Preparation method and application of nerve-soothing and sleep-aiding composition

Abstract

The invention provides a preparation method and application of a nerve soothing and sleep aiding composition. The compound is prepared from the following raw material medicines in parts by weight: 50 to 60 parts of wild lucid ganoderma extract, 20 to 30 parts of beta-nicotinamide mononucleotide, 2 to 10 parts of coenzyme Q10, 2 to 10 parts of wild cordyceps sinensis and 1 to 10 parts of selenium-enriched yeast. The preparation method comprises the following steps: adding a phosphate buffer solution into wild lucid ganoderma to adjust the pH value, performing ultrasonic hypoxia extraction, centrifugal separation and filtration, taking supernate, passing through a sephadex column, concentrating and drying to obtain wild lucid ganoderma powder. Effective components of the wild ganoderma lucidum extract are increased through sephadex column filtration, and deterioration and oxidation of the effective components in the extraction process are reduced through ultrasonic low-oxygen extraction. According to the application of the nerve-soothing and sleep-aiding composition provided by the invention in the aspects of treating insomnia and relieving fatigue, the preparation method of the composition is simple and easy to operate and implement, and the obtained product has obvious nerve-soothing and sleep-aiding effects.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicines, and in particular to a preparation method and application of a tranquilizing and sleeping-aiding composition. Background Art

[0002] The accelerated pace of social life has led to more and more people experiencing sleep disorders. According to statistics, more than 57% of adults in my country suffer from sleep disorders. Long-term sleep disorders affect normal work and quality of life, cause mental or physical discomfort, and even induce and aggravate cognitive impairment. Currently, tranquilizers and sleep-inducing drugs have the following main disadvantages. First, tranquilizers and hypnotic drugs have the advantage of rapid effect, but long-term use carries the risk of addiction and rebound after drug withdrawal. Secondly, physical hypnosis therapy has also attracted much attention, but it has the disadvantages of a long treatment cycle and unstable treatment effects. At the same time, food ingredients with medicinal and edible properties have been proven to have sedative and tranquilizing effects and have attracted much attention from patients. Traditional Chinese medicine tonic compositions have the function of regulating the body's overall nerves and metabolism, but there is a problem of low utilization rate of effective ingredients. Therefore, in order to solve the above problems, the present invention provides a method for preparing a tranquilizer and sleep-inducing composition through multiple experiments, and its application in tranquilizer and sleep-inducing drugs. Summary of the Invention

[0003] In view of this, the present invention provides a preparation method and application of a tranquilizing and sleep-inducing composition.

[0004] The technical solution of the present invention is achieved as follows:

[0005] A method for preparing a tranquilizing and sleep-inducing composition comprises the following raw materials, in parts by weight: 50-60 parts of wild Ganoderma lucidum extract, 2-10 parts of wild Cordyceps sinensis, 20-30 parts of β-nicotinamide mononucleotide, 2-10 parts of coenzyme Q10, and 1-10 parts of selenium-enriched yeast. The method comprises the following steps: weighing the wild Ganoderma lucidum extract and the wild Cordyceps sinensis, mixing them, sieving them, concentrating them, adding the β-nicotinamide mononucleotide, coenzyme Q10, and selenium-enriched yeast to obtain a mixture, steam-sterilizing the mixture at low temperature, spray-drying it, sieving it, magnetically separating it, and vacuum packaging it to obtain the finished product.

[0006] Furthermore, the sieving is performed through 200-240 mesh, and the concentrating is performed, and the relative density of the concentrated liquid at 60° C. is 1.10-1.15.

[0007] Furthermore, the steam low-temperature sterilization has a preheating temperature of 50-55°C, a preheating time of 20-25 minutes, a sterilization temperature of 80-85°C, and a sterilization time of 50-55 minutes.

[0008] Furthermore, the spray drying has an air inlet temperature of 175-180°C and an air outlet temperature of 90-95°C.

[0009] A method for preparing a tranquilizing and sleep-inducing composition. The preparation of the wild Ganoderma lucidum extract comprises the following steps: taking 50-60 g of wild Ganoderma lucidum, slicing it, adding 110-150 mL of water at 45-55° C., beating the mixture, adding phosphate buffer to adjust the pH to 7.5-8 to obtain wild Ganoderma lucidum slurry, extracting the mixture by ultrasonic hypoxia, centrifuging, filtering, taking the supernatant, passing the supernatant through a dextran gel column, concentrating and drying the supernatant at 60-65° C. to obtain wild Ganoderma lucidum powder, and storing the mixture at low temperature.

[0010] Furthermore, the wild Ganoderma lucidum extract is filtered through an ultrafiltration membrane with a molecular weight cutoff of 2000-3500Da, and the centrifugal separation is performed at a temperature of 15-30°C, a rotation speed of 500-800 rpm / min, and a time of 3-6 minutes.

[0011] Furthermore, the ultrasonic low-oxygen extraction has an extraction temperature of 30-45° C., an extraction time of 25-30 min, a standing time of 10-15 min, a frequency of 20-35 kHz, and an oxygen content of less than 5% during extraction.

[0012] Furthermore, the composition of the glucan gel column equilibrium solution in the preparation of the wild Ganoderma lucidum powder is: 20-23mM Hepes-NaOH, 1.5-2mM Na2-EDTA, 4-6mM MgCl2, pH 7.5-8. Compared with the prior art, the present invention has the following beneficial effects:

[0013] The wild Ganoderma lucidum in the tranquilizing and sleep-inducing composition of the present invention is separated by ultrasonic hypoxia extraction, passed through a dextran gel column, concentrated and dried to obtain wild Ganoderma lucidum powder. The present invention increases the effective ingredients of the wild Ganoderma lucidum extract by filtering through a dextran gel column, and reduces the deterioration and oxidation of the effective ingredients during the extraction process by ultrasonic hypoxia extraction. The tranquilizing and sleep-inducing composition provided by the present invention is used in the treatment of insomnia and the relief of fatigue. The preparation method of the composition is simple and easy to operate, and the obtained product has obvious tranquilizing and sleep-inducing effects. The compound of wild Ganoderma lucidum, wild Cordyceps sinensis and selenium-enriched yeast of the present invention increases the content of 5-HT monoamine neurotransmitters in brain tissue, improves insomnia symptoms, increases the content of inhibitory neurotransmitters GABA, reduces the body's excitement DA value, shortens the time to fall asleep, and increases the time of continuous sleep. DETAILED DESCRIPTION

[0014] In order to better understand the technical content of the present invention, the following specific examples are provided to further illustrate the present invention. Unless otherwise specified, the experimental methods used in the examples of the present invention are all conventional methods.

[0015] Unless otherwise specified, the materials and reagents used in the examples of the present invention are commercially available. The chemical name of Coenzyme Q10 is 2-[(all-E)3,7,11,15,19,23,27,31,35,39-decamethyl-2,6,10,14,18,22,26,30,34,38-tetradecenyl}-5,6-dimethoxy-3-methyl-p-benzoquinone.

[0016] Example 1

[0017] A method for preparing a tranquilizing and sleep-inducing composition, characterized by comprising the following raw materials in parts by weight: 50 parts wild Ganoderma lucidum extract, 5 parts wild Cordyceps sinensis, 20 parts β-nicotinamide mononucleotide, 5 parts coenzyme Q10, and 4 parts selenium-enriched yeast. The method comprises the following steps: weighing the wild Ganoderma lucidum extract and the wild Cordyceps sinensis, mixing them, sieving through a 200-mesh screen, and concentrating the concentrate to a relative density of 1.15 at 60°C; adding β-nicotinamide mononucleotide, the coenzyme, and the selenium-enriched yeast to obtain a mixture; and steam-sterilizing the mixture at low temperature with a preheating temperature of 50°C for 20 minutes, a sterilization temperature of 80°C for 50 minutes, spray drying with an air inlet temperature of 180°C and an air outlet temperature of 95°C, sieving, magnetic separation, and vacuum packaging to obtain the finished product.

[0018] The preparation of wild Ganoderma lucidum extract includes the following steps:

[0019] (1) Take wild Ganoderma lucidum, slice it, add 50℃ water, beat it into pulp, add phosphate buffer to adjust the pH to 7.5, and obtain wild Ganoderma lucidum pulp.

[0020] (2) The wild Ganoderma lucidum slurry was extracted by ultrasonic hypoxia. The wild Ganoderma lucidum slurry was placed in an ultrasonic hypoxia extraction device. The extraction temperature was 35°C, the extraction time was 25 min, and the slurry was allowed to stand for 10 min. The frequency was 30 kHz and the oxygen content during extraction was 3%. After extraction, the slurry was centrifuged at a temperature of 20°C, a rotation speed of 600 rpm / min, and a time of 5 min. The slurry was filtered and passed through a dextran gel column. The composition of the dextran gel column equilibrium solution was: 20 mM Hepes-NaOH, 1.5 mM Na2-EDTA, 4 mM MgCl2, pH 7.5. The slurry was filtered through an ultrafiltration membrane with a molecular weight cutoff of 3000 Da, and the supernatant was taken.

[0021] (3) The supernatant was concentrated and dried at 60°C to obtain wild Ganoderma lucidum powder, which was stored at low temperature.

[0022] Example 2

[0023] A method for preparing a tranquilizing and sleep-inducing composition, characterized by comprising the following raw materials in parts by weight: 60 parts wild Ganoderma lucidum extract, 6 parts wild Cordyceps sinensis, 25 parts β-nicotinamide mononucleotide, 3 parts coenzyme Q10, and 5 parts selenium-enriched yeast. The method comprises the following steps: weighing the wild Ganoderma lucidum extract and the wild Cordyceps sinensis, mixing them, sieving through a 200-mesh screen, and concentrating the concentrate to a relative density of 1.10 at 60°C; adding β-nicotinamide mononucleotide, the coenzyme, and the selenium-enriched yeast to obtain a mixture; and steam-sterilizing the mixture at low temperature with a preheating temperature of 55°C for 25 minutes, a sterilization temperature of 85°C for 55 minutes, spray drying with an air inlet temperature of 180°C and an air outlet temperature of 95°C, sieving, magnetic separation, and vacuum packaging to obtain the finished product.

[0024] The preparation of wild Ganoderma lucidum extract comprises the following steps:

[0025] (1) Take wild Ganoderma lucidum, slice it, add 55℃ water, beat it into pulp, add phosphate buffer to adjust the pH to 8, and obtain wild Ganoderma lucidum pulp.

[0026] (2) The wild Ganoderma lucidum slurry was extracted by ultrasonic hypoxia. The wild Ganoderma lucidum slurry was placed in an ultrasonic hypoxia extraction device, the extraction temperature was 45°C, the extraction time was 30 min, the extraction was allowed to stand for 15 min, the frequency was 35 kHz, the oxygen content during extraction was 4%, and the extraction was centrifuged at a temperature of 30°C, a rotation speed of 800 rpm / min, and a time of 6 min. The extraction was filtered and passed through a dextran gel column. The composition of the dextran gel column equilibrium solution was: 23 mM Hepes-NaOH, 2 mM Na2-EDTA, 6 mM MgCl2, pH 7.5. The filter membrane was filtered through an ultrafiltration membrane with a molecular weight cutoff of 3500 Da, and the supernatant was taken.

[0027] (3) The supernatant was concentrated and dried at 60°C to obtain wild Ganoderma lucidum powder, which was stored at low temperature.

[0028] Example 3

[0029] A method for preparing a tranquilizing and sleep-inducing composition, characterized by comprising the following raw materials in parts by weight: 55 parts of wild Ganoderma lucidum extract, 3 parts of wild Cordyceps sinensis, 250 parts of β-nicotinamide mononucleotide, 6 parts of coenzyme Q10, and 10 parts of selenium-enriched yeast. The method comprises the following steps: weighing the wild Ganoderma lucidum extract and the wild Cordyceps sinensis, mixing them, sieving through a 200-mesh screen, and concentrating the concentrate to a relative density of 1.10 at 60°C; adding β-nicotinamide mononucleotide, coenzyme Q10, and selenium-enriched yeast to obtain a mixture; and steam-sterilizing the mixture at low temperature with a preheating temperature of 52°C for 23 minutes, a sterilization temperature of 85°C for 50 minutes, spray drying with an air inlet temperature of 180°C and an air outlet temperature of 95°C, sieving, magnetic separation, and vacuum packaging to obtain the finished product.

[0030] The preparation of wild Ganoderma lucidum extract includes the following steps:

[0031] (1) Take wild Ganoderma lucidum, slice it, add 45℃ water, beat it into pulp, add phosphate buffer to adjust the pH to 7.5, and obtain wild Ganoderma lucidum pulp.

[0032] (2) The wild Ganoderma lucidum slurry was extracted by ultrasonic hypoxia. The wild Ganoderma lucidum slurry was placed in an ultrasonic hypoxia extraction device. The extraction temperature was 30°C, the extraction time was 25 min, and the slurry was allowed to stand for 10 min. The frequency was 20 kHz and the oxygen content during extraction was 4%. After extraction, the slurry was centrifuged at a temperature of 15°C, a rotation speed of 500 rpm / min, and a time of 3 min. The slurry was filtered and passed through a dextran gel column. The composition of the dextran gel column equilibrium solution was: 20 mM Hepes-NaOH, 1.5 mM Na2-EDTA, 4 mM MgCl2, pH 7.5. The slurry was filtered through an ultrafiltration membrane with a molecular weight cutoff of 2000 Da, and the supernatant was collected.

[0033] (3) The supernatant was concentrated and dried at 60°C to obtain wild Ganoderma lucidum powder, which was stored at low temperature.

[0034] Comparative Example 1

[0035] The difference from Example 1 is that the filtration step of the wild Ganoderma lucidum extraction does not use an ultrafiltration membrane, but uses filter paper for filtration, and the dosage of the composition and the preparation method are the same as those in Example 1.

[0036] The preparation of wild Ganoderma lucidum extract includes the following steps:

[0037] (1) Take wild Ganoderma lucidum, slice it, add 50℃ water, beat it into pulp, add phosphate buffer to adjust the pH to 7.5, and obtain wild Ganoderma lucidum pulp.

[0038] (2) The wild Ganoderma lucidum slurry was extracted by ultrasonic hypoxia. The wild Ganoderma lucidum slurry was placed in an ultrasonic hypoxia extraction device, the extraction temperature was 35°C, the extraction time was 25 min, the extraction was allowed to stand for 10 min, the frequency was 30 kHz, the oxygen content during extraction was 3%, and the extraction was followed by centrifugation at a temperature of 20°C, a rotation speed of 600 rpm / min, a time of 5 min, and then filtered with filter paper.

[0039] (3) The supernatant was concentrated and dried at 60°C to obtain wild Ganoderma lucidum powder, which was stored at low temperature.

[0040] Comparative Example 2

[0041] The difference from Example 1 is that in the step of extracting wild Ganoderma lucidum, the content of the glucan gel column components is modified to 10mM Hepes-NaOH, 0.5mM Na2-EDTA, 3mM MgCl2, pH 6.5, and the dosage and preparation method of the composition are consistent with those in Example 1.

[0042] The preparation of wild Ganoderma lucidum extract comprises the following steps:

[0043] (1) Take wild Ganoderma lucidum, slice it, add 55℃ water, beat it into pulp, add phosphate buffer to adjust the pH to 8, and obtain wild Ganoderma lucidum pulp.

[0044] (2) The wild Ganoderma lucidum slurry was extracted by ultrasonic hypoxia. The wild Ganoderma lucidum slurry was placed in an ultrasonic hypoxia extraction device. The extraction temperature was 45°C, the extraction time was 30 min, and the slurry was allowed to stand for 15 min at a frequency of 35 kHz. The oxygen content during extraction was 4%. After extraction, the slurry was centrifuged at a temperature of 30°C, a rotation speed of 800 rpm / min, and a time of 6 min. The slurry was filtered and passed through a dextran gel column. The composition of the dextran gel column equilibrium solution was: 10 mM Hepes-NaOH, 0.5 mM Na2-EDTA, 3 mM MgCl2, pH 6.5. The slurry was filtered through an ultrafiltration membrane with a molecular weight cutoff of 3500 Da, and the supernatant was collected.

[0045] (3) The supernatant was concentrated and dried at 60°C to obtain wild Ganoderma lucidum powder, which was stored at low temperature.

[0046] Comparative Example 3

[0047] The difference from Example 1 is that selenium-enriched yeast is removed from the raw materials, and other raw material proportions and preparation methods are consistent with Example 1.

[0048] Comparative Example 4

[0049] The difference from Example 1 is that in the step of extracting wild Ganoderma lucidum, ultrasonic low-oxygen extraction is changed to ultrasonic conventional oxygen extraction, and the dosage of the composition and the preparation method are consistent with those in Example 1.

[0050] The preparation of wild Ganoderma lucidum extract comprises the following steps:

[0051] (1) Take wild Ganoderma lucidum, slice it, add 50℃ water, beat it into pulp, add phosphate buffer to adjust the pH to 7.5, and obtain wild Ganoderma lucidum pulp.

[0052] (2) The wild Ganoderma lucidum slurry was extracted by ultrasonic hypoxia. The wild Ganoderma lucidum slurry was ultrasonically extracted at an extraction temperature of 35°C, an extraction time of 25 min, and a standing time of 10 min at a frequency of 30 kHz. The oxygen content during extraction was the laboratory oxygen content. After extraction, the slurry was centrifuged at a centrifugal temperature of 20°C, a rotation speed of 600 rpm / min, and a time of 5 min. The slurry was filtered and passed through a dextran gel column. The composition of the dextran gel column equilibrium solution was: 20 mM Hepes-NaOH, 1.5 mM Na2-EDTA, 4 mM MgCl2, pH 7.5. The slurry was filtered through an ultrafiltration membrane with a molecular weight cutoff of 3000 Da, and the supernatant was collected.

[0053] (3) The supernatant was concentrated and dried at 60°C to obtain wild Ganoderma lucidum powder, which was stored at low temperature.

[0054] Positive control:

[0055] Tranquilizing sleep-aiding oral liquid, specification: 15mL / tube, manufacturer: Shenzhen Taitai Pharmaceutical Co., Ltd.

[0056] Blank control:

[0057] An equal amount of purified water was taken as a blank control.

[0058] Table 1

[0059]

[0060]

[0061] Experimental study: 70 healthy mice were randomly selected and divided into 7 groups of 10 mice each. The mice weighed 20-25g. The drug was administered via gavage at a dose of 0.2mL / 10g between 8pm and 9pm. The administration cycle lasted 4 courses, each lasting 7 days.

[0062] (1) Sleep test:

[0063] After 28 days of continuous dosing, the mice's sleep was observed. The duration of loss of righting reflex (LOR) was recorded. LOR refers to the inability of mice to right themselves for more than 60 seconds when in dorsal recumbency. The time from loss of righting reflex to recovery was considered sleep duration. The number of mice that fell asleep and the duration of sleep were observed and recorded for each group (see Table 2).

[0064] Table 2

[0065]

[0066] After 28 days of the experiment, the sleeping status of the mice after 5 minutes, 10 minutes and 15 minutes was statistically analyzed, and it was found that the tranquilizing and sleeping-aiding composition prepared by the raw materials, proportions and preparation method of the present invention was easier to fall asleep after taking, and the continuous sleeping time was increased.

[0067] (2) Brain tissue test:

[0068] This study measured the levels of 5-HT, GABA, and DA in mouse brain tissue (see Table 3). 5-HT is a monoamine neurotransmitter involved in sleep regulation; increased 5-HT levels can improve insomnia symptoms. GABA is the main inhibitory neurotransmitter in brain tissue, distributed in the cerebral cortex, hypothalamus, brainstem, hippocampus, and basal ganglia. During sleep, increased GABA release from the cortex promotes sleep, and activation of GABA receptors promotes sleep. DA plays an important role in motor control, reward, emotional regulation, and addictive behaviors in the human body. Excessive DA levels can cause a sense of excitement, leading to insomnia.

[0069] Table 3

[0070]

[0071] Results from a mouse brain tissue test showed that 28 days of continuous administration of the tranquilizing and sleep-inducing composition of the present invention increased 5-HT and GABA levels in the mouse brain tissue, while decreasing DA levels. Compared to Comparative Examples 1-4, Example 1 showed an increase of 3.2 pg / mg in 5-HT, 186.7 pg / mg in GABA, and a decrease of 0.55 pg / mg in DA.

[0072] Conclusion: The tranquilizing and sleep-inducing composition of the present invention has a significant effect on regulating sleep. After continuous use for 28 days, the tranquilizing and sleep-inducing effects of Examples 1-3 are higher than those of the comparative example, the positive control, and the blank control. The preparation process of the wild Ganoderma lucidum extract of the present invention has a better effect. Comparative Example 1 changes the filtration step of the wild Ganoderma lucidum extract and does not use an ultrafiltration membrane. After filtering with filter paper, the increase in 5-HT and GABA and the decrease in DA in the mouse brain tissue are lower than those in Example 1. Comparative Example 2 changes the component content of the dextran gel column when extracting the wild Ganoderma lucidum extract. The mice's sleep time increases, and the 5-HT and GABA content values ​​in the brain tissue are lower. After removing the selenium-rich yeast from the raw materials of Comparative Example 3, the mice's sleep time increases, the 5-HT and GABA content values ​​in the brain tissue are lower, and the DA value is higher. This shows that the tranquilizing and sleep-inducing composition obtained by the raw materials, proportions, and preparation methods of the present invention has a better compound effect and has a significant therapeutic effect in tranquilizing and sleep-inducing.

[0073] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for preparing a tranquilizing and sleep-inducing composition, characterized in that: The invention comprises the following raw materials in parts by weight: 50-60 parts of wild ganoderma lucidum extract, 2-10 parts of wild cordyceps sinensis, 20-30 parts of beta-nicotinamide mononucleotide, 2-10 parts of coenzyme Q10 and 1-10 parts of selenium-enriched yeast, and comprises the following production steps: weighing the wild ganoderma lucidum extract and the wild cordyceps sinensis, mixing, sieving, concentrating, adding beta-nicotinamide mononucleotide, coenzyme and selenium-enriched yeast to obtain a mixture, steam-sterilizing the mixture at low temperature, spray drying, sieving, magnetic separation and vacuum packaging to obtain a finished product.

2. The method for preparing a tranquilizing and sleeping-aiding composition according to claim 1, characterized in that: The sieving is performed through 200-240 meshes, and the concentrating is performed so that the relative density of the concentrated liquid at 60° C. is 1.10-1.

15.

3. The method for preparing a tranquilizing and sleeping-aiding composition according to claim 1, characterized in that: The steam low-temperature sterilization has a preheating temperature of 50-55°C, a preheating time of 20-25 minutes, a sterilization temperature of 80-85°C, and a sterilization time of 50-55 minutes.

4. The method for preparing a tranquilizing and sleeping-aiding composition according to claim 1, characterized in that: The spray drying has an air inlet temperature of 175-180°C and an air outlet temperature of 90-95°C.

5. The method for preparing a tranquilizing and sleeping-aiding composition according to claim 1, characterized in that: The preparation of the wild ganoderma lucidum extract comprises the following steps: taking 50-60 g of wild ganoderma lucidum, slicing, adding 110-150 mL of water at 45-55° C., beating, adding phosphate buffer to adjust the pH to 7.5-8 to obtain wild ganoderma lucidum slurry, extracting by ultrasonic hypoxia, centrifuging, filtering, taking supernatant, passing the supernatant through a dextran gel column, concentrating and drying at 60-65° C. to obtain wild ganoderma lucidum powder, and storing at low temperature.

6. The method for preparing a tranquilizing and sleeping-aiding composition according to claim 5, characterized in that: The wild ganoderma lucidum extract is filtered through an ultrafiltration membrane with a molecular weight cutoff of 2000-3500Da. The centrifugal separation is performed at a centrifugal temperature of 15-30°C, a rotation speed of 500-800rpm / min, and a time of 3-6min.

7. The method for preparing a tranquilizing and sleeping-aiding composition according to claim 5, characterized in that: The ultrasonic low-oxygen extraction has an extraction temperature of 30-45° C., an extraction time of 25-30 min, a standing time of 10-15 min, a frequency of 20-35 kHz, and an oxygen content of less than 5% during extraction.

8. The method for preparing a tranquilizing and sleep-aiding composition according to claim 5, characterized in that: The components of the glucan gel column equilibrium solution in the preparation of the wild ganoderma powder are: 20-23mM Hepes-NaOH, 1.5-2mM Na2-EDTA, 4-6mM MgCl2, pH7.5-8.

9. Use of the composition prepared by the preparation method according to any one of claims 1 to 8 in preparing a drug for conditioning and treating insomnia.

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