A method for preparing liquid bovine colostrum

Abstract

This invention discloses a method for preparing liquid bovine colostrum. The method includes: a defatting step, separating milk raw materials to obtain skim milk and cream; an adsorption chromatography step, treating the skim milk using ion exchange chromatography to obtain chromatographically purified skim milk and eluent components, wherein the eluent used in the ion exchange chromatography has a salt mass fraction greater than 3%; a homogenization step, homogenizing the chromatographically purified skim milk and the cream to obtain a homogenized material; and a mixing step, mixing the homogenized material with the eluent components to obtain the liquid colostrum. This invention reduces the loss of immunoglobulins due to denaturation and inactivation during the processing of bovine colostrum, ensuring their content in bovine colostrum, and ensuring the activity of immunoglobulins in bovine colostrum, further improving the immune properties of the liquid dairy product.

Description

Technical Field

[0001] This invention relates to a method for preparing liquid bovine colostrum, and more specifically to an industrial-scale commercial aseptic liquid whole-milk bovine colostrum preparation method, belonging to the field of dairy product preparation technology. Background Technology

[0002] Currently, liquid bovine colostrum products are mainly produced by defatting bovine colostrum, microfiltration to remove casein, and then enriching immunoglobulins in the permeate (e.g., citing references 1-3), or by simply mixing bovine colostrum with regular milk in a certain ratio (e.g., citing references 4-5), followed by sterilization using ultra-high pressure (a non-large-scale industrial technology) and low-temperature pasteurization. Neither of these two technical solutions can produce industrially viable, commercially available, sterile whole-milk bovine colostrum.

[0003] Conventional ultra-high temperature (UHT) sterilization, typically involving treatment at 135-150°C for 2-5 seconds, is a widely used commercial aseptic technology in the dairy industry. However, this high-temperature treatment can significantly impact the nutritional value of cattle, particularly the heat-sensitive bioactive components in bovine colostrum.

[0004] I. Non-full-dairy cow colostrum suffers severe nutrient loss.

[0005] The overall nutritional value of bovine colostrum is significantly higher than that of its defatted permeate obtained through microfiltration. This difference is not due to a simple summation of components, but rather a systemic advantage determined by the integrity of its multiphase structure, the spatial synergy of its functional components, the enhanced physical stability, and the mechanism of multiplied bioavailability. The following rigorous analysis, based on publicly available literature data, will be presented from multiple dimensions.

[0006] First, at the level of macronutrients and structural functional components, bovine colostrum retains an intact fat globule membrane (MFGM) system, a non-renewable functional platform. Data shows that the MFGM protein content in un-centrifuged bovine colostrum reaches 2.1–3.4 g / L, with core functional proteins such as mucin-1 (MUC1), lactolipoprotein (BTN), and xanthine oxidoreductase (XOR) highly enriched on the lipid bilayer surface. However, the defatting-microfiltration process significantly removes MFGM during centrifugation, resulting in a detection rate of less than 0.05 g / L for MFGM-related proteins in the permeate, exceeding 97%. More importantly, MFGM is not an inert membrane but a natural nanocarrier: its phospholipid bilayer can embed lactoferrin peptides, allowing bound LF to retain 68% antibacterial activity in simulated gastric juice (pH 2.0, 2 h), while the inactivation rate of free LF in the permeate reaches 92% during the same period. This "structural protection-functional enhancement" relationship directly translates into differences in bioavailability. Animal experiments have shown that, when oral administration of the same amount of whole colostrum and permeate, the former has a 2.7-fold higher LF concentration in the ileum tissue (p < 0.01), confirming MFGM's ability to target and deliver active ingredients.

[0007] Secondly, regarding the distribution pattern of immunoglobulins, whole colostrum exhibits a triple spatial anchoring of "micelle phase-whey phase-lipid phase," while the permeate contains only whey phase IgG. The total IgG content in bovine colostrum is 30–80 mg / mL, of which approximately 12–18% (3.6–14.4 mg / mL) is adsorbed onto the surface of casein micelles via electrostatic interactions, and another 5–8% (1.5–6.4 mg / mL) is bound to the phospholipid head group of MFGM. Although 0.1 μm microfiltration allows monomeric IgG (hydrated diameter ~7 nm) to pass through freely, it completely retains both micelle-bound and MFGM-bound IgG. Therefore, the actual IgG concentration in the permeate is only 18–42 mg / mL, a decrease of 35–45% compared to whole milk. More importantly, micelle-bound IgG exhibits stronger stability: after heat treatment at 65 °C for 10 min, the IgG activity retention rate in whole colostrum is 53%, while it is only 28% in the permeate, demonstrating that the micelle structure plays a physical stabilizing role in the IgG conformation.

[0008] Third, regarding the maintenance of the biological activity of growth factors and signaling molecules, epidermal growth factor (EGF) and transforming growth factor-β (TGF-β) in whole colostrum exhibit a clear lipid microenvironment-dependent relationship. Studies have confirmed that EGF partially intercalates into the hydrophobic region of MFGM in whole colostrum, increasing its transmembrane transport efficiency by 3.1 times; TGF-β forms a complex with calcium salts on the surface of casein micelles, prolonging its half-life to 4.2 h (compared to 1.8 h in permeate). Upon removal from the original physical microenvironment, the biological efficacy of growth factors irreversibly declines.

[0009] Finally, in terms of nutritional kinetic parameters, whole colostrum has better osmotic pressure and electrolyte balance, matching the intestinal transport capacity of newborns; while permeate loses calcium salts and causes sodium-potassium imbalance due to defatting and microfiltration.

[0010] Second, conventional commercial aseptic ultra-high temperature (UHT) sterilization processes result in significant losses of heat-sensitive bioactive components.

[0011] Ultra-high temperature (UHT) sterilization, typically referring to treatment at 135-150°C for 2-5 seconds, is a widely used commercial aseptic technology in the dairy industry. However, this high-temperature treatment significantly impacts the nutritional value of bovine milk, particularly heat-sensitive bioactive components in colostrum. The main affected nutrients include immunoglobulins (IgG), lactoferrin, growth factors, and some vitamins and antioxidants. IgG is heat-sensitive and prone to irreversible denaturation and aggregation during UHT treatment. Due to its extremely high temperature, UHT treatment usually leads to a significant reduction in IgG activity, with a loss rate reaching 30-70%. Lactoferrin, a glycoprotein with antibacterial, antiviral, and immunomodulatory functions, is also heat-sensitive. UHT treatment causes the structure of lactoferrin to unfold and become inactive, thereby reducing its bioactivity; under UHT conditions, lactoferrin activity may be completely lost.

[0012] In summary, the nutritional value of bovine colostrum is the result of the synergistic effects of its multiphase structure (lipid phase, micelle phase, and whey phase) and the functional components within it, across spatial, temporal, and physicochemical dimensions. Any separation of a single phase results in irreversible functional decoupling. While the microfiltration permeate enriches some water-soluble active proteins, it sacrifices the four pillars of the MFGM protection system, the micelle stabilization network, and physiological osmotic balance. Therefore, its nutritional value is not a linear subset of whole colostrum, but rather a secondary product with severely incomplete functional dimensions.

[0013] Currently, there is a lack of methods to combine multiple processes to maximize the preservation of the complete nutritional components of bovine colostrum.

[0014] References

[0015] Reference 1 CN120040582A

[0016] Reference 2CN105981908B

[0017] Reference 3CN104434974A

[0018] Reference 4CN101467559A

[0019] Reference 5CN118402555A Summary of the Invention

[0020] The problem the invention aims to solve

[0021] As mentioned earlier, existing technologies can significantly affect the bioactive components of bovine colostrum, reducing its bioactivity. Therefore, there is an urgent need for a combined approach that utilizes multiple processes to overcome the shortcomings of existing technologies that only target single-stage improvements. This approach would form a complete processing system from raw material processing to finished product packaging, minimizing nutrient loss at each stage and thus preserving the full nutritional components of bovine colostrum to the greatest extent possible.

[0022] Solution for solving the problem

[0023] [1]. A method for preparing a liquid colostrum, wherein the method includes:

[0024] The defatting process involves separating the dairy raw materials to obtain skim milk and cream.

[0025] The adsorption chromatography step involves treating the skim milk with ion exchange chromatography to obtain chromatographically purified skim milk and eluent components, wherein the eluent used in the ion exchange chromatography has a salt mass fraction greater than 3%.

[0026] The homogenization step involves homogenizing the skim milk after chromatography with the light cream to obtain a homogenized material.

[0027] The mixing step involves mixing the homogenized material with the elution components to obtain the liquid colostrum;

[0028] Optionally, the colostrum includes at least one of bovine colostrum, sheep colostrum, camel colostrum, and horse colostrum.

[0029] [2]. According to the method described in [1], in the step of the adsorption chromatography treatment, the skim milk is subjected to cation exchange chromatography treatment to obtain a flow-through liquid, and eluted with an elution liquid to obtain the cation adsorption eluent component, wherein the linear flow rate of the skim milk in the cation exchange chromatography treatment is less than 1200 cm / h.

[0030] [3]. The method according to [1] or [2], wherein the cationic adsorption eluent comprises basic protein;

[0031] Optionally, the basic protein includes at least one of lactoferrin, β-lactoglobulin, bovine serum albumin, and peroxidase.

[0032] [4]. According to the method of [2] or [3], wherein, in the step of the adsorption chromatography treatment, the flow-through liquid obtained by the cation exchange chromatography treatment is subjected to anion exchange chromatography treatment to obtain the chromatographically defatted emulsion, and the anion adsorption eluent is eluted with an elution buffer to obtain the anion adsorption eluent component, wherein the linear flow rate of the flow-through liquid in the anion exchange chromatography treatment is less than 300 cm / h.

[0033] [5]. The method according to any one of [1]-[4], wherein the anion adsorption eluent includes nutrients;

[0034] Optionally, the nutrients include immunoglobulins and / or bovine serum albumin.

[0035] [6]. The method according to any one of [1]-[5], wherein, in the step of the adsorption chromatography treatment, the temperature of the skim milk is below 25°C;

[0036] Preferably, the temperature of the skim milk is below 18°C.

[0037] [7]. The method according to any one of [1]-[6], wherein, prior to the homogenization step, the method further includes a step of ultrafiltration of the elution components;

[0038] Preferably, the ultrafiltration process includes solution displacement and / or concentration of the eluted components.

[0039] [8]. The method according to any one of [1]-[7], wherein the method further includes sterilization treatment, wherein the sterilization treatment includes sterilizing skim milk, ultrafiltration material and / or homogenized material.

[0040] [9]. A liquid promulgation prepared according to any one of [1]-[8].

[0041]

[10] . A formulated dairy product, wherein the formulated dairy product comprises: a liquid colostrum prepared according to any one of the preparation methods described in [1]-[8] or a liquid colostrum as described in [9],

[0042] Optionally, the formulated dairy product may also contain one or more of the following additional components:

[0043] (A) Functional protein components, (B) Carbohydrate components, (C) Fats and oils, (D) Mineral salts, (E) Other nutritional supplement components.

[0044] The effects of the invention

[0045] This invention reduces the loss of immunoglobulins due to denaturation and inactivation during the processing of bovine colostrum, ensuring their content in bovine colostrum and maintaining their activity in bovine colostrum, thereby further improving the immune properties of liquid dairy products.

[0046] This invention proposes an industrial-scale commercial aseptic liquid bovine colostrum preparation method by employing processes such as centrifugation, anion and cation chromatography, microfiltration membrane sterilization, high-temperature instantaneous immersion sterilization, and aseptic liquid preparation, which maximizes the preservation of all nutritional components of bovine colostrum.

[0047] The method provided by this invention preserves the complete nutritional components of bovine colostrum to the greatest extent possible. The whole colostrum exhibits stable triple spatial anchoring in the "micelle phase-whey phase-lipid phase", retains a more complete fat globule membrane (MFGM) system, maintains better biological activity of growth factors and signaling molecules, and has a higher content of trace elements. Attached Figure Description

[0048] Figure 1 This is the technical approach of the present invention.

[0049] Figures 2A-2D The chromatograms are for the examples and comparative examples.

[0050] Figures 3A-3D The images shown are polyacrylamide gel electrophoresis diagrams for the examples and comparative examples. Detailed Implementation

[0051] Various exemplary embodiments, features, and aspects of the present invention will be described in detail below. The term "exemplary" as used herein means "serving as an example, embodiment, or illustration." Any embodiment described herein as "exemplary" is not necessarily to be construed as superior to or better than other embodiments.

[0052] Furthermore, to better illustrate the present invention, numerous specific details are set forth in the following detailed embodiments. Those skilled in the art should understand that the present invention can be practiced without certain specific details. In other instances, methods, means, apparatus, and steps well known to those skilled in the art have not been described in detail in order to highlight the spirit of the present invention.

[0053] Unless otherwise stated, all units used in this specification are international standard units, and all numerical values ​​and ranges appearing in this invention should be understood to include systematic errors that are unavoidable in industrial production.

[0054] In this specification, the word "may" has two meanings: to perform a certain process and not to perform a certain process.

[0055] In this specification, references to "some specific / preferred embodiments," "other specific / preferred embodiments," "implementation," etc., refer to specific elements (e.g., features, structures, properties, and / or characteristics) related to that embodiment, which are included in at least one of the embodiments described herein and may or may not be present in other embodiments. Furthermore, it should be understood that these elements may be combined in any suitable manner in various embodiments.

[0056] In this specification, the range of values ​​referred to as "value A to value B" refers to the range including the endpoint values ​​A and B.

[0057] In this invention, the term "about" is used to define that the numerical ranges and parameters of this invention are approximate values, while specific related values ​​have been presented as precisely as possible. Unless otherwise explicitly stated, it should be understood that all ranges, quantities, values, and percentages used in this invention are modified by "about". Here, "about" generally means that the actual value is within ±3%, ±2%, ±1%, or ±0.5% of a specific value or range.

[0058] In this invention, the term "infant" is used to refer to the human group aged 0 to 6 months.

[0059] In this invention, the term "older infant" refers to the human group aged 6 to 12 months.

[0060] In this invention, the term "infant" is used to refer to the human group aged 12 to 36 months.

[0061] In this invention, the term "infant" refers to the human group under the age of 3 years.

[0062] In this invention, the term "children" refers to a group of human beings who are older than 3 years and younger than 12 years and are in the growth and development stage.

[0063] In this manual, the term "adult" refers to a person who is 18 years of age or older.

[0064] In this manual, the term "teenager" refers to people aged 7-40.

[0065] In this manual, the term "middle-aged person" refers to a person aged 41-65.

[0066] In this manual, the term "elderly person" or "senior citizen" refers to a person aged 65 or older. In this manual, the numerical range indicated by "above" or "below" refers to a range that includes the stated number.

[0067] In this specification, the terms "optional" or "optional" are used to indicate the use or omission of certain substances, components, procedures, application conditions, etc.

[0068] All unit names used in this manual are international standard unit names.

[0069] <First Aspect>

[0070] A first aspect of the present invention provides a method for preparing a liquid colostrum, the method comprising:

[0071] The defatting process involves separating the dairy raw materials to obtain skim milk and cream.

[0072] The adsorption chromatography step involves treating the skim milk with ion exchange chromatography to obtain chromatographically purified skim milk and eluent components, wherein the eluent used in the ion exchange chromatography has a salt mass fraction greater than 3%.

[0073] The homogenization step involves homogenizing the skim milk after chromatography with the light cream to obtain a homogenized material.

[0074] The mixing step involves mixing the homogenized material with the elution components to obtain the liquid colostrum.

[0075] In some implementations, the colostrum includes at least one of bovine colostrum, sheep colostrum, camel colostrum, and horse colostrum.

[0076] In some exemplary embodiments, the colostrum includes bovine colostrum.

[0077] (Dairy raw materials)

[0078] In this specification, the term "milk" is used to refer to the fluid obtained from the mammary glands of a mammal in the process of lactation. The term "milk" should be interpreted broadly and encompasses both raw milk (i.e., the fluid obtained directly from the mammary glands) and standardized dairy products (such as skim milk or whole milk).

[0079] In some embodiments, the milk raw materials described in this invention include milk raw materials derived from animal sources, typically including milk derived from cows, sheep, camels, deer, or horses.

[0080] In some preferred embodiments, the milk raw material of the present invention is milk derived from cattle.

[0081] In some exemplary embodiments, the milk raw material includes thawing and dissolving solid bovine colostrum into a liquid, and then pumping it out and cooling it before storing it in a storage tank at a temperature of 0-7°C after the temperature reaches 10-15°C.

[0082] In some preferred embodiments, fresh liquid bovine colostrum can be used directly.

[0083] (Defatting treatment)

[0084] In some embodiments, the defatting process includes separating the dairy raw materials to obtain skim milk and cream.

[0085] In some specific implementation schemes, from the perspective of convenience in subsequent preparation, in addition to using raw milk, the above-mentioned milk raw materials can preferably be processed through processes such as concentration and defatting to obtain milk with reduced fat content.

[0086] In some implementations, the skim milk has a fat content of less than 0.2%.

[0087] In some preferred embodiments, the fat content of the skim milk is between 0.05% and 0.1%, for example: 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, etc.

[0088] In some specific implementation schemes, the bovine colostrum in the storage tank is pumped into the heat recovery section of a three-stage plate heat exchanger to preheat the degreased and sterilized product, and then enters the heating section to be heated to 50℃-55℃ before being degreased by a centrifuge, reducing the fat content to 0.05-0.1%.

[0089] In some implementations, skim milk also undergoes degreasing and sterilization treatment.

[0090] In some specific implementation schemes, skim milk undergoes microfiltration for oil removal and sterilization.

[0091] In some implementations, skim milk is separated by a microfiltration membrane with a pore size of less than 5 μm, preferably less than 2 μm, to remove oil and bacteria.

[0092] Those skilled in the art can select microfiltration membranes of materials such as organic membranes (polypropylene (PP), polyvinylidene fluoride (PVDF), polyethersulfone (PES), etc.) and inorganic membranes (ceramic membranes and metal membranes), and separate skim milk through the microfiltration membrane under appropriate driving pressure to remove oil and bacteria.

[0093] In some specific implementation schemes, the centrifuged and defatted bovine colostrum is separated by a 1.4μm pore size microfiltration membrane. The degreased and sterilized material is returned to the three-stage plate heat recovery section of the previous centrifugation and defatting process to preheat the feed material. Then, the milk is cooled to 10-15°C in the cooling section and stored in a buffer tank for later use.

[0094] (Adsorption chromatography treatment)

[0095] In some embodiments, the adsorption chromatography step involves treating the skim milk with ion exchange chromatography to obtain chromatographically purified skim milk and eluent components, wherein the mass fraction of salt in the eluent is greater than 3% in the ion exchange chromatography step.

[0096] In some embodiments, in the adsorption chromatography step, the skim milk is subjected to cation exchange chromatography to obtain a flow-through liquid, and eluted with an eluent to obtain the cationic adsorbed eluent components, wherein the linear flow rate of the skim milk in the cation exchange chromatography is less than 1000 cm / h.

[0097] In some embodiments, the eluent is a salt solution. Examples include buffer solutions or buffered salt solutions such as sodium chloride solution, potassium chloride solution, ammonium sulfate solution, sodium sulfate solution, and phosphate buffer solution.

[0098] In some specific embodiments, the eluent is a NaCl solution.

[0099] In some preferred embodiments, the salt mass fraction in the eluent is greater than 4%.

[0100] In some preferred embodiments, the mass fraction of salt in the eluent is less than 7%.

[0101] In some specific implementations, the mass fraction of salt in the eluent is around 5.0-6.0%, such as 5.0%, 5.5%, 6.0%, etc.

[0102] In some embodiments, the skim milk is treated at a temperature below 25°C during the cation exchange chromatography step.

[0103] In some preferred embodiments, the skim milk is processed at a temperature below 18°C.

[0104] In some preferred embodiments, the skim milk treatment temperature is between 10-15°C, for example: 10°C, 11°C, 12°C, 13°C, 14°C, 15°C, etc.

[0105] In some preferred embodiments, the linear flow rate of the skim milk in the cation exchange chromatography process is 500-900 cm / h, for example: 500 cm / h, 600 cm / h, 700 cm / h, 800 cm / h, 900 cm / h, etc.

[0106] In some preferred embodiments, the column height of the skim milk in the cation exchange chromatography process is between 18 and 28 cm, for example: 18 cm, 20 cm, 22 cm, 25 cm, 26 cm, 28 cm, etc.

[0107] In some embodiments, the cation adsorption eluent includes a basic protein.

[0108] In some optional embodiments, the basic protein includes at least one of lactoferrin, β-lactoglobulin, bovine serum albumin, and peroxidase.

[0109] In this invention, as long as it is possible to obtain cation adsorption and elution components including basic proteins, there are no special restrictions on the specific type of cation exchange resin, etc. Those skilled in the art can choose a suitable cation exchange resin for cation exchange chromatography.

[0110] In some specific implementations, the cation adsorption involves feeding skim milk into a primary chromatography column via a degassing tank for adsorption chromatography with the cation exchange resin inside the column. The skim milk temperature is 10-15℃, the linear flow rate is 500-900 cm / h, and the column height is 18-28 cm. After chromatography, the column is first flushed with RO water to remove the skim milk from the resin before being drained. Then, a NaCl solution with a mass fraction of approximately 5.0-6.0% is used to elute the resin-affinity basic proteins such as lactoferrin and peroxidase.

[0111] In some embodiments, in the adsorption chromatography step, the flow-through obtained by cation exchange chromatography is subjected to anion exchange chromatography to obtain the chromatographically defatted emulsion, and the anion adsorbed eluent is eluted with an elution buffer to obtain the anion adsorbed eluent components, wherein the linear flow rate of the flow-through in the anion exchange chromatography is less than 300 cm / h.

[0112] In some embodiments, the eluent is a salt solution. Examples include buffer solutions or buffered salt solutions such as sodium chloride solution, potassium chloride solution, ammonium sulfate solution, sodium sulfate solution, and phosphate buffer solution.

[0113] In some specific embodiments, the eluent is a NaCl solution.

[0114] In some preferred embodiments, the salt mass fraction in the eluent is greater than 4%.

[0115] In some preferred embodiments, the mass fraction of salt in the eluent is less than 7%.

[0116] In some specific implementations, the mass fraction of salt in the eluent is around 5.0-6.0%, such as 5.0%, 5.5%, 6.0%, etc.

[0117] In some embodiments, the skim milk is treated at a temperature below 25°C during the anion exchange chromatography step.

[0118] In some preferred embodiments, the skim milk is processed at a temperature below 18°C.

[0119] In some preferred embodiments, the skim milk treatment temperature is between 10-15°C, for example: 10°C, 11°C, 12°C, 13°C, 14°C, 15°C, etc.

[0120] In some preferred embodiments, the linear flow rate of the flow-through liquid in the anion exchange chromatography process is 50-120 cm / h, for example: 50 cm / h, 70 cm / h, 80 cm / h, 90 cm / h, 100 cm / h, 120 cm / h, etc.

[0121] In some preferred embodiments, the column height of the flow-through liquid in the anion exchange chromatography process is between 18 and 28 cm, for example: 18 cm, 20 cm, 22 cm, 25 cm, 26 cm, 28 cm, etc.

[0122] In some embodiments, the anion adsorption elution component includes nutrients.

[0123] In some optional embodiments, the nutrients include immunoglobulins and / or bovine serum albumin.

[0124] In this invention, as long as it is possible to obtain anion adsorption and elution components including nutrients, there are no special restrictions on the specific type of anion exchange resin, etc. Those skilled in the art can choose a suitable anion exchange resin for anion exchange chromatography.

[0125] In some specific implementations, the anion adsorption includes, after primary chromatography, the defatted emulsion is degassed and fed into a secondary chromatography column to undergo adsorption chromatography with the anion exchange resin inside the column. The defatted emulsion temperature is 10-15℃, the linear flow rate is 50-120cm / h, and the column height is 18-28cm. After chromatography, the column is first flushed with RO water to remove the defatted emulsion from the resin and then drained. Then, the resin-affinity immunoglobulins, bovine serum albumin, etc., are eluted with a NaCl solution with a mass fraction of about 5.0-6.0%.

[0126] (Ultrafiltration and sterilization treatment)

[0127] In some embodiments, the elution components are further subjected to ultrafiltration before the homogenization step.

[0128] In some preferred embodiments, the ultrafiltration process includes solution displacement and / or concentration of the eluted components.

[0129] In some preferred embodiments, the solution replacement includes desalting the elution component.

[0130] The present invention uses ultrafiltration to improve the purity of the eluted protein components and protect the activity of nutrients.

[0131] In some preferred embodiments, the molecular weight cutoff of the ultrafiltration treatment is between 10-50 kD, for example: 10 kD, 20 kD, 30 kD, 40 kD, 50 kD, etc.

[0132] In some specific implementations, the ultrafiltration process includes eluting proteins with a NaCl solution of about 5.0-6.0% by mass and then pumping them through a pipeline into a 10-50kD ultrafiltration system for desalting and concentration, followed by temporary storage in an ultrafiltration tank.

[0133] In some embodiments, the ultrafiltration process further includes sterilization of the ultrafiltration material.

[0134] In some preferred embodiments, the sterilization process includes membrane sterilization.

[0135] In some specific implementation schemes, the ultrafiltration temporary storage material is sterilized by a sterilization membrane with a pore size of 0.6 ± 0.2 μm, and then injected into the sterile solution preparation system for later use.

[0136] (Homogeneous)

[0137] In some implementations, the homogenization step involves homogenizing the skim milk after chromatography with the cream to obtain a homogenized material.

[0138] In some embodiments, during the homogenization step, the skim milk after chromatography and the cream are heated to below 60°C.

[0139] In some preferred embodiments, during the homogenization step, the skim milk after chromatography and the cream are heated to 48-55°C, for example: 48°C, 50°C, 52°C, 53°C, 54°C, 55°C, etc.

[0140] In some preferred embodiments, the homogenization pressure during the homogenization step is 120.0-240.0 bar, for example: 120.0 bar, 150.0 bar, 200.0 bar, 220.0 bar, 230.0 bar, 240.0 bar, etc.

[0141] In some specific implementations, the homogenization step includes starting a homogenization program, mixing skim milk and cream after chromatography, heating to 48-55°C, and then entering a homogenizer. The homogenization process is carried out at a pressure of 120.0-240.0 bar.

[0142] (Sterilization treatment)

[0143] In some embodiments, the method further includes a sterilization process, wherein the sterilization process includes sterilizing skim milk, ultrafiltration material, and / or homogenized material.

[0144] In some specific implementations, the homogenized material is sterilized, and the sterilization includes ultra-high temperature instantaneous invasive sterilization.

[0145] In some preferred embodiments, the ultra-high temperature instantaneous invasive sterilization conditions include a sterilization temperature of 127°C-135°C for a duration of 1-3 seconds, for example: a sterilization temperature of 135°C for a duration of 1 second, or a sterilization temperature of 127°C for a duration of 3 seconds.

[0146] In some specific implementations, the ultra-high temperature instantaneous invasive sterilization includes injecting homogenized bovine colostrum into an ultra-high temperature instantaneous invasive sterilization system, with sterilization conditions of 127-135°C for 1-3 seconds, followed by cooling to 10-15°C and injecting it into an aseptic liquid preparation system to mix with the aforementioned ultrafiltration-treated material.

[0147] (mix)

[0148] In some embodiments, the mixing step involves mixing the homogenized material with the eluted components to obtain liquid bovine colostrum.

[0149] In some exemplary embodiments, the mixing step involves mixing the sterilized homogenized material with the sterilized ultrafiltration material to obtain liquid bovine colostrum.

[0150] In some implementations, the mixing step takes no more than 10 minutes.

[0151] In some specific implementations, the mixing step includes thoroughly mixing all materials in a sterile dispensing system for a mixing time of not less than 3 minutes.

[0152] The following are exemplary embodiments of the present invention:

[0153] 1. Bovine colostrum acceptance: After the frozen bovine colostrum blocks arrive at the factory, samples are taken for testing according to the quality control plan. After arrival, the colostrum blocks are stored in a cold storage at a temperature of ≤-14.5℃. The released bovine colostrum blocks are crushed in a crusher and then enter a dissolving tank. The solid bovine colostrum is thawed and dissolved into liquid by heating with hot water on the tank wall and stirring. After the temperature reaches 10-15℃, it is pumped out and cooled by a plate heat exchanger before being sent to a storage tank for storage. The storage temperature of bovine colostrum is 0-7℃.

[0154] 2. Centrifugal defatting: The bovine colostrum stored in the storage tank in step 1 is pumped into the heat recovery section of a three-stage plate heat exchanger. After being preheated by the microfiltration, oil removal, sterilization, and defatting processes in step 3, the product then enters the heating section and is heated to 50℃-55℃ before being defatted by a centrifuge (centrifuge speed 3500-4500r / min). The fat content is reduced to 0.05-0.1%, yielding centrifuged defatted bovine colostrum (skimmed milk) and light cream.

[0155] 3. Microfiltration for oil removal and sterilization: The bovine colostrum obtained in step 2 after centrifugation and defatting is separated by a 1.4μm pore size microfiltration membrane. The oil-removed and sterilized material is returned to the three-stage plate heat recovery section of the centrifugation and defatting process in step 2 to preheat the feed material in step 2. Then, the defatted milk that has undergone microfiltration for oil removal and sterilization is cooled to 10-15℃ in the cooling section and stored in a buffer tank for later use.

[0156] 4. Cation chromatography: The skim milk that has been degreased and sterilized by microfiltration in the buffer tank of step 3 is pumped into a primary chromatography column via a degassing tank for adsorption chromatography with the cation exchange resin in the column (particle size 150-400um). The temperature of the skim milk is 10-15℃, the linear flow rate is 500-900cm / h, and the column height is 18-28cm. After chromatography, the flow-through liquid (i.e., the skim milk after primary chromatography) is collected. Then, it is first flushed with RO water to remove the skim milk from the resin and discharged to the ground. Then, the resin-affinity alkaline proteins such as lactoferrin and peroxidase (cation adsorption eluent components) are eluted with a NaCl solution with a mass fraction of about 5.0-6.0%. The eluent is then pumped into a 10-50kD ultrafiltration system for desalting and concentration. After concentration, it is temporarily stored in an ultrafiltration temporary storage tank.

[0157] 5. Anion chromatography: The skimmed milk obtained from the first chromatography step 4 is degassed and fed into a second chromatography column to undergo adsorption chromatography with the anion exchange resin (particle size 150-400 μm). The skimmed milk temperature is 10-15℃, the linear flow rate of the skimmed milk after the first chromatography is 50-120 cm / h, and the column height is 18-28 cm. After chromatography, the flow-through liquid (i.e., the skimmed milk from the second chromatography or the skimmed milk after chromatography) is collected. Then, it is first flushed with RO water to remove the skimmed milk from the resin and discharged to the ground. Then, the resin-affinity immunoglobulins, bovine serum albumin, etc. (anion adsorption eluent components) are eluted with a NaCl solution with a mass fraction of about 5.0-6.0%. The eluent is then fed into a 10-50 kD ultrafiltration system for desalting and concentration. After concentration, it is temporarily stored in an ultrafiltration storage tank.

[0158] 6. Microfiltration sterilization: The materials temporarily stored in the ultrafiltration temporary storage tank in step 5 (elution components, i.e., cation adsorption eluent components and anion adsorption eluent components) are sterilized through a sterilization membrane with a pore size of 0.6 + 0.2 μm. After sterilization, they are injected into the aseptic solution preparation system for later use.

[0159] 7. Homogenization: Start the homogenization program, combine the skim milk after chromatography in step 5 (secondary chromatography skim milk) with the light cream obtained after centrifugation and skimming in step 2, heat to 48-55℃, and put into the homogenizer. The homogenization pressure during the homogenization process is 120.0-240.0 bar to obtain homogenized bovine colostrum.

[0160] 8. Ultra-high temperature instantaneous invasive sterilization: The homogenized bovine colostrum obtained in step 7 is injected into an ultra-high temperature instantaneous invasive sterilization system. The sterilization conditions are 127-135℃ for 1-3 seconds. After sterilization, the temperature is reduced to 10-15℃ and injected into an aseptic liquid preparation system to mix with the sterilized material in step 6.

[0161] 9. Aseptic solution preparation: All materials are thoroughly mixed in the aseptic solution preparation system for a mixing time of ≥3 minutes;

[0162] 10. Aseptic filling: The uniformly mixed materials are fed into the aseptic filling system for commercial aseptic filling;

[0163] 11. Coding and Packing: Coding, packing, and finally warehousing.

[0164] <Second aspect>

[0165] The second aspect of the present invention provides a liquid primary emulsion prepared by the preparation method described in the first aspect of the present invention.

[0166] In some exemplary embodiments, the liquid colostrum is bovine colostrum.

[0167] <Third aspect>

[0168] A third aspect of the present invention provides a formulated dairy product, the formulated dairy product comprising liquid colostrum prepared by the preparation method described in the first aspect of the present invention or liquid colostrum described in the second aspect of the present invention.

[0169] The dairy products formulated according to the present invention can be in solid, liquid or semi-solid form, preferably in liquid form.

[0170] In some optional embodiments, the formulated dairy product may also contain one or more of the following additional components:

[0171] (A) Functional protein components, (B) Carbohydrate components, (C) Fats and oils, (D) Mineral salts, (E) Other nutritional supplement components.

[0172] (Functional protein components)

[0173] There are generally no special restrictions on the functional protein ingredients that can be used; for example, various whey proteins, immunoglobulins, lactoferrin, etc. There are also no special restrictions on the amount of these ingredients added, as long as they comply with laws and regulations.

[0174] (Carbohydrate composition)

[0175] There are no particular restrictions on the type or source of carbohydrates added to the formula dairy products of this invention; those carbohydrates commonly used in formula dairy products in this field can be used.

[0176] In some specific embodiments, the carbohydrate component described in this invention mainly refers to sugars. These sugars are typically a general term for polyhydroxy aldehydes or polyhydroxy ketones and their condensation polymers and certain derivatives, generally composed of carbon, hydrogen, and oxygen. All sugars can be written with the empirical molecular formula: Cn(H₂O). n .

[0177] In this invention, the carbohydrates typically include monosaccharides, disaccharides, polysaccharides, or oligosaccharides.

[0178] Monosaccharides can mainly include glucose, fructose, etc.

[0179] Disaccharides, polysaccharides, or oligosaccharides can include: sucrose, lactose, fructotriose, fructotetraose, fructopentose, fructooligosaccharides, glucosamine, (maltodextrin), and various forms of human milk oligosaccharides.

[0180] In some specific implementations, the carbohydrates may be added in the form of dietary fiber. Examples of dietary fiber substances include one or more of the following: inulin, konjac flour, galactooligosaccharides, fructooligosaccharides, isomaltooligosaccharides, soybean polysaccharides, cyclodextrin, maltodextrin, resistant dextrin, and soybean fiber.

[0181] Furthermore, there is no particular limitation on the total carbohydrate content in the formula dairy products of the present invention, and it can be formulated with reference to the carbohydrate content commonly used in infant formula milk powder products for the middle-aged and elderly in the art.

[0182] (Oil and fat components)

[0183] There are no particular restrictions on the types of oils added to the formula dairy products of this invention. Fatty acid glycerides commonly used in infant formula and formula milk powder products for the middle-aged and elderly can be used.

[0184] These glycerides can be obtained from plant extraction or through artificial synthesis (transesterification). Transesterification can be achieved by esterification of glycerol with fatty acids in the presence of a catalyst, or by transesterification of triglycerides and fatty acids from various existing sources in the presence of a (specific) catalyst (enzyme). For plant oils extracted from plants, the desired fatty acid glycerides can be obtained by mixing one or more of the following plant oils: rapeseed oil, soybean oil, sunflower oil, olive oil, sesame oil, corn oil, flaxseed oil, and camellia seed oil.

[0185] The fatty acid glycerides described in this invention are primarily triglycerides. Examples of such glycerides include OPL, OPO, MLCT, OOL, OPP, OLO, OLL, LPL, LPLn, OPLn, LPCa, and OPCa.

[0186] The term "MLCT structured mixed ester" refers to a structurally mixed ester whose main components are medium- to long-chain fatty acid triglycerides. "MLCT" stands for Middle to Long Chain Triglycerides, a structural ester whose glycerol backbone contains both medium-chain fatty acids (M) and long-chain fatty acids (L). Medium-chain fatty acids are those containing 6 to 12 carbon atoms, while long-chain fatty acids are those with more than 12 carbon atoms. MLCTs possess the characteristics of both medium- and long-chain fatty acids, offering the advantages of supplementing essential fatty acids, providing rapid energy, and not causing fat accumulation. Common MLCTs include OPLa, LPLa, OPCa, OLaO, OLaL, OMLa, SLaL, and SLaO.

[0187] (Mineral salt components)

[0188] The mineral salts that can be added to the dairy products formulated in this invention are mainly used to introduce micronutrients, including iron, copper, manganese, zinc, cobalt, molybdenum, chromium, nickel, vanadium, fluorine, selenium, iodine, silicon, tin, etc.

[0189] Furthermore, regarding the content of mineral salts in formula dairy products, as long as it complies with the provisions of laws and regulations, it is acceptable.

[0190] (Other nutritional supplement ingredients)

[0191] There are no particular restrictions on other nutritional supplement ingredients that can be added to the dairy products formulated according to the present invention, and they can be prepared in accordance with existing methods in the art.

[0192] In some specific implementations, these supplement ingredients include one or more selected from vitamins, probiotics, and unsaturated fatty acids.

[0193] Examples of vitamins include one or more of the following: vitamin A, beta-carotene, vitamin D3, vitamin E, vitamin K1, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, pantothenic acid, folic acid, niacin, and biotin.

[0194] For probiotics, for example, there are Bifidobacteria probiotics that are beneficial to the gut.

[0195] Examples of unsaturated fatty acids include arachidonic acid and docosahexaenoic acid.

[0196] Furthermore, there are no particular restrictions on the source of each of the components (A) to (E) above; for example, they can be introduced by mixing with milk or other nutritional additives.

[0197] Examples, Comparative Examples and Test Cases

[0198] The embodiments of the present invention will be described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of the invention. Unless otherwise specified in the examples, conventional conditions or conditions recommended by the manufacturer are followed. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0199] Example 1

[0200] 1. Acceptance of bovine colostrum: After the frozen bovine colostrum blocks arrive at the factory, samples are taken for testing according to the quality control plan. After arrival, the colostrum blocks are stored in a cold storage at a temperature of -18℃. The released bovine colostrum blocks are crushed in a crusher and then put into a dissolving tank. The solid bovine colostrum is thawed and dissolved into liquid by heating with hot water on the tank wall and stirring. After the temperature reaches 10℃, it is pumped out and cooled by a plate heat exchanger before being sent to a storage tank for storage. The storage temperature of bovine colostrum is 4℃.

[0201] 2. Centrifugal defatting: The bovine colostrum stored in the storage tank in step 1 is pumped into the heat recovery section of a three-stage plate heat exchanger. After being preheated by the product that has been micro-filtered for oil removal and sterilization in step 3, it enters the heating section and is heated to 50°C. Then, it is defatted by a centrifuge (centrifuge speed 4500r / min) until the fat content is reduced to 0.05-0.1%, obtaining centrifuged defatted bovine colostrum (skimmed milk) and light cream.

[0202] 3. Microfiltration for oil removal and sterilization: The centrifuged and defatted bovine colostrum obtained in step 2 is separated by a 1.4μm pore size microfiltration membrane (Tongzhou Zongheng (Xiamen) Fluid Technology Co., Ltd., polyethersulfone PES membrane). The degreased and sterilized material is returned to the three-stage plate heat recovery section of the centrifugal defatting in step 2 to preheat the feed material in step 2. Then, the defatted milk that has been degreased and sterilized by microfiltration is cooled to 10°C in the cooling section and stored in the buffer tank for later use.

[0203] 4. Cation chromatography: The skim milk that has been degreased and sterilized by microfiltration in the buffer tank of step 3 is pumped into a primary chromatography column via a degassing tank and adsorbed onto the cation exchange resin (Chutian Microsphere Biotechnology (Changsha) Co., Ltd., particle size 150um) in the column for adsorption chromatography. The temperature of the skim milk is 10℃, the linear flow rate of the skim milk is 500cm / h, and the column height is 18cm. After the chromatography is completed, the flow-through liquid (i.e., the skim milk after the primary chromatography) is collected. Then, it is first flushed with RO water to remove the skim milk from the resin and discharged to the ground. Then, the resin-affinity alkaline proteins such as lactoferrin and peroxidase (cation adsorption eluent components) are eluted with a NaCl solution with a mass fraction of about 5.0%. The eluent is then pumped into a 10kD ultrafiltration system for desalting and concentration. After concentration, it is temporarily stored in an ultrafiltration temporary storage tank.

[0204] 5. Anion chromatography: The skimmed emulsion obtained in step 4 after primary chromatography is pumped into a secondary chromatography column via a degassing tank and subjected to adsorption chromatography with the anion exchange resin in the column (Chutian Microsphere Biotechnology (Changsha) Co., Ltd., particle size 150um). The temperature of the skimmed emulsion is 10℃, the linear flow rate of the skimmed emulsion after primary chromatography is 50cm / h, and the column height is 18cm. After chromatography, the flow-through liquid (i.e., the skimmed emulsion after secondary chromatography, or the skimmed emulsion after chromatography) is collected. Then, it is first flushed with RO water to remove the skimmed emulsion from the resin and discharged to the ground. Then, the resin-affinity immunoglobulins, bovine serum albumin, etc. (anion adsorption eluent components) are eluted with a NaCl solution with a mass fraction of about 5.0%. The eluent is then pumped into a 10kD ultrafiltration system for desalting and concentration. After concentration, it is temporarily stored in an ultrafiltration temporary storage tank.

[0205] 6. Microfiltration sterilization: The material temporarily stored in the ultrafiltration storage tank in step 5 (elution components, i.e., cation adsorption eluent components and anion adsorption eluent components) is sterilized through a 0.6+0.2μm pore size double-layer sterilization membrane (integrated composite membrane, including a 0.6μm membrane and a 0.2μm membrane, made of polyethersulfone, the same below), and then injected into the aseptic solution preparation system for later use;

[0206] 7. Homogenization: Start the homogenization program, combine the skim milk after step 5 chromatography (secondary chromatography skim milk) with the light cream obtained after centrifugation and skimming in step 2, heat to 48°C, and put into the homogenizer. The homogenization pressure is 120.0 bar to obtain homogenized bovine colostrum.

[0207] 8. Ultra-high temperature instantaneous invasive sterilization: The homogenized bovine colostrum obtained in step 7 is injected into an ultra-high temperature instantaneous invasive sterilization system. The sterilization conditions are 127°C for 3 seconds. After sterilization, the temperature is reduced to 10°C and injected into an aseptic liquid preparation system to mix with the sterilized material in step 6.

[0208] 9. Aseptic solution preparation: All materials are thoroughly mixed in the aseptic solution preparation system for 3 minutes;

[0209] 10. Aseptic filling: The uniformly mixed materials are fed into the aseptic filling system for commercial aseptic filling;

[0210] 11. Coding and Packing: Coding, packing, and finally warehousing.

[0211] Example 2:

[0212] 1. Acceptance of bovine colostrum: After the frozen bovine colostrum blocks arrive at the factory, samples are taken for testing according to the quality control plan. After arrival, the colostrum blocks are stored in a cold storage at a temperature of -18℃. The released bovine colostrum blocks are crushed in a crusher and then put into a dissolving tank. The solid bovine colostrum is thawed and dissolved into liquid by heating with hot water on the tank wall and stirring. After the temperature reaches 15℃, it is pumped out and cooled by a plate heat exchanger before being sent to a storage tank for storage. The storage temperature of bovine colostrum is 4℃.

[0213] 2. Centrifugal defatting: The bovine colostrum stored in the storage tank in step 1 is pumped into the heat recovery section of a three-stage plate heat exchanger. After being preheated by the product that has undergone microfiltration and sterilization in step 3, it enters the heating section and is heated to 55°C. Then, it is defatted by a centrifuge (centrifuge speed 3500r / min) until the fat content is reduced to 0.05-0.1%, obtaining centrifuged defatted bovine colostrum (skimmed milk) and light cream.

[0214] 3. Microfiltration for oil removal and sterilization: The bovine colostrum obtained in step 2 after centrifugation and defatting is separated by a 1.4μm pore size microfiltration membrane. The oil-removed and sterilized material is returned to the three-stage plate heat recovery section of the centrifugation and defatting process in step 2 to preheat the feed material in step 2. Then, the defatted milk that has been microfiltration for oil removal and sterilization is cooled to 15°C in the cooling section and stored in a buffer tank for later use.

[0215] 4. Cation chromatography: The skim milk that has been microfiltered and sterilized in the buffer tank of step 3 is pumped into a primary chromatography column via a degassing tank for adsorption chromatography with the cation exchange resin in the column (particle size 400 μm). The temperature of the skim milk is 15℃. The linear flow rate of the skim milk that has been microfiltered and sterilized in step 3 is 900 cm / h, and the column height is 28 cm. After chromatography, the flow-through liquid (i.e., the skim milk after primary chromatography) is collected. Then, it is first flushed with RO water to remove the skim milk from the resin and discharged to the ground. Then, the resin-affinity alkaline proteins such as lactoferrin and peroxidase (cation adsorption eluent components) are eluted with a NaCl solution with a mass fraction of about 6.0%. The eluent is then pumped into a 50 kD ultrafiltration system for desalting and concentration. After concentration, it is temporarily stored in an ultrafiltration temporary storage tank.

[0216] 5. Anion chromatography: The skim milk obtained from the first chromatography in step 4 is pumped into a second chromatography column via a degassing tank for adsorption chromatography with the anion exchange resin in the column (particle size 400 μm). The skim milk temperature is 15℃, the linear flow rate of the skim milk after the first chromatography is 120 cm / h, and the column height is 28 cm. After chromatography, the flow-through liquid (i.e., the skim milk from the second chromatography or the skim milk after chromatography) is collected. Then, it is first flushed with RO water to remove the skim milk from the resin and discharged to the ground. Then, the resin-affinity immunoglobulins, bovine serum albumin, etc. (anion adsorption eluent components) are eluted with a NaCl solution with a mass fraction of about 6.0%. The eluent is then pumped into a 50 kD ultrafiltration system for desalting and concentration. After concentration, it is temporarily stored in an ultrafiltration temporary storage tank.

[0217] 6. Microfiltration sterilization: The eluent components of the material temporarily stored in the ultrafiltration temporary storage tank in step 5 (i.e., the cation adsorption eluent components and the anion adsorption eluent components) are sterilized through a sterilization membrane with a pore size of 0.6 + 0.2 μm. After sterilization, the material is injected into the aseptic solution preparation system for later use.

[0218] 7. Homogenization: Start the homogenization program, combine the skim milk after step 5 chromatography (secondary chromatography skim milk) with the light cream obtained after centrifugation and skimming in step 2, heat to 55°C, and put into the homogenizer. The homogenization pressure is 240.0 bar to obtain homogenized bovine colostrum.

[0219] 8. Ultra-high temperature instantaneous invasive sterilization: The homogenized bovine colostrum obtained in step 7 is injected into an ultra-high temperature instantaneous invasive sterilization system. The sterilization conditions are 135°C for 1 second. After sterilization, the temperature is reduced to 15°C and injected into an aseptic liquid preparation system to mix with the sterilized material in step 6.

[0220] 9. Aseptic solution preparation: All materials are thoroughly mixed in the aseptic solution preparation system for 5 minutes;

[0221] 10. Aseptic filling: The uniformly mixed materials are fed into the aseptic filling system for commercial aseptic filling;

[0222] 11. Coding and Packing: Coding, packing, and finally warehousing.

[0223] Example 3:

[0224] 1. Acceptance of bovine colostrum: After the frozen bovine colostrum blocks arrive at the factory, samples are taken for testing according to the quality control plan. After arrival, the colostrum blocks are stored in a cold storage at a temperature of -14.5℃. The released bovine colostrum blocks are crushed in a crusher and then put into a dissolving tank. The solid bovine colostrum is thawed and dissolved into liquid by heating with hot water on the tank wall and stirring. After the temperature reaches 12℃, it is pumped out and cooled by a plate heat exchanger before being sent to a storage tank for storage. The storage temperature of bovine colostrum is 7℃.

[0225] 2. Centrifugal defatting: The bovine colostrum stored in the storage tank in step 1 is pumped into the heat recovery section of a three-stage plate heat exchanger. The product, which has been preheated by microfiltration and sterilization in step 3, is then heated to 53°C in the heating section and defatted by a centrifuge (centrifuge speed 4000 r / min) until the fat content is reduced to 0.08%, thus obtaining centrifuged defatted bovine colostrum (skimmed milk) and light cream.

[0226] 3. Microfiltration for oil removal and sterilization: The bovine colostrum obtained in step 2 after centrifugation and defatting is separated by a 1.4μm pore size microfiltration membrane. The material after oil removal and sterilization is returned to the three-stage plate heat recovery section of the centrifugation and defatting process in step 2 to preheat the feed material in step 2. Then, the defatted milk after microfiltration for oil removal and sterilization is cooled to 12°C in the cooling section and stored in a buffer tank for later use.

[0227] 4. Cation chromatography: The skim milk that has been microfiltered and sterilized in the buffer tank of step 3 is pumped into a primary chromatography column via a degassing tank for adsorption chromatography with the cation exchange resin in the column (particle size 220 μm). The temperature of the skim milk is 12℃. The linear flow rate of the skim milk that has been microfiltered and sterilized in step 3 is 700 cm / h, and the column height is 25 cm. After chromatography, the flow-through liquid (i.e., the skim milk after primary chromatography) is collected. Then, it is first flushed with RO water to remove the skim milk from the resin and discharged to the ground. Then, the resin-affinity alkaline proteins such as lactoferrin and peroxidase (cation adsorption eluent components) are eluted with a NaCl solution with a mass fraction of about 5.5%. The eluent is then pumped into a 30 kD ultrafiltration system for desalting and concentration. After concentration, it is temporarily stored in an ultrafiltration temporary storage tank.

[0228] 5. Anion chromatography: The skim milk obtained from the first chromatography in step 4 is pumped into a second chromatography column via a degassing tank for adsorption chromatography with the anion exchange resin in the column (particle size 180 μm). The skim milk temperature is 12℃, the linear flow rate of the skim milk after the first chromatography is 80 cm / h, and the column height is 24 cm. After chromatography, the flow-through liquid (i.e., the skim milk from the second chromatography or the skim milk after chromatography) is collected. Then, it is first flushed with RO water to remove the skim milk from the resin and then drained. Then, the resin-affinity immunoglobulins, bovine serum albumin, etc. (anion adsorption eluent components) are eluted with a NaCl solution of about 5.5% by mass. The eluent is then pumped into a 30 kD ultrafiltration system for desalting and concentration. After concentration, it is temporarily stored in an ultrafiltration storage tank.

[0229] 6. Microfiltration sterilization: The materials temporarily stored in the ultrafiltration temporary storage tank in step 5 (elution components, i.e., cation adsorption eluent components and anion adsorption eluent components) are sterilized through a sterilization membrane with a pore size of 0.6 + 0.2 μm. After sterilization, they are injected into the aseptic solution preparation system for later use.

[0230] 7. Homogenization: Start the homogenization program, combine the skim milk after step 5 chromatography (secondary chromatography skim milk) with the light cream obtained after centrifugation and skimming in step 2, heat to 53°C, and put into the homogenizer. The homogenization pressure is 220 bar to obtain homogenized bovine colostrum.

[0231] 8. Ultra-high temperature instantaneous invasive sterilization: The homogenized bovine colostrum obtained in step 7 is injected into an ultra-high temperature instantaneous invasive sterilization system. The sterilization conditions are 135℃ for 1 second. After sterilization, the temperature is reduced to 12℃ and injected into an aseptic liquid preparation system to mix with the sterilized material in step 6.

[0232] 9. Aseptic solution preparation: All materials are thoroughly mixed in the aseptic solution preparation system for 3 minutes;

[0233] 10. Aseptic filling: The uniformly mixed materials are fed into the aseptic filling system for commercial aseptic filling;

[0234] 11. Coding and Packing: Coding, packing, and finally warehousing.

[0235] Comparative Example 1

[0236] 1. Acceptance of bovine colostrum: After the frozen bovine colostrum blocks arrive at the factory, samples are taken for testing according to the quality control plan. After arrival, the colostrum blocks are stored in a cold storage at a temperature of -14.5℃. The released bovine colostrum blocks are crushed in a crusher and then put into a dissolving tank. The solid bovine colostrum is thawed and dissolved into liquid by heating with hot water on the tank wall and stirring. After the temperature reaches 12℃, it is pumped out and cooled by a plate heat exchanger before being sent to a storage tank for storage. The storage temperature of bovine colostrum is 7℃.

[0237] 2. Centrifugal defatting: The bovine colostrum in the storage tank is pumped into the heat recovery section of a three-stage plate heat exchanger to preheat the defatted and sterilized product. Then it enters the heating section and is heated to 53°C before being defatted by a centrifuge. The fat content is reduced to 0.08%, resulting in centrifuged defatted bovine colostrum (skimmed milk) and light cream.

[0238] 3. Microfiltration for oil removal and sterilization: The bovine colostrum obtained in step 2 after centrifugation and defatting is separated by a 1.4μm pore size microfiltration membrane. The material after oil removal and sterilization is returned to the three-stage plate heat recovery section of the centrifugation and defatting process in step 2 to preheat the feed material in step 2. Then, the defatted milk after microfiltration for oil removal and sterilization is cooled to 12°C in the cooling section and stored in a buffer tank for later use.

[0239] 4. Cation chromatography: The skim milk that has been degreased and sterilized by microfiltration in the buffer tank of step 3 is pumped into a primary chromatography column via a degassing tank for adsorption chromatography with the cation exchange resin in the column (particle size 220um). The temperature of the skim milk is 12℃, the linear flow rate of the skim milk in step 3 is 1200cm / h, and the column height is 25cm. After chromatography, the flow-through liquid (i.e., the skim milk after primary chromatography) is collected. Then, it is first flushed with RO water to remove the skim milk in the resin and discharged to the ground. Then, the resin-affinity alkaline proteins such as lactoferrin and peroxidase (cation adsorption eluent components) are eluted with a NaCl solution with a mass fraction of about 3.0%. The eluent is then pumped into a 30kD ultrafiltration system for desalting and concentration. After concentration, it is temporarily stored in an ultrafiltration temporary storage tank.

[0240] 5. Anion chromatography: The skimmed milk obtained from the first chromatography in step 4 is pumped into a second chromatography column via a degassing tank for adsorption chromatography with the anion exchange resin in the column (particle size 180 μm). The skimmed milk temperature is 12℃, the linear flow rate is 300 cm / h, and the column height is 24 cm. After chromatography, the flow-through liquid (i.e., the skimmed milk from the second chromatography or the skimmed milk after chromatography) is collected. Then, it is first flushed with RO water to remove the skimmed milk from the resin and discharged to the ground. Then, the resin-affinity immunoglobulins, bovine serum albumin, etc. (anion adsorption eluents) are eluted with a NaCl solution with a mass fraction of about 3.0%. The eluent is then pumped into a 30 kD ultrafiltration system for desalting and concentration. After concentration, it is temporarily stored in an ultrafiltration temporary storage tank.

[0241] 6. Microfiltration sterilization: The materials temporarily stored in the ultrafiltration temporary storage tank in step 5 (elution components, i.e., cation adsorption eluent components and anion adsorption eluent components) are sterilized through a sterilization membrane with a pore size of 0.6 + 0.2 μm. After sterilization, they are injected into the aseptic solution preparation system for later use.

[0242] 7. Homogenization: Start the homogenization program, combine the skim milk after step 5 chromatography (secondary chromatography skim milk) with the light cream obtained after centrifugation and skimming in step 2, heat to 53°C, and put into the homogenizer. The homogenization pressure is 220 bar to obtain homogenized bovine colostrum.

[0243] 8. Ultra-high temperature instantaneous invasive sterilization: The homogenized bovine colostrum obtained in step 7 is injected into an ultra-high temperature instantaneous invasive sterilization system. The sterilization conditions are 135℃ for 1 second. After sterilization, the temperature is reduced to 12℃ and injected into an aseptic liquid preparation system to mix with the sterilized material in step 6.

[0244] 9. Aseptic solution preparation: All materials are thoroughly mixed in the aseptic solution preparation system for 3 minutes;

[0245] 10. Aseptic filling: The uniformly mixed materials are fed into the aseptic filling system for commercial aseptic filling;

[0246] 11. Coding and Packing: Coding, packing, and finally warehousing.

[0247] Comparative Example 2:

[0248] 1. Acceptance of bovine colostrum: After the frozen bovine colostrum blocks arrive at the factory, samples are taken for testing according to the quality control plan. After arrival, the colostrum blocks are stored in a cold storage at a temperature of -14.5℃. The released bovine colostrum blocks are crushed in a crusher and then put into a dissolving tank. The solid bovine colostrum is thawed and dissolved into liquid by heating with hot water on the tank wall and stirring. After the temperature reaches 12℃, it is pumped out and cooled by a plate heat exchanger before being sent to a storage tank for storage. The storage temperature of bovine colostrum is 7℃.

[0249] 2. Ultra-high temperature instantaneous invasive sterilization: Bovine colostrum is injected into an ultra-high temperature instantaneous invasive sterilization system. The sterilization conditions are 135℃ for 1 second. After sterilization, the temperature is reduced to 12℃ and injected into a sterile solution preparation system.

[0250] 3. Aseptic filling: The material is fed into the aseptic filling system for commercial aseptic filling;

[0251] 4. Coding and Packing: Coding, packing, and finally warehousing.

[0252] Comparative Example 3: Replicating the technical methods of CN105981908B

[0253] 1. Acceptance of bovine colostrum: After the frozen bovine colostrum blocks arrive at the factory, samples are taken for testing according to the quality control plan. After arrival, the colostrum blocks are stored in a cold storage at a temperature of -14.5℃. The released bovine colostrum blocks are crushed in a crusher and then put into a dissolving tank. The solid bovine colostrum is thawed and dissolved into liquid by heating with hot water on the tank wall and stirring. After the temperature reaches 12℃, it is pumped out and cooled by a plate heat exchanger and sent to a storage tank for storage. The storage temperature of bovine colostrum is 7℃.

[0254] 2. Centrifugal degreasing: The bovine colostrum in the storage tank is pumped into the heat recovery section of a three-stage plate heat exchanger for preheating after degreasing and sterilization. Then it enters the heating section and is heated to 53°C before being degreased by a centrifuge, reducing the fat content to 0.08%.

[0255] 3. Precipitate casein: Adjust the pH of bovine colostrum to 4.5 using acid precipitation to precipitate casein;

[0256] 4. Filtration: Use a 140-mesh filter bag to roughly remove casein and obtain the filtrate;

[0257] 5. Centrifugation: Centrifuge the above filtrate at 4000 rpm for 20 min to remove the precipitate and obtain the supernatant, which contains approximately 15% casein.

[0258] 6. Acidity adjustment: The supernatant obtained in step 5 is acidified with 5% potassium bicarbonate to achieve a pH of 4.5.

[0259] 7. Preparation of a sterile environment: Place the membrane processing equipment and 0.8μm, 0.45μm and 0.2μm polypropylene filter membranes into a clean bench and irradiate with ultraviolet light for 1 hour to achieve a sterile effect;

[0260] 8. Membrane filtration: Add the supernatant with the pH adjusted in step 6 to the membrane filtration device, and let it pass through 0.8μm, 0.45μm and 0.2μm filter membranes in sequence under a membrane pressure difference of about 2 Bar. Then store the solution in a sterile sealed bag to obtain a sterile liquid bovine colostrum solution.

[0261] Test Example 1: Validation of Anion Chromatography Process

[0262] The skim milk after chromatography was detected using a flow-through UV detector. Figures 2A-2D Chromatographic chromatograms of the examples and comparative examples are shown, and based on... Figures 3A-3D The results analysis shows that changes in core process parameters have a significant impact on the preparation process. In Comparative Example 1, the immunoglobulin extraction process could not proceed normally due to changes in process conditions.

[0263] Test Example 2: Commercial Sterility Testing

[0264] According to the relevant requirements of "Low-acid foods, acidified foods - dairy products, beverages and other liquid foods" in GB 4789.26-2023 National Food Safety Standard for Microbiological Examination of Foods - Commercial Sterility Examination, specific examples were tested, and the specific results are shown in Table 1 below:

[0265] Table 1:

[0266]

[0267] Based on the test results, all specific embodiments achieved commercial sterility and met the relevant requirements of GB 4789.26-2023 National Food Safety Standard for Microbiological Examination of Food - Commercial Sterility Test.

[0268] Test Example 3: Comparison of Nutritional Content

[0269] Based on the following standards: GB / T 5009.194-2003 Determination of Immunoglobulin IgG in Health Foods, GB 5009.299-2024 National Food Safety Standard Determination of Lactoferrin in Foods, GB 5009.5-2025 National Food Safety Standard Determination of Protein in Foods, GB 5009.6-2025 National Food Safety Standard Determination of Fat in Foods, QFH0019S—2025 Milk Fat Globule Membrane Protein Powder, GB 5009.14-2017 National Food Safety Standard Determination of Zinc in Foods, GB 5009.90-2016 National Food Safety Standard Determination of Iron in Foods, GB 5009.82-2016 National Food Safety Standard Determination of Vitamins A, D, and E in Foods, and GB 5009.285-2022... The specific examples and comparative examples were analyzed and compared using methods such as the "National Food Safety Standard for the Determination of Vitamin B12 in Food". The specific results are shown in Table 2 below:

[0270] Table 2:

[0271]

[0272] Analysis based on the test results in Table 2:

[0273] The contents of fat, phospholipids, sphingomyelin, and gangliosides in each embodiment were significantly higher than those in Comparative Example 3, indicating that bovine colostrum whole milk, compared with Comparative Example 3, removed milk fat and casein micelles and retained a more complete fat globule membrane (MFGM) system.

[0274] Regarding immunoglobulin distribution, the whole colostrum in each example exhibited stable triple spatial anchoring in the "micelle phase-whey phase-lipid phase," while Comparative Example 3 contained only whey phase IgG. The total IgG content in bovine colostrum was 5-6%, with a large amount of IgG adsorbed onto the surface of casein micelles via electrostatic interactions, and another portion bound to the phospholipid head group of MFGM. Although 0.1 μm microfiltration allowed monomeric IgG (hydrated diameter ~7 nm) to pass through freely, it completely retained both micellar-bound and MFGM-bound IgG. Therefore, although the actual IgG concentration in the permeate could reach a higher percentage of 6-7% after ultrafiltration concentration, the overall yield was reduced by at least 35% compared to whole milk.

[0275] Regarding the maintenance of the bioactivity of growth factors and signaling molecules, it can be seen that, compared with Comparative Example 3 which removed milk fat and casein micelles, the contents of insulin-like growth factor-I (IGF-I) and transforming growth factor-β (TGF-β) in whole colostrum of each example are clearly lipid microenvironment dependent and are higher.

[0276] Regarding trace elements, it can be seen that, compared with Comparative Example 3 which removed milk fat and casein micelles, the trace element content in the whole colostrum of each embodiment was higher and the loss was lower.

[0277] Compared with Comparative Example 1, the preparation method provided by the present invention significantly increases the content of core indicators such as immunoglobulins and lactoferrin in whole colostrum. Specifically, the preparation method provided by the present invention has a dry matter content greater than 15g / 100g, a protein content greater than 9g / 100g, an immunoglobulin (IgG) content greater than 5%, a lactoferrin (LF) content greater than 50mg / 100g, and a ganglioside content greater than 9mg / 100g.

[0278] Compared with Comparative Example 2, the ultra-high temperature instantaneous invasive sterilization method in each embodiment, while having certain advantages in preserving the nutrients of whole colostrum compared to traditional thermal sterilization processes, resulted in significant losses of immunoglobulins and lactoferrin. In summary, the preparation method provided by this invention effectively preserves the content of nutrients such as immunoglobulins and lactoferrin in bovine colostrum.

[0279] It should be noted that although the technical solution of the present invention has been described with specific examples, those skilled in the art will understand that the present invention should not be limited thereto.

[0280] The various embodiments of the present invention have been described above. These descriptions are exemplary and not exhaustive, nor are they limited to the disclosed embodiments. Many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the described embodiments. The terminology used herein is chosen to best explain the principles, practical application, or technical improvements to the embodiments in the market, or to enable others skilled in the art to understand the embodiments disclosed herein.

Claims

1. A method for preparing a liquid promulgation, characterized in that, The method includes: The defatting process involves separating the dairy raw materials to obtain skim milk and cream. The adsorption chromatography step involves treating the skim milk with ion exchange chromatography to obtain chromatographically purified skim milk and eluent components, wherein the eluent used in the ion exchange chromatography has a salt mass fraction greater than 3%. The homogenization step involves homogenizing the skim milk after chromatography with the light cream to obtain a homogenized material. The mixing step involves mixing the homogenized material with the elution components to obtain the liquid colostrum; Optionally, the colostrum includes at least one of bovine colostrum, sheep colostrum, camel colostrum, and horse colostrum.

2. The method according to claim 1, characterized in that, In the adsorption chromatography step, the skim milk is subjected to cation exchange chromatography to obtain a flow-through liquid, and eluted with an eluent to obtain the cationic adsorbed eluent components, wherein the linear flow rate of the skim milk in the cation exchange chromatography is less than 1200 cm / h.

3. The method according to claim 1 or 2, characterized in that, The cation adsorption and elution components include basic proteins; Optionally, the basic protein includes at least one of lactoferrin, β-lactoglobulin, bovine serum albumin, and peroxidase.

4. The method according to claim 2 or 3, characterized in that, In the adsorption chromatography step, the flow-through liquid obtained by the cation exchange chromatography is subjected to anion exchange chromatography to obtain the chromatographically defatted emulsion, and the anion adsorption eluent is used to elute the anion adsorbed eluent components, wherein the linear flow rate of the flow-through liquid in the anion exchange chromatography is less than 300 cm / h.

5. The method according to any one of claims 1-4, characterized in that, The anion adsorption and elution components include nutrients; Optionally, the nutrients include immunoglobulins and / or bovine serum albumin.

6. The method according to any one of claims 1-5, characterized in that, In the adsorption chromatography step, the temperature of the skim milk is below 25°C; Preferably, the temperature of the skim milk is below 18°C.

7. The method according to any one of claims 1-6, characterized in that, Prior to the homogenization step, the method further includes an ultrafiltration step for the eluted components; Preferably, the ultrafiltration process includes solution displacement and / or concentration of the eluted components.

8. The method according to any one of claims 1-7, characterized in that, The method further includes a sterilization process, wherein the sterilization process includes sterilizing skim milk, ultrafiltration material, and / or homogenized material.

9. A liquid promulgation prepared by the preparation method according to any one of claims 1-8.

10. A formula dairy product, characterized in that, The formulated dairy product includes: liquid colostrum prepared by the preparation method according to any one of claims 1-8 or liquid colostrum as described in claim 9. Optionally, the formulated dairy product may also contain one or more of the following additional components: (A) Functional protein components, (B) Carbohydrate components, (C) Fats and oils, (D) Mineral salts, (E) Other nutritional supplement components.

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