CN114451563A - Composition, application thereof and food - Google Patents

Composition, application thereof and food Download PDF

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Publication number
CN114451563A
CN114451563A CN202210180875.9A CN202210180875A CN114451563A CN 114451563 A CN114451563 A CN 114451563A CN 202210180875 A CN202210180875 A CN 202210180875A CN 114451563 A CN114451563 A CN 114451563A
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vitamin
calcium
content
formula
protein
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Inventor
王永丽
段素芳
周名桥
王燕霞
刘彪
王晓彦
司徒文佑
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Inner Mongolia Yili Industrial Group Co Ltd
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Inner Mongolia Yili Industrial Group Co Ltd
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/19—Dairy proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C21/00—Whey; Whey preparations
    • A23C21/08—Whey; Whey preparations containing other organic additives, e.g. vegetable or animal products
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C21/00—Whey; Whey preparations
    • A23C21/10—Whey; Whey preparations containing inorganic additives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
    • A23C9/1522—Inorganic additives, e.g. minerals, trace elements; Chlorination or fluoridation of milk; Organic salts or complexes of metals other than natrium or kalium; Calcium enrichment of milk
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
    • A23C9/1526—Amino acids; Peptides; Protein hydrolysates; Nucleic acids; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
    • A23C9/156—Flavoured milk preparations ; Addition of fruits, vegetables, sugars, sugar alcohols or sweeteners
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
    • A23C9/158—Milk preparations; Milk powder or milk powder preparations containing additives containing vitamins or antibiotics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/16—Agglomerating or granulating milk powder; Making instant milk powder; Products obtained thereby
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/15—Vitamins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/15—Vitamins
    • A23L33/155—Vitamins A or D
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/16—Inorganic salts, minerals or trace elements
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Abstract

The invention provides a composition and application thereofAnd food. The composition contains protein, calcium, and vitamin D3Vitamin K2Wherein: the mass ratio of protein to calcium is 14:1-23: 1; vitamin D3The mass ratio of the calcium to the calcium is 1: 1.0X 105‑4.0×105(ii) a Vitamin K2The mass ratio of the calcium to the calcium is 1: 1.5X 104‑5.0×104. The invention also provides application of the composition in preparing food with the function of promoting calcium absorption or promoting bone health and food containing the composition. The invention combines and prefers calcium and vitamin D by adjusting the ratio of protein to calcium3+ vitamin K2The composition and the food obtained by the method realize obvious gradual synergistic effect in the aspects of improving physical development, increasing bone calcium content, increasing cortical bone thickness, increasing cortical bone area and the like.

Description

Composition, application thereof and food
Technical Field
The invention relates to a composition, application thereof and food, and belongs to the technical field of dairy products.
Background
Childhood is the peak period of skeletal development, and calcium and protein are important nutrients for maintaining skeletal health. Calcium intake can affect the effect of dietary proteins on bone health in children, and in the case of sufficient calcium intake, increasing dietary proteins can increase bone density (BMD), Bone Mineral Content (BMC), with potentially positive effects on bone health. The proper ratio of calcium to protein in the diet can promote the growth and development of children and teenagers.
At present, the dietary protein intake of children and teenagers in China is basically met, but the situation of serious calcium intake insufficiency exists. Milk is an important high-quality food source of calcium and protein, but the average daily intake of milk and milk products of each person of children aged 6-17 years in China is 34.5g, which only accounts for about 1/10 of the recommended amount. The average calcium intake of children aged 3-6, 7-9 and 10-13 years in China is only 318mg, 334mg and 390mg, and the recommended intake (RNI) values are 800mg, 1000mg and 1200mg respectively. The protein intake of children 3-6 years old, 7-9 years old and 10-13 years old reaches 41g, 51g and 60g, the weight ratio of protein/calcium intake of children in China reaches 129:1, 153:1 and 154:1, which is far more than the ratio of proper protein and calcium, and is not beneficial to bone health and physique development. This is related to the dietary pattern of low milk consumption, high fat and high protein meat food or high carbohydrate staple food intake for children in China. No report of a protein/calcium ratio dietary nutrition scheme beneficial to bone health and physique development in childhood aiming at Chinese children dietary conditions is seen at present.
Based on the dietary pattern of Chinese children, the adjustment of the nutritional ingredients of milk is still urgently needed at present to provide formula milk powder for improving bone health, so that a more balanced nutritional dietary pattern is provided for Chinese children.
Disclosure of Invention
In order to solve the above technical problems, the present invention provides a composition comprising protein, calcium, and vitamin D3Vitamin K2Has the function of promoting calcium absorption or bone health.
In order to achieve the above objects, the present invention provides a composition comprising protein, calcium, vitamin D3Vitamin K2Wherein:
the mass ratio of protein to calcium is 14:1-23: 1;
vitamin D3The mass ratio of the calcium to the calcium is 1: 1.0X 105-4.0×105(preferably, the mass ratio of the two is 1: 1.5X 10)5-2.0×105)；
Vitamin K2The mass ratio of the calcium to the calcium is 1: 1.5X 104-5.0×104(preferably, the mass ratio of the two is 1: 1.5X 10)4-3.5×104)。
In the above composition, preferably, vitamin D3With vitamin K2The mass ratio of (1): 3-10, preferably 1: 5-10.
In the above composition, the mass ratio of protein to calcium is 14:1-23:1, specifically, may be 14: 1. 15: 1. 16: 1. 17: 1. 18: 1. 19: 1. 20: 1. 21: 1. 22: 1. 23:1, or a range consisting of one of these specific ratios as an upper limit and the other as a lower limit, for example 15: 1-22: 1. 16: 1-21: 1. 17: 1-20: 1. 18: 1-19: 1, etc.
In the above composition, vitamin D3The mass ratio of the calcium to the calcium is 1: (1.0X 10)5-4.0×105) Specifically, the ratio of 1: 1.0X 105、1：1.1×105、1：1.2×105、1：1.3×105、1：1.4×105、1：1.5×105、1：1.6×105、1：1.7×105、1：1.8×105、1：1.9×105、1：2.0×105、1：2.1×105、1：2.2×105、1：2.3×105、1：2.4×105、1：2.5×105、1：2.6×105、1：2.7×105、1：2.8×105、1：2.9×105、1：3.0×105、1：3.1×105、1：3.2×105、1：3.3×105、1：3.4×105、1：3.5×105、1：3.6×105、1：3.7×105、1：3.8×105、1：3.9×105、1：4.0×105Or a range in which one of these specific ratios is an upper limit and the other is a lower limit, for example, 1: (1.1X 10)5-3.9×105)、1：(1.2×105-3.8×105)、1：(1.3×105-3.7×105)、1：(1.4×105-3.6×105)、1：(1.5×105-3.5×105)、1：(1.6×105-3.4×105)、1：(1.7×105-3.3×105)、1：(1.8×105-3.2×105)、1：(1.9×105-3.1×105)、1：(2.0×105-3.0×105)、1：(2.1×105-2.9×105)、1：(2.2×105-2.8×105)、1：(2.3×105-2.7×105)、1：(2.4×105-2.6×105)。
In the above composition, vitamin K2The mass ratio of the calcium to the calcium is 1: (1.5X 10)4-5.0×104) Specifically, the ratio of 1: 1.5X 104、1：1.6×104、1：1.7×104、1：1.8×104、1：1.9×104、1：2.0×104、1：2.1×104、1：2.2×104、1：2.3×104、1：2.4×104、1：2.5×104、1：2.6×104、1：2.7×104、1：2.8×104、1：2.9×104、1：3.0×104、1：3.1×104、1：3.2×104、1：3.3×104、1：3.4×104、1：3.5×104、1：3.6×104、1：3.7×104、1：3.8×104、1：3.9×104、1：4.0×104、1：4.1×104、1：4.2×104、1：4.3×104、1：4.4×104、1：4.5×104、1：4.6×104、1：4.7×104、1：4.8×104、1：4.9×104And 1: 5.0 × 10, or a range in which one of these specific ratios is an upper limit and the other is a lower limit, for example, 1: (1.6X 10)4-4.9×104)、1：(1.7×104-4.8×104)、1：(4.7×104-3.2×104)、1：(1.9×104-4.6×104)、1：(2.0×104-4.5×104)、1：(2.1×104-4.4×104)、1：(2.2×104-4.3×104)、1：(2.3×104-4.2×104)、1：(2.4×104-4.1×104)、1：(2.5×104-4.0×104)、1：(2.6×104-3.9×104)、1：(2.7×104-3.8×104)、1：(2.8×104-3.7×104)、1：(2.9×104-3.6×104)、1：(3.0×104-3.5×104)、1：(3.1×104-3.4×104)、1：(3.2×104-3.3×104)。
In the above composition, preferably, the raw material for providing protein includes, but is not limited to, one or a combination of two or more of raw milk, desalted whey powder, and α -lactalbumin powder.
In the above compositions, preferably, the source of calcium includes, but is not limited to, calcium carbonate, and may also include all suitable sources of calcium, such as: calcium acetate, calcium caseinate, calcium chloride, calcium citrate malate, calcium gluconate, calcium lactate, calcium malate, calcium hydrogen phosphate, calcium dihydrogen phosphate, calcium sulfate, calcium ascorbate, calcium glycerophosphate, milk calcium, etc.
The composition provided by the invention has good effects on promoting calcium absorption and promoting bone health, wherein the bone health promotion comprises improvement of physical development, increase of bone calcium content, increase of cortical bone thickness, increase of cortical bone area, reduction of CTX-I, reduction of ratio of uncarboxylated osteocalcin to carboxylated osteocalcin and the like.
Therefore, the invention also provides the application of the composition in preparing food with the function of promoting calcium absorption or bone health.
According to a specific embodiment of the present invention, preferably, the promotion of bone health comprises one or a combination of two or more of improving physical development of the animal, increasing bone calcium content, increasing cortical bone thickness, increasing cortical bone area, decreasing CTX-I, decreasing the ratio of uncarboxylated osteocalcin to carboxylated osteocalcin.
According to a particular embodiment of the invention, preferably, the animal is selected from the group consisting of a growth-stage animal and an animal with insufficient calcium intake during other growth stages than the growth stage; more preferably, the animal is a mammal. Further preferably, the animal is a child in a growing stage.
According to a particular embodiment of the invention, preferably, the food product is a liquid dairy product, a milk powder, a milk tablet or a nutritional supplement; more preferably, the milk powder is a formula, such as an infant formula, a toddler formula or a child formula.
The invention also provides a food containing the composition.
According to a particular embodiment of the invention, preferably, the food product is a liquid dairy product, a milk powder, a milk tablet or a nutritional supplement; more preferably, the milk powder is a formula, such as an infant formula, a toddler formula or a child formula.
According to the specific embodiment of the present invention, preferably, the content of calcium is 600-1800mg/100g (preferably 1000-1500mg/100g) based on the weight of the formula powder; vitamin D3The content of (b) is 3-18.9. mu.g/100 g (preferably 4-9. mu.g/100 g), vitamin K2The content of (b) is 18 to 135. mu.g/100 g (preferably 30 to 80. mu.g/100 g).
According to the specific embodiment of the invention, preferably, the content of the compound vitamin is 0.1-0.45% by weight of the formula milk powder as 100%; the vitamin D3Vitamin K2Is provided by compound vitamin, and the vitamin D is calculated by the weight of the compound vitamin3The content of (A) is 30-42 mug/g, vitamin K2The content of (b) is 180-300. mu.g/g.
According to a particular embodiment of the invention, preferably, the formula contains 17% to 26% protein by mass.
According to a specific embodiment of the present invention, preferably, the formula contains, by mass: 17 to 26 percent of protein, 12 to 20 percent of fat, 2 to 8 percent of dietary fiber, 48 to 55 percent of carbohydrate, 0.1 to 0.45 percent of compound vitamin and 1.5 to 1.7 percent of compound mineral.
According to a particular embodiment of the invention, in the above formula, the fat is mainly milk fat, derived from milk-based raw materials.
According to a specific embodiment of the present invention, preferably, the raw material for providing dietary fiber comprises one or a combination of two or more of galacto-oligosaccharide, resistant dextrin, inulin, and the like.
According to a particular embodiment of the invention, preferably, in the above formula, the ratio of protein to calcium ranges from 14:1 to 23: 1.
According to a particular embodiment of the invention, preferably, in the above formula, each gram of mineral comprises 70-120mg of iron and 60-100mg of zinc.
According to a specific embodiment of the present invention, preferably, in the above formula, each gram of the compound vitamin comprises: vitamin A: 1300-; vitamin D3：30-42μg；Vitamin B1: 2500-; vitamin C: 200-330 mg; vitamin E: 9-15mg of alpha-TE; folic acid: 700-; pantothenic acid: 8000-11000 mu g; nicotinic acid: 9-15 mg; vitamin K2: 180-300 μ g; taurine: 100-200 mg; lutein: 800-1100 mu.g.
According to a particular embodiment of the invention, preferably, the formula also comprises choline, such as choline chloride.
According to a specific embodiment of the present invention, preferably, the content of choline chloride is 900-1500g/1000kg of formula milk powder.
The invention provides a preparation method of the formula milk powder, which comprises the following steps: preparing materials, homogenizing, sterilizing, concentrating, and spray drying.
According to a specific embodiment of the invention, the preparation steps of the formula milk powder are as follows:
1) mixing materials: adding desalted whey powder, galacto-oligosaccharide, alpha-lactalbumin powder, phospholipid and other raw materials and a nutrient bag (comprising a vitamin bag, a choline chloride bag, a calcium powder bag and a mineral bag) into raw milk to mix to obtain a feed liquid, wherein preferably, the mixing concentration is 20-30%, the mixing temperature is 40-50 ℃, and the mixing time is 35-60 minutes;
2) homogenizing: homogenizing the feed liquid, preferably, the homogenizing pressure is 20-140MPa, and the homogenizing temperature is 50-60 ℃;
3) pasteurization: sterilizing the homogenized feed liquid, preferably at 92-95 deg.C for 10-15 s;
4) concentration: concentrating the sterilized feed liquid, preferably at 47-55 deg.C, and the concentration of feed liquid is 45-55%;
5) spray drying: spray drying the concentrated feed liquid to obtain uniformly dried powder, preferably, the air inlet temperature is 165-170 ℃, and the air exhaust temperature is 70-80 ℃.
6) Dry mixing: weighing 1000kg of milk powder obtained by spray drying, weighing dry-mixed raw materials such as DHA, ARA, lactoferrin and the like, and uniformly stirring for 15-25 minutes in a dry mixer at a constant speed of 50-120rpm/min to obtain uniformly dried powder, namely the formula milk powder.
The invention combines and prefers calcium and vitamin D by adjusting the ratio of protein to calcium3+ vitamin K2The combination provides a dietary nutrition improvement scheme with high accessibility, and the obtained composition, food and formula milk powder realize obvious progressive synergistic effects in the aspects of improving the physical development of children or animals in the growth period, increasing the bone calcium content, increasing the cortical bone thickness, increasing the cortical bone area, reducing CTX-I (type I collagen cross-linked carboxyl terminal peptide), reducing the ratio of uncarboxylated osteocalcin/carboxylated osteocalcin and the like.
Drawings
FIG. 1a is a schematic representation of bone density imaging in rats.
FIG. 1b is an image of rat Micro-CT.
FIG. 2 is a cross-sectional Micro-CT representative image of the distal end of each set of femurs.
FIG. 3 shows the effect of the test substance on the body weight of rats during the experiment.
Detailed Description
The technical solutions of the present invention will be described in detail below in order to clearly understand the technical features, objects, and advantages of the present invention, but the present invention should not be construed as limiting the implementable scope of the present invention.
Example 1
The embodiment provides a formula milk powder, which comprises the following components:
3800kg of raw milk, 420kg of desalted whey powder, 100kg of galacto-oligosaccharide, 18kg of anhydrous cream, 12kg of alpha-lactalbumin powder and 2kg of phospholipid;
16kg of nutrients (including a compound vitamin bag, a compound mineral bag, a choline bag and a compound calcium carbonate bag), wherein the calcium content is 1200mg/100g and the vitamin D is calculated according to the mass of the formula milk powder3The content of vitamin K is 8 mug/100 g2The content is 50 mu g/100 g; lutein 150 μ g/100g, zeaxanthin 100 μ g/100g, yeast beta-glucan 50mg/100g, DHA 50mg/100g, ARA 60mg/100g, lactoferrin 30mg/100g, vitamin A, vitamin B1The addition of other nutrients such as vitamin C, folic acid, pantothenic acid and the like is the conventional addition amount within the allowed range of the regulation;
the formula milk powder comprises the following main nutrient components:
energy 1897kJ/100g, protein content 19.0%, fat content 18.0%, dietary fiber content 3%, carbohydrate content 51%, and the ratio of protein to calcium 15.8: 1.
Example 2
The embodiment provides a formula milk powder, which comprises the following components:
3800kg of raw milk, 440kg of desalted whey powder, 100kg of galacto-oligosaccharide, 20kg of alpha-lactalbumin powder and 2kg of phospholipid;
17kg of nutrients (including a compound vitamin bag, a compound mineral bag, a choline bag and a compound calcium carbonate bag), wherein the calcium content is 1250mg/100g and the vitamin D is calculated according to the mass of the formula milk powder3The content of vitamin K is 7 mug/100 g2The content is 38 mu g/100 g; lutein 150 μ g/100g, zeaxanthin 100 μ g/100g, yeast beta-glucan 50mg/100g, DHA 50mg/100g, ARA 60mg/100g, lactoferrin 30mg/100g, vitamin A, vitamin B1The addition of other nutrients such as vitamin C, folic acid, pantothenic acid and the like is the conventional addition amount within the allowed range of the regulation;
the formula milk powder comprises the following main nutritional components:
energy 1987kJ/100g, protein content 19%, fat content 18%, dietary fiber content 3%, carbohydrate content 51%, protein to calcium ratio 15.2: 1.
Example 3
The embodiment provides a formula milk powder, which comprises the following components:
2000kg of raw milk, 530kg of desalted whey powder, 100kg of galacto-oligosaccharide, 110kg of anhydrous cream, 25kg of alpha-lactalbumin powder and 2kg of phospholipid;
nutrients (12kg, including a compound vitamin bag, a compound mineral bag, a choline bag and a compound calcium carbonate bag), wherein the calcium content is 610mg/100g and the vitamin D is calculated according to the mass of the formula milk powder3The content of vitamin K is 4 mug/100 g2The content is 40 mu g/100 g; lutein 150 μ g/100g, zeaxanthin 100 μ g/100g, yeast beta-glucan 50mg/100g, DHA 50mg/100gg. ARA 60mg/100g, lactoferrin 30mg/100g, vitamin A, vitamin B1The addition of other nutrients such as vitamin C, folic acid, pantothenic acid and the like is the conventional addition amount within the allowed range of the regulation;
the formula milk powder comprises the following main nutrient components:
energy 1908kJ/100g, protein content 14%, fat content 19%, dietary fiber content 4%, carbohydrate content 55%, protein to calcium ratio 23: 1.
The formula of examples 1-3 was prepared by the following steps:
1) mixing materials: adding powdery raw materials into raw milk to mix, so as to obtain a feed liquid, wherein the mixing concentration is 25%, the mixing temperature is 47 ℃, and the mixing time is 40 minutes;
2) homogenizing: homogenizing the feed liquid at 20-140MPa and 55 deg.C;
3) pasteurization: sterilizing the homogenized feed liquid at 95 deg.C for 13 s;
4) concentration: concentrating the sterilized feed liquid at 50 deg.C to give 48% feed liquid;
5) spray drying: spray drying the concentrated feed liquid to obtain uniformly dried powder, wherein the air inlet temperature is 165-170 ℃, and the air exhaust temperature is 75 ℃;
6) dry mixing: weighing 1000kg of milk powder obtained by spray drying, weighing dry-mixed raw materials such as DHA, ARA, lactoferrin and the like, and uniformly stirring for 20 minutes in a dry mixer at a constant speed of 50-120rpm/min to obtain uniformly dried powder, namely the formula milk powder.
Test animals and groups:
SPF-grade weaning SD rats 144 male, weighing 45-65 g, purchased from Beijing Wittingle laboratory animal technology, Inc., animal qualification number SCXK (Jing) 2016-. The animals are raised in the barrier level animal house of the national institutes of public health of western Sichuan university, the qualification number SYXK (Sichuan) 2018-. During feeding, the animals are ensured to drink water freely and eat fully, and the drinking water is sterilized pure water. The rats are raised in a cage with a stainless steel wire at the bottom and an animal roomThe interior of the box body is kept in a quiet, clean, ventilated and proper illumination state, the temperature is 20-26 ℃, the humidity is 40% -70%, and the light and shade alternating period is 12 h. Specific experimental groups and calcium, vitamin D3Vitamin K2The contents of (A) are shown in Table 1-1.
TABLE 1-1 Experimental groups
Figure BDA0003520791160000071
Figure BDA0003520791160000081
The rats were fed with AIN-93G standard feed (Nantong Denofil feed science and technology Co., Ltd., production license number: Su-sho ((2019) 06092)) for 2 weeks, and the rats were randomly divided into 5 groups of 12 animals each with calcium and vitamin D3The administration is carried out by mixing and feeding, and the formula of each group of feed is shown in the table 1-2. Intervention of MK adopts a gastric lavage mode to add vitamin K2The powder is uniformly dissolved in corn oil. Vitamin K at 9 o' clock in the morning every day2Performing intragastric lavage according to corresponding dose in the corresponding experimental group for intervention without intake of vitamin K2The control group and the experimental groups of the (1) are filled with corn oil with the same dosage. The intragastric volume is 0.5mL/100 g. All dressing and gavage treatments were continued for 93 days after the animals had been acclimatized. During the period, the weight is weighed once every 7 days, and the gavage amount is adjusted according to the change of the weight. Animals were sacrificed on day 93 of the experiment and tested for each index.
Table 1-2 feed formulations for each experimental group
Item Feed 1 Feed 2 Feed 3
Corn starch, g/kg 422.486 422.486 422.486
Casein, g/kg 200.000 200.000 200.000
Dextrin, g/kg 132.000 132.000 132.000
Sucrose, g/kg 100.000 100.000 100.000
Soybean oil, g/kg 70.000 70.000 70.000
L-cystine, g/kg 3.000 3.000 3.000
Cellulose, g/kg 25.000 25.000 25.000
Calcium-free mineral mix package, g/kg 22.505 22.505 22.505
Calcium carbonate, g/kg 3.750 12.500 9.375
Is free of D3Vitamin mix pack, g/kg 10.000 10.000 10.000
Vitamin D3，g/kg 0.010 0.010 0.0148
Tartaric acid, g/kg 2.500 2.500 2.500
Tert-butyl hydroquinone, g/kg 0.014 0.014 0.014
In total, g 991 1000 997
Experimental examples 1, 2, 3 of animal experiments used feed 1, 2, 3, respectively; experimental examples 4, 5 and 6 Using feed 3, different doses of VK were added2The specific additive amounts are shown in Table 1-1.
Vitamin K in animal experiments2The dose is the total dose of all sources of the whole experimental animal diet, and the children over 3 years old do not only eat milk powder, but also vitamin K in the diet of the children2The source of the milk comprises vegetables and fermented food, and the milk powder for children is used for supplementing vitamin K2Good food carrier, therefore, vitamin K in milk powder2The dosage design of (A) can be 1/6-1/3 of animal experiments.
Experimental methods
Animal handling and sample collection:
after the last gavage, the rats were fasted for 16h and water was freely available. After anesthetization, the abdominal veins were bled. The rats were sacrificed by cervical dislocation. The blood sample is centrifuged at 3500r/min for 10min at 4 ℃ to separate serum, and then supernatant is extracted. Serum was stored at-80 ℃ and removed when tested.
After the rats were sacrificed, bilateral femurs were separated, muscles and soft tissues attached to the bone surfaces were removed, and bilateral femurs were exfoliated. The left femur was oven dried in a 105 ℃ oven. The right femur was fixed with 4% paraformaldehyde for 36h, then soaked in 70% alcohol and stored in a refrigerator at 4 ℃ until CT scan.
The effect of the test substances on the body weight of rats (Mean + SD) (g) during the experiment is shown in fig. 3.
Femur weight, femur length, bone calcium and bone density determination:
femoral weight and femoral length determination: after the left femur was dried in an oven at 105 ℃ to constant weight, the dry weight of the femur was recorded and the length of the femur was measured.
Bone calcium assay: the left femur dried to constant weight is placed in a crucible,heating with small fire, charring to no smoke, transferring into muffle furnace, and ashing at 550 deg.C for 3-4 h. Cooling, taking out, digesting with concentrated nitric acid, fixing the volume to 25mL, taking 40 mu L to 10mL of colorimetric tube, adding 1mL of 20% lanthanum nitrate solution, fixing the volume to 10mL, shaking up, and measuring by using a flame atomic absorption spectrometer.
Bone Density determination: the bone density of the middle end of each rat femur was measured by a German Bruker DXS PRO type small animal full-automatic high resolution X-ray machine for the rat right femur after Micro-CT scanning, 10 rat femurs were imaged at the same time each time, the pixel resolution was 10 μm, the X-ray lamp voltage was 45kVp, and the exposure time was set to 1 min. The test results are shown in FIG. 1 a.
Micro-CTDetection of
The right femur of the rat was scanned by a small animal living Micro-CT Imaging System from PerkinElmer, USA, and analyzed by Quantum GX Micro CT Imaging System software. The scan parameters are set as follows: the field of view was set at 36mm, voxel 72 μm, energy 80kV, intensity 100 μ A, scan time 14 min. After three-dimensional reconstruction of scanning data, 512 pictures with different cross sections are obtained by scanning the same sample, and a region of interest (ROI) is set to be 50 pictures from 0.5mm below a growth layer). And (4) reconstructing the scanning data by using software and then analyzing parameters. The bone density was corrected using a resin-embedded hydroxyapatite solid calibration model from PerkinElmer, and the cortical, cancellous, and all bone densities in the ROI were calculated separately. Other analysis parameters include bone volume fraction (BV/TV), trabecular thickness (tb.th), trabecular number (tb.n), and trabecular separation (tb.sp). FIG. 1b is an image of rat Micro-CT, wherein FIG. 1b is a three-view screenshot of SD rat 5-8 during scanning, D is a cross-sectional view, L is a sagittal view, and T is a cross-sectional view. Representative images of cross-sectional Micro-CT of the distal ends of each set of femurs are shown in fig. 2.
Serum index detection
Serum carboxylated Osteocalcin (Gla-ostoclcin, Gla-OC) and uncarboxylated Osteocalcin (Glu-ostoclcin, Glu-OC) were tested using a sandwich enzyme-linked immunosorbent assay and the percentage of uncarboxylated Osteocalcin to total Osteocalcin was calculated. Type I collagen cross-linked carboxy-terminal peptide (Type I collagen cross-linked C-terminal peptide, CTX-I) was tested using an immunoadsorption assay kit.
Data processing and result determination
All data are expressed as Mean ± standard deviation (Mean ± SD). One-way anova was performed using SPSS 26.0 statistical software. Before analysis, the homogeneity of variance is tested, and the F value is calculated. If the F value is less than F0.05, P is more than 0.05, the mean difference of each group is not statistically significant; if the F value is greater than F0.05 and P is less than 0.05, the difference between the mean values of the groups is statistically significant. Mean pairwise comparisons between multiple experimental and control groups were analyzed using LSD except for Dunnett-t for body weight.
The Micro-CT scan results for each set of femoral microstructures are shown in table 2. The effect of the femoral normative index for each test subject is shown in table 3. The Micro-CT scan results of femoral cortical bone of each group of rats are shown in table 4. The results of vitamin K and bone metabolism indices are shown in table 5.
TABLE 2 Micro-CT scan results for femoral bone microstructures (Mean + SD)
Figure BDA0003520791160000101
TABLE 3 Effect of the femoral Standard indices of Each test subject (Mean + SD)
Group of Femur weight/(g) Length of femur/(mm) Bone density/(g/cm)3) Bone calcium content (mg/g)
Experimental example 1 0.725±0.068d 35.92±0.77 0.945±0.056d 144.33±12.53d
Experimental example 2 0.966±0.090ab 36.54±0.89 1.054±0.073a 214.77±17.23abc
Experimental example 3 0.897±0.095c 36.19±1.02 1.000±0.043bc 201.47±24.31c
Experimental example 4 0.924±0.058bc 36.14±2.29 1.008±0.051ab 206.86±12.52bc
Experimental example 5 0.938±0.061abc 36.83±0.85 1.014±0.053ab 202.98±13.07c
Experimental example 6 0.975±0.113ab 36.63±0.56 1.019±0.075ab 223.88±20.52a
TABLE 4 Micro-CT scan results (Mean + SD) of femoral cortical bone of rats in each group
Numbering Grouping Ct.Ar(mm2) Ct.Th/(μm3)
Experimental example 1 Low calcium group 0.40±0.05d 6.03±0.75e
Experimental example 2 Normal group 0.56±0.04a 9.06±0.85a
Experimental example 3 Strengthening group 0.51±0.05c 7.77±0.99d
Experimental example 4 Low MK group 0.52±0.03bc 8.00±0.41cd
Experimental example 5 Group of middle MK 0.53±0.03abc 8.26±0.4bcd
Experimental example 6 High MK group 0.56±0.06a 8.74±0.94ab
Note: the different letters indicate that the groups are statistically significant compared to each other, P < 0.05. Tt. ar, total cortical bone area (total cortical bone area); ct.ar, cortical bone area (cortical bone area); th, cortical bone thickness (cortical thickness); BMD, bone mineral density (bone mineral density).
TABLE 5 results of vitamin K and bone metabolism indices (Mean + SD)
Figure BDA0003520791160000111
Note: the different letters indicate that the groups are statistically significant compared to each other, P < 0.05. CTX-I, Type I collagen cross-linked carboxyl-terminal peptide (Type I collagen cross-linked C-terminal peptide).
From the above experimental results it can be seen that:
the high calcium group provided by the invention has an effect of improving the bone health of animals in a growth period under the condition of low calcium nutrition, and is shown in an experimental example 2. The calcium dose in the high calcium group was supplemented to the recommended ideal dose for the animals in the growth stage by experimental example 2, and it can be seen from the above experimental results that: the effect of improving bone health of animals in the growth period can be achieved by adjusting the ratio of protein to calcium.
The calcium and the vitamin D provided by the invention3The composition has an effect of improving the bone health of animals growing in a low-calcium nutritional state, see experimental example 3, wherein the dosage of calcium supplemented by the scheme of the experimental example 3 is 3/4 of the experimental example 2, and vitamin D3The dosage of (A) is 1.5 times that of experimental examples 1 and 2, and according to the experimental results, the following can be seen: fortification of vitamin D within a range based on the protein/calcium ratio range of the present invention3Can improve the bone health of animals in the growth period.
The calcium and the vitamin D provided by the invention3+ Low dose vitamin K2For the improvement of bone health in animals with low calcium nutrition in growth stage, see experimental example 4. The dosage of calcium after supplementation was 3/4, vitamin D of Experimental example 2, by the protocol of Experimental example 43The dosage of (A) is 1.5 times that of experimental examples 1 and 2, and it can be seen from the above experimental results that: strengthening vitamin D in a certain range on the basis of the protein/calcium ratio range protected by the invention3And low dose of vitamin K2The effects of improving the bone health and the physique development of the animals in the growth period can be further realized on the basis of the experimental example 3.
The calcium and the vitamin D provided by the invention3+ Low dose vitamin K2For the improvement of bone health in animals with low calcium nutrition in growth stage, see experimental example 5. The dosage of calcium after supplementation was 3/4 of Experimental example 2, vitamin D, by the nutritional regimen of Experimental example 53The dosage of (A) is 1.5 times that of experimental examples 1 and 2, and it can be seen from the above experimental results that: protein to calcium ratio in the protection of the inventionOn the basis of the range of the examples, the vitamin D is enhanced within a certain range3Vitamin K with moderate dosage2The effects of improving the bone health and the physique development of the animals in the growth period can be further realized on the basis of the experimental example 3.
The calcium and the vitamin D provided by the invention3+ high dose vitamin K2For the improvement of bone health in animals with low calcium nutrition in growth stage, see experimental example 6. The dosage of calcium after supplementation was 3/4, vitamin D, from Experimental example 6, according to the nutritional regimen3The dosage of (A) is 1.5 times that of experimental examples 1 and 2, and it can be seen from the above experimental results that: fortification of vitamin D within a range based on the protein/calcium ratio range protected by the present invention3And high dose of vitamin K2Can further realize the functions of improving the bone health and the physique development of the animals in the growth period on the basis of the experimental example 3, and further expands the vitamin K2The protective scope of (1).
Through the comparison of the groups, the invention realizes the technical effects of obviously increasing the level by level in the aspects of improving the physical development of animals in the growth period, increasing the bone calcium content, increasing the cortical bone thickness, increasing the cortical bone area, reducing CTX-I (type I collagen cross-linked carboxyl terminal peptide) and reducing the ratio of uncarboxylated osteocalcin/carboxylated osteocalcin.
According to the above experimental results, it can be seen that: in addition to improving bone health in the long-term population, i.e., children, the present invention may also be used to improve bone health in other populations with calcium malnutrition. For example: the average calcium intake of middle-aged and aged people 50-64 years and 65 years in China is 415mg and 409 mg/day, the RNI value of calcium of people in the age group is 1000mg, which means that the calcium intake condition of the people only reaches about 1/3 of the required amount, and the calcium intake condition is matched with the nutritional condition of the low-calcium group established by the invention.

Claims (11)

1. A composition comprises protein, calcium, and vitamin D3Vitamin K2Wherein:
the mass ratio of the protein to the calcium is 14:1-23: 1;
the vitamin D3The mass ratio of the calcium to the calcium is 1: 1.0X 105-4.0×105Preferably, the mass ratio of the two is 1: 1.5X 105-2.0×105；
The vitamin K2The mass ratio of the calcium to the calcium is 1: 1.5X 104-5.0×104Preferably, the mass ratio of the two is 1: 1.5X 104-3.5×104。
2. The composition of claim 1, wherein said vitamin D3With said vitamin K2The mass ratio of (1): 3-10, preferably, the mass ratio of the two is 1: 5-10.
3. The composition of claim 1, wherein the protein-providing material comprises one or a combination of two or more of raw milk, desalted whey powder, and alpha-lactalbumin powder.
4. Use of the composition of any one of claims 1 to 3 for the preparation of a food product having the function of promoting calcium absorption or promoting bone health.
5. The use of claim 4, wherein the promotion of bone health comprises one or a combination of two or more of improving physical development, increasing bone calcium content, increasing cortical bone thickness, increasing cortical bone area, decreasing CTX-I, decreasing the ratio of uncarboxylated osteocalcin to carboxylated osteocalcin in the animal;
preferably, the animal is a child in a growth stage.
6. Use according to claim 4, wherein the food product is a liquid dairy product, a milk powder, a milk tablet or a nutritional supplement; preferably, the milk powder is a formula, more preferably an infant formula, a toddler formula or a child formula.
7. A food product comprising the composition of any one of claims 1-3; preferably, the food product is a liquid dairy product, a milk powder, a milk tablet or a nutritional supplement.
8. The food product of claim 7, wherein the milk powder is a formula;
preferably, the content of calcium is 600-1800mg/100g, more preferably, the content of calcium is 1000-1500mg/100 g; the vitamin D3The content of (b) is 3-18.9 mug/100 g, preferably 4-9 mug/100 g; vitamin K2The content of (b) is 18 to 135. mu.g/100 g, preferably 30 to 80. mu.g/100 g.
9. The food of claim 8, wherein the content of the compound vitamin is 0.1-0.45% by weight of the formula milk powder as 100%;
the vitamin D3Vitamin K2Is provided by compound vitamin, and the vitamin D is calculated by the weight of the compound vitamin3The content of (b) is 30-42 mug/g, vitamin K2The content of (b) is 180-300. mu.g/g.
10. Food product according to claim 8 or 9, wherein the formula contains 17-26% protein by mass.
11. Food product according to any one of claims 8-10, wherein the formula contains, in mass percent:
17-26% of protein, 12-20% of fat, 2-8% of dietary fiber, 48-55% of carbohydrate, 0.1-0.45% of compound vitamin and 1.5-1.7% of compound mineral;
preferably, the raw material for providing the dietary fiber comprises one or more of galacto-oligosaccharide, resistant dextrin and inulin;
preferably, each gram of mineral comprises 70-120mg of iron and 60-100mg of zinc;
preferably, each gram of compound vitamin comprises:
vitamin A: 1300 gradient 2000. mu.gRE
Vitamin D3：30-42μg
Vitamin B1：2500-4200μg
Vitamin C: 200 and 330mg
Vitamin E: 9-15mg of alpha-TE
Folic acid: 700-1000 μ g
Pantothenic acid: 8000-11000 mu g
Nicotinic acid: 9-15mg
Vitamin K2：180-300μg
Taurine: 100-200mg
Lutein: 800-1100 mu.g;
preferably, the formula further comprises choline, more preferably, the choline is choline chloride; further preferably, the content of the choline chloride is 900-1500g/1000kg of formula milk powder.
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