Rose enzyme liquid and rose enzyme beverage for promoting emotional health, and preparation method therefor

Abstract

The present invention relates to the technical field of functional food products. Disclosed are a rose enzyme liquid and rose enzyme beverage for promoting emotional health, and a preparation method therefor. Sour cherry flesh, double-petaled red roses, roselle, and Perillae folium are subjected to a colloid mill, and water is added for extraction and separation to obtain a concentrate. The concentrate is fermented with Lactiplantibacillus plantarum and Bifidobacterium longum to obtain a rose enzyme liquid, and the rose enzyme liquid is compounded with a saffron extract, a Mentha spicata extract, a bamboo leaf flavonoid, etc. to obtain a rose enzyme beverage. The rose, roselle, sour cherry and Perillae folium, which are both medicinal and edible, are selected as raw materials. The raw materials are fermented by means of using specific probiotic strains to effectively release active ingredients, which are then converted into small molecular substances that are easily absorbed by the human body and relieve emotional anxiety. Moreover, functional ingredients comprising the saffron extract, the Mentha spicata extract and the bamboo leaf flavonoid are compounded to effectively relieve stress, regulate daily negative emotions, and promote physical and mental relaxation, and enhance mental well-being. The sour and sweet taste satisfies consumers' pursuit for flavor, taste and efficacy.

Description

A rose enzyme liquid and rose enzyme beverage for promoting emotional well-being and their preparation method Technical Field

[0001] This invention relates to the field of functional food technology, and in particular to a rose enzyme liquid and a rose enzyme beverage that promote emotional health, as well as their preparation methods. Background Technology

[0002] Contemporary people face immense work and life pressures, leading to a growing prevalence and severity of various emotional problems that are gradually impacting daily life and potentially developing into serious mental illnesses. Common emotional problems include anxiety, poor concentration, fatigue, depression, and sleep disorders. Data from the "White Paper on the Mental Health of Urban Residents in China" shows that 73.6% of urban residents are currently in a state of sub-optimal mental health, 16.1% have varying degrees of mental health problems, and only 10.3% are completely mentally healthy. Furthermore, the incidence of mental health issues among urban residents with chronic diseases is extremely high, with only 5.1% achieving mental well-being. The White Paper also points out that mental health is closely related to physical health.

[0003] This shows that a significant number of people are troubled by emotional problems. Moreover, in recent years, functional foods targeting emotional health have emerged on the market, indicating that people are paying more and more attention to their personal emotional health and have a huge demand for improving emotional problems and relieving anxiety.

[0004] Currently, most emotional health-related products on the market are imported, with relatively few functional foods for emotional health available domestically. While some domestic products address sleep problems stemming from emotional health, they primarily focus on regulating sleep and pre-sleep anxiety, lacking specific validation for alleviating daily emotional and stress-related issues. Given the relative scarcity of emotional health products in China, coupled with the increasing pressures of modern life and work, the development of functional foods for emotional health holds significant market potential. Technical issues

[0005] This invention provides a rose enzyme liquid and a rose enzyme beverage that promote emotional health, as well as their preparation method. It uses rose, hibiscus, sour cherry, and perilla, which are both food and medicine, as raw materials. It is formulated with reference to the theory of monarch, minister, assistant and guide in traditional Chinese medicine. Under the fermentation of specific probiotic strains, the effective components in the raw materials are released and transformed into small molecules that are easily absorbed by the human body. After use, it can improve daily emotional anxiety and regulate stress, while meeting consumers' needs for efficacy and flavor. Technical solutions

[0006] To address the aforementioned technical problems, one objective of this invention is to provide a method for preparing a rose enzyme solution that promotes emotional well-being, comprising the following steps:

[0007] (1) After washing and removing impurities from the sour cherry pulp, double-petaled red rose, hibiscus, and perilla leaves, weigh them according to the proportion, mix them, and process them through a colloid mill to obtain a mixed liquid.

[0008] (2) Add water to the mixture for extraction and separation, and then concentrate the extract under vacuum to obtain a concentrated solution;

[0009] (3) Ferment and culture Lactobacillus plantarum and Bifidobacterium longum to obtain a mixed fermentation broth;

[0010] (4) Inoculate the mixed fermentation broth into the concentrated broth and carry out fermentation culture. After centrifugation and filtration, the rose enzyme broth is obtained.

[0011] By adopting the above technical solution, this application selects rose, hibiscus, sour cherry, and perilla, all of which are considered both food and medicine, as raw materials. Rose is sweet and warm in nature, possessing the effects of regulating qi and relieving depression, harmonizing blood and relieving pain. It can regulate qi and blood circulation, soothe the liver and relieve depression. Hibiscus is sour and cool in nature, and is known as the "plant ruby" due to its unique taste and rich nutritional value. Sour cherry contains rich nutrients and a large amount of polyphenols. It contains abundant tryptophan and serotonin, which help improve sleep quality and cognitive function. Perilla is pungent and warm in nature, containing volatile oils and aromatic compounds, which have a certain sedative and soothing effect, helping to relieve negative emotions such as anxiety and tension, promoting relaxation and enhancing mood. This application ferments the raw materials with specific probiotic strains, effectively releasing active ingredients and converting them into small molecules that are easily absorbed by the human body and relieve emotional anxiety. It effectively relieves stress, regulates daily negative emotions, promotes relaxation and mental well-being, and the rose enzyme liquid has a sweet and sour taste, satisfying consumers' pursuit of flavor, taste, and efficacy.

[0012] As a preferred embodiment, in step (1), the mass ratio of rose petals, hibiscus, sour cherry and perilla is (1-10):(1-5):(1-5):(0.5-4).

[0013] As a preferred option, in step (1), the mass ratio of rose petals, hibiscus, sour cherry and perilla is 6:3:3:2.

[0014] As a preferred embodiment, in step (2), water of 5-10 times the volume of the mixture is added to the mixture for several extractions and separations. The extraction temperature is 85-100 ℃ and the extraction time is 0.5-4 h. The extract is mixed evenly and then concentrated under vacuum to obtain a concentrated solution.

[0015] As a preferred embodiment, in step (3), *Lactobacillus plantarum* and *Bifidobacterium longum* are mixed at a mass ratio of (1-3):(1-2), and then inoculated into the culture medium at an inoculum size of 0.5-5%. The mixture is then cultured at 35-38 ℃ for 18-24 h until the pH of the bacterial solution is <4.25, OD >4.00, and the viable count is ≥1.0×10⁻⁶. 9 Fermentation was stopped when the concentration of CFU / mL was reached, yielding a mixed fermentation broth.

[0016] As a preferred option, the mass ratio of *Lactobacillus plantarum* to *Bifidobacterium longum* is 2:1.

[0017] As a preferred option, the viable count of *Lactobacillus plantarum* and *Bifidobacterium longum* is ≥15 billion / g.

[0018] As a preferred embodiment, the culture medium for fermentation in step 3) comprises the following components at the following concentrations: glucose 20-60 g / L, yeast peptone 10-30 g / L, yeast extract 10-35 g / L, L-malic acid 1-5 g / L, citric acid 1-10 g / L, potassium dihydrogen phosphate 1-5 g / L, sodium chloride 0.1-1 g / L, magnesium sulfate 0-0.5 g / L, manganese sulfate 0-0.5 g / L, with the remainder being purified water.

[0019] As a preferred embodiment, the culture medium for fermentation in step 3) comprises the following components at the following concentrations: glucose 30 g / L, yeast peptone 15 g / L, yeast extract 20 g / L, L-malic acid 2 g / L, citric acid 3 g / L, potassium dihydrogen phosphate 2 g / L, sodium chloride 0.6 g / L, magnesium sulfate 0.05 g / L, manganese sulfate 0.01 g / L, with the remainder being purified water.

[0020] As a preferred embodiment, in step (4), the mixed fermentation broth is inoculated into the concentrate at a concentration of 4wt%-10wt% of the concentrate mass. After fermentation for more than 48 hours, the pH value of the fermentation broth is monitored every hour. When the difference between two consecutive pH values ​​is less than 0.1 and the pH is <3.4, ​​fermentation is stopped, and the mixture is centrifuged and filtered to obtain rose enzyme solution.

[0021] As a preferred option, in step (4), the centrifugation speed is 8000-12000 r / min.

[0022] To address the aforementioned technical problems, a second objective of this invention is to provide a rose enzyme liquid that promotes emotional well-being.

[0023] To address the aforementioned technical problems, a third objective of this invention is to provide an application of rose enzyme liquid, which promotes emotional health, in the preparation of food products.

[0024] To address the aforementioned technical problems, the fourth objective of this invention is to provide a rose enzyme beverage that promotes emotional well-being, comprising the following components by mass fraction: 1wt%-10wt% rose enzyme liquid, 0.1wt%-0.5wt% saffron extract, 0.2wt%-1wt% spearmint extract, 0.5wt%-2wt% bamboo leaf flavonoids, 2wt%-10wt% apple concentrate, 0.05wt%-3wt% pectin, 1wt%-15wt% sweetener, 0.01wt%-4wt% acidity regulator, and the balance being water.

[0025] By adopting the above-mentioned approach, saffron extract has the effects of relieving inflammation and anti-oxidation. Spearmint extract is a polyphenol complex containing rosmarinic acid, salvianolic acid, shikonin, and caffeic acid. The natural polyphenol antioxidants composed of these components can reduce oxidative stress in the brain, increase neurotransmitter levels, protect neurons, and promote the growth of new neurons. Bamboo leaf flavonoids are physiologically active bioflavonoids extracted from bamboo, which can improve brain tissue nutrition and microcirculation, and have anti-depressant and memory-improving functions. When these three are combined with rose enzyme liquid fermented by probiotics, they can significantly improve emotional stress, soothe mood, save on the amount of functional ingredients required, and reduce production costs.

[0026] As a preferred embodiment, the sweetener comprises erythritol at 1 wt%-15 wt% of the rose enzyme beverage and sucralose at 0.001 wt%-0.1 wt% of the beverage, and the acidity adjuster comprises citric acid at 0.01 wt%-0.5 wt% of the rose enzyme beverage and sodium citrate at 0.01 wt%-4 wt% of the beverage.

[0027] To address the aforementioned technical problems, the fifth objective of this invention is to provide a method for preparing a rose enzyme beverage that promotes emotional well-being, comprising the following steps:

[0028] 1) Mix erythritol and pectin evenly, add half water and stir and heat until completely dissolved to obtain solvent A;

[0029] 2) Dissolve saffron extract, spearmint extract, and bamboo leaf flavonoids in the remaining water to obtain solution B;

[0030] 3) Add solution B to solution A, then add rose enzyme solution, apple concentrate, sucralose, citric acid and sodium citrate. Stir well to obtain a rose enzyme beverage that promotes mood health. Beneficial effects

[0031] Compared with the prior art, the present invention has the following beneficial effects:

[0032] 1. This application uses rose, hibiscus, sour cherry and perilla, which are both food and medicine, as raw materials. The raw materials are fermented by specific probiotic strains to effectively release active ingredients and transform them into small molecules that are easily absorbed by the human body and relieve emotional anxiety. It effectively relieves stress, regulates daily negative emotions, promotes physical and mental relaxation and mental pleasure. Moreover, the rose enzyme liquid has a sweet and sour taste, which satisfies consumers' pursuit of flavor and efficacy.

[0033] 2. Saffron extract, spearmint extract, and bamboo leaf flavonoids all have antioxidant effects, promote neuronal growth, and enrich brain tissue nutrition. When these three are combined with rose enzyme liquid fermented by probiotics, they can significantly improve emotional stress, soothe mood, save on the amount of functional ingredients needed, and reduce production costs. Embodiments of the present invention

[0034] Type the description paragraph of embodiments of the present invention here.

[0035] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. 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 skilled in the art without creative effort are within the scope of protection of the present invention.

[0036] In the embodiments, unless otherwise specified, the experimental methods used are conventional methods, and the materials and reagents used are commercially available unless otherwise specified. The specific sources of the raw materials used in this invention are as follows: the probiotics (Lactobacillus plantarum and Bifidobacterium longum) used are commercially available bacterial powders. Lactobacillus plantarum JYLP-326, Bifidobacterium longum JBLC-141, Lactobacillus rhamnosus JYLR-127 and Bifidobacterium animalis JYBR-190 were all purchased from Shandong Zhongke Jiayi Bioengineering Co., Ltd., with a live bacteria count of 15 billion / g. Saffron extract was purchased from Guangzhou Kanglai Food Additives Co., Ltd., spearmint extract was purchased from Kemin (China) Technology Co., Ltd., and bamboo leaf flavonoids were purchased from Hangzhou Muhua Biotechnology Co., Ltd.

[0037] Example 1

[0038] A rose enzyme liquid that promotes emotional well-being is prepared by the following steps:

[0039] 1) Raw material pretreatment: Wash, remove impurities, pit and destem the sour cherries, wash and remove impurities the fresh double-petal red roses, hibiscus, and perilla leaves, weigh and mix them together with the sour cherry pulp in the specified proportions, and process them through a colloid mill to obtain a mixed liquid; the mass ratio of rose petals, hibiscus, sour cherries and perilla leaves is 2:2:5:3.5;

[0040] 2) Preparation of concentrated extract: Add water at 10 times the volume of the ground mixture to the mixture for the first extraction and separation at 95℃ for 3 hours to obtain extract and residue; add water at 8 times the volume of the residue to the residue for the second extraction and separation at 95℃ for 1 hour. Mix the two extracts evenly and concentrate them three times under vacuum to obtain concentrated extract.

[0041] 3) Strain activation: *Lactobacillus plantarum* JYLP-326 and *Bifidobacterium longum* JBLC-141 were mixed at a mass ratio of 2:1, dissolved in sterile water, and then inoculated into an optimized culture medium at a strain inoculation amount of 3 wt%. The culture was carried out at 37℃ for 18-24 hours until the pH of the culture was <4.25, OD >4.00, and the viable cell count was ≥1.0 × 10⁻⁶. 9 Fermentation was stopped when the concentration of CFU / mL was reached. Fermentation was then complete, and a mixed fermentation broth was obtained.

[0042] The optimized culture medium consisted of 30 g / L glucose, 15 g / L yeast peptone, 20 g / L yeast extract, 2 g / L L-malic acid, 3 g / L citric acid, 2 g / L potassium dihydrogen phosphate, 0.6 g / L sodium chloride, 0.05 g / L magnesium sulfate, 0.01 g / L manganese sulfate, with the remainder being purified water.

[0043] 4) Fermentation: Inoculate the mixed fermentation broth from step 3) into the concentrated broth from step 2) for fermentation. The inoculation amount of the strain is 7% by mass. Incubate at 37℃ for 60±4h. Starting from 48h of fermentation, monitor the pH value of the fermentation broth every hour. When the difference between two consecutive pH values ​​is less than 0.1 and the pH is <3.4, ​​stop the fermentation. After fermentation, the fermentation broth is centrifuged and filtered at a speed of 10000r / min to obtain the rose enzyme solution.

[0044] Example 2

[0045] A rose enzyme liquid that promotes emotional health is prepared in the same way as Example 1, with the same steps, reagents and process parameters. The difference is that in step 1), the mass ratio of rose petals, hibiscus, sour cherry and perilla leaves is 4:4:2:3.

[0046] Example 3

[0047] A rose enzyme liquid that promotes emotional health is prepared in the same way as Example 1, with the same steps, reagents and process parameters. The difference is that in step 1), the mass ratio of rose petals, hibiscus, sour cherry and perilla leaves is 6:3:3:2.

[0048] Example 4

[0049] A rose enzyme liquid that promotes emotional health is prepared in the same way as Example 1, with the same steps, reagents and process parameters. The difference is that in step 1), the mass ratio of rose petals, hibiscus, sour cherry and perilla leaves is 7:2:4:1.5.

[0050] Example 5

[0051] A rose enzyme liquid that promotes emotional health is prepared in the same way as Example 1, with the same steps, reagents and process parameters. The difference is that in step 1), the mass ratio of rose petals, hibiscus, sour cherry and perilla leaves is 9:4.5:1:1.

[0052] Comparative Example 1

[0053] A rose enzyme liquid that promotes emotional health is prepared using the same method and reagents and process parameters as in Example 3, except that double-petaled red roses are not added.

[0054] Comparative Example 2

[0055] A rose enzyme liquid that promotes emotional health is prepared using the same method and reagents and process parameters as in Example 3, except that roselle is not added.

[0056] Comparative Example 3

[0057] A rose enzyme liquid that promotes emotional health is prepared using the same method and reagents and process parameters as in Example 3, except that sour cherries are not added.

[0058] Comparative Example 4

[0059] A rose enzyme liquid that promotes emotional health is prepared using the same method and reagents and process parameters as in Example 3, except that perilla leaves are not added.

[0060] Comparative Example 5

[0061] A rose concentrate that promotes emotional well-being is prepared by the following steps:

[0062] 1) Raw material pretreatment: Wash, remove impurities, pit and stem the sour cherries, wash and remove impurities the fresh double-petal red roses, hibiscus, and perilla leaves, and process them together with the sour cherry pulp through a colloid mill. The mass ratio of rose petals, hibiscus, sour cherries and perilla leaves is 6:3:3:2.

[0063] 2) Preparation of concentrated extract: Add water at 10 times the volume of the ground mixture to the mixture for the first extraction and separation. The extraction temperature is 95℃ and the extraction time is 3h to obtain the initial extract and residue. Add water at 8 times the volume of the residue to the residue for the second extraction and separation. The extraction temperature is 95℃ and the extraction time is 1h to obtain the re-extract. Mix the two extracts evenly and concentrate them three times under vacuum to obtain the concentrated extract.

[0064] Comparative Example 6

[0065] A rose enzyme liquid that promotes emotional health is prepared in the same way as Example 3, with the same steps, reagents and process parameters. The difference is that in step 3), Lactobacillus rhamnosus JYLR-127 is used to replace Lactobacillus plantarum JYLP-326 in an equal amount, and Bifidobacterium animalis JYBR-190 is used to replace Bifidobacterium longum JBLC-141 in an equal amount.

[0066] Examples 6-10

[0067] A rose enzyme beverage that promotes mood health, the components and their contents are shown in Table 1 below, and its preparation method includes the following steps:

[0068] 1) After mixing erythritol and pectin evenly, add half of the water, stir and heat to 85°C until completely dissolved;

[0069] 2) Dissolve the saffron extract, spearmint extract, and bamboo leaf flavonoids in the remaining water;

[0070] 3) Add the solution obtained in step 2) to the solution obtained in step 1) of the pectin solution, then add rose enzyme liquid, apple concentrate, sucralose, citric acid and sodium citrate. Stir well to obtain a rose enzyme beverage that promotes mood health.

[0071] Table 1 - Content of each component in the rose enzyme beverage of Examples 6-10

[0072]

[0073] Comparative Example 7

[0074] A rose enzyme beverage that promotes emotional well-being is made with the same steps, reagents, and process parameters as in Example 8, except that an equal amount of water is used instead of rose enzyme liquid.

[0075] Comparative Example 8

[0076] A rose enzyme beverage that promotes emotional well-being is made using the same steps, reagents, and process parameters as in Example 8, except that the saffron extract is replaced with an equal amount of water.

[0077] Comparative Example 9

[0078] A rose enzyme beverage that promotes emotional well-being is made using the same steps, reagents, and process parameters as in Example 8, except that spearmint extract is replaced with an equal amount of water.

[0079] Comparative Example 10

[0080] A rose enzyme beverage that promotes emotional well-being is made using the same steps, reagents, and process parameters as in Example 8, except that bamboo leaf flavonoids are replaced with an equal amount of water.

[0081] Comparative Example 11

[0082] A rose enzyme beverage that promotes emotional well-being is made with the same steps, reagents, and process parameters as in Example 8. The difference is that the content of rose enzyme liquid is 0.05 kg, the content of saffron extract is 2 kg, the content of spearmint extract is 0.05 kg, and the content of bamboo leaf flavonoids is 4.645 kg.

[0083] Performance testing

[0084] 1. Mouse tail-suspended depression model experiment:

[0085] This experiment verifies the antidepressant effect of the rose enzyme solution described in Examples 1-10 and Comparative Examples 1-11. The specific experimental procedure is as follows: Mice were randomly divided into 23 groups, with 10 mice in each group (half male and half female): Examples 1-10, Comparative Examples 1-11, a normal group, and a model group. Mice in the model group, Examples 1-10, and Comparative Examples 1-11 were randomly given two different stimuli daily, including: 24-hour water deprivation, 24-hour fasting, inclined cage, electric shock, 4-hour restraint, wet cage, and reversed day-night cycle. This continued for 8 weeks. After the depressive phenotype appeared, the depressed mice were administered the drug via gavage. The samples from Examples 1-5 and Comparative Examples 1-6 were diluted with water to a 10 wt% concentration. The dosage for Examples 1-10 and Comparative Examples 1-11 was 0.4 mL per mouse, while the normal group and model group received an equal volume of physiological saline. Each mouse was administered the drug twice daily via gavage for 10 consecutive days.

[0086] Tail suspension test: Adhesive tape was attached to the tail of the mouse about 1 cm away from the tail, and the mouse was suspended upside down in the box with its head about 10 cm from the bottom of the box. The mouse was suspended for 6 minutes. At the beginning of the suspension, the mouse would struggle and shake, and then it would give up struggling. The duration of the mouse remaining still within 2-6 minutes was recorded. The test results are shown in Table 2.

[0087] 2. Measurement of serum stress factor levels:

[0088] After the tail suspension test, the mice were euthanized, and blood was routinely collected from the orbital cavity. After standing at room temperature for 2 hours, the blood was centrifuged at 3500 rpm / min and 4℃. The serum was collected, and the levels of adrenocorticotropic hormone (ACTH) and cortisol were measured. The results are shown in Table 2.

[0089] Table 2 - Tail suspension rest time and ACTH and cortisol content of the products prepared in Examples 1-10 and Comparative Examples 1-11 of this application after mouse tail suspension depression model test.

[0090]

[0091] Table 2 shows that compared with the model group and comparative examples 1-6, the tail immobility time of mice in each of the Examples 1-5 groups was reduced, indicating that the rose enzyme solution obtained in each of the Examples 1-5 groups can effectively alleviate anxiety and promote emotional health in mice. Furthermore, the tail immobility time of mice in Example 3 was the shortest, and close to that of the normal group, indicating that the rose enzyme solution in Example 3 represents the optimal combination of raw materials and fermentation strains, resulting in the best anxiety-relieving effect. In Comparative Example 5, the tail immobility time of mice was reduced to some extent compared to the model group, but significantly longer than that in Examples 1-5. This suggests that while double-petaled red roses, hibiscus, sour cherry, and perilla leaves have some effect on improving emotional health, they do not produce a significant improvement effect. The rose enzyme solution produced by the fermentation process in this application has a better effect on improving emotional health. Compared to the model group, the tail immobility time of mice in Comparative Example 6 was reduced to some extent, but compared to Examples 1-5, the tail immobility time of mice was significantly longer. This indicates that the combination of double-petaled red rose, hibiscus, sour cherry, and perilla leaf, after fermentation by other strains, also has a certain effect, but it cannot produce a significant improvement effect. The product of co-fermentation of double-petaled red rose, hibiscus, sour cherry, and perilla leaf by Lactobacillus plantarum JYLP-326 and Bifidobacterium longum JBLC-141 in this application has the best effect on improving emotional health.

[0092] Table 2 shows that compared with comparative examples 7-11 and the model group, the tail-suspended immobility time of mice in each of the Examples 6-10 groups was reduced, indicating that the enzyme beverages obtained in each of the Examples 6-10 groups can effectively alleviate anxiety and promote emotional health in mice. Compared with comparative examples 7-11 groups, the tail-suspended immobility time of mice in Examples 6-10 groups was significantly reduced, and the tail-suspended immobility time of mice in Example 8 was the shortest, and close to that of mice in the normal group, indicating that this example is the optimal combination and has the best effect in relieving anxiety. The tail-suspended immobility time of mice in comparative examples 7-11 was significantly reduced compared with the model group, indicating that the rose enzyme liquid, saffron extract, spearmint extract, and bamboo leaf flavonoids in the enzyme beverage all have a certain effect on improving emotional health. However, the absence of raw materials or the addition of too much or too little of individual components will significantly affect the effect of relieving mood and anxiety.

[0093] ACTH and cortisol are stress factors closely related to the pathogenesis of depression. As shown in Table 2, ACTH and cortisol are positively correlated with the tail suspension resting time of mice. Compared with the model group and the control groups 1-6, the levels of ACTH and cortisol in the serum of mice in each of the Examples 1-5 groups were reduced, indicating that the rose enzyme liquid obtained in each of the Examples 1-5 groups can effectively alleviate the anxiety of mice and promote emotional health. Moreover, the levels of ACTH and cortisol in the serum of mice in Example 3 were the lowest and close to those in the serum of normal mice, indicating that the rose enzyme liquid in Example 3 is the best combination of raw materials and fermentation strains, and has the best effect in relieving anxiety.

[0094] Compared with comparative groups 7-11 and the model group, the levels of ACTH and cortisol in the serum of mice in each of the Examples 6-10 groups were reduced, indicating that the enzyme beverages obtained in each of the Examples 6-10 groups can effectively alleviate the anxiety of mice and promote emotional health. In addition, the mice in Example 8 had the shortest tail suspension immobility time and the levels of ACTH and cortisol in the serum of mice in the normal group were close to those in the normal group, indicating that this example is the best combination and has the best effect in relieving anxiety.

[0095] 3. Acute toxicity test (MTD method):

[0096] Experimental Procedure: Mice weighing 18-22 g, half male and half female, were selected and divided into a blank control group and groups 1-10 (n=10 per group). The use of experimental animals followed the regulations for experimental animal use. Animals were housed with a 12h / 12h light / dark cycle, a temperature of 23±1℃, and a humidity of 50±10%, with free access to food and water. All test mice were fasted overnight for 4-6 hours before the experiment, but water was not restricted. For the formal experiment, the gavage dose was 500 ml / (kg·md) (approximately 30 times the daily adult dose), concentrated to a certain proportion before testing. Based on the maximum gavage concentration, gavage volume, and safety factor of the sample, the test substance dose was set at 20.0 mL / kg BW, i.e., 0.4 mL / mouse per gavage. The test sample was administered twice a day at 6-hour intervals (the blank control group received purified water). The test substance was also administered orally for 3 days. After the administration period, mice were observed for 14 consecutive days, and the signs of poisoning and mortality were recorded.

[0097] Experimental results: No deaths or abnormalities occurred in mice under normal feeding conditions within 14 days after administration. The experiment proved that the acute oral LD50 of the enzyme beverages in Examples 1-10 to mice was >500 mL / (kg·md), indicating that this rose enzyme liquid and rose enzyme beverage have no toxic side effects and can be used safely.

[0098] 4. Food tasting test:

[0099] This study used the Positive Mood State Scale (POMS) to evaluate the effectiveness of the enzyme beverage in relieving anxiety and improving mood.

[0100] Experimental Procedure: Forty volunteers aged 20-60 years were selected, with an equal number of men and women. The participants exhibited varying degrees of emotional problems such as insomnia, anxiety, poor concentration, and lethargy. The volunteers were randomly assigned to two groups: a competitor group and an Example 8 group, with 20 participants in each group. The competitor group consisted of the following components by mass fraction: 0.1% γ-aminobutyric acid (GABA), 0.3% tea theanine, 0.2% hops extract, 0.045% vitamin C, 8% sweetener, 0.1% flavoring, and the remainder being water. Volunteers consumed either the competitor group's product (100ml / bottle) or the enzyme beverage prepared in Example 8 (100ml / bottle) once daily for four weeks, during which time their diet and lifestyle remained unchanged. Before, during, and after the food tasting, participants took a questionnaire on emotional issues as shown in Table 3. Questions 1, 8, 15, 21, 28, and 35 in Table 3 were used to assess tension; questions 2, 9, 16, 22, 29, 36, and 37 were used to assess anger; questions 3, 10, 17, 23, and 30 were used to assess fatigue; questions 4, 11, 18, 24, 31, and 38 were used to assess depression; questions 5, 12, 19, 25, 32, and 39 were used to assess energy; questions 6, 13, 20, 26, and 33 were used to assess panic; and questions 7, 14, 27, 34, and 40 were used to assess self-esteem. The assessment results are shown in Table 4.

[0101] POMS scoring method: "almost none" is 0 points, "somewhat" is 1 point, "moderate" is 2 points, "quite a lot" is 3 points, and "very little" is 4 points.

[0102] The Positive Mood State Scale (POMS) comprises 7 assessment dimensions. The Total Mood Disturbance (TMD) score is calculated as follows: (Total score for 5 negative emotions: tension, anger, depression, panic) - (Total score for 2 positive emotions: energy, self-esteem) + 100. A higher TMD score indicates a worse mood state, and vice versa.

[0103] Table 3 - Evaluation of the POMS (Positive Mood State Scale) Questionnaire Survey

[0104]

[0105]

[0106] Table 4 - Evaluation results of Example 8 of this application and the competing products on the Positive Mood State Scale (POMS).

[0107]

[0108] As can be seen from the questionnaire evaluation results in Table 4, after a 4-week trial, the total TMD scores of both the competitor group and the Example 8 group decreased, but the decrease in the competitor group's score was smaller, indicating that the improvement effect was not significant; while the score of the Example 8 group decreased significantly, showing a highly significant difference compared with before the trial (P < 0.01). This indicates that Example 8 of this application has a highly significant effect on improving anxiety and relieving stress.

[0109] The specific embodiments described above further illustrate the purpose, technical solution, and beneficial effects of the present invention. It should be understood that the above descriptions are merely specific embodiments of the present invention and are not intended to limit the scope of protection of the present invention. In particular, it should be noted that any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention for those skilled in the art.

Claims

1. A method for preparing a rose enzyme liquid that promotes emotional well-being, characterized in that, Includes the following steps: (1) After washing and removing impurities from the sour cherry pulp, double-petaled red rose, hibiscus, and perilla leaves, the mixture was processed through a colloid mill to obtain a mixed liquid. (2) Add water to the mixture for extraction and separation, and then concentrate the extract under vacuum to obtain a concentrated solution; (3) Ferment and culture Lactobacillus plantarum and Bifidobacterium longum to obtain a mixed fermentation broth; (4) Inoculate the mixed fermentation broth into the concentrated broth and carry out fermentation culture. After centrifugation and filtration, the rose enzyme broth is obtained.

2. The method for preparing a rose enzyme liquid that promotes emotional health as described in claim 1, characterized in that, In step (1), the mass ratio of rose petals, hibiscus, sour cherry and perilla is (1-10): (1-5): (1-5): (0.5-4).

3. The method for preparing a rose enzyme liquid that promotes emotional health as described in claim 1, characterized in that, In step (2), water of 5-10 times the volume of the mixture is added to the mixture for several extractions and separations. The extraction temperature is 85-100 ℃ and the extraction time is 0.5-4 h. The extract is mixed evenly and then concentrated under vacuum to obtain a concentrated solution.

4. The method for preparing a rose enzyme liquid that promotes emotional health as described in claim 1, characterized in that, In step (3), *Lactobacillus plantarum* and *Bifidobacterium longum* are mixed at a mass ratio of (1-3):(1-2), and then inoculated into the culture medium at an inoculum volume of 0.5-5%. The mixture is then cultured at 35-38 ℃ for 18-24 h until the pH of the bacterial solution is <4.25, OD >4.00, and the viable count is ≥1.0×10⁻⁶. 9 Fermentation was stopped when the concentration of CFU / mL was reached, yielding a mixed fermentation broth.

5. The method for preparing a rose enzyme liquid that promotes emotional health as described in claim 1, characterized in that, In step (4), the mixed fermentation broth is inoculated into the concentrate at a concentration of 4wt%-10wt% of the concentrate mass. After fermentation for more than 48 hours, the pH value of the fermentation broth is monitored every hour. When the difference between two consecutive pH values ​​is less than 0.1 and the pH is less than 3.4, fermentation is stopped, centrifuged and filtered to obtain rose enzyme liquid.

6. A rose enzyme liquid for promoting emotional health prepared using the method for preparing rose enzyme liquid for promoting emotional health as described in any one of claims 1-5.

7. The application of the rose enzyme liquid for promoting emotional health as described in claim 6 in the preparation of food products.

8. A rose enzyme beverage that promotes emotional well-being, characterized in that, The rose enzyme liquid for promoting emotional health as described in claim 6 comprises the following components by mass fraction: 1 wt%-10 wt% rose enzyme liquid, 0.1 wt%-0.5 wt% saffron extract, 0.2 wt%-1 wt% spearmint extract, 0.5 wt%-2 wt% bamboo leaf flavonoids, 2 wt%-10 wt% apple concentrate, 0.05 wt%-3 wt% pectin, 1 wt%-15 wt% sweetener, 0.01 wt%-4 wt% acidity regulator, and the balance being water.

9. A rose enzyme beverage for promoting emotional well-being as described in claim 8, characterized in that, The sweetener comprises erythritol at 1 wt%-15 wt% of the rose enzyme beverage and sucralose at 0.001 wt%-0.1 wt% of the beverage, and the acidity adjuster comprises citric acid at 0.01 wt%-0.5 wt% of the rose enzyme beverage and sodium citrate at 0.01 wt%-4 wt% of the beverage.

10. A method for preparing a rose enzyme beverage for promoting emotional well-being as described in claim 9, characterized in that, Includes the following steps: 1) Mix erythritol and pectin evenly, add half water and stir and heat until completely dissolved to obtain solvent A; 2) Dissolve saffron extract, spearmint extract, and bamboo leaf flavonoids in the remaining water to obtain solution B; 3) Add solution B to solution A, then add rose enzyme solution, apple concentrate, sucralose, citric acid and sodium citrate. Stir well to obtain a rose enzyme beverage that promotes mood health.

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