CN113826896A

CN113826896A - Health food for improving sleep and preparation method thereof

Info

Publication number: CN113826896A
Application number: CN202111113471.XA
Authority: CN
Inventors: 陈耀明; 陈伟; 李顺兰
Original assignee: Nutryfarm Chengdu Biomedicine Co ltd
Current assignee: Nutryfarm Chengdu Biomedicine Co ltd
Priority date: 2021-09-23
Filing date: 2021-09-23
Publication date: 2021-12-24

Abstract

The invention provides a health food for improving sleep and a preparation method thereof, and the health food comprises the following raw materials in percentage by mass: 15-30% of spina date seed extract, 10-20% of fructus schizandrae powder, 10-18% of poria cocos extract and 10-18% of lily extract. Animal experiments prove that the health food is safe, has no toxic or side effect, is suitable for long-term administration, and has the function of improving sleep.

Description

Health food for improving sleep and preparation method thereof

Technical Field

The invention relates to the technical field of health-care food, in particular to health-care food for improving sleep and a preparation method thereof.

Background

With the aggravation of social competition and the acceleration of life rhythm, the life pressure of people is increased, and the incidence rate of sleep disorder is on the rise. Sleep disorders are characterized by abnormal amounts of sleep and abnormal behavior in sleep, as well as by disturbances in the regular rhythmic alternation of sleep and arousal. Insomnia is a common sleep disorder. Sleep disorders such as insomnia and the like can bring consequences such as anxiety, depression, attention deficit, dysmnesia and the like to people, cause problems such as physical and mental fatigue, general immunologic function reduction and the like, and bring serious harm to the body and mind of patients.

Therefore, improving sleep quality is of great significance to prevent diseases and protect physical and mental health of people. At present, methods for improving sleep and treating sleep disorders such as insomnia mainly include methods for adjusting life rhythm, properly exercising, psychologic dispersion, food conditioning, drug therapy and the like. The medicine treatment effect is good, but the medicine treatment usually needs to be taken for a long time, so that the medicine is easy to depend on, and rebound is easy to happen after the medicine is stopped; and the medicaments for treating the sleep disorder are mainly alprazolam, estazolam, clozapine, lorazepam, zolpidem, zopiclone and the like, and have toxic and side effects after being taken for a long time, so that the burden of liver and kidney functions is increased, the gastrointestinal tract is stimulated, and the normal digestive function is influenced. For most patients with mild symptoms, food can be prepared to achieve ideal effects, so that the health-care food provides ideal choices for the patients with mild symptoms.

However, although there are many kinds of health foods on the market, the specific symptoms are not targeted, and the types of health foods specifically used for improving sleep quality are few and the effects are not uniform. Therefore, the invention develops the health food for improving the sleep quality, which is eaten by patients with sleep disorder.

Disclosure of Invention

Aiming at the defects of the prior art, the invention aims to provide the sleep-improving health food which is strong in pertinence, good in effect and suitable for long-term taking and the preparation method thereof.

The invention is realized by the following technical scheme:

a health food for improving sleep comprises the following raw materials in percentage by mass: 15-30% of spina date seed extract, 10-20% of fructus schizandrae powder, 10-18% of poria cocos extract and 10-18% of lily extract.

In one embodiment of the present application, the raw materials comprise, in mass percent: 22-25% of spina date seed extract, 15-20% of fructus schizandrae powder, 13-15% of poria cocos extract and 13-15% of lily extract.

In one embodiment of the present application, the spine date seed extract is prepared by: mashing cleaned semen Ziziphi Spinosae, soaking in water, decocting, filtering, concentrating into paste, and pulverizing the dry extract to obtain semen Ziziphi Spinosae extract powder; and/or

The schisandra chinensis powder is prepared by the following steps: taking clean schisandra chinensis, and adding the mixture into the mixture according to a solid-liquid mass ratio of 1: 2-5, soaking, heating and cooking until the surface is black purple or brown, taking out, drying and crushing to obtain schisandra chinensis powder; and/or

The poria cocos extract and the lily extract are prepared in the following modes: respectively taking clean poria cocos and clean lily, crushing, heating and water leaching powder, concentrating to obtain concentrated solution, adding 95% ethanol into the concentrated solution for precipitation, standing, centrifuging to obtain precipitate, washing the precipitate with absolute ethanol, acetone or diethyl ether, and drying the washed precipitate in vacuum to obtain the poria cocos extract and the lily extract.

In an embodiment of the present application, the raw material composition and mass fraction of the health food further include: 3% -5% of wall-broken ganoderma lucidum spore powder and/or 5% -10% of gastrodia elata extract.

In one embodiment of the present application, the gastrodia elata extract is prepared by: crushing the tuber of the clean gastrodia elata, adding ethanol to soak for 45-60 min, heating and continuously refluxing for 30-45 min in a Soxhlet extractor, cooling, centrifuging, taking supernatant, concentrating and drying to obtain the gastrodia elata extract.

In an embodiment of the present application, the raw materials comprise, in mass percent: 25% of spina date seed extract, 20% of Chinese magnoliavine fruit powder, 15% of poria cocos extract, 15% of lily extract, 5% of wall-broken ganoderma lucidum spore powder and the balance of corn starch.

In an embodiment of the present application, the raw materials comprise, in mass percent: 30% of spina date seed extract, 10% of Chinese magnoliavine fruit powder, 10% of poria cocos extract, 10% of lily extract, 3% of wall-broken ganoderma lucidum spore powder, 7% of gastrodia elata extract and the balance of corn starch.

In one embodiment of the present application, the total saponin content of the health food is more than or equal to 0.38%.

A preparation method of a health food for improving sleep comprises the following steps:

step S1, respectively pulverizing the spina date seed extract, the Chinese magnoliavine fruit powder, the poria cocos extract and the lily extract, and sieving the pulverized materials with a 80-mesh sieve to prepare fine powder for later use;

step S2, respectively weighing the spine date seed extract, the Chinese magnoliavine fruit powder, the poria cocos extract and the lily extract fine powder prepared in the step S1, and mixing the spine date seed extract, the Chinese magnoliavine fruit powder, the poria cocos extract and the lily extract fine powder with corn starch sieved by a 80-mesh sieve to prepare mixed powder;

and step S3, filling the mixed powder into capsules to obtain capsules with filling quantity of 400 mg/capsule.

step S1, respectively pulverizing the spina date seed extract, the Chinese magnoliavine fruit powder, the poria cocos extract, the lily extract, the wall-broken ganoderma lucidum spore powder and the gastrodia elata extract, and sieving the pulverized powder with a 80-mesh sieve to prepare fine powder for later use;

step S2, respectively weighing the spina date seed extract, the Chinese magnoliavine fruit powder, the poria cocos extract, the lily extract, the wall-broken ganoderma lucidum spore powder and the fine gastrodia tuber extract powder prepared in the step S1, and mixing the fine spina date seed extract, the Chinese magnoliavine fruit powder, the poria cocos extract, the lily extract, the wall-broken ganoderma lucidum spore powder and the fine gastrodia tuber extract powder with corn starch sieved by a 80-mesh sieve to prepare mixed powder;

Compared with the prior art, this beneficial effect is:

the health food provided by the invention is developed according to the traditional Chinese medicine theory and the modern medical theory, takes the spina date seed extract as the main material, takes the Chinese magnoliavine fruit powder as the auxiliary material, and is matched with the poria cocos extract, the lily extract and the like, so that the health food has the effects of calming the heart, arresting sweating, promoting the production of body fluid, tonifying qi and kidney, improving insomnia, dysphoria with pavor, mental fatigue and the like, and can be eaten by adults with poor sleep; the health food has mild drug property, can condition viscera, regulate yin and yang balance, and improve sleep, has no incompatibility between raw materials, is safe for eating, has exact sleep improving function, and is suitable for long-term administration.

Detailed Description

Example 1

A health food for improving sleep comprises the following raw materials in parts by mass: 25% of spina date seed extract, 20% of Chinese magnoliavine fruit powder, 15% of poria cocos extract, 15% of lily extract and 25% of corn starch.

The preparation method of the spina date seed extract comprises the following steps: taking dry and clean spina date seeds, mashing, adding distilled water according to the solid-liquid mass ratio of 1:10, soaking for 8 hours, decocting, boiling for 1 hour, filtering, and keeping filtrate; adding water into filter residues for secondary decoction, boiling for 1-2 hours, and filtering; mixing the filtrates, heating, and concentrating to obtain extract; drying the concentrated extract, and pulverizing to obtain semen Ziziphi Spinosae extract powder.

The preparation method of the schisandra chinensis powder comprises the following steps: taking dry and clean schisandra chinensis, mashing, and adding a mixture of schisandra chinensis and water in a solid-liquid ratio of 1: 3, sealing and soaking, heating in a steamer, fumigating with yellow wine steam or directly steaming until the surface of fructus Schisandrae chinensis is black purple or brown, taking out, drying, and pulverizing to obtain fructus Schisandrae chinensis powder.

The preparation method of the tuckahoe extract comprises the following steps: weighing clean and dry diced or blocky poria cocos, crushing, adding the powder into hot water for leaching, wherein the solid-to-liquid ratio is 1: 30, leaching for 3 hours at the hot water temperature of 85 ℃; and carrying out suction filtration on the leaching liquor to obtain filtrate, carrying out vacuum decompression concentration on the filtrate to obtain concentrated solution, wherein the volume ratio of the filtrate to the concentrated solution is 10: 1; adding 10 times volume of 95% ethanol into the concentrated solution for precipitation to ensure that the alcohol content of the precipitation solution is over 85%, standing at 4 deg.C for 8 hr, and centrifuging to obtain precipitate; washing the precipitate with anhydrous alcohol, acetone or diethyl ether, and vacuum drying the washed precipitate to obtain Poria extract.

The obtained Poria extract contains pachyman as main ingredient.

The preparation method of the lily extract comprises the following steps: weighing clean and dry lily, crushing, adding hot water for leaching, wherein the solid-to-liquid ratio is 1: 40, leaching for 8 hours at the hot water temperature of 70 ℃; and carrying out suction filtration on the leaching liquor to obtain filtrate, carrying out vacuum decompression concentration on the filtrate to obtain concentrated solution, wherein the volume ratio of the filtrate to the concentrated solution is 10: 1; adding 95% ethanol into the concentrated solution for precipitation, wherein the volume ratio of the 95% ethanol to the concentrated solution is 10: 3, ensuring that the alcohol content of the precipitation solution is more than 70%, standing at the low temperature of 4 ℃ for 8 hours, and centrifuging to obtain a precipitate; washing the precipitate with anhydrous ethanol, and vacuum drying the washed precipitate to obtain Bulbus Lilii extract.

The Bulbus Lilii extract obtained by this method contains Bulbus Lilii polysaccharide as main ingredient.

The preparation method of the health food for improving sleep comprises the following steps:

step S1, respectively pulverizing the spine date seed extract, the Chinese magnoliavine fruit powder, the poria cocos extract and the lily extract, and sieving the pulverized materials with a 80-mesh sieve to prepare fine powder for later use; the corn starch is also sieved by a 80-mesh sieve and prepared into fine powder.

Step S2, weighing 2.5kg of wild jujube seed extract, 2.0kg of Chinese magnoliavine fruit powder, 1.5kg of tuckahoe extract, 1.5kg of lily extract and 2.5kg of corn starch respectively according to the formula proportion, and mixing for 30min by using a three-dimensional mixer (specification: SYH-200) to obtain mixed powder.

And step S3, filling the prepared mixed powder into capsules by using a full-automatic capsule filling machine (specification model: NJP-2000 c) to prepare capsule granules with the filling quantity of 400 mg/granule.

And step S4, polishing to remove powder on the surface of the capsule, and selecting and removing unqualified capsule particles.

And step S5, filling the oral solid medicinal polyester bottles meeting the standard, wherein the specification is 60 granules/bottle.

And step S6, carrying out external packaging, then inspecting the finished product, and warehousing after the finished product is qualified.

The above steps S1 to S5 are performed in a production plant having a cleanliness of thirty thousand levels.

The corn starch adopted in the embodiment can be replaced by other edible solid auxiliary materials.

The content of total saponins in the health food prepared in the embodiment is about 0.40%.

Example 2

A health food for improving sleep comprises the following raw materials in parts by mass: 25% of spina date seed extract, 20% of Chinese magnoliavine fruit powder, 15% of poria cocos extract, 15% of lily extract, 5% of wall-broken ganoderma lucidum seed powder and 20% of corn starch.

Wherein, the spina date seed extract, the schisandra chinensis powder, the poria cocos extract and the lily extract are prepared by adopting the preparation method corresponding to the embodiment 1.

The preparation method of the wall-broken ganoderma lucidum spore powder comprises the following steps: weighing 200g of mature and plump ganoderma lucidum spore powder, adding 800g of corn starch, 0.1g of cellulase (with the enzyme activity of 20 ten thousand U/g), 0.1g of chitinase (with the enzyme activity of 10 ten thousand U/g) and 0.05g of glucosidase (with the enzyme activity of 20 ten thousand U/g), adding the mixture into warm water for fully mixing, and soaking and fermenting the mixture for 4 to 6 hours at 40 ℃ in a closed container to soften and degrade the ganoderma lucidum spore wall; then rapidly heating the fermented product to 70 ℃, maintaining for 15 minutes, pasting the corn starch, breaking the wall of the ganoderma lucidum spore powder, and simultaneously inactivating the cellulase, the chitinase and the glucosidase; and finally drying and crushing the fermentation product to obtain the wall-broken ganoderma lucidum spore powder.

step S1, respectively pulverizing the spina date seed extract, the Chinese magnoliavine fruit powder, the poria cocos extract, the lily extract and the wall-broken ganoderma lucidum spore powder, and sieving the pulverized powder with a 80-mesh sieve to prepare fine powder for later use; the corn starch is also sieved by a 80-mesh sieve and prepared into fine powder.

Step S2, weighing 2.5kg of spina date seed extract, 2.0kg of Chinese magnoliavine fruit powder, 1.5kg of poria cocos extract, 1.5kg of lily extract, 0.5kg of wall-broken ganoderma lucidum spore powder and 2.0kg of corn starch respectively according to the formula proportion, and mixing for 30min by using a three-dimensional mixer (specification model: SYH-200) to obtain mixed powder.

The content of total saponins in the health food prepared in the embodiment is about 0.41%.

Example 3

A health food for improving sleep comprises the following raw materials in parts by mass: 30% of spina date seed extract, 10% of Chinese magnoliavine fruit powder, 10% of poria cocos extract, 10% of lily extract, 3% of wall-broken ganoderma lucidum pao powder, 7% of gastrodia elata extract and 30% of corn starch.

Wherein, the spina date seed extract, the schisandra chinensis powder, the poria cocos extract and the lily extract are prepared by the corresponding preparation method in the embodiment 1. Sporoderm-broken ganoderma lucidum spore powder was prepared using the preparation method described in example 2.

The preparation method of the rhizoma gastrodiae extract comprises the following steps: weighing dry and clean rhizoma gastrodiae tubers, crushing, and adding the powder into a container with the volume ratio of 1: soaking in 10% 75% ethanol for 45min, heating in Soxhlet extractor, reflux extracting for 45min, cooling, centrifuging, collecting supernatant, concentrating under reduced pressure, recovering ethanol, concentrating to 1-2 g/ml crude drug, adding corn starch, mixing, and vacuum drying to obtain rhizoma Gastrodiae extract.

The rhizoma Gastrodiae extract obtained by this method contains gastrodine as main ingredient.

step S1, respectively pulverizing the spina date seed extract, the Chinese magnoliavine fruit powder, the poria cocos extract, the lily extract, the wall-broken ganoderma lucidum spore powder and the gastrodia elata extract, and sieving the pulverized powder with a 80-mesh sieve to prepare fine powder for later use; the corn starch is also sieved by a 80-mesh sieve and prepared into fine powder.

And step S2, weighing 6.0kg of spina date seed extract, 2.0kg of Chinese magnoliavine fruit powder, 2.0kg of poria cocos extract, 2.0kg of lily extract, 0.6kg of wall-broken ganoderma lucidum spore powder, 1.4kg of gastrodia elata extract and 6.0kg of corn starch respectively according to the formula proportion, and mixing for 30min by using a three-dimensional mixer (specification model: SYH-200) to obtain mixed powder.

The content of total saponins in the health food prepared in the embodiment is about 0.39%.

Example 4

A health food for improving sleep comprises the following raw materials in parts by mass: 22% of spina date seed extract, 15% of Chinese magnoliavine fruit powder, 13% of poria cocos extract, 15% of lily extract, 10% of gastrodia elata extract and 25% of corn starch.

Wherein, the spina date seed extract, the schisandra chinensis powder, the poria cocos extract and the lily extract are prepared by adopting the preparation method corresponding to the embodiment 1; the wall-broken ganoderma lucidum spore powder is prepared by the corresponding preparation method in the embodiment 2; the gastrodia elata extract was prepared according to the preparation method described in example 3.

step S1, respectively pulverizing the spina date seed extract, the Chinese magnoliavine fruit powder, the poria cocos extract, the lily extract and the gastrodia elata extract, and sieving the pulverized materials with a 80-mesh sieve to prepare fine powder for later use; the corn starch is also sieved by a 80-mesh sieve and prepared into fine powder.

And step S2, weighing 4.4kg of spina date seed extract, 3.0kg of Chinese magnoliavine fruit powder, 2.6kg of poria cocos extract, 3.0kg of lily extract, 2.0kg of gastrodia elata extract and 5.0kg of corn starch respectively according to the formula proportion, and mixing for 40min by using a three-dimensional mixer (specification model: SYH-200) to prepare mixed powder.

Example 4

Example 5

A health food for improving sleep comprises the following raw materials in parts by mass: 15% of spina date seed extract, 20% of Chinese magnoliavine fruit powder, 18% of poria extract, 18% of lily extract, 4% of wall-broken ganoderma lucidum spore powder, 5% of gastrodia elata extract and 20% of corn starch.

And step S2, weighing 1.5kg of spina date seed extract, 2.0kg of Chinese magnoliavine fruit powder, 1.8kg of poria cocos extract, 1.8kg of lily extract, 0.4kg of wall-broken ganoderma lucidum spore powder, 0.5kg of gastrodia elata extract and 2.0kg of corn starch respectively according to the formula proportion, and mixing for 35min by using a three-dimensional mixer (specification model: SYH-200) to obtain mixed powder.

The content of total saponins in the health food prepared in the embodiment is about 0.42%.

Example 6

Safety toxicology evaluation test

First, acute oral toxicity test (maximum tolerated dose method, i.e. MTD method)

The test substance was the capsule preparation prepared in example 5, the content was brown powder, and the recommended dose for adults was 800 mg/day, and the test substance was dissolved in distilled water to prepare the desired concentration.

SD rats (provided by WUDUDOU Biotechnology Co., Ltd., certification number: SCXK (Chuan) 2008-24) 20 rats with weight of 180-220 g and half of male and female. Setting a dosage group of 10 g/kg.bw, weighing 50g of the test object, preparing distilled water to 100 ml, performing oral gavage once according to 2ml/100g.bw, observing the number of dead animals and general health conditions after the gavage, and judging the acute toxicity of the test object according to the maximum tolerance.

The test results of the acute oral toxicity test are shown in table 1, and after the test object is subjected to oral toxicity at a dose of 10 g/kg.bw, no animal death is seen within two weeks, and no obvious toxic symptoms or adverse reactions are caused. After the two-week experiment, all animals were killed and no obvious abnormal change of internal organs was observed.

And (4) conclusion: the test substance has acute oral MTD of more than 10 g/kg.bw for female and male SD rats, and is practically nontoxic.

Second, genotoxicity test

1. Contaminant mutagenicity detection assay (Ames assay)

Standard plate-entry tests (0.5 ml of S9 mixture per plate) with and without enlargement of rat liver S9 (induced by polychlorinated biphenyl) were performed using the identified satisfactory strains TA97, TA98, TA100 and TA 102.

Five subject doses were set: 8. 40, 200, 1000 and 5000. mu.g/dish. 1.0g of the test substance is accurately weighed and distilled water is added to 20ml to obtain the maximum working concentration of 50 mg/ml. 100 μ l was added to the plate to a maximum final concentration of 5000 μ g/plate. The rest working concentration is obtained by diluting with distilled water 5 times. All prepared working solutions were autoclaved at 8 lbs for 15 min. 100 μ l of each sample was added to the plate to obtain the desired concentration.

A spontaneous control, a solvent control (100. mu.l/dish of sterilized distilled water) and a positive control were set. The positive control without S9 test was 2,4, 7-trinitrofluorene ketone (2, 4, 7-TNFanone) (0.2. mu.g/dish, applicable strains TA97, TA 98); sodium azide (NaN)₃) (1.5. mu.g/dish, applicable strain TA 100), mitomycin C (MMC) (0.5. mu.g/dish, applicable strain TA 102); the positive control for the addition of S9 was 2-aminofluorene (2-AF) (10.0. mu.g/dish, adapted strain TA97, TA98, TA 100) and 1, 8-dihydroxyanthraquinone (Dan) (50.0. mu.g/dish, adapted strain TA 102). Three parallel dishes were made for each test dose and the test was repeated once.

The test results of the Ames test are shown in tables 2 and 3, and the spontaneous control group showed an obvious positive reaction with the average number of colonies transformed back per dish within the normal range, and the positive control group induced the average number of colonies transformed back per dish more than twice of the spontaneous control group, regardless of the test with or without the addition of S9. The average number of the reversion colonies of each dose group of the test object does not exceed one time of that of the solvent control group, and the test object shows a negative reaction.

And (4) conclusion: the test substance has no effect of inducing the test strain to carry out back mutation.

2. Micronucleus test for myelophilis pleochromocytes

SPK-grade KM mice (provided by Sichuan academy of traditional Chinese medicine) are 25 g-30 g in weight, and are randomly divided into 5 groups of 10 mice each with half of male and female.

Negative control (distilled water), positive control (cyclophosphamide 40 mg/kg. bw) and 2.5, 5.0 and 10.0 g/kg. bw test groups of test substance were set up (5.0 g, 10.0g and 20.0g of test substance, respectively, were weighed, and distilled water was added to 40 ml).

Animals were gavaged at 0 h and 24 h at 2ml/100g.bw orally, sacrificed 6 h after the last exposure, and processed into tablets.

Counting 1000 Pleochromocyte (PCF) cells in each animal, observing the number of the Pleochromocyte (PCF) cells containing micronucleus, calculating the micronucleus rate (‰), simultaneously observing the number of PCE and RBC in 200 red blood cells, and calculating the PCE/RBC ratio.

The result of the mouse bone marrow pleochromocyte micronucleus test is shown in table 4, the micronucleus rates of the female and male animals in the positive control group are both obviously higher than that in the negative control group (P is less than 0.01), and the micronucleus rates of all groups of the tested object have no significant difference (P is more than 0.05) compared with that in the negative control group, which indicates that the tested object is a negative result in the mouse bone marrow pleochromocyte micronucleus test.

3. Mouse teratospermia test

SPK-grade KM male mice (provided by Sichuan academy of traditional Chinese medicine, State of health: SCXK 2008-19) weigh 25 g-35 g and are randomly divided into 5 groups of 5 mice.

The test is carried out by three test substance dosage groups of a negative control group and a positive control group (cyclophosphamide CP 40 mg/kg.bw) as well as 2.5 g/kg.bw, 5.0 g/kg.bw and 10.0 g/kg.bw (the same micronucleus test is carried out by preparing gastric lavage liquid), and the animals are orally gazed according to 2ml/100g.bw for 5 days continuously. Animals were sacrificed 35 days after the first gavage, bilateral epididymis were removed and processed according to standard procedures, 1000 intact sperm per animal were observed, and the number of teratospermis was recorded.

The mouse teratospermia test result is shown in table 5, the sperm teratogenesis rate of the positive control group is significantly higher than that of the negative control group (P is less than 0.01), and the sperm teratogenesis rate of each dosage group of the test object has no significant difference (P is more than 0.05) compared with that of the negative control group, which indicates that the test object is a negative result in the mouse teratospermia test.

In conclusion, Ames test, mouse marrow pleochromocyte micronucleus test and mouse sperm malformation test are all negative, which indicates that the test substance has no mutagenic effect.

Three, rat 30 days feeding test

The test substance was the capsule preparation prepared in example 5, and the content was brown powder, and the recommended dose for adults was 800 mg/day.

Weaning SD rats (provided by Sichuan academy of traditional Chinese medicine) were divided into four groups of 20 rats each with half each female and half each group by body weight after one week of adaptive feeding.

The test was set up with one negative control group (basal feed) and three test dose groups: bw of 400 mg/kg, 1000 mg/kg, bw and 1600mg/kg, bw (equivalent to 30, 75 and 120 times the recommended intake for humans). The test substance is mixed into the feed according to 10% of the animal weight. The preparation method of the feed comprises the following steps: 4.0g, 10g and 16g of the test substances are respectively added into each 1kg of feed, and the materials are processed into granulated feed and fed for 30 days.

Animal body weight, feed intake and food remaining were weighed once a week and food intake calculated. The weekly food utilization (%) was calculated as weekly weight gain and weekly food intake, respectively, and the total food utilization (%) was calculated at the same time. Animals were sacrificed after 30 days and hematological, sero-biochemical and histopathological examination and organ coefficient measurements were performed.

Animals in the negative control group and 3 dose groups had normal food intake, water drinking, stool and urine, good growth and development conditions and general performance during the feeding period of 30 days, and no obvious behavior change and toxic performance were observed.

(1) Influence on body weight and food availability

The body weight and food availability results for each experimental group over the entire test period are shown in tables 6 and 7. The weight, total weight gain and total food utilization rate of animals in each week of each dose group of the test object are not significantly different (P is more than 0.05) compared with the negative control group through variance analysis, and the test object has no adverse effect on the growth and development of organisms.

(2) Organ coefficient measurement results

As shown in table 8, the weights and organ coefficients (organ wet weight/body weight × 100) of the important organs in each dose group of the test substances were shown, and it was found from table 8 that the weights and organ coefficients of the important organs in each dose group of the female and male mice were not significantly different from those in the negative control group (P > 0.05).

(3) End-stage hematological test results

As shown in Table 9, the results of measurement of the conventional hematological indices (red blood cells, hemoglobin, white blood cells, and classification) at the end of the test are shown. The hematology determination indexes of female and male mice of each dose group of the test object have no significant difference compared with a negative control group (P is more than 0.05).

(4) End stage biochemical index test result

As shown in Table 10, the results of the biochemical tests of liver function, kidney function, blood lipid and blood glucose in each experimental group show that the indexes of each dose group have no significant difference (P >0.05) compared with the negative control group.

(5) Histological examination results

Gross and histopathological examination of liver, kidney, stomach, jejunum, spleen, ovary/testis, etc. was performed at the end of the experiment. Gross examination revealed no significant lesions, so only the high dose and negative control groups were selected for histological examination.

See tables 11-17, see under the mirror: one example of mild chronic pyelonephritis is found in the control group and the high-dose group, and the lesions are comprehensively analyzed and belong to spontaneous lesions of animals. Therefore, no lesion change due to the test substance was observed in the organs of the animals in each dose group.

In summary, the results of the biological indicators in tables 6 to 17 show that the test substance does not cause abnormal changes in important indicators such as the overall health condition, physiological and biochemical functions and organ morphology of the rat in the 30-day feeding test of the rat with the highest dose 120 times (1600 mg/kg.bw) of the recommended intake of the human body.

To summarize: the sleep-improving health food obtained in example 5 had an acute oral MTD of more than 10 g/kg.bw for SD rats, and was practically non-toxic. The Ames test, the mouse bone marrow cell micronucleus test and the mouse teratospermia test are all negative results, which indicate that the test substance has no mutagenic effect. In a 30-day feeding test of rats with the highest dose 120 times (1600 mg/kg. bw) of the recommended intake of human bodies, the maximum non-effective dose of the test object is preliminarily estimated to be more than 1600mg/kg.bw without causing abnormal changes of important indexes of the overall health condition, physiological and biochemical functions and organ tissue morphology of the rats.

Therefore, the health food for improving sleep can be effectively proved to be eaten for a long time, safe and free of toxic and side effects.

Example 7

Sleep improvement test

The test substance was the capsule preparation prepared in example 1, the content was brown powder, and the recommended adult dose was 800 mg/day, and the gastric lavage fluid was prepared with distilled water.

SPF-grade female ICR mice (provided by Sichuan Biotechnology, Inc.) were randomly divided into 4 groups, i.e., negative control group and three dose groups of test substance 0.13 g/kg.bw, 0.27 g/kg.bw and 0.40g/kg.bw (corresponding to 10, 20 and 30 times of recommended intake of human body, respectively). The contents of the test capsule, 0.26g, 0.54g and 0.80g, were dissolved in 40ml of distilled water (prepared daily) and used as gastric perfusion fluid in the three dosage groups of high, medium and low. Gavage was performed once a day for 30 consecutive days at 2% volume. The negative control group was gavaged with an equal volume of distilled water.

1. Direct sleep test

The animals were observed for the presence of sleep (sleep indicated by disappearance of the righting reflex) after the administration of the test samples.

As a result: the animals in each group did not sleep after the test sample was given, indicating that the test sample had no direct sleep.

2. Test for prolonging sleep time of sodium pentobarbital

After 30 minutes of the last gastric lavage of the test sample or the swallow distilled water, 45 mg/kg. bw of sodium pentobarbital is injected into the abdominal cavity of each group of animals, and the duration of sleep (i.e. loss of righting reflex) of the animals after the injection is observed and recorded. The average sleeping time of each group of animals was calculated separately.

As shown in Table 18, the average sleeping time of the mice in the three dose groups of the test object is longer than that of the control group, but the average sleeping time is not statistically significant (P >0.05), which indicates that the test object has no significant prolonging effect on the sleeping time of the mice induced by the pentobarbital sodium.

3. Pentobarbital sodium subthreshold dose hypnosis test

After 30 minutes of the last gastric lavage of the test sample or distilled water, 27 mg/kg. bw of sodium pentobarbital is injected into the abdominal cavity of each group of animals, and the number of the animals falling asleep within 30 minutes after the injection is observed (the standard for judging the falling asleep is that the disappearance of the righting reflex exceeds 1 minute). The incidence of sleep was calculated for each group of animals separately.

As shown in Table 19, the incidence of sleep in mice of each dose group of the test substance is higher than that of the control group, and the incidence of sleep in mice of the high dose group is remarkably increased (P is less than 0.05) compared with that of the control group, which indicates that the test substance can increase the incidence of sleep in mice induced by low dose sodium pentobarbital.

4. Barbiturate sodium sleep latency test

After 30 minutes of the last gastric lavage of the test sample or distilled water, 240 mg/kg. bw of barbital sodium is injected into the abdominal cavity of each group of animals, and the time of falling asleep of the animals after injection is observed and recorded. The mean sleep latency (i.e. time to fall asleep-barbiean sodium injection time) was calculated for each group of animals.

As shown in table 20, the average sleep latency of mice in each dose group of the test substance is reduced compared with the control group, and the average sleep latency of mice in the high dose group is significantly reduced compared with the control group (P < 0.05), which indicates that the test substance can shorten the sleep latency of mice induced by barbiturate.

In conclusion, the test result of the test object for improving the sleep function shows that the test object has no direct sleep effect on the mouse; the 0.40g/kg. bw dosage group mice had significantly higher sleep onset rates than the control group and significantly shorter average sleep latency than the control group. The test substance has the function of improving sleep according to evaluation criteria.

Claims

1. The health food for improving sleep is characterized by comprising the following raw materials in percentage by mass: 15-30% of spina date seed extract, 10-20% of fructus schizandrae powder, 10-18% of poria cocos extract and 10-18% of lily extract.

2. The sleep-improving health food as claimed in claim 1, wherein the raw materials comprise, in mass percent: 22-25% of spina date seed extract, 15-20% of fructus schizandrae powder, 13-15% of poria cocos extract and 13-15% of lily extract.

3. The sleep-improving health food as claimed in claim 1, wherein the spine date seed extract is prepared by: mashing cleaned semen Ziziphi Spinosae, soaking in water, decocting, filtering, concentrating into paste, and pulverizing the dry extract to obtain semen Ziziphi Spinosae extract powder; and/or

4. The sleep-improving health food as claimed in claim 1 or 3, wherein the raw material composition and mass fraction of the health food further comprises: 3% -5% of wall-broken ganoderma lucidum spore powder and/or 5% -10% of gastrodia elata extract.

5. The sleep-improving health food as claimed in claim 4, wherein the Gastrodia elata extract is prepared by: crushing the tuber of the clean gastrodia elata, adding ethanol to soak for 45-60 min, heating and continuously refluxing for 30-45 min in a Soxhlet extractor, cooling, centrifuging, taking supernatant, concentrating and drying to obtain the gastrodia elata extract.

6. The sleep-improving health food as claimed in claim 4, wherein the raw materials comprise, in mass percent: 25% of spina date seed extract, 20% of Chinese magnoliavine fruit powder, 15% of poria cocos extract, 15% of lily extract, 5% of wall-broken ganoderma lucidum spore powder and the balance of corn starch.

7. The sleep-improving health food as claimed in claim 4, wherein the raw materials comprise, in mass percent: 30% of spina date seed extract, 10% of Chinese magnoliavine fruit powder, 10% of poria cocos extract, 10% of lily extract, 3% of wall-broken ganoderma lucidum spore powder, 7% of gastrodia elata extract and the balance of corn starch.

8. The health food for improving sleep according to any one of claims 1, 2, 6 and 7, wherein the total saponin content of the health food is not less than 0.38%.

9. The method for preparing a health food for improving sleep as set forth in any one of claims 1 to 3, comprising the steps of:

10. The method for preparing a health food for improving sleep as set forth in claim 7, comprising the steps of: