g)/100 g.
For one embodiment of the invention, the nutritional composition has a total protein content of 34g/100g, a fat content of 12g/100g, a carbohydrate content of 32g/100g, a lactoferrin content of 600mg/100g, a osteopontin content of 60mg/100g, and a calcium content of 300mg/100g.
For one embodiment of the invention, the nutritional composition has a total protein content of 33g/100g, a fat content of 18.5g/100g, a carbohydrate content of 34g/100g, a lactoferrin content of 16mg/100g, a osteopontin content of 80mg/100g, and a calcium content of 640mg/100g.
In the present invention, the lactoferrin is derived from one or more of cow milk or a product thereof, sheep milk or a product thereof, camel milk or a product thereof, preferably whey protein powder.
In the present invention, the osteopontin is derived from one or more of cow milk or a product thereof, sheep milk or a product thereof, camel milk or a product thereof, preferably whey protein powder.
In the present invention, the calcium salt is derived from one or more of calcium carbonate, calcium gluconate, calcium citrate, calcium lactate, calcium hydrogen phosphate, calcium L-threonine, calcium glycinate, calcium aspartate, calcium acetate, calcium chloride, tricalcium phosphate, vitamin E calcium succinate, calcium glycerophosphate, calcium sulfate, calcium dihydrogen phosphate, milk mineral salts, calcium caseinate, calcium malate, and calcium ascorbate. Milk mineral salt is also called milk calcium, whey mineral concentrate and whey calcium, and is milk white powder obtained by processing milk through membrane permeation, separation, concentration, spray drying and other processes, and the main component of the milk mineral salt is calcium phosphate, and the milk mineral salt also comprises rich nutritional components such as protein, lactose, zinc, phosphorus, sodium, potassium, magnesium and the like, wherein the calcium content is about 23% -28%, the calcium-phosphorus ratio is 2:1, and the milk mineral salt is more beneficial to absorption and utilization of human bodies.
In the invention, the raw materials for providing the protein comprise one or more of sodium caseinate, calcium caseinate, casein, hydrolyzed whey protein, hydrolyzed casein, hydrolyzed milk protein, hydrolyzed milk fat globule membrane protein and separated whey protein;
the raw materials for providing the fat comprise one or more of milk fat, vegetable oil, fish oil and triglyceride; the triglycerides are preferably medium chain triglycerides;
the vegetable oil is one or more selected from sunflower seed oil, corn oil, coconut oil, canola oil, linseed oil, peanut oil, soybean oil, palm oil and walnut oil; preferably sunflower seed oil, corn oil and soybean oil; the addition of vegetable oil provides on the one hand a fat component for the nutritional composition and on the other hand linoleic acid and also alpha-linolenic acid; more preferably, the raw material thereof contains 0 to 150 parts by weight of sunflower seed oil based on 1000 parts by weight of the nutritional composition; corn oil 0-80 weight portions; 0-80 parts by weight of soybean oil; the addition of fish oil can not only provide a fat component, but also unsaturated fatty acids; unsaturated fatty acids include docosahexaenoic acid (DHA), arachidonic acid (ARA), and eicosapentaenoic acid (EPA);
providing the carbohydrate as a raw material comprising one or more of lactose, gelatinized starch, maltodextrin, solid corn syrup, glucose syrup, and dietary fiber; the carbohydrates in the nutritional compositions of the present invention are derived in part from lactose and in part from non-lactose sources including, but not limited to, gelatinized starch, maltodextrin, solid corn syrup, glucose syrup; that is, in the nutritional composition of the present invention, the carbohydrate-providing raw material includes raw material lactose and starch-based material pre-hydrolyzed and gelatinized in addition to the lactose-containing base raw material; preferably, the nutritional composition comprises, based on 1000 parts by weight, the following raw materials: lactose 0-580 weight portions and non-lactose material 0-580 weight portions; the specific addition amount of lactose of the above raw materials may be adjusted within the range so that the carbohydrate content in the nutritional composition of the present invention is 50 to 58g/100g; the dietary fiber is preferably fructooligosaccharides; fructo-oligosaccharides (FOS) are a natural active substance with sweetness 0.3-0.6 times that of sucrose. The fructo-oligosaccharide not only maintains the pure sweet property of the sucrose, but also is more refreshing than the sweet taste of the sucrose. Has health promoting effects such as regulating intestinal flora, proliferating Bacillus bifidus, promoting calcium absorption, regulating blood lipid, regulating immunity, and resisting dental caries;
the nutritional composition further comprises one or more of amino acids, vitamins;
the amino acids include arginine and/or tryptophan;
in one embodiment of the present invention, the nutritional composition comprises the following raw materials based on 1000 parts by weight: 8-50 parts by weight of DHA and 14-28 parts by weight of ARA; EPA is 10-140 parts by weight; arginine is 0 to 10 weight parts; tryptophan is 0 to 10 weight portions; 0 to 7.5 parts by weight of lactoferrin; 0-75 parts by weight of osteopontin; 7-50 parts by weight of compound nutrient bag containing calcium powder, vitamins and minerals.
In the nutritional composition, the compound nutrient package is a combination of nutritional ingredients meeting the national standard, and different addition amounts are used according to different formulas. The nutritional composition of the invention can optionally adopt any one or any combination of the following compound nutritional package components if nutrients are added according to needs. Preferably, the compound nutrient package at least comprises compound vitamin, calcium powder and mineral nutrient packages, and the content of each component in the compound nutrient package is as follows:
1) A compound vitamin nutrition package, wherein each gram of compound vitamin nutrition package comprises:
taurine: 140-340 mg
Vitamin a: 1700-5800 mu gRE
Vitamin D: 25-70 mu g
Vitamin B 1 ：2000～6800μg
Vitamin B 2 ：3000～6900μg
Vitamin B 6 ：1700～4000μg
Vitamin B 12 ：8～20μg
Vitamin K 1 ：200～700μg
Vitamin C: 0-700 mg
Vitamin E: 10-70 mg alpha-TE
Nicotinamide: 10000-41550 mug
Folic acid: 350-920 mu g
Biotin: 70-245 mug
Pantothenic acid: 7100-25230 mu g
Inositol: 0-250mg
L-carnitine: 0-60mg
2) Mineral one nutrition package, in every gram of mineral one nutrition package:
iron: 20-110 mg
Zinc: 23-90 mg
Copper: 2000-4180 mug
Iodine: 500-995 mug
Selenium: 0-200 mu g
Manganese: 0-579 mug
3) Mineral two nutrition package, in every gram mineral two nutrition package:
sodium: 40-100 mg
Potassium: 200-500 mg
4) Mineral triad package, per gram of mineral triad package:
calcium: 200-500 mg
Phosphorus: 75-300 mg
5) A compound magnesium chloride nutrition package, wherein each gram of compound magnesium chloride nutrition package comprises:
magnesium: 80-170 mg
6) Choline chloride nutrition packet, per gram of choline chloride nutrition packet:
choline: 300-950 mg
The base material of the compound nutrient packet is preferably lactose, maltodextrin, solid corn syrup or L-sodium ascorbate. Based on 1000 parts by weight of the nutritional composition, the addition amount of the compound nutrient package is 7-52 parts by weight, wherein the compound vitamin nutrient package is preferably 2-4 parts by weight, the mineral secondary nutrient package is preferably 2-20 parts by weight, the mineral tertiary nutrient package is preferably 0.5-20 parts by weight, the mineral primary nutrient package is preferably 0.5-6 parts by weight, the magnesium chloride is 0-7 parts by weight, the choline chloride is 0-3 parts by weight, and the base material of each nutrient package is preferably maltodextrin, lactose or sodium L-ascorbate.
The compound materials used to provide each nutrient in the nutritional package may have interactions. For example, sulfate can accelerate the oxidative destruction process of vitamins, which can reduce their utilization. Because sulfate occurs in ionic form in aqueous solution, it acts as an oxidizing agent to induce oxidation of vitamins during the oxidation reaction, thereby destroying the vitamin structure. The trace elements have different capacities in oxidation-reduction reaction, and the activities of copper, zinc and iron are strongest, and manganese and selenium are used for times. Vitamin B and vitamin C are susceptible to copper ions, vitamin B 2 Is susceptible to iron ions.
To ensure nutrient utilization efficiency, the present invention selects stable nutrient dosage forms, such as: the retinol is selected from retinol acetate, and retinol contains 1 hydroxyl group and 5 double bonds, so that the retinol is very easy to oxidize, but the stability of the retinol is greatly improved under the form of the retinol acetate; vitamin E is selected from tocopheryl acetate, the tocopherols are unstable, but the stability of tocopheryl acetate is higher; vitamin B 1 Selecting thiamine nitrate, wherein the thiamine nitrate is more stable than thiamine hydrochloride; the vitamin C is L-sodium ascorbate.
The above-mentioned compound nutrient package component contents refer to the nutrient component contents in other raw materials of the nutrient composition excluding the additive amount for enhancing the nutrient substances, and according to a preferred embodiment of the present invention, the formula of the nutrient composition of the present invention includes:
it will be appreciated that the specific amounts of the ingredients in the nutritional compositions of the present invention should be determined by adjusting the amounts of the ingredients to meet the product specifications of the nutritional composition.
In the nutritional composition of the invention, the raw materials are commercially available, and the raw materials are selected to meet the requirements of relevant standards, wherein the lactoferrin, the osteopontin and the calcium salt meet the requirements of the invention. In addition, the compound nutrient package can also be self-compounded. The invention adopts 'compound' for convenience of expression, and does not mean that all components in the compound are mixed together and then applied. All raw materials should be added and used on the premise of meeting related regulations.
The present invention also provides nutritional compositions including proteins, fats, carbohydrates, and calcium salts;
the proteins include lactoferrin and osteopontin;
in the nutritional composition, the total protein content is (9-50 g)/100 g, the fat content is (12-30 g)/100 g, the carbohydrate content is (30-60 g)/100 g, the lactoferrin content is (0.005-3 g)/100 g, the osteopontin content is (0.01-1 g)/100 g, and the calcium content is (0.1-2 g)/100 g.
In the present invention, the nutritional composition specifically comprises 0 to 200 parts by weight of whey protein powder; 0-100 parts by weight of sodium caseinate; 0-100 parts by weight of casein; 0-400 parts by weight of solid corn syrup; 0-400 parts by weight of maltodextrin; 0-50 parts by weight of resistant dextrin; 0-50 parts by weight of fructo-oligosaccharide; 0-150 parts by weight of sunflower seed oil; corn oil 0-40 weight portions; 20-80 parts by weight of soybean oil; 0-100 parts by weight of medium chain triglyceride; 0-10 parts by weight of tryptophan; arginine 0-10 weight portions; 0.01 to 7.5 parts by weight of lactoferrin; 0.75 to 37.5 parts by weight of osteopontin; 0.0075-300 parts by weight of calcium salt; 7-50 parts by weight of compound nutrient packet containing calcium powder, vitamins and minerals; DHA 5-80 weight portions; 0-100 parts by weight of glutamine; EPA 10-140 weight portions.
The method for preparing the nutritional composition is not particularly limited, and conventional methods in the art may be employed. The preparation method of the nutritional composition comprises the following steps: proportioning, homogenizing, concentrating, sterilizing, spray drying and dry mixing to obtain a finished product;
the preparation method preferably comprises the following steps:
1) Powder adding: the powder raw materials are uniformly added into a powder preparation tank for storage through an air-assisted system after being metered according to the formula;
2) Vacuum powder suction: various powder raw materials in the powder preparation tank are sucked into the vacuum mixing tank through the vacuum system;
3) Melting and oil preparing: placing the grease specified in the formula into a grease melting room according to the formula requirement;
4) And (3) storing the mixed oil: the mixed oil is stored in an oil storage tank in a heat-preserving way;
5) Weighing: pumping the mixed oil into a mixing tank through an oil pump according to the formula requirement;
6) Nutrient dissolution and addition: dissolving the compound nutrient package with water, and adding into a mixing tank;
7) Lactoferrin, osteopontin and calcium salt solubilization addition: dissolving osteopontin and calcium salt according to the step in the step 6, and adding the dissolved osteopontin and calcium salt into a mixing tank;
8) And (3) filtering: filtering the mixed feed liquid through a filter screen to remove physical impurities in the raw materials;
9) Homogenizing: homogenizing the filtered feed liquid by a homogenizer;
10 Cooling and storing: the homogenized feed liquid enters a plate heat exchanger for cooling;
11 Concentration sterilization: double-effect concentration is used during production; the discharge concentration is 48% -52% of dry matter;
12 Concentrated milk storage, pre-heating filtration, spray drying: temporarily storing the concentrated milk in a concentrated milk balance tank; preheating, filtering and drying the materials, and agglomerating the obtained fine powder at the top of the tower;
13 Fluidized bed drying and cooling: the powder from the drying tower is subjected to fluidized bed (first-stage) secondary drying and then is cooled by the fluidized bed (second-stage);
14 Split charging: weighing DHA, ARA and lactoferrin by powder workshop personnel according to the formula requirement, bagging and subpackaging;
15 Dry blending): mixing the weighed DHA, EPA, ARA, lactoferrin and milk powder uniformly in a dry mixer; if the protein content is higher, the protein can be added by uniformly mixing part of casein, whey protein powder and sodium caseinate in a dry mixing mode;
16 Screening powder: the granularity of the milk powder is uniform through a vibrating screen, and the powder slag is scrapped;
17 Powder discharge: receiving powder by a sterilized powder collecting box, and conveying the powder from a powder outlet room to a powder feeding room;
18 Powder) is added: pouring milk powder into a powder storage tank on a size packaging machine according to the packaging requirement;
19 Packaging: 800 g of automatic packaging machine fills nitrogen for packaging, and the oxygen content is lower than 1% during nitrogen filling; 900 g of iron cans automatically fill nitrogen and pack oxygen content lower than 5%;
20 Boxing: packaging the packaged small bags into a paper box, adding a powder spoon at the same time, and sealing by a box sealing machine;
21 Inspection of the finished product: sampling and checking the packaged product according to a checking plan;
22 Warehousing and storing: the qualified products are stored in warehouse, and the storage is required at normal temperature, and the humidity is less than or equal to 65%.
The nutritional composition provided by the invention contains lactoferrin, osteopontin and calcium, has a synergistic inhibition effect on inflammation, particularly has a synergistic inhibition effect on chronic inflammation which is easy to generate in adult diseased states of tumor patients, trauma stress patients and the like, has high safety, and is worthy of popularization and application. The experimental examples prove that the three components of lactoferrin, osteopontin and calcium can relieve the expression of the RAW264.7 macrophage inhibiting the pro-inflammatory factor at the mRNA level under the induction of LPS under the limit proportion, and can down regulate the expression of the protein related to the inflammatory pathway at the protein level, thereby relieving the influence of inflammatory reaction.
Drawings
FIG. 1 is a flow chart of LF-OPN-calcium ion treatment of macrophages in an experimental example of the invention;
FIG. 2 shows the effect of LF-OPN-calcium combination on cells under LPS in experimental examples of the invention;
FIG. 3 shows the effect of lactoferrin, osteopontin undigested matter on cell activity in experimental examples of the present invention;
FIG. 4 shows the effect of lactoferrin and osteopontin digests on cell activity in experimental examples of the invention;
FIG. 5 is the effect of LF-OPN-calcium combination on IL-1β mRNA level expression in experimental examples of the invention;
FIG. 6 is the effect of LF-OPN-calcium combination on IL-6mRNA level expression in experimental examples of the invention;
FIG. 7 shows the effect of LF-OPN-calcium combinations on inflammatory pathway-related protein expression in experimental examples of the invention;
FIG. 8 is the effect of LF-OPN combination and LF-OPN-calcium combination on IL-1β mRNA level expression in experimental examples of the invention.
Detailed Description
The technical solutions of the present invention will be clearly and completely described in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
In order to further illustrate the present invention, the following examples are provided. The raw materials used in the following examples of the present invention are all commercially available.
The nutritional composition content in the nutritional package of the present invention is as follows:
1) A compound vitamin nutrition package, wherein each gram of compound vitamin nutrition package comprises:
taurine: 140-340 mg
Vitamin a: 1700-5800 mu gRE
Vitamin D: 25-70 mu g
Vitamin B 1 ：2000～6800μg
Vitamin B 2 ：3000～6900μg
Vitamin B 6 ：1700～4000μg
Vitamin B 12 ：8～20μg
Vitamin K 1 ：200～700μg
Vitamin C: 0-700 mg
Vitamin E: 10-70 mg alpha-TE
Nicotinamide: 10000-41550 mug
Folic acid: 350-920 mu g
Biotin: 70-245 mug
Pantothenic acid: 7100-25230 mu g
Inositol: 0-250mg
L-carnitine: 0-60mg.
2) Mineral one nutrition package, in every gram of mineral one nutrition package:
iron: 20-110 mg
Zinc: 23-90 mg
Copper: 2000-4180 mug
Iodine: 500-995 mug
Selenium: 0-200 mu g
Manganese: 0-579 mug
3) Mineral two nutrition package, in every gram mineral two nutrition package:
sodium: 40-100 mg
Potassium: 200-500 mg
4) Mineral triad package, per gram of mineral triad package:
calcium: 200-500 mg
Phosphorus: 75-300 mg.
The calcium salt in the mineral tri-nutrition bag is calcium carbonate, calcium hydrogen phosphate and tricalcium phosphate.
Example 1 contains tumor patient formula (1000 kg prepared):
70kg of casein, 70kg of whey protein powder, 200kg of sodium caseinate, 320kg of maltodextrin, 60 kg of corn oil, 40 kg of soybean oil, 20kg of medium chain triglyceride, 30 kg of fructo-oligosaccharide, 6 kg of lactoferrin, 0.6 kg of osteopontin, 3kg of calcium salt, 1kg of glutamine, 20kg of compound nutrient package, 50 kg of DHA, 80 kg of EPA, 0.5kg of arginine, 0.5kg of tryptophan and the balance of maltodextrin.
Wherein each gram of the compound nutrient package comprises about 6.0 parts by weight of compound vitamin nutrient package, about 2.0 parts by weight of choline chloride nutrient package, about 10 parts by weight of mineral three-nutrient package, 20 parts by weight of mineral two-nutrient package, about 6 parts by weight of mineral one-nutrient package and about 7.0 parts by weight of compound magnesium chloride nutrient package, and the base material of each nutrient package is maltodextrin.
The specific preparation process of the formula powder containing lactoferrin, osteopontin and calcium salt comprises the following steps:
1) Powder adding: the powder base material is metered according to the formula and then uniformly added into a powder preparation tank for storage through an air-assisted system.
2) Vacuum powder suction: various powder raw materials in the powder preparation tank are sucked into the vacuum mixing tank through the vacuum system.
3) Melting and oil preparing: the oil and fat specified in the formula are put into an oil melting room according to the formula requirement, the temperature of the oil melting room is kept at 50-90 ℃, and after the oil is melted, the oil is pumped into a mixed oil storage tank according to the formula proportion requirement through an oil pump and a flowmeter.
4) And (3) storing the mixed oil: the mixed oil is stored in an oil storage tank in a heat preservation way at the temperature of 40-50 ℃ for less than 12 hours to prevent fat oxidation.
5) Weighing: and pumping the mixed oil into a mixing tank through an oil pump according to the formula requirement.
6) Nutrient dissolution and addition: the compound nutrient package comprising calcium powder, mineral substances and vitamins is dissolved by 100-200 kg of purified water respectively, and then is put into a wet mixing tank, and a tank and a pipeline are added after each 100kg of purified water is used for flushing.
7) Lactoferrin, osteopontin and calcium salt solubilization addition: lactoferrin, osteopontin and calcium salt are dissolved according to the step in step 6 and added into a mixing tank.
8) And (3) filtering: the mixed feed liquid is filtered by a filter screen to remove physical impurities possibly carried in the raw materials.
9) Homogenizing: homogenizing the filtered feed liquid by a homogenizer, wherein the primary pressure of homogenization is 105+/-5 bar, and the secondary pressure is 32+/-3 bar, so that fat globules in the feed liquid are dispersed into uniform fat globules.
10 Cooling and storing: the homogenized feed liquid enters a plate heat exchanger for cooling: cooling to below 20 ℃, temporarily storing in a pre-storing cylinder, entering the next working procedure within 6 hours, and starting the stirrer according to the set requirement.
11 Concentration sterilization: during production, double-effect concentration is used, the sterilization temperature is more than or equal to 83 ℃, and the sterilization time is 25 seconds. The discharge concentration is 48% -52% of dry matter.
12 Concentrated milk storage, pre-heating filtration, spray drying: the concentrated milk is temporarily stored in a concentrated milk balance tank. Preheating to 60-70 ℃ by a scraper preheater, filtering the preheated material by a filter with the aperture of 1mm, pumping the material into a drying tower by a high-pressure pump for spray drying, and agglomerating the fine powder on the top of the tower or a fluidized bed according to requirements. Air inlet temperature: 165-180 deg.c, exhaust temperature 75-90 deg.c, high pressure pump pressure 160-210 bar, and negative tower pressure-4 to-2 mbar.
13 Fluidized bed drying and cooling: the powder from the drying tower is dried again by the fluidized bed (first stage) and then cooled to 25-30 ℃ by the fluidized bed (second stage).
14 Split charging: and weighing DHA, ARA and lactoferrin by powder workshop personnel according to the formula requirement, bagging and subpackaging.
15 Dry blending): and mixing the weighed DHA, EPA, ARA, lactoferrin and milk powder uniformly in a dry mixer.
16 Screening powder: the granularity of the milk powder is uniform through the vibrating screen, and the powder slag is scrapped.
17 Powder discharge: and (3) receiving powder by using a sterilized powder collecting box, and conveying the powder from a powder outlet room to a powder feeding room.
18 Powder) is added: and pouring the milk powder into a powder storage tank on a size packaging machine according to the packaging requirement.
19 Packaging: 800 g automatic packaging machine fills nitrogen and packages, and the oxygen content is lower than 1% during nitrogen filling.
20 Boxing: packaging the packaged small bags into a paper box, adding a powder spoon at the same time, and sealing by a box sealing machine.
21 Inspection of the finished product: sampling and checking the packaged product according to a checking plan.
22 Warehousing and storing: the qualified products are stored in warehouse, and the storage is required at normal temperature, and the humidity is less than or equal to 65%.
In the product of this example, the total protein content was 34g/100g, the fat content was 12g/100g, the carbohydrate content was 32g/100g, the lactoferrin was 600mg/100g, the osteopontin was 60mg/100g, the calcium content was 300mg/100g, and the mass ratio of lactoferrin, osteopontin and calcium was 1:0.1:0.5.
example 2 wound stress patient formula (1000 kg prepared):
60 kg of sodium caseinate, 70kg of casein, 200kg of whey protein powder, 340 kg of maltodextrin, 120 kg of corn oil, 30 kg of soybean oil, 35 kg of medium chain triglyceride, 30 kg of fructo-oligosaccharide, 0.16 kg of lactoferrin, 0.8 kg of osteopontin, 6 kg of calcium salt, 1kg of glutamine, 25 kg of compound nutrient package, 30 kg of DHA, 60 kg of EPA, 1kg of arginine, 1kg of tryptophan and the balance of maltodextrin.
Wherein each gram of the compound nutrient package comprises about 6.0 parts by weight of compound vitamin nutrient package, about 2.0 parts by weight of choline chloride nutrient package, about 20 parts by weight of mineral three-nutrient package, 20 parts by weight of mineral two-nutrient package, about 6 parts by weight of mineral one-nutrient package and about 7.0 parts by weight of compound magnesium chloride nutrient package, and the base material of each nutrient package is maltodextrin.
The product preparation process is as in example 1.
In the product of this example, the total protein content was 33g/100g, the fat content was 18.5g/100g, the carbohydrate content was 34g/100g, the lactoferrin was 16mg/100g, the osteopontin was 80mg/100g, the calcium content was 640mg/100g, and the mass ratio of lactoferrin, osteopontin and calcium was 1:5:40.
experimental example
1. Materials and reagents
RAW264.7 macrophage, calcium chloride, LPS, lactoferrin, osteopontin, PBS buffer solution, porcine trypsin, porcine pepsin and DMEM medium (Gibco, 11995065), wherein the purity of the calcium chloride is more than or equal to 99%, the lactoferrin is from Hilmar1000, and the osteopontin is from
OPN-10。
2. In vitro digestion of LF-OPN-calcium
(1) An experimental group is set, and the raw materials comprise the following components in proportion:
a first group: consists of lactoferrin, osteopontin and calcium chloride, wherein the mass ratio of calcium in the lactoferrin, the osteopontin and the calcium chloride is 1:5:0.06;
second group: consists of lactoferrin, osteopontin and calcium chloride, wherein the mass ratio of calcium in the lactoferrin, the osteopontin and the calcium chloride is 1:1:0.02;
third group: consists of lactoferrin, osteopontin and calcium chloride, wherein the mass ratio of calcium in the lactoferrin, the osteopontin and the calcium chloride is 1:0.15:0.0115;
fourth group: consists of lactoferrin, osteopontin and calcium chloride, wherein the mass ratio of calcium in the lactoferrin, the osteopontin and the calcium chloride is 1:0.1:0.011;
fifth group: consists of lactoferrin, osteopontin and calcium chloride, wherein the mass ratio of calcium in the lactoferrin, the osteopontin and the calcium chloride is 1:3:0.04.
sixth group: consists of lactoferrin, osteopontin and calcium chloride, wherein the mass ratio of calcium in the lactoferrin, the osteopontin and the calcium chloride is 1:0.02:0.012.
seventh group: consists of lactoferrin and osteopontin, wherein the mass ratio of the lactoferrin to the osteopontin is 1:0.2.
(2) LF, OPN and calcium are dissolved in the in-vitro digestive system simulating gastric digestive juice and intestinal digestive juice according to the proportion in the step (1), and digested samples are sterilized by a 0.22 mu m filter membrane before acting on cells.
3. LF-OPN-calcium ion treated macrophages
Will be 2X 10 5 RAW264.7 cells per well were seeded into 6-well plates, 3 replicates per treatment group. The inoculated cells were placed in 5% CO at 37 ℃C 2 After 24h, the DMEM medium without serum and antibiotics containing LF and OPN, calcium ions (monomer or combination after in vitro digestion) was replaced for 6h. LPS was then added to the medium to a concentration of 1. Mu.g/ml for a treatment period of 2h to collect cellsRNA was taken and acted for 6h to collect cell extract proteins, and the test procedure is shown in FIG. 1.
4. Detection index
(1) Cell morphology
RAW264.7 cells were inoculated into 12-well cell culture plates, and after 24 hours, DMEM medium containing LF, OPN, calcium ions and no serum and antibiotics was changed. After 6h incubation of the protein, LPS was added to a concentration of 1. Mu.g/ml for 6h. And photographing RAW264.7 cells treated at different time points by using an inverted microscope, recording cell morphology, and characterizing the differentiation degree.
The experimental results are shown in fig. 2, and in fig. 2, left: blank control group; in (a): a LPS group; right: lactoferrin-osteopontin-calcium combination + LPS group. After LPS action for 6h, RAW264.7 changes the cell shape from round shape to slender shape when it is not differentiated; however, in the case of the advanced action of the active protein-calcium combination for 6 hours, the morphology of the cells after the action of LPS for 6 hours was somewhat closer to that of the cells when they were not differentiated than when LPS alone was added.
(2) Cell Activity
Cytotoxicity of the active protein against RAW264.7 was determined by CCK8 experiments. Cells were cultured in 96-well plates, treated for 12h with serum-and antibiotic-free DMEM medium containing LF-OPN-calcium digest, and treated for 1h with CCK 8-free DMEM medium containing no serum and antibiotic. Absorbance was measured with a microplate reader at a wavelength of 450 nm.
As shown in FIG. 3 and FIG. 4, when the concentrations of LF and OPN are higher than 5mg/ml, the effect on cell activity is large, and when the concentrations are lower than 1mg/ml, the cell toxicity is small, so that the monomer or mixed concentration of LF and OPN is 1mg/ml in the experiment.
(3) Expression of inflammatory factors
RAW264.7 cells were inoculated into 12-well cell culture plates and after 24h the DMEM medium containing LF-OPN-calcium and free of serum and antibiotics was changed. After 6h LPS was added to a concentration of 1. Mu.g/ml. RNA is collected after 2 hours, and the expression level of inflammatory factors such as IL-6, IL-1 beta and the like in mRNA is quantitatively analyzed by RT-qPCR.
IL-1 beta and IL-6 are the most common pro-inflammatory factors, and experimental results are shown in fig. 5 and 6, and it can be seen that the LF and OPN monomer digests can inhibit the expression of inflammatory factor genes, but the composition has a synergistic effect, and in the mixed digests, when the ratio is LF: OPN: ca=1: (0.1-5): (0.011-0.06) are superior to LF, OPN monomers and have statistical differences, as can be seen from fig. 8, lactoferrin, osteopontin and calcium group (LF: OPN: ca=1:0.2:0.012) have better anti-inflammatory effects and have statistical differences than the experimental group of lactoferrin and osteopontin (LF: opn=1:0.2).
(4) Inflammatory pathway-related protein expression
RAW264.7 cells were inoculated into 12-well cell culture plates, and after 24 hours, DMEM medium containing LF, OPN, calcium ions and no serum and antibiotics was changed. After 6h LPS was added to a concentration of 1. Mu.g/ml. And collecting protein samples after 6 hours, and analyzing the expression of TLR4 and NF- κB channel related proteins by Western blot.
After six hours of treatment of RAW264.7 with LPS, LF and OPN monomeric digests and different ratios of complex digests, expression of p-STAT3 in the membrane proteins TLR4, NF- κb pathway key protein p-p65 (p 65), and downstream JNK/STAT pathway was examined. The results are shown in fig. 7, and it can be seen that LF: OPN: ca=1: (0.1-5): (0.011-0.06), especially LF: OPN: ca=1: (0.1-5): the composition of (0.011-0.02) can inhibit the expression of inflammatory pathway protein better than the LF or OPN monomer.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (12)
1. Use of a nutritional composition for the preparation of an anti-inflammatory product;
the nutritional composition comprises protein, fat, carbohydrate and calcium salt;
the proteins include lactoferrin and osteopontin;
in the nutritional composition, the total protein content is (9-50 g)/100 g, the fat content is (12-30 g)/100 g, the carbohydrate content is (30-60 g)/100 g, the lactoferrin content is (0.005-3 g)/100 g, the osteopontin content is (0.01-1 g)/100 g, and the calcium content is (0.1-2 g)/100 g.
2. The use according to claim 1, wherein the mass ratio of lactoferrin, osteopontin and calcium is 1 (0.1-5): 0.001-40.
3. The use according to claim 1, wherein the anti-inflammatory product is a food product.
4. The use according to claim 3, wherein the anti-inflammatory product is a special medical use formula.
5. The use according to claim 1, wherein the anti-inflammatory is against chronic inflammation.
6. The use according to claim 5, wherein the patient suffering from the chronic inflammation is a tumor patient or a trauma stress patient.
7. The use according to claim 1, characterized in that in the nutritional composition the total protein content is (15-35 g)/100 g, the fat content is (12-20 g)/100 g, the carbohydrate content is (32-50 g)/100 g, the lactoferrin content is (0.005-0.8 g)/100 g, the osteopontin content is (0.03-0.5 g)/100 g, and the calcium content is (0.1-1 g)/100 g.
8. The use according to claim 1, wherein the lactoferrin is derived from one or more of cow's milk or a preparation thereof, sheep's milk or a preparation thereof, camel's milk or a preparation thereof;
the osteopontin is one or more from cow milk or a product thereof, sheep milk or a product thereof, camel milk or a product thereof;
the calcium salt is selected from one or more of calcium carbonate, calcium gluconate, calcium citrate, calcium lactate, calcium hydrogen phosphate, L-threonine calcium, calcium glycine, calcium aspartate, calcium acetate, calcium chloride, tricalcium phosphate, calcium vitamin E succinate, calcium glycerophosphate, calcium sulfate, calcium dihydrogen phosphate, milk mineral salt, calcium caseinate, calcium malate, and calcium ascorbate.
9. The use according to claim 1, wherein the raw materials providing the protein comprise one or more of sodium caseinate, calcium caseinate, casein, hydrolysed whey protein, hydrolysed casein, hydrolysed milk protein, hydrolysed milk fat globule membrane protein, isolated whey protein;
the raw materials for providing the fat comprise one or more of milk fat, vegetable oil, fish oil and triglyceride.
10. The use according to any one of claims 1-9, wherein the nutritional composition further comprises one or more of amino acids, nucleotides, vitamins, probiotics, inorganic salts.
11. Nutritional composition characterized by comprising proteins, fats, carbohydrates and calcium salts;
the proteins include lactoferrin and osteopontin;
in the nutritional composition, the total protein content is (9-50 g)/100 g, the fat content is (12-30 g)/100 g, the carbohydrate content is (30-60 g)/100 g, the lactoferrin content is (0.005-3 g)/100 g, the osteopontin content is (0.01-1 g)/100 g, and the calcium content is (0.1-2 g)/100 g.
12. Nutritional composition according to claim 11, characterized in that it comprises in particular 0-200 parts by weight of whey protein powder; 0-100 parts by weight of sodium caseinate; 0-100 parts by weight of casein; 0-400 parts by weight of solid corn syrup; 0-400 parts by weight of maltodextrin; 0-50 parts by weight of resistant dextrin; 0-50 parts by weight of fructo-oligosaccharide; 0-150 parts by weight of sunflower seed oil; corn oil 0-40 weight portions; 20-80 parts by weight of soybean oil; 0-100 parts by weight of medium chain triglyceride; 0-10 parts by weight of tryptophan; arginine 0-10 weight portions; 0.01 to 7.5 parts by weight of lactoferrin; 0.75 to 37.5 parts by weight of osteopontin; 0.0075-300 parts by weight of calcium salt; 7-50 parts by weight of compound nutrient packet containing calcium powder, vitamins and minerals; DHA 5-80 weight portions; 0-100 parts by weight of glutamine; EPA 10-140 weight portions.
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