Sucrose-free yogurt that can be stored at room temperature and its preparation method

Abstract

The present invention relates to a shelf-stable sucrose-free yogurt, its preparation method, and applications. The yogurt of the present invention is prepared by adding a stabilizer, a sweetener, inulin or xylooligosaccharide, whey protein, and honeysuckle extract to raw milk, and then fermenting the mixture with Streptococcus thermophilus, Lactobacillus bulgaricus, and Bifidobacterium longum BBMN68 as fermentation strains. The honeysuckle extract is an aqueous extract obtained by vacuum freeze-drying honeysuckle, crushing it, and infusing it with water. The yogurt of the present invention has the effects of regulating body weight, body fat percentage, and intestinal flora, and can therefore aid in weight loss.

Description

[Technical Field]

[0001] The present invention relates to the technical field of ambient yogurt, and in particular to shelf-stable sucrose-free yogurt and a method for preparing same. [Background technology]

[0002] Obesity, a disease caused by nutritional and metabolic imbalances, refers to a physiological condition in which the body has a relatively high fat content or is overweight. Survey results indicate that at least one-fifth of the world's population weighs more than the ideal weight level. Dieting is the act of reducing excess fat and weight. Weight loss can be achieved through a variety of methods, including lifestyle adjustments (diet, yoga, exercise, fitness, etc.), diet pills and diet foods, and surgical treatment. As research progresses, many dieters have discovered that dietary adjustments, appropriate aerobic exercise, and a disciplined lifestyle can help increase energy consumption and fat metabolism. Clinical studies have shown that combining scientific and rational nutritional therapy with exercise intervention remains the most effective and safe basic treatment to date.

[0003] Honeysuckle, the dried flower buds or first blooming flowers of honeysuckle (Lonicera japonica), a plant in the Caprifoliaceae family, is a commonly used traditional Chinese medicine with the same origins as medicines and foods, possessing various pharmacological effects, including anti-inflammatory, antiviral, and antioxidant properties. Research has shown that chlorogenic acid (CHA) is the organic acid component with the highest content of the main active ingredient in honeysuckle. Chlorogenic acid, or 3-caffeoylquinic acid, is a secondary metabolite produced by aerobic respiration in the plant. It has pharmacological activities, including antiviral, antihypertensive, hypolipidemic, hepatoprotective, cholestasis promotion, and free radical scavenging, making it a hot spot in the fields of food and pharmaceutical research. Because honeysuckle is rich in CHA, it is widely used in foods to demonstrate its beneficial effects.

[0004] Patent CN101595918B discloses a yogurt product containing polydextrose, inulin, and a special combination of fermented bacteria strains. This product is low in calories and can control energy intake, thereby playing a positive role in weight control. Patent CN102870882B discloses that this product achieves the effect of lowering blood sugar levels by adding various traditional Chinese medicine ingredients that are claimed to have a weight-loss effect, and is also claimed to have a weight-regulating effect. Patent CN112273565A discloses a process for preparing a liquid drink containing honeysuckle and chrysanthemum ingredients, in which honeysuckle is added to the drink by mixing and stirring, thereby giving the drink a cooling and detoxifying effect. Patent CN113142303A discloses that special lactic acid bacteria and mulberry leaf extract gel particles are added to adjust the intestinal flora, and intragastric experiments using high-fat diet mice verify that this product has the effect of preventing the onset of obesity in various ways, such as regulating blood lipid and cholesterol levels.

[0005] Currently, there are not many ambient temperature yogurt products on the market that claim to have weight loss effects, and those that do have weight loss effects still have many issues that need to be optimized or improved. The formulation design of most anti-fat yogurt products at this stage is complicated and tedious. Some yogurts contain too many types of fermenting bacteria strains, which can result in long fermentation times and make it difficult to control the fermentation conditions. Some yogurts contain multiple functional ingredients to improve the product's weight control ability, but this actually affects the product's flavor, texture, and shelf life. Summary of the Invention [Problem to be solved by the invention]

[0006] Yogurt is a type of fermented dairy product with high nutritional value, which is in high demand from the market and consumers. Currently, consumer demand for weight loss and dieting has led to the development of various yogurts, and sugar-free and low-fat yogurt products are one of the hotspots in development.

[0007] An object of the present invention is to provide a sucrose-free yogurt fermented milk product that can be stored at room temperature and has a weight control (diet) effect. [Means for solving the problem]

[0008] To achieve the object of the present invention, in a first aspect, the present invention provides a method for preparing shelf-stable sucrose-free yogurt, comprising: (2-10 g):(1-6 g):(1×10) honeysuckle extract, a prebiotic, and a probiotic, Bifidobacterium longum BBMN68, per 100 g of raw materials for the shelf-stable sucrose-free yogurt. 7~8 CFU) and the prebiotic is inulin or xylooligosaccharides.

[0009] The shelf-stable sucrose-free yogurt obtained by the present invention does not contain sucrose, thereby further reducing the calorie content of the yogurt. This makes the yogurt suitable for people who need to reduce fat intake. The shelf-stable sucrose-free yogurt without added sucrose obtained by the present invention has good flavor and stability, and the sucrose-free yogurt of the present invention satisfies stability requirements without placing an excessive weight burden on consumers.

[0010] The addition of chlorogenic acid makes the fermentation system of yogurt unstable, resulting in a short shelf life and prone to syneresis of water in the finished yogurt obtained by fermentation. The present invention significantly improves the stability of the fermentation system by adding prebiotics.

[0011] Both inulin and xylooligosaccharides are currently commercially available and widely used prebiotics, and are mixtures containing oligosaccharides or polysaccharides with different degrees of polymerization. The beneficial effects of prebiotics on the human body, such as regulating the intestinal microflora, strengthening the immune system, and controlling body weight, have been confirmed by numerous literature studies and human experiments.

[0012] It has been confirmed through experiments of the present invention that after inulin and xylooligosaccharides are added, the stability of the fermented milk system is improved, and the production of short-chain fatty acids during the fermentation process of the product is effectively increased, thereby enhancing the efficacy of yogurt in fat reduction and weight control.

[0013] In the prior art, functional yogurt products that promote intestinal health have been most widely developed, but functional yogurt products related to weight control still have a large market space. The yogurt prepared using honeysuckle in the present invention has a refreshing aroma, and its main component, chlorogenic acid, has a certain effect on weight control.

[0014] In a preferred embodiment, the method for extracting honeysuckle extract of the present invention comprises pre-freezing honeysuckle, freeze-drying the honeysuckle to obtain honeysuckle powder, dissolving the honeysuckle powder in sterile water at a solid-liquid ratio of 20:1 to 30:1 g / L in an insulated tank at 65 to 75°C, infusing for 10 to 20 minutes, and filtering to obtain honeysuckle extract.

[0015] In a preferred embodiment, after the fermentation of the shelf-stable sucrose-free yogurt is completed, the ingredients are preheated to 50-55°C and maintained for 5-10 minutes before sterilization and filling, thereby increasing the chlorogenic acid content of the final shelf-stable sucrose-free yogurt.

[0016] Specifically, in the preparation method of the present invention, the shelf-stable sucrose-free yogurt is prepared by adding a stabilizer, a sweetener, prebiotics, whey protein, and honeysuckle extract to raw milk, and fermenting the mixture with Streptococcus thermophilus, Lactobacillus bulgaricus, and Bifidobacterium longum BBMN68 as fermentation strains, wherein the stabilizer is highly esterified pectin, and the sweetener is erythritol or xylitol.

[0017] In a preferred embodiment, the ingredients of the shelf-stable sucrose-free yogurt, based on the percentage of the actual total weight, include 80-100 parts by weight of raw milk, 2-10 parts by weight of honeysuckle extract, 0.01-10 parts by weight of sweetener, 0.5-2 parts by weight of whey protein powder, 0.1-0.5 parts by weight of highly esterified pectin, 1-6 parts by weight of prebiotics, and 1×10 effective viable cell count. 7~10 CFU / 100g of Lactobacillus bulgaricus, effective viable cell count 1 x 10 7~10 CFU / 100g of Streptococcus thermophilus and effective viable bacteria count 1 x 10 7~8 It contains CFU / 100g of Bifidobacterium longum BBMN68, which has the deposit number CGMCC NO. 2265 and is disclosed in Chinese Patent CN101649303B.

[0018] In one specific embodiment, the method for preparing shelf-stable sucrose-free yogurt according to the present invention comprises: Step (1) of filtering raw milk through a reverse osmosis membrane to obtain a retentate and a permeate; The retentate described in step (1) is preheated to 45-55°C, and the sweetener, whey protein powder, prebiotics, and honeysuckle extract are added, stirred uniformly, homogenized, sterilized, and cooled. (2) introducing Streptococcus thermophilus, Lactobacillus bulgaricus, and the probiotic Bifidobacterium longum BBMN68, fermenting for 4 to 8 hours, fermenting until the pH reaches 4.25 to 4.35, demulsifying, and cooling to obtain a first material; Step (3) of heating the permeate separated in step (1) to 50-55°C, adding a stabilizer, uniformly stirring, homogenizing, sterilizing, and cooling to obtain a second material; and step (4) statically mixing the first material obtained in step (2) and the second material obtained in step (3), preheating to 50-60°C and maintaining the temperature for 2-5 minutes, then increasing the temperature to 65-80°C and sterilizing for 15-60 seconds, cooling to 20-25°C, aseptically homogenizing at 20-30 bar, and aseptically filling to obtain room temperature shelf-stable sucrose-free yogurt.

[0019] In one specific embodiment, in step (2) of the preparation method according to the present invention, the homogenization conditions before fermentation are 55-65°C and 150-160 bar, the sterilization conditions are 120-130°C and 5-10 s, the cooling temperature is 40-42°C, and the temperature after demulsification is 20-25°C.

[0020] In one specific embodiment, in step (3) of the preparation method according to the present invention, after adding a stabilizer and stirring uniformly, the homogenization conditions are 55-65°C and 150-160 bar, the sterilization conditions are 110-120°C and 15 s, and the cooling temperature is 20-25°C.

[0021] In one specific embodiment of the present invention, a method for preparing shelf-stable sucrose-free yogurt includes: The screened and washed honeysuckle was pre-frozen in a refrigerator at -80°C, and then dried for 24 hours under the conditions of a cold trap temperature of -50°C, a partition heating temperature of 20°C, and a vacuum of 0.12 mbar to obtain honeysuckle powder. The freeze-dried powder was pulverized and dissolved in sterilized water at a solid-liquid ratio of 30:1 g / L. The powder was then infused in a heated tank at 65-75°C for 20 minutes. Step (1) of filtering to obtain a honeysuckle water extract; (2) filtering the raw milk through a reverse osmosis membrane to obtain a retentate and a permeate; Step (3) of preheating the retentate described in step (2) to 55 ° C, adding sugar replacer, whey protein powder, inulin or xylooligosaccharide, honeysuckle extract, stirring uniformly, and homogenizing under conditions of 60 ° C and 150 bar; The homogenized material is sterilized at 121°C for 6 seconds, then cooled to 40°C, and yogurt starter (Streptococcus thermophilus and Lactobacillus bulgaricus) and probiotic (Bifidobacterium longum BBMN68) are added and fermented for 6 hours until the pH reaches 4.35 and the acidity reaches 65°C or more. After that, the material is demulsified and cooled to 20°C to obtain a first material. The fermentation temperature is 40-45°C, and the fermentation time is 4-8 hours until the pH reaches 4.35 and the acidity reaches 65°C or more. After that, the material is demulsified, thereby promoting the release of chlorogenic acid from the honeysuckle infusion. (4) Step (5) of heating the permeate separated in step (3) to 55°C, adding a stabilizer, stirring uniformly, and homogenizing under conditions of 60°C and 150 bar. After homogenization is completed, sterilizing under conditions of 110°C for 15 seconds, and cooling to 25°C to obtain a second material; Next, the first material obtained in step (4) and the second material obtained in step (5) are statically mixed, preheated to 55°C and maintained for 5 minutes, the preheating process is favorable for further release of chlorogenic acid, and then heated to 65°C and sterilized for 60 seconds (step (6)); and step (7) aseptically homogenizing the mixed and sterilized material in step (6) at 25°C and 20 bar, and aseptically filling the homogenized material to obtain shelf-stable sucrose-free yogurt.

[0022] The recommended intake of the shelf-stable sucrose-free yogurt of the present invention is 100 g twice a day.

[0023] In a second aspect, the present invention seeks to protect warm zero-sucrose yogurt prepared by the above preparation method.

[0024] As will be appreciated by those skilled in the art, the present invention also seeks to protect the application of said shelf-stable sucrose-free yogurt in the manufacture of health foods for fat reduction and body shaping. [Effects of the Invention]

[0025] The present invention relates to a shelf-stable, sucrose-free fermented yogurt milk product with weight control (diet) effects. This fermented milk product contains inactivated Bifidobacterium longum BBMN68, inulin, and honeysuckle extract. The present application optimizes the honeysuckle extract processing to increase the chlorogenic acid content of the final product and promote its fat-reducing effect. Next, the product is developed by adding a probiotic (Bifidobacterium longum BBMN68) to the starter to participate in fermentation, thereby promoting the release of chlorogenic acid from the honeysuckle extract and improving the functionality of the final product. At the same time, the product also contains an inulin ingredient to ensure the product's flavor and shelf-life stability, helping to achieve its weight-loss effect. Compared to other shelf-stable yogurt formulations, the present invention's shelf-life regulating effects on factors such as body weight and body fat percentage in rats were verified through animal experiments.

[0026] More specifically, the beneficial effects of the present invention are one or more of the following: (1) The present invention solves the problem of low chlorogenic acid release in honeysuckle extract. Compared with existing honeysuckle extract processing methods, the present invention combines vacuum freeze-drying and leaching processes to increase the chlorogenic acid content of honeysuckle extract. The product of the present invention includes a preheating process before packaging, thereby increasing the chlorogenic acid content of the final product, shelf-stable sucrose-free yogurt. Chlorogenic acid is the organic acid component with the highest content in honeysuckle extract. It inhibits fat absorption, promotes cholesterol excretion, and has a diet effect.

[0027] (2) The product obtained by this invention is prepared by adding probiotics (Bifidobacterium longum BBMN68) to the yogurt starter to participate in the fermentation, which promotes the release of chlorogenic acid from the honeysuckle extract, which helps increase the chlorogenic acid content of the final product and enhances the diet effect of the product.

[0028] (3) The present invention solves the problem of instability in yogurt fermentation systems caused by the addition of chlorogenic acid. The product obtained by the present invention contains inulin, which ensures the stability of the product at room temperature throughout its shelf life and simplifies the product formulation. At the same time, inulin, as a type of prebiotic, helps promote the metabolism of human intestinal flora to produce short-chain fatty acids, which contribute to achieving weight control effects in the intestine.

[0029] In order to more clearly explain the technical solutions of the present invention or the prior art, the following briefly describes the drawings required in the description of the embodiments or prior art. Obviously, the drawings described below are only some embodiments of the present invention, and those skilled in the art can also derive other drawings based on these drawings without any creative efforts. [Brief explanation of the drawings]

[0030] [Figure 1] Figure 1 shows the chlorogenic acid content of honeysuckle extract of the present invention before and after optimizing the extraction process, where a indicates no difference from the direct infusion group, b indicates a difference from the direct infusion group (p<0.05), and c indicates a difference from both the direct infusion group and the vacuum freeze-dried group (p<0.05). [Figure 2] FIG. 1 shows the effect of the heat treatment of the present invention on the chlorogenic acid content of the product, where a shows that there is a difference (p<0.05) from the chlorogenic acid content of the infusion before heat treatment. [Figure 3] 1 shows the effect of varying the addition of the probiotics of the present invention on the chlorogenic acid content of the product and the effect of setting the process conditions of the examples on the chlorogenic acid content of the product. Different lowercase letters bc in the figure indicate differences (p<0.05) between different groups, and groups marked with the same letter indicate no significant difference between these groups. [Figure 4]1 shows the results of detecting short-chain fatty acids in an example of the present invention and a comparative example, where a indicates no difference from Comparative Example 4, b indicates a difference from Comparative Example 4 (p<0.05), and c indicates a difference from both Comparative Example 4 and Example 4 (p<0.05). [Figure 5] 1 shows the effects of the present invention and comparative examples on rat weight gain. Different letters (abcde) in the figure indicate differences (p<0.05) between different groups, and groups marked with the same letter indicate no significant difference between these groups. [Figure 6] 1 shows the effects of the present invention and comparative examples on rat body fat percentage. In the figure, different letters ab indicate differences between different groups (p<0.05), and groups marked with the same letter indicate no significant difference between these groups. b indicates no significant difference from the model group, but a significant difference from the blank group (p<0.05). [Figure 7] FIG. 1 shows the results of detecting acetic acid in the feces of rats from an example of the present invention and a comparative example, where * indicates a significant difference from other groups (p<0.05). [Figure 8] FIG. 1 shows the results of detecting propionic acid in the feces of rats from an example of the present invention and a comparative example, where * indicates a significant difference from other groups (p<0.05). [Figure 9] FIG. 1 shows the results of detecting butyric acid in the feces of rats from an example of the present invention and a comparative example, where * indicates a significant difference from other groups (p<0.05). DETAILED DESCRIPTION OF THE INVENTION

[0031] In order to make the objectives, technical solutions and advantages of the present invention clearer, the technical solutions of the present invention are described clearly and comprehensively below, and obviously, the described embodiments are only a part of the embodiments of the present invention, but not all of the embodiments. Based on the embodiments of the present invention, all other embodiments that a person skilled in the art can obtain without any creative efforts fall within the protection scope of the present invention.

[0032] In one embodiment of the present invention, honeysuckle extract is prepared as follows: Select honeysuckle with full flower buds, emerald green color, and no mold, insect damage, scattered leaves, or broken branches. Surface impurities such as mud and sand are removed by washing with clean water. The honeysuckle is pre-frozen in a refrigerator at -80°C, then placed in a vacuum freeze-drying machine and dried for 24 hours under conditions of a cold trap temperature of -50°C, a partition heating temperature of 20°C, and a vacuum of 0.12 mbar. Honeysuckle powder is obtained by pulverizing the freeze-dried powder and adding sterilized water at a solid-liquid ratio of 20:1 to 30:1 g / L. The mixture is then infused in an incubator at 65-75°C for 20 minutes. The resulting mixture is then filtered to obtain a honeysuckle aqueous extract.

[0033] Lactobacillus bulgaricus and Streptococcus thermophilus are strains commonly purchased commercially.

[0034] Example 1 Preparation of shelf-stable sucrose-free yogurt The ingredients added to the fermented milk product (according to mass percentage) were honeysuckle extract 2%, erythritol 0.7%, xylitol 0.9%, whey protein powder 0.6%, highly esterified pectin 0.2%, inulin 3%, Lactobacillus bulgaricus 0.0001% (when the concentration of bacteria in the starter was 1 × 10 12 CFU / g), Thermophilus Streptococcus 0.0001% (concentration of bacteria in the starter is 1 × 10 12 CFU / g), Bifidobacterium longum BBMN68 0.02% (bacterial concentration 5 × 10 8 CFU / g, 1 x 10 9 The remaining portion was supplemented with raw milk.

[0035] The method for preparing this shelf-stable sucrose-free yogurt is specifically as follows: (1) The screened and washed honeysuckle is pre-frozen in a -80°C refrigerator and dried for 24 hours under conditions of a cold trap temperature of -50°C, a partition heating temperature of 20°C, and a vacuum of 0.12 mbar to prepare honeysuckle powder. The freeze-dried powder is pulverized and then added to sterilized water at a solid-liquid ratio of 30:1 g / L. The mixture is dissolved in an incubator at 75°C and infused for 20 minutes. The honeysuckle water extract is obtained by filtration. Filtration is typically performed at room temperature using a filter net or filter cloth to remove impurities.

[0036] (2) The raw milk is filtered through a reverse osmosis membrane to obtain a retentate and a permeate.

[0037] (3) The retentate described in (2) is preheated to 55°C, and the sugar replacer, whey protein powder, inulin, and honeysuckle extract are added. The mixture is stirred uniformly and homogenized at 60°C and 150 bar.

[0038] (4) The homogenized material was sterilized at 121°C for 6 seconds, then cooled to 40°C. Yogurt starter (Streptococcus thermophilus and Lactobacillus bulgaricus) and probiotic (Bifidobacterium longum BBMN68) were added and fermented for 6 hours until the pH reached 4.35 and the acidity reached 65°T. The material was then demulsified and cooled to 20°C to obtain the first material. This fermentation time and temperature setting contributed to the release of chlorogenic acid from the honeysuckle infusion.

[0039] (5) The permeate separated in (2) is heated to 55°C, a stabilizer is added, the mixture is stirred uniformly, and the mixture is homogenized at 60°C and 150 bar. After homogenization is complete, the mixture is sterilized at 110°C for 15 seconds and cooled to 25°C to obtain the second material.

[0040] (6) Next, the first material obtained in (4) and the second material obtained in (5) are statically mixed, preheated to 55°C and maintained for 5 minutes, as the preheating process favors further release of chlorogenic acid, and then heated to 65°C and sterilized for 60 seconds.

[0041] (7) The ingredients mixed and sterilized in (6) are aseptically homogenized at 25°C and 20 bar, and then aseptically filled to obtain sucrose-free yogurt that can be stored at room temperature.

[0042] Example 2 Preparation of shelf-stable sucrose-free yogurt The ingredients added to the fermented milk product (according to mass percentage) were honeysuckle extract 5%, erythritol 0.7%, xylitol 0.9%, whey protein powder 0.6%, highly esterified pectin 0.2%, inulin 3%, Lactobacillus bulgaricus 0.0001% (the concentration of bacteria in the starter was 1 × 10 12 CFU / g), Thermophilus Streptococcus 0.0001% (concentration of bacteria in the starter is 1 × 10 12 CFU / g), Bifidobacterium longum BBMN68 0.2% (bacterial concentration 5 × 10 8 CFU / g, 1 x 10 10 The remaining portion was supplemented with raw milk.

[0043] The method for preparing this shelf-stable sucrose-free yogurt was the same as in Example 1.

[0044] Example 3 Preparation of shelf-stable sucrose-free yogurt The ingredients added to the fermented milk product (according to mass percentage) were honeysuckle extract 8%, erythritol 0.7%, xylitol 0.9%, whey protein powder 0.6%, highly esterified pectin 0.2%, inulin 3%, Lactobacillus bulgaricus 0.0001% (the concentration of bacteria in the starter was 1 × 10 12 CFU / g), Thermophilus Streptococcus 0.0001% (concentration of bacteria in the starter is 1 × 10 12 CFU / g), Bifidobacterium longum BBMN68 0.02% (bacterial concentration 5 × 10 8 CFU / g, 1 x 10 9 The remaining portion was supplemented with raw milk.

[0045] The method for preparing this shelf-stable sucrose-free yogurt was the same as in Example 1.

[0046] Example 4 Preparation of shelf-stable sucrose-free yogurt The ingredients added to the fermented milk product (according to mass percentage) were honeysuckle extract 5%, erythritol 0.7%, xylitol 0.9%, whey protein powder 0.6%, highly esterified pectin 0.2%, xylooligosaccharides 1.5%, Lactobacillus bulgaricus 0.0001% (the concentration of bacteria in the starter was 1 × 10 12 CFU / g), Thermophilus Streptococcus 0.0001% (concentration of bacteria in the starter is 1 × 10 12 CFU / g), Bifidobacterium longum BBMN68 0.02% (bacterial concentration 5 × 10 8 CFU / g, 1 x 10 9 The remaining portion was supplemented with raw milk.

[0047] The method for preparing this shelf-stable sucrose-free yogurt was the same as in Example 1.

[0048] Example 5 Preparation of shelf-stable sucrose-free yogurt The ingredients added to the fermented milk product (according to mass percentage) were honeysuckle extract 5%, erythritol 0.7%, xylitol 0.9%, whey protein powder 0.6%, highly esterified pectin 0.2%, inulin 3%, Lactobacillus bulgaricus 0.0001% (the concentration of bacteria in the starter was 1 × 10 12 CFU / g), Thermophilus Streptococcus 0.0001% (concentration of bacteria in the starter is 1 × 10 12 CFU / g), Bifidobacterium longum BBMN68 0.02% (bacterial concentration 5 × 10 8 CFU / g, 1 x 10 9 The remaining portion was supplemented with raw milk.

[0049] The method for preparing this shelf-stable sucrose-free yogurt was the same as in Example 1.

[0050] Comparative Example 1 Preparation of sucrose-free yogurt that can be stored at room temperature In this comparative example, unlike Example 5, no honeysuckle extract was added, and the dosages and preparation methods of the other ingredients were the same as those in Example 5.

[0051] Comparative Example 2 Preparation of room temperature sucrose-free yogurt In this comparative example, 15% honeysuckle extract was added to the composition of Example 5, and the dosages and preparation methods of the other ingredients were the same as those of Example 5.

[0052] Comparative Example 3 Preparation of room temperature sucrose-free yogurt In this comparative example, Bifidobacterium longum was not added, and the dosages and preparation methods of the other ingredients were the same as in Example 5.

[0053] Comparative Example 4: Different Prebiotics In this comparative example, polydextrose was selected as the prebiotic in the raw material, and the dosage and preparation method of the other formulations were all the same as those in Example 5.

[0054] Comparative Example 5: Different Probiotics In this comparative example, Lactobacillus BB-12 (manufactured by Christian Hansen, Denmark) was selected as the probiotic raw material, and the dosage and preparation method of the other ingredients were all the same as in Example 5.

[0055] Comparative Example 6 Different fermentation conditions Compared with Example 5, the difference between this comparative example and Example 5 is that the fermentation conditions are 38°C.

[0056] Comparative Example 7 Different preheating treatments before filling Compared with Example 5, the difference between this comparative example and Example 5 is that in this comparative example, no preheating treatment is performed before filling.

[0057] Experimental Example 1: Analysis of product flavor and stability The shelf-stable sucrose-free yogurts prepared in Examples 1 to 5 and Comparative Examples 1 to 4 were tested for sensory suitability. The test method was as follows: 50 people were selected to conduct a blind test on each of the shelf-stable sucrose-free yogurts. The blind test index included characteristic flavor and texture (consistency), and was expressed numerically as follows: 1-2 represented very weak, 3-4 represented weak, 5-6 represented good, 7-8 represented strong, and 9-10 represented very strong. After the test was completed, a statistical analysis was performed on the test results, and the results are shown in Table 1:

[0058] [Table 1]

[0059] Conclusion: By comparing and analyzing the evaluation results of the Examples and Comparative Examples in Table 1, it can be seen that adding honeysuckle extract within the scope of the Examples has the effect of improving the characteristic flavor compared to Comparative Examples 1 and 2, but does not cause the problem of the characteristic flavor being too prominent and affecting the overall flavor of the product.

[0060] Compared with the Examples, analysis of the results of Comparative Examples 3 and 4 shows that the addition of probiotics, inulin, and xylooligosaccharides can improve the overall characteristic flavor and texture of the product. Analysis of the results of Comparative Example 2 shows that adjusting the amount of honeysuckle extract added also affects the texture of the product. Unlike the Comparative Examples, the honeysuckle room temperature zero sucrose yogurt prepared in the Examples of the present invention has better overall sensory suitability and flavor preference.

[0061] The room temperature zero sucrose yogurts prepared in Examples 1 to 5 and Comparative Examples 1 to 4 were analyzed by centrifugal water loss rate and shelf life stability tests at 25°C and 37°C, and the results are shown in Table 2.

[0062] [Table 2]

[0063] The results in Table 2 show that the room temperature shelf-stable sucrose-free yogurts prepared in Examples 1 to 5 had low centrifugal water loss after 15 minutes of centrifugation at 4000 rpm, the product systems were relatively stable, and no significant syneresis or thinning occurred during room temperature or warm storage observation.

[0064] In the preparation method of Comparative Example 1, no honeysuckle extract was added, which had a slight effect on the stability of the product. The honeysuckle extract added in Comparative Example 2 was not within the limit range of the examples, and the imbalance of the additives prevented the formation of a good stable system. After about two months of incubation and storage, the product suffered significant water loss during centrifugation.

[0065] In Comparative Example 4, a probiotic program different from that of the present invention was used, and as a result, the protective ability of these probiotics in terms of stability was insufficient, resulting in serious syneresis in the product and the system collapsing after 2 months of storage at room temperature.

[0066] In Comparative Example 3, the probiotics of Example 5 were not used, and the other ingredients were the same, but the product was less smooth and thinned at the bottom after being stored at room temperature for two months.

[0067] This product combines honeysuckle extract and inulin to achieve a balanced flavor while maintaining room temperature product functionality, ensuring shelf life stability, and simplifying the product formulation.

[0068] Experimental Example 2: Detection and analysis of chlorogenic acid content The method for detecting the chlorogenic acid content of honeysuckle extract was designed with reference to relevant literature.

[0069] References: [1] Ding Min, Wang Liling, Qin Yuchuan et al. Research on the aqueous extraction process of chlorogenic acid in honeysuckle [J]. Zhejiang Forestry Science and Technology, 2022, 42(2):6. [2] Wang Yujie, Jie Weiguang, Guo Na et al. Simultaneous detection of chlorogenic acid and total flavonoids in honeysuckle by high-performance liquid chromatography and their extraction process [J]. Grains and Fats, 2022, 35(2):6. [3] Zou Rong, Yu Yuming, Chen Zexiong, Hu Kai, Ran Lie. Effect of drying methods on honeysuckle polyphenol components and their antioxidant activity [J]. Food Science, 2016, 37(5):6.

[0070] In this experiment, high-performance liquid chromatography was used for detection. First, a standard solution was prepared using a chlorogenic acid standard substance (standard product), and solutions with different concentration gradients were prepared by precisely absorbing the chlorogenic acid standard solution. A standard curve for chlorogenic acid content was created using high-performance liquid chromatography at a detection wavelength of 327 nm. Furthermore, using the same detection method, the chlorogenic acid content in the honeysuckle juice after infusion, the chlorogenic acid content after fermentation, and the chlorogenic acid content in the final product were detected and calculated.

[0071] (1) The effect of process optimization on the chlorogenic acid content of the product Currently, the methods for extracting honeysuckle include direct leaching, vacuum freeze-drying, and vacuum freeze-drying + leaching. The results in Figure 1 show that by comparing the two process methods of leaching and freeze-drying separation, the content leached by the combined process method is different from the other two groups. This shows that the present invention increases the content of chlorogenic acid in honeysuckle extract by combining the methods of vacuum freeze-drying and dissolution leaching.

[0072] When preparing room-temperature zero-sucrose yogurt, different heat sterilization methods have different effects on the chlorogenic acid content in the yogurt. Figure 2 shows that when honeysuckle infusion was added during mixing, the chlorogenic acid content of the honeysuckle extract after two different degrees of sterilization heat treatment increased significantly. Although there was no significant difference in the chlorogenic acid content of the samples after UHT heat treatment (sterilization treatment in step (4)) or UHT + pasteurization heat treatment (preheating and sterilization treatment in step (6)), the chlorogenic acid content of the samples in both groups increased significantly compared to the infusion before heat treatment, indicating that the processing of room-temperature zero-sucrose yogurt further promoted the release of the active ingredients from the honeysuckle extract.

[0073] From the results in Figure 3, when the fermentation conditions during fermentation were adjusted as in Comparative Example 6 or when the preheating treatment before sterilization was omitted as in Comparative Example 7, the chlorogenic acid content of the sample tended to decrease significantly, indicating the necessity of setting the process conditions in this example.

[0074] (2) Effect of probiotic addition on the chlorogenic acid content of products In the present invention, probiotics (Bifidobacterium longum BBMN68) are added to yogurt starter to participate in fermentation. A comparison of the results of Example 2, Comparative Examples 3, and 5 shows that the addition of Bifidobacterium longum BBMN68 can better promote the release of chlorogenic acid from honeysuckle extract (Figure 3). Comparing the results of Example 2 and Example 5 indicates that the number of probiotic bacteria is related to the chlorogenic acid content of the final product and may enhance the fat-reducing effect of the product.

[0075] Experimental Example 3: Detection of short-chain fatty acids in products This experimental example was based on the formulation of Example 5, and the short-chain fatty acid content of the prepared room-temperature zero-sucrose yogurt product was detected. The results are shown in Figure 4. For detection methods, please refer to the following literature: [1] Wu Xufang, Zhang Yangdong, Zheng Nan et al., Research Progress in the Composition, Physiological Function, and Detection Technology of Short-Chain Fatty Acids in Milk [J], Journal of Animal Nutrition, 2022, 34(04):2148-2155. [2] Fatty Acid Profile of Milk for Determining Reproductive Status in Lactating Holstein Friesian Cows, Hawar M. Zebari; S. Mark Rutter; Emma CL Bleach. Animal Reproduction Science, 2019.

[0076] The detection and analysis of short-chain fatty acids show that under the design of adding prebiotics to the formulation of the present invention, Examples 4 and 5 can effectively increase the production of short-chain fatty acids in the fermentation process of the products compared with Comparative Example 4, which can to some extent improve the effectiveness of the products in terms of fat reduction and weight control.

[0077] Experimental Example 4: Weight regulation and control effects Several SPF-grade Wistar rats, male, weighing 140-170g, were used in the experimental animal room, which consisted of a barrier system and an IVC cage box, with a temperature of 20-25°C and a relative humidity of 45%-65%. There were 10 rats per group. The blank group was fed a normal diet (intragastric administration of saline), the model group was fed a high-fat diet (intragastric administration of saline), and the test group was fed a high-fat diet (intragastric administration of fermented milk at 10g / kg per day). Weight changes in rats in each group were observed after 7 days of intragastric administration for a total of 12 weeks.

[0078] At the start of the experiment, animals in the model and blank groups were required to have a weight difference of more than 20% of their average weight. They were then divided into experimental groups containing different formulations. Test samples were orally administered, and their weight and food intake were measured twice weekly. At the end of the experiment, weight changes were calculated and body fat percentage was measured (using MRI-related equipment). For details on the measurement method, please refer to the following references: [1] Li Chenyang. Development and Application of a Non-Destructive Analysis Method for Mouse Body Composition [D]. Dalian University of Technology, 2018. [2] Choi B, Park S, Lee D, et al. Green coffee bean extract improves obesity by decreasing body fat in high-fat diet-induced obese mice [J]. Asian Pacific Journal of Tropical Medicine, 2016, 9(07):616-624.

[0079] The results of the evaluation of the weight-regulating effect of the product in this experimental example are shown in Figure 5. The results in Figure 5 show that the rats in each group gained weight to varying degrees, with significant differences between the model and control groups, indicating successful creation of an obesity model. Each example significantly reduced weight gain compared to the model and comparative groups. Although there was a difference between Example 5 and Example 4, both exhibited good weight-regulating effects. The difference between the comparative groups was generally not significant, and although there was a difference with the model group, the effect was weaker than that of the examples. There was no significant difference between the weight gain of Example 5 and the blank group, indicating a good weight-regulating effect. Example 5 achieved a reduction in weight gain compared to Comparative Example 1.

[0080] The results of this experimental example, in which the product's effect on rat body fat percentage was evaluated, are shown in Figure 6. The body fat percentage of rats in each group decreased to different degrees, with no significant difference between Examples 4 and 5 and the blank group, indicating the Examples' excellent effect on improving body fat. The Comparative Example showed a slight decrease compared to the model group, but the effect was general, and therefore the difference between the groups was not significant. Comparative Example 1 was intermediate between the indicators of the blank group and the model group, indicating that the zero-sucrose yogurt product itself has a certain effect on weight control. Example 5 also achieved a reduction in body fat percentage compared to Comparative Example 1.

[0081] Experimental Example 5 Distribution of intestinal flora in rats In this example experiment, the method for detecting and analyzing the rat intestinal microbiota involved intragastrically administering test samples to rats in different groups, selecting six rats from each group, and collecting all fresh feces within 5 hours at the end of the study. The fecal matter was then stored in a centrifuge tube with a lid, the total mass and fecal volume were recorded, and the tubes were stored at -80°C for measurement of the microbiota. The microbiota was detected using 16S rDNA sequencing. For detection methods, please refer to the following literature: [1]. WEI J, ZHAO Y, ZHOU C, et al. Dietary Polysaccharide from Enteromorpha clathrata Attenuates Obesity and Increases the Intestinal Abundance of Butyrate-Producing Bacterium, Eubacterium xylanophilum, in Mice Fed a High-Fat Diet[J]. Polymers (Basel), 2021,13(19). [2]. TANG S, ZHONG R, YIN C, et al. Exposure to High Aerial Ammonia Causes Hindgut Dysbiotic Microbiota and Alterations of Microbiota-Derived Metabolites in Growing Pigs[J]. Front Nutr, 2021,8:689818.

[0082] The results of measuring the intestinal flora of the rats are shown in Tables 3 and 4.

[0083] [Table 3]

[0084] [Table 4]

[0085] NOTE: Different letters abcde indicate differences (p<0.05) between different groups, while groups marked with the same letter indicate no significant difference between these groups.

[0086] When the rat intestinal flora measurement results were analyzed at the phylum level, the relative abundance of the model groups of Firmicutes, Actinobacteria, and Proteobacteria increased significantly, while the comparative group significantly decreased compared to the model group, but still showed a difference compared to the examples. The results of the examples for Actinobacteria and Firmicutes were similar to the results of the blank group, and the abundance of the model group and comparative group of Bacteroidetes decreased significantly, but the example group showed an increasing trend.

[0087] At the species level, three beneficial bacterial strains, namely Bifidobacteria, Lactobacillus and Bacteroides, were significantly improved in the Example group, which was significantly different from the Comparative Example and Model groups. Akkermansia and Roseburia, which are related to weight control, were also significantly improved in the Example group, which indicates to some extent that the Example promotes the growth of beneficial intestinal bacteria and bacterial strains related to fat reduction.

[0088] Example 6 Detection of short-chain fatty acids in rat feces This experimental example detects the content of short-chain fatty acids in rat feces. Figure 7 shows the results of detecting acetic acid in the feces of rats after feeding them room temperature zero sucrose yogurt of the example and the comparative example. Figure 8 shows the results of detecting propionic acid in the feces of rats after feeding them room temperature zero sucrose yogurt of the example and the comparative example. Figure 9 shows the results of detecting butyric acid in the feces of rats after feeding them room temperature zero sucrose yogurt of the example and the comparative example. For detection methods, please refer to the following literature: [1]. MACHIELS K, JOOSSENS M, SABINO J, et al. A decrease of the butyrate-producing species Roseburia hominis and Faecalibacterium prausnitzii defines dysbiosis in patients with ulcerative colitis[J]. Gut, 2014,63(8): 1275-1283. [2]. XIAO S, LIU C, CHEN M, et al. Scutellariae radix and coptidis rhizoma ameliorate glycolipid metabolism of type 2 diabetic rats by modulating gut microbiota and its metabolites[J]. Appl Microbiol Biotechnol, 2020,104(1): 303-317.

[0089] The results shown in Figures 7 to 9 show that compared with the model group, the content of short-chain fatty acids (acetic acid, propionic acid, butyric acid) in the intestines of rats in the example group was significantly increased, and the results of the example group were significantly different from those of the blank group and the comparison group, indicating that the products of the example group have good effects on the production of short-chain fatty acids in the intestines of rats and on body weight control.

[0090] The final point to be mentioned is that the above embodiments are only for illustrating the technical solutions of the present invention, and are not intended to limit the same. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art can still modify the technical solutions described in the above embodiments or make equivalent substitutions for some technical features therein, and these modifications or substitutions will not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. 1. A method for preparing yogurt, comprising: For every 100 g of the yogurt ingredients, honeysuckle extract, prebiotics, and probiotic Bifidobacterium longum BBMN68 were added in the following amounts: (2-10 g): (1-6 g): (1 x 10 ７～８ CFU, e.g., 1 x 10 ７ CFU or 1 x 10 ８ CFU), the prebiotic is inulin or xylooligosaccharides, and preferably the yogurt is shelf-stable sucrose-free yogurt.

2. The method for extracting the honeysuckle extract according to claim 1, characterized in that the honeysuckle extract is obtained by pre-freezing the honeysuckle, freeze-drying the honeysuckle to obtain honeysuckle powder, dissolving the honeysuckle powder in sterile water at a solid-liquid ratio of 20:1 to 30:1 g / L in an insulated tank at 65 to 75°C, infusing for 10 to 20 minutes, and filtering to obtain the honeysuckle extract.

3. 3. The method of claim 2, wherein after the fermentation of the yogurt is completed, the material is preheated to 50-55°C and maintained for 5-10 minutes before sterilization.

4. The method according to any one of claims 1 to 3, wherein the yogurt is prepared by adding a stabilizer, a sweetener, prebiotics, whey protein, and honeysuckle extract to raw milk and fermenting the mixture with Streptococcus thermophilus, Lactobacillus bulgaricus, and Bifidobacterium longum BBMN68 as fermenting strains, wherein the stabilizer is highly esterified pectin, and the sweetener is erythritol or xylitol.

5. The raw materials for the yogurt include 80 to 100 parts by weight of raw milk, 2 to 10 parts by weight of honeysuckle extract, 0.01 to 10 parts by weight of sweetener, 0.5 to 2 parts by weight of whey protein powder, 0.1 to 0.5 parts by weight of highly esterified pectin, 1 to 6 parts by weight of prebiotics, and 1 x 10 effective live bacteria count. ７～１０ CFU / 100g of Lactobacillus bulgaricus, effective viable cell count 1 x 10 ７～１０ CFU / 100g of Streptococcus thermophilus and effective viable cell count 1 x 10 ７～８ 5. The method of claim 4, wherein the Bifidobacterium longum BBMN68 is contained at a concentration of CFU / 100g.

6. Step (1) of filtering raw milk through a reverse osmosis membrane to obtain a retentate and a permeate; The retentate described in step (1) is preheated to 45-55°C, and the sweetener, whey protein powder, prebiotics, and honeysuckle extract are added, stirred uniformly, homogenized, sterilized, and cooled; (2) introducing Streptococcus thermophilus, Lactobacillus bulgaricus and probiotic Bifidobacterium longum BBMN68, fermenting at 40-45°C for 4-8 hours, fermenting until the pH reaches 4.25-4.35, demulsifying, and cooling to obtain a first material; Step (3) of heating the permeate separated in step (1) to 50-55°C, adding a stabilizer, uniformly stirring, homogenizing, sterilizing, and cooling to obtain a second material; 6. The method of claim 5, further comprising: (4) statically mixing the first material obtained in step (2) with the second material obtained in step (3), preheating to 50-60°C and maintaining for 2-5 minutes, then heating to 65-80°C and sterilizing for 15-60 seconds, cooling to 20-25°C, aseptically homogenizing at 20-30 bar, and aseptically filling to obtain shelf-stable sucrose-free yogurt.

7. 7. The method according to claim 6, wherein in step (2), the homogenization conditions before fermentation are 55-65°C and 150-160 bar, the sterilization conditions are 120-130°C and 5-10 s, the cooling temperature is 40-42°C, and the cooling temperature after demulsification is 20-25°C.

8. 7. The method of claim 6, wherein in step (3), after adding the stabilizer and stirring uniformly, the homogenization conditions are 55-65°C and 150-160 bar, the sterilization conditions are 110-120°C and 15s, and the cooling temperature is 20-25°C.

9. A yogurt prepared by the method according to any one of claims 1 to 8.

10. Application of the yogurt according to claim 9 in the production of health food for fat reduction and body shaping.

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