Preparation method and application of a composition for tranquilizing the mind and helping sleep

Abstract

The present invention provides a preparation method and application of a composition for calming the nerves and promoting sleep. The components of the complex include the following raw materials in parts by weight: 50-60 parts of wild ganoderma extract, 20-30 parts of β-nicotinamide mononucleotide, 2-10 parts of coenzyme Q10, 2-10 parts of wild cordyceps, and 1-10 parts of selenium-enriched yeast. The pH value of wild ganoderma is adjusted with phosphate buffer solution, and ultrasonic low-oxygen extraction, centrifugal separation, filtration are carried out. The supernatant is taken, passed through a Sephadex column, concentrated and dried to obtain wild ganoderma powder. The present invention increases the active ingredients of wild ganoderma extract through filtration with a Sephadex column, and reduces the deterioration and oxidation of active ingredients during the extraction process through ultrasonic low-oxygen extraction. The application of the composition for calming the nerves and promoting sleep provided by the present invention in the treatment of insomnia and alleviating easy fatigue has the advantages that the preparation method of the composition is simple and easy to operate and implement, and the obtained product has an obvious effect of calming the nerves and promoting sleep.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine, and particularly relates to a preparation method and application of a composition for soothing the nerves and promoting sleep. Background Art

[0002] The accelerating pace of social life has led to sleep disorders in an increasing number of people. According to statistics, more than 57% of adults in China have sleep disorders. Long-term sleep disorders affect normal work and quality of life, cause mental or physical discomfort, and even induce or exacerbate cognitive impairment diseases. At present, the main disadvantages of sedative and hypnotic drugs for soothing the nerves and promoting sleep are as follows. First, sedative and hypnotic drugs have the advantage of quick effect, but long-term use may have risks such as addiction and withdrawal rebound. Second, physical hypnotherapy has also received much attention. However, it has the disadvantages of a long treatment cycle and unstable treatment effect. At the same time, foods homologous to medicine and food have been proven to have the effect of calming the nerves and soothing the mind, and have attracted much attention from patients. The composition of traditional Chinese medicine tonics has the function of regulating the nerves and metabolism of the whole body of the human body. However, there is a problem of low utilization rate of active ingredients. Therefore, in order to solve the above problems, through multiple experiments, the present invention provides a preparation method of a composition for soothing the nerves and promoting sleep and its application in drugs for soothing the nerves and promoting sleep. Summary of the Invention

[0003] In view of this, the present invention provides a preparation method and application of a composition for soothing the nerves and promoting sleep.

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

[0005] A preparation method of a composition for soothing the nerves and promoting sleep, comprising 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 preparation method includes the following steps: weighing the wild ganoderma lucidum extract and wild cordyceps sinensis, mixing, sieving, concentrating, adding β-nicotinamide mononucleotide, coenzyme and selenium-enriched yeast to obtain a mixture, subjecting the mixture to steam low-temperature sterilization, spray drying, sieving, magnetic separation, and vacuum packaging to obtain a finished product.

[0006] Further, for the sieving, it is sieved through 200 - 240 meshes, and for the concentration, the relative density of the concentrated solution is 1.10 - 1.15 at 60°C.

[0007] Further, for the steam low-temperature sterilization, the preheating temperature is 50 - 55°C, the preheating time is 20 - 25 min, the sterilization temperature is 80 - 85°C, and the sterilization time is 50 - 55 min.

[0008] Further, for the spray drying, the inlet air temperature for drying is 175 - 180°C, and the outlet air temperature is 90 - 95°C.

[0009] A preparation method of a composition for soothing the nerves and promoting sleep. The preparation of the wild ganoderma extract comprises the following steps: Take 50 - 60 g of wild ganoderma, slice it, add 110 - 150 mL of water at 45 - 55 °C, make a pulp, add phosphate buffer to adjust the pH to 7.5 - 8 to obtain a wild ganoderma slurry, perform ultrasonic low-oxygen extraction, centrifuge and separate, filter, take the supernatant, pass the supernatant through a Sephadex column, concentrate and dry at 60 - 65 °C to obtain wild ganoderma powder, and store it at low temperature.

[0010] Furthermore, the filtrate of the wild ganoderma extract is filtered through an ultrafiltration membrane with a molecular weight cut-off of 2000 - 3500 Da. For the centrifugal separation, the centrifugation temperature is 15 - 30 °C, the rotation speed is 500 - 800 rpm / min, and the time is 3 - 6 min.

[0011] Furthermore, for the ultrasonic low-oxygen extraction, the extraction temperature is 30 - 45 °C, the extraction time is 25 - 30 min, let it stand for 10 - 15 min, the frequency is 20 - 35 kHz, and the oxygen content during extraction is less than 5%.

[0012] Furthermore, the components of the equilibration solution of the Sephadex column in the preparation of the wild ganoderma powder are: 20 - 23 mM Hepes-NaOH, 1.5 - 2 mM Na 2 -EDTA, 4 - 6 mM MgCl 2 , pH 7.5 - 8. Compared with the prior art, the beneficial effects of the present invention are:

[0013] In the composition for soothing the nerves and promoting sleep of the present invention, the wild ganoderma is extracted and separated by ultrasonic low-oxygen extraction, passed through a Sephadex column, concentrated and dried to obtain wild ganoderma powder. The present invention increases the effective components of the wild ganoderma extract by filtration through a Sephadex column, and reduces the deterioration and oxidation of the effective components during the extraction process by ultrasonic low-oxygen extraction. The application of the composition for soothing the nerves and promoting sleep provided by the present invention in the treatment of insomnia and alleviating easy fatigue has a simple and easy-to-operate preparation method, and the obtained product has an obvious effect of soothing the nerves and promoting sleep. The wild ganoderma, wild cordyceps sinensis and selenium-enriched yeast in the present invention are compounded, which increases the content of 5-HT monoamine neurotransmitters in the brain tissue, improves the insomnia symptoms, increases the content of inhibitory neurotransmitter GABA, reduces the DA value of the body's excitability, shortens the sleep onset time, and increases the duration of continuous sleep. Specific Embodiments

[0014] To better understand the technical content of the present invention, specific examples are provided below to further illustrate the present invention. The experimental methods used in the examples of the present invention are all conventional methods unless otherwise specified.

[0015] Materials, reagents, etc. used in the embodiments of the present invention can be obtained from commercial channels without special instructions. 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-tetracontadecenyl}-5,6-dimethoxy-3-methyl-p-benzoquinone.

[0016] Example 1

[0017] A preparation method of a sedative and sleep-aid composition, characterized by comprising the following raw materials in parts by weight: 50 parts of wild ganoderma lucidum extract, 5 parts of wild cordyceps sinensis, 20 parts of β-nicotinamide mononucleotide, 5 parts of coenzyme Q10, and 4 parts of selenium-enriched yeast. The preparation method includes the following steps: Weigh the wild ganoderma lucidum extract and wild cordyceps sinensis, mix them, sieve through 200 meshes, concentrate, and the relative density of the concentrated solution is 1.15 at 60°C. Add β-nicotinamide mononucleotide, coenzyme, and selenium-enriched yeast to obtain a mixture. Steam-sterilize the mixture at a low temperature, with a preheating temperature of 50°C, a preheating time of 20 min, a sterilization temperature of 80°C, and a sterilization time of 50 min. Spray-dry, with an inlet air temperature of 180°C and an outlet air temperature of 95°C for drying, sieve, perform magnetic separation, and vacuum-pack to obtain the finished product.

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

[0019] (1) Take wild ganoderma lucidum, slice it, add water at 50°C, make a slurry, and adjust the pH to 7.5 with a phosphate buffer solution to obtain a wild ganoderma lucidum slurry.

[0020] (2) The wild ganoderma lucidum slurry is extracted by ultrasonic low-oxygen extraction. Place the wild ganoderma lucidum slurry in an ultrasonic low-oxygen extraction device, with an extraction temperature of 35°C, an extraction time of 25 min, let it stand for 10 min, a frequency of 30 kHz, an oxygen content of 3% during extraction, perform centrifugal separation after extraction, with a centrifugal temperature of 20°C, a rotation speed of 600 rpm / min, and a time of 5 min, filter, and pass through a Sephadex gel column. The composition of the Sephadex gel column equilibration solution is: 20 mM Hepes-NaOH, 1.5 mM Na 2 -EDTA, 4 mM MgCl 2 , pH 7.5, filter through an ultrafiltration membrane with a molecular weight cut-off of 3000 Da, and take the supernatant;

[0021] (3) Concentrate and dry the supernatant at 60°C to obtain wild ganoderma lucidum powder, and store it at a low temperature.

[0022] Example 2

[0023] A preparation method of a soothing sleep aid composition, characterized by comprising the following raw materials in parts by weight: 60 parts of wild ganoderma lucidum extract, 6 parts of wild cordyceps sinensis, 25 parts of β-nicotinamide mononucleotide, 3 parts of coenzyme Q10, and 5 parts of selenium-enriched yeast. The preparation method includes the following steps: Weigh the wild ganoderma lucidum extract and wild cordyceps sinensis, mix them, sieve through 200 meshes, concentrate, and the concentrated liquid has a relative density of 1.10 at 60 °C. Add β-nicotinamide mononucleotide, coenzyme, and selenium-enriched yeast to obtain a mixture. Sterilize the mixture by steam at a low temperature, with a preheating temperature of 55 °C, a preheating time of 25 min, a sterilization temperature of 85 °C, and a sterilization time of 55 min. Spray dry, with an inlet air temperature of 180 °C and an outlet air temperature of 95 °C for drying, sieve, magnetic separation, and vacuum packaging to obtain the finished product.

[0024] The preparation of wild ganoderma lucidum extract includes the following steps:

[0025] (1) Take wild ganoderma lucidum, slice it, add water at 55 °C, make a slurry, and adjust the pH to 8 with phosphate buffer to obtain a wild ganoderma lucidum slurry.

[0026] (2) The wild ganoderma lucidum slurry is extracted by ultrasonic low oxygen. Place the wild ganoderma lucidum slurry in an ultrasonic low oxygen extraction device, with an extraction temperature of 45 °C, an extraction time of 30 min, stand for 15 min, a frequency of 35 kHz, an oxygen content of 4% during extraction, and after extraction, centrifuge at a temperature of 30 °C, a rotation speed of 800 rpm / min, and a time of 6 min, filter, and pass through a Sephadex gel column. The composition of the Sephadex gel column equilibration solution is: 23 mM Hepes-NaOH, 2 mM Na 2 -EDTA, 6 mM MgCl 2 , pH 7.5, filter through an ultrafiltration membrane with a molecular weight cut-off of 3500 Da, and take the supernatant;

[0027] (3) Concentrate and dry the supernatant at 60 °C to obtain wild ganoderma lucidum powder, and store it at low temperature.

[0028] Example 3

[0029] A preparation method of a soothing sleep aid 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 preparation method includes the following steps: Weigh the wild ganoderma lucidum extract and wild cordyceps sinensis, mix them, sieve through 200 meshes, concentrate, and the concentrated liquid has a relative density of 1.10 at 60 °C. Add β-nicotinamide mononucleotide, coenzyme, and selenium-enriched yeast to obtain a mixture. Sterilize the mixture by steam at a low temperature, with a preheating temperature of 52 °C, a preheating time of 23 min, a sterilization temperature of 85 °C, and a sterilization time of 50 min. Spray dry, with an inlet air temperature of 180 °C and an outlet air temperature of 95 °C for drying, sieve, magnetic separation, and vacuum packaging to obtain the finished product.

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

[0031] (1) Take wild Ganoderma lucidum, slice it, add water at 45°C, make a pulp, and adjust the pH to 7.5 with phosphate buffer to obtain wild Ganoderma lucidum pulp.

[0032] (2) The wild Ganoderma lucidum pulp is extracted by ultrasonic low-oxygen extraction. Place the wild Ganoderma lucidum pulp in an ultrasonic low-oxygen extraction device. The extraction temperature is 30°C, the extraction time is 25 min, let it stand for 10 min, the frequency is 20 kHz, the oxygen content during extraction is 4%, after extraction, centrifuge and separate, the centrifuge temperature is 15°C, the rotation speed is 500 rpm / min, the time is 3 min, filter, pass through a Sephadex column, and the composition of the Sephadex column equilibration solution is: 20 mM Hepes-NaOH, 1.5 mM Na 2 -EDTA, 4 mM MgCl 2 , pH 7.5, filter through an ultrafiltration membrane with a molecular weight cut-off of 2000 Da, and take the supernatant;

[0033] (3) Concentrate and dry the supernatant at 60°C to obtain wild Ganoderma lucidum powder, and store it at low temperature.

[0034] Comparative Example 1

[0035] The difference from Example 1 is that: in the filtration step of wild Ganoderma lucidum extraction, filter paper is used instead of an ultrafiltration membrane, and the dosage and preparation method of the composition are the same as those in Example 1.

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

[0037] (1) Take wild Ganoderma lucidum, slice it, add water at 50°C, make a pulp, and adjust the pH to 7.5 with phosphate buffer to obtain wild Ganoderma lucidum pulp.

[0038] (2) The wild Ganoderma lucidum pulp is extracted by ultrasonic low-oxygen extraction. Place the wild Ganoderma lucidum pulp in an ultrasonic low-oxygen extraction device. The extraction temperature is 35°C, the extraction time is 25 min, let it stand for 10 min, the frequency is 30 kHz, the oxygen content during extraction is 3%, after extraction, centrifuge and separate, the centrifuge temperature is 20°C, the rotation speed is 600 rpm / min, the time is 5 min, and filter with filter paper,

[0039] (3) Take the supernatant and concentrate and dry it at 60°C to obtain wild Ganoderma lucidum powder, and store it at low temperature.

[0040] Comparative Example 2

[0041] The difference from Example 1 is that: in the steps of wild Ganoderma lucidum extraction, the content of the Sephadex column components is modified to 10 mM Hepes-NaOH, 0.5 mM Na 2-EDTA, 3 mM MgCl 2 , pH 6.5, the dosage and preparation method of the composition are the same as those in Example 1.

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

[0043] (1) Take wild Ganoderma lucidum, slice it, add water at 55 °C, make a pulp, and adjust the pH to 8 with a phosphate buffer to obtain a wild Ganoderma lucidum slurry.

[0044] (2) The wild Ganoderma lucidum slurry is extracted by ultrasonic low-oxygen extraction. Place the wild Ganoderma lucidum slurry in an ultrasonic low-oxygen extraction device. The extraction temperature is 45 °C, the extraction time is 30 min, let it stand for 15 min, the frequency is 35 kHz, the oxygen content during extraction is 4%, after extraction, centrifuge and separate, the centrifuge temperature is 30 °C, the rotation speed is 800 rpm / min, the time is 6 min, filter, and pass through a Sephadex column. The composition of the Sephadex column equilibration solution is: 10 mM Hepes-NaOH, 0.5 mM Na 2 -EDTA, 3 mM MgCl 2 , pH 6.5, filter through an ultrafiltration membrane with a molecular weight cut-off of 3500 Da, and take the supernatant;

[0045] (3) Concentrate and dry the supernatant at 60 °C to obtain wild Ganoderma lucidum powder and store it 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 the other raw material ratios and production methods are the same as those in Example 1.

[0048] Comparative Example 4

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

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

[0051] (1) Take wild Ganoderma lucidum, slice it, add water at 50 °C, make a pulp, and adjust the pH to 7.5 with a phosphate buffer to obtain a wild Ganoderma lucidum slurry.

[0052] (2) The wild ganoderma lucidum slurry is extracted by ultrasonic low oxygen. The wild ganoderma lucidum slurry is ultrasonically extracted at a temperature of 35 °C for 25 min, allowed to stand for 10 min, with a frequency of 30 kHz. The oxygen content during extraction is the oxygen content in the laboratory. After extraction, it is centrifuged at a temperature of 20 °C, a rotation speed of 600 rpm / min for 5 min, filtered, and passed through a Sephadex column. The composition of the Sephadex column equilibration solution is: 20 mM Hepes-NaOH, 1.5 mM Na 2 -EDTA, 4 mM MgCl 2 , pH 7.5. It is filtered through an ultrafiltration membrane with a molecular weight cut-off of 3000 Da, and the supernatant is taken;

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

[0054] Positive control:

[0055] Jingxin Zhumian Oral Liquid, specification: 15 mL / branch, manufacturer: Shenzhen Taotai Pharmaceutical Co., Ltd.

[0056] Blank control:

[0057] An equal amount of pure water is taken as the blank control.

[0058] Table 1

[0059]

[0060]

[0061] Experiment: A mouse experiment is conducted. 70 healthy mice are randomly selected and divided into 7 groups, with 10 mice in each group. The body weight of the mice is 20 - 25 g, and the gavage dosage is 0.2 mL / 10 g. The medicine is administered at 20 - 21 o'clock in the evening. The administration cycle is 4 courses, and each course is 7 days.

[0062] (1) Sleep test experiment:

[0063] After continuous administration for 28 days, the sleep situation of the mice is observed. The disappearance time of the righting reflex is recorded. The righting reflex disappears when the mouse is in the dorsal recumbent position and cannot right itself within 60 s. The time from the disappearance of the righting reflex to its recovery is the sleep time of the mouse. The number of mice falling asleep and the sleep time in each group are observed and recorded respectively, as shown in Table 2.

[0064] Table 2

[0065]

[0066] After 28 days of the experiment, statistics on the sleep conditions of the mice after 5 minutes, 10 minutes, and 15 minutes showed that the sleep-aid composition prepared from the raw materials, ratio, and production method of the present invention was more likely to induce sleep after administration, and the duration of continuous sleep increased.

[0067] (II) Brain tissue experiment:

[0068] In this study, the contents of 5-HT, GABA, and DA in the brain tissues of mice were measured, as shown in Table 3. 5-HT is a monoamine neurotransmitter involved in sleep regulation, and an increase in the content of 5-HT can improve insomnia symptoms; GABA is the main inhibitory neurotransmitter in the brain tissue, distributed in parts such as the cerebral cortex, hypothalamus, brainstem, hippocampus, and basal ganglia. The release of GABA in the cortex increases during sleep, and activating the GABA receptor can promote sleep; DA plays an important role in the body's motor control, reward, mood regulation, and addictive behaviors. When the content of DA is too high, the body will have a sense of excitement, leading to insomnia.

[0069] Table 3

[0070]

[0071] The results of the brain tissue experiment of the mice showed that after continuously administering the sleep-aid composition of the embodiment of the present invention for 28 days, the contents of 5-HT and GABA in the brain tissues of the mice could be increased, and the content of DA could be decreased. Compared with Comparative Examples 1-4, the 5-HT value in Example 1 increased by 3.2 pg / mg, GABA increased by 186.7 pg / mg, and DA decreased by 0.55 pg / mg.

[0072] Conclusion: The sleep-aid composition described in the present invention has a significant effect on regulating sleep. After continuously taking it for 28 days, the sleep-aid effects of Examples 1-3 are higher than those of the comparative examples, positive control, and blank control. The production process of the wild ganoderma lucidum extract of the present invention has a better effect. In Comparative Example 1, the filtration step of the wild ganoderma lucidum extract was changed, and an ultrafiltration membrane was not used. After filtering with filter paper, the increase in the contents of 5-HT and GABA and the decrease in the value of DA in the brain tissues of the mice were all lower than those in Example 1. In Comparative Example 2, when the component content of the sephadex gel column was changed during the extraction of the wild ganoderma lucidum extract, the sleep onset time of the mice increased, and the contents of 5-HT and GABA in the brain tissues were lower. After removing selenium-enriched yeast from the raw materials in Comparative Example 3, the sleep onset time of the mice increased, the contents of 5-HT and GABA in the brain tissues were lower, and the DA value was higher. It shows that the compounding effect of the sleep-aid composition prepared from the raw materials, ratio, and production method of the present invention is better, and it has a significant curative effect in terms of helping sleep.

[0073] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A method for preparing a tranquilizing and sleep-inducing composition, characterized in that: The composition is composed of 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 composition for calming nerves and aiding sleep comprises the following preparation steps: weighing wild ganoderma lucidum extract and wild cordyceps sinensis, mixing, sieving, concentrating, adding β-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; 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, passing through a dextran gel column, filtering through an ultrafiltration membrane, concentrating and drying the supernatant at 60-65° C. to obtain wild ganoderma lucidum powder, and storing at low temperature; The ultrasonic low-oxygen extraction has an extraction temperature of 30-45°C, an extraction time of 25-30 minutes, a standing time of 10-15 minutes, a frequency of 20-35kHz, and an oxygen content of less than 5% during extraction; The components of the dextran gel column equilibrium solution are: 20-23mM Hepes-NaOH, 1.5-2mM Na2-EDTA, 4-6mM MgCl2, pH 7.5-8.

2. The method for preparing a tranquilizing and sleeping-aiding composition according to claim 1, characterized in that: In the preparation steps of the tranquilizing and sleep-aiding composition, the sieving is performed through 200-240 meshes, and the concentration is performed, and the relative density of the concentrated solution 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: In the preparation of wild ganoderma lucidum extract, the molecular weight cutoff of ultrafiltration membrane filtration is 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.

6. Use of the composition prepared by the preparation method according to any one of claims 1 to 5 in preparing a drug for treating insomnia.

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