A high-energy fluid mill-mediated embedding composite system and its application in functional food

Abstract

The application discloses a high-energy fluid mill-mediated embedding composite system and application thereof in functional food, and relates to the technical field of food processing and natural product homeostasis. The embedding composite system is prepared by the following steps: wet pre-pulverizing fruits and vegetables and a natural matrix with embedding function, and adding the fruits and vegetables and the natural matrix into a high-energy fluid mill for treatment; and the embedding composite system is treated by wet pulverization and ultrahigh-pressure jetting, so that polysaccharides and other active ingredients in the natural matrix and anthocyanins and polyphenols in the fruits and vegetables form a stable embedding structure under the action of mechanical force. The preparation method is simple and efficient, does not need to add chemical color protectants and stabilizers, is suitable for industrial production, and the obtained embedding composite system has excellent color stability, high active ingredient retention rate and good storage performance, effectively delays browning and precipitation, realizes natural and clean label quality improvement of fruit and vegetable juice, and provides an innovative solution for color protection and stabilization processing of blueberries and other fruit and vegetable juices.

Description

Technical Field

[0001] This invention relates to the field of functional food processing and natural product stabilization technology, specifically to a high-energy fluid mill-mediated encapsulation composite system and its application in functional foods. Background Technology

[0002] Blueberries are rich in anthocyanins, polyphenols, and other bioactive components, and have attracted much attention in the functional food and beverage industry due to their excellent antioxidant, anti-inflammatory, and metabolic regulation functions. Studies have shown that blueberry anthocyanins have significant antioxidant capacity, but their poor physicochemical stability makes them susceptible to degradation and browning under the influence of light, heat, oxygen, and enzymes. This leads to color deterioration, loss of active ingredients, and bioavailability often below 10%, severely limiting their long-term efficacy in industrialized beverage products. Traditional blueberry juice processing relies on chemical color-protecting agents and synthetic stabilizers, which raise safety concerns and do not conform to the trend of clean labeling. Physical color-protecting methods, such as high-temperature instantaneous sterilization, accelerate the destruction of heat-sensitive components, and conventional high-pressure homogenization or colloid milling processes are insufficient to achieve stable binding of active ingredients and carrier materials, resulting in problems such as short-lasting color protection and precipitation.

[0003] Chinese patent CN202410226933.6 discloses a method for preparing blueberry anthocyanin microcapsules. The core material is obtained through extraction and purification, then mixed with carboxymethyl starch-gellan gum wall material, and freeze-dried to form microcapsules. While this method can improve anthocyanin stability, it suffers from drawbacks such as complex processes, encapsulation efficiency being affected by multiple factors, and high costs for large-scale production. Chinese patent CN201610579635.0 describes a method for preparing a blueberry juice stabilizer using ethyl acetate extraction of green tea extract containing NaHCO3. This stabilizer can improve anthocyanin stability and antioxidant capacity, but research on its long-term stabilization protection mechanism is insufficient. Chinese patent CN202310508197.9 describes an intramolecular co-coloring method using cobalt source irradiation to treat freshly squeezed blueberry juice. This method is simple, green, and safe, and can improve color stability and shelf life. However, this method relies on external irradiation and does not construct an internal stabilization encapsulation structure, resulting in significant fluctuations in the retention rate of active ingredients during long-term storage. Chinese patent CN202311613870.1 describes a method that uses papain combined with high-pressure carbon dioxide sterilization to treat mulberry juice, effectively inhibiting browning and discoloration and preserving active ingredients and flavor during long-term storage. However, this method relies on specific sterilization conditions, does not introduce molecular-level structural stabilization strategies, and lacks in-depth explanation of the long-term protective mechanisms for components such as anthocyanins. Summary of the Invention

[0004] The purpose of this invention is to at least solve one of the technical problems existing in the prior art, and to provide a high-energy fluid mill-mediated encapsulation composite system and its application in functional foods. Specifically, it relates to a blueberry polyphenol-tremella polysaccharide encapsulation composite system based on high-energy fluid mill ultra-high pressure jet treatment and its application in the color protection and stabilization of blueberry juice. In particular, it relates to a technical solution for achieving efficient protection and long-term stability of heat-sensitive active ingredients such as blueberry anthocyanins through mechanical force-mediated molecular encapsulation technology.

[0005] The technical solution of the present invention is as follows: The first aspect of the present invention provides a method for preparing a high-energy fluid mill-mediated embedded composite system, comprising the following steps: S1. After cleaning the fresh fruits and vegetables and the natural matrix with the embedding function, add pure water and perform wet pre-pulverization to obtain a preliminary homogeneous mixed slurry. S2. The mixed slurry is added to a high-energy fluid mill for ultrafine grinding. Under the high shear, cavitation and impact of the high-energy fluid mill, the natural matrix with encapsulation function performs in-situ molecular-level composite and encapsulation of the active ingredients in the fresh fruits and vegetables to prepare the encapsulation composite system. The active ingredients in the fresh fruits and vegetables are at least one of polyphenols, anthocyanins, vitamins, polysaccharides, organic acids, and terpenoids; the natural matrix with encapsulation function is at least one of a natural matrix containing polysaccharides, a natural matrix containing organic acids, and a natural matrix containing terpenoids.

[0006] This invention innovatively employs high-energy fluid milling and ultra-high-pressure jet technology, combined with a natural matrix possessing encapsulation capabilities, to construct a composite system for encapsulating active ingredients in fruits and vegetables. Through the extreme shearing, cavitation, and impact effects of the high-energy fluid milling, this invention forces components such as polysaccharides in the natural matrix and active ingredients like anthocyanins, polyphenols, and cyanidins in fruits and vegetables to form a dense and uniform encapsulation structure at the molecular level. This achieves long-lasting color protection, physical stability, and functional retention of fruits and vegetables without the need for chemical additives. Compared to existing technologies, this invention offers a simple and efficient process suitable for continuous production, solving key problems such as low encapsulation rates, poor stability, and complex processes in traditional methods. It provides an innovative solution for the clean labeling and high-value development of functional foods or beverages such as blueberry juice and other fruit and vegetable juices.

[0007] Optionally, the fresh fruits and vegetables are at least one of blueberries, grapes, mulberries, strawberries, mangoes, pomegranates, broccoli, tomatoes, and pumpkins; The natural matrix with encapsulation function is at least one of the following: Tremella fuciformis, bamboo fungus, Poria cocos, inulin, and sodium alginate.

[0008] The following amounts of each ingredient are added by weight: 20-40 parts fresh fruits and vegetables, 1-4 parts natural matrix with encapsulation function, and 40-80 parts purified water.

[0009] Optionally, in S1, a wet pulverizing device is used for wet pre-pulverization. The wet pre-pulverization process operates at a speed of 2500~3500 rpm, using 18~22° hammers and a toothed ring with 0 cutting depth. The processing time for the wet pre-pulverization is 10 min~20 min. More preferably, the wet pre-pulverization process operates at a speed of 3000 rpm, using 20° hammers and a toothed ring with 0 cutting depth. The processing time for the wet pre-pulverization is 15 min.

[0010] 18~22° hammerheads refer to hammerheads with a working face angle of 18~22°. Compared to sharper angles (such as 10°-15°), 18~22° hammerheads are blunter and thicker, focusing more on impact force than cutting force. Zero cutting depth means the gap between the moving blade (hammerhead) and the fixed blade (or screen, bottom blade) is close to zero, or the depth to which the blade teeth cut into the material is set to the minimum. In industrial crushing, "zero cutting depth" often means the hammerhead and screen or bottom blade are in critical contact, or the equipment mainly relies on extrusion and friction to crush materials, rather than relying on the blade tip "biting" into the material for cutting. This invention uses 18~22° hammerheads and a toothed ring with zero cutting depth to crush, knead, and compress the raw materials until they are broken, thereby obtaining a uniform and fine mixed slurry.

[0011] In S2, the jet pressure of the high-energy fluid mill is 30-280 MPa. Preferably, the jet pressure of the high-energy fluid mill is 80-150 MPa; more preferably, the jet pressure of the high-energy fluid mill is 120 MPa.

[0012] The high-energy fluid mill in this invention provides high shear, cavitation, and impact forces to force the components in the natural matrix, such as polysaccharides, and active ingredients like anthocyanins, polyphenols, and cyanidins from fruits and vegetables, into a dense and uniform encapsulation structure at the molecular level. Therefore, the jet pressure of the high-energy fluid mill is crucial. Research has shown that if the jet pressure is too low, the intensity of high shear, cavitation, and impact forces is insufficient, making it impossible to form a dense and uniform encapsulation structure. This results in poor dispersion of active ingredients and easy leakage. Simultaneously, the failure of the cavitation effect leads to low encapsulation efficiency and poor process economy. Conversely, if the jet pressure is too high, it easily causes oxidative degradation of active ingredients such as anthocyanins, polyphenols, and cyanidins, and the molecular chains of the natural matrix break, destroying the encapsulation capacity. It can also cause problems such as localized overheating and increased equipment wear. Therefore, this invention preferably uses a pressure of 120 MPa.

[0013] A second aspect of the present invention provides an encapsulation complex system obtained by the preparation method described above, the encapsulation complex system comprising a natural matrix with encapsulation function and fresh fruits and vegetables, wherein the natural matrix with encapsulation function encapsulates the active ingredients in the fresh fruits and vegetables through intermolecular forces to form a stable complex or microcapsule structure.

[0014] Optionally, the particle size of the encapsulated composite system is as follows: D[3,2] is 13~15 μm, D[4,3] is 34~36 μm, D10 is 5~7 μm, D50 is 26~28 μm, and D90 is 75~78 μm.

[0015] A third aspect of the present invention provides an application of the aforementioned encapsulation composite system in the preparation of functional foods.

[0016] A fourth aspect of the present invention provides a functional food beverage, characterized in that the functional food beverage is obtained by sterilization of the encapsulation complex system.

[0017] Optionally, the sterilization method is ultra-high temperature instantaneous sterilization, and the conditions for ultra-high temperature instantaneous sterilization are 135–140°C for 3–5 seconds.

[0018] Optionally, the functional food and beverage is a compound blueberry juice, wherein the fresh fruit and vegetable is blueberry, and the natural matrix with encapsulation function is tremella; the compound blueberry juice has an anthocyanin retention rate of not less than 85% after accelerated storage test, and a browning index of not more than 2.5 after 30 days of storage.

[0019] This invention innovatively employs high-energy fluid milling and ultra-high pressure jet technology, combined with natural Tremella fuciformis polysaccharide as a carrier, to construct a blueberry active ingredient encapsulation complex system. This technology, through extreme shearing, cavitation, and impact, forces Tremella fuciformis polysaccharide to form a dense and uniform encapsulation structure with blueberry anthocyanins and polyphenols at the molecular level, achieving long-lasting color protection, physical stability, and functional retention of blueberry juice without the need for chemical additives.

[0020] Optionally, the compound blueberry juice can be consumed alone or used as a base in products such as fruit juice beverages, jams, and functional foods.

[0021] Optionally, the preparation method of the compound blueberry juice includes the following steps: Step 1: After cleaning the fresh blueberries and white fungus according to the mass ratio, add an appropriate amount of purified water and use a wet crushing equipment for pre-crushing treatment to obtain a preliminary homogeneous mixed slurry. Step 2: The above mixed slurry is fed into a high-energy fluid mill system. During this process, the tremella polysaccharide and blueberry active ingredients will further achieve in-situ molecular-level compounding and encapsulation under high shear, cavitation and impact to form a full-component compound blueberry juice. Step 3: The whole-component blueberry juice obtained in Step 2 is subjected to ultra-high temperature (UHT) sterilization to achieve microbial control while preserving the active ingredients to the maximum extent; Step 4: Use the sterilized compound blueberry juice directly as the final product, or blend and bottle it as needed.

[0022] The fifth aspect of the present invention provides an application of the preparation method described above in the color protection and stabilization processing of fruit and vegetable juices.

[0023] This invention involves directly wet pre-pulverizing fresh fruits and vegetables with natural matrices such as tremella, followed by ultrafine pulverization using a high-energy fluid mill. This process allows the polysaccharides from tremella to be in situ encapsulated and structurally integrated with the polyphenols, anthocyanins, and other active ingredients in the fresh fruits and vegetables under mechanical force, forming a fully-component composite system with excellent stability. This process requires no addition of any chemically synthesized color-protecting agents or stabilizers, and can significantly improve the stability, active ingredient retention rate, and shelf life of the compound blueberry juice while retaining all the nutrients of the raw materials. Therefore, the above method can be applied to the color protection and stabilization processing of other fruit and vegetable juices.

[0024] Optionally, the above method can be used in the preparation of blueberry and tremella encapsulation complex system and complex blueberry juice containing the system. The complex blueberry juice prepared by the above method has an anthocyanin retention rate of no less than 85% after accelerated storage test and a browning index of no more than 2.5 after 30 days of storage, showing excellent color protection effect and physical stability.

[0025] This invention has at least one of the following beneficial effects: 1. This invention innovatively employs high-energy fluid mill ultra-high pressure jet technology, combined with a natural matrix with encapsulation function as a carrier, to construct a composite system for encapsulating active ingredients in fruits and vegetables. Through the extreme shearing, cavitation, and impact effects of the high-energy fluid mill, the components in the natural matrix, such as polysaccharides, and active ingredients such as anthocyanins, polyphenols, and anthocyanins in fruits and vegetables are forcibly encapsulated at the molecular level to form a dense and uniform encapsulation structure, avoiding the loss of active ingredients in fruits and vegetables and preserving the natural nutrition of fruits and vegetables and the natural matrix with encapsulation function to the greatest extent.

[0026] 2. This invention utilizes wet pulverization and high-energy fluid milling to achieve in-situ encapsulation of polysaccharides and other components in natural matrices with active ingredients in fruits and vegetables, without the need for chemical stabilizers. The resulting compound fruit and vegetable juice exhibits excellent color stability, high retention rate of active ingredients, and good storage performance. The process of this invention is simple, highly continuous, and suitable for industrial production, possessing high value for promotion and application. Attached Figure Description

[0027] Figure 1 This is a flowchart illustrating the preparation process of the full-component compound blueberry juice in Example 1 of the present invention; Figure 2 This is a schematic diagram of the high-energy fluid mill in Embodiment 1 of the present invention; Figure 3 The images show a comparative scanning electron microscope (SEM) image of the microstructure of the blueberry-tremella polysaccharide encapsulation complex system prepared in Example 1 and Comparative Example 1, respectively; wherein, A in the image corresponds to Comparative Example 1 and B in the image corresponds to Example 1. Figure 4 The particle size distribution and zeta potential diagrams of Example 1 and Comparative Examples 1-3 of the present invention are shown. Figure 5 The color difference between Example 1 and Comparative Examples 1-3 of this invention; Figure 6 The figures show the apparent stability, settling rate, and accelerated storage test data of Example 1 and Comparative Examples 1-3 of the present invention. In the figures, A represents apparent stability, with the left bottle corresponding to Comparative Example 1 and the right bottle corresponding to Example 1; B represents settling rate and accelerated storage test data. Figure 7 The Fourier transform spectra of Embodiment 1 and Comparative Example 1 of the present invention are shown below. Figure 8 These are the XRD patterns of Embodiment 1 and Comparative Example 1 of the present invention; Figure 9 Thermogravimetric analysis diagrams of Embodiment 1 and Comparative Example 1 of the present invention; Figure 10 The graph shows the polyphenol oxidase activity assay data for Example 1 and Comparative Example 1 of this invention. Figure 11 This is a graph showing the anthocyanin retention rates of Example 1 and Comparative Example 1 of the present invention. Detailed Implementation

[0028] To make the technical problems solved, the technical solutions, and the beneficial effects of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

[0029] The wet pulverizing equipment used in the following examples and comparative examples was purchased from Wuxi Xinhai Drying Powder Equipment Co., Ltd. The schematic diagram of the high-energy fluid mill used in the following embodiments and comparative examples is shown below. Figure 2 As shown, the specific structure of the high-energy fluid mill can be found in Chinese Patent CN 119158674 A.

[0030] Example 1 A method for preparing a high-energy fluid mill-mediated in-situ encapsulation complex of blueberry and tremella fuciformis and its application in whole-component blueberry juice. Figure 1The diagram shows the process flow for preparing full-component compound blueberry juice, including the following steps: Step 1: Select 20 kg of fresh blueberries and 1 kg of white fungus, wash and set aside; Step 2: Mix blueberries and white fungus, add 40 kg of purified water, and pre-crush in a wet grinding equipment (3000 rpm, 20° hammer, 0 cutting depth toothed ring) for 15 min; Step 3: Feed the pre-crushed slurry into a high-energy fluid mill and process it under a pressure of 120 MPa; Step 4: The treated pulp is subjected to ultra-high temperature instantaneous sterilization (135℃, 4 s), cooled and then bottled to obtain compound blueberry juice (CMJ-120). Step 5: Perform performance testing on the obtained compound blueberry juice.

[0031] Example 2 A method for preparing a high-energy fluid mill-mediated in-situ encapsulation complex of blueberry and tremella fuciformis and its application in whole-component blueberry juice. Compared with Example 1, the processing pressure of the high-energy fluid mill was changed to 60 MPa. The specific preparation method is as follows: A blueberry-tremella in situ encapsulation complex system under different pressures includes the following steps: Step 1: Select 20 kg of fresh blueberries and 1 kg of white fungus, wash and set aside; Step 2: Mix blueberries and white fungus, add 40 kg of purified water, and pre-grind in a wet grinding equipment for 15 min; Step 3: Feed the pre-crushed slurry into a high-energy fluid mill and process it under a pressure of 60 MPa; Step 4: The treated pulp is subjected to ultra-high temperature instantaneous sterilization (135℃, 4 s), cooled and then bottled to obtain blueberry juice (CMJ-60). Step 5: Perform performance testing on the obtained compound blueberry juice.

[0032] Example 3 A method for preparing a high-energy fluid mill-mediated in-situ encapsulation composite system of mulberry and tremella and its application in whole-component mulberry juice. Compared with Example 1, the raw material is changed from fresh blueberries to fresh mulberries, and the other steps are the same as in Example 1.

[0033] Example 4 A method for preparing a tomato-Tremella fuciformis in-situ encapsulation composite system mediated by a high-energy fluid mill and its application in whole-component tomato juice. Compared with Example 1, the raw material fresh blueberries are replaced with fresh tomatoes, and the other steps are the same as in Example 1.

[0034] Comparative Example 1 Compared with Example 1, no tremella was added. The specific preparation method is as follows: A method for preparing whole-component blueberry juice without added tremella includes the following steps: Step 1: Select 20 kg of fresh blueberries, wash and set aside; Step 2: Mix 20 kg of blueberries with 40 kg of purified water and pre-grind them in a wet grinding equipment for 15 min; Step 3: Feed the pre-crushed slurry into a high-energy fluid mill and process it under a pressure of 120 MPa; Step 4: The treated pulp is subjected to ultra-high temperature instantaneous sterilization (135℃, 4 s), cooled and then bottled to obtain blueberry juice (abbreviated as BJ-120). Step 5: Perform performance testing on the obtained blueberry juice.

[0035] Comparative Example 2 Compared to Example 1, no tremella was added and the processing pressure of the high-energy fluid mill was changed to 0 MPa. The specific preparation method is as follows: A method for preparing whole-component blueberry juice without added tremella and without pressure includes the following steps: Step 1: Select 20 kg of fresh blueberries, wash and set aside; Step 2: Mix 20 kg of blueberries with 40 kg of purified water and pre-grind them in a wet grinding equipment for 15 min; Step 3: Feed the pre-crushed slurry into a high-energy fluid mill and process it under 0 MPa pressure; Step 4: The treated pulp is subjected to ultra-high temperature instantaneous sterilization (135℃, 4 s), cooled and then bottled to obtain blueberry juice (abbreviated as BJ-0). Comparative Example 3 Compared with Example 1, the processing pressure of the high-energy fluid mill was changed to 0 MPa, and the specific preparation method is as follows: A stress-free blueberry-tremella in-situ encapsulation complex system includes the following steps: Step 1: Select 20 kg of fresh blueberries and 1 kg of white fungus, wash and set aside; Step 2: Mix blueberries and white fungus, add 40 kg of purified water, and pre-grind in a wet grinding equipment for 15 min; Step 3: Feed the pre-crushed slurry into a high-energy fluid mill and process it under 0 MPa pressure; Step 4: The treated pulp is subjected to ultra-high temperature instantaneous sterilization (135℃, 4 s), cooled and then bottled to obtain compound blueberry juice (CMJ-0). Step 5: Perform performance testing on the obtained compound blueberry juice.

[0036] Comparative Example 4 Compared to Example 1, step 3 uses a commercially available Joyoung P363 blender instead of a high-energy fluid mill, and sets the device to fruit and vegetable juice mode to process the raw materials. Everything else is the same as in Example 1, resulting in a compound blueberry juice, named CMJ-P.

[0037] The products prepared in the above embodiments and comparative examples were subjected to performance testing, and the testing methods are as follows: I. Detection Methods 1. Scanning Electron Microscopy (SEM): A suitable amount of sample is evenly coated onto a conductive adhesive, sputter-coated with gold, and then observed under a scanning electron microscope. Under a suitable accelerating voltage, the microstructure of the sample is scanned in multiple regions to obtain images of the particle surface structure and dispersion state, which are used to analyze the morphological characteristics of fruit pulp particles, cell debris, or additives.

[0038] Particle size: The particle size was determined using a laser particle size analyzer. After appropriate dilution, the sample was placed in a sample cell, and the particle size distribution was measured using the principle of laser scattering. The volume average particle size (D50) and polydispersity index (PDI) were recorded to reflect the particle size uniformity and suspension state of the solid particles in the juice.

[0039] Color difference: Using a colorimeter calibrated with a standard white plate, the sample is injected into a cuvette, and parameters such as lightness (L), red-green value (a), and yellow-blue value (b) are measured. By calculating the total color difference (ΔE) and saturation (C), the color change of the sample is quantified, and the color stability during processing or storage is evaluated.

[0040] Anthocyanin retention: The pH differential method was used. Samples were diluted in pH 1.0 and pH 4.5 buffer solutions, and absorbance was measured at 520 nm and 700 nm wavelengths to calculate anthocyanin content. Using fresh or initial samples as a baseline, the retention rate was calculated by comparing anthocyanin content at different processing stages to evaluate the degree of retention of active ingredients during processing.

[0041] Apparent stability: The sample is quantitatively loaded into a transparent graduated tube, sealed, and allowed to stand at room temperature or centrifuged at low speed. Changes in the layering interface height, precipitate volume, or supernatant transparency are observed and recorded periodically. The physical stability of the system is comprehensively evaluated by the precipitation rate or layering speed.

[0042] Sedimentation rate: The gravity sedimentation method was used to record the height of the solid-liquid interface as the sample was placed in the static state, and the sedimentation curve was plotted to calculate the sedimentation distance per unit time, thereby evaluating the sedimentation behavior and stability of the suspended particulate phase.

[0043] Fourier Transform Infrared Spectroscopy (FTIR): The freeze-dried powder of the sample was mixed with potassium bromide, compressed into tablets, and then spectrated at 4000-4000 cm⁻¹. -1 Infrared absorption spectra were obtained by scanning within the wavenumber range, and the changes in characteristic peaks of molecular functional groups were analyzed.

[0044] X-ray diffraction (XRD): The sample is freeze-dried into a solid powder and placed on the sample stage of an X-ray diffractometer. The diffraction pattern is obtained by scanning within a specific angular range (5°-80° 2θ), and the crystal structure of the sample is determined by the peak position and intensity, and its physical structural characteristics are analyzed.

[0045] Thermogravimetric analysis: The sample was freeze-dried into a solid powder and placed in the sample pan of the thermogravimetric analyzer. Under a nitrogen atmosphere, the temperature was increased at a constant rate of 10°C / min, and the mass change curve of the sample with temperature was recorded. The moisture, volatile components, and thermal stability were analyzed through the weight loss stage.

[0046] II. Test Results pass Figure 3 Scanning electron microscopy (SEM) images show that sample A (Comparative Example 1) exhibits a relatively loose structure with irregular particles and slight wrinkling on its surface. This is mainly due to the lack of a stable gel network to support the blueberry particles during the jetting process, leading to the collapse of its microstructure. In contrast, sample B (Example 1) displays significantly different structural characteristics. This is because the long-chain molecules of Tremella fuciformis polysaccharide are fully dissolved and dispersed, spontaneously constructing a continuous and dense three-dimensional gel network in the blueberry juice system. This network not only acts as a natural scaffold during the drying process, effectively preventing structural shrinkage and collapse, but more importantly, it can physically capture and fix functional components such as anthocyanins and polyphenols in the blueberry juice within its network, thereby achieving "in-situ encapsulation" of active ingredients.

[0047] Table 1 and Figure 4Particle size distribution analysis of the samples showed that the group treated with the Joyoung blender (Comparative Example 4) had smaller particle sizes, but excessive sedimentation, indicating not only very poor stability of the compound blueberry juice, which separated into layers after only a short time, but also a high proportion of large particles, resulting in a rough texture. The accumulation of fruit residue easily caused localized oxidative browning, leading to accelerated anthocyanin degradation. Due to excessive sedimentation, Comparative Example 4 was not further analyzed. In the groups without ultra-high pressure jet treatment, Comparative Example 2 had larger particle sizes and a relatively wider distribution, while the particle size distribution of Comparative Example 3 shifted to a smaller range, indicating that Tremella fuciformis polysaccharide may have a certain dispersing and encapsulating effect on the original blueberry juice particles through intermolecular interactions. After treatment with 120 MPa ultra-high pressure jet, the particle sizes of both samples (Comparative Example 1 and Example 1) were significantly reduced, and the distribution became more concentrated. The peak particle size distribution of Example 1 shifted further to the left, exhibiting a more obvious single-peak characteristic, with the vast majority of particles distributed below 10 μm. This indicates that ultra-high pressure jet treatment can effectively break down large particles in fruit juice and improve the uniformity of the system. The presence of Tremella polysaccharide may synergistically enhance the homogenization effect of jet treatment and further stabilize the fine particles through in-situ embedding mechanism, thereby forming a more stable and uniform composite system.

[0048] Table 1 Particle size distribution Note: In the figure, a, b, c, and d represent significant differences between groups. Different letters indicate significant differences (P<0.05), while the same letter indicates no significant differences.

[0049] from Figure 5 In terms of color difference indices, the 120 MPa jet treatment showed a consistent effect on the color of Example 1 and Comparative Example 1: whether it was single blueberry juice or compound blueberry juice, after jet treatment, the L value, representing brightness, decreased significantly, the a value, representing red-green hue, increased significantly, and the b value, representing yellow-blue hue, also increased. This indicates that the treated juice was darker and had a more pronounced red-yellow hue. Meanwhile, the L value of Example 1 was generally higher than that of single blueberry juice, indicating that the compound juice had a higher base brightness. The jet treatment had a similar trend in its effect on the color difference of the two types of juice, showing that the effect of this treatment on the color change of juice had a certain degree of universality.

[0050] from Figure 6From the perspective of the indicators, the 120 MPa jet treatment had a significant impact on the stability-related indicators of Example 1 and Comparative Example 1. The samples treated with jet treatment had significantly lower instability index, sedimentation rate, and solids content than the untreated samples, indicating that jet treatment can effectively improve the physical stability of the juice and reduce the risk of sedimentation and stratification of the system. At the same time, the overall stability indicators of the compound blueberry juice were higher than those of the single blueberry juice, indicating that the basic stability of the compound system was weaker than that of the single blueberry juice. However, combining the sedimentation rate and solids content, it can be found that the apparent stability of Example 1 was better.

[0051] pass Figure 7 Fourier transform infrared spectroscopy was used to analyze the structural differences between Example 1 and Comparative Example 1. The results showed that the overall configuration of the characteristic absorption peaks of the two were consistent, both around 3411 cm⁻¹. -1 A broad and strong OH stretching vibration peak exists at 1736 cm⁻¹. -1 and 1632 cm -1 The peaks at 1252 cm⁻¹ exhibited C=O stretching vibration and C=C skeletal vibration, respectively, indicating that the composite sample retained the major functional group structure of blueberry juice. However, compared to Comparative Example 1, Example 1 showed a peak at 1252 cm⁻¹. -1 The intensity of the CO stretching vibration absorption peak in the vicinity was significantly enhanced, and the peak shape and position of some characteristic peaks (such as OH and C=O) showed slight shifts and broadening. These changes are closely related to the enhanced hydrogen bonding caused by the abundant hydroxyl and carboxyl groups in Tremella fuciformis polysaccharide, indicating that a new intermolecular interaction force was formed between the active components of blueberry juice and Tremella fuciformis polysaccharide in the CMJ system. This confirms that Tremella fuciformis polysaccharide has a physical encapsulation effect on the blueberry juice components, further supporting the conclusion that an in-situ encapsulation structure is formed in the composite blueberry juice of Example 1.

[0052] pass Figure 8 XRD analysis revealed that both samples exhibited typical broad, diffuse peaks at 2θ≈20°, without sharp crystalline diffraction peaks, indicating that the overall sample structure was predominantly amorphous or low-crystallinity. Specifically, the diffraction peak intensity of Example 1 was significantly higher than that of Comparative Example 1, and the peak shape was fuller; this difference is the core basis for determining the existence of an embedded structure. The weak and flat peaks of Comparative Example 1 mainly reflect the amorphous characteristics of its components (such as polysaccharides and anthocyanins in blueberries) in their natural state. In contrast, the Tremella fuciformis polysaccharide introduced in Example 1, as a natural polymer with excellent film-forming and inclusion abilities, encapsulates the blueberry active ingredients through intermolecular forces such as hydrogen bonds and hydrophobic interactions, forming a stable complex or microcapsule structure. This inclusion effect induces rearrangement of the Tremella fuciformis polysaccharide molecular chains, forming a more regular secondary structure, which manifests as a significant increase in diffraction peak intensity and order in XRD. Therefore, Example 1 contains an embedded structure formed by Tremella fuciformis polysaccharide encapsulating the blueberry active ingredients.

[0053] pass Figure 9 Thermogravimetric analysis revealed that, within the main thermal decomposition range of 200–400℃, the thermal weight loss rate of Example 1 was significantly slower than that of Comparative Example 1, and the total weight loss rate was also lower at the same temperature. For example, at 400℃, the weight loss ratio of Example 1 was approximately 33%, while that of Comparative Example 1 was approximately 39%. This indicates that the structure of Example 1 is more stable and better resistant to thermal decomposition. This is because the Tremella fuciformis polysaccharide introduced in Example 1, acting as a wall material, encapsulates the active ingredients (such as anthocyanins and polyphenols) in blueberries through hydrogen bonds and hydrophobic interactions, forming a microcapsule-like encapsulation structure. This dense core-shell structure acts like a "protective shell," effectively delaying the thermal degradation and volatilization of the internal components, thereby improving the overall thermal stability, as reflected in the reduced weight loss rate on the TG curve. In contrast, the thermal weight loss curve of Comparative Example 1 is steeper, indicating that its components such as water, small molecule sugars, and free anthocyanins are more easily and rapidly decomposed and lost when heated, lacking the protection of a polymer wall material, thus resulting in poorer thermal stability.

[0054] Depend on Figure 10 It can be seen that, compared with Comparative Example 1, the contents of the three core active substances—anthocyanins, total phenols, and total flavonoids—in Example 1 all showed a significant upward trend. The anthocyanin content in Example 1 was slightly higher than that in Comparative Example 1, while the increases in the contents of total phenols and total flavonoids were particularly significant. The total phenol content increased substantially compared to Comparative Example 1, and the total flavonoid content also showed a significant advantage. This result fully demonstrates that the introduction of Tremella fuciformis does not simply dilute the active ingredients of blueberries, but rather forms a stable encapsulated complex structure through intermolecular hydrogen bonds and hydrophobic interactions. This plays a crucial role in protecting and stabilizing the anthocyanins, polyphenols, and flavonoids in blueberries, effectively inhibiting the oxidation, degradation, and loss of active ingredients during the preparation of the complex system. In contrast, Comparative Example 1, lacking the physical barrier and structural protection formed by Tremella fuciformis polysaccharides, was more prone to oxidative loss and molecular dissociation of active substances, ultimately resulting in a lower overall content of active substances. This phenomenon corroborates the conclusions of previous XRD and thermogravimetric analyses that Example 1 exhibited an ordered encapsulation structure and significantly better thermal stability than Comparative Example 1. This further confirms that the encapsulation effect of Tremella fuciformis polysaccharide on the active ingredients of blueberries is the core mechanism for improving the retention rate of active substances in the system, and also provides direct experimental evidence for the functional optimization of compound fruit juice.

[0055] Figure 11The results show that, during the 10-day storage period, the anthocyanin retention rate of Example 1 was consistently significantly higher than that of Comparative Example 1, and the degradation rate was more gradual. This result further confirms the stabilizing and protective effect of the Tremella fuciformis polysaccharide encapsulation structure on blueberry anthocyanins: the physical barrier formed by encapsulation and the intermolecular interactions effectively slowed down the oxidative degradation process of anthocyanins and improved their stability during storage. In contrast, Comparative Example 1, lacking structural protection, had anthocyanins that were more susceptible to degradation due to environmental factors, resulting in a consistently lower retention rate than Example 1. This is highly consistent with the previous XRD, thermogravimetric, and active substance content analyses, jointly confirming that Tremella fuciformis polysaccharide encapsulation is the core mechanism for improving the stability and retention rate of blueberry active substances.

[0056] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A method for preparing a high-energy fluid mill-mediated embedded composite system, characterized in that, Includes the following steps: S1. After cleaning the fresh fruits and vegetables and the natural matrix with the embedding function, add pure water and perform wet pre-pulverization to obtain a preliminary homogeneous mixed slurry. S2. The mixed slurry is added to a high-energy fluid mill for ultrafine grinding. Under the high shear, cavitation and impact of the high-energy fluid mill, the natural matrix with encapsulation function performs in-situ molecular-level composite and encapsulation of the active ingredients in the fresh fruits and vegetables to prepare the encapsulation composite system. The active ingredients in the fresh fruits and vegetables are at least one of polyphenols, anthocyanins, vitamins, polysaccharides, organic acids, and terpenoids; the natural matrix with encapsulation function is at least one of a natural matrix containing polysaccharides, a natural matrix containing organic acids, and a natural matrix containing terpenoids.

2. The preparation method according to claim 1, characterized in that, The fresh fruits and vegetables mentioned are at least one of the following: blueberries, grapes, mulberries, strawberries, mangoes, pomegranates, broccoli, tomatoes, and pumpkins; The natural matrix with encapsulation function is at least one of the following: Tremella fuciformis, bamboo fungus, Poria cocos, inulin, and sodium alginate. The following amounts of each ingredient are added by weight: 20-40 parts fresh fruits and vegetables, 1-4 parts natural matrix with encapsulation function, and 40-80 parts purified water.

3. The preparation method according to claim 1, characterized in that, In S1, a wet pulverizing device is used for wet pre-pulverization. The rotation speed of the wet pre-pulverization process is 2500~3500 rpm, using 18~22° hammers and toothed rings with 0 cutting depth. The processing time of the wet pre-pulverization process is 10min~20min.

4. The preparation method according to claim 1, characterized in that, In S2, the jet pressure of the high-energy fluid mill is 30-280 MPa.

5. An encapsulated composite system obtained by the preparation method according to any one of claims 1 to 4, characterized in that, The encapsulation complex system includes a natural matrix with encapsulation function and fresh fruits and vegetables. The natural matrix with encapsulation function encapsulates the active ingredients in the fresh fruits and vegetables through intermolecular forces to form a stable complex or microcapsule structure.

6. The embedding composite system according to claim 5, characterized in that, The particle sizes of the encapsulated composite system are as follows: D[3,2] is 13~15 μm, D[4,3] is 34~36 μm, D10 is 5~7 μm, D50 is 26~28 μm, and D90 is 75~78 μm.

7. The application of the encapsulation composite system according to claim 5 in the preparation of functional foods.

8. A functional food and beverage, characterized in that, The functional food and beverage is obtained by sterilization of the encapsulation complex system described in claim 5.

9. The functional food and beverage according to claim 8, characterized in that, The functional food and beverage is a compound blueberry juice, wherein the fresh fruit and vegetable is blueberry, and the natural matrix with encapsulation function is tremella; the compound blueberry juice has an anthocyanin retention rate of no less than 85% after accelerated storage test, and a browning index of no more than 2.5 after 30 days of storage.

10. The application of the preparation method according to any one of claims 1 to 4 in the color protection and stabilization processing of fruit and vegetable juices.

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