Chrysanthemum flower powder composition with fat-reducing effect and preparation method thereof

Abstract

This invention relates to a chrysanthemum powder composition with fat-reducing effects and its preparation method, belonging to the field of fermentation product preparation technology. The method includes the following steps: S1: Chrysanthemum powder, lotus leaf powder, and kudzu root powder are sieved separately and mixed to obtain a mixed powder; S2: The mixed powder is soaked in purified water, subjected to ultrasonic pretreatment, sterilized by enzyme inactivation, and then cooled to obtain a plant-based liquid; S3: Epigallocatechin gallate (EGCG) is added to the plant-based liquid based on the dry weight of the raw materials, followed by inoculation with fermentation bacteria, and the mixture is stirred and dispersed evenly to obtain a fermentation system; S4: The fermentation system is subjected to static micro-anaerobic fermentation, with stirring during the process, to obtain a fermentation broth; S5: The fermentation broth is refrigerated and allowed to stand, centrifuged and filtered, then concentrated at low temperature, freeze-dried, and sieved to finally obtain the chrysanthemum powder composition with fat-reducing effects. The chrysanthemum powder composition prepared by this invention has a high viable bacteria count, good stability, and a significant fat-reducing effect.

Description

Technical Field

[0001] This invention belongs to the field of fermentation product preparation technology, specifically, it relates to a chrysanthemum powder composition with fat-reducing effect and its preparation method. Background Technology

[0002] With high-fat, high-sugar diets and sedentary lifestyles becoming the norm, the incidence of obesity and related metabolic disorders continues to rise, becoming a significant threat to public health. Obesity not only affects physical appearance but also easily leads to abnormal glucose and lipid metabolism diseases such as hyperlipidemia and fatty liver. Therefore, safe and effective weight-loss functional foods have become a hot topic in research and development.

[0003] Chrysanthemum, lotus leaf, and kudzu root are traditional medicinal and edible ingredients. Chrysanthemum is rich in flavonoids and polyphenols, lotus leaf is rich in alkaloids and flavonoids, and kudzu root is rich in active ingredients such as puerarin. The combination of these three ingredients has potential lipid-lowering and fat-reducing effects. However, directly pulverizing them results in low dissolution rates and low bioavailability of the active ingredients. Meanwhile, the fat-reducing effect of a single plant ingredient is limited. Probiotics, on the other hand, can achieve fat reduction by regulating gut microbiota and improving lipid metabolism. Fermenting plant ingredients with probiotics can utilize microbial metabolism to enhance the bioavailability of active ingredients, achieving a synergistic fat-reducing effect between plant active ingredients and probiotics.

[0004] Existing weight-loss plant-based compound products are mostly prepared through simple mixing and decoction, which has problems such as insufficient dissolution of active ingredients, lack of targeting, and single weight-loss effect. Moreover, the combination of probiotics and plant raw materials is mostly directly added, which is easily affected by the components in the plant raw materials, resulting in low number of live bacteria, poor stability, and difficulty in exerting their synergistic effect.

[0005] Therefore, there is an urgent need to develop a chrysanthemum powder composition with fat-reducing effects and its preparation method. Summary of the Invention

[0006] The purpose of this invention is to provide a chrysanthemum powder composition with fat-reducing effect and its preparation method. By using low-temperature pulverization and ultrasonic-assisted solubilization pretreatment to improve the dissolution rate of plant active ingredients, and combining it with compound probiotics for micro-anaerobic fermentation, a synergistic fat-reducing effect of three plant active ingredients (chrysanthemum, lotus leaf, and kudzu root) and probiotics is achieved. The prepared chrysanthemum powder composition has a high number of live bacteria, good stability, and significant fat-reducing effect. It can also be prepared as a liquid beverage or compound powder to meet different consumption needs.

[0007] The objective of this invention can be achieved through the following technical solutions: A method for preparing a chrysanthemum powder composition with fat-reducing effects includes the following steps: S1: Take chrysanthemum powder, lotus leaf powder and kudzu root powder, sift them separately, and mix them to obtain a mixed powder; S2: Add the mixed powder to purified water and soak at 50-55℃, then perform ultrasonic solubilization pretreatment, and cool after enzyme inactivation sterilization to obtain plant-based liquid; S3: Based on the dry weight of the raw materials, add epigallocatechin gallate to the plant substrate liquid, then inoculate with fermentation strains. The total inoculation amount is 0.5-1.5‰. Stir and disperse evenly to obtain the fermentation system. S4: Allow the fermentation system to undergo static micro-anaerobic fermentation for 24-36 hours, stirring at low speed for 5-8 minutes every 1.5-2 hours, until the system pH reaches 3.6-4.0 and the viable cell count is ≥1×10⁻⁶. 9 CFU / g was used to obtain the fermentation broth; S5: After the fermentation liquid is refrigerated and allowed to stand for 1-2 hours, centrifuged and filtered, it is concentrated at low temperature to a solid content of 15%-20%, and then freeze-dried and sieved to obtain a chrysanthemum powder composition with fat-reducing effect.

[0008] Furthermore, a method for preparing a chrysanthemum powder composition with fat-reducing effects includes the following specific steps: S1: Take chrysanthemum powder, lotus leaf powder and kudzu root powder according to the weight parts, sift them separately, and mix them to obtain a mixed powder; S2: Add 8-10 times the amount of purified water to the mixed powder, soak at 50-55℃ for 30 minutes, then perform ultrasonic pretreatment, then heat at 90℃ for 10 minutes to inactivate enzymes and sterilize, and cool to 37±1℃ to obtain plant base liquid; S3: Based on the dry weight of the raw materials, add 1‰-3‰ epigallocatechin gallate (hereinafter referred to as EGCG) to the plant substrate liquid, then inoculate with fermentation strains. The total inoculation amount is 0.5-1.5‰. Stir and disperse evenly to obtain the fermentation system. S4: The fermentation system is statically micro-anaerobic at 37℃ for 24-36 hours, with low-speed stirring at 30 r / min for 5 minutes every 2 hours, until the pH of the system is 3.6-4.0 and the viable count is ≥1×10⁻⁶. 9 CFU / g was used to obtain the fermentation broth; S5: The fermentation broth is refrigerated at 0-4℃ and allowed to stand for 1-2 hours. After centrifugation and filtration, it is concentrated at low temperature to a solid content of 15%-20%. Then, it is freeze-dried and passed through an 80-mesh sieve to obtain a chrysanthemum powder composition with fat-reducing effect.

[0009] In the above technical solutions, the core active ingredients of chrysanthemum powder are total chrysanthemum flavonoids and chlorogenic acid. The principle is that flavonoids can inhibit pancreatic lipase activity, reduce the absorption of dietary fat in the intestines, and also have antioxidant and anti-inflammatory effects. It accounts for 70-90% of the main raw materials to ensure the basic content of flavonoids. The core active ingredients of lotus leaf powder are lotus leaf alkaloids and lotus leaf alkaloids. The principle is that lotus leaf alkaloids can inhibit the differentiation of adipocytes and lipid accumulation, while promoting intestinal peristalsis and reducing fat absorption. Its proportion is 10-20% to avoid the bitter taste caused by excessive lotus leaf alkaloid content, and to form a synergistic fat-reducing effect with chrysanthemum flavonoids. The core active ingredients of kudzu root powder are puerarin and starch. The principle is that puerarin can regulate glucose and lipid metabolism, lower blood sugar and reduce fat synthesis. Starch provides a fermentable carbon source for probiotics, promoting the growth and reproduction of probiotics. In addition, the colloidal properties of kudzu root powder can protect the survival of probiotics in the gastrointestinal tract. Its proportion is 1-10% to avoid excessive starch causing the fermentation system to become too viscous, affecting stirring and dissolved oxygen.

[0010] Further, in step S1, the mass ratio of chrysanthemum powder, lotus leaf powder, and kudzu root powder is 70-90:10-20:1-10.

[0011] Furthermore, in step S1, the temperature of the material during sieving is 0-5℃ to avoid degradation of plant active ingredients due to high temperature during the sieving process.

[0012] Furthermore, in step S1, the sieving process involves passing chrysanthemum powder and lotus leaf powder through a 40-mesh sieve, and kudzu root powder through a 60-mesh sieve.

[0013] In the above technical solution, the low temperature environment of 0-5℃ can inhibit the activity of polyphenol oxidase and lipase in chrysanthemum and lotus leaf, and prevent the active ingredients such as flavonoids and lotus leaf alkaloids from being enzymatically degraded or degraded in the subsequent process; chrysanthemum powder and lotus leaf powder are passed through a 40-mesh sieve, and kudzu root powder is passed through a 60-mesh sieve, which not only ensures the contact area between the raw material particles and water and improves the subsequent dissolution efficiency, but also avoids the subsequent filtration blockage caused by excessively fine particles.

[0014] Furthermore, in step S2, the ultrasonic-assisted dissolution pretreatment has an ultrasonic power of 200-300W, a temperature of 45-50℃, and a time of 20min.

[0015] In the above technical solution, a gentle temperature can enhance the permeability of plant cells, allowing purified water to penetrate into the intercellular spaces and initially dissolve water-soluble active ingredients such as puerarin and some flavonoids, without reaching the degradation threshold of the active ingredients. The cavitation effect of ultrasound generates microjets and shock waves, breaking down the cellulose and hemicellulose structure of plant cell walls, releasing lipid-soluble and bound active ingredients such as lotus leaf alkaloids and total chrysanthemum flavonoids into the solution, which can significantly improve the dissolution rate; a temperature of 45-50℃ helps reduce the viscosity of the cell sap and enhance the cavitation effect.

[0016] In the above technical solution, the treatment of heating at 90℃ for 10 minutes to inactivate enzymes and sterilize is a short-term high-temperature treatment that can completely inactivate endogenous enzymes in raw materials, avoiding enzymatic degradation of active ingredients in subsequent fermentation; at the same time, it kills molds, bacilli and other miscellaneous bacteria on the surface of raw materials, preventing miscellaneous bacteria from competing with probiotics for nutrients; the short-term high temperature is key, which can reduce the loss of heat-sensitive components such as flavonoids and alkaloids.

[0017] Furthermore, in step S3, the fermentation strain composition based on the dry weight of the raw materials is: Lactobacillus plantarum LP-Onlly 0.3-1.0‰ and Akkermansia AKK016 0.2-0.5‰.

[0018] In the above technical solution, EGCG is first added and dissolved to create an antioxidant microenvironment for probiotics, protecting the probiotic cell membranes from oxidative damage caused by polyphenols in the plant raw materials. The two strains are inoculated in a specific ratio to achieve symbiotic metabolism between probiotics. Lactobacillus plantarum LP-Onlly utilizes the starch in kudzu root powder and the polysaccharides in chrysanthemum powder, while Akkermansia muciniphila AKK016 utilizes the organic acids in lotus leaf powder and fermentation intermediates. This avoids nutrient competition and significantly increases the number of viable bacteria in the fermentation system. Stirring and dispersing ensure that the probiotics and EGCG are evenly distributed in the liquid, guaranteeing the uniformity of the fermentation system and avoiding incomplete fermentation caused by excessively high or low concentrations of local strains.

[0019] The selected dual-strain, *Lactobacillus plantarum* LP-Onlly, plays a key role in: regulating the intestinal flora structure, promoting the growth of beneficial bacteria, inhibiting harmful bacteria such as Firmicutes, and reducing intestinal fat absorption; simultaneously, it produces organic acids such as lactic acid and acetic acid, lowers intestinal pH, enhances the absorption of minerals such as calcium and magnesium, and promotes intestinal peristalsis; in addition, it can utilize the polysaccharides of chrysanthemum pollen and lotus leaf powder to produce short-chain fatty acids such as butyric acid and propionic acid, which can activate the AMPK pathway and promote fat breakdown; it accounts for a relatively high proportion of the main strain to ensure the basic viable count of the fermentation system.

[0020] Akkermansia mucinophilus AKK016 can degrade mucin in the intestinal mucus layer, providing nutrients for itself and Lactobacillus plantarum LP-Onlly, while enhancing intestinal barrier function and reducing fat accumulation in the intestine. It also significantly reduces body fat percentage and liver fat content in high-fat model mice. It forms a metabolic symbiosis with Lactobacillus plantarum LP-Onlly, further enhancing the fat reduction effect. In addition, this strain is strictly anaerobic and adapted to a micro-anaerobic fermentation environment, so the inoculum size is slightly lower than that of Lactobacillus plantarum LP-Onlly.

[0021] Furthermore, in step S4, the micro-anaerobic fermentation environment is an anaerobic culture environment with a CO2 volume fraction of 5%-8%.

[0022] In the above technical solution, the system is cultured in a micro-anaerobic fermentation environment at 37℃ and a CO2 volume fraction of 5%-8% for 24-36 hours. 37℃ is the optimal growth temperature for all compound probiotics, ensuring the metabolic activity of the strains. The micro-anaerobic environment with a CO2 volume fraction of 5%-8% can inhibit the residual growth of aerobic bacteria, while also meeting the metabolic needs of *Ackermania mutans* and *Lactobacillus plantarum*, promoting the production of fermentation products such as short-chain fatty acids and organic acids by the probiotics, and lowering the pH of the system to 3.6-4.0. The fermentation time of 24-36 hours ensures that the probiotics complete the logarithmic phase to the stationary phase of the growth curve, and the viable count reaches 1×10⁻⁶. 9 It contains CFU / g or higher and has fully converted plant active ingredients.

[0023] Stir at a low speed of 30 rpm for 5 minutes every 2 hours. The low speed stirring avoids disrupting the micro-anaerobic environment and solves the problem of nutrient stratification during static fermentation, allowing the nutrients in the liquid to come into full contact with the probiotics. Stirring promotes the release of a small amount of CO2 produced during fermentation, avoiding excessive local CO2 concentration that inhibits the metabolism of probiotics. The low speed of 30 rpm also prevents the probiotic cell membrane from being damaged by shear force.

[0024] Furthermore, in step S5, the freeze-drying process involves pre-freezing at -80°C for 14 hours, followed by freeze-drying at -45°C for 24 hours.

[0025] Furthermore, in step S5, the centrifugal filtration speed is 3000-4000 r / min, the time is 10-15 min, and the filtration uses a 200-mesh food-grade stainless steel filter screen.

[0026] In the above technical solution, the product is prepared by means of cold storage, centrifugal filtration, low temperature concentration, freeze drying and sieving. Low temperature reduces the metabolic activity of probiotics, puts the fermentation system in a dormant state, and avoids the loss of active ingredients due to excessive metabolism of probiotics in subsequent processing; at the same time, the static setting allows the insoluble residues in the liquid to settle naturally, improving the efficiency of subsequent filtration.

[0027] A chrysanthemum powder composition with fat-reducing effect prepared by the above preparation method.

[0028] The beneficial effects of this invention are: (1) In the technical solution of the present invention, the pretreatment process of low temperature sieving and ultrasonic solubilization effectively improves the dissolution rate of active ingredients such as flavonoids, alkaloids and polyphenols in chrysanthemum, lotus leaf and kudzu root, avoids the destruction of active ingredients by high temperature, and lays the raw material foundation for subsequent fermentation and fat reduction effects; 90℃ short-time enzyme inactivation sterilization removes impurities and bacteria in the raw materials and reduces the loss of plant active ingredients.

[0029] (2) In the technical solution of this invention, the fermentation strain is composed of compound *Lactobacillus plantarum* LP-Onlly and *Akermansia muciniphila* AKK016 and their ratio is optimized. At the same time, a 37℃ micro-anaerobic fermentation process is combined with low-speed intermittent stirring during the fermentation process, which better ensures the growth and metabolic environment of probiotics, so that the number of viable bacteria in the fermentation system is not less than 1×10⁻⁶. 9 CFU / g; In addition, the addition of EGCG not only enhances the antioxidant properties of the product, but also protects the activity of probiotics, effectively reducing the inhibitory effect of plant ingredients on probiotics.

[0030] (3) In the technical solution of the present invention, the plant active ingredients of chrysanthemum powder, lotus leaf powder and kudzu root powder and Lactobacillus plantarum LP-Onlly and Akkermansia muciniphila AKK016 are realized to achieve the synergistic fat reduction effect. The plant active ingredients directly play the role of reducing fat and inhibiting fat accumulation. The two strains improve lipid metabolism by regulating intestinal flora, enhancing intestinal barrier and promoting fat decomposition. The combination of the two greatly improves the fat reduction effect of the product. Moreover, the pH of the product after fermentation is weakly acidic, which improves the stability and bioavailability of plant active ingredients.

[0031] (4) The preparation process designed in the technical solution of the present invention is simple and easy to industrialize. It can be prepared into two dosage forms: liquid beverage or compound powder. The prepared product has a high number of live bacteria, good stability, and is convenient to eat. It is suitable for the weight loss needs of the general population and has both the safety and functionality of food. Detailed Implementation

[0032] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0033] The specific parameters of the raw materials used in this invention are as follows: Chrysanthemum powder, lotus leaf powder, kudzu root powder, and EGCG are all supplied by Hunan Jinong Biological Resources Co., Ltd., with product codes as follows: Hangbai Juyuan Chrysanthemum Powder (KL-CTEP012), Lotus Leaf Powder (KL-LLPE102), Kudzu Root Powder (KL-GGCE002), and EGCG (KL-TNEE003). Lactobacillus plantarum LP-Onlly (Pronova® Lactobacillus plantarum powder, viable count ≥1×10⁻⁶) 11 CFU / g) and Akkermansia myxophilus AKK016 (Pronova® Akkermansia myxophilus, viable count ≥1×10⁻⁶). 10CFU / g) were all provided by Shanghai Jiaotong University Angli Co., Ltd. In the following examples and comparative examples, the stirring and dispersion in step S3 were uniform, and the stirring rate was 120 rpm if not specified. In step S5, the low-temperature concentration was carried out using vacuum concentration with a vacuum degree of -0.08 MPa, a concentration temperature of 40°C, and a stirring rate of 50 rpm.

[0034] Example 1 A method for preparing a chrysanthemum powder composition with fat-reducing effects includes the following steps: S1: Take 70 parts by weight of chrysanthemum powder, 10 parts by weight of lotus leaf powder and 1 part by weight of kudzu root powder, and pulverize them separately at 0℃. Pass the chrysanthemum and lotus leaf powder through a 40-mesh sieve and the kudzu root powder through a 60-mesh sieve. After mixing, a mixed powder is obtained. S2: Add 8 times the volume of purified water to the mixed powder, soak at 50℃ for 30 min, then perform ultrasonic-assisted solubilization pretreatment (ultrasonic power of 200W, temperature of 45℃, time of 20 min), then heat at 90℃ for 10 min to inactivate enzymes and sterilize, and cool to 37±1℃ to obtain plant-based liquid. S3: Based on the dry weight of the raw materials, add 1‰ EGCG to the plant substrate liquid, then inoculate with fermentation bacteria (0.5‰ Lactobacillus plantarum LP-Onlly and 0.3‰ Akkermansia muciniphila), with a total inoculation amount of 0.8‰. Stir and disperse evenly to obtain the fermentation system. S4: Place the fermentation system in a micro-anaerobic environment with a carbon dioxide volume fraction of 5% and statically ferment at 37℃ for 24 hours, stirring at a low speed of 30 r / min for 5 minutes every 2 hours, until the system pH is 3.6 and the viable cell count is 1.1 × 10⁻⁶. 9 CFU / g was used to obtain the fermentation broth; S5: The fermentation broth was refrigerated at 4℃ for 2 hours, centrifuged at 3000 r / min for 15 minutes, filtered through a 200-mesh food-grade stainless steel filter, concentrated at low temperature to a solid content of 15%, and then freeze-dried (pre-frozen at -80℃ for 14 hours, then freeze-dried at -45℃ for 24 hours) and passed through an 80-mesh sieve to obtain a chrysanthemum powder composition with fat-reducing effect.

[0035] Example 2 A method for preparing a chrysanthemum powder composition with fat-reducing effects includes the following steps: S1: Take 80 parts by weight of chrysanthemum powder, 15 parts by weight of lotus leaf powder and 5 parts by weight of kudzu root powder, and pulverize them separately at a low temperature of 3℃. Pass the chrysanthemum and lotus leaf powder through a 40-mesh sieve and the kudzu root powder through a 60-mesh sieve. After mixing, a mixed powder is obtained. S2: Add 9 times the volume of purified water to the mixed powder, soak at 52℃ for 30 min, then perform ultrasonic-assisted solubilization pretreatment (ultrasonic power of 250W, temperature of 48℃, time of 20 min), then heat at 90℃ for 10 min to inactivate enzymes and sterilize, and cool to 37±1℃ to obtain plant-based liquid. S3: Based on the dry weight of the raw materials, add 2‰ EGCG to the plant substrate liquid, then inoculate with fermentation bacteria (0.6‰ Lactobacillus plantarum LP-Onlly and 0.4‰ Akkermansia muciniphila AKK016), with a total inoculation amount of 1.0‰. Stir and disperse evenly to obtain the fermentation system. S4: Place the fermentation system in a micro-anaerobic environment with a carbon dioxide volume fraction of 6.5% and statically ferment at 37℃ for 30 hours, stirring at a low speed of 30 r / min for 5 minutes every 2 hours, until the system pH is 3.8 and the viable cell count is 1.5 × 10⁻⁶. 9 CFU / g was used to obtain the fermentation broth; S5: The fermentation broth was refrigerated at 4℃ for 2 hours, centrifuged at 3500 r / min for 12 minutes, filtered through a 200-mesh food-grade stainless steel filter, concentrated at low temperature to a solid content of 18%, and then freeze-dried (pre-frozen at -80℃ for 14 hours, then freeze-dried at -45℃ for 24 hours) and passed through an 80-mesh sieve to obtain a chrysanthemum powder composition with fat-reducing effect.

[0036] Example 3 A method for preparing a chrysanthemum powder composition with fat-reducing effects includes the following steps: S1: Take 90 parts by weight of chrysanthemum powder, 20 parts by weight of lotus leaf powder and 10 parts by weight of kudzu root powder, and pulverize them separately at a low temperature of 5℃. The chrysanthemum and lotus leaf powder are passed through a 40-mesh sieve, and the kudzu root powder is passed through a 60-mesh sieve. After mixing, a mixed powder is obtained. S2: Add 10 times the amount of purified water to the mixed powder, soak at 55℃ for 30 min, then perform ultrasonic-assisted solubilization pretreatment (ultrasonic power of 300W, temperature of 50℃, time of 20 min), then heat at 90℃ for 10 min to inactivate enzymes and sterilize, and cool to 37±1℃ to obtain plant base liquid; S3: Based on the dry weight of the raw materials, add 3‰ EGCG to the plant substrate liquid, then inoculate with fermentation bacteria (0.7‰ Lactobacillus plantarum LP-Onlly and 0.5‰ Akkermansia muciniphila), with a total inoculation amount of 1.2‰. Stir and disperse evenly to obtain the fermentation system. S4: The fermentation system was placed in a micro-anaerobic environment with a carbon dioxide volume fraction of 8% and statically fermented at 37℃ for 36 hours, with low-speed stirring at 30 r / min for 5 minutes every 2 hours, until the system pH reached 4.0 and the viable cell count reached 1.8 × 10⁻⁶. 9 CFU / g was used to obtain the fermentation broth; S5: The fermentation broth was refrigerated at 4℃ for 2 hours, centrifuged at 4000 r / min for 10 minutes, filtered through a 200-mesh food-grade stainless steel filter, concentrated at low temperature to a solid content of 20%, and then freeze-dried (pre-frozen at -80℃ for 14 hours, then freeze-dried at -45℃ for 24 hours) and passed through an 80-mesh sieve to obtain a chrysanthemum powder composition with fat-reducing effect.

[0037] Comparative Example 1 The difference between this comparative example and Example 3 is that EGCG was not added in step S3, while the remaining steps, raw materials and parameters are the same as in Example 3.

[0038] Comparative Example 2 The difference between this comparative example and Example 3 is that in step S3, only *Lactobacillus plantarum* LP-Onlly 1.2‰ (without the synergistic effect of *Ackermania AKK016*) was inoculated. The remaining steps, raw materials and parameters are consistent with those in Example 3.

[0039] Comparative Example 3 The difference between this comparative example and Example 3 is that open aerobic fermentation (i.e., a carbon dioxide-free environment) is used in step S4, while the remaining steps, raw materials and parameters are consistent with those in Example 3.

[0040] Comparative Example 4 The difference between this comparative example and Example 3 is that the fermentation process in step S4 was not intermittently stirred at low speed, and the entire process was static micro-anaerobic fermentation. The remaining steps, raw materials and parameters were consistent with those in Example 3.

[0041] The fat-reducing efficacy of the chrysanthemum pollen compositions prepared in Examples 1-3 and Comparative Examples 1-4 was tested in C57BL / 6J high-fat model mice. The mice were administered the test substances continuously for 36 days, and indicators such as body weight, Lee's index, abdominal fat, serum lipids, liver lipids, and lipid metabolism-related enzyme activity were measured. The specific test methods are as follows: SPF-grade male C57BL / 6J mice (18-20g) were selected. The high-fat model group was fed a high-fat purified diet with 60% fat as the energy source for 8 weeks, with a body weight 20% higher than the negative control group as the model establishment standard. After successful modeling, mice were divided into a negative control group, a model control group, Example 1-3 groups, and Comparative Example 1-4 groups, with 8 mice in each group. The test substance was administered by gavage at a dose of 0.2g / kg per day, while the negative / model control groups were administered an equal volume of pure water by gavage. This treatment continued for 36 days. Body weight, body length, and food intake were measured weekly, and Lee's index was calculated: Lee's index = body weight (g)^(1 / 3) × 10 / body length (cm). After a 12-hour fast following the last gavage, liver / abdominal fat weight and organ indices were measured. Serum TC, TG, LDL-C, HDL-C, and liver TC, TG, NEFA, FAS, and HL were detected using reagent kits. SPSS 30.0 software was used for analysis. Quantitative data were presented as follows: The results indicate that one-way ANOVA was used for comparisons between groups, with P<0.05 indicating a significant difference and P<0.01 indicating a highly significant difference.

[0042] The test results are shown in Tables 1, 2 and 3.

[0043] Table 1. Test data of the baseline control group

[0044] Table 2 Test Results of Examples 1-3

[0045] Table 3 Test results of Comparative Examples 1-4

[0046] As can be seen from the data in Tables 1, 2 and 3, the product efficacy obtained in Examples 1-3 is better than that in Comparative Examples 1-4.

[0047] As can be seen from the results of Comparative Example 1, the lack of EGCG leads to the loss of antioxidant protection for probiotics. They are inhibited by polyphenolic components in plant raw materials, resulting in a significant decrease in the number of viable bacteria in the fermentation system and a weakening of metabolic activity. In addition, the loss of EGCG's own fat-reducing effect means that the synergistic fat-reducing effect is lost due to the reliance on the single effect of plant components and probiotics.

[0048] As can be seen from the results of Comparative Example 2, fermentation using only a single Lactobacillus plantarum LP-Onlly, without the metabolic symbiosis of Akkermansia muciniphila AKK016, cannot achieve the effects of enhancing the intestinal barrier and targeting the reduction of body fat percentage. Furthermore, the nutrient utilization efficiency in the fermentation system is reduced, and the number of viable bacteria is lower than that of the two-strain combination. Therefore, it is significantly inferior to Example 3 in terms of body fat percentage and blood lipid regulation indicators.

[0049] As shown in Comparative Example 3, *Ackermania myxophilus* AKK016, one of the two bacterial strains, is a strict anaerobic strain. An aerobic environment directly hinders its metabolism and significantly reduces its survival rate. The fermentation system cannot form a stable acidic environment, which inhibits the growth of other microorganisms and affects the conversion efficiency of plant active ingredients. The micro-anaerobic environment in this example better suits the metabolic characteristics of the two bacterial strains, ensuring continuous and efficient metabolism of the probiotics. Fermentation products such as organic acids and bioactive peptides work synergistically with plant active ingredients, thus significantly outperforming Comparative Example 3 in terms of lipid regulation indicators such as decreased serum TC / LDL-C and increased HDL-C.

[0050] As shown in Comparative Example 4, static fermentation easily leads to localized accumulation of nutrients and localized inactivation of probiotics, resulting in significant differences in metabolic efficiency across different regions of the fermentation system. In some regions, probiotics cannot fully metabolize due to insufficient nutrients, while in others, excessively high component concentrations cause inhibition. The intermittent low-speed stirring in this example maintains a stable micro-anaerobic environment while ensuring sufficient contact between nutrients and probiotics, guaranteeing uniform distribution of fermentation products and complete conversion of active ingredients. Therefore, it is significantly superior to Comparative Example 4 in terms of indicators such as reduced Lee's index and inhibition of liver FAS activity.

[0051] In the description of this specification, the references to terms such as "an embodiment," "example," "specific example," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0052] The above description is merely an example and illustration of the present invention. Those skilled in the art can make various modifications or additions to the specific embodiments described or use similar methods to replace them, as long as they do not deviate from the invention or exceed the scope defined by the present invention, they should all fall within the protection scope of the present invention.

Claims

1. A method for preparing a chrysanthemum powder composition with fat-reducing effects, characterized in that, Includes the following steps: S1: Take chrysanthemum powder, lotus leaf powder and kudzu root powder, sift them separately, and mix them to obtain a mixed powder; S2: Add the mixed powder to purified water, soak at 50-55℃, then perform ultrasonic solubilization pretreatment, cool after enzyme inactivation sterilization to obtain plant-based liquid; S3: Based on the dry weight of the raw materials, add epigallocatechin gallate to the plant substrate liquid, then inoculate with fermentation bacteria. The total inoculation amount is 0.5-1.5‰. Stir and disperse evenly to obtain the fermentation system. S4: Allow the fermentation system to undergo static micro-anaerobic fermentation for 24-36 hours, stirring for 5-8 minutes every 1.5-2 hours, until the system pH reaches 3.6-4.0 and the viable cell count is ≥1×10⁻⁶. 9 CFU / g was used to obtain the fermentation broth; S5: After the fermentation liquid is refrigerated and allowed to stand for 1-2 hours, it is centrifuged, filtered, and concentrated at low temperature until the solid content is 15%-20%. Then, it is freeze-dried and sieved to obtain a chrysanthemum powder composition with fat-reducing effect.

2. The method for preparing a chrysanthemum powder composition with fat-reducing effect according to claim 1, characterized in that, In step S1, the mass ratio of chrysanthemum powder, lotus leaf powder, and kudzu root powder is 70-90:10-20:1-10.

3. The method for preparing a chrysanthemum powder composition with fat-reducing effect according to claim 1, characterized in that, In step S1, the temperature of the material is 0-5℃ during sieving.

4. The method for preparing a chrysanthemum powder composition with fat-reducing effect according to claim 1, characterized in that, In step S1, the sieving process involves passing chrysanthemum powder and lotus leaf powder through a 40-mesh sieve, and kudzu root powder through a 60-mesh sieve.

5. The method for preparing a chrysanthemum powder composition with fat-reducing effect according to claim 1, characterized in that, In step S2, the ultrasonic-assisted dissolution pretreatment has an ultrasonic power of 200-300W, a temperature of 45-50℃, and a time of 20min.

6. The method for preparing a chrysanthemum powder composition with fat-reducing effect according to claim 1, characterized in that, In step S3, the fermentation strain composition based on the dry weight of the raw materials is: Lactobacillus plantarum LP-Onlly 0.3-1.0‰ and Akkermansia AKK016 0.2-0.5‰.

7. The method for preparing a chrysanthemum powder composition with fat-reducing effect according to claim 1, characterized in that, In step S4, the micro-anaerobic fermentation environment is an anaerobic culture environment with a CO2 volume fraction of 5%-8%.

8. The method for preparing a chrysanthemum powder composition with fat-reducing effect according to claim 1, characterized in that, In step S5, the freeze-drying process involves pre-freezing at -80°C for 14 hours, followed by freeze-drying at -45°C for 24 hours.

9. A method for preparing a chrysanthemum powder composition with a fat-reducing effect according to claim 1, characterized in that, The centrifugal filtration speed is 3000-4000 r / min, the time is 10-15 min, and the filtration uses a 200-mesh food-grade stainless steel filter screen.

10. A chrysanthemum powder composition with fat-reducing effect prepared by the preparation method according to any one of claims 1-9.

Images