Anthocyanin ingredient containing milk fat globules and preparation method

Abstract

The invention discloses an anthocyanin ingredient containing milk fat globules and a preparation method thereof.The preparation method comprises the specific steps that the milk fat globules are extracted from fresh raw milk, anthocyanin and the milk fat globules are added into deionized water to be emulsified, and the anthocyanin emulsion type ingredient containing the milk fat globules is obtained; wherein anthocyanin in the ingredients and a fat membrane of the milk fat globules form a specific structure, and the specific structure is rich in long-chain unsaturated fatty acid and medium-chain fatty acid in a specific ratio of 1: (7-10) after being digested by in-vitro simulated gastrointestinal fluid. The anthocyanin ingredient containing the milk fat globules is a novel stable emulsion taking the natural milk fat globules as an anthocyanin carrier, is particularly suitable for being used as an ingredient of nutritional and healthy food, and is wide in application range. And the preparation method adopts a probe type ultrasonic combined high-speed shearing technology, so that the obtained emulsion is better in dispersity and stability, simple, efficient, green and safe.

Description

Technical Field

[0001] This invention relates to an anthocyanin ingredient containing milk fat globules and its preparation method, belonging to the field of food science and technology, especially the field of anthocyanin nutritional ingredients. Background Technology

[0002] Polyphenols, as plant-derived secondary metabolites, are widely found in fruits, vegetables, tea, and other foods. Multiple studies have shown that polyphenols can interact with lipids, affecting their digestion, absorption, and metabolism, and may have positive health effects. Anthocyanins are widely found in fruits and vegetables and are an important component of the daily diet, possessing multiple benefits such as cancer prevention, cholesterol reduction, improved sleep, improved vision, enhanced immune function, and anti-aging. With increasing consumer demand for functional foods, anthocyanins or anthocyanin-containing fruits and vegetables are being added as ingredients to dairy products, such as blueberry yogurt and black goji berry yogurt. The addition of anthocyanins not only improves the antioxidant capacity of dairy products but also imparts excellent sensory qualities. Research on anthocyanins and their ingredients has been receiving considerable attention.

[0003] Milk, as a complete nutritional food, contains abundant nutrients such as protein, fat, vitamins, and minerals. Milk fat is one of the main components of milk, typically comprising 3%–5%. Milk fat is primarily composed of triglycerides, with minor components being polar lipids (such as glycerophospholipids and sphingolipids) and cholesterol. Milk fat globules (MFGs) and their encapsulating membranes (MFGMs) are crucial natural structural and functional units in milk. Regarding health functions, numerous studies have revealed the significant potential of MFGMs in promoting infant neurological and cognitive development, improving gut health (e.g., promoting the growth of beneficial bacteria), regulating immunity, and influencing lipid metabolism. The underlying mechanisms are closely related to MFGMs' ability to slow down fat digestion and influence host metabolism through pathways such as gut microbiota. Research indicates that the unique three-layered membrane structure, complex phospholipid composition (such as sphingolipids and phosphatidylcholine), and specific membrane proteins of MFGMs are the material basis for their diverse biological activities. They also demonstrate advantages as natural emulsifiers or carriers of active ingredients in food. Existing patents have reported methods for extracting and preparing milk fat globule membranes (CN202310585228.0), applications of milk fat globule membranes in infant formula (CN202410798506.5), and applications of milk fat globule membrane proteoglycan complexes (CN202211070187.3), but there are no reports of using milk fat globules (membranes) as anthocyanin lipid carriers to construct nutritional ingredients.

[0004] The reported lipid carriers of anthocyanins include liposomes, Pickering emulsions, W1 / O / W2 multiple emulsions, etc., which are generally prepared by methods such as film dispersion, dynamic high-pressure microfluidization, high-pressure homogenization, spontaneous emulsification, and the commonly used oil phase materials include glycerol triester, oleic acid, lecithin, etc.; the water phase materials include soybean protein isolate, whey protein, casein, etc.; the surfactant composition includes Tween, Span, etc.; and the co-surfactant is generally glycerol or ethanol. These systems improve the stability of the system by adding surfactants and co-surfactants, but at the same time, too many exogenous additives are introduced, which affects the subsequent industrial application in food systems. Therefore, the present application uses natural milk fat globules as materials to prepare stable anthocyanin preparations through ultrasonic homogenization and synergistic methods, without introducing too much oil phase, organic reagent, surfactant, etc., which is green and healthy, low in cost, and has a wide industrial application prospect. SUMMARY

[0005] The present application aims to provide an anthocyanin preparation containing milk fat globules and a simple and efficient preparation method thereof, which provides technical support for the development of anthocyanin carriers, the research and development of preparations, and the extended application of milk fat globules.

[0006] The anthocyanin preparation containing milk fat globules and the preparation method thereof are as follows: The anthocyanin preparation containing milk fat globules of the present application comprises milk fat globules, anthocyanins and water, and contains 4-10 g of milk fat globules and 0.15-0.5 g of anthocyanins per 100 g of water.

[0007] The milk fat globules are milk fat globule extracts separated from fresh cow milk, and the anthocyanins are obtained by extracting Lycium ruthenicum Murr from the Xinjiang Tuokexun region, and the main components are petunidin-3-O-rutinoside (coumaroyl) -5-O-glucoside with a content of ≥95% and a small amount of cyanidin-3-O-coumaroyl-glucoside.

[0008] Further, the anthocyanin preparation containing milk fat globules is a stable emulsion with a specific structure formed by the multiple interactions of electrostatic interaction, hydrogen bonding, van der Waals force and hydrophobic interaction between the lipid membrane of Lycium ruthenicum Murr anthocyanins and the milk fat globules of cow milk.

[0009] Further, the anthocyanin preparation containing milk fat globules is rich in long-chain unsaturated fatty acids and medium-chain fatty acids with a specific ratio of 1:7-10 after in vitro simulation of gastrointestinal fluid digestion.

[0010] The preparation method of the anthocyanin preparation containing milk fat globules is characterized by comprising the following preparation steps: (1) Extracting milk fat globules from fresh raw cow milk; (2) The anthocyanin and milk fat globule are added to deionized water for emulsification to obtain an emulsion, which is the anthocyanin preparation containing the milk fat globule.

[0011] In step (1), the milk fat globule extract is obtained by taking fresh raw milk, removing impurities, adjusting the pH to 4.5, collecting the upper cream phase at 4 °C by centrifugation at 3000-8000 rpm, mixing with sterile water, collecting the fat layer by centrifugation at 3000-8000 rpm, then washing the oil phase with deionized water to remove proteins, and repeating the washing 2-3 times to obtain a paste extract, i.e. the milk fat globule.

[0012] In step (2), the black nightshade anthocyanin in claim 2 and the milk fat globule in step (1) are added to deionized water for emulsification, wherein the milk fat globule is added at a mass fraction of 4-10%, the anthocyanin is added at a mass fraction of 0.15-0.5%, and the deionized water is adjusted to a pH of 4.0-5.0.

[0013] Further, preferably, the black nightshade anthocyanin is added at an amount of 0.15-0.45%.

[0014] Further, preferably, the milk fat globule is added at an amount of 5-8%.

[0015] In step (2), the emulsification is first treated with a probe-type ultrasonic instrument (30%, 5s on / 3s off) for 2-5 min, then treated with a high-speed shearing machine at 9000-12000 rpm for 3-10 min, and left to stand for 24 h, and the emulsion has no layering phenomenon, i.e. the emulsion has a particle size of less than 980 nm.

[0016] Further, preferably, the emulsion has a particle size of 140-550 nm.

[0017] Beneficial effects: (1) The preparation method of the anthocyanin preparation containing the milk fat globule of the present application involves centrifugation, water washing and emulsification, which is simple, green and safe, and the anthocyanin preparation containing the milk fat globule is novel.

[0018] (2) The novel nano-scale stable emulsion with the natural milk fat globule as the anthocyanin carrier has a wide application scenario in food.

[0019] (3) The anthocyanin preparation containing the milk fat globule of the present application uses black nightshade anthocyanin which has the effect of synergistic enzyme promoting the release of fatty acids, so that the preparation is rich in specific proportions of long-chain unsaturated fatty acids and medium-chain fatty acids (the ratio of medium-long chain fatty acids is 1:7-10) after in vitro simulation of gastrointestinal fluid digestion, and is particularly suitable as a nutrient and healthy food preparation.

[0020] (4) The anthocyanin preparation containing milk fat globules has the characteristics that anthocyanins are adsorbed on the interface of milk fat globule membranes and acylated groups are inserted into the hydrophobic part of the fat membrane to form a "mosaic" stable structure, and the comprehensive interaction of electrostatic interaction, hydrogen bond interaction, van der Waals force and hydrophobic interaction can significantly improve the stability of the anthocyanin preparation, and compared with traditional anthocyanin emulsion preparation, the anthocyanin preparation has significantly improved stability and functional characteristics of anthocyanins.

[0021] (5) The method for preparing the anthocyanin preparation containing milk fat globules adopts a probe-type ultrasonic combined with high-speed shearing technology, which is shorter in time and higher in efficiency than traditional oscillation method and vortex method, and the obtained emulsion has better dispersibility and stability. DETAILED DESCRIPTION

[0022] The application will be described in detail below in combination with specific examples. However, the application is not limited to the following examples. The methods are all conventional methods unless otherwise specified. The raw materials can be obtained from public commercial channels unless otherwise specified.

[0023] The extraction, separation, purification and identification method of anthocyanins: Lycium ruthenicum Murr. fruit dry from Toksun region of Xinjiang was ground, and hydrochloric acid ethanol (pH 3.0) was added at a material-liquid ratio of 1:30, and ultrasonic extraction was performed for 40 min. The extract was collected by filtration, and the extraction was repeated 3 times. The extract was combined, and the solvent was removed by a rotary evaporator. AB-8 macroporous resin was used for purification, and adsorption equilibrium was achieved after 5 h. Desorption equilibrium time was 4 h, and 75% ethanol-citric acid solution (pH 3.0) was used for desorption. The loading rate was 2.0 mL / min, and the loading volume was 500 mL of crude extract. Elution was performed at a flow rate of 1.0 mL / min using 3 bed volumes of eluent. The eluate was concentrated by rotary evaporation to obtain anthocyanins, and HPLC-ESI-MS was used to identify the extracted anthocyanins. The chromatographic conditions were as follows: Kromasil C18 column (4.6 mm x 250 mm, 5 μm), mobile phase 1% formic acid aqueous solution (A); acetonitrile solution (B), flow rate: 0.4 mL / min; injection volume: 10 μL; column temperature 30 °C, detection wavelength 530 nm. Gradient elution program: 0-35 min, 10%-25% B; 35-45 min, 25%-40% B; 45-46 min, 40%-10% B; 46-50 min, 10%-10% B. Mass spectrometry conditions: ESI ion source, product mode, scanning in positive ion mode, scanning range m / z=50-1100. Ion source nitrogen temperature 35 °C, flow rate 8 L / min; ion source spray voltage 45 psi; sheath gas temperature 400 °C, sheath gas flow rate 12 L / min. Capillary voltage +4000 V; nozzle voltage 500 V. Identification showed that the main components of Lycium ruthenicum anthocyanins were petunidin-3-O-rutinoside (coumaroyl) -5-O-glucoside with a content of ≥95%, and a small amount of cyanidin-3-O-coumaroyl-glucoside.

[0024] Isothermal titration calorimetry detection: The interaction between anthocyanins and milk fat globules was measured using a Microcal VP-ITC isothermal titration calorimeter (Malvern Instruments Ltd., UK). The experimental conditions were set as follows: 2.0 mM milk fat globule membrane solution (0.1 mM PBS, pH 4.0) was loaded into the calorimetric cell, and 40.0 mM anthocyanin solution (0.1 mM PBS, pH 4.0) was filled into the syringe. The program running parameters were as follows: total injection volume 19.0 μL, reaction cell temperature constant 25 °C, stirring speed 750 rpm. The reference power was set to 6 μcal / sec, and the baseline was balanced for 60 seconds. The control experiment was to titrate anthocyanins into PBS buffer (0.1 mM, pH 7.0). The data were processed using the Origin software built-in the instrument, and the corrected values were obtained by subtracting the control group data from the raw experimental data. By fitting the corrected heat flow-time curve (μJ / s vs min) and normalizing, the molar enthalpy change curve (ΔH, kJ / mol) was obtained. The thermodynamic parameters were calculated according to the single-site binding model, including the binding constant (Ka), enthalpy change (ΔH), entropy change (ΔS), and the number of anthocyanin molecules bound per mole of liposome (N). When ΔH < 0, it indicates that the reaction is driven by enthalpy, and usually the interaction is generated by hydrogen bond, ionic bond, and van der Waals force. When TΔS < 0, it indicates that it is an entropy-driven process, and usually hydrophobic interaction occurs.

[0025] In vitro simulated digestion steps: The simulated gastric fluid (SGF) contains 5 g / L pepsin (400-800 U / mg protein), 33.5 g / L NaCl, pH 3.0, and the simulated intestinal fluid (SIF) contains 4 g / L trypsin (250 U / mg protein), 33.25 g / L CaCl2, 26.7 g / L pancreatic lipase (30-90 U / mg protein), 10 g / L porcine bile salt, pH 7.0. 10 mL of milk fat globule-anthocyanin preparation was mixed with 20 mL of SGF, and incubated in a preheated 37 °C shaker at a stirring rate of 200 r / min. After 2 h of incubation, 0.1 mol / L NaOH was added to adjust the pH to 7.0 to inactivate pepsin and stop the gastric digestion stage. Then, 10 mL of the gastric digestion solution was added to 40 mL of SIF in a shake flask, and incubated in a 37 °C shaker at a stirring rate of 200 r / min. The digestion solution was collected at 120 min.

[0026] In vitro digestion product fatty acid composition detection steps: Product extraction and derivatization: First, the pH of the small intestine digestion product was adjusted to below 1.5, mixed solvent (chloroform:methanol = 2:1, v / v) and digestion product were mixed at a volume ratio of 2:1, ultrasonic extraction for 10 min, then centrifugal collection of supernatant organic phase, rotary evaporation to remove organic solvent, obtain lipid phase. Take 1 mL oil sample in 10 mL screw cap glass test tube, continue to add 2 mL of 0.5 mol / L concentration of KOH-CH3OH solution, placed in 65 °C water bath for 30 min for saponification reaction; every 10 min during the period, take out vortex for 10 s. After cooling to room temperature, 2 mL of BF3·C2H5OC2H5 methanol solution (v / v, 1:3) was added, and it was placed in a 70 °C water bath for 10 min for methyl esterification reaction; every 3 min during the period, take out and shake for 30 s. After cooling to room temperature, 2 mL of chromatographic grade n-hexane reagent was added, and after shaking for 3 min, it was allowed to stand for layer separation; the upper liquid was absorbed into a 1.5 mL centrifuge tube, a small amount of anhydrous Na2SO4 solid powder was added, and after shaking for 1 min, it was centrifuged at 5000 r / min for 5 min. The supernatant was taken out, filtered through a 0.22 μm organic filter membrane into a gas chromatography sample vial, sealed with a sealing film, and stored at -18 °C for testing.

[0027] Fatty acid detection: The Thermo Fisher chromatographic column (model: TR-FAME, 60 m x 0.25 mm ID x 0.25 μm) was selected, the column oven initial temperature was 130 °C, maintained for 3 min, then the temperature was raised to 200 °C at a rate of 5 °C / min, maintained for 10 min, finally the temperature was raised to 220 °C at a rate of 2 °C / min, maintained for 3 min, and the program ended. The injection port temperature was 250 °C; the detector temperature was 280 °C. The carrier gas was high-purity nitrogen (≥99.999%), and the flow rate was 1.8 mL / min. The injection volume was 1 μL, and the split ratio was 20.

[0028] Example 1 Fresh raw milk was removed of impurities, the pH was adjusted to 4.5, and the upper cream phase was collected by centrifugation at 3000 rpm at 4 °C. The fat layer was collected by centrifugation after mixing with sterile water, and the oil phase was collected by washing with deionized water to remove proteins. The washing was repeated 3 times to obtain milk fat globules. 100 g of deionized water was prepared, the pH was adjusted to 4.0, 10 g of milk fat globules and 0.5 g of black wolfberry anthocyanins were mixed, and the mixture was first treated by a probe-type ultrasonic instrument (30%, 5 s on / 3 s off) for 2 min, and then treated by a high-speed shear machine at 12000 rpm for 3 min for emulsification. The emulsion particle size was measured by a laser nanoparticle size analyzer to be 905 nm.

[0029] The comprehensive analysis by fluorescence spectroscopy, polarized light spectroscopy, Fourier infrared spectroscopy and isothermal titration calorimetry shows that the black medlar anthocyanins and the lipid membrane of the milk fat globule have multiple interactions such as electrostatic interaction, hydrogen bond interaction, van der Waals force and hydrophobic interaction, and the black medlar anthocyanins are adsorbed on the interface of the milk fat globule membrane, the acylated group is inserted into the hydrophobic part of the lipid membrane to increase the hydrophobicity and membrane fluidity of the lipid membrane, and a stable emulsion is formed. The detection of the composition of fatty acids in the in vitro digestion products shows that after the anthocyanin preparation containing the milk fat globule is digested in the simulated gastrointestinal fluid for 120 min, it is rich in long-chain unsaturated fatty acids such as tetradecanoic acid (1.27 μg / mL), cis-9-tetradecenoic acid (1.09 μg / mL), cis-9-hexadecenoic acid (1.98 μg / mL), cis-10-heptadecenoic acid (9.77 μg / mL), trans-9-octadecenoic acid (10.13 μg / mL), cis-9-octadecenoic acid (8.01 μg / mL), and medium-chain fatty acids such as octanoic acid (1.17 μg / mL), decanoic acid (1.64 μg / mL), undecanoic acid (0.98 μg / mL) and dodecanoic acid (1.03 μg / mL), and the ratio of medium-chain fatty acids to long-chain fatty acids is 1:8.

[0030] Example 2 Fresh raw milk was collected, impurities were removed, the pH was adjusted to 4.5, and the upper cream phase was collected by centrifugation at 5000 rpm at 4 °C, and mixed with sterile water and centrifuged at 5000 rpm to collect the fat layer, then washed with deionized water to collect the oil phase to remove protein, and repeated washing 3 times to obtain milk fat globules. 100 g of deionized water was taken, the pH was adjusted to 5.0, 4 g of milk fat globules and 0.15 g of black medlar anthocyanins were added and mixed, and then emulsified by first ultrasonic treatment (30%, 5 s on / 3 s off) for 5 min, and then shearing treatment (10000 rpm) for 10 min using a high-speed shearing machine to obtain an emulsion with a particle size of 256 nm.

[0031] The multiple interactions of electrostatic interaction, hydrogen bond interaction, van der Waals force and hydrophobic interaction between the black nightshade Lycium ruthenicum Murr. anthocyanins and the lipid membrane of milk fat globules were detected by fluorescence spectroscopy, Fourier infrared spectroscopy and isothermal titration calorimetry. The black nightshade Lycium ruthenicum Murr. anthocyanins were adsorbed on the interface of the milk fat globule membrane, the acylated group was inserted into the hydrophobic part of the lipid membrane to increase the hydrophobicity and membrane fluidity of the lipid membrane, and a stable emulsion was formed. The composition of fatty acids in the in vitro digestion product was detected. After the anthocyanin preparation containing milk fat globules was digested in the simulated gastrointestinal fluid for 120 min, it was rich in cis-9-tetradecenoic acid (2.72 μg / mL), hexadecanoic acid (5.90 μg / mL), cis-9-hexadecenoic acid (12.32 μg / mL), octadecanoic acid (7.11 μg / mL), trans-9-octadecenoic acid (8.56 μg / mL), cis-9-octadecenoic acid (1.42 μg / mL), cis-4,7,10,13,16,19-docosahexaenoic acid (10.62 μg / mL) and other long-chain unsaturated fatty acids, and contained hexanoic acid (1.81 μg / mL), octanoic acid (1.45 μg / mL), decanoic acid (2.16 μg / mL) and other medium-chain fatty acids, and the ratio of medium-chain fatty acids to long-chain fatty acids was about 1:9.

[0032] Example 3 Fresh raw milk was removed of impurities, the pH was adjusted to 4.5, and the upper cream phase was collected by centrifugation at 4 °C and 5000 rpm, and mixed with sterile water and centrifuged at 5000 rpm to collect the fat layer, and then washed with deionized water to collect the oil phase to remove proteins, and the washing was repeated 3 times to obtain milk fat globules. 100 g of deionized water was taken, the pH was adjusted to 4.5, 8 g of milk fat globules and 0.45 g of black nightshade Lycium ruthenicum Murr. anthocyanins were added and mixed, and then emulsified by first ultrasonic treatment (30%, 5 s on / 3 s off) for 5 min and then shearing treatment (12000 rpm) for 10 min using a high-speed shearing machine to obtain an emulsion with a particle size of 205 nm.

[0033] The multiple interactions of static interaction, hydrogen bond, van der Waals force and hydrophobic interaction between the black nightshade Lycium ruthenicum Murr. anthocyanins and the fat membrane of milk fat globule were detected by fluorescence spectroscopy, Fourier infrared spectroscopy and isothermal titration calorimetry. The black nightshade Lycium ruthenicum Murr. anthocyanins were adsorbed on the interface of the milk fat globule membrane, the acylated group was inserted into the hydrophobic part of the fat membrane to increase the hydrophobicity and membrane fluidity of the fat membrane, and a stable emulsion was formed. The composition of the fatty acids in the in vitro digestion product was detected. After the anthocyanin preparation containing milk fat globules was digested in the simulated gastrointestinal fluid for 120 min, it was rich in long-chain unsaturated fatty acids such as tetradecanoic acid (0.98 μg / mL), cis-9-tetradecenoic acid (1.25 μg / mL), cis-9-hexadecenoic acid (4.34 μg / mL), trans-9-octadecenoic acid (2.02 μg / mL), cis-9-octadecenoic acid (6.78 μg / mL), cis-4,7,10,13,16,19-docosahexaenoic acid (8.39 μg / mL), and contained medium-chain fatty acids such as hexanoic acid (1.27 μg / mL), octanoic acid (1.41 μg / mL), decanoic acid (0.75 μg / mL) and dodecanoic acid (0.86 μg / mL), and the ratio of medium-chain fatty acids to long-chain fatty acids was about 1:7.

[0034] Example 4 Fresh raw milk was removed impurities, pH was adjusted to 4.5, and the upper cream phase was collected by centrifugation at 5000 rpm at 4 °C, and mixed with sterile water and centrifuged at 5000 rpm to collect the fat layer, then washed with deionized water to collect the oil phase to remove protein, and repeated washing 3 times to obtain milk fat globules. 100 g of deionized water was taken, the pH was adjusted to 4.5, 6.5 g of milk fat globules and 0.3 g of black nightshade Lycium ruthenicum Murr. anthocyanins were added and mixed, and then emulsified by first ultrasonic treatment (30%, 5s on / 3s off) for 3 min, and then shearing treatment (10000 rpm) for 10 min by a high-speed shearing machine to obtain an emulsion with a particle size of 200 nm.

[0035] The multiple interactions of static interaction, hydrogen bond interaction, van der Waals force and hydrophobic interaction between the black wolfberry anthocyanin and the lipid membrane of the milk fat globule were detected by fluorescence spectroscopy, Fourier infrared spectroscopy and isothermal titration calorimetry, and the black wolfberry anthocyanin was adsorbed on the interface of the milk fat globule membrane, the acylated group was inserted into the hydrophobic part of the lipid membrane to increase the hydrophobicity and membrane fluidity of the lipid membrane, and a stable emulsion was formed. The composition of the fatty acid in the in vitro digestion product was detected, and it was found that after the anthocyanin preparation containing the milk fat globule was digested in the simulated gastrointestinal fluid for 120 min, it was rich in long-chain unsaturated fatty acids such as tetradecanoic acid (0.89 μg / mL), cis-9-tetradecenoic acid (1.02 μg / mL), hexadecanoic acid (2.23 μg / mL), cis-9-hexadecenoic acid (1.68 μg / mL), octadecanoic acid (4.32 μg / mL), trans-9-octadecenoic acid (2.13 μg / mL), cis-9-octadecenoic acid (7.01 μg / mL), cis-4,7,10,13,16,19-docosahexaenoic acid (7.99 μg / mL), medium-chain fatty acids such as hexanoic acid (0.98 μg / mL), octanoic acid (1.65 μg / mL) and decanoic acid (1.44 μg / mL), and the ratio of medium-chain fatty acids to long-chain fatty acids was about 1:7.

[0036] Comparative Example: Since the main components of the milk fat globule membrane are phospholipids and membrane proteins, accounting for 0.2% to 1.0% of the total fat, which is similar to the structure of liposomes. Therefore, in Comparative Examples 1 to 3, anthocyanin liposomes and anthocyanin milk fat globule membranes were respectively constructed.

[0037] Comparative Example 1 The black wolfberry anthocyanin liposomes were simulated to construct, and the structure characteristics, stability and digestion characteristics of the anthocyanin liposomes were evaluated and compared according to the technical idea. According to the method of (CN201210177952.1), soybean phospholipid and cholesterol were dissolved in 20 mL of anhydrous ethanol at a mass ratio of 1:1, and then 10 mL of an aqueous solution of black wolfberry anthocyanin extract was added and stirred uniformly. Ultrasonic-assisted reverse phase vacuum rotary evaporation was performed at an ultrasonic power of 180 W, a vacuum degree of 0.8 MPa, a rotation speed of 70 r / min, a water bath temperature of 30°C and a time of 25 min to obtain a liposome suspension. The liposome suspension was completely hydrated at room temperature for 1.5 h, filtered through a filter membrane with a pore size of 0.45 μm, and then black wolfberry anthocyanin liposomes were obtained. The particle size was 1.24 μm and the polydispersity index PDI was 0.56 (greater than 0.3, poor system stability). On this basis, the liposomes were prepared by the ultrasonic-assisted homogenization method of the present technology, and the particle size was 190 nm and the PDI was 0.22. Therefore, the anthocyanin liposome prepared by the technical solution has a nano-level structure, better dispersibility and stability, and is suitable for a wide range of food processing scenarios.

[0038] Comparative Example 2 The MFGM was prepared by using sphingomyelin, dilinoleoylphosphatidylethanolamine, dipalmitoylphosphatidylcholine, phosphatidylserine, phosphatidylinositol and cholesterol as materials. All components were dissolved in chloroform, vortexed to form a uniform lipid solution, dried in a fume hood or mild nitrogen stream, then hydrated with PBS (1x, pH 7.0) buffer to a lipid concentration of 0.5 mg / mL, 55°C water bath and vortexed for 5-8 min until the lipid membrane was fully hydrated and detached, probe sonication (300 W, 5s on / 3s off) for 10 min until the solution was translucent, and 0.5 mg / mL MFGM was prepared. Black wolfberry anthocyanins (pH 3.0, 3 mmol / L) were added at a molar ratio of 6:4, probe sonication (300 W, 5s on / 3s off) for 10 min, homogenized at 9000 rpm for 3 min, and anthocyanin-MFGM emulsion was prepared. The particle size was 425 nm and PDI=0.26; fluorescence spectroscopy, Fourier infrared spectroscopy, isothermal titration calorimetry were used to detect the electrostatic interaction, hydrogen bond interaction and hydrophobic interaction between black wolfberry anthocyanins and MFGM, and the structure was spherical (vesicle), and the black wolfberry anthocyanins were encapsulated in the hydrophilic part of the MFGM. No medium-chain fatty acids were detected after the system was digested in vitro for 120 min. The anthocyanin preparation prepared by using extracted milk fat globules as carriers in this technical solution is rich in medium-chain and long-chain unsaturated fatty acids, which is better than the MFGM prepared by various phospholipids, cholesterol and other exogenous materials.

[0039] Comparative Example 3 According to the technical solution, the preparation and property evaluation of single milk fat globule membrane (without black wolfberry anthocyanins) are implemented. Fresh raw milk (remove impurities) is centrifuged at 4 ℃ and 5000 rpm to collect the upper cream phase, mixed with sterile water and centrifuged at 5000 rpm to collect the fat layer, then washed with deionized water to collect the oil phase to remove protein, and repeated washing 3 times to obtain milk fat globule extract. First, the probe ultrasonic instrument (30%, 5s on / 3s off) is used for ultrasonic treatment for 5 min, and then the high-speed shearing machine 9000 rpm is used for shearing treatment for 10 min for emulsification, to obtain an emulsion with a particle size of 650 nm. After the emulsion containing milk fat globules is digested in vitro for 120 min, it contains hexanoic acid (2.36 μg / mL), decanoic acid (1.74 μg / mL), hexadecanoic acid (6.34 μg / mL), cis-9-octadecenoic acid (1.27 μg / mL), trans, trans-9,12-octadecadienoic acid (1.9 μg / mL), cis, cis, cis-9,12,15-octadecatrienoic acid (1.16 μg / mL), cis-4,7,10,13,16,19-docosahexaenoic acid (0.624 μg / mL) and other long-chain unsaturated fatty acids. Compared with examples 1 or 2 or 3, the types and contents of fatty acids are reduced. This result further confirms the beneficial effect of black wolfberry anthocyanins in the present application on the release of fatty acids.

[0040] In summary, the anthocyanin ingredient containing milk fat globules of the present application is a new type of nano-sized stable emulsion with natural milk fat globules as the anthocyanin carrier. The characteristic black wolfberry anthocyanins are adsorbed on the interface of the milk fat globule membrane, and the acylated groups are inserted into the hydrophobic part of the lipid membrane to form a stable structure, which has the effect of synergistic enzyme promoting the release of fatty acids. After the ingredient is digested in vitro, it contains rich long-chain unsaturated fatty acids and medium-chain fatty acids with a specific ratio, and is particularly suitable as a nutrient and healthy food ingredient, with a wide range of applications. The preparation method of the anthocyanin ingredient containing milk fat globules adopts the probe ultrasonic combined high-speed shearing technology to obtain an emulsion with better dispersibility and stability, and is simple, efficient, green and safe.

Claims

1. A anthocyanin ingredient comprising milk fat globules, characterized in that, It comprises milk fat globules, anthocyanins and water, and contains 4-10 g of milk fat globules and 0.15-0.5 g of anthocyanins per 100 g of water.

2. A berry ingredient comprising milk fat globules according to claim 1, characterized in that, The milk fat globules are separated from fresh milk, and the anthocyanins are obtained by extraction from Lycium ruthenicum Murr from Toksun, Xinjiang, and mainly comprise petunidin-3-O-rutinoside (coumaroyl) -5-O-glucoside with a content of greater than or equal to 95% and a small amount of cyanidin-3-O-coumaroyl-glucoside.

3. The anthocyanin ingredient containing milk fat globules of claim 1, wherein, The milk fat globule-containing anthocyanin preparation is a stable emulsion with a specific structure formed by multiple interactions of electrostatic interaction, hydrogen bonding, van der Waals force and hydrophobic interaction between the fat membrane of the milk fat globules and the anthocyanins from Lycium ruthenicum Murr, and has a particle size of less than 980 nm.

4. The anthocyanin ingredient containing milk fat globules of claim 1, wherein, The milk fat globule-containing anthocyanin preparation is rich in long-chain unsaturated fatty acids and medium-chain fatty acids with a specific ratio of 1:7-10 after in-vitro digestion in simulated gastrointestinal fluid.

5. A process for the preparation of a anthocyanin ingredient containing milk fat globules according to claims 1 to 4, characterized in that It comprises the following preparation steps: (1) extracting milk fat globules from fresh raw milk; (2) adding anthocyanins and milk fat globules to deionized water for emulsification to obtain an emulsion, i.e., a milk fat globule-containing anthocyanin preparation; In step (1), the milk fat globules are obtained by taking fresh raw milk, removing impurities, adjusting the pH to 4.5, centrifuging the upper cream phase at 4 °C at 3000-8000 rpm, mixing with sterile water, centrifuging the fat layer at 3000-8000 rpm, washing the oil phase with deionized water to remove proteins, repeating the washing 2-3 times, and obtaining a paste extract, i.e., the milk fat globules. In step (2), the Lycium ruthenicum Murr anthocyanins in claim 2 and the milk fat globules in step (1) are added to deionized water for emulsification, wherein the milk fat globules are added in a mass fraction of 4-10%, the anthocyanins are added in a mass fraction of 0.15-0.5%, and the deionized water has a pH of 4.0-5.

0.

6. The production method according to claim 5, wherein In step (2), the emulsification is first treated with a probe-type ultrasonic instrument (30%, 5 s on / 3 s off) for 2-5 min, and then treated with a high-speed shearing machine at 9000-12000 rpm for 3-10 min, and the emulsion is balanced for 24 h without delamination, thereby obtaining an emulsion with a particle size of less than 980 nm, i.e., the milk fat globule-containing anthocyanin preparation.