Nano-emulsion for improving circadian rhythm disorder sleep disorder and preparation method of nano-emulsion

Abstract

The invention belongs to the technical field of food, and particularly relates to a nano-emulsion for improving circadian rhythm disorder sleep disorder and a preparation method of the nano-emulsion. The semen ziziphi spinosae oil emulsion comprises the following raw materials in parts by weight: 5-10 parts of semen ziziphi spinosae oil, 2-6 parts of rice bran oil, 2-6 parts of wheat germ oil, 0.43-1.15 parts of functional components, 0.78-1.15 parts of an emulsifier and 76-82 parts of water, the emulsifier comprises polyglycerol fatty acid ester, Tween 80 and propylene glycol alginate, and the functional components comprise a black pepper extract, sour cherry powder, sophora flower bud powder and balm. Natural oil and plant extracts are used as core active components, the spina date seed oil has the tranquilizing and allaying excitement effect and has a synergistic effect with unsaturated fatty acid in the rice bran oil and the wheat germ oil, and functional components such as the black pepper extract are matched to regulate neurotransmitter balance; and the problems of difficulty in falling asleep, shallow sleep and the like caused by diurnal rhythm disorder can be accurately improved.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of food technology, and particularly relates to a nanoemulsion for improving sleep disorders of circadian rhythm disorders and a preparation method thereof. BACKGROUND

[0002] Sleep disorders of circadian rhythm disorders are diseases caused by the mismatch between the internal biological clock and the external day-night cycle, resulting in abnormal sleep time and quality. Common symptoms include difficulty falling asleep, early awakening, and daytime sleepiness. Currently, the methods for improving sleep disorders of circadian rhythm disorders mainly include drug intervention and non-drug intervention. Drug intervention mainly relies on sedative and hypnotic chemicals, which can alleviate symptoms in the short term, but long-term use can lead to dependence and drug resistance, and may be accompanied by daytime side effects such as dizziness and sleepiness, posing a high health risk and failing to meet the public's demand for safe sleep improvement.

[0003] Non-drug intervention products have gradually become a research hotspot due to their high safety and long-term consumption advantages. However, existing products still have many technical bottlenecks. On the one hand, most products rely on a single active ingredient (such as melatonin or a single plant extract) to exert their effects, lack synergistic design among multiple components, and are difficult to precisely regulate the complex physiological mechanisms of circadian rhythm disorders (such as neurotransmitter imbalance and lack of unsaturated fatty acids), resulting in limited improvement effects. On the other hand, some active ingredients in the products (such as natural oils and fat-soluble plant extracts) have problems such as bad taste, poor water solubility, and large particle size, which can lead to low bioavailability during the human digestive absorption process, further weakening their actual efficacy.

[0004] Nanoemulsion technology, as a new delivery system, can significantly improve the solubility, stability, and bioavailability of functional ingredients by encapsulating them in nanoscale oil droplets, and has great potential in the field of food functionalization. SUMMARY

[0005] Therefore, the present application provides a nanoemulsion that can improve sleep difficulties and poor sleep quality caused by sleep disorders of circadian rhythm disorders, and has good stability and a long shelf life.

[0006] To solve the above technical problems, the technical solution provided by the present application is as follows:

[0007] The present application provides a nanoemulsion for improving sleep disorders of circadian rhythm disorders, which comprises the following raw materials in parts by weight: 5-10 parts of jujube kernel oil, 2-6 parts of rice bran oil, 2-6 parts of wheat germ oil, 0.43-1.15 parts of functional ingredients, 0.78-1.15 parts of emulsifiers, and 70-90 parts of water.

[0008] Preferably, the functional ingredients include black pepper extract, sour cherry powder, sophora flower bud powder and balm mint.

[0009] Further preferably, the functional ingredients consist of black pepper extract, sour cherry powder, sophora flower bud powder and balm mint.

[0010] More preferably, the weight ratio of black pepper extract, sour cherry powder, sophora flower bud powder and balm mint in the functional ingredients is (0.02-0.05):(0.2-0.5):(0.1-0.4):(0.1-0.5).

[0011] Preferably, the content of piperine in the black pepper extract is not less than 95%.

[0012] Preferably, the extraction ratio of the sour cherry powder is 13:1; that is, 1 kg of sour cherry powder is obtained from 13 kg of fresh sour cherry concentrated and dried.

[0013] Preferably, the content of rosmarinic acid in the balm mint is not less than 6%.

[0014] Preferably, the sophora flower bud powder is prepared by water extraction, concentration and spray drying of the flower buds of Sophora japonica L. in the family Leguminosae, and the concentration ratio is 10:1.

[0015] Preferably, the weight ratio of the sour kernel oil, rice bran oil and wheat germ oil is (5-10):(2-6):(2-6).

[0016] Preferably, the weight ratio of the sour kernel oil to the wheat germ oil is (1.25-3.5):1, and the weight ratio of the sour kernel oil to the rice bran oil is (1-3):1.

[0017] Preferably, the content of jujuboside in the sour kernel oil is not less than 2%.

[0018] Preferably, the emulsifier includes polyglycerol fatty acid ester, Tween 80 and propylene glycol alginate ester.

[0019] Further preferably, the emulsifier includes polyglycerol fatty acid ester 0.6-1 part, Tween 80 0.025-0.05 part and propylene glycol alginate ester 0.05-0.15 part by weight.

[0020] More preferably, the weight ratio of polyglycerol fatty acid ester, Tween 80 and propylene glycol alginate ester is (0.6-1):(0.025-0.05):(0.05-0.15).

[0021] Preferably, the HLB value of the propylene glycol alginate ester is 5-15; further preferably, the HLB value of the propylene glycol alginate ester is 9.

[0022] Preferably, the nanoemulsion for improving circadian rhythm disordered sleep disorder further includes 0.5-2 parts of emulsifier, 0.25-1 part of microcrystalline cellulose, 0.01-0.02 parts of antioxidant and 1-3 parts of fruit juice.

[0023] Preferably, the emulsifier includes glycerin, the antioxidant includes vitamin E, and the juice includes one or more of jujube concentrate, apple concentrate, or white grape concentrate.

[0024] Preferably, the pH of the nanoemulsion that improves circadian rhythm disordered sleep disorder is less than 4.5, and the pH is adjusted by an acid regulator, wherein the acid regulator is citric acid.

[0025] On the other hand, the present invention provides a method for preparing a nanoemulsion that improves circadian rhythm disordered sleep, comprising the following steps:

[0026] (1) Mix jujube seed oil, rice bran oil and wheat germ oil evenly to obtain oil phase A; (2) Mix emulsifier and water, and stir under heating conditions to fully dissolve each component to obtain solution B; (3) Add solution B to oil phase A under stirring conditions to obtain solution C; (4) Emulsify solution C by high-speed shearing, and add the functional component to solution C during the emulsification process to obtain solution D; then perform high-pressure homogenization to obtain the nanoemulsion that improves circadian rhythm disordered sleep disorders.

[0027] Preferably, in step (2), the heating temperature is 70-85℃ and the rotation speed is 60-250 r / min.

[0028] Preferably, in step (4), the rotation speed of high-speed shear emulsification is 6500-8500 r / min, and the emulsification time is 10-20 min.

[0029] Preferably, the pressure of the high-pressure homogenization process in step (4) is 250-350 bar, and the homogenization time is 15-25 min.

[0030] Compared with the prior art, the present invention has the following advantages:

[0031] 1. The nanoemulsion provided by this invention for improving circadian rhythm disordered sleep disorders uses jujube seed oil, rice bran oil, and wheat germ oil as the main oils. Jujube seed oil has a calming and sedative effect. In the prior art, rice bran oil is often used as an oily liquid excipient to dissolve or uniformly mix drugs, while wheat germ oil is often used to improve the stability or antioxidant properties of other oils. However, in this invention, the inventors appropriately increased the proportion of wheat germ oil, so that wheat germ oil, rice bran oil, and jujube seed oil can work synergistically to improve the problems of difficulty falling asleep and sleep quality in zebrafish with circadian rhythm disordered sleep disorders. This may be based on the fact that the unsaturated fatty acids (such as linoleic acid and α-linolenic acid) rich in rice bran oil and wheat germ oil have an important supporting role in the health of the nervous system, thereby achieving the effect of improving sleep quality.

[0032] 2. The nanoemulsion provided by this invention, which improves circadian rhythm disordered sleep, contains black pepper extract, sour cherry powder, sophora japonica powder, and lemon balm. These functional components work synergistically to enhance the effect of improving circadian rhythm disordered sleep. Specifically, the piperine in the black pepper extract acts as a positive allosteric regulator of γ-aminobutyric acid (GABA) benzodiazepine receptors, promoting sleep by regulating GABA secretion. It also counteracts the excitatory effect of caffeine by antagonizing adenosine receptors, thus prolonging sleep time. Melatonin is a key signal regulating the human biological clock and sleep-wake cycle. Sour cherry is one of the few foods that naturally contain melatonin. Exogenous supplementation with substances related to melatonin synthesis can enhance the body's own sleep regulation ability. Sophora japonica powder, through the anti-inflammatory and antioxidant effects of its core components rutin and quercetin, indirectly eliminates some physiological factors that interfere with sleep (such as physical discomfort and inflammation), thereby creating better conditions for sleep. Lemon balm can promote the synthesis of γ-aminobutyric acid (GABA) and partially mimic the effects of GABA by binding to GABA receptors, thereby enhancing inhibitory signals in the brain, reducing neuronal over-excitation, making people feel calm and peaceful, exerting a sedative effect, and making it easier to fall asleep. The four ingredients work together from different angles to improve sleep rhythm disorders.

[0033] 3. The nanoemulsion provided by this invention, which improves sleep disorders caused by circadian rhythm disorders, uses natural oils and plant extracts as its core active components. The calming and sedative effects of jujube seed oil synergize with the unsaturated fatty acids in rice bran oil and wheat germ oil. Combined with functional ingredients such as black pepper extract to regulate neurotransmitter balance, it can precisely improve sleep problems such as difficulty falling asleep and shallow sleep caused by circadian rhythm disorders. Furthermore, the nanoscale particle size significantly enhances bioavailability, providing a safe and highly effective new sleep-improving food for people with circadian rhythm disorder-related insomnia, with promising market application prospects and industrialization value.

[0034] 4. The nanoemulsion provided by this invention improves sleep disorders caused by circadian rhythm imbalance. The emulsifier is a compound of polyglycerol fatty acid ester, Tween 80 and propylene glycol alginate, and the proportions of each component are reasonable. Combined with a co-emulsifier and a specific preparation process, the stability of the emulsion can be significantly improved. Moreover, the average particle size of the nanoemulsion is 200 nm. The nanoscale particle size makes the emulsion more dispersed, which is conducive to the absorption of the active ingredients by the human body and improves bioavailability.

[0035] 5. The nanoemulsion provided by this invention, which improves sleep disorders caused by circadian rhythm disorders, has the addition of microcrystalline cellulose, which can physically stabilize the emulsion through steric hindrance and network structure, providing excellent long-term stability and suspension ability, and preventing emulsion stratification, separation and aggregation.

[0036] 6. The nanoemulsion provided by this invention, which improves sleep disorders caused by circadian rhythm disorder, has added antioxidant vitamin E to prevent oil oxidation and extend the product's shelf life; the addition of fruit juice not only improves the slightly bitter taste and greasiness of jujube seed oil, but also increases the product's nutritional value.

[0037] 7. The nanoemulsion provided by this invention has the function of improving sleep disorders caused by circadian rhythm disorder. The nanoemulsion has a pH of less than 4.5. Under acidic conditions, it can inhibit the growth and reproduction of microorganisms, further extending the shelf life of the product. Moreover, citric acid is used to adjust the pH, and the adjustment effect is stable and will not have an adverse effect on the taste and quality of the product.

[0038] 8. The preparation method of the present invention is simple and easy to operate. Through high-speed shear emulsification and high-pressure homogenization, nanoemulsions with uniform particle size and good stability can be prepared, which are suitable for industrial production. Attached Figure Description

[0039] Figure 1 The image shows the stability of the sample after the nanoemulsion has been stored for 30 days. Detailed Implementation

[0040] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes should also fall within the scope of protection claimed by the present invention.

[0041] When numerical ranges are given in the embodiments, it should be understood that, unless otherwise stated in the invention, both endpoints of each numerical range and any value between the two endpoints may be selected. Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0042] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all chemical reagents used in the embodiments of the present invention are obtained through conventional commercial channels. Products from different manufacturers do not have a significant impact on the effectiveness.

[0043] Experimental instruments and reagents:

[0044] Black pepper extract (piperine ≥95%, Sabinsa Biotechnology Co., Ltd., China); Lemon balm (Moller / rosmarinic acid ≥6%, Shanghai Yipan Biotechnology Co., Ltd.); Sophora japonica powder (Baoji Liupanyun Biotechnology Co., Ltd., 10:1); Sour cherry powder (Shanghai Chengyi Da Health Technology Group Co., Ltd., China); Jujube seed oil (jujube seed saponins ≥2%, China); Rice bran oil (food grade, Heze Zhonghe Jianyuan Biotechnology Co., Ltd., China); Wheat germ oil (food grade, Heze Zhonghe Jianyuan Biotechnology Co., Ltd., China); Glycerin (GLY-995FAG, Wilmar Oils & Fats Technology Co., Ltd., China); Vitamin E (da-tocopherol, Zhejiang Pharmaceutical Co., Ltd., China); Polyglycerol fatty acid ester (99%, Henan Onest, China); Tween 80 (food grade, Guangdong Runhua Chemical Co., Ltd.); Propylene glycol alginate (HLB value 9), Hebei Lequan Biotechnology Co., Ltd., China.

[0045] Example 1

[0046] This embodiment provides a highly stable nanoemulsion that improves circadian rhythm disorder-related sleep disturbances. By weight, it comprises the following ingredients: 5 parts jujube seed oil, 2 parts rice bran oil, 4 parts wheat germ oil, 0.2 parts sophora japonica powder, 0.5 parts lemon balm, 0.4 parts sour cherry powder, 0.05 parts black pepper extract, 0.6 parts polyglycerol fatty acid ester, 0.05 parts Tween 80, 0.15 parts propylene glycol alginate (HLB value of propylene glycol alginate is 9), 1 part glycerin, 0.5 parts microcrystalline cellulose, 82 parts water, 0.010 parts vitamin E, 1 part jujube concentrate, and 2 parts white grape concentrate.

[0047] The preparation method of the above-mentioned nanoemulsion includes the following steps:

[0048] (1) Mix 5 parts of jujube seed oil, 2 parts of rice bran oil, 4 parts of wheat germ oil, 1 part of glycerol, and 0.010 parts of vitamin E evenly to obtain oil phase A;

[0049] (2) Mix 0.6 parts of polyglycerol fatty acid ester, 0.05 parts of Tween 80, 0.15 parts of propylene glycol alginate, 0.5 parts of microcrystalline cellulose, and 82 parts of water. Stir at 80°C for 20 min to fully dissolve all components to obtain solution B.

[0050] (3) Under stirring conditions, solution B is added to oil phase A to obtain solution C;

[0051] (4) Solution C was emulsified by high-speed shearing at a speed of 7000 r / min. During the emulsification process, 0.2 parts of Sophora japonica powder, 0.5 parts of lemon balm, 0.4 parts of sour cherry powder, 0.05 parts of black pepper extract, 1 part of jujube concentrate, and 2 parts of white grape concentrate were added to solution C. The emulsification time was 10 min to obtain crude emulsion D. Anhydrous citric acid was added to adjust the pH of crude emulsion D to 4.0.

[0052] (5) The crude emulsion D was subjected to high-pressure homogenization at a pressure of 350 bar for 15 min to obtain a nanoemulsion that improves sleep disorders caused by circadian rhythm disorders.

[0053] The final pH of the nanoemulsion is 4.0.

[0054] Example 2

[0055] This embodiment provides a highly stable nanoemulsion that improves circadian rhythm disorder-related sleep disturbances. By weight, it comprises the following ingredients: 10 parts jujube seed oil, 4 parts rice bran oil, 3 parts wheat germ oil, 0.1 parts sophora japonica powder, 0.1 parts lemon balm, 0.2 parts sour cherry powder, 0.03 parts black pepper extract, 1 part polyglycerol fatty acid ester, 0.05 parts Tween 80, 0.10 parts propylene glycol alginate (HLB value of propylene glycol alginate is 9), 2 parts glycerin, 0.25 parts microcrystalline cellulose, 76 parts water, 0.02 parts vitamin E, and 3 parts apple concentrate.

[0056] The preparation method of the above-mentioned nanoemulsion includes the following steps:

[0057] (1) Mix 10 parts of jujube seed oil, 4 parts of rice bran oil, 3 parts of wheat germ oil, 2 parts of glycerol and 0.02 parts of vitamin E evenly to obtain oil phase A;

[0058] (2) Mix 1 part of polyglycerol fatty acid ester, 0.05 part of Tween 80, 0.10 part of propylene glycol alginate, 0.25 part of microcrystalline cellulose, and 76 parts of water. Stir at 85°C for 15 min to fully dissolve the components and obtain solution B.

[0059] (3) Under stirring conditions, solution B is added to oil phase A to obtain solution C;

[0060] (4) Solution C was emulsified by high-speed shearing at a speed of 7500 r / min. During the emulsification process, 0.1 parts of Sophora japonica powder, 0.1 parts of lemon balm, 0.2 parts of sour cherry powder, 0.03 parts of black pepper extract, and 3 parts of apple concentrate were added to solution C. The emulsification time was 15 min to obtain crude emulsion D. Anhydrous citric acid was added to adjust the pH of crude emulsion D to 4.0.

[0061] (5) The crude emulsion D was subjected to high-pressure homogenization at a pressure of 250 bar for 25 min to obtain a nanoemulsion that improves sleep disorders caused by circadian rhythm disorder.

[0062] The final pH of the nanoemulsion is 4.0.

[0063] Example 3

[0064] This embodiment provides a highly stable nanoemulsion that improves circadian rhythm disorder-related sleep disturbances. By weight, it comprises the following ingredients: 6 parts jujube seed oil, 6 parts rice bran oil, 2 parts wheat germ oil, 0.3 parts sophora japonica powder, 0.2 parts lemon balm, 0.5 parts sour cherry powder, 0.02 parts black pepper extract, 0.7 parts polyglycerol fatty acid ester, 0.025 parts Tween 80, 0.05 parts propylene glycol alginate (HLB value of propylene glycol alginate is 9), 0.7 parts glycerin, 1 part microcrystalline cellulose, 81 parts water, 0.01 parts vitamin E, and 1 part white grape concentrate.

[0065] The preparation method of the above-mentioned nanoemulsion includes the following steps:

[0066] (1) Mix 6 parts of jujube seed oil, 6 parts of rice bran oil, 2 parts of wheat germ oil, 0.7 parts of glycerol, and 0.01 parts of vitamin E evenly to obtain oil phase A;

[0067] (2) Mix 0.7 parts of polyglycerol fatty acid ester, 0.025 parts of Tween 80, 0.05 parts of propylene glycol alginate, 1 part of microcrystalline cellulose, and 81 parts of water. Stir at 75°C for 25 min to fully dissolve the components and obtain solution B.

[0068] (3) Under stirring conditions, solution B is added to oil phase A to obtain solution C;

[0069] (4) Solution C was emulsified by high-speed shearing at a speed of 6500 r / min. During the emulsification process, 0.3 parts of Sophora japonica powder, 0.2 parts of lemon balm, 0.5 parts of sour cherry powder, 0.02 parts of black pepper extract, and 1 part of white grape concentrate were added to solution C. The emulsification time was 20 min to obtain crude emulsion D. Anhydrous citric acid was added to adjust the pH of crude emulsion D to 4.0.

[0070] (5) The crude emulsion D was subjected to high-pressure homogenization at a pressure of 250 bar for 20 min to obtain a nanoemulsion that improves sleep disorders caused by circadian rhythm disorder.

[0071] The final pH of the nanoemulsion is 4.0.

[0072] Example 4

[0073] This embodiment provides a highly stable nanoemulsion that improves circadian rhythm disorder-related sleep disturbances. By weight, it comprises the following ingredients: 8 parts jujube seed oil, 3 parts rice bran oil, 6 parts wheat germ oil, 0.4 parts sophora japonica powder, 0.3 parts lemon balm, 0.3 parts sour cherry powder, 0.04 parts black pepper extract, 0.9 parts polyglycerol fatty acid ester, 0.05 parts Tween 80, 0.1 parts propylene glycol alginate (HLB value of propylene glycol alginate is 9), 1.5 parts glycerin, 0.7 parts microcrystalline cellulose, 77 parts water, 0.015 parts vitamin E, 1 part jujube concentrate, and 1 part white grape concentrate.

[0074] The preparation method of the above-mentioned nanoemulsion includes the following steps:

[0075] (1) Mix 8 parts of jujube seed oil, 3 parts of rice bran oil, 6 parts of wheat germ oil, 1.5 parts of glycerin and 0.015 parts of vitamin E evenly to obtain oil phase A;

[0076] (2) Mix 0.9 parts of polyglycerol fatty acid ester, 0.05 parts of Tween 80, 0.1 parts of propylene glycol alginate, 0.7 parts of microcrystalline cellulose, and 77 parts of water. Stir at 70°C for 30 minutes to fully dissolve all components and obtain solution B.

[0077] (3) Under stirring conditions, solution B is added to oil phase A to obtain solution C;

[0078] (4) Solution C was subjected to high-speed shear emulsification at a rotation speed of 8500 r / min. During the emulsification process, 0.4 parts of Sophora japonica powder, 0.3 parts of lemon balm, 0.3 parts of sour cherry powder, 0.04 parts of black pepper extract, 1 part of jujube concentrate, and 1 part of white grape concentrate were added to solution B and mixed evenly. The emulsification time was 10 min to obtain crude emulsion D. Anhydrous citric acid was added to adjust the pH of crude emulsion D to 4.0.

[0079] (5) The crude emulsion D was subjected to high-pressure homogenization at a pressure of 300 bar for 15 min to obtain a nanoemulsion that improves sleep disorders caused by circadian rhythm disorders.

[0080] The final pH of the nanoemulsion is 4.0.

[0081] Comparative Example 1

[0082] This comparative example provides a nanoemulsion that improves sleep disorders caused by circadian rhythm disorders. The only difference from Example 2 is that rice bran oil is replaced with jujube seed oil, i.e., 14 parts jujube seed oil, 3 parts wheat germ oil, 0.43 parts functional ingredients (0.1 parts Sophora japonica powder, 0.1 parts lemon balm, 0.2 parts sour cherry powder, 0.03 parts black pepper extract), 1.15 parts emulsifier (including 1 part polyglycerol fatty acid ester, 0.05 parts Tween 80, 0.1 parts propylene glycol alginate (the HLB value of propylene glycol alginate is 9)), 2 parts glycerin, 0.25 parts microcrystalline cellulose, 76 parts water, 0.020 parts vitamin E, and 3 parts apple concentrate.

[0083] The preparation method is the same as in Example 2.

[0084] The final pH of the nanoemulsion is 4.0.

[0085] Comparative Example 2

[0086] This comparative example provides a nanoemulsion that improves sleep disorders caused by circadian rhythm disorders. The only difference from Example 2 is that wheat germ oil is replaced with jujube seed oil, i.e., 13 parts jujube seed oil, 4 parts rice bran oil, 0.43 parts functional ingredients (0.1 parts Sophora japonica powder, 0.1 parts lemon balm, 0.2 parts sour cherry powder, 0.03 parts black pepper extract), 1.15 parts emulsifier (including 1 part polyglycerol fatty acid ester, 0.05 parts Tween 80, 0.1 parts propylene glycol alginate (the HLB value of propylene glycol alginate is 9)), 2 parts glycerin, 0.25 parts microcrystalline cellulose, 76 parts water, 0.020 parts vitamin E, and 3 parts apple concentrate.

[0087] The preparation method is the same as in Example 2.

[0088] The final pH of the nanoemulsion is 4.0.

[0089] Comparative Example 3

[0090] This comparative example provides a nanoemulsion that improves sleep disorders caused by circadian rhythm disorders. The only difference from Example 2 is that the black pepper extract is replaced with lemon balm, namely, 10 parts of jujube seed oil, 4 parts of rice bran oil, 3 parts of wheat germ oil, 0.43 parts of functional ingredients (0.1 parts of sophora japonica powder, 0.13 parts of lemon balm, and 0.2 parts of sour cherry powder), 1.15 parts of emulsifier (including 1 part of polyglycerol fatty acid ester, 0.05 parts of Tween 80, and 0.1 parts of propylene glycol alginate (the HLB value of propylene glycol alginate is 9)), 2 parts of glycerin, 0.25 parts of microcrystalline cellulose, 76 parts of water, 0.020 parts of vitamin E, and 3 parts of apple concentrate.

[0091] The preparation method is the same as in Example 2.

[0092] The final pH of the nanoemulsion is 4.0.

[0093] Comparative Example 4

[0094] This comparative example provides a nanoemulsion that improves sleep disorders caused by circadian rhythm disorders. The only difference from Example 2 is that lemon balm is replaced with black pepper extract, namely, 10 parts of jujube seed oil, 4 parts of rice bran oil, 3 parts of wheat germ oil, 0.43 parts of functional ingredients (0.10 parts of sophora japonica powder, 0.2 parts of sour cherry powder, and 0.13 parts of black pepper extract), 1.15 parts of emulsifier (including 1 part of polyglycerol fatty acid ester, 0.05 parts of Tween 80, and 0.1 parts of propylene glycol alginate (the HLB value of propylene glycol alginate is 9)), 2 parts of glycerin, 0.25 parts of microcrystalline cellulose, 76 parts of water, 0.020 parts of vitamin E, and 3 parts of apple concentrate.

[0095] The preparation method is the same as in Example 2.

[0096] The final pH of the nanoemulsion is 4.0.

[0097] Comparative Example 5

[0098] This comparative example provides a nanoemulsion that improves sleep disorders caused by circadian rhythm disorders. The only difference from Example 2 is that the Sophora japonica powder is replaced with lemon balm, namely, 10 parts of jujube seed oil, 4 parts of rice bran oil, 3 parts of wheat germ oil, 0.43 parts of functional ingredients (0.2 parts of lemon balm, 0.2 parts of sour cherry powder, and 0.03 parts of black pepper extract), 1.15 parts of emulsifier (including 1 part of polyglycerol fatty acid ester, 0.05 parts of Tween 80, and 0.1 parts of propylene glycol alginate (the HLB value of propylene glycol alginate is 9)), 2 parts of glycerin, 0.25 parts of microcrystalline cellulose, 76 parts of water, 0.020 parts of vitamin E, and 3 parts of apple concentrate.

[0099] The preparation method is the same as in Example 2.

[0100] The final pH of the nanoemulsion is 4.0.

[0101] Comparative Example 6

[0102] This comparative example provides a nanoemulsion that improves sleep disorders caused by circadian rhythm disorders. The only difference from Example 2 is that the tart cherry powder is replaced with black pepper extract, namely, 10 parts jujube seed oil, 4 parts rice bran oil, 3 parts wheat germ oil, 0.43 parts functional ingredients (0.1 parts sophora japonica powder, 0.1 parts lemon balm, 0.23 parts black pepper extract), 1.15 parts emulsifier (including 1 part polyglycerol fatty acid ester, 0.05 parts Tween 80, 0.1 parts propylene glycol alginate (the HLB value of propylene glycol alginate is 9)), 2 parts glycerin, 0.25 parts microcrystalline cellulose, 76 parts water, 0.020 parts vitamin E, and 3 parts apple concentrate.

[0103] The preparation method is the same as in Example 2.

[0104] The final pH of the nanoemulsion is 4.0.

[0105] Comparative Example 7

[0106] This comparative example provides a nanoemulsion that improves sleep disorders caused by circadian rhythm disorders. The only difference between this example and Example 2 is that propylene glycol alginate is replaced with sucrose fatty acid ester, namely, 10 parts of jujube seed oil, 4 parts of rice bran oil, 3 parts of wheat germ oil, 0.43 parts of functional ingredients (0.1 parts of sophora japonica powder, 0.1 parts of lemon balm, 0.2 parts of sour cherry powder, and 0.03 parts of black pepper extract), 1.15 parts of emulsifier (including 1 part of polyglycerol fatty acid ester, 0.05 parts of Tween 80, and 0.1 parts of sucrose fatty acid ester (the HLB value of sucrose fatty acid ester is 9)), 2 parts of glycerin, 0.25 parts of microcrystalline cellulose, 76 parts of water, 0.020 parts of vitamin E, and 3 parts of apple concentrate.

[0107] The preparation method is the same as in Example 2.

[0108] The final pH of the nanoemulsion is 4.0.

[0109] Sensory evaluation was performed on the emulsion prepared in Experiment Example 1.

[0110] Fifteen professional evaluators were selected to conduct blind evaluations of the emulsions in Examples 1-4. Sensory evaluations included appearance, odor, texture, flavor, and overall liking. The evaluation criteria are shown in Table 1, and the evaluation results are shown in Table 2.

[0111] Table 1 Sensory Evaluation Criteria

[0112]

[0113] Table 2 Sensory evaluation results

[0114]

[0115] As can be seen from the sensory evaluation in Table 2, Examples 1-4 all received high ratings in terms of appearance, odor, texture, flavor, and overall appeal, with the highest score achieved in appearance. Examples 1-4 all scored over 8 points in odor and flavor, indicating that the emulsion prepared by this invention has no obvious oily or slightly bitter taste. This may be because the addition of fruit juice improved the slight oiliness and bitterness of the emulsion, giving it a fresher, fuller, and more layered sensory presentation.

[0116] Experiment Example 2: Stability Observation

[0117] Instruments: The LUMiSizer stability analyzer was purchased from LUM GmbH, Germany, and the HORIBA SZ-100 V2 nanoparticle analyzer was purchased from Horiba Corporation, Japan.

[0118] 2.1 nanoemulsion average particle size

[0119] Five mL of the nanoemulsion samples obtained in Examples 1-4 and Comparative Example 7 were placed in a constant temperature water bath (70°C) and diluted 500 times with 70°C deionized water, then cooled to room temperature. The diluted samples were added to the sample cell of the nanoparticle analyzer, with the sample volume not less than 2 / 3 full. The sample cell was then inserted into the sample slot and the cover of the nanoparticle analyzer was closed for particle size determination. Each sample was measured three times, and the average value was taken. The results are shown in Table 3.

[0120] Table 3. Results of average particle size determination of nanoemulsions

[0121]

[0122] Particle size is a direct parameter affecting emulsion stability. Larger oil droplets typically have a lower density than water and rise to the top more quickly, forming an oil-rich layer and causing emulsion stratification. Smaller particle sizes result in more vigorous Brownian motion, effectively resisting gravity and significantly delaying or even preventing emulsion separation. Table 3 shows that the average particle size of the nanoemulsions in Examples 1-4 is approximately 200 nm, all exhibiting low particle sizes. Smaller particle sizes lead to more significant Brownian motion, indicating that the emulsions prepared in this invention effectively resist gravity, keeping the droplets suspended and significantly delaying stratification.

[0123] 2.2 Storage stability

[0124] The emulsions obtained in Examples 1-4 and Comparative Example 7 were treated at 90°C for half an hour, and then left at room temperature for 30 days. 1 mL of the nanoemulsion was taken from each sample and placed in a centrifuge tube. The tube was then placed in a stability analyzer and centrifuged at 4000 r / min for 15 min to observe whether the nanoemulsion showed oil-water separation.

[0125] Results analysis:

[0126] After observation, the results are as follows: Figure 1 As shown, the nanoemulsion samples obtained in Examples 1-4 were stable without oil-water separation, while Comparative Example 7 showed slight stratification. This may be because the propylene glycol alginate in the formulation of this invention maintains excellent high viscosity even under acidic conditions. When used synergistically with microcrystalline cellulose, the emulsion exhibits stronger resistance to stresses such as mechanical vibration and temperature fluctuations. While sucrose fatty acid esters possess efficient emulsifying capabilities, they do not have a thickening effect. Therefore, after centrifugation at 4000 r / min for 15 min, Comparative Example 7 showed stratification. This demonstrates that the emulsion prepared by this invention has extremely strong suspension stability, can withstand high-temperature treatment, and is very suitable for emulsion products requiring hot filling or long-term room-temperature storage.

[0127] Experimental Example 3: Efficacy Evaluation

[0128] 3.1 Instruments and Reagents

[0129] Zebrafish behavior analysis system (Zebra Lab 3.22.3.31, Viewpoint, France); 6-well plate (Zhejiang Beilanbo Biotechnology Co., Ltd., China); 96-well plate (Nest Biotech, China).

[0130] 3.2 Experimental grouping and preparation of zebrafish circadian rhythm disorder model

[0131] Several zebrafish embryos aged 0-2 hpf (hours post-fertilization) were selected and, after quality screening, divided into a normal control group, a model group, and a nanoemulsion group (Examples 1-4, Comparative Examples 1-6), and a positive control group (estazolam group). Each group of zebrafish embryos was placed in a six-well plate, with 30 embryos per well, and 5 mL of 1×E3 embryo culture medium was added. The six-well plates were then placed in their respective experimental chambers for a seven-day light experiment. The medium was changed daily during the light-bearing period of the normal control group. The zebrafish in the circadian rhythm disorder group after completing the seven-day dynamic light cycle experiment were the obtained zebrafish circadian rhythm disorder model.

[0132] The illumination time for the experimental chambers of the model group and the nanoemulsion group was set as follows: Day 1: 8:00~20:00; Day 2: 8:00~14:00; Day 3: 2:00~8:00, 20:00~24:00; Day 4: 0:00~8:00, 14:00~24:00; Day 5: 0:00~2:00, 20:00~24:00; Day 6: 0:00~14:00; Day 7: 2:00~20:00. The light source was cool white light (1000 lux), simulating sunlight illumination.

[0133] The illumination time for the normal control group test chamber was set as follows: from day one to day seven, illumination was provided from 8:00 to 20:00 each day, i.e., 12L:12D, with the light source being cool white light (1000 lux) to simulate sunlight.

[0134] After the modeling was completed seven days later, the lighting time for each group was set from 8:00 to 20:00 every day, i.e., 12L:12D.

[0135] After modeling, on day 8, the nanoemulsion group was given samples (dose of 25 μL / mL), while the positive control group was given estazolam (dose of 0.02 mg / mL). After one day of treatment at 28℃, sleep patterns were observed from 20:00 to 21:00 on day 9. Specifically, 10 zebrafish were randomly selected from each experimental group, and the time (T) of movement in the dark environment was quantified using a behavior analyzer to observe sleep patterns. After one day of treatment at 28℃, overall sleep patterns were observed from 20:00 to 8:00 on day 9. Ten zebrafish were randomly selected from each experimental group, and the total distance moved by the zebrafish in the dark environment was quantified using a behavior analyzer to observe overall sleep patterns.

[0136] 3.3 Results Analysis

[0137] Table 4. Effects of nanoemulsions on the duration of movement in zebrafish with disordered diurnal rhythms (n=10)

[0138]

[0139] Note: Compared with the model group, "*" means "p<0.05", "**" means "p<0.01", and "***" means "p<0.001"; compared with Example 2, "#" means "p<0.05", "##" means "p<0.01", and "###" means "p<0.001".

[0140] Within the 1-hour observation period, the movement time of the zebrafish model group was 2831 seconds, which was significantly longer than that of the normal control group (1320 seconds) (p<0.001), indicating that the circadian rhythm disorder model was successfully established. Compared with the model group, the movement time of zebrafish in the positive control group was much shorter than that of the zebrafish model group (p<0.001). This indicates that estazolam has an ameliorative effect on circadian rhythm disorder and sleep disturbance in zebrafish.

[0141] Examples 1-4 showed a significant reduction in the movement duration of zebrafish compared to the model group, indicating that the emulsion prepared in this invention can shorten the sleep onset time for people with circadian rhythm disorders and has an improving effect on sleep disorders caused by circadian rhythm disorders.

[0142] The difference between Comparative Example 1 and Example 2 is that rice bran oil was replaced with jujube seed oil. The difference between Comparative Example 2 and Example 2 is that wheat germ oil was replaced with jujube seed oil. As shown in Table 4, the sleep onset time of Comparative Examples 1-2 was significantly higher than that of Example 2, indicating that the synergistic effect of wheat germ oil and rice bran oil with jujube seed oil in this invention can better shorten the sleep onset time of people with circadian rhythm disorders.

[0143] The difference between Comparative Examples 3-6 and Example 2 is that the functional ingredients were replaced. As shown in Table 4, the sleep onset time of Comparative Examples 3-6 was significantly longer than that of Example 2, indicating that the Sophora japonica powder, lemon balm, sour cherry powder and black pepper extract selected in this invention have a synergistic effect in alleviating the difficulty of falling asleep in people with circadian rhythm disorders.

[0144] Table 5. Effects of nanoemulsions on the movement distance of zebrafish with disordered diurnal rhythms (n=10)

[0145]

[0146] Note: Compared with the model group, "*" means "p<0.05", "**" means "p<0.01", and "***" means "p<0.001"; compared with Example 2, "#" means "p<0.05", "##" means "p<0.01", and "###" means "p<0.001".

[0147] The core characteristic of zebrafish sleep is "motor inhibition." When awake, zebrafish are active and move long distances, while during sleep, movement significantly decreases and moves short distances. Therefore, movement distance directly reflects whether the zebrafish are in a sleep state and the continuity of sleep depth. Quantitative data on the effect of nanoemulsion on the total movement distance of zebrafish with circadian rhythm disorders in a dark environment are shown in Table 5. In the model group, the movement distance of zebrafish was 10330 mm, significantly higher than the 7431 mm in the normal control group (p<0.001), indicating a statistically significant difference, suggesting successful model establishment. In Examples 1-4, the movement distance of zebrafish was significantly lower than that of the model group, indicating that the nanoemulsion provided by this invention can improve sleep quality in circadian rhythm disordered sleep disorders, specifically by reducing nighttime awakenings, increasing sleep duration, and prolonging deep sleep.

[0148] Compared with Example 2, Comparative Examples 1-6 all showed varying degrees of increase in total movement distance, indicating that the oils (jujube seed oil, wheat germ oil, and rice bran oil) and functional ingredients (sophora japonica powder, lemon balm, sour cherry powder, and black pepper extract) provided by the present invention can work synergistically to improve sleep quality in circadian rhythm disordered sleep.

[0149] In summary, the nanoemulsion provided by this invention not only shortens the sleep onset time in zebrafish with circadian rhythm disorders but also prolongs the total nighttime sleep time. This demonstrates that the nanoemulsion prepared by this invention can improve the sleep disturbances and poor sleep quality caused by circadian rhythm disorder-induced insomnia in zebrafish. Since the sleep mechanism of zebrafish, such as circadian rhythms, is similar to that of humans, the nanoemulsion prepared by this invention can improve the symptoms of circadian rhythm disorder-induced sleep disorders in zebrafish, providing a new solution for people with circadian rhythm disorder-induced sleep disorders.

[0150] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A nanoemulsion for improving sleep disorders with circadian rhythm disturbances, characterized in that, By weight parts, including the following raw materials: 5-10 parts of Chinese date kernel oil, 2-6 parts of rice bran oil, 2-6 parts of wheat germ oil, 0.43-1.15 parts of functional ingredients, 0.78-1.15 parts of emulsifiers and 70-90 parts of water; The functional ingredients include black pepper extract, sour cherry powder, sophora powder and balm mint; The emulsifiers include polyglycerol fatty acid ester, Tween 80 and propylene glycol alginate.

2. The nanoemulsion for ameliorating circadian rhythm-related sleep disorders according to claim 1, wherein The weight ratio of polyglycerol fatty acid ester, Tween 80 and propylene glycol alginate in the emulsifiers is (0.6-1):(0.025-0.05):(0.05-0.15).

3. The nanoemulsion for ameliorating circadian rhythm sleep disorder according to claim 1, wherein The weight ratio of black pepper extract, sour cherry powder, sophora powder and balm mint in the functional ingredients is (0.02-0.05):(0.2-0.5):(0.1-0.4):(0.1-0.5).

4. The nanoemulsion for ameliorating circadian rhythm sleep disorder according to claim 1, wherein The weight ratio of Chinese date kernel oil, rice bran oil and wheat germ oil is (5-10):(2-6):(2-6).

5. The nanoemulsion for ameliorating circadian rhythm sleep disorder according to claim 1, wherein The weight ratio of Chinese date kernel oil to wheat germ oil is (1.25-3.5):1; the weight ratio of Chinese date kernel oil to rice bran oil is (1-3):

1.

6. The nanoemulsion for ameliorating circadian rhythm sleep disorder according to claim 1, wherein It also includes 0.5-2 parts of co-emulsifiers, 0.25-1 parts of microcrystalline cellulose, 0.01-0.02 parts of antioxidants and 1-3 parts of fruit juice.

7. The nanoemulsion for ameliorating circadian rhythm sleep disorder according to claim 6, wherein The co-emulsifiers include glycerol; the antioxidants include vitamin E; and the fruit juice includes one or more of red date concentrate, apple concentrate or white grape concentrate.

8. A method of preparing a nanoemulsion having improved sleep disorders of circadian rhythm misalignment according to any one of claims 1 to 7, characterized in that, It includes the following steps: (1) mixing Chinese date kernel oil, rice bran oil and wheat germ oil uniformly to obtain oil phase A; (2) mixing emulsifiers and water, stirring under heating conditions to fully dissolve each ingredient to obtain solution B; (3) under stirring conditions, adding solution B into oil phase A to obtain solution C; (4) high-speed shearing emulsification of solution C, and adding the functional ingredients into solution C during the emulsification process to obtain solution D; Then high-pressure homogenization treatment is carried out to obtain the nanoemulsion for improving circadian rhythm disorder sleep disorders.

9. The preparation method according to claim 8, characterized in that, In step (2), the heating temperature is 70-85℃ and the rotation speed is 60-250 r / min.

10. The preparation method according to claim 8, characterized in that, In step (4), the rotation speed of high-speed shearing emulsification is 6500-8500 r / min and the emulsification time is 10-20 min; the pressure of high-pressure homogenization treatment is 250-350 bar and the homogenization time is 15-25 min.

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