A fermented composition of jujube and its use in anti-fatigue and sleep improvement

Abstract

This invention discloses a jujube fermentation composition and its use in anti-fatigue and sleep improvement, belonging to the field of health products. It comprises the following components by weight: 10-30 parts of jujube pit water extract and 60-85 parts of jujube pulp. This invention fully utilizes jujube pit resources, turning waste into treasure, improving the comprehensive utilization value of jujubes, and reducing production costs. Through fermentation with *Lactobacillus plantarum* and *Lactobacillus acidophilus*, this invention transforms the active ingredients in jujubes, improving bioavailability and enhancing product functionality. The jujube fermentation composition of this invention is of natural origin, highly safe, and non-addictive, making it suitable for long-term use. The preparation process of this invention is simple, easy to industrialize, and the product quality is controllable. This invention can simultaneously exert the dual effects of anti-fatigue and sleep improvement, meeting the multiple health needs of modern people.

Description

Technical Field

[0001] This invention belongs to the field of health products, specifically a jujube fermentation composition and its use in anti-fatigue and sleep improvement. Background Technology

[0002] In today's fast-paced lifestyle, high work pressure and irregular schedules have led to increasingly common fatigue and sleep disorders. Statistics show that about one-third of the global population experiences fatigue to varying degrees, while the prevalence of sleep disorders is as high as 40%. Chronic fatigue and sleep deprivation not only affect work efficiency and quality of life, but also lead to weakened immunity, endocrine disorders, and an increased risk of cardiovascular disease, depression, and many other illnesses.

[0003] Currently, the main drugs used clinically to improve fatigue and sleep disorders include central nervous system stimulants and sedative-hypnotics. While central nervous system stimulants such as caffeine can temporarily relieve fatigue, long-term use can lead to dependence and may cause adverse reactions such as palpitations, anxiety, and insomnia. Sedative-hypnotics such as benzodiazepines can improve sleep, but they have problems such as addiction, residual effects the next day, and memory impairment. Therefore, it is of great significance to develop safe, effective, and non-addictive natural products for combating fatigue and improving sleep.

[0004] Jujube is a plant belonging to the genus Ziziphus of the family Rhamnaceae. It is a traditional Chinese food and medicine with the same origin, and has the effects of replenishing qi and blood and calming the mind. Modern research shows that jujube contains a variety of active ingredients such as polysaccharides, flavonoids, saponins, and cyclic adenosine monophosphate (cAMP), which have antioxidant, immunomodulatory, and neuroprotective effects. However, traditional jujube processing methods have problems such as low extraction rate of active ingredients and poor bioavailability. Summary of the Invention

[0005] This invention provides a jujube fermentation composition and its use in anti-fatigue and sleep improvement, in order to overcome the deficiencies in the prior art.

[0006] This invention is achieved through the following technical solution: A jujube fermentation composition comprises the following substances in parts by weight: 10-30 parts of jujube pit water extract and 60-85 parts of jujube pulp.

[0007] The jujube fermentation composition described above, wherein the preparation operation of the jujube pit water extract is as follows: take dried jujube pits, crush them, add 8 to 12 times the weight of purified water, extract at 80 to 95°C for 2 to 3 hours, filter, repeat the above extraction operation 1 to 2 times with the filter residue, combine the filtrates, concentrate under reduced pressure until the solid content is 10% to 20%, and thus obtain the jujube pit water extract.

[0008] In the jujube fermentation composition described above, the jujube pits are crushed and passed through a 40-60 mesh screen for later use. If there are residues on the screen, they are either crushed again or recycled and added back during the next crushing.

[0009] The jujube fermentation composition described above, wherein the preparation of the jujube pulp is as follows: select ripe and plump jujubes, wash them, blanch them in boiling water for 2-3 minutes, peel them, remove the pits, add 2-4 times the weight of purified water, and blend them to obtain the jujube pulp.

[0010] In the jujube fermentation composition described above, the pulping operation is performed using a colloid mill for homogenization, and the rotation speed of the colloid mill is 5000-8000 r / min.

[0011] The jujube fermentation composition described above is prepared by the following steps: Step 1: Mix the jujube pit water extract with jujube pulp according to the specified ratio, adjust the pH to 6.0-6.5, and sterilize. Step 2: After cooling, inoculate with fermentation starter cultures. The ratio of viable Lactobacillus plantarum to Lactobacillus acidophilus should be 10–5:1, and the total inoculation amount should be 1 × 10⁻⁶. 6 ~1×10 8 CFU / mL; Step 3: Ferment at 35-40℃ for 24-72 hours, controlling the pH to be no lower than 4.0 during fermentation; Step 4: After fermentation, the mixture is sterilized and cooled to obtain the jujube fermentation composition.

[0012] The jujube fermentation composition described above includes Lactobacillus plantarum and Lactobacillus acidophilus as fermentation strains.

[0013] In the jujube fermentation composition described above, the ratio of viable Lactobacillus plantarum to Lactobacillus acidophilus is 10–5:1, and the total inoculum is 1 × 10⁻⁶. 6 ~1×10 8 CFU / mL.

[0014] The use of a jujube fermentation composition in anti-fatigue and sleep improvement, wherein the jujube fermentation composition can play the role of anti-fatigue and sleep improvement, and the daily intake of the jujube fermentation composition for adults is 200-500g, the daily intake for children is 100-200g, and the daily intake for infants is 10-50g.

[0015] The above-mentioned use of a jujube fermentation composition in anti-fatigue and sleep improvement, the jujube fermentation composition can be further processed into oral liquids, beverages and jellies to enhance flavor and texture.

[0016] The advantages of this invention are: It fully utilizes jujube kernel resources, turning waste into treasure, improving the comprehensive utilization value of jujubes, and reducing production costs; through fermentation with *Lactobacillus plantarum* and *Lactobacillus acidophilus*, it transforms the active ingredients in jujubes, improving bioavailability and enhancing product functionality; the jujube fermentation composition of this invention is of natural origin, highly safe, non-drug-dependent, and suitable for long-term use; the preparation process of this invention is simple, easy to industrialize, and the product quality is controllable; this invention can simultaneously exert the dual effects of anti-fatigue and sleep improvement, meeting the multiple health needs of modern people. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0018] Figure 1 This is a schematic diagram comparing the effect of the jujube fermentation composition of the present invention with other experimental groups on the weight-bearing swimming time of mice; Figure 2 This is a schematic diagram comparing the effects of the jujube fermentation composition of the present invention with other experimental groups on the liver glycogen and muscle glycogen content in mice; Figure 3 This is a schematic diagram comparing the effects of the jujube fermentation composition of the present invention with other experimental groups on the sleep latency of mice; Figure 4 This is a schematic diagram comparing the effects of the jujube fermentation composition of the present invention with other experimental groups on the sleep time of mice. Detailed Implementation

[0019] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, 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, 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.

[0020] A jujube fermentation composition comprises the following substances in parts by weight: 10-30 parts of jujube pit water extract and 60-85 parts of jujube pulp.

[0021] Preferably, the preparation operation of the jujube kernel water extract in this embodiment is as follows: take dried jujube kernels, crush them, add 8 to 12 times the weight of purified water, extract at 80 to 95°C for 2 to 3 hours, filter, repeat the above extraction operation 1 to 2 times with the filter residue, combine the filtrates, and concentrate under reduced pressure until the solid content is 10% to 20% to obtain the jujube kernel water extract.

[0022] Preferably, the jujube pits described in this embodiment are crushed and passed through a 40-60 mesh for later use. If there are residues on the sieve, they are crushed again or recycled and added back during the next crushing.

[0023] Preferably, the preparation operation of the jujube pulp in this embodiment is as follows: select ripe and plump jujubes, wash them, blanch them in boiling water for 2 to 3 minutes, peel them, remove the pits, add 2 to 4 times the weight of purified water, and blend them to obtain jujube pulp.

[0024] Preferably, the pulping operation in this embodiment employs homogenization using a colloid mill, with the mill speed being 5000–8000 r / min. Preferably, the preparation method in this embodiment includes the following steps: Step 1: Mix the jujube pit water extract with jujube pulp according to the specified ratio, adjust the pH to 6.0-6.5, and sterilize. Step 2: Inoculate with fermentation starter culture after cooling; Step 3: Ferment at 35-40℃ for 24-72 hours, controlling the pH to be no lower than 4.0 during fermentation; Step 4: After fermentation, the mixture is sterilized and cooled to obtain the jujube fermentation composition.

[0025] Preferably, the fermentation strains described in this embodiment include Lactobacillus plantarum and Lactobacillus acidophilus.

[0026] Preferably, in this embodiment, the live bacteria ratio of *Lactobacillus plantarum* and *Lactobacillus acidophilus* is 10–5:1, and the total inoculum size is 1 × 10⁻⁶. 6 ~1×10 8 CFU / mL.

[0027] The use of a jujube fermentation composition in anti-fatigue and sleep improvement, wherein the jujube fermentation composition can play the role of anti-fatigue and sleep improvement, and the daily intake of the jujube fermentation composition for adults is 200-500g, the daily intake for children is 100-200g, and the daily intake for infants is 10-50g.

[0028] Preferably, the jujube fermentation composition described in this embodiment can be further processed into oral liquids, beverages, and jellies to enhance flavor and texture.

[0029] Example 1: Preparation of Jujube Kernel Water Extract Take 10 kg of dried jujube pits, crush them through a 40-mesh sieve, add 100 kg of purified water, and extract at 90℃ for 2.5 hours, then filter through a 200-mesh sieve. Add 80 kg of purified water to the residue, extract at 90℃ for 2 hours, and filter. Combine the two filtrates, concentrate under reduced pressure until the solid content is 15%, and obtain approximately 12 kg of jujube pit aqueous extract.

[0030] Example 2: Preparation of Jujube Pulp Select 50kg of ripe and plump Xinjiang Hotan jujubes, wash and remove the pits, add 150kg of purified water, pulp them using a pulping machine, and then homogenize them using a colloid mill to obtain about 180kg of jujube pulp.

[0031] Example 3: Preparation of Jujube Fermentation Composition Take 20 kg of jujube kernel water extract prepared in Example 1 and 75 kg of jujube pulp prepared in Example 2, mix and stir evenly, adjust the pH to 6.2, and sterilize at 115℃ for 20 minutes. Cool to 37℃ and inoculate with Lactobacillus plantarum (1×10⁻⁶). 7 CFU / mL) and Lactobacillus acidophilus (2×10⁻⁶ CFU / mL) 6 The mixture (CFU / mL) was fermented at 37℃ for 48 hours. After fermentation, it was sterilized at 80℃ for 15 minutes and then cooled to obtain the jujube fermentation composition.

[0032] Example 4: Jujube Fermented Oral Liquid Take the jujube fermentation composition prepared in Example 3, add 5% honey for flavoring, homogenize, and fill (10mL / vial) to obtain jujube fermentation oral liquid.

[0033] Example 5: Jujube Fermented Beverage The jujube fermentation composition prepared in Example 3 was diluted with water to a soluble solids content of 8%, citric acid and honey were added for flavoring, and the mixture was then sterilized by UHT and bottled to obtain the jujube fermentation beverage.

[0034] Verification test I. Evaluation of Anti-fatigue Effect Laboratory animals: SPF-grade male ICR mice, weighing 18–22 g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0035] Test samples: Jujube fermentation composition prepared in Example 3 (hereinafter referred to as fermentation composition), unfermented jujube pit water extract + jujube pulp mixture (hereinafter referred to as unfermented group), pure jujube pulp (hereinafter referred to as pulp group), positive control: Rhodiola rosea extract.

[0036] Experimental instruments: Mouse swimming apparatus: glass container with a diameter of 18cm and a height of 40cm, electronic balance, centrifuge, enzyme-linked immunosorbent assay (ELISA) reader, biochemical analyzer, etc.

[0037] Grouping and Administration: After 7 days of acclimatization, mice were randomly divided into 6 groups of 12 mice each: model group, original pulp group (506 mg / kg), unfermented group (506 mg / kg), low-dose fermented composition group (253 mg / kg), high-dose fermented composition group (506 mg / kg), and positive control group (506 mg / kg). All mice were administered the drug by gavage once daily for 14 days. The model group was given distilled water by gavage, while the positive control group was given Rhodiola rosea extract by gavage. A weighted swimming test was conducted 30 minutes after the last administration.

[0038] Indicator 1: Swimming time with weight: Group Dosage (mg / kg) Swimming time with added weight (min) Elongation rate (%) Significant difference Model group - 10.8±1.5 - - Original pulp group 506 13.3±1.8 23.1 * Unfermented group 506 15.7±2.1 45.4 ** Low-dose group of fermentation composition 253 16.2±1.9 50.0 ** High-dose group of fermentation composition 506 19.5±2.4 80.6 **△△ Positive control group 200 17.8±2.2 64.8 ** Note: Compared with the model group, *P<0.05, **P<0.01; compared with the unfermented group, △△P<0.01. (From Table 1 and...) Figure 1 The experimental results showed that, compared with the model group, the weight-bearing swimming time of mice in each drug-treated group was significantly prolonged (P<0.05 or P<0.01). Among them, the high-dose group of fermented composition had the most significant effect, with the weight-bearing swimming time increasing from 10.8 minutes to 19.5 minutes, an extension rate of 80.6%, which was significantly better than the unfermented group (extension rate 45.4%) and the original pulp group (extension rate 23.1%). This indicates that the jujube fermented composition of the present invention has a significant anti-fatigue effect, and its activity is significantly enhanced after fermentation treatment.

[0039] Indicator 2: Liver glycogen and muscle glycogen measurement: Group Liver glycogen (mg / g) Muscle glycogen (mg / g) Model group 8.2±1.3 1.5±0.3 Original pulp group 9.8±1.5 1.8±0.4 Unfermented group 11.5±1.8* 2.1±0.4* Low-dose group of fermentation composition 12.8±2.1** 2.3±0.5** High-dose group of fermentation composition 14.1±2.3**△ 2.4±0.5**△ Positive control group 13.2±2.0** 2.2±0.4* Note: Compared with the model group, *P<0.05, **P<0.01; compared with the unfermented group, △P<0.05.

[0040] From Table 2 and Figure 2 It was found that, compared with the model group, the high-dose fermented composition group showed significantly increased liver and muscle glycogen content in mice (P<0.01), increasing by 72.3% and 58.6% respectively, with significantly better effects than the unfermented group and the original pulp group. Glycogen is an important energy source during exercise, and increased glycogen reserves help improve exercise endurance and delay fatigue.

[0041] Detection indicator 3: Serum biochemical indicators Blood was collected from the orbital cavity of mice, serum was separated, and the levels of lactate (LA), blood urea nitrogen (BUN), lactate dehydrogenase (LDH), and creatine kinase (CK) were measured. The results are shown in Table 3.

[0042] Group Lactic acid (nmol / L) Blood urea nitrogen (nmol / L) LDH(U / L) CK(U / L) Model group 7.8±1.2 12.5±2.1 1250±180 485±75 Original pulp group 6.5±1.0* 10.1±1.6* 1120±165* 421±63* Unfermented group 6.1±0.9** 9.3±1.5** 1045±152** 386±59** Low-dose group of fermentation composition 5.9±0.9** 9.8±1.5* 980±150** 375±58* High-dose group of fermentation composition 4.5±0.8**△ 7.2±1.2**△ 820±120**△ 298±48**△ Positive control group 5.2±0.9** 8.5±1.4** 890±140** 325±52** Note: Compared with the model group, *P<0.05, **P<0.01; compared with the unfermented group, △P<0.05.

[0043] Table 3 shows that, compared with the model group, the serum lactate, blood urea nitrogen, LDH, and CK levels in the high-dose fermentation composition group were significantly reduced (P<0.01). Serum lactate and blood urea nitrogen are important indicators of fatigue level; their reduced levels indicate a decrease in the accumulation of metabolic products and a reduction in fatigue. LDH and CK are marker enzymes of myocardial and skeletal muscle damage; their reduced activity indicates that the fermentation composition has a protective effect on muscles.

[0044] Detection index 4: Oxidative stress index Liver tissue was collected, homogenized, and the malondialdehyde (MDA) content and the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were determined. The results are shown in Table 4.

[0045] Group MDA (nmol / mg) SOD (U / mg) GSH = Px(u / mg) Model group 8.5±1.2 125±18 28±5 Original pulp group 7.2±1.0* 142±20* 33±6* Unfermented group 6.9±0.9** 150±21** 36±6** Low-dose group of fermentation composition 6.8±0.9* 158±22** 38±6** High-dose group of fermentation composition 5.2±0.8**△ 185±25**△ 48±7**△ Positive control group 5.8±0.9** 172±24** 42±6** Note: Compared with the model group, *P<0.05, **P<0.01; compared with the unfermented group, △P<0.05.

[0046] As shown in Table 4, compared with the model group, the MDA content in the liver of mice in the high-dose group of fermented composition was significantly reduced (P<0.01), and the SOD and GSH-Px activities were significantly increased (P<0.01), indicating that the jujube fermented composition of the present invention has significant antioxidant effects and can reduce exercise-induced oxidative stress damage.

[0047] II. Evaluation of Sleep-Aid Effect Experimental animals: SPF-grade ICR mice, male, weighing 18–22g.

[0048] Test samples: Jujube fermentation composition prepared in Example 3 (hereinafter referred to as fermentation composition), unfermented jujube pit water extract + jujube pulp mixture (hereinafter referred to as unfermented group), simple jujube pulp (hereinafter referred to as pulp group), positive control: diazepam.

[0049] Grouping and administration: After 7 days of acclimatization, mice were randomly divided into 6 groups of 12 mice each: model group, original pulp group (506 mg / kg), unfermented group (506 mg / kg), low-dose fermented composition group (253 mg / kg), high-dose fermented composition group (506 mg / kg), and positive control group (diazepam, 2 mg / kg. Diazepam has a strong effect, and the above dose can achieve the corresponding sleep-inducing effect. Moreover, high doses can easily lead to the death of mice. Therefore, the different dosages here do not affect the comparison of effects. Each group of mice was administered the drug by gavage once a day for 14 consecutive days.

[0050] Synergistic pentobarbital sleep experiment: Sixty minutes after the last oral administration, mice in each group were intraperitoneally injected with sodium pentobarbital (40 mg / kg). Sleep latency (time from injection of sodium pentobarbital to the disappearance of the righting reflex) and sleep duration (time from the disappearance of the righting reflex to its recovery) were recorded. The results are shown in Table 5 and... Figure 3 As shown.

[0051] Group Dosage (mg / kg) Sleep latency (min) Shortening rate (%) Model group - 7.2±0.8 - Original pulp group 506 6.4±0.7 11.1 Unfermented group 506 5.5±0.6* 23.6 Low-dose group of fermentation composition 253 5.0±0.6** 30.6 High-dose group of fermentation composition 506 4.1±0.5**△△ 43.1 Positive control group 2 3.8±0.5** 47.2 Note: Compared with the model group, *P<0.05, **P<0.01; compared with the unfermented group, △△P<0.01.

[0052] From Table 5 and Figure 3 It was found that, compared with the model group, the sleep latency of mice in the high-dose fermentation composition group was significantly shortened (P<0.01), from 7.2 minutes to 4.1 minutes, with a shortening rate of 43.1%, which was significantly better than the unfermented group (shortening rate of 23.6%) and the original pulp group (shortening rate of 11.1%).

[0053] Sleep EEG monitoring experiment: Mice were anesthetized with sodium pentobarbital and had EEG electrodes implanted. After 7 days of recovery, EEG and EMG recordings were performed following drug administration. Sleep-wake cycles were analyzed, including: total sleep time (TST), sleep latency (SOL), slow-wave sleep (SWS) time and proportion, rapid eye movement (REM) sleep time and proportion, and number of awakenings. Results are shown in Tables 6 and 7. Figure 4 As shown.

[0054] Group Dosage (mg / kg) Sleep duration (min) Elongation rate (%) Model group - 92±12 - Original pulp group 506 105±14 14.1 Unfermented group 506 122±15* 32.6 Low-dose group of fermentation composition 253 138±16** 50.0 High-dose group of fermentation composition 506 158±18**△△ 71.7 Positive control group 2 164±20** 78.3 Note: Compared with the model group, *P<0.05, **P<0.01; compared with the unfermented group, △△P<0.01.

[0055] Group TST(min) SWS(min) SWS(%) REM(min) Awakening times Model group 385±42 245±28 63.6 48±8 28±5 Original pulp group 412±45 268±30 65.0 50±8 25±4 Unfermented group 435±47* 282±31* 64.8 51±9 24±4* Low-dose group of fermentation composition 458±48** 298±32** 65.1 52±9 22±4* High-dose group of fermentation composition 522±55**△ 352±38**△ 67.4 55±10 16±3**△ Positive control group 540±60 ∗∗ 365±40 ∗∗ 68.2 58±11 14±2 ∗∗ Note: Compared with the model group, *P<0.05, **P<0.01; compared with the unfermented group, △P<0.05.

[0056] As shown in Table 7 and Figure 1, the total sleep time of mice in the high-dose group of fermented composition was significantly increased (P<0.01), mainly due to the significant prolongation of slow-wave sleep (SWS) time (P<0.01), while the effect on rapid eye movement sleep (REM) was small, and the number of awakenings was significantly reduced (P<0.01), indicating that the sleep quality was improved.

[0057] III. Determination of Neurotransmitter and Neuropeptide Content To explore the mechanism of the sleep-aiding effect of jujube fermentation composition, its effects on neurotransmitters and neuropeptides in the brain were determined.

[0058] Experimental Methods: After 7 days of acclimatization, mice were randomly divided into 6 groups of 12 mice each: blank control group, original plasma group (506 mg / kg), unfermented group (506 mg / kg), low-dose fermentation composition group (253 mg / kg), high-dose fermentation composition group (506 mg / kg), and positive control group (2 mg / kg). Mice in each group were administered the drug once daily by gavage for 14 consecutive days. After the last administration, the mice were sacrificed, and their brains were rapidly harvested. The hypothalamus and cortex were separated, flash-frozen in liquid nitrogen, and stored at -80°C. The levels of γ-aminobutyric acid (GABA), glutamate (Glu), serotonin (5-HT), norepinephrine (NE), and dopamine (DA), as well as the levels of orexin A (Orexin A) and melatonin (MT), were determined according to the kit instructions. The results are shown in Table 8.

[0059] index Blank group Original pulp group Unfermented group Low-dose group of fermentation composition High-dose group of fermentation composition positive group Hypothalamic GABA (μg / g) 125±18 142±20 165±22* 182±24** 198±25** 215±28** Cortical GABA (μg / g) 98±15 110±16 130±18* 141±19** 152±20** 165±23** Glu / GABA ratio 2.85±0.42 2.60±0.38 2.20±0.35* 1.90±0.30** 1.68±0.28** 1.52±0.25** 5-HT (ng / g) 285±38 310±40 350±42* 375±45** 398±48** 420±52** NE(ng / g) 168±22 155±20 140±18* 134±17* 128±18* 115±15** Orexin A (ng / mL) 42±6 38±5 32±4* 30±4** 28±4** 24±3** Melatonin (pg / mL) 85±12 95±14 105±15* 112±16* 118±15* 130±18** Note: Compared with the model group, *P<0.05, **P<0.01; compared with the unfermented group, △P<0.05.

[0060] Table 8 shows that, compared with the model group, the high-dose fermentation composition group of mice showed significantly increased GABA content in the hypothalamus and cortex (P<0.01), significantly decreased Glu / GABA ratio (P<0.01), significantly increased 5-HT content (P<0.01), significantly decreased NE content (P<0.05), significantly decreased orexin A content (P<0.01), and significantly increased melatonin content (P<0.05). These results indicate that GABA in the jujube fermentation composition (derived from the aqueous extract of jujube kernels) works synergistically with other active ingredients produced during fermentation, possibly by regulating the GABAergic and 5-HTergic systems, inhibiting orexin neuronal activity, and promoting melatonin secretion, thereby exerting a sleep-inducing effect.

[0061] IV. Safety Evaluation Acute toxicity test: Forty healthy mice (half male and half female, weighing 22±2g) were selected and fasted for 12 hours. They were then administered the jujube fermentation composition of this invention by gavage at the maximum concentration (100%) and maximum volume (0.4mL / 10g body weight), equivalent to 20g / kg (based on crude drug weight). Fasting continued for another 4 hours after administration, and the mice were observed for 14 days. Results: No abnormal reactions or deaths were observed in any of the mice. This indicates that the jujube fermentation composition of this invention has extremely low acute toxicity, with an LD50 greater than 20g / kg.

[0062] Long-term toxicity testing: Eighty healthy rats, half male and half female, were randomly divided into four groups of 20 rats each. A control group and low-, medium-, and high-dose groups (2.5, 5.0, and 10.0 g / kg, respectively) were included. The rats were administered the drug once daily by gavage for 90 consecutive days. Results: The rats in all groups were in good general condition, with normal weight gain. No abnormalities related to the test substance were observed in hematological, blood biochemistry, organ coefficient, and histopathological examinations, indicating that the jujube fermentation composition of this invention has good safety under long-term administration.

[0063] V. Human Trial Testing Subjects: Sixty volunteers aged 25-55 years with fatigue and / or sleep disorders were recruited. No gender restriction was applied. Exclusion criteria: severe liver and kidney dysfunction, pregnant and lactating women, and those currently taking sedative-hypnotic drugs.

[0064] Experimental Methods: A randomized, double-blind, controlled design was adopted. Volunteers were randomly assigned to an experimental group and a control group, with 30 participants in each group. The experimental group took the jujube fermented oral liquid prepared in Example 4, 10 mL twice daily; the control group took a placebo, with the same dosage and administration. The treatment lasted for 4 weeks.

[0065] index Experimental group (before the experiment) Experimental group (after 4 weeks) Control group (after 4 weeks) FSS score 42.5±6.8 28.3±5.2**△△ 40.2±7.1 PSQI score 14.2±3.5 7.8±2.4**△ 13.5±3.8 Sleep efficiency (%) 78.5±8.2 88.2±6.5*△ 80.1±7.8 Awakening times 6.5±2.1 3.2±1.5**△ 5.8±2.3 Note: Compared with pre-experiment values, *P<0.05, **P<0.01; compared with the control group, △P<0.05, △△P<0.01. As shown in Table 9, after 4 weeks of treatment, the FSS and PSQI scores of the experimental group were significantly lower than before the trial (P<0.01) and significantly lower than those of the control group (P<0.05 or P<0.01). Sleep monitoring showed that the sleep efficiency of the experimental group was significantly improved (P<0.05) and the number of awakenings was significantly reduced (P<0.05).

[0066] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do 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. A jujube fermentation composition, characterized in that: The substance includes the following parts by weight: 10-30 parts of jujube kernel water extract and 60-85 parts of jujube pulp.

2. The jujube fermentation composition according to claim 1, characterized in that: The preparation procedure for the jujube kernel water extract is as follows: take dried jujube kernels, crush them, add 8 to 12 times their weight of purified water, extract at 80 to 95°C for 2 to 3 hours, filter, repeat the above extraction operation 1 to 2 times with the filter residue, combine the filtrates, and concentrate under reduced pressure until the solid content is 10% to 20% to obtain the jujube kernel water extract.

3. The jujube fermentation composition according to claim 2, characterized in that: The jujube pits are crushed and passed through a 40-60 mesh screen for later use. If there are residues on the screen, they are crushed again or recycled and added back during the next crushing.

4. The jujube fermentation composition according to claim 1, characterized in that: The preparation process of the jujube pulp is as follows: Select ripe and plump jujubes, wash them, blanch them in boiling water for 2-3 minutes, peel them, remove the pits, add 2-4 times the weight of purified water, and blend them to obtain the jujube pulp.

5. The jujube fermentation composition according to claim 4, characterized in that: The pulping operation is performed using a colloid mill for homogenization, and the rotation speed of the colloid mill is 5000-8000 r / min.

6. The jujube fermentation composition according to claim 1, characterized in that: Its preparation method includes the following steps: Step 1: Mix the jujube pit water extract with jujube pulp according to the specified ratio, adjust the pH to 6.0-6.5, and sterilize. Step 2: inoculate the fermentation strain after cooling, the ratio of viable cell number of Lactobacillus plantarum and Lactobacillus acidophilus is 10:1, and the total inoculation amount is 1 x 10 6 CFU / mL 8 CFU / mL Step 3: Ferment at 35-40℃ for 24-72 hours, controlling the pH to be no lower than 4.0 during fermentation; Step 4: After fermentation, the mixture is sterilized and cooled to obtain the jujube fermentation composition.

7. The jujube fermentation composition according to claim 6, characterized in that: The fermentation strains include Lactobacillus plantarum and Lactobacillus acidophilus.

8. The jujube fermentation composition according to claim 7, characterized in that: The ratio of viable Lactobacillus plantarum to Lactobacillus acidophilus is 10–5:1, and the total inoculum size is 1 × 10⁻⁶. 6 ~1×10 8 CFU / mL.

9. The use of a jujube fermentation composition in anti-fatigue and sleep improvement, characterized in that: The jujube fermentation composition described above can play a role in anti-fatigue and improving sleep. The daily intake of the jujube fermentation composition for adults is 200-500g, for children it is 100-200g, and for infants and young children it is 10-50g.

10. The use of the jujube fermentation composition according to claim 9 in anti-fatigue and sleep improvement, characterized in that: The jujube fermentation composition can be further processed into oral liquids, beverages, and jellies to enhance flavor and texture.

Images