A composition containing a plantain fruit extract, a drink, and a preparation method and application thereof

Abstract

This invention discloses a composition containing banana fruit extract, a beverage thereof, its preparation method, and its application, belonging to the field of biotechnology. This invention combines banana fruit extract, apple and blueberry extract, cherry extract, and sweet potato leaf extract, which have sleep-improving effects; then, it is combined with water and other excipients to prepare a beverage. This beverage is made entirely from natural extracts, can be taken long-term, has no toxic side effects, and has a significant sleep-improving effect.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, and particularly relates to a composition containing banana fruit extract, a beverage thereof, its preparation method and application. Background Technology

[0002] Currently, sleep disorders have become a health problem troubling a large number of people, manifesting as various symptoms such as difficulty falling asleep, light sleep, frequent awakenings, and early awakenings. Long-term sleep disorders can not only lead to daytime lethargy and poor concentration, but may also induce a series of chain health problems such as neurasthenia, anxiety, high blood pressure, and weakened immunity.

[0003] Currently, sleep improvement products are mainly divided into two categories: chemically synthesized drugs and natural product-based functional products. Chemically synthesized drugs, such as benzodiazepines and non-benzodiazepines, while effective quickly, are prone to dependence and tolerance with long-term use, and may be accompanied by side effects such as dizziness, drowsiness, and memory loss, resulting in poor safety. Furthermore, existing natural sleep improvement products generally suffer from low absorption efficiency of active ingredients, slow effects, and lack of specificity. Therefore, developing sleep improvement products that are naturally derived, highly safe, have no side effects, and offer stable efficacy has become a hot research topic in the industry. Summary of the Invention

[0004] In view of this, the purpose of the present invention is to provide a composition containing banana fruit extract, a beverage thereof, a method for preparing the same, and an application thereof. The composition and beverage have significant effects in improving sleep, can be consumed long-term, have stable effects, and have no side effects.

[0005] To achieve the above-mentioned objectives, the present invention provides the following technical solution: The present invention provides a composition containing banana fruit extract, comprising the following raw materials in parts by weight: 45-65 parts banana fruit extract, 25-45 parts apple blueberry extract, 15-35 parts cherry extract and 25-45 parts sweet potato leaf extract.

[0006] This invention provides a method for preparing the composition, comprising the following steps: mixing plantain fruit with ethanol, sonicating, centrifuging to collect the centrifuged liquid, concentrating to obtain plantain fruit extract; crushing apples and blueberries into pulp, adding pectinase and cellulase for enzymatic hydrolysis, inactivating the enzymes, then adding yeast and Lactobacillus plantarum YLP-326 for fermentation, centrifuging to collect the centrifuged liquid, concentrating to obtain apple-blueberry extract; crushing cherries, adding ethanol for supercritical CO2 extraction, freeze-drying the extract to obtain cherry extract; sonicating sweet potato leaf powder with water, adding Lactobacillus fermentum and Streptococcus thermophilus for fermentation, centrifuging to collect the centrifuged liquid, concentrating to obtain sweet potato leaf extract.

[0007] Preferably, the temperature of the ultrasonic mixing of banana fruit and ethanol is 50-60℃, the power is 150-250W, the time is 30-50min, and the number of times is 1-3.

[0008] Preferably, the enzyme activity ratio of pectinase to cellulase is 1:0.5-1.5; the total enzyme activity of pectinase and cellulase is 200-300 U / g of the total weight of apples and blueberries; and the enzymatic hydrolysis temperature is 40-50℃ and the time is 3-10h.

[0009] Preferably, the live count ratio of the yeast to Lactobacillus plantarum YLP-326 is 1-3:1; the total live count of the yeast and Lactobacillus plantarum YLP-326 is 0.5-1.5 × 10⁻⁶. 8 The fermentation solution was CFU / mL; the fermentation temperature was 27-35℃ and the time was 24-48h.

[0010] Preferably, the mass-to-volume ratio of the cherry to ethanol is 1g:4-7mL; the extraction pressure is 19-20MPa, the temperature is 38-42℃, the time is 100-140min, and the CO2 flow rate is 12-18kg / h.

[0011] Preferably, the ratio of viable Lactobacillus fermentum to Streptococcus thermophilus is 1:1-3; the total viable count of Lactobacillus fermentum and Streptococcus thermophilus is 3.5-6.5 × 10⁻⁶. 7 The concentration of cfu / mL in the ultrasonic extract was used; the fermentation temperature was 27-35℃ and the time was 24-48h.

[0012] This invention provides a beverage containing banana fruit extract, comprising the composition described above or the composition obtained by the preparation method described above.

[0013] Preferably, it also includes water, glucose syrup, vitamin C, and sodium citrate.

[0014] The present invention provides the use of the composition or the composition obtained by the preparation method or the beverage in the preparation of sleep-improving products.

[0015] The beneficial effects of this invention are: This invention is the first to combine banana fruit extract, apple and blueberry extract, cherry extract, and sweet potato leaf extract. The active ingredients in each extract interact with each other, complementing and enhancing each other's effects. Compared with single ingredients and other combinations, it has a significant effect on improving sleep. When this composition is prepared into a beverage, it is more suitable for the general public. Moreover, the active ingredients are all naturally extracted, with stable effects and reliable safety. Detailed Implementation

[0016] This invention provides a composition containing plantain fruit extract, comprising the following ingredients in parts by weight: 45-65 parts plantain fruit extract, 25-45 parts apple-blueberry extract, 15-35 parts cherry extract, and 25-45 parts sweet potato leaf extract; preferably comprising the following ingredients in parts by weight: 50-60 parts plantain fruit extract, 30-40 parts apple-blueberry extract, 20-30 parts cherry extract, and 30-40 parts sweet potato leaf extract. This invention targets extracts obtained from each ingredient using specific methods that contain active ingredients that help improve sleep, and the synergistic effect of these ingredients results in a more significant sleep-improving effect than extracts from a single ingredient.

[0017] This invention provides a method for preparing the composition, comprising the following steps: mixing plantain fruit with ethanol, sonicating, centrifuging to collect the centrifuged liquid, concentrating to obtain plantain fruit extract; crushing apples and blueberries into pulp, adding pectinase and cellulase for enzymatic hydrolysis, inactivating the enzymes, then adding yeast and Lactobacillus plantarum YLP-326 for fermentation, centrifuging to collect the centrifuged liquid, concentrating to obtain apple-blueberry extract; crushing cherries, adding ethanol for supercritical CO2 extraction, freeze-drying the extract to obtain cherry extract; sonicating sweet potato leaf powder with water, adding Lactobacillus fermentum and Streptococcus thermophilus for fermentation, centrifuging to collect the centrifuged liquid, concentrating to obtain sweet potato leaf extract.

[0018] In this invention, the ultrasonic treatment of banana fruit and ethanol is performed at a temperature of 50-60℃, a power of 150-250W, a time of 30-50min, and 1-3 times; preferably, the ultrasonic temperature is 52-58℃, the power is 160-230W, the time is 32-48min, and the number of times is 2. The concentration of ethanol in this invention is 70-80 v / v%, preferably 72-78%. The mass-to-volume ratio of banana fruit to ethanol in this invention is 1g:20-30mL, preferably 1g:22-28mL. This invention uses an ultrasonic extraction process of banana fruit with ethanol. The cavitation effect of ultrasound can efficiently destroy the cell wall of banana fruit and promote the dissolution of active ingredients. Ethanol, as a solvent, has better solubility for polar components such as phenols and tryptophan than water, avoiding interference from polysaccharide impurities that are easily introduced during water extraction. Subsequent centrifugation to remove fiber residue and concentration further increases the concentration of sleep-improving components in the extract, thereby enhancing the effect of regulating sleep rhythm.

[0019] In this invention, the mass ratio of apples to blueberries is 1:0.5-2.5, preferably 1:0.8-1.5. The enzyme activity ratio of pectinase to cellulase is 1:0.5-1.5, preferably 1:0.8-1.3; the total enzyme activity of pectinase and cellulase is 200-300 U / g of the total weight of apples and blueberries, preferably 220-280 U / g of the total weight of apples and blueberries; the enzymatic hydrolysis temperature is 40-50℃ and the time is 3-10 h, preferably 42-48℃ and 4-8 h. The enzymatic hydrolysis process of this invention degrades pectin and cellulose in the fruit pulp, breaks down cell wall barriers, and allows the sleep-promoting components in apples and blueberries to be fully released. Compared with direct pulverization or water extraction, it is easier to obtain active ingredients that help improve sleep. The pectinase and cellulase mentioned in this invention are commercially available products well known in the art.

[0020] In this invention, the live cell ratio of the yeast and Lactobacillus plantarum YLP-326 is 1-3:1, preferably 1.5-2.5:1; the total live cell count of the yeast and Lactobacillus plantarum YLP-326 is 0.5-1.5 × 10⁻⁶. 8 The cfu / mL enzymatic hydrolysate is preferably prepared with a total viable count of 0.8-1.2 × 10⁻⁶. 8 CFU / mL enzymatic hydrolysate. The fermentation temperature of this invention is 27-35℃, and the time is 24-48h, preferably 28-33℃, and the time is 26-46h. This invention uses yeast and Lactobacillus plantarum YLP-326 to simultaneously ferment apples and blueberries, which can obtain active ingredients that calm the central nervous system and shorten the sleep latency. At the same time, the fermentation process breaks down large molecules into smaller molecules, reduces the digestive burden on the gastrointestinal tract, improves the bioavailability of active ingredients, and allows the ingredients to act on the target site more quickly, thereby improving sleep. The yeast used in this invention is from Ningbo Mingzhou Biotechnology Co., Ltd., with the product number BMZ126199; the Lactobacillus plantarum YLP-326 is from Minsheng Zhongke Jiayi (Shandong) Bioengineering Co., Ltd.

[0021] In this invention, the volume-to-mass ratio of cherry to ethanol is 1g:4-7mL, preferably 1g:5-6.5mL; the extraction pressure is 19-20MPa, the temperature is 38-42℃, the time is 100-140min, and the CO2 flow rate is 12-18kg / h; preferably, the pressure is 19.2-19.8MPa, the temperature is 39-41℃, the time is 110-130min, and the CO2 flow rate is 14-16kg / h. This invention uses a supercritical CO2 extraction process with ethanol, which can efficiently extract sleep-aiding components from cherries while avoiding the degradation of active ingredients caused by high-temperature extraction. The resulting cherry extract can more precisely regulate the sleep-wake cycle.

[0022] In this invention, the weight ratio of sweet potato leaf powder to water is 1:15-25, preferably 1:18-23. The ultrasonic mixing of the sweet potato leaf powder and water is performed at a temperature of 40-50℃, a power of 250-350W, a time of 45-55 minutes, and 1-3 times, preferably at a temperature of 42-48℃, a power of 260-340W, a time of 48-53 minutes, and 2 times. This invention utilizes ultrasonic cavitation to disrupt the cell structure of sweet potato leaves, releasing flavonoids and polyphenols from the cells into the aqueous phase system, providing sufficient substrate for microbial fermentation and facilitating the acquisition of sleep-enhancing active ingredients.

[0023] In this invention, the ultrasonic extract of sweet potato leaves is fermented. The ratio of viable Lactobacillus fermentation to Streptococcus thermophilus added in this invention is 1:1-3, preferably 1:1.5-2.5; the total viable count of the Lactobacillus fermentation and Streptococcus thermophilus is 3.5-6.5 × 10⁻⁶. 7 CFU / mL ultrasonic extraction buffer, preferably with a total viable bacterial count of 4-6 × 10⁻⁶. 7 The fermentation solution was extracted using ultrasonic extraction at a concentration of CFU / mL. The fermentation temperature was 27-35℃, and the time was 24-48 hours, preferably 29-32℃, and the time was 26-46 hours. This invention utilizes two strains, *Lactobacillus fermentatus* and *Streptococcus thermophilus*, to co-ferment sweet potato leaves. This not only converts macromolecular active ingredients into more easily absorbed small molecule derivatives but also synthesizes additional active ingredients that indirectly improve sleep quality by regulating nerve function through the gut-brain axis. The *Lactobacillus fermentatus* strain used in this invention is from Ningbo Mingzhou Biotechnology Co., Ltd., with product number BMZ134605; the *Streptococcus thermophilus* strain is from Ningbo Mingzhou Biotechnology Co., Ltd., with product number BMZ124652.

[0024] This invention provides a beverage containing banana fruit extract, comprising the aforementioned composition or the composition obtained by the aforementioned preparation method. The beverage of this invention further comprises water, glucose syrup, vitamin C, and sodium citrate. This invention, by mixing the composition containing banana fruit extract with glucose syrup, vitamin C, sodium citrate, and water, does not affect the active function of the composition, and is more suitable for general consumption. Long-term use shows stable effects and no side effects. The water used in this invention is one or more of spring water, small molecule water, purified water, and deionized water, preferably spring water. The beverage obtained by adding the aforementioned composition to this invention has the characteristics of alkalinity, negative potential, and small molecule active ingredients.

[0025] The present invention also provides the use of the composition or the composition obtained by the preparation method or the beverage in the preparation of sleep-improving products.

[0026] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0027] In the following embodiments, the yeast is from Ningbo Mingzhou Biotechnology Co., Ltd., with the serial number BMZ126199; the Lactobacillus plantarum YLP-326 is from Minsheng Zhongke Jiayi (Shandong) Biotechnology Co., Ltd.; the Lactobacillus fermentum is from Ningbo Mingzhou Biotechnology Co., Ltd., with the serial number BMZ134605; and the Streptococcus thermophilus is from Ningbo Mingzhou Biotechnology Co., Ltd., with the serial number BMZ124652.

[0028] Unless otherwise specified, the following embodiments are all conventional methods.

[0029] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0030] Example 1 A composition containing banana fruit extract is made from the following raw materials in parts by weight: 55 parts banana fruit extract, 35 parts apple blueberry extract, 25 parts cherry extract and 35 parts sweet potato leaf extract.

[0031] A method for preparing a composition containing banana fruit extract, comprising the following steps: (1) Select fresh bananas with a maturity of 70-80%, remove the peel and stem, wash them clean and cut them into 0.5-1cm thick pieces. Mix the banana pieces with 75v / v% ethanol at a mass-volume ratio of 1g:25mL, and extract twice by ultrasonication at 55℃ and 250W for 40min each time. Combine the extracts; centrifuge at 2000rpm for 1min, collect the centrifuged liquid, and concentrate it to a soluble fixed content of 10% to obtain banana fruit extract.

[0032] (2) Select ripe fresh apples and blueberries, wash them clean, mix the apples and blueberries at a mass ratio of 1:1, then add water five times the total weight of the apples and blueberries and crush into a pulp; then add pectinase and cellulase according to a total enzyme activity of 250 U / g of apples and blueberries and an enzyme activity ratio of pectinase to cellulase of 1:1, enzymatically hydrolyze at 45℃ for 8 hours to inactivate the enzymes; then add pectinase and cellulase according to a total viable count of 1×10 8 Add cfu / mL enzymatic hydrolysate to yeast and Lactobacillus plantarum YLP-326 (the ratio of live yeast to Lactobacillus plantarum YLP-326 is 2:1), ferment at 32℃ for 40 h, centrifuge at 2500 rpm for 2 min, collect the centrifuged liquid, and concentrate it to a soluble fixed content of 10% to obtain apple blueberry extract.

[0033] (3) Select fresh and plump cherries, wash them and remove the pits; crush the cherries and put them into a supercritical extraction device. Add 95v / v% ethanol according to the mass-volume ratio of cherries to ethanol of 1g:5.5mL. Control the extraction pressure of 19.5MPa and the extraction temperature of 40℃, and continue extraction for 120min. The CO2 flow rate is 16kg / h to allow the active ingredients in the cherries to dissolve fully. After the extraction is completed, freeze dry the extract to finally obtain cherry extract with a water content of 3%.

[0034] (4) After washing the fresh sweet potato leaves, dry them with hot air at 45℃ until the moisture content is ≤10%, and then pulverize them through a 50-mesh sieve to obtain sweet potato leaf powder. Mix the sweet potato leaf powder with water at a mass ratio of 1:20, and extract twice with ultrasound at 45℃ and 300W for 50 minutes each time. Combine the ultrasonic extracts. In the ultrasonic extract, add bacteria according to a total viable count of 6×10⁻⁶. 7 Add cfu / mL ultrasonic extract to Lactobacillus fermentum and Streptococcus thermophilus (live count ratio of Lactobacillus fermentum to Streptococcus thermophilus is 1:2), ferment at 30℃ for 48h, centrifuge at 3000rpm for 1.5min, collect the centrifuged liquid, concentrate to a soluble fixed content of 10%, and obtain sweet potato leaf extract.

[0035] (5) Mix the above-prepared raw material extracts evenly according to the weight parts to obtain a composition containing banana fruit extract.

[0036] A beverage containing banana fruit extract is made from the following ingredients in parts by weight: 20 parts of the above-prepared composition containing banana fruit extract, 3 parts of glucose syrup, 1.5 parts of vitamin C, 0.5 parts of sodium citrate, and 75 parts of spring water.

[0037] The preparation of a beverage containing banana fruit extract involves the following steps: mixing the above raw materials evenly according to their weight proportions, sterilizing, and packaging.

[0038] Example 2 A composition containing banana fruit extract is made from the following raw materials in parts by weight: 50 parts banana fruit extract, 30 parts apple blueberry extract, 20 parts cherry extract and 30 parts sweet potato leaf extract.

[0039] A method for preparing a composition containing banana fruit extract, comprising the following steps: (1) Select fresh bananas with a maturity of 70-80%, remove the peel and stem, wash them clean and cut them into 0.5-1cm thick pieces. Mix the banana pieces with 70v / v% ethanol at a mass-volume ratio of 1g:20mL, and extract them three times under ultrasonic conditions at 50℃ and 200W, with each extraction time being 35min. Combine the extracts; centrifuge at 1500rpm for 2min, collect the centrifuged liquid, and concentrate it until the soluble fixed content is 8% to obtain banana fruit extract.

[0040] (2) Select ripe fresh apples and blueberries, wash them clean, mix the apples and blueberries at a mass ratio of 1:2, then add water three times the total weight of the apples and blueberries and crush into a pulp; then add pectinase and cellulase according to a total enzyme activity of 200 U / g of apples and blueberries and an enzyme activity ratio of pectinase to cellulase of 1:2, enzymatically hydrolyze at 48℃ for 5 hours to inactivate the enzymes; then add pectinase and cellulase according to a total viable count of 0.5 × 10⁻⁶. 8 Add cfu / mL enzymatic hydrolysate to yeast and Lactobacillus plantarum YLP-326 (the ratio of live yeast to Lactobacillus plantarum YLP-326 is 2:1), ferment at 30℃ for 36 h, centrifuge at 2000 rpm for 1 min, collect the centrifuged liquid, and concentrate it to a soluble fixed content of 8% to obtain apple blueberry extract.

[0041] (3) Select fresh and plump cherries, wash them and remove the pits; crush the cherries and put them into a supercritical extraction device. Add ethanol with a volume fraction of 90v / v% according to the mass-volume ratio of cherries to ethanol of 1g:5mL. Control the extraction pressure of 19MPa and the extraction temperature of 38℃, and continue extraction for 110min. The CO2 flow rate is 12kg / h to allow the active ingredients in the cherries to be fully dissolved. After the extraction is completed, freeze dry the extract to finally obtain cherry extract with a water content of 2%.

[0042] (4) After washing the fresh sweet potato leaves, dry them with hot air at 40℃ until the moisture content is ≤10%, then pulverize them through a 40-mesh sieve to obtain sweet potato leaf powder; mix the sweet potato leaf powder with water at a mass ratio of 1:15, and extract them three times with ultrasound at 40℃ and 250W, each extraction time being 45min, and combine the ultrasonic extracts; add the total viable bacteria count to the ultrasonic extracts according to the following ratio: 4×10 7 Add cfu / mL ultrasonic extract to Lactobacillus fermentum and Streptococcus thermophilus (live count ratio of Lactobacillus fermentum to Streptococcus thermophilus is 1:1), ferment at 28℃ for 36 h, centrifuge at 2500 rpm for 2 min, collect the centrifuged liquid, concentrate to a soluble fixed content of 8%, and obtain sweet potato leaf extract.

[0043] (5) Mix the above-prepared raw material extracts evenly according to the weight parts to obtain a composition containing banana fruit extract.

[0044] The preparation of a beverage containing banana fruit extract is the same as in Example 1.

[0045] Example 3 A composition containing banana fruit extract is made from the following raw materials in parts by weight: 60 parts banana fruit extract, 40 parts apple blueberry extract, 30 parts cherry extract and 40 parts sweet potato leaf extract.

[0046] A method for preparing a composition containing banana fruit extract, comprising the following steps: (1) Select fresh bananas with a maturity of 70-80%, remove the peel and stem, wash them clean and cut them into 0.5-1cm thick pieces. Mix the banana pieces with 80v / v% ethanol at a mass-volume ratio of 1g:30mL, and extract twice with ultrasound at 60℃ and 150W for 35min each time. Combine the extracts; centrifuge at 3000rpm for 2min, collect the centrifuged liquid, and concentrate it to a soluble fixed content of 12% to obtain banana fruit extract.

[0047] (2) Select ripe, fresh apples and blueberries, and wash them thoroughly. Mix apples and blueberries at a mass ratio of 1:0.5, then add water at a ratio of 8 times the total weight of apples and blueberries and pulverize into a pulp; then add pectinase and cellulase at a total enzyme activity of 300 U / g of apples and blueberries and an enzyme activity ratio of pectinase to cellulase of 1:1.5, and enzymatically hydrolyze at 40℃ for 10 hours to inactivate the enzymes; then add pectinase and cellulase at a total viable count of 1.5 × 10⁻⁶. 8 Add cfu / mL enzymatic hydrolysate to yeast and Lactobacillus plantarum YLP-326 (live cell ratio of yeast to Lactobacillus plantarum YLP-326 is 1:1), ferment at 35℃ for 48h, centrifuge at 3000rpm for 2min, collect the centrifuged liquid, concentrate to a soluble fixed content of 12%, and obtain apple blueberry extract.

[0048] (3) Select fresh and plump cherries, wash them and remove the pits; crush the cherries and put them into a supercritical extraction device. Add ethanol with a volume fraction of 98v / v% according to the mass-volume ratio of cherries to ethanol of 1g:6.5mL. Control the extraction pressure of 20MPa and the extraction temperature of 42℃, and continue extraction for 140min. The CO2 flow rate is 18kg / h to allow the active ingredients in the cherries to be fully dissolved. After the extraction is completed, freeze dry the extract to finally obtain cherry extract with a water content of 4%.

[0049] (4) After washing the fresh sweet potato leaves, dry them with hot air at 50℃ until the moisture content is ≤10%, and then pulverize them through a 30-mesh sieve to obtain sweet potato leaf powder. Mix the sweet potato leaf powder with water at a mass ratio of 1:25, and extract twice with ultrasound at 50℃ and 350W for 40 minutes each time. Combine the ultrasonic extracts. In the ultrasonic extract, add bacteria according to the total viable count of 6.5×10⁻⁶. 7 The cfu / mL ultrasonic extract was added to Lactobacillus fermentum and Streptococcus thermophilus (mass ratio of Lactobacillus fermentum to Streptococcus thermophilus was 1:2), fermented at 32℃ for 24 h, centrifuged at 3500 rpm for 2 min, the centrifuged liquid was collected, and concentrated to a soluble fixed content of 12% to obtain sweet potato leaf extract.

[0050] (5) Mix the above-prepared raw material extracts evenly according to the weight parts to obtain a composition containing banana fruit extract.

[0051] The preparation of a beverage containing banana fruit extract is the same as in Example 1.

[0052] Comparative Example 1 The sweet potato leaf extract was replaced with banana fruit extract, and the preparation method of the sweet potato leaf extract was removed. Everything else was the same as in Example 1.

[0053] Comparative Example 2 The apple blueberry extract was replaced with banana fruit extract, and the preparation method of the apple blueberry extract was removed. Everything else was the same as in Example 1.

[0054] Comparative Example 3 The banana fruit extract was prepared by replacing ethanol with water; the enzymatic hydrolysis step in the preparation of the apple and blueberry extract was removed; the rest were the same as in Example 1.

[0055] Comparative Example 4 The fermenting lactobacillus and thermophilic streptococcus used in the preparation of sweet potato leaf extract were replaced with yeast and Lactobacillus plantarum YLP-326 used in the preparation of apple blueberry extract, and the remaining steps were the same as in Example 1.

[0056] Comparative Example 5 The method for preparing cherry extract is replaced by: putting the processed cherries into a juicer to extract the juice, then filtering it through a fine mesh sieve to remove fruit pulp residue and collecting the pure cherry juice. The remaining steps are the same as in Example 1.

[0057] Experimental Example 1 1. Experimental animals: SPF-grade mice were acclimatized for 7 days before the experiment. Environmental conditions: temperature 22-25℃, humidity 50%-60%, 12h light / 12h dark cycle (light time 7:00-19:00), free access to food and water.

[0058] 2. Grouping, modeling, and drug administration (1) The adapted mice were randomly divided into 10 groups: normal control group, model group, Examples 1-3 (compositions prepared in each group) and Comparative Examples 1-5 (compositions prepared in each group), with 10 mice in each group.

[0059] (2) For three consecutive days, mice were injected intraperitoneally with a suspension of p-chlorophenylalanine (concentration of 0.03 g / mL) at a dose of 300 mg / kg. Successful modeling was indicated when mice exhibited restlessness, agitation, and increased daytime activity. The normal control group received an equal volume of physiological saline intraperitoneally, while the other groups were modeled using the same method. After successful modeling, mice were administered the drug via gavage starting the following morning for seven consecutive days, with a gavage volume of 2 mL / 100 g. The normal control group and the model group received an equal volume of distilled water via gavage.

[0060] Mouse spontaneous activity experiment: 40 minutes after the last administration, mice in each group were placed in the test box of the spontaneous activity recorder. After acclimatization for 5 minutes, the number of spontaneous activities (number of times the forelimbs were raised) of the mice within 5 minutes was recorded.

[0061] Table 1 Results of spontaneous activity experiments in each group of mice

[0062] Table 1 shows that the model group exhibited a significant increase in activity levels compared to the normal control group, indicating successful model establishment. Compared to the model group, both Examples 1-3 and Comparative Examples 1-5 reduced mouse activity levels, but the reduction in Examples 1-3 was significantly greater than that in Comparative Examples 1-5, and essentially approached the level of the normal control group. This indicates that the four extracts (banana fruit, apple blueberry, cherry, and sweet potato leaf) have a synergistic effect in improving sleep, and the absence of any one of them would reduce the overall effect. It also demonstrates that the combination of extracts prepared using the specific method of this invention has a synergistic effect, significantly reducing the number of spontaneous activities in the model group mice and synergistically improving sleep.

[0063] (3) Following the grouping and modeling methods described above, 40 minutes after the last administration, each group of mice was intraperitoneally injected with 0.01 mL / g of sodium pentobarbital (30 mg / kg). 30 minutes after administration, the disappearance and recovery times of the righting reflex were recorded for each group of mice. Sleep duration (from the disappearance of the righting reflex to its reappearance) was calculated, and the results are shown in Table 2. A longer sleep duration indicates a more significant improvement in sleep maintenance.

[0064] Table 2 Sleep duration of each group

[0065] As shown in Table 2, compared with the model group, Examples 1-3 and Comparative Examples 1-5 all showed a trend of prolonging the sleep duration of mice. However, Examples 1-3 were significantly better than Comparative Examples 1-5, and basically reached the normal level. This further demonstrates that the extract combinations obtained by the extraction methods of each raw material specified in this invention have synergistic effects, with the active ingredients in each extract complementing and interacting with each other, resulting in a significant improvement in mouse sleep.

[0066] Experimental Example 2 The beverages obtained in Examples 1-3 were subjected to acute oral toxicity tests, and the test methods were in accordance with the National Food Safety Standard for Acute Oral Toxicity Tests GB 15193.3-2014.

[0067] SPF-grade mice meeting national standards and relevant regulations, weighing 18.0g-20.0g, were randomly divided into 3 groups of 10 mice each (half male and half female). Mice were fasted for 6 hours before the experiment but had free access to water. Each group was then given a single dose of the experimental solution at 10000mg / kg body weight (50g of the prepared beverage from the previous example was mixed with ultrapure water to a final volume of 100mL). One hour later, mice were given their normal diet. The mice were observed for 14 days, and symptoms of poisoning and mortality were recorded. Live animals were weighed weekly during the observation period. At the end of the observation period, surviving animals were humanely euthanized and autopsied. The results are shown in Table 3.

[0068] Table 3 Results of acute oral toxicity test of beverages in mice

[0069] The experimental results showed that no obvious symptoms of poisoning or death were observed in any of the test animals during the observation period, and no abnormalities were found in the gross anatomical examination of the experimental animals. Therefore, the acute oral toxicity LD50 in mice is [not specified]. 50 >10000mg / kg body weight, classified as a non-toxic substance.

[0070] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A composition containing banana fruit extract, characterized in that, It includes the following ingredients by weight: 45-65 parts banana fruit extract, 25-45 parts apple blueberry extract, 15-35 parts cherry extract and 25-45 parts sweet potato leaf extract.

2. The method for preparing the composition according to claim 1, characterized in that, Includes the following steps: Banana fruit was mixed with ethanol, sonicated, centrifuged, and the liquid was collected and concentrated to obtain banana fruit extract. Apples and blueberries are crushed into a pulp, and pectinase and cellulase are added for enzymatic hydrolysis to inactivate the enzymes. Then, yeast and Lactobacillus plantarum YLP-326 are added for fermentation. The centrifuged liquid is collected, concentrated, and apple-blueberry extract is obtained. Cherries were crushed, and ethanol was added for supercritical CO2 extraction. The extract was then freeze-dried to obtain cherry extract. Sweet potato leaf powder was ultrasonically treated with water, then fermented with Lactobacillus fermentum and Streptococcus thermophilus. The centrifuged liquid was collected, concentrated, and sweet potato leaf extract was obtained.

3. The preparation method according to claim 2, characterized in that, The temperature of the mixture of banana fruit and ethanol is 50-60℃, the power is 150-250W, the time is 30-50min, and the number of times is 1-3.

4. The preparation method according to claim 2, characterized in that, The ratio of pectinase to cellulase activity is 1:0.5-1.5; the total enzyme activity of pectinase and cellulase is 200-300 U / g of the total weight of apples and blueberries; the enzymatic hydrolysis temperature is 40-50℃ and the time is 3-10h.

5. The preparation method according to claim 2, characterized in that, The ratio of viable cell number of the yeast and Lactobacillus plantarum YLP-326 is 1-3:1; the total viable cell number of the yeast and Lactobacillus plantarum YLP-326 is 0.5-1.5×10 8 cfu / mL of the enzymatic hydrolysate; the fermentation temperature is 27-35℃, and the fermentation time is 24-48h.

6. The preparation method according to claim 2, characterized in that, The mass-to-volume ratio of cherries to ethanol is 1g:4-7mL; the extraction pressure is 19-20MPa, the temperature is 38-42℃, the time is 100-140min, and the CO2 flow rate is 12-18kg / h.

7. The preparation method according to claim 2, characterized in that, The ratio of viable number of the Lactobacillus fermentum to the Streptococcus thermophilus is 1:1-3; the total viable number of the Lactobacillus fermentum and the Streptococcus thermophilus is 3.5-6.5×10 7 cfu / mL ultrasonic extraction solution; the fermentation temperature is 27-35℃, and the fermentation time is 24-48h.

8. A beverage containing banana fruit extract, characterized in that, The composition includes the composition of claim 1 or the composition obtained by the preparation method of any one of claims 2-7.

9. The beverage as described in claim 8, characterized in that, It also includes water, glucose syrup, vitamin C, and sodium citrate.

10. The use of the composition of claim 1, the composition obtained by the preparation method of any one of claims 2-7, or the beverage of claim 8 in the preparation of a sleep-improving product.

Images