Lithocholic acid additive for feline obesity or metabolic syndrome management, method of preparation and use

Abstract

The present application relates to pet food science and nutrition and health care technical field, specifically disclose a kind of for cat obesity or metabolic syndrome management LCA additive, preparation method and application, by weight parts include, LCA 5~15 parts, maltodextrin 70~88 parts, microcrystalline cellulose 5~20 parts, taste masking agent 1~5 parts, silicon dioxide 0.2~1.0 parts, tocopherol 0.05~0.2 parts;LCA, maltodextrin and microcrystalline cellulose are premixed, and basic premix C is obtained;Chicken liver powder is added in basic premix C and mixed, then tocopherol is added and mixed, and then silicon dioxide is sprinkled and mixed, and LCA additive powder is obtained;Fish hydrolysate and yeast extract are dissolved in sunflower oil to obtain oil phase solubles;Oil phase solubles are added to LCA additive powder by spraying while stirring to obtain LCA additive liquid.The present application has the characteristics of good effect on improving the body weight of obese cats, good improvement in glucose and blood lipid metabolism and multiple health benefits.

Description

Technical Field

[0001] This invention belongs to the field of pet food science and nutrition technology, specifically relating to a lithocholic acid additive for the management of obesity or metabolic syndrome in cats, its preparation method, and its application. Background Technology

[0002] With the increasing prevalence of pets in households and changes in urban lifestyles, overweight and obesity in domestic cats are on the rise and are significantly associated with risks such as insulin resistance, glucose and lipid metabolism disorders, hepatobiliary and pancreatic diseases, arthritis, and shortened lifespan. Multiple reviews and national / regional cohorts indicate that the overall prevalence of obesity or overweight in cats is commonly 30%-40%, exceeding 50% in some regions; recent studies have also reported rates as high as 63% in New Zealand and nearly 40% in the United States, reflecting the significant burden of obesity in cats. Current pet obesity interventions are based on energy-restricted prescription feeding, behavioral modification, and exercise management, but long-term adherence and sustainability are insufficient. Drug and surgical interventions in the pet field are limited in clinical application due to safety, applicability, and cost-effectiveness. In contrast, nutritional interventions that can be integrated into daily life, such as dietary therapy, are considered a more accessible pathway, focusing on long-term, gentle, and sustainable regulation of body weight and metabolic profiles by targeting gut-bile acid metabolism and signaling. In this technical approach, bile acids play a particularly prominent role as metabolic signaling molecules. Bile acids can activate the membrane receptor TGR5 (GPBAR1), promoting the release of GLP-1 from intestinal L cells, thereby affecting energy expenditure and glucose and lipid metabolism. Lithocholic acid (LCA) has been repeatedly proven to be one of the stronger natural TGR5 agonists, providing a clear molecular approach and an operable dosimetric basis for nutritional intervention. Adding LCA directly to a cat's daily diet based on body weight can be implemented in a home setting without changing prescription food, and is expected to achieve steady improvements in indicators such as body weight, BCS, FBMI, abdominal circumference, GLU, TG, TC, LDL, and HDL.

[0003] In recent years, the number of dogs and cats and pet health expenditures have continued to grow, with obesity becoming a core issue in pet health management. The market favors sustainable, easily integrated, and highly adherent nutritional solutions. Unlike one-off, intensive interventions, standardized dietary additives can achieve long-term adherence through weight-based measurements and home mixing, facilitating standardized packaging and labeling, such as weight-based bagging. Industry surveys also show a significant gap between public awareness of pet obesity and actual prevalence, suggesting the practical value of low-barrier and long-term implementable management tools. The interaction between gut microbiota and bile acids in the canine and feline gut has been widely studied. The conversion of primary bile acids to secondary bile acids is highly dependent on microbial metabolism. Peptacetobacter hiranonis has been shown to be significantly associated with the bai gene cluster and secondary bile acid levels, serving as a key functional hub for maintaining fecal unconjugated bile acid and bile acid homeostasis. In feline data, the correlation between C. hiranonis abundance and unconjugated bile acid levels, along with their temporal variability, has also been observed, providing biological evidence for the microbial transformation of LCAs and the synergistic effects of their products.

[0004] It is worth emphasizing that LCAs can be further modified by gut microbiota, such as through oxidation, reduction, isomerization, and conjugation, to generate a group of LCAs and their derivatives with immunometabolic activity, including isoLCA, 3-oxoLCA, 7-ketoLCA, 12-ketoLCA, isoalloLCA, tauroLCA, and glycoLCA. Numerous studies have confirmed the bioactivity of LCAs derived from gut microbiota. 3-oxoLCA inhibits Th17 differentiation by directly binding to RORγt; isoalloLCA promotes Treg differentiation through the mitochondrial ROS-FOXP3 axis; gavage supplementation of mice with 3-oxoLCA or isoalloLCA can reduce Th17 and increase Treg levels in the intestinal lamina propria; these effects provide evidence for the immunometabolic linkage mechanism in improving body weight and metabolic profile. It is worth mentioning that, regarding safety boundaries and control treatments, in rodent models, short-term high exposure to LCA can induce cholestasis and liver damage, suggesting that in pet applications, it is essential to adhere to body weight-based dosing and conduct dynamic monitoring of ALT or AST, as well as individualized downregulation or discontinuation strategies to ensure the safety and sustainability of long-term use. On the other hand, ursodeoxycholic acid (UDCA), commonly used in clinical veterinary medicine, is mainly used for the management of hepatobiliary diseases in cats, such as cholestasis, cholangitis, or cholelithiasis. The common oral dose is approximately 15-25 mg / kg / day. Its indications and mechanisms of action differ from those of weight loss or metabolic management, and it can serve as a control background for usage and technical approaches. Based on the above evidence, a weight-based LCA diet supplementation regimen, combined with follow-up of fecal LCA, gut microbial derivatives, and metabolic indicators, is expected to achieve: quantifiable improvements in weight, BCS, FBMI, waist circumference, GLU, TG, TC, LDL, and HDL; changes in the levels of fecal LCA and gut microbial derivatives: Lithocholic acid, 7-Ketolithocholic acid, 12-Ketolithocholic acid, Isolithocholic acid, Taurolithocholic acid, Dehydrolithocholic acid, Glycolithocholic acid, or Isoallolithocholic acid; long-term sustainability within an ALT or AST-gated framework; and standardized premixes and quantitative sachets labeled by weight, supporting daily implementation and adherence in family settings.

[0005] With the increasing prevalence of pets in households and changes in urban lifestyles, overweight or obesity in domestic cats is becoming increasingly prominent, significantly associated with health risks such as insulin resistance or disorders of glucose and lipid metabolism. Existing interventions, such as energy-restricted prescription feeding, medication, and surgery, suffer from problems such as insufficient long-term adherence, limited safety, and limited applicability. Regulating intestinal bile acid metabolism and signaling pathways through nutritional intervention is considered a more accessible management approach. Bile acids, as metabolic signaling molecules, especially lithocholic acid (LCA) as a natural TGR5 agonist, have clear potential in regulating energy metabolism; however, current nutritional intervention techniques still have shortcomings. For example, patent document CN120937998A discloses a microcapsule flavoring powder for cats containing active plant ingredients and its preparation method. The powder includes a double-layer encapsulation system. By weight percentage, the raw material components are: 10-14 wt% composite core material, 32-36 wt% multifunctional palatability-enhancing base material, 35-39 wt% double-layer encapsulation system, and 14-16 wt% functional excipients. The composite core material is a mixture of peppermint extract, green tea extract, chicory extract, mulberry leaf extract, and hawthorn extract in a mass ratio of (1.8-4.5):(1.2-3):(0.8-1.5):(0.5-1.2):(1-2). This method achieves a dual-stage targeting effect of low release in the stomach and high release in the intestines, while also possessing high palatability, promoting intestinal health, improving metabolism, and synergistic anti-aging effects. It is also long-lasting and stable, and can be adapted to various types of functional cat foods. However, this method is still not very effective in reducing the weight of obese cats and improving blood sugar and blood lipids.

[0006] Therefore, existing products for improving feline obesity have problems such as insufficient effect on improving the weight of obese cats, inadequate improvement in glucose and lipid metabolism, and limited health benefits. Summary of the Invention

[0007] In order to solve the above-mentioned technical problems of existing products for improving feline obesity, the present invention provides a lithocholic acid additive, preparation method and application for the management of feline obesity or metabolic syndrome. It has the characteristics of good effect on improving the weight of obese cats, good effect on improving glucose and lipid metabolism and multiple health benefits.

[0008] The first technical solution of the present invention: a lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following components by weight:

[0009] The ingredients are: 5-15 parts lithocholic acid, 70-88 parts maltodextrin, 5-20 parts microcrystalline cellulose, 1-5 parts flavor masking agent, 0.2-1.0 parts silica, and 0.05-0.2 parts tocopherol. Lithocholic acid (LCA) is the main functional component in this invention. It can promote GLP-1 secretion by activating the intestinal TGR5 receptor, thereby improving energy metabolism and blood glucose and lipid levels, and regulating obesity and metabolic syndrome in cats. Maltodextrin, as the main diluent, can adjust the lithocholic acid content to a suitable dietary dosage range and ensure measurement accuracy during packaging. It also has a certain flavor masking effect, which is beneficial for long-term feeding. Microcrystalline cellulose (MCC), as a safe and inert carrier, has good adsorption and dilution effects, ensuring the uniform distribution of lithocholic acid in the additive and improving overall flowability and processing performance. The flavor masking agent has a significant flavor-improving effect and can mask the flavor of lithocholic acid. Bitterness and unpleasant odor enhance cats' feeding compliance and ensure the sustainability of intervention effects; silica, as an adjuvant, acts as an anti-caking agent, reducing powder aggregation during storage and packaging, maintaining the flowability and stability of the premix, and facilitating industrial production; tocopherol, as an adjuvant, acts as an antioxidant, effectively inhibiting the oxidative degradation of lithocholic acid and lipid components in the formulation, improving formulation stability, and extending product shelf life; this invention provides a basic formulation framework to ensure uniform dispersion, good flowability, acceptable palatability, and antioxidant stability of lithocholic acid LCA, wherein a limited proportion of microcrystalline cellulose and maltodextrin serve as an inert framework to carry and dilute lithocholic acid LCA, thus... The method involves weighing cats and using a limited number of masking agents to cover the bitterness and irritation of lithocholic acid (LCA). A stepwise mixing process is employed to achieve a uniform distribution of low-content active ingredients, ultimately producing an additive with controllable content and intra-batch uniformity (RSD ≤ 10%). This allows for selective and precise feeding at 15–45 mg / (kg body weight·day) based on the cat's specific tolerance. This invention can be added to prescription diets as needed, eliminating the need to change the diet and facilitating home use. It effectively reduces weight, abdominal circumference, body condition score (BCS), and body fat index (FBMI) in obese cats; it also improves blood glucose and lipids in obese cats, reducing triglycerides (TG), total cholesterol (TC), and low-density lipoprotein cholesterol (LDL). It reduces certain indicators; enhances antioxidant capacity, increasing glutathione peroxidase (GSH-Px) and decreasing malondialdehyde (MDA) levels in obese cats; strengthens humoral immunity in obese cats, increasing immunoglobulin AIg, immunoglobulin GIgG, and immunoglobulin MIgM; and improves low-grade inflammation in obese cats, decreasing pro-inflammatory cytokines such as interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α), while increasing the anti-inflammatory cytokine interleukin-10 (IL-10). It exhibits multiple health benefits, including significantly improving weight, glucose and lipid metabolism, and overall health in obese cats.

[0010] Preferably, the product comprises the following components in parts by weight.

[0011] Lithocholic acid 8-12 parts, maltodextrin 75-85 parts, microcrystalline cellulose 8-18 parts, flavor masking agent 2-4 parts, silica 0.4-0.8 parts, tocopherol 0.08-0.18 parts. This formula narrows the formulation range, improving process robustness and stabilizing the lithocholic acid (LCA) content at approximately 10%, with about 100 mg of LCA per gram. This facilitates dosage conversion, reduces batch-to-batch variability, and enhances mixing uniformity. The proportions of microcrystalline cellulose and maltodextrin as inert frameworks are closer to the actual mixing limits, reducing the risk of stratification.

[0012] Preferably, the product comprises the following components in parts by weight.

[0013] Lithocholic acid 9-11 parts, maltodextrin 78-82 parts, microcrystalline cellulose 10-15 parts, flavor masking agent 2-4 parts, silica 0.5-0.7 parts, tocopherol 0.1-0.15 parts. The formulation was further optimized to improve flowability and flavor masking effect. The slightly increased proportion of microcrystalline cellulose (MCC) resulted in better flowability and disintegration, enhancing tableting and flowability, making it more suitable for automated packaging lines. The flavor masking agent provides more complete coverage, leading to higher cat acceptance and suitability for mass production, balancing process and palatability.

[0014] Preferably, the product comprises the following components in parts by weight.

[0015] The additive contains 10 parts lithocholic acid, 80 parts maltodextrin, 12 parts microcrystalline cellulose, 3 parts flavor masking agent, 0.6 parts silica, and 0.12 parts tocopherol. Each gram of the additive contains approximately 100 mg of lithocholic acid (LCA). A daily dose of 0.3 g is recommended for a 5 kg cat. The additive is designed for easy small-packaging, offering better uniformity, stability, palatability, and antioxidant properties. All excipients are balanced, with sufficient carrier dilution, adequate anti-caking properties, sufficient flavor masking, and satisfactory antioxidant performance.

[0016] Preferably, the product also includes the following components by weight: 90-100 parts sunflower seed oil, 1-2 parts fish hydrolysate, and 1-2 parts yeast extract. This forms a liquid or oil-based additive, allowing lithocholic acid (LCA) to dissolve in the oil phase and disperse better, resulting in a liquid additive suitable for wet food, sauce packets, or post-coating. This improves palatability, making it suitable for cats that don't like powdered food, or for surface coating of dry food. The tocopherol component also protects against oil oxidation, creating a differentiated formulation that covers more application scenarios.

[0017] Preferably, the additive also includes the following components by weight: 92-98 parts sunflower seed oil, 1.2-1.8 parts fish hydrolysate, and 1.2-1.8 parts yeast extract. Narrowing the oil phase ratio ensures stable additive concentration, controls the LCA content per milliliter of additive, facilitates metering with droppers or pump heads, improves dosage accuracy, and reduces content deviations caused by oil volume fluctuations.

[0018] Preferably, the additive also includes the following components by weight: 95 parts sunflower seed oil, 1.5 parts fish hydrolysate, and 1.5 parts yeast extract. Each milliliter of the additive provides approximately 20–30 mg of lithocholic acid (LCA), which is convenient to add dropwise according to the cat's body weight, such as 1 mL / 5 kg cat. The 95% oil phase provides good flowability, oxidative stability, and palatability.

[0019] Preferably, the flavor masking agent is selected from at least one of chicken liver powder, fish hydrolysate, or yeast extract. It can mask the bitterness of lithocholic acid (LCA) and the distinctive odor of bile acids, increasing the cat's willingness to eat and reducing behaviors such as refusal to eat or vomiting. The animal-derived flavor substances compete with bitter taste receptors for binding, or mask the taste through olfaction. All three are natural palatability enhancers in pet food, and cats have a high acceptance rate for them.

[0020] This invention provides an additive with 10% lithocholic acid (LCA) content, which is easy to convert to 30 mg / kg body weight, or 0.3 g / 5 kg cat. The preparation process adopts a stepwise mixing method, which can achieve accurate content control of each component and meet the requirements of intra-batch uniformity RSD ≤ 10%. Maltodextrin, microcrystalline cellulose (MCC), silica, and tocopherol are all food and feed additives approved by the Food and Agriculture Organization of the United Nations (FAO), the WHO, or the US Food and Drug Administration (FDA). The masking agent can effectively cover the unpleasant flavor of lithocholic acid (LCA), the sunflower seed oil phase can further improve acceptability, and the tocopherol can resist oxidation and prevent oxidative rancidity of the oil phase. It can be prepared as a solid or liquid as needed, with various dosage forms to choose from, and can cover dry food, wet food, sauce packets, and post-spraying.

[0021] Preferably, the lithocholic acid is obtained by enzymatically dehydroxylating ursodeoxycholic acid (UDCA) as a starting substrate. This clarifies the production route of lithocholic acid LCA as a bio-enzymatic conversion, replacing chemical synthesis or animal bile extraction, thus improving product specificity and ultimately yielding high-purity lithocholic acid LCA with low byproducts. Ursodeoxycholic acid (UDCA) differs from lithocholic acid LCA only by a 12α-hydroxyl group; selective removal of this hydroxyl group yields lithocholic acid LCA. UDCA is relatively readily available, as it can be converted from deoxycholic acid, and the enzymatic dehydroxylation method is highly efficient and environmentally friendly.

[0022] Preferably, the enzymatic directional dehydroxylation reaction of lithocholic acid includes the following steps:

[0023] (a01) Enzyme system preparation

[0024] (a01-1) Clostridium hiranonis strain was inoculated into enhanced Clostridium culture medium for anaerobic culture; the enhanced Clostridium culture medium was a commercial standard Clostridium-specific culture medium, and the anaerobic culture environment was an anaerobic atmosphere of 95% N2 and 5% CO2, and cultured in an anaerobic fermenter or anaerobic workstation.

[0025] (a01-2) Collect the bacterial cells by centrifugation of the culture medium in step (a01-1), add an appropriate amount of PBS buffer to resuspend, and then sonicate and centrifuge to obtain the supernatant. Sonicate the cells under the conditions of 200W power, 5s working time, 5s rest time, and a total time of 15min. Centrifuge at 12000rpm for 20min to obtain the supernatant and obtain the crude enzyme solution.

[0026] (a01-3) Add ammonium sulfate to the crude enzyme solution in step (a01-2) for fractional precipitation, and collect the precipitate with a saturation of 30% to 60%;

[0027] (a01-4) Dissolve the precipitate in step (a01-3) with sterile water, then purify by DEAE-Sepharose FastFlow ion exchange chromatography, collect the elution peak, and take the fractions from each tube to detect the 12α-hydroxy removal activity as follows: take 100 μL of each fraction, add 1 mL of reaction solution containing 10 mg / mL UDCA, incubate at 37℃ for 30 min, and then detect the LCA production by HPLC. Combine the fractions with the highest activity, freeze dry, and obtain enzyme powder;

[0028] (a02) Enzymatic reaction

[0029] (a02-1) Dissolve ursodeoxycholic acid (UDCA) with a purity ≥98% in a mixed solution of phosphate buffer and methanol to obtain a substrate solution;

[0030] (a02-2) Add enzyme powder to the substrate solution in step (a02-1) and stir to react;

[0031] (a03) Product purification

[0032] (a03-1) After the reaction in step (a02-2), add an equal volume of ethyl acetate to the mixture and extract multiple times. Combine the organic phases and evaporate to dryness.

[0033] (a03-2) Dissolve the evaporated product from step (a03-1) in hot ethanol and perform hot filtration, then allow the filtrate to stand and age.

[0034] (a03-3) The white crystals obtained from the aging process in step (a03-2) are filtered, washed, and then vacuum dried to obtain lithocholic acid (LCA). The *Clostridium hiranonis* strain is a known 12α-dehydroxylating bacterium, and the enzyme system selection is correct and biologically sound. The process, from strain cultivation, crude enzyme preparation, ion exchange purification, enzymatic conversion, extraction, crystallization to drying, demonstrates good feasibility. Using ursodeoxycholic acid (UDCA) as a starting material keeps costs under control, allows for batch enzyme preparation, and enables solvent recovery, resulting in good economic efficiency.

[0035] Preferably, the Clostridium hiranonis strain has the accession number JCM16549. Clostridium hiranonis provides an enzyme source with 7α- or 12α-hydroxy dehydroxylation activity, ensuring enzyme specificity and activity. It efficiently catalyzes the conversion of ursodeoxycholic acid (UDCA) to lithocholic acid (LCA) with fewer side reactions. Clostridium hiranonis is a known 7α- or 12α-hydroxylating bacterium containing the bai gene cluster. This strain is a model strain, publicly available, and its activity has been verified. The JCM16549 strain is publicly available through the RIKEN Microbial Collection Center (JCM) in Japan and can be obtained through regular commercial channels without special approval.

[0036] Preferably, the culture temperature in step (a01-1) is 35℃~38℃, and the culture time is 36h~60h. More preferably, the culture temperature is 36℃~37℃, and the culture time is 42h~54h. Even more preferably, the culture temperature is 37℃, and the culture time is 48h. Determining the optimal conditions for bacterial growth and enzyme induction maximizes enzyme activity per unit volume, resulting in high-density and highly active bacterial cells. 37℃ is the optimal growth temperature for Clostridium, and 48h is sufficient to reach the stationary phase and peak enzyme activity.

[0037] Preferably, in step (a01-4), the specific activity of the enzyme powder is not less than 15 U / mg protein. Setting a lower limit for enzyme powder quality ensures consistent catalytic efficiency for each batch, reduces enzyme powder usage, controls costs, and establishes a conversion rate of 1 U = 1 μmol / min. 15 U / mg is the standard for industrial-grade enzyme preparations.

[0038] Preferably, the concentration of the substrate solution in step (a02-1) is 50 mg / mL. The initial concentration of ursodeoxycholic acid (UDCA) in the reaction system is determined to balance solubility and conversion efficiency, avoid substrate inhibition, and maintain a high conversion rate. The solubility of UDCA in phosphate buffer and methanol is approximately 50 mg / mL; concentrations higher than this may lead to precipitation or inhibit enzyme activity.

[0039] Preferably, the pH of the mixed solution in step (a02-1) is 6.6–6.9, and the volume ratio of phosphate buffer to methanol in the mixed solution is 3.5–4.5:1. More preferably, the pH of the mixed solution in step (a02-1) is 6.7–6.8, and the volume ratio of phosphate buffer to methanol in the mixed solution is 3.8–4.2:1. This provides an optimal microenvironment for the enzyme reaction, maintains enzyme activity and substrate solubility, resulting in higher conversion rates and fewer byproducts. Methanol helps dissolve ursodeoxycholic acid (UDCA), and excessively high methanol concentrations can inactivate the enzyme. A pH of 6.8 is also close to the optimal pH for clostridium enzymes.

[0040] Preferably, in step (a02-2), the enzyme powder added is 6% to 10% of the substrate solution mass. More preferably, the enzyme powder added is 7% to 9% of the substrate solution mass. Even more preferably, the enzyme powder added is 8% of the substrate solution mass. The economical enzyme dosage should be determined to ensure the reaction is completed within a reasonable time, with a conversion rate ≥95% after 28 to 30 hours. Too little enzyme will result in a slow reaction, while too much enzyme will lead to waste and may introduce impurities.

[0041] Preferably, the reaction temperature in step (a02-2) is 36℃~37℃, the reaction time is 28h~30h, and the pH of the reaction system is 6.7~6.9. More preferably, the reaction temperature in step (a02-2) is 36.5℃, the reaction time is 29h, and the pH of the reaction system is 6.8. Precise control of the reaction conditions achieves the highest conversion rate and the lowest byproducts, ensuring that the conversion rate of ursodeoxycholic acid (UDCA) to lithocholic acid (LCA) is ≥95%. The optimal temperature for this enzyme system is 36.5℃, the optimal pH is 6.8, and the reaction can be completed in 29 hours.

[0042] Preferably, the reaction in step (a02-2) is carried out in a isothermal oscillating reactor. The isothermal oscillating reactor provides uniform mixing, oxygen and mass transfer, prevents substrate or enzyme precipitation, improves reaction uniformity and conversion rate, and the oscillation enhances solid-liquid contact.

[0043] Preferably, the extraction in step (a03-1) is performed 2 to 5 times, and the evaporation is rotary evaporation. More preferably, the extraction is performed 3 to 4 times. This method efficiently recovers lithocholic acid (LCA) from the aqueous phase, reduces product loss, and achieves a recovery rate of ≥90%. Lithocholic acid (LCA) is soluble in ethyl acetate, and rotary evaporation is relatively gentle and rapid.

[0044] Preferably, in step (a03-2), the volume of hot ethanol is 9 to 11 times the volume of the evaporated product. More preferably, the volume of hot ethanol is 10 times the volume of the evaporated product. Crude lithocholic acid (LCA) is dissolved for recrystallization, controlling the crystallization yield and purity to ensure a crystallization yield ≥85% and a purity ≥95%. Hot ethanol has high solubility, and upon cooling, lithocholic acid (LCA) crystals precipitate out.

[0045] Preferably, in step (a03-2), the static aging temperature is 3℃~6℃, and the static aging time is 6h~18h. More preferably, in step (a03-2), the static aging temperature is 4℃~5℃, and the static aging time is 9h~15h. Even more preferably, in step (a03-2), the static aging temperature is 4.5℃, and the static aging time is 12h. This promotes crystal growth and purification, improves crystal purity and particle size uniformity, ensures purity ≥95%, good crystal form, and the low-temperature slow crystallization is beneficial for impurity removal.

[0046] Preferably, the hot ethanol in step (a03-2) is 95% hot ethanol. 95% hot ethanol provides a recrystallization solvent, dissolves lithocholic acid (LCA), removes insoluble impurities by filtration, and achieves the required standard in one step of crystallization. 95% ethanol has high solubility for LCA and low solubility for polar impurities.

[0047] Preferably, the vacuum drying temperature in step (a03-3) is 35℃~45℃, the vacuum drying pressure is -0.1MPa~-0.09MPa, and the vacuum drying time is 3h~9h. More preferably, the vacuum drying temperature in step (a03-3) is 38℃~42℃, the vacuum drying pressure is -0.095MPa, and the vacuum drying time is 4h~8h. Even more preferably, the vacuum drying temperature in step (a03-3) is 40℃, the vacuum drying pressure is -0.095MPa, and the vacuum drying time is 6h. Residual ethanol and moisture are removed to obtain dry, stable, and easily pulverized lithocholic acid (LCA), ensuring moisture content ≤1% and residual ethanol ≤5000ppm. Low-temperature vacuum drying avoids thermal degradation.

[0048] Preferably, the purity of lithocholic acid (LCA) in step (a03-3) is ≥95%. Setting product quality standards ensures the safety and effectiveness of subsequent additives; 95% purity is sufficient as a raw material for feed additives.

[0049] Preferably, the preparation of the microcrystalline cellulose (MCC) includes the following steps:

[0050] (b01) Take bleached wood pulp with α-cellulose content ≥90%, dry and then crush it; bleached wood pulp with α-cellulose content ≥90% provides high-purity cellulose raw materials, ensuring the crystallinity and purity of the final MCC. After acid hydrolysis, there are fewer impurities and higher whiteness. α-cellulose is a long-chain glucose polymer. Acid hydrolysis removes the amorphous region and retains the crystalline region.

[0051] (b02) After adding hydrochloric acid solution to the pulverized material in step (b01) and stirring the reaction, quickly cool it to room temperature with ice water. Quickly cooling to room temperature with ice water can quickly terminate the acid hydrolysis reaction, prevent excessive hydrolysis, and control the degree of polymerization within the target range. The hydrolysis reaction rate will decrease sharply as the temperature decreases. Ice water cooling is an effective means of rapid cooling.

[0052] (b03) After filtering the cooling mixture from step (b02), take the filtrate and wash it with deionized water until the pH of the filtrate is 6.5-7;

[0053] (b04) Dry the filtrate after washing in step (b03) until the moisture content is ≤5%;

[0054] (b05) The dried material from step (b04) is pulverized by an air jet mill and then sieved to obtain microcrystalline cellulose (MCC) powder. Bleached wood pulp is a bulk industrial raw material, and α-cellulose ≥90% is readily available, demonstrating good raw material availability. The process of acid hydrolysis, washing, drying, and air jet milling has good process feasibility. The equipment requires conventional chemical or pharmaceutical equipment and does not require special customization. The reaction temperature, time, solid-liquid ratio, pH, and particle size can all be detected online or offline, enabling good quality control. The prepared microcrystalline cellulose (MCC) can be directly used in the formulation of lithocholic acid (LCA) additives, synergistically acting as a carrier with maltodextrin.

[0055] Preferably, in step (b01), the fibers are pulverized to a length of 1mm to 2mm. This increases the contact area between the acid and cellulose, improves the uniformity and efficiency of hydrolysis, shortens the reaction time, reduces local over-acidity, and ensures good suspension properties in the solid-liquid reaction, making them less prone to entanglement. Fibers that are too long will result in uneven mixing, while fibers that are too fine will generate dust.

[0056] Preferably, in step (b02), the solid-liquid ratio of the pulverized material to the hydrochloric acid solution is 1:10 to 20. More preferably, the solid-liquid ratio is 1:12 to 18. Even more preferably, the solid-liquid ratio is 1:15. The ratio of acid to cellulose is determined to ensure an excess of acid, fully hydrolyzing the amorphous region and reducing the degree of polymerization to 200-400. A solid-liquid ratio that is too low will waste acid, while a ratio that is too high will result in incomplete hydrolysis.

[0057] Preferably, the concentration of the hydrochloric acid solution in step (b02) is 2N. Providing a fixed concentration of hydrolytic acid controls the hydrolysis rate and extent, selectively hydrolyzing the amorphous region without damaging the crystalline region. A 2N concentration of HCl provides a suitable hydrolysis rate for the amorphous region of cellulose at 80°C; concentrations below 2N result in slower hydrolysis, while concentrations above 2N may damage the crystalline region.

[0058] Preferably, the reaction temperature in step (b02) is 80℃~85℃, and the reaction time is 80min~120min. More preferably, the reaction temperature is 81℃~84℃, and the reaction time is 90min~110min. Even more preferably, the reaction temperature is 82℃~83℃, and the reaction time is 95min~105min. Precise control of the degree of hydrolysis is crucial to achieving the target degree of polymerization DP200~400, resulting in a higher yield and better crystallinity. Appropriately increasing the temperature will lead to faster and more complete hydrolysis, but excessively high temperatures can cause cellulose carbonization or over-hydrolysis.

[0059] Preferably, in step (b03), the filtrate is washed with deionized water until the pH is 6.6–6.9. More preferably, in step (b03), the filtrate is washed with deionized water until the pH is 6.7–6.8. This removes residual hydrochloric acid, preventing acid-catalyzed degradation during subsequent drying or storage, thus improving the chemical stability of microcrystalline cellulose (MCC). When the pH is close to neutral, the acid radicals are essentially removed, which is also required for feed or pharmaceutical excipients.

[0060] Preferably, the drying temperature in step (b04) is 55℃~60℃. More preferably, the drying temperature in step (b04) is 56℃~59℃. Even more preferably, the drying temperature in step (b04) is 57℃~58℃. Removing moisture to ≤5% prevents degradation or yellowing of microcrystalline cellulose (MCC) at high temperatures, ensuring adequate moisture content and maintaining whiteness. The 55℃~60℃ temperature effectively dehydrates the cellulose without causing pyrolysis.

[0061] Preferably, the drying in step (b04) is carried out in an oven. The oven provides a uniform hot air drying environment, can process in batches, accurately control the temperature, ensure uniform moisture content and prevent clumping, and can also be used as an industrial alternative to a fluidized bed dryer.

[0062] Preferably, in step (b05), the particle size is passed through a 180-200 mesh sieve. More preferably, it is passed through a 185-195 mesh sieve. Even more preferably, it is passed through a 190 mesh sieve. Controlling the upper limit of particle size ensures uniform fineness of the finished product, making d 90 ≤120μm, 190 mesh is about 80μm pore size, the material passing through the sieve can meet the particle size requirements.

[0063] Preferably, in step (b05), the d of the microcrystalline cellulose MCC powder 50 50μm~60μm, d 90 ≤120μm, bulk density is 0.28g / cm³ 3 ~0.32g / cm 3 Angle of repose ≤ 33°. Define key quality attributes of microcrystalline cellulose (MCC) to ensure uniformity of subsequent mixing, flowability, and tableting properties. Suitable for stepwise mixing with low-proportion components such as lithocholic acid (LCA). 50The particle size is 50μm to 60μm, which matches the particle size of LCA lithocholic acid premixed powder. The angle of repose is ≤33°, and the flowability is good.

[0064] Preferably, the preparation of the maltodextrin includes the following steps:

[0065] (c01) Prepare corn starch slurry using food-grade corn starch, and adjust the pH of the corn starch slurry to 6.1-6.3 using NaOH solution; provide starch substrate and adjust to the optimal pH of the enzyme to maximize the activity of α-amylase, ensure uniform liquefaction, controllable degree of dextrinization (DE) value, the optimal pH of α-amylase is 6.0-6.5, corn starch is a bulk raw material with the characteristics of low cost and high purity;

[0066] (c02) Add α-amylase to the corn starch slurry from step (c01), stir to liquefy, and then cool.

[0067] (c03) After adjusting the pH of the cooling solution in step (c02) to 4.5-5 using HCl solution, glucosamine is added for saccharification;

[0068] (c04) After removing impurities by pressure filtration of the saccharification solution from step (c03), the filtrate is concentrated to a solid content of 35%–40%, and then spray-dried to obtain a white or light yellow powder of maltodextrin. Corn starch is a bulk food raw material that is stable and readily available; the dual-enzyme liquefaction and saccharification process has good process feasibility; the liquefaction tank, plate and frame filter press, and spray drying tower are all conventional equipment; DE value, moisture, pH, and particle size can all be detected online or offline; the prepared maltodextrin and microcrystalline cellulose (MCC) work synergistically as a carrier for lithocholic acid (LCA), with matching particle sizes.

[0069] Preferably, the dry matter content of the corn starch slurry in step (c01) is 25%–35%. More preferably, the dry matter content of the corn starch slurry is 30%. Determining the balance point between the viscosity and reaction efficiency of the liquefaction system ensures that the starch is fully gelatinized without becoming too viscous and difficult to stir, allowing for sufficient contact between the enzyme and substrate, and a moderate liquefaction time. When the dry matter content is less than 25%, the production capacity is low; when it is greater than 35%, the viscosity is too high, which will cause poor heat and mass transfer.

[0070] Preferably, in step (c01), the concentration of the NaOH solution is 1.0M, and the pH of the corn starch slurry is adjusted to 6.2.

[0071] Preferably, the α-amylase in step (c02) is a thermostable α-amylase.

[0072] Preferably, in step (c02), the α-amylase activity is ≥20000 U / g, and the amount of α-amylase added, based on the dry weight of corn starch, is 0.4‰ to 0.6‰. More preferably, the amount of α-amylase added, based on the dry weight of corn starch, is 0.5‰. Starch is rapidly liquefied at high temperature, hydrolyzing long-chain starch into short-chain dextrins, ensuring the DE value is controlled at 10-15, and the thermoresistant enzyme is stable at 90℃-95℃, endo-hydrolyzing α-1,4 glycosidic bonds.

[0073] Preferably, the liquefaction temperature in step (c02) is 90℃~95℃, and the liquefaction time is 30min~60min. More preferably, the liquefaction temperature is 91℃~94℃, and the liquefaction time is 35min~55min. Even more preferably, the liquefaction temperature is 92℃~93℃, and the liquefaction time is 40min~50min. Controlling the degree of liquefaction to achieve the target degree of dextrinization (DE) is crucial. A moderate dextrin chain length facilitates subsequent saccharification. A slightly higher temperature will increase the liquefaction rate, but excessively high temperatures will cause enzyme inactivation. A longer liquefaction time will result in a higher degree of dextrinization (DE).

[0074] Preferably, the liquefaction endpoint in step (c02) is controlled with a dextrinization degree (DE) value of 10–15. More preferably, the liquefaction endpoint is controlled with a dextrinization degree (DE) value of 11–14. Even more preferably, the liquefaction endpoint is controlled with a dextrinization degree (DE) value of 12–13. A quantitative indicator for the end of liquefaction is defined to ensure batch-to-batch consistency. The final maltodextrin achieves a dextrinization degree (DE) value of 12–14, where DE = reducing sugar / total solids × 100, reflecting the degree of hydrolysis.

[0075] Preferably, in step (CO2), the temperature is cooled to 55°C–60°C. More preferably, it is cooled to 56°C–59°C. Even more preferably, it is cooled to 57°C–58°C. This cooling to the optimal temperature for glucoamylase avoids high-temperature inactivation of glucoamylase, thus maximizing saccharification efficiency. The optimal temperature for glucoamylase is 55–60°C.

[0076] Preferably, in step (CO3), the concentration of the HCl solution is 1.0 M, and the pH of the cooling solution is adjusted to 4.6–4.9. More preferably, the pH of the cooling solution is adjusted to 4.7–4.8. Adjusting to the optimal pH for glucoamylase maximizes the saccharification rate, reduces side reactions, and increases the yield; the optimal pH for glucoamylase is 4.5–5.0.

[0077] Preferably, in step (c03), the glucoamylase activity is ≥100,000 U / g, and the amount of glucoamylase added, based on the dry weight of corn starch, is 0.7‰ to 0.9‰. More preferably, the amount of glucoamylase added, based on the dry weight of corn starch, is 0.8‰. The amount of saccharifying enzyme added is determined to further hydrolyze dextrin into maltose and glucose, increasing the DE value from 10-15 to 12-14, and causing the glucoamylase to exoclease α-1,4 and α-1,6 glycosidic bonds.

[0078] Preferably, the saccharification time in step (c03) is 2 to 3 hours. More preferably, the saccharification time is 2.5 hours. Controlling the degree of saccharification stabilizes the DE value within the target range and avoids excessive glucose production due to over-saccharification. The longer the saccharification time, the higher the DE value, but the increase in DE slows down after 3 hours.

[0079] Preferably, the filtration in step (c04) is performed using a plate and frame filter press. The plate and frame filter press removes insoluble substances such as unhydrolyzed fibers and proteins, improving the purity of the sugar solution. The resulting product has lower ash content and better solubility. Plate and frame filter presses are suitable for solid-liquid separation of materials with high solid content.

[0080] Preferably, in step (c04), the filtrate is concentrated to a solids content of 36%–39%. More preferably, the filtrate is concentrated to a solids content of 37%–38%. Determining the concentration of the feed solution before spray drying balances drying efficiency and atomization effect, resulting in uniform powder particle size and high yield. Too low a solids content will lead to high energy consumption, while too high a solids content will easily clog the nozzle.

[0081] Preferably, the spray drying in step (c04) is performed in a spray drying tower, wherein the atomizer frequency in the spray drying tower is 45Hz to 55Hz. More preferably, the atomizer frequency in the spray drying tower is 50Hz.

[0082] Preferably, in step (c04), the inlet temperature of the spray dryer is 180–190°C, and the outlet temperature is 80–85°C. More preferably, the inlet temperature of the spray dryer is 182–188°C, and the outlet temperature is 81–84°C. Even more preferably, the inlet temperature of the spray dryer is 184–186°C, and the outlet temperature is 82–83°C. The concentrated liquid is converted into dry powder, and the moisture content, particle size, and bulk density are controlled to ensure that the moisture content is ≤5%. 50 Within the required range of 50μm to 60μm, high-temperature instantaneous drying can preserve the low-DE dextrin structure.

[0083] Preferably, the maltodextrin in step (c04) has a moisture content of ≤5.0%, a degree of dextrinization (DE) of 12–14, and an average particle size of d. 50 The particle size is 50 μm to 60 μm. More preferably, the degree of dextrinization (DE) is 13, and the average particle size d...50 The particle size is 55 μm. Key quality properties of maltodextrin were defined to ensure flowability, solubility, and uniformity when premixed with lithocholic acid (LCA), matching the particle size with microcrystalline cellulose (MCC) and lithocholic acid (LCA) for uniform mixing; at a DE of 13, it exhibits moderate water solubility and is non-hygroscopic. 50 At 55 μm, the d with microcrystalline cellulose MCC 50 The values ​​are matched between 50μm and 60μm.

[0084] Preferably, the preparation of the chicken liver powder includes the following steps:

[0085] (d01) Take chicken liver, cut it into pieces and precook it. Then add an equal mass of ice water and crush and homogenize it to obtain a homogenized liquid. Quickly cool down and adjust the material-liquid ratio to prevent the loss of heat-sensitive flavor substances during homogenization. Make the temperature of the homogenized liquid ≤40℃ to retain the volatile flavor. Direct mixing with ice water cools down quickly and dilutes the liquid for easy homogenization.

[0086] (d02) After filtering the homogenized liquid from step (d01), the filtrate is spray-dried to obtain brown chicken liver powder. Raw material availability: Chicken liver is a slaughter byproduct, low in cost and readily available; pre-cooking, homogenization, filtration to spray drying demonstrates good process feasibility; crushers and spray drying towers are standard equipment in food processing; moisture, protein, and fat can all be rapidly detected; the prepared chicken liver powder, as a flavor masking agent, together with lithocholic acid (LCA), microcrystalline cellulose (MCC), and maltodextrin, forms components in the additive, giving it a better flavor.

[0087] Preferably, the chicken liver used in step (d01) is fresh chicken liver selected within 1 to 2 hours after slaughter. Ensuring the freshness of the raw materials reduces fishy smells and spoilage products, preserves natural flavor precursors, and makes the chicken liver powder more palatable and more acceptable to cats. Within 2 hours after slaughter, ATP is not completely degraded, the cell structure is intact, and flavor substances are fully preserved. Pet palatability products have high requirements for the freshness of raw materials.

[0088] Preferably, the chicken liver used in step (d01) is selected from qualified chicken livers after removing fat tissue, blood vessels, and gallbladder. Removing impurities and sources of bitterness improves the purity of the chicken liver powder, reduces off-flavors, and results in a final product with uniform color and pure flavor. Fat is prone to oxidation and rancidity, the gallbladder contains bitter bile acids, and blood vessels contain blood that can affect color.

[0089] Preferably, the chicken liver pieces in step (d01) are 1.5cm in size. 3 ~2.5cm 3 More preferably, the pieces are cut into 2cm cubes. 3 Controlling the uniformity of pre-cooking and crushing ensures that each piece is heated evenly, facilitating subsequent homogenization. This results in the center temperature reaching the standard after pre-cooking, uniform particle size after homogenization, and the 2cm³ block can be fully cooked in the center during 8-10 minutes of pre-cooking.

[0090] Preferably, the pre-cooking temperature in step (d01) is 80℃~85℃, and the pre-cooking time is 8min~10min. More preferably, the pre-cooking temperature is 82℃, and the pre-cooking time is 9min. This denatures, sterilizes, and removes the fishy smell from the protein, fixes the flavor, facilitates subsequent crushing, reduces the fishy smell of chicken liver, and releases the flavor. 80~85℃ coagulates the protein without causing excessive denaturation, otherwise flavor loss will occur. Below 80℃, sterilization will be incomplete, and above 85℃, flavor substances will volatilize.

[0091] Preferably, the crushing and homogenization in step (d01) is completed in a high-speed shear crusher.

[0092] Preferably, the homogenization speed in step (d01) is 4500 rpm to 5000 rpm, and the homogenization time is 1 to 2 minutes. More preferably, the homogenization speed is 4800 rpm, and the homogenization time is 1.5 minutes. This process breaks down chicken liver cells, releasing their contents to form a stable suspension that is easy to spray dry. The resulting suspension has a fine particle size, good solubility after drying, and high-speed shearing disrupts the cell walls and cell membranes, releasing flavor precursors such as proteins, amino acids, and nucleotides.

[0093] Preferably, the filtration in step (d02) is performed through a 180-200 mesh stainless steel screen. More preferably, the filtration is performed through a 185-195 mesh stainless steel screen. Even more preferably, the filtration is performed through a 190 mesh stainless steel screen. This removes unbroken connective tissue and vascular residue, prevents clogging of the spray drying nozzle, and results in a fine finished product with good taste and solubility. The 190 mesh size (approximately 80 μm) effectively filters out insoluble substances such as coarse fibers.

[0094] Preferably, the spray drying in step (d02) is completed in a spray drying tower, wherein the atomizer frequency in the spray drying tower is 45Hz to 55Hz and the atomization pressure is 0.4MPa to 0.6MPa. More preferably, the atomizer frequency in the spray drying tower is 50Hz and the atomization pressure is 0.5MPa. Atomizing the liquid material into fine droplets increases the drying surface area, ensures uniform powder particle size, and prevents sticking to the wall. The higher the frequency and the faster the rotation speed, the slower the droplet velocity; the higher the pressure, the finer the atomization.

[0095] Preferably, in step (d02), the inlet temperature of the spray dryer is 160–170°C, and the outlet temperature is 80–85°C. More preferably, the inlet temperature of the spray dryer is 162–168°C, and the outlet temperature is 81–84°C. Even more preferably, the inlet temperature of the spray dryer is 164–166°C, and the outlet temperature is 82–83°C. This method achieves rapid dehydration, preserves flavor and nutrients, controls moisture content to ≤6%, prevents damage to heat-sensitive substances, ensures that proteins do not undergo excessive denaturation, and maintains good flavor. The inlet temperature of 160–170°C allows for rapid evaporation of moisture while keeping the particle temperature ≤80°C.

[0096] Preferably, the brown chicken liver powder in step (d02) has a moisture content of ≤6.0%, a crude protein content of ≥68%, and a crude fat content of ≤10%. This defines the key quality attributes of the chicken liver powder, ensuring its masking effect, nutritional value, and storage stability. Low moisture prevents clumping, high protein provides flavor, low fat prevents oxidation, microbial growth is prevented when moisture content is <6%, sufficient flavor intensity is achieved when protein content is ≥68%, and good oxidative stability is achieved when fat content is ≤10%.

[0097] Preferably, the preparation of the silica includes the following steps:

[0098] (e01) Dilute industrial water glass with water;

[0099] (e02) Add sulfuric acid solution dropwise to the diluted solution from step (e01) while stirring until precipitation is complete;

[0100] (e03) Continue stirring and aging the solution after complete precipitation in step (e02);

[0101] (e04) Filter the aged solution from step (e03) by pressure filtration, and wash the resulting filter cake with deionized water until the Na+ in the wash solution is removed. + Concentration ≤100ppm;

[0102] (e05) After drying the filter cake from step (e04), pulverize it using an air jet mill and then sieve it to obtain basic silica powder;

[0103] (e06) The basic silica powder from step (e05) is placed in a fluidized bed for preheating, and then the silane coupling agent is sprayed onto the surface of the fluidized powder using nitrogen as a carrier.

[0104] (e07) The material sprayed in step (e06) is subjected to heat treatment to obtain modified silica powder. Industrial water glass is a bulk chemical raw material with good availability; acid precipitation, aging, washing, drying to surface modification have good process feasibility; precipitation reactors, plate and frame filter presses, air jet mills and fluidized beds are all conventional equipment; pH, Na +Both specific surface area and bulk density can be measured, allowing for good quality control; as an anti-caking agent, it ensures that the final additive does not clump during long-term storage, and the quantification is accurate.

[0105] Preferably, the modulus of the industrial water glass in step (e01) is 3.1 to 3.5. More preferably, the modulus of the industrial water glass is 3.2 to 3.4. Even more preferably, the modulus of the industrial water glass is 3.3. By selecting a suitable silicon source and controlling the specific surface area and structure of the precipitated silica, the product can have a high specific surface area and good dispersibility. The modulus is equal to the molar ratio of SiO2 / Na2O. When the modulus is 3.3, the degree of polymerization is moderate and the precipitation is uniform.

[0106] Preferably, in step (e01), the SiO2 content is diluted to 8%–12%. More preferably, it is diluted to 9%–11%. Even more preferably, it is diluted to 10%. By controlling the viscosity and nucleation of the precipitation reaction, uniform silica primary particles are obtained. The larger the specific surface area, the more developed the pore structure. Too low a SiO2 concentration will result in low yield, while too high a concentration will result in severe gelation.

[0107] Preferably, the stirring temperature in step (e02) is 25℃~30℃, and the stirring speed is 250rpm~300rpm. More preferably, the stirring temperature is 26℃~29℃, and the stirring speed is 260rpm~290rpm. Even more preferably, the stirring temperature is 27℃~28℃, and the stirring speed is 270rpm~280rpm. By controlling the precipitation rate and particle growth, fine and uniform silica particles are formed. The narrower the particle size distribution, the more stable the specific surface area. Low temperature and high speed stirring result in more nucleation and finer particles, while high temperature and low speed stirring result in coarser particles.

[0108] Preferably, the addition in step (e02) is a uniform dropwise addition, and the sulfuric acid solution has a mass concentration of 15%. Protons are provided to precipitate silicic acid, controlling the precipitation rate, avoiding localized over-acidity, ensuring a uniform gel, and preventing Na inclusion. + The pH of the system is kept stable by adding the solution at a uniform rate, and the concentration of 15% is appropriate.

[0109] Preferably, the pH of the system is maintained at 6–6.5 during the dropwise addition of step (e02). More preferably, the pH of the system is maintained at 6.1–6.4 during the dropwise addition. Determining the pH at which precipitation is complete ensures complete precipitation of silicic acid while avoiding excessive acidification, resulting in a higher product yield and Na + The residue is low, and silicic acid is completely precipitated when pH < 7; pH 6 to 6.5 is the endpoint for the precipitation of silica.

[0110] Preferably, the aging time in step (e03) is 50 min to 60 min. More preferably, the aging time is 55 min. This allows the precipitated particles to grow and rearrange their structure, improving the stability of the specific surface area, resulting in a well-developed pore structure, improved filtration performance, and the rearrangement of Si-O-Si bonds during the aging process, which can eliminate micro-defects.

[0111] Preferably, the pressure filtration in step (e04) is a plate and frame pressure filtration, and the temperature of the deionized water is 55℃~65℃. More preferably, the temperature of the deionized water is 60℃. Removing the byproduct Na2SO4 reduces conductivity, increases purity, improves modification efficiency, and enhances anti-caking performance. + Residue can affect silane coupling agent grafting, and hot water washing will be more efficient.

[0112] Preferably, the drying temperature in step (e05) is 100℃~105℃, and the drying time is 10h~12h. More preferably, the drying temperature is 102℃, and the drying time is 11h. This removes free water, providing an anhydrous environment for subsequent surface modification. When the moisture content is ≤1%, the silanol groups are adequately retained. The temperature of 100~105℃ can remove physically adsorbed water without damaging the surface silanol groups.

[0113] Preferably, in step (e05), the powder is passed through a 190-210 mesh sieve. More preferably, it is passed through a 195-205 mesh sieve. Even more preferably, it is passed through a 200 mesh sieve. Controlling the upper limit of the basic powder particle size ensures the uniformity of the modification, making d... 90 ≤75μm, 200 mesh is about 75μm, fine powder is beneficial for fluidized bed modification.

[0114] Preferably, in step (e06), the temperature is preheated to 140°C to 150°C. More preferably, it is preheated to 142°C to 148°C. Even more preferably, it is preheated to 145°C. This raises the powder temperature to the modification reaction temperature, improves the grafting efficiency of the silane coupling agent, makes the modified layer uniform, and enhances its hydrophobicity. 140°C to 150°C is the suitable temperature for the condensation reaction between the silane coupling agent and the silanol group.

[0115] Preferably, in step (e06), the mass of the silane coupling agent is 0.5% to 1.5% of the mass of the basic silica powder. More preferably, the mass of the silane coupling agent is 1% of the mass of the basic silica powder. Surface hydrophobic modification reduces the surface energy of silica, prevents agglomeration, results in an angle of repose ≤32°, and excellent flowability. The silane coupling agent reacts with the surface silanol groups to form a monolayer.

[0116] Preferably, the silane coupling agent in step (e06) is a γ-aminopropyltriethoxysilane ethanol solution, and the mass concentration of the γ-aminopropyltriethoxysilane ethanol solution is 8%–12%. More preferably, the mass concentration of the γ-aminopropyltriethoxysilane ethanol solution is 9%–11%. Even more preferably, the mass concentration of the γ-aminopropyltriethoxysilane ethanol solution is 10%. Selecting a suitable coupling agent for silica and introducing aminopropyl groups improves hydrophobicity and ensures that the modified powder can be dispersed in both oil and water systems.

[0117] γ-aminopropyltriethoxysilane KH550 can condense with silanol groups, and the aminopropyl group provides steric hindrance.

[0118] Preferably, the spray rate in step (e06) is 4 mL / min to 6 mL / min. More preferably, the spray rate is 4.5 mL / min to 5.5 mL / min. Even more preferably, the spray rate is 5 mL / min. Controlling the addition rate of the modifier ensures uniform coating, avoids local overwetting, and makes the modified layer uniform and free from agglomeration. Slow spraying is beneficial for monolayer adsorption.

[0119] Preferably, the heat treatment temperature in step (e07) is 140℃~150℃, and the heat treatment time is 20min~40min. More preferably, the heat treatment temperature is 145℃, and the heat treatment time is 30min. This allows the coupling agent to complete the condensation reaction with the silanol group, forming a stable covalent bond, resulting in a long-lasting modification effect and water resistance. 140~150℃ is the optimal temperature for the condensation reaction.

[0120] Preferably, the loose bulk density of the modified silica powder in step (e07) is 0.12 g / cm³ to 0.18 g / cm³, the angle of repose is ≤32°, and the specific surface area is 180 m² / g to 240 m² / g. More preferably, the loose bulk density of the modified silica powder is 0.14 g / cm³ to 0.16 g / cm³, and the specific surface area is 190 m² / g to 230 m² / g. Even more preferably, the loose bulk density of the modified silica powder is 0.15 g / cm³, and the specific surface area is 210 m² / g. The key quality properties of the modified silica are defined to ensure anti-caking, flow-aiding, and hygroscopic properties, enabling uniform dispersion in the premix, preventing LCA agglomeration, resulting in high porosity due to low bulk density, high flowability due to low angle of repose, and high adsorption capacity due to high specific surface area.

[0121] Preferably, the preparation of the tocopherol includes the following steps:

[0122] (f01) Take the deodorized soybean oil distillate, melt it in a water bath, add ethanol and stir to dissolve it; provide a natural source of tocopherol, utilize the by-product of oil refining, turn waste into treasure, the tocopherol content is usually 5-15%, the cost is low, the deodorized soybean oil distillate SODD contains tocopherol, sterol, fatty acid and glycerides, etc.

[0123] (f02) Filter the solution from step (f01), and then perform two-stage molecular distillation on the filtrate to collect the fraction;

[0124] (f03) After concentrating the fraction from step (f02) under reduced pressure, add an equal volume of n-hexane and perform low-temperature crystallization;

[0125] (f04) Filter the substance that has been crystallized in step (f03) to obtain crude tocopherol;

[0126] (f05) The crude tocopherol from step (f04) was subjected to silica gel column chromatography. The target components were collected according to the main peak, and then rotary evaporated to obtain a light yellow oily tocopherol concentrate. Soybean oil deodorized distillate (SODD) is a byproduct of oil refining, widely available and low in cost. The process route from molecular distillation and low-temperature crystallization to column chromatography has good process feasibility. HPLC can quantify various tocopherol homologues, ensuring quality control. As an antioxidant, it protects the oil components in additives, such as the fat and oil phase in chicken liver powder.

[0127] Preferably, the water bath temperature in step (f01) is 50°C to 60°C. More preferably, the water bath temperature is 55°C. This melts the deodorized soybean oil distillate (SODD) and promotes dissolution, reduces viscosity, improves ethanol extraction efficiency, ensures complete dissolution of tocopherols, and minimizes impurities. The 50°C to 60°C melts the fatty acid esters; temperatures above this will compromise heat sensitivity.

[0128] Preferably, in step (f01), the volume of ethanol added is 3 to 5 times the mass of the soybean oil deodorized distillate. More preferably, the volume of ethanol added is 4 times the mass of the soybean oil deodorized distillate. By determining the solvent amount, tocopherols are selectively dissolved, sterols or glycerides are reduced, resulting in a higher extraction rate. The solvent is recoverable, and tocopherols have moderate solubility in 95% ethanol, while sterols have low solubility.

[0129] Preferably, the ethanol in step (f01) is 95% ethanol. This provides a polar solvent to dissolve tocopherols and precipitate some non-polar impurities, thereby increasing the tocopherol content in the initial extract. 95% ethanol has good solubility for tocopherols but poor solubility for fatty acid esters.

[0130] Preferably, the filtration in step (f02) is through a 0.4μm to 0.5μm filter membrane. More preferably, the filtration is through a 0.45μm filter membrane. This removes insoluble matter, protects the molecular distillation equipment, prevents clogging, clarifies the feed, and results in higher distillation efficiency. The 0.45μm membrane can remove particulates and colloids.

[0131] Preferably, in step (f02), the temperature of the first-stage molecular distillation in the two-stage molecular distillation is 100℃~120℃, and the vacuum degree of the first-stage molecular distillation is 8Pa~12Pa; the temperature of the second-stage molecular distillation in the two-stage molecular distillation is 140℃~160℃, and the vacuum degree of the second-stage molecular distillation is 6Pa~10Pa. More preferably, the temperature of the first-stage molecular distillation in the two-stage molecular distillation is 105℃~115℃, and the vacuum degree of the first-stage molecular distillation is 9Pa~11Pa; the temperature of the second-stage molecular distillation in the two-stage molecular distillation is 145℃~155℃, and the vacuum degree of the second-stage molecular distillation is 7Pa~9Pa. Even more preferably, the temperature of the first-stage molecular distillation in the two-stage molecular distillation is 110℃, and the vacuum degree of the first-stage molecular distillation is 10Pa; the temperature of the second-stage molecular distillation in the two-stage molecular distillation is 150℃, and the vacuum degree of the second-stage molecular distillation is 8Pa. The process separates tocopherol from impurities such as fatty acids and sterols. The first stage removes light components such as free fatty acids, while the second stage enriches tocopherol, increasing the tocopherol content from 5-15% to 30-50%. The boiling point of tocopherol is about 200℃ under high vacuum, which is higher than that of fatty acids but lower than that of sterols. The two-stage distillation can achieve continuous separation.

[0132] Preferably, in step (f03), the solution is concentrated to one-third of its original volume under reduced pressure. This increases the solution concentration, creating conditions for low-temperature crystallization, thereby increasing the crystallization yield, reducing the solvent volume, and increasing the supersaturation.

[0133] Preferably, the crystallization temperature in step (f03) is -20°C to -18°C, and the crystallization time is 10h to 12h. More preferably, the crystallization temperature is -19°C, and the crystallization time is 11h. This allows sterols and saturated fatty acid esters to crystallize out, further purifying tocopherols and increasing the tocopherol content to 60-70%. Since sterols have extremely low solubility in hexane at low temperatures, tocopherols will remain in the solution.

[0134] Preferably, in step (f05), the silica gel particle size in the silica gel column is 200-300 mesh, and the column diameter-to-height ratio is 1:13-17. More preferably, the silica gel particle size in the silica gel column is 220-280 mesh, and the column diameter-to-height ratio is 1:14-16. Even more preferably, the silica gel particle size in the silica gel column is 250 mesh, and the column diameter-to-height ratio is 1:15. This process separates tocopherol homologues α, β, γ, and δ, increasing the relative content of δ-tocopherol to achieve a total tocopherol content ≥75% and δ-tocopherol ≥45%. Based on the difference in silica gel adsorption polarity, δ-tocopherol exhibits the highest polarity and is retained most strongly.

[0135] Preferably, the mobile phase in step (f05) of the chromatography process is a mixture of petroleum ether and ethyl acetate, with a volume ratio of petroleum ether to ethyl acetate of 90:10 to 95:5. More preferably, the volume ratio of petroleum ether to ethyl acetate is 91:9 to 94:6. Even more preferably, the volume ratio of petroleum ether to ethyl acetate is 92:8 to 93:7. Elution of tocopherol homologues separates α, β, γ, and δ-tocopherol, resulting in enrichment of δ-tocopherol and improved purity. Based on the polarity differences of δ, γ, β, and α, a higher proportion of ethyl acetate results in stronger elution power.

[0136] Preferably, the tocopherol concentrate in step (f05) contains ≥75% total tocopherol, of which ≥45% is δ-tocopherol. This defines the key quality attributes of the tocopherol concentrate, ensuring its antioxidant effect, inhibiting oil oxidation in additive products, extending shelf life, and highlighting the strongest antioxidant activity of δ-tocopherol. A higher δ-tocopherol content is suitable for industrial antioxidant applications.

[0137] The additive products feature self-designed processes for all six core components, establishing a complete self-sufficiency production system for active ingredients and functional excipients. The supply chain is secure, independent of externally sourced intermediates or excipients. From raw materials to finished products, every step is traceable, ensuring full quality control. Utilizing soybean oil deodorized distillate (SODD) and chicken liver byproducts, or readily available raw materials such as starch, wood pulp, and water glass, costs are minimized. The synergistic design of particle size, density, and flowability ensures uniform mixing and formulation consistency.

[0138] The second technical solution of the present invention: a method for preparing lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following steps: (S01) lithocholic acid, maltodextrin and microcrystalline cellulose are premixed in stages to obtain a basic premix C; a low proportion of lithocholic acid LCA is uniformly dispersed in a large amount of excipients to avoid local enrichment or agglomeration caused by direct mixing, so that the intra-batch uniformity RSD is ≤8% and the dosage accuracy is high. A geometric dilution method of stepwise scale-up mixing is adopted, and each stage is mixed evenly before being mixed with more materials.

[0139] The staged premixing in step (S01) is a three-stage premixing, wherein the three-stage premixing is as follows:

[0140] Lithocholic acid is mixed with 3 to 7 times its mass of maltodextrin to obtain primary premix A; more preferably, lithocholic acid is mixed with 4 to 6 times its mass of maltodextrin; even more preferably, lithocholic acid is mixed with 5 times its mass of maltodextrin; more preferably, the premixing time in the primary premixing process is 5 min to 20 min; even more preferably, the premixing time in the primary premixing process is 10 min to 15 min; the initial dilution ratio is determined to initially disperse lithocholic acid LCA from a high concentration state, reduce lithocholic acid LCA agglomerates, and lay the foundation for subsequent mixing. At a 1:5 ratio, the concentration of lithocholic acid LCA is about 15 to 20%, the viscosity is moderate, and it is easy to mix. Too dilute will cause the subsequent scale-up to be too large, and too thick will cause uneven mixing; ensure that the primary mixing is sufficient to destroy the original lithocholic acid LCA agglomerates, and the initial uniformity of lithocholic acid LCA distribution in primary premix A meets the standard. Too short a mixing time will result in unevenness, and too long a mixing time may cause re-agglomeration or static electricity;

[0141] The primary premix A is mixed with half the mass of the remaining maltodextrin to obtain the secondary premix B; more preferably, the premixing time in the secondary premixing process is 10 min to 30 min; more preferably, the premixing time in the secondary premixing process is 15 min to 25 min; more preferably, the premixing time in the secondary premixing process is 20 min; the initially homogeneous primary premix A is further diluted to reduce the lithocholic acid (LCA) concentration to 5-10%, improving homogeneity and making the LCA distribution in the secondary premix B more uniform. This is the second step in the geometric dilution method, and the mixing difficulty is lower than that of direct mixing from the original powder; ensuring sufficient secondary mixing achieves a uniform distribution of LCA in the secondary premix B, providing a uniform intermediate for the tertiary mixing. The amount of material increases during the secondary mixing, so the mixing time needs to be appropriately extended;

[0142] The secondary premix B, the remaining maltodextrin, and microcrystalline cellulose are mixed together to obtain the basic premix C; more preferably, the premixing time in the basic premixing process is 15 min to 35 min; more preferably, the premixing time in the basic premixing process is 20 min to 30 min; more preferably, the premixing time in the basic premixing process is 25 min; all excipients are added to complete the final mixing, so that the lithocholic acid LCA concentration is diluted to the target level of about 10%, so that the basic premix C reaches the final homogeneity requirement; finally, the microcrystalline cellulose MCC carrier and the remaining maltodextrin are added to complete the geometric dilution; ensuring that the three-stage mixing is sufficient to achieve the final mixing homogeneity, so that the lithocholic acid LCA in the basic premix C is evenly distributed, and all materials are mixed, with the longest total mixing time;

[0143] During the three-stage premixing process, the machine must be stopped after each stage of mixing. Samples should be taken from three horizontal planes within the mixing chamber: the upper third (from the top), the middle (from the middle), and the lower third (from the bottom). At least two points should be taken from the radial center and edge of each horizontal plane, for a total of at least six points. The lithocholic acid content at each point should be detected using HPLC, and the relative standard deviation (RSD) should be calculated. The RSD must be ≤8% before proceeding to the next stage. This process controls mixing uniformity, promptly identifies non-uniformities, and prevents substandard materials from entering the next stage, ensuring that the final product's batch uniformity (RSD ≤8%) is maintained. The sampling points cover three-dimensional space, providing good representativeness.

[0144] (S02) After adding the masking agent to the basic premix C in step (S01) and mixing, tocopherol is added and mixed, and then silica is sprinkled in and mixed to obtain lithocholic acid additive powder. This invention prepares a uniform, stable, and highly fluid lithocholic acid additive powder by premixing lithocholic acid (LCA) with maltodextrin, microcrystalline cellulose (MCC), masking agent, tocopherol, and silica in a three-stage process. This powder is used for subsequent addition to pet food. The step-by-step geometric dilution method is the gold standard for mixing low-dose active ingredients. The three-dimensional motion mixer and HPLC detection demonstrate good process feasibility. Process control at each stage and double inspection of the final product ensure batch consistency. The masking agent coats lithocholic acid (LCA), the tocopherol is oil-dispersed, and the silica is added last, resulting in good matching of material properties. An angle of repose ≤35° ensures that automatic packaging does not clog, and a uniformity RSD ≤8% ensures accurate dosage in small packets.

[0145] Preferably, the staged premixing in step (S01) is performed in a three-dimensional motion mixer. The speed of the three-dimensional motion mixer is 20-30 r / min. After each stage of mixing, multiple samples are taken for HPLC detection, and the RSD is ≤8% before proceeding to the next stage. The three-dimensional motion mixer provides an efficient and gentle mixing method, allowing materials to tumble, shear, and diffuse in three-dimensional space, resulting in high mixing efficiency and no material damage. The mixing uniformity of the three-dimensional motion mixer is superior to that of V-type or double-cone mixers, making it particularly suitable for mixing low-dose active ingredients.

[0146] Preferably, in step (S02), the mixing time after adding the masking agent is 5-10 minutes, the mixing time after adding tocopherol is 5-10 minutes, and the mixing time after sprinkling in silica is 5-15 minutes. More preferably, the mixing time after adding the masking agent is 8 minutes, the mixing time after adding tocopherol is 8 minutes, and the mixing time after sprinkling in silica is 10 minutes. Determining the order of addition and mixing time ensures uniform dispersion of each component. The masking agent coats lithocholic acid (LCA), tocopherol disperses, and silica is added last to aid flow. Adding the masking agent first can cover the unpleasant taste of LCA, the oiliness of tocopherol needs to be fully mixed, and silica is added last to prevent clumping and improve flowability.

[0147] Preferably, multiple samples are taken from the powder premix D in step (S02), and the lithocholic acid content is rapidly determined by HPLC to ensure that the relative standard deviation of the content at each point is ≤8%. The angle of repose of the powder premix D is also measured to ensure that it is ≤35°. Final quality control ensures batch uniformity and flowability, that the product meets expectations, and facilitates subsequent small-packaging and home use. A uniformity RSD ≤8% ensures accurate dosage, and an angle of repose ≤35° provides excellent flowability and prevents clogging of packaging equipment.

[0148] The third technical solution of the present invention: a method for preparing lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following steps: (S01) lithocholic acid, maltodextrin and microcrystalline cellulose are premixed in stages to obtain a basic premix C; a low proportion of lithocholic acid LCA is uniformly dispersed in a large amount of excipients to avoid local enrichment or agglomeration caused by direct mixing, ensuring intra-batch uniformity RSD≤8%, and higher dosage accuracy. A geometric dilution method of stepwise scale-up mixing is adopted, and each stage is mixed evenly before being mixed with more materials.

[0149] The staged premixing in step (S01) is a three-stage premixing, wherein the three-stage premixing is as follows:

[0150] Lithocholic acid is mixed with 3-7 times its mass of maltodextrin to obtain primary premix A; lithochlic acid is then mixed with 4-6 times its mass of maltodextrin; lithochlic acid is further mixed with 5 times its mass of maltodextrin; the premixing time in the primary premixing process is 5-20 min; the initial dilution ratio is determined to initially disperse lithochlic acid LCA from a high concentration state, reduce lithochlic acid LCA agglomerates, and lay the foundation for subsequent mixing. At a 1:5 ratio, the concentration of lithochlic acid LCA is approximately 15-20%, with moderate viscosity, making it easy to mix; ensuring thorough primary mixing breaks down the original lithochlic acid LCA agglomerates, so that the initial uniformity of lithochlic acid LCA distribution in primary premix A initially meets the standard. Too short a mixing time will cause uneven mixing, while too long a time may lead to re-agglomeration or static electricity.

[0151] The primary premix A is mixed with half the mass of the remaining maltodextrin to obtain the secondary premix B. The premixing time in the secondary premixing process is 10 min to 30 min; the premixing time in the secondary premixing process is 15 min to 25 min; the premixing time in the secondary premixing process is 20 min. The initially homogeneous primary premix A is further diluted to reduce the lithocholic acid (LCA) concentration to 5-10% to improve homogeneity, making the LCA distribution of lithocholic acid in the secondary premix B more uniform. This is the second step of the geometric dilution method, and the mixing difficulty is lower than that of direct mixing from the original powder. To ensure sufficient secondary mixing and achieve uniform distribution of LCA in the secondary premix B, a uniform intermediate is provided for the tertiary mixing. The amount of material increases during the secondary mixing, so the mixing time needs to be appropriately extended.

[0152] Secondary premix B, the remaining maltodextrin, and microcrystalline cellulose are mixed together to obtain basic premix C; the premixing time in the basic premixing process is 15 min to 35 min; the premixing time in the basic premixing process is 20 min to 30 min; the premixing time in the basic premixing process is 25 min; all excipients are added to complete the final mixing, diluting the lithocholic acid (LCA) concentration to the target level of about 10%, so that basic premix C reaches the final homogeneity requirement; finally, microcrystalline cellulose (MCC) carrier and the remaining maltodextrin are added to complete geometric dilution; ensuring thorough tertiary mixing to achieve final homogeneity, ensuring uniform distribution of lithocholic acid (LCA) in basic premix C, and mixing all materials, with the longest total mixing time;

[0153] During the three-stage premixing process, the machine must be stopped after each stage of mixing. Samples should be taken from three horizontal planes within the mixing chamber: the upper third (from the top), the middle (from the middle), and the lower third (from the bottom). At least two points should be taken from the radial center and edge of each horizontal plane, for a total of at least six points. The lithocholic acid content at each point should be detected using HPLC, and the relative standard deviation (RSD) should be calculated. The RSD must be ≤8% before proceeding to the next stage. This process controls mixing uniformity, promptly identifies non-uniformities, and prevents substandard materials from entering the next stage, ensuring the final product's uniformity (RSD ≤8%). The sampling points cover three-dimensional space, providing good representativeness.

[0154] (S02) After adding chicken liver powder to the basic premix C in step (S01) and mixing, add tocopherol and mix, then sprinkle in silicon dioxide and mix to obtain lithocholic acid additive powder.

[0155] (S03) Dissolve fish hydrolysate and yeast extract in sunflower seed oil to obtain oil phase solution; provide liquid palatability, enhance cat's willingness to eat, cover the unpleasant flavor of lithocholic acid LCA, and further improve palatability. Suitable for picky cats or wet food. Fish hydrolysate is rich in palatability-enhancing ingredients such as amino acids and nucleotides, yeast extract is rich in umami substances such as nucleotides and glutamic acid, and sunflower seed oil serves as a carrier and enhances aroma. It is a good combination of liquid palatability agents for pet food.

[0156] (S04) The oil phase dissolved in step (S03) is added to the lithocholic acid additive powder in step (S02) by stirring and spraying to obtain lithocholic acid additive liquid; the liquid is evenly distributed on the surface of the powder to form a liquid additive, which can be used directly or as a wet grain ingredient. The liquid product is easier to mix with wet grain or sauce packet. Stirring and spraying can avoid local over-wetting and form a uniform coating. This invention adds a liquid formulation preparation path to the existing powder preparation process for lithocholic acid additives. By dissolving fish hydrolysate and yeast extract in sunflower seed oil and then spraying it onto the powder, a liquid lithocholic acid additive is formed to suit different cats' eating preferences or different diet types, such as wet food or sauce packets. In the powder process, three-stage premixing and process control ensure uniformity and flowability. The addition of a liquid formulation in the liquid process broadens the application scenarios. The powder uses chicken liver powder, while the liquid uses fish hydrolysate and yeast extract, providing dual-path coverage and better palatability. The powder is suitable for dry cat food, while the liquid is suitable for wet cat food or sauce packet-type cat food.

[0157] Preferably, the staged premixing in step (S01) is completed in a three-dimensional motion mixer. The speed of the three-dimensional motion mixer is 20-30 r / min. After each stage of mixing, multiple samples are taken for HPLC detection, and the RSD is ≤8% before proceeding to the next stage. The three-dimensional motion mixer provides an efficient and gentle mixing method, allowing materials to tumble, shear, and diffuse in three-dimensional space, resulting in high mixing efficiency and less material breakage. The mixing uniformity of the three-dimensional motion mixer is superior to that of V-type or double-cone mixers, making it particularly suitable for mixing low-dose active ingredients.

[0158] Preferably, in step (S02), the mixing time after adding chicken liver powder is 5-10 minutes, the mixing time after adding tocopherol is 5-10 minutes, and the mixing time after sprinkling in silica is 5-15 minutes; or the mixing time after adding chicken liver powder is 8 minutes, the mixing time after adding tocopherol is 8 minutes, and the mixing time after sprinkling in silica is 10 minutes. Determining the order of addition and mixing time ensures uniform dispersion of each component. Chicken liver powder coats lithocholic acid (LCA), tocopherol disperses, and silica aids flowability last. Adding chicken liver powder first can mask the unpleasant taste of LCA, the oiliness of tocopherol requires thorough mixing, and adding silica last prevents clumping and improves flowability.

[0159] Preferably, multiple samples are taken from the powder premix D in step (S02), and the lithocholic acid content is rapidly determined by HPLC to ensure that the relative standard deviation of the content at each point is ≤8%. The angle of repose of the powder premix D is also measured to ensure that the angle of repose is ≤35°. Final quality control ensures batch homogeneity and flowability, that the product meets expectations, facilitates subsequent packaging or liquefaction, and that batch homogeneity RSD ≤8% ensures accurate dosage. An angle of repose ≤35° indicates excellent flowability.

[0160] The fourth technical solution of the present invention: the application of lithocholic acid additive for the management of feline obesity or metabolic syndrome, wherein the lithocholic acid additive is mixed into the daily diet of the cat according to the cat's weight, and after continuous feeding for ≥4 weeks, the cat's weight, body condition score, body fat index, abdominal circumference and serum metabolic indicators are detected.

[0161] Continuous feeding resulted in a ≥5% decrease in one of the following from baseline: body weight, BCS score, or abdominal circumference, and / or a ≥10% decrease in triglyceride or total cholesterol levels. The method of use and efficacy assessment cycle for the additive were defined to ensure accurate dosage, high compliance, and quantifiable efficacy. After use, the cat's body weight, BCS score, and abdominal circumference all decreased by ≥5%, and triglycerides (TG) and total cholesterol (TC) all decreased by ≥10%. Mixed feeding did not alter the original diet, making it easy for home administration. Four weeks was the initial metabolic adaptation period for cats, consistent with the typical cycle for pet nutritional interventions. The efficacy endpoint was quantified, providing measurable and comparable indicators for clinical results. It clarified what constitutes effectiveness: a 5% weight loss is clinically significant in cats, corresponding to approximately a 1-point decrease in BCS score; a 10% decrease in blood lipids significantly reduces metabolic risk. The standards were appropriate, both clinically significant and achievable.

[0162] During feeding, serum alanine aminotransferase and aspartate aminotransferase in cats should be tested every 2 to 4 weeks. If either indicator increases by more than 10% from baseline, the dose should be reduced or feeding should be suspended. To prevent the potential hepatotoxicity of lithocholic acid LCA, timely intervention is necessary to avoid cumulative damage. Long-term use is safe and manageable. Lithocholic acid LCA can cause cholestasis at high doses, and a 10% increase is the early warning threshold. This invention incorporates lithocholic acid additives into a cat's daily diet based on body weight. Through continuous feeding for ≥4 weeks, it improves body weight, body fat, and metabolic indicators, and can be used long-term under safety monitoring. The optimal dosage is 30 mg / (kg body weight·day), with a safe adjustment range of 15–45 mg / (kg body weight·day). The efficacy standards are objective, quantifiable, and clinically significant. Safety management employs multi-level early warning systems, from 10%, 30%, to 1.5 times the upper limit, reflecting a cautious approach. The quality control system covers all aspects of content, uniformity, moisture, flowability, and stability. Small, weight-based packaging eliminates the need for weighing, ensuring high compliance and a good user experience. Packaging and storage using aluminum-plastic film, low-humidity dispensing, and room-temperature storage keep costs under control. The normal reference range for adult cat serum ALT is 12–45 U / L; the normal reference range for AST is 10–40 U / L.

[0163] Preferably, the dosage of the lithocholic acid additive is 15–45 mg / (kg body weight·day). More preferably, the dosage of the lithocholic acid additive is 20–40 mg / (kg body weight·day). Even more preferably, the dosage of the lithocholic acid additive is 25–35 mg / (kg body weight·day). Even more preferably, the dosage of the lithocholic acid additive is 30 mg / (kg body weight·day). An effective and safe dosage window is determined to accommodate individual differences such as different body weights and sensitivities. Most cats achieve therapeutic efficacy and tolerate it at 30 mg / kg·day; 30 mg / (kg body weight·day) is the optimal dosage, and 15–45 mg / (kg body weight·day) is a safe adjustment range.

[0164] As a preferred approach, the cat's weight, body condition score, body fat index, abdominal circumference, and serum metabolic indicators should be measured at weeks 4, 8, and every 8 weeks thereafter. The frequency of efficacy assessment should be determined, trends monitored, and the treatment plan adjusted promptly. Initial effects should be observed at week 4, confirmed at week 8, and then maintained every 2 months thereafter. Since cat weight changes are relatively slow, 4 weeks is the minimum meaningful interval.

[0165] Preferably, the serum metabolic indicators include fasting blood glucose, triglycerides, total cholesterol, high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C). A comprehensive assessment of improved glucose and lipid metabolism, monitoring of insulin resistance and dyslipidemia, and multidimensional quantification of improvement in metabolic syndrome are conducted. Obese cats often have elevated fasting blood glucose (GLU), elevated triglycerides (TG), elevated total cholesterol (TC), elevated LDL-C, and low HDL-C.

[0166] Preferably, before starting feeding, at the 2nd week after starting, at the 4th week after starting, and every 4 weeks thereafter, the serum alanine aminotransferase and aspartate aminotransferase levels of the cats are detected. When any one of the serum alanine aminotransferase or aspartate aminotransferase indicators continuously exceeds 1.5 times the upper limit of the detection reference range, or increases by more than 30% compared to the individual's baseline level, the dose of the lithocholic acid additive should be immediately halved or the feeding should be suspended. After the liver function indicators return to normal, feeding can be restarted under monitoring at an addition dose of 15 - 20 mg / (kg body weight·day). More stringent safety management protects individual sensitive cats, maximally avoids liver damage. The 1.5 - fold upper limit is a clear signal of abnormal liver function, and a 30% increase is a significant change, reflecting the emphasis on pet safety.

[0167] Preferably, the relative abundance changes of lithocholic acid derivatives in the cat feces are monitored; the lithocholic acid derivatives include isolithocholic acid, 3 - oxolithocholic acid, 7 - ketolithocholic acid, 12 - ketolithocholic acid, isoallo - lithocholic acid, taurocholate - conjugated lithocholic acid, or glycine - conjugated lithocholic acid. Monitoring the intestinal microbial transformation products, verifying the mechanism hypothesis, evaluating the flora status, and associating with metabolic improvement. Lithocholic acid (LCA) is transformed by intestinal bacteria into various active derivatives, which have immune and metabolic regulatory effects, providing data support for mechanism research.

[0168] Preferably, the HPLC method is used to detect the content of lithocholic acid in the lithocholic acid additive powder. It is qualified within the range of 95% - 105% of the labeled amount of 100 mg / g;

[0169] The RSD of the within - batch uniformity of the lithocholic acid additive powder ≤ 10% is qualified;

[0170] The moisture content in the lithocholic acid additive powder ≤ 5.0% is qualified;

[0171] The angle of repose of the lithocholic acid additive powder ≤ 35° is qualified;

[0172] Three batches of lithocholic acid additive powder samples are taken and placed under accelerated test conditions for 3 months. Samples are taken and detected at the 0th, 1st, 2nd, and 3rd month - ends. The decline rate of the lithocholic acid content ≤ 5.0% is qualified. The product release standards are defined to ensure that each batch of products meets the usage requirements, with the characteristics of accurate dosage, uniform mixing, non - caking, and good fluidity. A 5% content deviation is a conventional requirement, the RSD of batch uniformity ≤ 10% is the standard for low - dose premixes, a moisture content ≤ 5% can prevent mildew, and an angle of repose ≤ 35° ensures smooth packaging and use; the shelf - life stability of the product is evaluated, the changes under normal - temperature storage are predicted, and the shelf - life is inferred to be 12 - 18 months, and a content decline ≤ 5% is an acceptable range.

[0173] Preferably, the accelerated test conditions are a temperature of 38°C to 42°C and a relative humidity of 70% to 80%. More preferably, the accelerated test conditions are a temperature of 40°C and a relative humidity of 75%.

[0174] Preferably, if the lithocholic acid additive powder passes the test, it is then repackaged; during the repackaging process, each package contains a single-day dose or an integer multiple thereof suitable for a cat weighing 5 kg.

[0175] Preferably, each package contains 0.25g to 0.35g during the dispensing process. More preferably, each package contains 0.3g. This method is convenient for home use, eliminating the need for weighing; simply take one package, improving compliance and ensuring accurate dosage. 5kg is a common weight for cats, and 0.3g × 100mg / g = 30mg of lithocholic acid LCA is the optimal dosage, greatly enhancing the user experience.

[0176] Preferably, the dispensing is carried out in a clean room with an ambient humidity of ≤45% using an automatic powder packaging machine and heat-sealed; the packaging bags used for dispensing are made of aluminum-plastic composite film material. This prevents moisture absorption, oxidation, and contamination, ensures product stability during shelf life, prevents moisture increase, prevents LCA lithocholic acid degradation, and the low humidity environment prevents moisture absorption. The aluminum-plastic film also has the characteristics of oxygen barrier, moisture barrier, and light barrier.

[0177] Preferably, the packaged product is placed in a cardboard box and stored in a dry and light-protected environment below 25°C, with a shelf life of 12–18 months. Determining the product storage requirements ensures quality within the product's shelf life, allowing consumers to store it as required. 25°C is considered normal temperature; the environment should be dry and moisture-proof, and light-protected to prevent photodegradation.

[0178] As a preferred option, samples from each batch of products should be retained for at least three full inspections, and stored under the following conditions: 25°C, dry, protected from light, and sealed. This ensures quality traceability, facilitates market sampling and complaint investigations, and the retention quantity meets the requirement of three full inspections, allowing for the reproduction of quality conclusions.

[0179] The present invention has the following beneficial effects:

[0180] (1) Lithocholic acid (LCA) is the main functional component. It can promote the secretion of GLP-1 by activating the intestinal TGR5 receptor, thereby improving energy metabolism and blood glucose and blood lipid levels, and thus regulating feline obesity and metabolic syndrome. Maltodextrin is the main diluent. It can adjust the content of lithocholic acid to a suitable dietary dosage range and ensure the accuracy of measurement during packaging. It also has a certain taste-masking effect, which is beneficial for long-term feeding. Microcrystalline cellulose (MCC) is a safe and inert carrier. It has good adsorption and dilution effects, which can ensure the uniform distribution of lithocholic acid in the additive and improve the overall fluidity and processing performance. The taste masker has a significant flavor-improving effect. It can mask the bitterness and odor of lithocholic acid, improve the cat's feeding compliance, and ensure the sustainability of the intervention effect. Silica is an adjuvant. It plays an anti-caking role, which can reduce the agglomeration of powder during storage and packaging, maintain the fluidity and stability of the premix, and facilitate industrial production. Tocopherol is an adjuvant. It plays an antioxidant role, which can effectively inhibit the oxidative degradation of lithocholic acid and oil components in the formula, improve the stability of the formulation, and extend the shelf life of the product.

[0181] (2) Provide a basic formulation framework to ensure that lithocholic acid LCA is uniformly dispersed, has good flowability, acceptable palatability and antioxidant stability. A limited number of microcrystalline cellulose and maltodextrin serve as an inert skeleton to carry and dilute lithocholic acid LCA, which is convenient for subsequent cat weight measurement. A limited number of masking agents cover the bitterness and irritation of lithocholic acid LCA. The stepwise mixing achieves uniform distribution of low-content active ingredients, and finally prepares an additive with controllable content and intra-batch uniformity RSD≤10%, which supports selective and precise feeding at 15~45mg / (kg body weight·day) according to the specific tolerance of the cat.

[0182] (3) When used in prescription diets, it can be added to the diet according to the actual situation. It does not require changing the prescription diet and is convenient for home use. It can effectively reduce the weight, abdominal circumference, body condition score (BCS) and body fat index (FBMI) of obese cats. It can improve the blood sugar and blood lipids of obese cats, thereby reducing the levels of triglycerides (TG), total cholesterol (TC), and low-density lipoprotein cholesterol (LDL). It can enhance the antioxidant capacity, thereby increasing the level of glutathione peroxidase (GSH-Px) and decreasing the level of malondialdehyde (MDA) in obese cats. It can also enhance the humoral immunity of obese cats, thereby increasing the levels of immunoglobulin AIg, immunoglobulin GIgG, and immunoglobulin MIgM. It can also improve the low-grade inflammation of obese cats, thereby reducing the levels of pro-inflammatory cytokines interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α), and increasing the level of anti-inflammatory cytokines interleukin-10 (IL-10). It has the characteristics of effectively improving the weight of obese cats, effectively improving glucose and blood lipid metabolism, and providing multiple health benefits.

[0183] (4) Lithocholic acid (LCA) is prepared into a uniform, stable and free-flowing lithocholic acid additive powder by three-stage premixing with maltodextrin, microcrystalline cellulose (MCC), flavor masking agent, tocopherol and silica. This powder is then used for subsequent addition to pet food. The stepwise geometric dilution method is the gold standard for mixing low-dose active ingredients. The three-dimensional motion mixer and HPLC detection both have good process feasibility. Each stage of process control and the final product double inspection ensure batch consistency. The flavor masking agent coats LCA, the tocopherol is oily and dispersed, and silica is added last, resulting in good matching of the properties between the materials. An angle of repose ≤35° ensures that the automatic packaging will not be blocked, and a uniformity RSD ≤8% ensures accurate dosage in small bags.

[0184] (5) Based on the original powder preparation process of lithocholic acid additive, a liquid preparation path has been added. By dissolving fish hydrolysate and yeast extract in sunflower seed oil and then spraying it onto the powder, liquid lithocholic acid additive is formed to adapt to different cats' eating preferences or different types of daily food, such as wet food or sauce packets. In the powder process, three-stage premixing and process control ensure uniformity and flowability. In the liquid process, the application scenarios have been broadened by adding liquid dosage forms. The powder is designed to use chicken liver powder, and the liquid uses fish hydrolysate and yeast extract, with dual-path coverage and better palatability. The powder is suitable for dry cat food, and the liquid is suitable for wet cat food or sauce packet cat food.

[0185] (6) Lithocholic acid additives are mixed into the daily diet of cats by weight measurement. Through continuous feeding for ≥4 weeks, the weight, body fat and metabolic indicators are improved and long-term use is carried out under the premise of safety monitoring. The optimal dose is 30mg / (kg body weight·day), and the safe adjustment range is 15~45mg / (kg body weight·day). The efficacy standard is objective, quantifiable and clinically significant. In terms of safety management, multi-level early warning is adopted, from 10%, 30% to 1.5 times the upper limit, which reflects a cautious attitude. The quality control system includes full coverage of content, uniformity, moisture, flowability and stability. Small bags are packaged according to weight, eliminating the need for weighing, resulting in high compliance and a good user experience. The packaging and storage of aluminum-plastic film, low humidity packaging and room temperature storage are cost-controllable. Attached Figure Description

[0186] Figure 1This graph shows the changes in bile acid profiles in the feces of the control (CON) and obese OB cats in the experimental case, comparing the levels of lithocholic acid and its intestinal microbial derivatives: lithocholic acid, 7-ketolithocholic acid, 12-ketolithocholic acid, isolithocholic acid, taurolithocholic acid, dehydrolithocholic acid, gycolithocholic acid, and isoallolithocholic acid. If the graph is marked with a significance symbol, such as *, **, ***, or ****, it indicates that the difference compared to baseline is statistically significant (p < 0.05, p < 0.01, p < 0.001, or p < 0.0001, respectively).

[0187] Figure 2 The figure shows the comparison of body weight between the control group (CON cats) and the obese group (OB cats) in the experimental case, as well as the effect of adding 30 mg / kg / d of lithocholic acid to the diet of obese cats on their body weight, average daily weight gain and food intake.

[0188] Figure 3 The baseline differences in chest circumference, abdominal circumference, body condition score (BCS), body fat index (FBMI), and serum metabolic indicators (fasting blood glucose (GLU), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and triglycerides (TG) between the control group (CON) and the obese group (OB) in the experimental case, as well as the changes in the above body dimensions and metabolic indicators in the obese group after 6 weeks of intervention with lithocholic acid supplemented at 30 mg / kg / day according to body weight; BCS was assessed using the WSAVA nine-point scale, and FBMI was calculated based on chest circumference and hind limb length; if a significance symbol such as *, **, ***, or **** is marked in the figure, it represents a statistically significant difference compared to baseline, p < 0.05, p < 0.01, p < 0.001, or p < 0.0001, respectively;

[0189] Figure 4 The figures show the changes in serum antioxidant, immune, and inflammatory factors in cats after intervention with lithocholic acid in the experimental case. These include total antioxidant capacity (T-AOC), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA); immunoglobulins (AIgA, GIgG, and MIgM); pro-inflammatory cytokines (interleukin-6 IL-6, interleukin-1β IL-1β, tumor necrosis factor-α TNF-α); and anti-inflammatory cytokines (interleukin-10 IL-10). Significance symbols such as *, **, ***, or **** indicate statistically significant differences compared to baseline, p < 0.05, p < 0.01, p < 0.001, or p < 0.0001, respectively.

[0190] Figure 5 The figure shows the effect of lithocholic acid additive intervention on the levels of ALB, ALT, AST, Glob, A, and G in cat serum in the experimental case. Detailed Implementation

[0191] The present invention will be further described below with reference to the accompanying drawings and embodiments, but this should not be construed as limiting the present invention.

[0192] Lithocholic acid additives for the management of obesity or metabolic syndrome in cats comprise the following components by weight: 5-15 parts lithocholic acid, 70-88 parts maltodextrin, 5-20 parts microcrystalline cellulose, 1-5 parts flavor masking agent, 0.2-1.0 parts silica, and 0.05-0.2 parts tocopherol.

[0193] Lithocholic acid additives for the management of obesity or metabolic syndrome in cats comprise the following components by weight: 8-12 parts lithocholic acid, 75-85 parts maltodextrin, 8-18 parts microcrystalline cellulose, 2-4 parts flavor masking agent, 0.4-0.8 parts silica, and 0.08-0.18 parts tocopherol.

[0194] Lithocholic acid additives for the management of obesity or metabolic syndrome in cats comprise the following components by weight: 9-11 parts lithocholic acid, 78-82 parts maltodextrin, 10-15 parts microcrystalline cellulose, 2-4 parts flavor masking agent, 0.5-0.7 parts silica, and 0.1-0.15 parts tocopherol.

[0195] Lithocholic acid additive for the management of obesity or metabolic syndrome in cats comprises the following components by weight: 10 parts lithocholic acid, 80 parts maltodextrin, 12 parts microcrystalline cellulose, 3 parts flavor masking agent, 0.6 parts silica, and 0.12 parts tocopherol.

[0196] Lithocholic acid additives for the management of obesity or metabolic syndrome in cats also include the following components by weight: 90-100 parts sunflower seed oil, 1-2 parts fish hydrolysate, and 1-2 parts yeast extract.

[0197] Lithocholic acid additives for the management of obesity or metabolic syndrome in cats also include the following components by weight: 92-98 parts sunflower seed oil, 1.2-1.8 parts fish hydrolysate, and 1.2-1.8 parts yeast extract.

[0198] The lithocholic acid additive for the management of obesity or metabolic syndrome in cats also includes the following components by weight: 95 parts sunflower seed oil, 1.5 parts fish hydrolysate, and 1.5 parts yeast extract.

[0199] The masking agent is selected from at least one of chicken liver powder, fish hydrolysate, or yeast extract.

[0200] Lithocholic acid is obtained by enzymatically directed dehydroxylation of ursodeoxycholic acid as a starting substrate. The enzymatically directed dehydroxylation reaction of lithochlic acid includes the following steps:

[0201] (a01) Enzyme system preparation

[0202] (a01-1) Inoculate Clostridium hiranonis strain into enhanced Clostridium medium for anaerobic culture; Clostridium hiranonis strain preservation number is JCM16549; the culture temperature in step (a01-1) is 35℃~38℃, and the culture time is 36h~60h; the culture temperature is 36℃~37℃, and the culture time is 42h~54h; the culture temperature is 37℃, and the culture time is 48h.

[0203] (a01-2) Collect the bacterial cells by centrifugation of the culture medium from step (a01-1), then sonicate and centrifuge to obtain the supernatant, and obtain crude enzyme solution;

[0204] (a01-3) Add ammonium sulfate to the crude enzyme solution in step (a01-2) for fractional precipitation, and collect the precipitate with a saturation of 30% to 60%;

[0205] (a01-4) Dissolve the precipitate in step (a01-3) with sterile water, then purify it by DEAE-Sepharose FastFlow ion exchange chromatography, collect the fraction with 12α-hydroxyl removal activity, freeze dry it to obtain enzyme powder; in step (a01-4), the specific activity of the enzyme powder is not less than 15 U / mg protein.

[0206] (a02) Enzymatic reaction

[0207] (a02-1) Dissolve ursodeoxycholic acid (UDCA) with a purity ≥98% in a mixed solution of phosphate buffer and methanol to obtain a substrate solution; the concentration of the substrate solution in step (a02-1) is 50 mg / mL; the pH of the mixed solution in step (a02-1) is 6.6–6.9, and the volume ratio of phosphate buffer to methanol in the mixed solution is 3.5–4.5:1; the pH of the mixed solution in step (a02-1) is 6.7–6.8, and the volume ratio of phosphate buffer to methanol in the mixed solution is 3.8–4.2:1.

[0208] (a02-2) Add enzyme powder to the substrate solution in step (a02-1) and stir to react; in step (a02-2), the mass of enzyme powder added is 6%–10% of the mass of the substrate solution; the mass of enzyme powder added is 7%–9% of the mass of the substrate solution; the mass of enzyme powder added is 8% of the mass of the substrate solution; the reaction temperature in step (a02-2) is 36℃–37℃, the reaction time is 28h–30h, and the pH of the reaction system is 6.7–6.9; the reaction temperature in step (a02-2) is 36.5℃, the reaction time is 29h, and the pH of the reaction system is 6.8; the reaction in step (a02-2) is carried out in a constant temperature oscillating reactor;

[0209] (a03) Product purification

[0210] (a03-1) After the reaction in step (a02-2), add an equal volume of ethyl acetate to the mixture and perform multiple extractions. Combine the organic phases and evaporate to dryness. The number of extractions in step (a03-1) is 2 to 5, and the evaporation is performed by rotary evaporation. The number of extractions is 3 to 4.

[0211] (a03-2) The evaporated product from step (a03-1) is dissolved in hot ethanol and hot filtered, and then the filtrate is allowed to stand for aging; the volume of hot ethanol in step (a03-2) is 9 to 11 times the volume of the evaporated product; the volume of hot ethanol is 10 times the volume of the evaporated product; the aging temperature in step (a03-2) is 3℃ to 6℃, and the aging time is 6h to 18h; the aging temperature in step (a03-2) is 4℃ to 5℃, and the aging time is 9h to 15h; the aging temperature in step (a03-2) is 4.5℃, and the aging time is 12h; the hot ethanol in step (a03-2) is 95% hot ethanol;

[0212] (a03-3) The white crystals aged in step (a03-2) are filtered and washed, and then vacuum dried to obtain lithocholic acid (LCA); the vacuum drying temperature in step (a03-3) is 35℃~45℃, the vacuum drying pressure is -0.1MPa~-0.09MPa, and the vacuum drying time is 3h~9h; the vacuum drying temperature in step (a03-3) is 38℃~42℃, the vacuum drying pressure is -0.095MPa, and the vacuum drying time is 4h~8h; the vacuum drying temperature in step (a03-3) is 40℃, the vacuum drying pressure is -0.095MPa, and the vacuum drying time is 6h; the purity of lithocholic acid (LCA) in step (a03-3) is ≥95%.

[0213] The preparation of microcrystalline cellulose (MCC) includes the following steps:

[0214] (b01) Take bleached wood pulp with α-cellulose content ≥90%, dry it, and then pulverize it; in step (b01), the pulp is pulverized to a fiber length of 1mm to 2mm;

[0215] (b02) After adding hydrochloric acid solution to the pulverized material in step (b01) and stirring, the mixture is rapidly cooled to room temperature with ice water; the solid-liquid ratio of the pulverized material to the hydrochloric acid solution in step (b02) is 1:10 to 20; the solid-liquid ratio of the pulverized material to the hydrochloric acid solution is 1:12 to 18; the solid-liquid ratio of the pulverized material to the hydrochloric acid solution is 1:15; the concentration of the hydrochloric acid solution in step (b02) is 2N; the reaction temperature in step (b02) is 80℃ to 85℃, and the reaction time is 80 min to 120 min; the reaction temperature is 81℃ to 84℃, and the reaction time is 90 min to 110 min; the reaction temperature is 82℃ to 83℃, and the reaction time is 95 min to 105 min;

[0216] (b03) After filtering the cooling mixture from step (b02), take the filtrate and wash it with deionized water until the pH of the filtrate is 6.5-7; in step (b03), wash it with deionized water until the pH of the filtrate is 6.6-6.9; in step (b03), wash it with deionized water until the pH of the filtrate is 6.7-6.8.

[0217] (b04) Dry the filtrate after washing in step (b03) until the moisture content is ≤5%; the drying temperature in step (b04) is 55℃~60℃; the drying temperature in step (b04) is 56℃~59℃; the drying temperature in step (b04) is 57℃~58℃; the drying in step (b04) is carried out in an oven.

[0218] (b05) The dried material from step (b04) is pulverized by an air jet mill and then sieved to obtain microcrystalline cellulose (MCC) powder; in step (b05), it is sieved through a 180-200 mesh sieve; through a 185-195 mesh sieve; through a 190 mesh sieve; the d of the microcrystalline cellulose (MCC) powder in step (b05) 50 50μm~60μm, d 90 ≤120μm, bulk density is 0.28g / cm³ 3 ~0.32g / cm 3 The angle of repose is ≤33°.

[0219] The preparation of maltodextrin includes the following steps:

[0220] (c01) Prepare corn starch slurry using food-grade corn starch, and adjust the pH of the corn starch slurry to 6.1-6.3 using NaOH solution; the dry matter content of the corn starch slurry in step (c01) is 25%-35%; the dry matter content of the corn starch slurry is 30%; the concentration of NaOH solution in step (c01) is 1.0M, and the pH of the corn starch slurry is adjusted to 6.2;

[0221] (c02) α-Amylase is added to the corn starch slurry from step (c01), stirred, liquefied, and then cooled; the α-amylase in step (c02) is a thermoresistant α-amylase; the enzyme activity of the α-amylase in step (c02) is ≥20000 U / g, and the amount of α-amylase added, based on the dry weight of corn starch, is 0.4‰~0.6‰; the amount of α-amylase added, based on the dry weight of corn starch, is 0.5‰; the liquefaction temperature in step (c02) is 90℃~95℃, and the liquefaction time is 30min~60min. n; liquefaction temperature is 91℃~94℃, liquefaction time is 35min~55min; liquefaction temperature is 92℃~93℃, liquefaction time is 40min~50min; the liquefaction endpoint in step (c02) is controlled at a dextrinization degree DE value of 10~15; the liquefaction endpoint is controlled at a dextrinization degree DE value of 11~14; the liquefaction endpoint is controlled at a dextrinization degree DE value of 12~13; in step (c02), cooling is performed to 55℃~60℃; cooling is performed to 56℃~59℃; cooling is performed to 57℃~58℃;

[0222] (c03) After adjusting the pH of the cooling solution in step (c02) to 4.5–5 using HCl solution, glucoamylase is added for saccharification; the concentration of the HCl solution in step (c03) is 1.0M, and the pH of the cooling solution is adjusted to 4.6–4.9; the pH of the cooling solution is adjusted to 4.7–4.8; the enzyme activity of glucoamylase in step (c03) is ≥100,000 U / g, and the amount of glucoamylase added is 0.7‰–0.9‰ based on the dry weight of corn starch; the amount of glucoamylase added is 0.8‰ based on the dry weight of corn starch; the saccharification time in step (c03) is 2h–3h; the saccharification time is 2.5h.

[0223] (c04) After removing impurities by pressure filtration of the saccharified solution in step (c03), the filtrate is concentrated to a solid content of 35%–40%, and then spray-dried to obtain a white or light yellow powder of maltodextrin; the pressure filtration in step (c04) is completed using a plate and frame filter press; the filtrate in step (c04) is concentrated to a solid content of 36%–39%; the filtrate is further concentrated to a solid content of 37%–38%; the spray drying in step (c04) is completed in a spray drying tower, and the atomizer in the spray drying tower is frequently... The frequency is 45Hz~55Hz; the atomizer frequency in the spray drying tower is 50Hz; the inlet temperature of spray drying in step (c04) is 180~190℃, and the outlet temperature is 80~85℃; the inlet temperature of spray drying is 182~188℃, and the outlet temperature is 81~84℃; the inlet temperature of spray drying is 184~186℃, and the outlet temperature is 82~83℃; the moisture content of maltodextrin in step (c04) is ≤5.0%, the degree of dextrinization (DE) is 12~14, and the average particle size is d 50 The particle size is 50μm to 60μm; the degree of dextrinization (DE) is 13, and the average particle size is d. 50 It is 55μm.

[0224] The preparation of chicken liver powder includes the following steps:

[0225] (d01) Cut chicken liver into pieces and pre-cook. Then add an equal amount of ice water and homogenize to obtain a homogenized liquid. The chicken liver used in step (d01) should be fresh chicken liver selected within 1-2 hours after slaughter. The chicken liver used in step (d01) should be chicken liver that has passed inspection and has had its fat tissue, blood vessels, and gallbladder removed. The chicken liver pieces in step (d01) should be 1.5cm in size. 3 ~2.5cm 3 Cut into 2cm pieces 3 The pre-cooking temperature in step (d01) is 80℃~85℃, and the pre-cooking time is 8min~10min; the pre-cooking temperature is 82℃, and the pre-cooking time is 9min; the crushing and homogenization in step (d01) is completed in a high-speed shear crusher; the homogenization speed in step (d01) is 4500rpm~5000rpm, and the homogenization time is 1~2min; the homogenization speed is 4800rpm, and the homogenization time is 1.5min;

[0226] (d02) After filtering the homogenized liquid from step (d01), the filtrate is spray-dried to obtain brown chicken liver powder; the filtration in step (d02) is completed through a 180-200 mesh stainless steel sieve; the filtration is completed through a 185-195 mesh stainless steel sieve; the filtration is completed through a 190 mesh stainless steel sieve; the spray drying in step (d02) is completed in a spray drying tower, the atomizer frequency in the spray drying tower is 45Hz-55Hz, and the atomization pressure is 0.4MPa-0.6MPa; The atomizer frequency in the spray drying tower is 50Hz, and the atomization pressure is 0.5MPa; in step (d02), the inlet temperature of the spray drying is 160-170℃, and the outlet temperature is 80-85℃; the inlet temperature of the spray drying is 162-168℃, and the outlet temperature is 81-84℃; the inlet temperature of the spray drying is 164-166℃, and the outlet temperature is 82-83℃; the moisture content of the brown chicken liver powder in step (d02) is ≤6.0%, the crude protein content is ≥68%, and the crude fat content is ≤10%.

[0227] The preparation of silica includes the following steps:

[0228] (e01) Dilute industrial water glass with water; the modulus of the industrial water glass in step (e01) is 3.1-3.5; the modulus of the industrial water glass is 3.2-3.4; the modulus of the industrial water glass is 3.3; in step (e01), dilute to a SiO2 content of 8%-12%; dilute to a SiO2 content of 9%-11%; dilute to a SiO2 content of 10%;

[0229] (e02) Add sulfuric acid solution dropwise to the diluted solution in step (e01) under stirring until precipitation is complete; the stirring temperature in step (e02) is 25℃~30℃, and the stirring speed is 250rpm~300rpm; the stirring temperature is 26℃~29℃, and the stirring speed is 260rpm~290rpm; the stirring temperature is 27℃~28℃, and the stirring speed is 270rpm~280rpm; the dropwise addition in step (e02) is uniform, and the mass concentration of the sulfuric acid solution is 15%; the pH of the system is maintained at 6~6.5 during the dropwise addition in step (e02); the pH of the system is maintained at 6.1~6.4 during the dropwise addition.

[0230] (e03) Continue stirring and aging the solution after complete precipitation in step (e02); the aging time in step (e03) is 50 min to 60 min; the aging time is 55 min.

[0231] (e04) Filter the aged solution from step (e03) by pressure filtration, and wash the resulting filter cake with deionized water until the Na+ in the wash solution is removed. +Concentration ≤ 100 ppm; the filter press in step (e04) is a plate and frame filter press, and the temperature of the deionized water is 55℃~65℃; the temperature of the deionized water is 60℃;

[0232] (e05) After drying the filter cake from step (e04), pulverize it using an air jet mill and then sieve it to obtain basic silica powder; the drying temperature in step (e05) is 100℃~105℃, and the drying time is 10h~12h; the drying temperature is 102℃, and the drying time is 11h; in step (e05), the powder is sieved through a 190-210 mesh sieve; through a 195-205 mesh sieve; and through a 200 mesh sieve;

[0233] (e06) The basic silica powder from step (e05) is placed in a fluidized bed for preheating, and then a silane coupling agent is sprayed onto the surface of the fluidized powder using nitrogen as a carrier; in step (e06), the preheating temperature is 140℃~150℃; preheating temperature is 142℃~148℃; preheating temperature is 145℃; in step (e06), the mass of the silane coupling agent is 0.5%~1.5% of the mass of the basic silica powder; the mass of the silane coupling agent is 1% of the mass of the basic silica powder; in step (e06), the silane... The coupling agent is a γ-aminopropyltriethoxysilane ethanol solution with a mass concentration of 8%–12%; a mass concentration of γ-aminopropyltriethoxysilane ethanol solution of 9%–11%; a mass concentration of γ-aminopropyltriethoxysilane ethanol solution of 10%; the spray rate in step (e06) is 4 mL / min–6 mL / min; the spray rate is 4.5 mL / min–5.5 mL / min; the spray rate is 5 mL / min.

[0234] (e07) The material sprayed in step (e06) is subjected to heat treatment to obtain modified silica powder; the heat treatment temperature in step (e07) is 140℃~150℃ and the heat treatment time is 20min~40min; the heat treatment temperature is 145℃ and the heat treatment time is 30min; the loose density of the modified silica powder in step (e07) is 0.12g / cm³~0.18g / cm³, the angle of repose is ≤32°, and the specific surface area is 180m² / g~240m² / g; the loose density of the modified silica powder is 0.14g / cm³~0.16g / cm³ and the specific surface area is 190m² / g~230m² / g; the loose density of the modified silica powder is 0.15g / cm³ and the specific surface area is 210m² / g.

[0235] The preparation of tocopherol includes the following steps:

[0236] (f01) Melt the deodorized soybean oil distillate in a water bath, then add ethanol and stir to dissolve; the water bath temperature in step (f01) is 50℃~60℃; the water bath temperature is 55℃; the volume of ethanol added in step (f01) is 3~5 times the mass of the deodorized soybean oil distillate; the volume of ethanol added is 4 times the mass of the deodorized soybean oil distillate; the ethanol in step (f01) is 95% ethanol;

[0237] (f02) Filter the solution from step (f01), then perform two-stage molecular distillation on the filtrate and collect the fractions; the filtration in step (f02) is through a 0.4μm to 0.5μm filter membrane; the filtration in step (f02) is through a 0.45μm filter membrane; the temperature of the first stage of molecular distillation in step (f02) is 100℃ to 120℃, and the vacuum degree of the first stage of molecular distillation is 8Pa to 12Pa; the temperature of the second stage of molecular distillation in step (f02) is 140℃ to 160℃, and the vacuum degree of the second stage of molecular distillation is 6Pa to 12Pa. 10 Pa; In two-stage molecular distillation, the temperature of the first stage is 105℃~115℃, and the vacuum degree of the first stage is 9Pa~11Pa; In two-stage molecular distillation, the temperature of the second stage is 145℃~155℃, and the vacuum degree of the second stage is 7Pa~9Pa; In two-stage molecular distillation, the temperature of the first stage is 110℃, and the vacuum degree of the first stage is 10Pa; In two-stage molecular distillation, the temperature of the second stage is 150℃, and the vacuum degree of the second stage is 8Pa.

[0238] (f03) After concentrating the fraction in step (f02) under reduced pressure, add an equal volume of n-hexane and crystallize at low temperature; in step (f03), concentrate under reduced pressure to 1 / 3 of the original volume; the crystallization temperature in step (f03) is -20℃ to -18℃, and the crystallization time is 10h to 12h; the crystallization temperature is -19℃, and the crystallization time is 11h.

[0239] (f04) Filter the substance that has been crystallized in step (f03) to obtain crude tocopherol;

[0240] (f05) The crude tocopherol from step (f04) is subjected to silica gel column chromatography. The target component is collected according to the main peak and then rotary evaporated to obtain a light yellow oily tocopherol concentrate. The silica gel particles in the silica gel column in step (f05) are 200-300 mesh, and the column diameter-to-height ratio is 1:13-17. The silica gel particles in the silica gel column are 220-280 mesh, and the column diameter-to-height ratio is 1:14-16. The silica gel particles in the silica gel column are 250 mesh, and the column diameter-to-height ratio is 1:15. The mobile phase in the chromatography process of step (f05) is a mixture of petroleum ether and ethyl acetate, with a volume ratio of petroleum ether to ethyl acetate of 90:10-95:5, 91:9-94:6, and 92:8-93:7. The total tocopherol content in the tocopherol concentrate of step (f05) is ≥75%, of which δ-tocopherol is ≥45%.

[0241] A method for preparing a lithocholic acid additive for the management of obesity or metabolic syndrome in cats includes the following steps:

[0242] (S01) Lithocholic acid, maltodextrin and microcrystalline cellulose are fractionally premixed to obtain basic premix C; the fractional premixing in step (S01) is completed in a three-dimensional motion mixer;

[0243] The staged premixing in step (S01) is a three-stage premixing, and the three-stage premixing is as follows:

[0244] Lithocholic acid is mixed with 3 to 7 times its mass of maltodextrin to obtain primary premix A; lithochlic acid is mixed with 4 to 6 times its mass of maltodextrin; lithochlic acid is mixed with 5 times its mass of maltodextrin; the premixing time in the primary premixing process is 5 min to 20 min; the premixing time in the primary premixing process is 10 min to 15 min.

[0245] Mix the primary premix A with half the mass of the remaining maltodextrin to obtain the secondary premix B; the premixing time in the secondary premixing process is 10 min to 30 min; the premixing time in the secondary premixing process is 15 min to 25 min; the premixing time in the secondary premixing process is 20 min.

[0246] The secondary premix B, the remaining maltodextrin, and microcrystalline cellulose are mixed together to obtain the basic premix C; the premixing time in the basic premixing process is 15 min to 35 min; the premixing time in the basic premixing process is 20 min to 30 min; the premixing time in the basic premixing process is 25 min.

[0247] During the three-stage premixing process, the machine must be stopped after each stage of mixing is completed. At least two points should be taken from the upper part (1 / 3 of the distance from the top), the middle part (1 / 2 of the distance from the top), and the lower part (1 / 3 of the distance from the bottom) of the mixing chamber, with a total of at least six points. The lithocholic acid content at each point should be detected by HPLC, and the relative standard deviation (RSD) should be calculated. Only when the RSD is ≤8% can the next step be carried out.

[0248] (S02) After adding a masking agent to the basic premix C in step (S01) and mixing, add tocopherol and mix, then sprinkle in silica and mix to obtain lithocholic acid additive powder; the mixing time after adding the masking agent in step (S02) is 5 min to 10 min, the mixing time after adding tocopherol is 5 min to 10 min, and the mixing time after sprinkling in silica is 5 min to 15 min; the mixing time after adding the masking agent is 8 min, the mixing time after adding tocopherol is 8 min, and the mixing time after sprinkling in silica is 10 min; multiple samples are taken from the powder premix D in step (S02), and the lithocholic acid content is quickly detected by HPLC to ensure that the relative standard deviation of the content at each point is ≤8%, and the angle of repose of the powder premix D is measured to ensure that the angle of repose is ≤35°.

[0249] A method for preparing a lithocholic acid additive for the management of obesity or metabolic syndrome in cats includes the following steps:

[0250] (S01) Lithocholic acid, maltodextrin and microcrystalline cellulose are fractionally premixed to obtain basic premix C; the fractional premixing in step (S01) is completed in a three-dimensional motion mixer;

[0251] The staged premixing in step (S01) is a three-stage premixing, and the three-stage premixing is as follows:

[0252] Lithocholic acid is mixed with 3 to 7 times its mass of maltodextrin to obtain primary premix A; lithochlic acid is mixed with 4 to 6 times its mass of maltodextrin; lithochlic acid is mixed with 5 times its mass of maltodextrin; the premixing time in the primary premixing process is 5 min to 20 min; the premixing time in the primary premixing process is 10 min to 15 min.

[0253] Mix the primary premix A with half the mass of the remaining maltodextrin to obtain the secondary premix B; the premixing time in the secondary premixing process is 10 min to 30 min; the premixing time in the secondary premixing process is 15 min to 25 min; the premixing time in the secondary premixing process is 20 min.

[0254] The secondary premix B, the remaining maltodextrin, and microcrystalline cellulose are mixed together to obtain the basic premix C; the premixing time in the basic premixing process is 15 min to 35 min; the premixing time in the basic premixing process is 20 min to 30 min; the premixing time in the basic premixing process is 25 min.

[0255] During the three-stage premixing process, the machine must be stopped after each stage of mixing is completed. At least two points should be taken from the upper part (1 / 3 of the distance from the top), the middle part (1 / 2 of the distance from the top), and the lower part (1 / 3 of the distance from the bottom) of the mixing chamber, with a total of at least six points. The lithocholic acid content at each point should be detected by HPLC, and the relative standard deviation (RSD) should be calculated. Only when the RSD is ≤8% can the next step be carried out.

[0256] (S02) After adding chicken liver powder to the basic premix C in step (S01) and mixing, add tocopherol and mix, and then sprinkle in silica and mix to obtain lithocholic acid additive powder; the mixing time after adding chicken liver powder in step (S02) is 5 min to 10 min, the mixing time after adding tocopherol is 5 min to 10 min, and the mixing time after sprinkling in silica is 5 min to 15 min; the mixing time after adding chicken liver powder is 8 min, the mixing time after adding tocopherol is 8 min, and the mixing time after sprinkling in silica is 10 min; multiple samples are taken from the powder premix D in step (S02), and the lithocholic acid content is quickly detected by HPLC to ensure that the relative standard deviation of the content at each point is ≤8%, and the angle of repose of the powder premix D is measured to ensure that the angle of repose is ≤35°;

[0257] (S03) Dissolve fish hydrolysate and yeast extract in sunflower seed oil to obtain oil phase dissolved product;

[0258] (S04) The oil phase dissolved in step (S03) is added to the lithocholic acid additive powder in step (S02) by stirring and spraying to obtain lithocholic acid additive liquid.

[0259] The application of lithocholic acid additives for the management of feline obesity or metabolic syndrome involves mixing the lithocholic acid additives into the cat's daily diet according to the cat's weight. After continuous feeding for ≥4 weeks, the cat's weight, body condition score, body fat index, abdominal circumference, and serum metabolic indicators are measured. At the 4th week, 8th week, and every 8 weeks thereafter, the cat's weight, body condition score, body fat index, abdominal circumference, and serum metabolic indicators are measured. Serum metabolic indicators include fasting blood glucose, triglycerides, total cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol.

[0260] Before starting feeding, at 2 weeks after the start, at 4 weeks after the start, and every 4 weeks thereafter, measure the serum alanine aminotransferase and aspartate aminotransferase levels in cats. When either the serum alanine aminotransferase or aspartate aminotransferase continuously exceeds 1.5 times the upper limit of the detection reference range, or increases by more than 30% compared to the individual's baseline level, immediately halve the dose of the lithocholic acid additive or suspend feeding. After the liver function indicators return to normal, feeding can be restarted under monitoring at a dosing rate of 15 - 20 mg / (kg body weight·day);

[0261] Continuous feeding should result in at least one of the following: a decrease of ≥5% in the cat's body weight, BCS score, or abdominal circumference compared to the baseline value, and / or a decrease of ≥10% in the triglyceride or total cholesterol level compared to the baseline value;

[0262] During feeding, measure the serum alanine aminotransferase and aspartate aminotransferase in cats every 2 - 4 weeks. When either indicator increases by more than 10% compared to the baseline, reduce the dose or suspend feeding;

[0263] Monitor the relative abundance changes of lithocholic acid derivatives in cat feces; lithocholic acid derivatives include isolithocholic acid, 3 - oxolithocholic acid, 7 - ketolithocholic acid, 12 - ketolithocholic acid, isolithocholenic acid, taurocholate - conjugated lithocholic acid, or glycine - conjugated lithocholic acid;

[0264] The dosing rate of the lithocholic acid additive is 15 - 45 mg / (kg body weight·day); The dosing rate of the lithocholic acid additive is 20 - 40 mg / (kg body weight·day); The dosing rate of the lithocholic acid additive is 25 - 35 mg / (kg body weight·day); The dosing rate of the lithocholic acid additive is 30 mg / (kg body weight·day);

[0265] Use the HPLC method to detect the content of lithocholic acid in the lithocholic acid additive powder. It is qualified within the range of 95% - 105% of the labeled amount of 100 mg / g;

[0266] The RSD of the within - batch uniformity of the lithocholic acid additive powder is ≤10% to be qualified;

[0267] The moisture content in the lithocholic acid additive powder is ≤5.0% to be qualified;

[0268] The angle of repose of the lithocholic acid additive powder is ≤35° to be qualified;

[0269] Take three batches of lithocholic acid additive powder samples and place them under accelerated test conditions for 3 months. Sampling and testing are carried out at the end of the 0th, 1st, 2nd, and 3rd months. The decline rate of the lithocholic acid content ≤5.0% is qualified; The accelerated test conditions are a temperature of 38℃ - 42℃ and a relative humidity of 70% - 80%; The accelerated test conditions are a temperature of 40℃ and a relative humidity of 75%;

[0270] If the lithocholic acid additive powder passes the test, it will be repackaged. During the repackaging process, each package contains a single daily dose or an integer multiple thereof suitable for a 5kg cat. Each package contains 0.25g to 0.35g. Each package contains 0.3g. Repackaging is carried out in a clean room with an ambient humidity of ≤45% using an automatic powder packaging machine and heat-sealed. The packaging bags used for repackaging are made of aluminum-plastic composite film material. The repackaged products are placed in cartons and stored in an environment below 25℃, dry and protected from light, with a shelf life of 12 to 18 months. Samples of each batch of products are retained for no less than three full inspections, and the storage conditions are 25℃, dry, protected from light and sealed.

[0271] Example 1: A lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following components by weight: 5 parts lithocholic acid, 70 parts maltodextrin, 5 parts microcrystalline cellulose, 1 part chicken liver powder, 0.2 parts silicon dioxide, and 0.05 parts tocopherol.

[0272] Example 2: A lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following components by weight: 12 parts lithocholic acid, 85 parts maltodextrin, 18 parts microcrystalline cellulose, 4 parts fish hydrolysate, 0.8 parts silicon dioxide, and 0.18 parts tocopherol.

[0273] Example 3: A lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following components by weight: 8 parts lithocholic acid, 75 parts maltodextrin, 8 parts microcrystalline cellulose, 2 parts yeast extract, 0.4 parts silica, and 0.08 parts tocopherol.

[0274] Example 4: A lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following components by weight: 12 parts lithocholic acid, 85 parts maltodextrin, 18 parts microcrystalline cellulose, 4 parts chicken liver powder, 0.8 parts silicon dioxide, and 0.18 parts tocopherol.

[0275] Example 5: A lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following components by weight: 9 parts lithocholic acid, 78 parts maltodextrin, 10 parts microcrystalline cellulose, 2 parts fish hydrolysate, 0.5 parts silica, and 0.1 parts tocopherol.

[0276] Example 6: A lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following components by weight: 11 parts lithocholic acid, 82 parts maltodextrin, 15 parts microcrystalline cellulose, 4 parts yeast extract, 0.7 parts silica, and 0.15 parts tocopherol.

[0277] Example 7: A lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following components by weight: 10 parts lithocholic acid, 80 parts maltodextrin, 12 parts microcrystalline cellulose, 3 parts chicken liver powder, 0.6 parts silicon dioxide, and 0.12 parts tocopherol.

[0278] Example 8: A lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following components by weight: 10 parts lithocholic acid, 77 parts maltodextrin, 10 parts microcrystalline cellulose, 2 parts chicken liver powder, 0.8 parts silicon dioxide, and 0.2 parts tocopherol.

[0279] Example 9: A lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following components by weight: 5 parts lithocholic acid, 70 parts maltodextrin, 5 parts microcrystalline cellulose, 1 part chicken liver powder, 0.2 parts silicon dioxide, 0.05 parts tocopherol, 90 parts sunflower seed oil, 1 part fish hydrolysate, and 1 part yeast extract.

[0280] Example 10: A lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following components by weight: 12 parts lithocholic acid, 85 parts maltodextrin, 18 parts microcrystalline cellulose, 1 part chicken liver powder, 0.8 parts silicon dioxide, 0.18 parts tocopherol, 100 parts sunflower seed oil, 2 parts fish hydrolysate, and 2 parts yeast extract.

[0281] Example 11: A lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following components by weight: 8 parts lithocholic acid, 75 parts maltodextrin, 8 parts microcrystalline cellulose, 2 parts chicken liver powder, 0.4 parts silicon dioxide, 0.08 parts tocopherol, 92 parts sunflower seed oil, 1.2 parts fish hydrolysate, and 1.2 parts yeast extract.

[0282] Example 12: A lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following components by weight: 12 parts lithocholic acid, 85 parts maltodextrin, 18 parts microcrystalline cellulose, 1.4 parts chicken liver powder, 0.8 parts silicon dioxide, 0.18 parts tocopherol, 98 parts sunflower seed oil, 1.8 parts fish hydrolysate, and 1.8 parts yeast extract.

[0283] Example 13: A lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following components by weight: 9 parts lithocholic acid, 78 parts maltodextrin, 10 parts microcrystalline cellulose, 2 parts chicken liver powder, 0.5 parts silicon dioxide, 0.1 parts tocopherol, 90 parts sunflower seed oil, 1 part fish hydrolysate, and 1 part yeast extract.

[0284] Example 14: A lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following components by weight: 11 parts lithocholic acid, 82 parts maltodextrin, 15 parts microcrystalline cellulose, 1 part chicken liver powder, 0.7 parts silicon dioxide, 0.15 parts tocopherol, 100 parts sunflower seed oil, 2 parts fish hydrolysate, and 2 parts yeast extract.

[0285] Example 15: A lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following components by weight: 10 parts lithocholic acid, 80 parts maltodextrin, 12 parts microcrystalline cellulose, 2 parts chicken liver powder, 0.6 parts silicon dioxide, 0.12 parts tocopherol, 95 parts sunflower seed oil, 1.5 parts fish hydrolysate, and 1.5 parts yeast extract.

[0286] Example 16: A lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following components by weight: 10 parts lithocholic acid, 77 parts maltodextrin, 10 parts microcrystalline cellulose, 2 parts chicken liver powder, 0.8 parts silicon dioxide, 0.2 parts tocopherol, 97.8 parts sunflower seed oil, 1 part fish hydrolysate, and 1 part yeast extract.

[0287] Example 17: Lithocholic acid was obtained by enzymatically directed dehydroxylation of ursodeoxycholic acid as a starting substrate. The enzymatically directed dehydroxylation reaction of lithochlic acid includes the following steps:

[0288] (a01) Enzyme system preparation

[0289] (a01-1) Inoculate Clostridium hiranonis strain into enhanced Clostridium medium and culture anaerobically; Clostridium hiranonis strain preservation number is JCM16549; the culture temperature in step (a01-1) is 35℃ and the culture time is 60h.

[0290] (a01-2) Collect the bacterial cells by centrifugation of the culture medium from step (a01-1), then sonicate and centrifuge to obtain the supernatant, and obtain crude enzyme solution;

[0291] (a01-3) Add ammonium sulfate to the crude enzyme solution in step (a01-2) for fractional precipitation, and collect the precipitate with a saturation of 30% to 60%;

[0292] (a01-4) Dissolve the precipitate in step (a01-3) with sterile water, then purify it by DEAE-Sepharose FastFlow ion exchange chromatography, collect the fraction with 12α-hydroxyl removal activity, freeze dry it to obtain enzyme powder; in step (a01-4), the specific activity of the enzyme powder is not less than 15 U / mg protein.

[0293] (a02) Enzymatic reaction

[0294] (a02-1) Dissolve ursodeoxycholic acid (UDCA) with a purity ≥98% in a mixed solution of phosphate buffer and methanol to obtain a substrate solution; the concentration of the substrate solution in step (a02-1) is 50 mg / mL; the pH of the mixed solution in step (a02-1) is 6.6, and the volume ratio of phosphate buffer to methanol in the mixed solution is 3.5:1;

[0295] (a02-2) Add enzyme powder to the substrate solution in step (a02-1) and stir to react; the mass of enzyme powder added in step (a02-2) is 6% of the mass of the substrate solution; the reaction temperature in step (a02-2) is 36℃, the reaction time is 30h, and the pH of the reaction system is 6.7; the reaction in step (a02-2) is carried out in a constant temperature oscillating reactor.

[0296] (a03) Product purification

[0297] (a03-1) After the reaction in step (a02-2), an equal volume of ethyl acetate was added to the mixture for multiple extractions. The organic phases were combined and evaporated to dryness. The extractions in step (a03-1) were performed twice, and the evaporation was carried out by rotary evaporation.

[0298] (a03-2) Dissolve the evaporated product from step (a03-1) in hot ethanol and perform hot filtration, then allow the filtrate to stand and age; the volume of hot ethanol in step (a03-2) is 9 times the volume of the evaporated product; the aging temperature in step (a03-2) is 3℃, and the aging time is 18h; the hot ethanol in step (a03-2) is 95% hot ethanol;

[0299] (a03-3) The white crystals aged in step (a03-2) are filtered and washed, and then vacuum dried to obtain lithocholic acid LCA; the vacuum drying temperature in step (a03-3) is 35℃, the vacuum drying pressure is -0.1MPa, and the vacuum drying time is 9h; the purity of lithocholic acid LCA in step (a03-3) is ≥95%.

[0300] Example 18: This example is basically the same as Example 17, except that the culture temperature in step (a01-1) is 38℃ and the culture time is 36h; the pH of the mixed solution in step (a02-1) is 6.9, and the volume ratio of phosphate buffer to methanol in the mixed solution is 4.5:1; the enzyme powder added in step (a02-2) is 10% of the substrate solution mass; the reaction temperature in step (a02-2) is 37℃, the reaction time is 28h, and the pH of the reaction system is 6.9; the number of extractions in step (a03-1) is 5; the volume of hot ethanol in step (a03-2) is 11 times the volume of the evaporated product; the static aging temperature in step (a03-2) is 6℃ and the static aging time is 6h; the vacuum drying temperature in step (a03-3) is 45℃, the vacuum drying pressure is -0.09MPa, and the vacuum drying time is 3h.

[0301] Example 19: This example is basically the same as Example 17, except that the culture temperature in step (a01-1) is 37℃ and the culture time is 48h; the pH of the mixed solution in step (a02-1) is 6.7 and the volume ratio of phosphate buffer to methanol in the mixed solution is 4.2:1; the reaction temperature in step (a02-2) is 36.5℃ and the reaction time is 29h, and the pH of the reaction system is 6.8; the number of extractions in step (a03-1) is 3; the volume of hot ethanol in step (a03-2) is 10 times the volume of the evaporated product; the static aging temperature in step (a03-2) is 4.5℃ and the static aging time is 12h; and the vacuum drying temperature in step (a03-3) is 40℃, the vacuum drying pressure is -0.095MPa, and the vacuum drying time is 6h.

[0302] Example 20: The preparation of microcrystalline cellulose (MCC) includes the following steps:

[0303] (b01) Take bleached wood pulp with α-cellulose content ≥90%, dry it, and then pulverize it; in step (b01), the pulp is pulverized to a fiber length of 1mm to 2mm;

[0304] (b02) After adding hydrochloric acid solution to the pulverized material in step (b01) and stirring to react, it is quickly cooled to room temperature with ice water; the solid-liquid ratio of the pulverized material to the hydrochloric acid solution in step (b02) is 1:10; the concentration of the hydrochloric acid solution in step (b02) is 2N; the reaction temperature in step (b02) is 80℃ and the reaction time is 120min.

[0305] (b03) After filtering the cooling mixture from step (b02), take the filtrate and wash it with deionized water until the pH of the filtrate is 6.5;

[0306] (b04) Dry the filtrate after washing in step (b03) until the moisture content is ≤5%; the drying temperature in step (b04) is 55°C; the drying in step (b04) is carried out in an oven;

[0307] (b05) The dried material from step (b04) is pulverized by an air jet mill and then sieved to obtain microcrystalline cellulose (MCC) powder; in step (b05), it is sieved through an 180-mesh sieve; the d of the microcrystalline cellulose (MCC) powder in step (b05) 50 50μm~60μm, d 90 ≤120μm, bulk density is 0.28g / cm³ 3 ~0.32g / cm 3 The angle of repose is ≤33°.

[0308] Example 21: This example is basically the same as Example 20, except that the solid-liquid ratio of the pulverized material to the hydrochloric acid solution in step (b02) is 1:20; the reaction temperature in step (b02) is 85°C and the reaction time is 80 min; (b03) the cooling mixture from step (b02) is filtered and the filtrate is taken, and then washed with deionized water until the pH of the filtrate is 7; the drying temperature in step (b04) is 60°C; and in step (b05) it is passed through a 200-mesh sieve.

[0309] Example 22: This example is basically the same as Example 20, except that the solid-liquid ratio of the pulverized material to the hydrochloric acid solution in step (b02) is 1:15; the reaction temperature in step (b02) is 82°C and the reaction time is 105 min; in step (b03), deionized water is used to wash until the pH of the filtrate is 6.8; the drying temperature in step (b04) is 57°C; and in step (b05), the filtrate is passed through a 190-mesh sieve.

[0310] Example 23: The preparation of maltodextrin includes the following steps:

[0311] (c01) Prepare corn starch slurry using food-grade corn starch, and adjust the pH of the corn starch slurry to 6.1 using NaOH solution; the dry matter content of the corn starch slurry in step (c01) is 25%; the concentration of the NaOH solution in step (c01) is 1.0M;

[0312] (c02) Add α-amylase to the corn starch slurry from step (c01), stir and liquefy, then cool; the α-amylase in step (c02) is a thermoresistant α-amylase; the enzyme activity of the α-amylase in step (c02) is ≥20000 U / g, and the amount of α-amylase added is 0.4‰ based on the dry weight of the corn starch; the liquefaction temperature in step (c02) is 90℃, and the liquefaction time is 60 min; the liquefaction endpoint in step (c02) is controlled by the degree of dextrinization (DE) value being between 10 and 15; the temperature in step (c02) is cooled to 55℃;

[0313] (c03) After adjusting the pH of the cooling solution in step (c02) to 4.5 using HCl solution, glucoamylase is added for saccharification; the concentration of the HCl solution in step (c03) is 1.0M; the enzyme activity of glucoamylase in step (c03) is ≥100000U / g, and the amount of glucoamylase added is 0.7‰ based on the dry weight of corn starch; the saccharification time in step (c03) is 2h.

[0314] (c04) After removing impurities by pressure filtration of the saccharified solution in step (c03), the filtrate is concentrated to a solid content of 35%–40%, and then spray-dried to obtain a white or light yellow powder of maltodextrin; the pressure filtration in step (c04) is completed using a plate and frame filter press; the spray drying in step (c04) is completed in a spray drying tower, and the atomizer frequency in the spray drying tower is 45Hz; the inlet temperature of the spray drying in step (c04) is 180℃, and the outlet temperature is 80℃; the moisture content of the maltodextrin in step (c04) is ≤5.0%, the degree of dextrinization (DE) is 12–14, and the average particle size is d 50 It is 50μm to 60μm.

[0315] Example 24: This example is basically the same as Example 23, except that (c01) food-grade corn starch is used to prepare corn starch slurry, and the pH of the corn starch slurry is adjusted to 6.3 using NaOH solution; the dry matter content of the corn starch slurry in step (c01) is 35%; the amount of α-amylase added is 0.6‰ based on the dry weight of the corn starch; the liquefaction temperature in step (c02) is 95℃, and the liquefaction time is 30 min; the liquefaction endpoint is controlled by the degree of dextrinization (DE) value being between 11 and 14; step (c02) (c02) Cool to 60℃; (c03) Adjust the pH of the cooling solution in step (c02) to 5 using HCl solution, and then add glucoamylase for saccharification; The amount of glucoamylase added is 0.9‰ based on the dry weight of corn starch; The saccharification time in step (c03) is 3h; In step (c04), concentrate the filtrate to a solid content of 36% to 39%; The atomizer frequency in the spray drying tower is 55Hz; The inlet temperature of the spray drying in step (c04) is 190℃ and the outlet temperature is 85℃.

[0316] Example 25: This example is basically the same as Example 23, except that the dry matter content of the corn starch slurry in step (c01) is 30%; the pH of the corn starch slurry is adjusted to 6.2; the amount of α-amylase added is 0.5‰ based on the dry weight of the corn starch; the liquefaction temperature in step (c02) is 92℃, and the liquefaction time is 40 min; the liquefaction endpoint in step (c02) is controlled by the degree of dextrinization (DE) value being 12-13; the temperature is cooled to 57℃ in step (c02); and HCl is used in step (c03). In solution adjustment step (c02), after cooling the solution to pH 4.8, glucoamylase is added for saccharification; the amount of glucoamylase added is 0.8‰ based on the dry weight of corn starch; the saccharification time in step (c03) is 2.5 h; in step (c04), the filtrate is concentrated to a solid content of 37%–38%; the atomizer frequency in the spray drying tower is 50 Hz; the inlet temperature of the spray dryer in step (c04) is 184℃, and the outlet temperature is 82℃; the degree of dextrinization (DE) is 13, and the average particle size is d. 50 It is 55μm.

[0317] Example 26: The preparation of chicken liver powder includes the following steps:

[0318] (d01) Cut chicken liver into pieces and pre-cook. Then add an equal amount of ice water and homogenize to obtain a homogenized liquid. The chicken liver used in step (d01) should be fresh chicken liver selected within 1-2 hours after slaughter. The chicken liver used in step (d01) should be chicken liver that has passed inspection and has had its fat tissue, blood vessels, and gallbladder removed. The chicken liver pieces in step (d01) should be 1.5cm in size. 3 ~2.5cm 3 The pre-cooking temperature in step (d01) is 80℃ and the pre-cooking time is 10 min; the crushing and homogenization in step (d01) is completed in a high-speed shear crusher; the homogenization speed in step (d01) is 4500 rpm and the homogenization time is 2 min.

[0319] (d02) After filtering the homogenized liquid from step (d01), the filtrate is spray-dried to obtain brown chicken liver powder; the filtration in step (d02) is completed through a 180-mesh stainless steel screen; the spray drying in step (d02) is completed in a spray drying tower, the atomizer frequency in the spray drying tower is 45Hz, and the atomization pressure is 0.4MPa; the inlet temperature of the spray drying in step (d02) is 160℃, and the outlet temperature is 80℃; the moisture content of the brown chicken liver powder in step (d02) is ≤6.0%, the crude protein content is ≥68%, and the crude fat content is ≤10%.

[0320] Example 27: This example is basically the same as Example 26, except that the pre-cooking temperature in step (d01) is 85°C and the pre-cooking time is 8 min; the homogenization speed in step (d01) is 5000 rpm and the homogenization time is 1 min; the filtration in step (d02) is completed through a 200-mesh stainless steel screen; the atomizer frequency in the spray drying tower is 55 Hz and the atomization pressure is 0.6 MPa; the inlet temperature of the spray drying in step (d02) is 170°C and the outlet temperature is 85°C.

[0321] Example 28: This example is basically the same as Example 26, except that the pre-cooking temperature in step (d01) is 82℃ and the pre-cooking time is 9min; the homogenization speed in step (d01) is 4800rpm and the homogenization time is 1.5min; the filtration in step (d02) is completed through a 190-mesh stainless steel screen; the atomizer frequency in the spray drying tower is 50Hz and the atomization pressure is 0.5MPa; the inlet temperature of the spray drying in step (d02) is 164℃ and the outlet temperature is 82℃.

[0322] Example 29: The preparation of silica includes the following steps,

[0323] (e01) Dilute industrial water glass with water; the modulus of the industrial water glass in step (e01) is 3.1; the SiO2 content in step (e01) is diluted to 8%;

[0324] (e02) Add sulfuric acid solution dropwise to the diluted solution in step (e01) under stirring until precipitation is complete; the stirring temperature in step (e02) is 25°C and the stirring speed is 250 rpm; the dropwise addition in step (e02) is uniform, and the mass concentration of the sulfuric acid solution is 15%; the pH of the system is maintained at 6 during the dropwise addition in step (e02).

[0325] (e03) Continue stirring and aging the solution after complete precipitation in step (e02); the aging time in step (e03) is 50 min;

[0326] (e04) Filter the aged solution from step (e03) by pressure filtration, and wash the resulting filter cake with deionized water until the Na+ in the wash solution is removed. + Concentration ≤100ppm; the filtration in step (e04) is plate and frame filtration, and the temperature of the deionized water is 55℃;

[0327] (e05) After drying the filter cake from step (e04), it is pulverized using an air jet mill and then sieved to obtain basic silica powder; the drying temperature in step (e05) is 100℃ and the drying time is 12h; the filter cake in step (e05) is sieved through a 190-mesh sieve.

[0328] (e06) The basic silica powder from step (e05) is placed in a fluidized bed and preheated. Then, using nitrogen as a carrier, the silane coupling agent is sprayed onto the surface of the fluidized powder. The preheating temperature in step (e06) is 140°C. The mass of the silane coupling agent in step (e06) is 0.5% of the mass of the basic silica powder. The silane coupling agent in step (e06) is a γ-aminopropyltriethoxysilane ethanol solution with a mass concentration of 8%. The spraying rate in step (e06) is 4 mL / min.

[0329] (e07) The material sprayed in step (e06) is subjected to heat treatment to obtain modified silica powder; the heat treatment temperature in step (e07) is 140℃ and the heat treatment time is 40min; the loose density of the modified silica powder in step (e07) is 0.12g / cm³, the angle of repose is ≤32°, and the specific surface area is 180m² / g.

[0330] Example 30: This example is basically the same as Example 29, except that the modulus of the industrial water glass in step (e01) is 3.5; the SiO2 content is diluted to 12% in step (e01); the stirring temperature in step (e02) is 30℃ and the stirring speed is 300 rpm; the pH of the system is maintained at 6.5 during the dropwise addition in step (e02); the aging time in step (e03) is 60 min; the temperature of the deionized water is 65℃; and the drying temperature in step (e05) is 105℃ and the drying time is 10 h. In step (e05), the silica powder is passed through a 210-mesh sieve; in step (e06), it is preheated to 150°C; in step (e06), the mass of the silane coupling agent is 1.5% of the mass of the base silica powder; the mass concentration of the γ-aminopropyltriethoxysilane ethanol solution is 12%; the spray rate in step (e06) is 6 mL / min; the heat treatment temperature in step (e07) is 150°C and the heat treatment time is 20 min; the loose density of the modified silica powder in step (e07) is 0.18 g / cm³ and the specific surface area is 240 m² / g.

[0331] Example 31: This example is basically the same as Example 29, except that the modulus of the industrial water glass in step (e01) is 3.3; the SiO2 content is diluted to 10% in step (e01); the stirring temperature in step (e02) is 28℃ and the stirring speed is 280 rpm; the pH of the system is maintained at 6.1 during the dropwise addition in step (e02); the aging time in step (e03) is 55 min; the temperature of the deionized water is 60℃; and the drying temperature in step (e05) is 102℃ and the drying time is 11 h. In step (e05), the silica powder is passed through a 200-mesh sieve; in step (e06), it is preheated to 145°C; in step (e06), the mass of the silane coupling agent is 1% of the mass of the base silica powder; the mass concentration of the γ-aminopropyltriethoxysilane ethanol solution is 10%; the spray rate in step (e06) is 5 mL / min; in step (e07), the heat treatment temperature is 145°C and the heat treatment time is 30 min; in step (e07), the loose density of the modified silica powder is 0.15 g / cm³ and the specific surface area is 210 m² / g.

[0332] Example 32: The preparation of tocopherol includes the following steps,

[0333] (f01) After melting the deodorized soybean oil distillate in a water bath, add ethanol and stir to dissolve; the water bath temperature in step (f01) is 50℃; the volume of ethanol added in step (f01) is 3 times the mass of the deodorized soybean oil distillate; the ethanol in step (f01) is 95% ethanol;

[0334] (f02) The solution in step (f01) is filtered, and the filtrate is subjected to two-stage molecular distillation to collect the fraction; the filtration in step (f02) is through a 0.4 μm filter membrane; the temperature of the first stage of molecular distillation in step (f02) is 100℃ and the vacuum degree of the first stage of molecular distillation is 8 Pa; the temperature of the second stage of molecular distillation in the two-stage molecular distillation is 140℃ and the vacuum degree of the second stage of molecular distillation is 6 Pa.

[0335] (f03) After concentrating the fraction from step (f02) under reduced pressure, add an equal volume of n-hexane for low-temperature crystallization; in step (f03), concentrate under reduced pressure to 1 / 3 of the original volume; the crystallization temperature in step (f03) is -20℃, and the crystallization time is 10h;

[0336] (f04) Filter the substance that has been crystallized in step (f03) to obtain crude tocopherol;

[0337] (f05) The crude tocopherol from step (f04) was subjected to silica gel column chromatography. The target component was collected according to the main peak and then rotary evaporated to obtain a light yellow oily tocopherol concentrate. The silica gel in step (f05) had a particle size of 200 mesh and a column diameter-to-height ratio of 1:13. The mobile phase in step (f05) was a mixture of petroleum ether and ethyl acetate with a volume ratio of 90:10. The total tocopherol content in the tocopherol concentrate of step (f05) was ≥75%, of which δ-tocopherol was ≥45%.

[0338] Example 33: This example is basically the same as Example 32, except that the water bath temperature in step (f01) is 60℃; the volume of ethanol added in step (f01) is 5 times the mass of soybean oil deodorized distillate; the filtration in step (f02) is through a 0.5μm filter membrane; the temperature of the first stage of molecular distillation in step (f02) is 120℃, and the vacuum degree of the first stage of molecular distillation is 12Pa; the temperature of the second stage of molecular distillation in step (f02) is 160℃, and the vacuum degree of the second stage of molecular distillation is 10Pa; the crystallization temperature in step (f03) is -18℃, and the crystallization time is 12h; the silica gel particle size in the silica gel column in step (f05) is 300 mesh, and the column diameter-to-height ratio of the silica gel column is 1:17; the volume ratio of petroleum ether to ethyl acetate is 95:5.

[0339] Example 34: This example is basically the same as Example 32, except that the water bath temperature in step (f01) is 55℃; the volume of ethanol added in step (f01) is 4 times the mass of soybean oil deodorized distillate; the filtration in step (f02) is through a 0.45μm filter membrane; the temperature of the first stage of molecular distillation in step (f02) is 110℃, the vacuum degree of the first stage of molecular distillation is 10Pa, the temperature of the second stage of molecular distillation in the two-stage molecular distillation is 150℃, and the vacuum degree of the second stage of molecular distillation is 8Pa; the crystallization temperature in step (f03) is -19℃, and the crystallization time is 11h; the silica gel particle size in the silica gel column in step (f05) is 250 mesh, the column diameter to height ratio of the silica gel column is 1:15; and the volume ratio of petroleum ether to ethyl acetate is 92:8.

[0340] Example 35: A method for preparing a lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following steps:

[0341] (S01) Lithocholic acid, maltodextrin and microcrystalline cellulose are fractionally premixed to obtain basic premix C; the fractional premixing in step (S01) is completed in a three-dimensional motion mixer;

[0342] The staged premixing in step (S01) is a three-stage premixing, and the three-stage premixing is as follows:

[0343] Lithocarpic acid was mixed with 3 times its mass of maltodextrin to obtain primary premix A; the premixing time in the primary premixing process was 5 min.

[0344] Mix the primary premix A with half the mass of the remaining maltodextrin to obtain the secondary premix B; the premixing time in the secondary premixing process is 10 min.

[0345] The secondary premix B, the remaining maltodextrin, and microcrystalline cellulose were mixed together to obtain the basic premix C; the premixing time in the basic premixing process was 15 min.

[0346] During the three-stage premixing process, the machine must be stopped after each stage of mixing is completed. At least two points should be taken from the upper part (1 / 3 of the distance from the top), the middle part (1 / 2 of the distance from the top), and the lower part (1 / 3 of the distance from the bottom) of the mixing chamber, with a total of at least six points. The lithocholic acid content at each point should be detected by HPLC, and the relative standard deviation (RSD) should be calculated. Only when the RSD is ≤8% can the next step be carried out.

[0347] (S02) After adding the masking agent to the basic premix C in step (S01) and mixing, add tocopherol and mix, then sprinkle in silica and mix to obtain lithocholic acid additive powder; the mixing time after adding the masking agent in step (S02) is 5 min, the mixing time after adding tocopherol is 5 min, and the mixing time after sprinkling in silica is 5 min; multiple samples are taken from the powder premix D in step (S02), and the lithocholic acid content is quickly detected by HPLC to ensure that the relative standard deviation of the content at each point is ≤8%, and the angle of repose of the powder premix D is measured to ensure that the angle of repose is ≤35°.

[0348] Example 36: This example is basically the same as Example 35, except that lithocholic acid is mixed with 7 times the mass of maltodextrin to obtain primary premix A; the premixing time in the primary premixing process is 20 min; the premixing time in the secondary premixing process is 30 min; the premixing time in the basic premixing process is 35 min; the mixing time after adding the masking agent in step (S02) is 10 min, the mixing time after adding tocopherol is 10 min, and the mixing time after sprinkling in silica is 15 min.

[0349] Example 37: This example is basically the same as Example 35, except that lithocholic acid is mixed with 5 times the mass of maltodextrin to obtain primary premix A; the premixing time in the primary premixing process is 10 min; the premixing time in the secondary premixing process is 20 min; the premixing time in the basic premixing process is 25 min; the mixing time after adding the masking agent in step (S02) is 8 min, the mixing time after adding tocopherol is 8 min, and the mixing time after sprinkling silica is 10 min.

[0350] Example 38: A method for preparing a lithocholic acid additive for the management of obesity or metabolic syndrome in cats, comprising the following steps:

[0351] (S01) Lithocholic acid, maltodextrin and microcrystalline cellulose are fractionally premixed to obtain basic premix C; the fractional premixing in step (S01) is completed in a three-dimensional motion mixer;

[0352] The staged premixing in step (S01) is a three-stage premixing, and the three-stage premixing is as follows:

[0353] Lithocarpic acid was mixed with 3 times its mass of maltodextrin to obtain primary premix A; the premixing time in the primary premixing process was 5 min.

[0354] Mix the primary premix A with half the mass of the remaining maltodextrin to obtain the secondary premix B; the premixing time in the secondary premixing process is 10 min.

[0355] The secondary premix B, the remaining maltodextrin, and microcrystalline cellulose were mixed together to obtain the basic premix C; the premixing time in the basic premixing process was 15 min.

[0356] During the three-stage premixing process, the machine must be stopped after each stage of mixing is completed. At least two points should be taken from the upper part (1 / 3 of the distance from the top), the middle part (1 / 2 of the distance from the top), and the lower part (1 / 3 of the distance from the bottom) of the mixing chamber, with a total of at least six points. The lithocholic acid content at each point should be detected by HPLC, and the relative standard deviation (RSD) should be calculated. Only when the RSD is ≤8 can the next step be carried out.

[0357] (S02) After adding chicken liver powder to the basic premix C in step (S01) and mixing, add tocopherol and mix, and then sprinkle in silica and mix to obtain lithocholic acid additive powder; the mixing time after adding chicken liver powder in step (S02) is 5 min, the mixing time after adding tocopherol is 5 min, and the mixing time after sprinkling in silica is 5 min; multiple samples are taken from the powder premix D in step (S02), and the lithocholic acid content is quickly detected by HPLC to ensure that the relative standard deviation of the content at each point is ≤8%, and the angle of repose of the powder premix D is measured to ensure that the angle of repose is ≤35°;

[0358] (S03) Dissolve fish hydrolysate and yeast extract in sunflower seed oil to obtain oil phase dissolved product;

[0359] (S04) The oil phase dissolved in step (S03) is added to the lithocholic acid additive powder in step (S02) by stirring and spraying to obtain lithocholic acid additive liquid.

[0360] Example 39: This example is basically the same as Example 38, except that lithocholic acid is mixed with 7 times the mass of maltodextrin to obtain primary premix A; the premixing time in the primary premixing process is 20 min; the premixing time in the secondary premixing process is 30 min; the premixing time in the basic premixing process is 35 min; the mixing time after adding chicken liver powder in step (S02) is 10 min, the mixing time after adding tocopherol is 10 min, and the mixing time after sprinkling in silica is 15 min.

[0361] Example 40: This example is basically the same as Example 38, except that lithocholic acid is mixed with 5 times the mass of maltodextrin to obtain primary premix A; the premixing time in the primary premixing process is 10 min; the premixing time in the secondary premixing process is 20 min; the premixing time in the basic premixing process is 25 min; the mixing time after adding chicken liver powder in step (S02) is 8 min, the mixing time after adding tocopherol is 8 min, and the mixing time after sprinkling in silica is 10 min.

[0362] Example 41: Application of lithocholic acid additive for the management of feline obesity or metabolic syndrome. The lithocholic acid additive was mixed into the cat's daily diet according to the cat's weight. After continuous feeding for ≥4 weeks, the cat's weight, body condition score, body fat index, abdominal circumference and serum metabolic indicators were measured. The cat's weight, body condition score, body fat index, abdominal circumference and serum metabolic indicators were measured at the 4th week, 8th week and every 8 weeks thereafter. The serum metabolic indicators included fasting blood glucose, triglycerides, total cholesterol, high-density lipoprotein cholesterol and low-density lipoprotein cholesterol.

[0363] Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in cats should be measured before feeding begins, at week 2 after feeding begins, at week 4 after feeding begins, and every 4 weeks thereafter. If either ALT or AST level remains above 1.5 times the upper limit of the reference range or increases by more than 30% from the individual’s baseline level, the lithocholic acid additive dosage should be halved or feeding should be suspended immediately. Once liver function indicators return to normal, feeding can be restarted under monitoring at a dosage of 15 mg / (kg body weight·day).

[0364] Continuous feeding should result in a decrease of at least 5% in one of the following from baseline: body weight, BCS score, or abdominal circumference; and / or a decrease of at least 10% in triglyceride or total cholesterol levels from baseline.

[0365] During feeding, the cat's serum alanine aminotransferase and aspartate aminotransferase were tested every 2 weeks. If either indicator increased by more than 10% from the baseline, the dose was reduced or feeding was suspended.

[0366] Monitoring the relative abundance changes of lithocholic acid derivatives in cat feces; lithocholic acid derivatives include isolithocholic acid, 3-oxolithocholic acid, 7-ketolithocholic acid, 12-ketolithocholic acid, isolithocholenic acid, taurocholic acid conjugated lithocholic acid, or glycine conjugated lithocholic acid;

[0367] The added dose of the lithocholic acid additive is 15 mg / (kg body weight·day);

[0368] Using the HPLC method to detect the content of lithocholic acid in the lithocholic acid additive powder, it is qualified within the range of 95% - 105% of the labeled amount of 100 mg / g;

[0369] The within-batch uniformity RSD of the lithocholic acid additive powder ≤ 10% is qualified;

[0370] The moisture content in the lithocholic acid additive powder ≤ 5.0% is qualified;

[0371] The angle of repose of the lithocholic acid additive powder ≤ 35° is qualified;

[0372] Take three batches of lithocholic acid additive powder samples, place them under accelerated test conditions for 3 months, and take samples for testing at the end of the 0th, 1st, 2nd, and 3rd months. The decline rate of the lithocholic acid content ≤ 5.0% is qualified; the accelerated test conditions are a temperature of 38°C and a relative humidity of 70%;

[0373] If the lithocholic acid additive powder passes the inspection, it will be sub-packaged; during the sub-packaging process, each packaging bag contains the single-day dose suitable for a 5 kg body weight cat or an integer multiple of the dose; each packaging bag contains 0.25 g during the sub-packaging process; the sub-packaging is carried out in a clean room with an environmental humidity ≤ 45% using an automatic powder packaging machine and sealed with heat sealing; the packaging bags used for sub-packaging are made of aluminum-plastic composite film materials; the sub-packaged products are placed in cartons and stored under environmental conditions of below 25°C, dry, and light-proof, with a shelf life of 12 months; each batch of products is留样 not less than three full inspection amounts and stored under conditions of 25°C, dry, light-proof, and sealed.

[0374] Example 42: This example is basically the same as Example 41, except that after the liver function indicators return to normal, feeding can be restarted under monitoring at an added dose of 20 mg / (kg body weight·day); during the feeding period, the serum alanine aminotransferase and aspartate aminotransferase of the cat are detected every 4 weeks, and when any index increases by more than 10% compared to the baseline, the dose is reduced or feeding is suspended; the added dose of the lithocholic acid additive is 45 mg / (kg body weight·day); the accelerated test conditions are a temperature of 42°C and a relative humidity of 80%; each packaging bag contains 0.35 g during the sub-packaging process; the shelf life is 18 months.

[0375] Example 43: This example is basically the same as Example 41, except that after liver function indicators return to normal, feeding can be restarted at an added dose of 18 mg / (kg body weight·day) under monitoring; during feeding, the cat's serum alanine aminotransferase and aspartate aminotransferase are tested every 3 weeks, and the dose is reduced or feeding is suspended when either indicator increases by more than 10% from the baseline; the added dose of lithocholic acid additive is 30 mg / (kg body weight·day); the accelerated test conditions are 40°C and 75% relative humidity; each package contains 0.3 g during the packaging process; the shelf life is 15 months.

[0376] Example 44: Preparation and quality control of lithocholic acid (LCA) additives for the management of obesity or metabolic syndrome in cats.

[0377] Lithocholic acid raw powder LCA, HPLC purity ≥95%. Lithocholic acid LCA is a hydrophobic bile acid with extremely low solubility in water and a melting point of about 186℃. It is suitable for the process route of powder dispersion or dissolution in an organic phase before dispersion.

[0378] Carriers and additives: maltodextrin, microcrystalline cellulose (MCC), silica, chicken liver powder, fish hydrolysate, yeast extract; microcrystalline cellulose (MCC) is safe and suitable as a feed additive carrier, and silica is a commonly used anti-caking and flow aid.

[0379] Antioxidants: Mixed tocopherols at 0.05%–0.2% are used for lipid protection in the formulation;

[0380] The product comprises the following components by weight: 10 parts lithocholic acid, 77 parts maltodextrin, 10 parts microcrystalline cellulose, 2 parts chicken liver powder, 0.8 parts silicon dioxide, and 0.2 parts tocopherol.

[0381] Label: Each gram contains 100mg of LCA, for easy measurement by body weight, such as 30mg / kg / day;

[0382] Preparation process:

[0383] (1) Sieving: All powder raw materials are sieved through a 60-mesh sieve;

[0384] (2) Stepwise mixing: The mixture is scaled up stepwise from 1:4 to 1:10 to the whole batch, with each step lasting no less than 5 minutes. Chicken liver powder and silicon dioxide are added to the last step, and tocopherol is added as well.

[0385] (3) In-process control: ≥10 samples are taken within the batch, and the content is determined by HPLC. The batch RSD ≤10% is required before repackaging.

[0386] (4) Packaging and sealing: Pack into small bags according to the indicated amount, such as 5kg body weight / bag·day, heat seal, batch number and expiration date label;

[0387] (5) Storage: Store in a dry, dark and sealed place at 25°C;

[0388] Quality control and stability:

[0389] For content determination, HPLC or UPLC external standard method is used: the release standard is 95% to 110% of the labeled amount, and the intra-batch RSD is ≤10%;

[0390] Stability: Long-term conditions of 25℃ and 60%RH and accelerated conditions of 40℃ and 75%RH are set. 30℃ and 65%RH can be set as intermediate conditions. The LCA content decreases by ≤5% and the sensory, moisture, clumping and microbial properties all meet the standards. Stability is judged as follows:

[0391] HPLC chromatographic conditions for LCA content determination:

[0392] Chromatographic column: reversed-phase C18, 4.6×250mm, 5μm or equivalent; column temperature: 35℃; flow rate: 1.0mL / min; injection volume: 10μL; detection: DAD or UV 210nm;

[0393] The mobile phase used was a gradient method: Phase A = methanol:acetonitrile:water in a volume ratio of 1:1:3 + 1 mM ammonium acetate + 0.1% acetic acid; Phase B = methanol:acetonitrile in a volume ratio of 1:1 + 0.1% acetic acid; the linear gradient was from 0 to 30 min, with Phase B from 40% to 95%, used to separate LCA and related bile acids; the total flow rate of the mobile phase was kept constant at 1.0 mL / min; after the gradient was completed, the mobile phase was equilibrated with the initial mobile phase for 10 min before the next injection;

[0394] System applicability: RSD of peak area for 5 consecutive needles ≤ 2.0%, theoretical plate number ≥ 5000, tailing factor 1.0~1.5;

[0395] Sample pretreatment: The sample was dissolved in methanol, sonicated for 10 min, filtered through a 0.22 μm PTFE membrane, and then injected.

[0396] Linearity and system suitability: The linear range is 5–200 μg / mL, R² ≥ 0.999; with repeated injection of the test solution, n=5, peak area RSD ≤ 2.0%, theoretical plate number ≥ 5000, tailing factor ≤ 1.5, and recovery rate 95%–105%.

[0397] Variations: Liquid or paste additives, added dropwise, mixed in, or used as a base for sauce packets;

[0398] The product comprises the following components by weight: 10 parts lithocholic acid, 77 parts maltodextrin, 10 parts microcrystalline cellulose, 2 parts chicken liver powder, 0.8 parts silicon dioxide, 0.2 parts tocopherol, 97.8 parts sunflower seed oil, 1 part fish hydrolysate, and 1 part yeast extract.

[0399] Usage: Add by dose according to body weight, so that the daily intake of LCA is approximately 30 mg / kg; uniformity is judged by sampling at ≥10 points within the batch, with RSD ≤10%.

[0400] Sample retention or storage sample management: For each batch of products, sample retention and storage sample management shall be carried out in accordance with the company's SOP. The amount of finished product sample retained shall not be less than the amount required to complete two full inspections, which shall be used for accelerated and long-term sample retention respectively. The sample label shall indicate the product name, batch number, specifications, sampler, date and storage conditions. The sample shall be kept throughout the shelf life indicated on the finished product label, and it is recommended to continue to keep it for no less than 12 months after the shelf life ends. The storage conditions for the sample shall be the same as those for the finished product, or sealed at 25°C away from light.

[0401] Experimental Example: Nutritional Intervention Trial for Obese Cats

[0402] Feeding method: Mix the lithocholic acid additive into the daily food according to body weight. The target dose is 30 mg / kg / day, and it is permissible to make minor adjustments within the range of 15 to 45 mg / kg / day, with 25 to 35 mg / kg / day being preferred.

[0403] Product Form and Preparation: Lithocholic acid (LCA) raw material is preferably selected with an HPLC purity of ≥95%. Given its hydrophobic nature and near insolubility in water, but good solubility and high melting point in organic solvents such as ethanol, the formulation and process are mainly based on solid premixing and powder dispersion. Additives can be used as inert carriers such as maltodextrin, microcrystalline cellulose, rice husk powder, or silica for low-content dilution and rheological improvement, thereby achieving stable feeding by weight in factory or home mixed feeding scenarios. To improve palatability and stability, fish oil, vegetable oil, and tocopherol antioxidant strategies can be combined in the oil phase. Under the premise of adhering to carrier safety, process sequence, and labeling specifications, LCA can be stably, uniformly, and measurably embedded in daily diets, meeting the productization requirements of this invention of weight-based measurement, ease of implementation, and ease of industrialization.

[0404] Safety monitoring: Test ALT, AST and ALP every 2 to 4 weeks; reduce the dose or stop feeding if any of the indicators increases by more than 10% from the baseline.

[0405] Metabolic endpoints: Measure cat body weight, BCS, FBMI, chest and abdominal circumference, as well as GLU, TG, TC, LDL and TP;

[0406] Body fat index (FBMI) is calculated using the following formula: FBMI = Chest circumference (cm) / (Hip length (cm)) 2 Among them, chest circumference is the circumference of the fullest part behind the xiphoid process, and hind limb length is the distance from the greater trochanter of the femur to the calcaneal tuberosity;

[0407] Antioxidant and oxidative stress: Significantly increased GSH-Px content was observed, with T-AOC and SOD showing an increasing trend, and MDA showing a decreasing trend; these indicators are used to reflect the level of antioxidant defense and lipid peroxidation.

[0408] Immune function and inflammatory factors: Elevated serum IgA, IgG and IgM levels were observed; in terms of the inflammatory spectrum, IL-6, IL-1β and TNF-α showed a decreasing trend, while IL-10 showed an increasing trend.

[0409] Inclusion and grouping: Adult domestic cats that meet the criteria for obesity, with BCS ≥ 6 or FBMI exceeding the standard, are included in the obesity group OB, n=12; a healthy control group CON is included simultaneously as a reference, with only baseline comparison and no intervention; BCS is assessed according to the WSAVA nine-point scale, and FBMI is calculated based on chest circumference and hind limb length.

[0410] Diet and dosage: The OB group was given lithocholic acid additive at 30 mg / kg / d on the basis of the original diet. The specific formula in Example 8 was selected, and individual fine-tuning was allowed within the range of 15-45 mg / kg / d, with 25 mg / kg / d to 35 mg / kg / d being preferred.

[0411] Period and time points: 6 weeks of intervention; sampling time points were W0 baseline, W3, and W6;

[0412] Primary metabolic endpoints: body weight, BCS, FBMI, chest circumference, and abdominal circumference; metabolic biochemistry: GLU, TG, TC, LDL, and TP.

[0413] Antioxidant and oxidative stress endpoints: serum GSH-Px, SOD, and T-AOC were detected by enzymatic or colorimetric methods; MDA was determined by the TBARS method.

[0414] Immune or inflammatory endpoints: Serum IgA, IgG, IgM, IL-6, IL-1β, TNF-α, and IL-10 were quantified using a cat ELISA kit;

[0415] Safety monitoring: ALT, AST, ALP, TBIL, Alb, Glob, A and G should be retested every 2 to 4 weeks; when ALT or AST increases by more than 10% from baseline, the dose should be reduced to 25 mg / kg / day or the medication should be temporarily discontinued, and resumed after the retest shows that the target has been met;

[0416] Results Summary and Interpretation: Compared with baseline, the OB group achieved significant decreases in body weight, FBMI, BCS, and abdominal circumference at W6; GLU, TG, TC, and LDL decreased, and LDL improved; the antioxidant spectrum showed a significant increase in GSH-Px, an upward trend in T-AOC and SOD, and a decrease in MDA; the immune and inflammatory spectrum showed increases in IgA, IgG, and IgM, decreases in IL-6, IL-1β, and TNF-α, and an increase in IL-10; safety was manageable within the monitoring window.

[0417] Statistics: Paired t-test or repeated measures ANOVA; significance threshold p < 0.05; directional targets are determined based on differences from baseline or differences compared to controls;

[0418] Corresponding image: Figure 1 CONvsOB fecal bile acid profile; Figure 2 Graphs showing changes in body weight, food intake, and daily weight gain; Figure 3 A graph showing changes in body size and blood lipids / glucose levels; Figure 4 A graph showing changes in antioxidant and immune inflammation; Figure 5 This is a graph showing changes in liver function and protein metabolism.

[0419] It is evident that, compared to healthy cats, obese cats exhibit altered fecal bile acid profiles. Specifically, the profiles of LCA and its microbial derivatives, including intestinal microbial derivatives such as Lithocholic acid, 7-Ketolithocholic acid, 12-Ketolithocholic acid, Isolithocholic acid, Taurolithocholic acid, Dehydrolithocholic acid, Glycolithocholic acid, and Isoallolithocholic acid, have changed, suggesting a link between microbial metabolism and immune and metabolic mechanisms.

[0420] Supplementing with 30mg / kg / day of lithocholic acid can reduce weight, FBMI, BCS, and waist circumference, and improve blood sugar and blood lipids. Its metabolic rationale lies in the fact that LCA, as a strong TGR5 agonist, can promote GLP-1 secretion and improve energy and glucose and lipid metabolism.

[0421] No meal replacement or prescription diet replacement is required; it can be directly mixed into the feed, making it easy to implement in home settings. It significantly increases GSH-Px, with T-AOC and SOD showing an upward trend and MDA showing a downward trend, indicating an overall improvement in antioxidant capacity and a reduction in lipid peroxidation. IgA, IgG, and IgM levels show a recovery trend, indicating an enhanced humoral immune response. In terms of the inflammatory spectrum, IL-6, IL-1β, and TNF-α decrease while IL-10 increases, indicating an improvement in low-grade inflammation.

Claims

1. A lithocholic acid additive for the management of obesity or metabolic syndrome in cats, characterized by: It includes the following components in parts by weight. Lithocholic acid 5-15 parts, maltodextrin 70-88 parts, microcrystalline cellulose 5-20 parts, flavor masking agent 1-5 parts, silicon dioxide 0.2-1.0 parts, tocopherol 0.05-0.2 parts.

2. The lithocholic acid additive for the management of feline obesity or metabolic syndrome according to claim 1, characterized in that: The masking agent is selected from at least one of chicken liver powder, fish hydrolysate, or yeast extract.

3. The lithocholic acid additive for the management of feline obesity or metabolic syndrome according to claim 1, characterized in that: It also includes the following components by weight: 90-100 parts sunflower seed oil, 1-2 parts fish hydrolysate, and 1-2 parts yeast extract.

4. The lithocholic acid additive for the management of feline obesity or metabolic syndrome according to claim 1, characterized in that: The lithocholic acid was obtained by enzymatically dehydroxylating ursodeoxycholic acid as a starting substrate. The enzymatic directional dehydroxylation reaction of lithocholic acid Includes the following steps, (a01) Enzyme system preparation (a01-1) Take Clostridium hiranonis strain and inoculate it into enhanced Clostridium medium for anaerobic culture; (a01-2) Collect the bacterial cells by centrifugation of the culture medium from step (a01-1), then sonicate and centrifuge to obtain the supernatant, and obtain crude enzyme solution; (a01-3) Add ammonium sulfate to the crude enzyme solution in step (a01-2) for fractional precipitation, and collect the precipitate with a saturation of 30% to 60%; (a01-4) Dissolve the precipitate in step (a01-3) with sterile water, then purify it by DEAE-Sepharose Fast Flow ion exchange chromatography, collect the fraction with 12α-hydroxyl removal activity, freeze dry it to obtain enzyme powder; (a02) Enzymatic reaction (a02-1) Dissolve ursodeoxycholic acid (UDCA) with a purity ≥98% in a mixed solution of phosphate buffer and methanol to obtain a substrate solution; (a02-2) Add enzyme powder to the substrate solution in step (a02-1) and stir to react; (a03) Product purification (a03-1) After the reaction in step (a02-2), add an equal volume of ethyl acetate to the mixture and extract multiple times. Combine the organic phases and evaporate to dryness. (a03-2) Dissolve the evaporated product from step (a03-1) in hot ethanol and perform hot filtration, then allow the filtrate to stand and age. (a03-3) The white crystals aged in step (a03-2) are filtered and washed, and then dried under vacuum to obtain lithocholic acid LCA.

5. The lithocholic acid additive for the management of feline obesity or metabolic syndrome according to claim 4, characterized in that: The pH of the mixed solution in step (a02-1) is 6.6 to 6.9, and the volume ratio of phosphate buffer to methanol in the mixed solution is 3.5 to 4.5:

1. In step (a02-2), the enzyme powder is added at a mass of 6% to 10% of the substrate solution mass. In step (a03-2), the volume of hot ethanol is 9 to 11 times the volume of the evaporated product.

6. A method for preparing lithocholic acid additives for the management of obesity or metabolic syndrome in cats, characterized in that: Including the following step, (S01) Lithocholic acid, maltodextrin and microcrystalline cellulose are fractionated and premixed to obtain basic premix C; The staged premixing in step (S01) is a three-stage premixing, wherein the three-stage premixing is as follows: Lithocarpic acid is mixed with 3 to 7 times its mass of maltodextrin to obtain primary premix A; the premixing time in the primary premixing process is 5 min to 20 min. The primary premix A is mixed with half the mass of the remaining maltodextrin to obtain the secondary premix B; the premixing time in the secondary premixing process is 10 min to 30 min. The secondary premix B, the remaining maltodextrin, and microcrystalline cellulose were mixed together to obtain the basic premix C; The premixing time in the basic premixing process is 15 min to 35 min; During the three-stage premixing process, the machine must be stopped after each stage of mixing is completed. Multiple samples are taken from the mixing chamber, and the lithocholic acid content at each point is detected by HPLC. The relative standard deviation (RSD) is calculated, and the RSD must be ≤8% before proceeding to the next step. (S02) After adding the masking agent to the basic premix C in step (S01) and mixing, add tocopherol and mix, then sprinkle in silica and mix to obtain lithocholic acid additive powder.

7. A method for preparing lithocholic acid additives for the management of obesity or metabolic syndrome in cats, characterized in that: Including the following step, (S01) Lithocholic acid, maltodextrin and microcrystalline cellulose are fractionated and premixed to obtain basic premix C; The staged premixing in step (S01) is a three-stage premixing, wherein the three-stage premixing is as follows: Lithocarpic acid is mixed with 3 to 7 times its mass of maltodextrin to obtain primary premix A; the premixing time in the primary premixing process is 5 min to 20 min. The primary premix A is mixed with half the mass of the remaining maltodextrin to obtain the secondary premix B; the premixing time in the secondary premixing process is 10 min to 30 min. The secondary premix B, the remaining maltodextrin, and microcrystalline cellulose were mixed together to obtain the basic premix C; The premixing time in the basic premixing process is 15 min to 35 min; During the three-stage premixing process, the machine must be stopped after each stage of mixing is completed. Multiple samples are taken from the mixing chamber, and the lithocholic acid content at each point is detected by HPLC. The relative standard deviation (RSD) is calculated, and the RSD must be ≤8% before proceeding to the next step. (S02) After adding chicken liver powder to the basic premix C in step (S01) and mixing, add tocopherol and mix, then sprinkle in silicon dioxide and mix to obtain lithocholic acid additive powder. (S03) Dissolve fish hydrolysate and yeast extract in sunflower seed oil to obtain oil phase dissolved product; (S04) The oil phase dissolved in step (S03) is added to the lithocholic acid additive powder in step (S02) by stirring and spraying to obtain lithocholic acid additive liquid.

8. The application of lithocholic acid additives for the management of obesity or metabolic syndrome in cats, characterized by: Lithocholic acid additives were mixed into the cats' daily diet according to the cats' weight and fed continuously for ≥4 weeks. The cats' weight, body condition score, body fat index, abdominal circumference and serum metabolic indicators were then tested. Continuous feeding should result in a decrease of at least 5% in one of the following from baseline: body weight, BCS score, or abdominal circumference; and / or a decrease of at least 10% in triglyceride or total cholesterol levels from baseline. During feeding, serum alanine aminotransferase and aspartate aminotransferase in cats should be tested every 2 to 4 weeks. If either indicator increases by more than 10% from baseline, the dosage should be reduced or feeding should be suspended.

9. The application of the lithocholic acid additive for the management of feline obesity or metabolic syndrome according to claim 8, characterized in that: The dosage of the lithocholic acid additive is 15-45 mg / (kg body weight·day); the cat's weight, body condition score, body fat index, abdominal circumference and serum metabolic indicators are measured every 8 weeks after the 4th week, 8th week and thereafter. Before starting feeding, at 2 weeks after the start, at 4 weeks after the start, and every 4 weeks thereafter, the serum alanine aminotransferase and aspartate aminotransferase levels of the cats were measured. When any one of the serum alanine aminotransferase or aspartate aminotransferase indices continuously exceeded 1.5 times the upper limit of the detection reference range, or increased by more than 30% compared to the individual's baseline level, the dose of the lithocholic acid additive should be immediately halved or feeding should be suspended. After the liver function indices returned to normal, feeding could be restarted under monitoring at an addition dose of 15 - 20 mg / (kg body weight·day).

10. The application of the lithocholic acid additive for the management of feline obesity or metabolic syndrome according to claim 8, wherein: The content of lithocholic acid in the lithocholic acid additive powder is detected by HPLC method, and it is qualified within the range of 95% - 105% of the labeled amount of 100 mg / g; The RSD of the within - batch uniformity of the lithocholic acid additive powder ≤ 10% is qualified; The moisture content in the lithocholic acid additive powder ≤ 5.0% is qualified; The angle of repose of the lithocholic acid additive powder ≤ 35° is qualified; Three batches of lithocholic acid additive powder samples were taken and placed under accelerated test conditions for 3 months. Samples were taken and detected at the end of the 0th, 1st, 2nd, and 3rd months. The decline rate of the lithocholic acid content ≤ 5.0% is qualified; If the lithocholic acid additive powder passes the test, it is packaged; in the packaging process, each packaging bag contains a single - day dose for a cat with a body weight of 5 kg or an integer multiple of the dose; Each packaging bag contains 0.25 g - 0.35 g in the packaging process; The packaging is carried out in a clean room with an environmental humidity ≤ 45% using an automatic powder packaging machine and sealed by heat - sealing; the packaging bags used for packaging are made of aluminum - plastic composite film materials.

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