CN104825574B - A kind of composition for improving sleep and preparation method thereof - Google Patents
A kind of composition for improving sleep and preparation method thereof Download PDFInfo
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- CN104825574B CN104825574B CN201510172815.2A CN201510172815A CN104825574B CN 104825574 B CN104825574 B CN 104825574B CN 201510172815 A CN201510172815 A CN 201510172815A CN 104825574 B CN104825574 B CN 104825574B
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Abstract
The present invention relates to health product technology field, more particularly to a kind of composition for improving sleep and preparation method thereof.The present invention can have the function of being significantly improved sleep after being compounded apple stem cell freeze-dried powder, epiphysin and vitamin B6, and have no toxic side effect.Experiment shows that the significant effect of the improvement sleep of composition provided by the invention shows that mouse does not note abnormalities through acute toxicity test in mice result, poisoning or the phenomena of mortality do not occur better than the effect that each component is used alone.And composition provided by the invention can effectively improve yellow Jackets sub-threshold dose mouse sleep incidence under 0.5g/kg dosage.
Description
Technical field
The present invention relates to health product technology field, more particularly to a kind of composition for improving sleep and preparation method thereof.
Background technology
Modern allegro life and gradually violent various competitive relations, make people's stress increasing, spirit
Long-term nervous and a series of irregular rhythm of life, the insomnia caused annoyings most people always.According to statistics, China
Insomniac is the 20%~30% of total population, and wherein the elderly accounts for 70%.Chronic insomnia easily causes vexed easily random, tired nothing
Power, or even with headache, dreaminess, hidrosis, failure of memory, can also cause a series of clinical symptoms, and induce some psychosomatic diseases
Disease.Cause the reduction of operating efficiency because body is ailing, increase insomniac's psychological pressure, such vicious circle, nerve occurs
Weak, endocrinopathy etc., or even it is developed to mental disease to more serious direction.
So the pharmaceutical requirements for treating insomnia increase, the medicine for treating insomnia is also invented and produced with various states.So
And not only effect cannot be affirmed these medicines mostly, also, hypnotic be mostly in vivo by liver, renal metabolism,
Long-term use can increase the burden of liver kidney, and some can also cause liver enlargement, hepatalgia, jaundice, edema, albuminuria, blood urine
And the liver and kidney dysfunction such as nausea, abdominal distension, poor appetite, constipation and stomach reaction.Some hypnotics also result in lassitude,
The accumulation of poisoning symptom such as hypophrenia, drop in blood pressure, or even cause respiratory and circulatory function obstacle situation.Therefore, the elderly applies
Stable class medicine more should be careful.
Therefore it provides Small side effects, and improve sleeper effect and significantly have great importance.
The content of the invention
In view of this, the technical problem to be solved in the present invention is to provide a kind of composition for improving sleep and its preparation side
Method, the significant effect provided by the invention for improving the composition slept and having no side effect and improving sleep.
The invention provides a kind of composition for improving sleep, including apple stem cell freeze-dried powder, epiphysin and vitamin
B6。
Wherein, apple stem cell not only can quickly activate the stem cell of dormancy in body, but also can active remediation by
The hyperplasia of the stem cell of damage, stimulating human horn cell and fibroblast, increase skin elasticity, there is anti-aging and beauty to give birth to
The special effect of hair.Apple stem cell has the characteristic of plant stem cell：Powerful anti-oxidant and activity of fighting against senium, can significantly improve people
Body immunity and anti-cancer ability.
Epiphysin is a kind of endogenous material, and secretion has circadian rhythm, is typically peaked in morning 2-3 points.Night
Between the height of melatonin levels directly influence the quality of sleep.With advancing age, the epiphysin of itself secretion is bright in vivo
It is aobvious to decline, reduce 10-15% within average every 10 years, cause sleep disordered and a series of functional disturbances, and melatonin levels reduce,
It is one of important symbol of mankind's brain aging that sleep, which is reduced,.
Vitamin B6Act predominantly on the blood of human body, muscle, nerve, skin etc..Major function has the synthesis of antibody, disappeared
The manufacturing of hydrochloric acid in gastric juice in change system, fat and protein utilization, maintain sodium/potassium balance (stable nervous system) and improve to sleep.
Be deficient in vitamin B6 logical disease, and the symptoms such as low poor appetite, food utilization, vomiting, insomnia are had when general lack of.It is serious to lack
Weary person have anaemia, arthritis, child's spasm, headache, etc..
The present invention can have after being compounded apple stem cell freeze-dried powder, epiphysin and vitamin B6 to be significantly improved
The function of sleep, and have no toxic side effect.Experiment shows that the significant effect of the improvement sleep of composition provided by the invention is better than single
Solely use each component effect, through acute toxicity test in mice result show mouse do not note abnormalities, do not occur poisoning or
The phenomena of mortality.And composition provided by the invention can effectively improve yellow Jackets sub-threshold dose under 0.5g/kg dosage
Mouse sleep incidence.
In an embodiment of the present invention, the mass ratio of apple stem cell freeze-dried powder, epiphysin and vitamin B6 for (20~
60)：(0.5~1)：(0.5~1).
In certain embodiments, the mass ratio of apple stem cell freeze-dried powder, epiphysin and vitamin B6 is 20:0.5:0.5.
In certain embodiments, the mass ratio of apple stem cell freeze-dried powder, epiphysin and vitamin B6 is 20:0.5:1.
In certain embodiments, the mass ratio of apple stem cell freeze-dried powder, epiphysin and vitamin B6 is 20:1:0.5.
In certain embodiments, the mass ratio of apple stem cell freeze-dried powder, epiphysin and vitamin B6 is 20:1:1.
In certain embodiments, the mass ratio of apple stem cell freeze-dried powder, epiphysin and vitamin B6 is 60:0.5:0.5.
In certain embodiments, the mass ratio of apple stem cell freeze-dried powder, epiphysin and vitamin B6 is 60:0.5:1.
In certain embodiments, the mass ratio of apple stem cell freeze-dried powder, epiphysin and vitamin B6 is 60:1:0.5.
In certain embodiments, the mass ratio of apple stem cell freeze-dried powder, epiphysin and vitamin B6 is 60:1:1.
In certain embodiments, the mass ratio of apple stem cell freeze-dried powder, epiphysin and vitamin B6 is 30:0.7:0.7.
The apple stem cell freeze-dried powder that the present invention uses can be bought for market also to be prepared using method provided by the invention,
It is implemented all within protection scope of the present invention.Preferably, apple stem cell freeze-dried powder is with method system provided by the invention
, this method using apple new life branch as raw material, after induced synthesis callus Multiplying culture and expand culture obtain apple
Stem cell, then be made after freezing.The preparation that the present invention provides composition, the freeze-dried powder are directly used in after the apple stem cell is lyophilized
Middle flavones and polyphenol content enrich.
The preparation method of apple stem cell freeze-dried powder comprises the following steps：
Step 1：The sterilization of apple new life branch, after removing xylem and marrow, inducing culture is inoculated in, Fiber differentiation obtains
Cambial cell；
Step 2：The cambial cell accesses proliferated culture medium, shaking table culture obtains unicellular through squamous subculture；
Step 3：The unicellular proliferated culture medium that is inoculated in is cultivated through expanding, obtains apple stem cell；
Step 4：It will freeze, crush after the apple stem cell multigelation, obtain apple stem cell freeze-dried powder；
The inducing culture is the MS solid mediums containing NAA, BA and caseinhydrolysate；
The proliferated culture medium is the MS fluid nutrient mediums containing 2,4-D, BA, caseinhydrolysate and activated carbon.
In an embodiment of the present invention, sterilization comprises the following steps：
Step 1：After apple new life branch is rinsed with water, the L-AA solution using concentration as 10~1000mg/L soaks
Bubble；
Step 2：With aseptic water washing after using volume fraction as 60%~95% ethanol water immersion；
Step 3：With aseptic water washing after using volume fraction as 10%~100% aqueous hydrogen peroxide solution immersion.
Preferably, the length of apple new life branch is 10cm.
Preferably, the concentration of L-AA is 120mg/L.
Preferably, the time soaked described in step 1 is 1min.
Preferably, the volume fraction of ethanol is 75% in ethanol water.
Preferably, the time soaked described in step 2 is 1min.
Preferably, the volume of sterilized water is 2L in step 2.
Preferably, the volume fraction of hydrogen peroxide is 30% in aqueous hydrogen peroxide solution.
Preferably, the time soaked described in step 3 is 20min.
Preferably, the time rinsed described in step 3 is 10min.
Carrying out sterilization to apple new life branch with method for disinfection provided by the invention can sufficiently kill except apple innovation
Entrained fungi, bacterium or virus on bar.To ensure that apple new life branch is not disturbed in incubation, so as to ensure
Cell on apple new life branch more quickly dedifferentes.And test and show, method provided by the invention can be effectively by apple
Fruit new life branch cell de-differentiation, through inducing and expanding culture, substantial amounts of apple stem cell can be obtained within 48 days or so.
In embodiments of the present invention, the mass ratio of NAA, BA and caseinhydrolysate is 0.5：2：1.
Preferably, NAA concentration is 0.5mg/L in inducing culture.
Preferably, BA concentration is 2.0mg/L in inducing culture.
Preferably, the concentration of caseinhydrolysate is 1.0mg/L in inducing culture.
Preferably, inducing culture is solid medium, wherein the mass fraction of agar is 1%.
Preferably, the pH value of inducing culture is 6.0.
In an embodiment of the present invention, the inoculation described in step 1 is that the apple innovation of xylem and marrow is removed per g
Bar is inoculated in 10cm2Inducing culture.
In certain embodiments, Fiber differentiation is light culture；Temperature is 25 DEG C；Time is 10 days.
After Fiber differentiation, Cambium tissue is the tabular tissue of homogeneous growth, and its hetero-organization is then to be irregular
Assemble growth-gen.
In certain embodiments, squamous subculture is light culture；Temperature is 25 DEG C；Time is 10 days.
In an embodiment of the present invention, squamous subculture is inoculated in 10cm using culture medium is run into per g cambial cells2's
Inducing culture.
Through the squamous subculture of 10 days, cambial cell formed callus, per 10cm2Inducing culture on there are about callus
Organize 20.8g.
In some embodiments, in proliferated culture medium, 2,4-D, BA, the mass ratio of caseinhydrolysate and activated carbon be 2:1:1:
1。
Preferably, 2,4-D concentration is 2.0mg/L in proliferated culture medium.
Preferably, BA concentration is 1.0mg/L in proliferated culture medium.
Preferably, the concentration of caseinhydrolysate is 1.0mg/L in proliferated culture medium.
Preferably, the concentration of activated carbon is 1.0mg/L in proliferated culture medium.
Preferably, the pH value of proliferated culture medium is 6.0.
In an embodiment of the present invention, per g callus access 20mL proliferated culture mediums.
Preferably, sterilized a diameter of 0.5cm bead is also added during shaking table culture, in proliferated culture medium.
Preferably, the addition of bead is 100g/L proliferated culture mediums.
In embodiment, culture uses triangular flask, and 200mL proliferated culture mediums are added in 1000mL triangular flasks.
In an embodiment of the present invention, the condition of shaking table culture is per hour with 20W incandescent lamp irradiation 10min.
In certain embodiments, the temperature of shaking table culture is 25 DEG C；Rotating speed is 120 revs/min；Time is 14 days.
Preferably, fresh proliferated culture medium is added after 7 days in shaking table culture.
Preferably, the volume for adding fresh proliferated culture medium is 2 times of former proliferated culture medium.
After adding fresh proliferated culture medium, wherein the density of cell is 1 × 105/ml。
Shaking table culture is carried out using method provided by the invention, callus turns into single cell suspension, cell proliferation rate
Comparatively fast, filtered with the stainless steel mesh of 100 mesh, removing cell mass, bead and residue, acquisition cell density about 0.5~1 ×
106/ml。
Preferably, the density being inoculated with step 3 is 1 × 104/ml。
In certain embodiments, expanding culture is specially：5L proliferated culture mediums, training are added in 20L bioreactor
It is 1 × 10 that proliferated culture medium to cell concentration is added after supporting 7 days5/ ml, it is further cultured for 7 days.
The preparation method of improvement sleep composition provided by the invention is, by apple stem cell freeze-dried powder, epiphysin and dimension
Raw plain B6 mixing, being made improves sleep composition.
Specifically, the preparation method of improvement sleep composition provided by the invention is：By apple stem cell freeze-dried powder at 4 DEG C
Crushed in low temperature, cross 100 mesh sieves；Epiphysin and vitamin B6 are crushed respectively, cross 100 mesh sieves after with the apple after sieving
Stem cell freeze-dried powder mixes.
Present invention also offers a kind of health products for improving sleep, including the composition provided by the invention for improving sleep.
Preferably, the health products provided by the invention for improving sleep also include acceptable auxiliary material in health products.
Preferably, the formulation of radiation-resistant health products provided by the invention is tablet.
The invention provides a kind of tablet for improving sleep, including the composition provided by the invention for improving sleep, disintegration
Agent, diluent, adhesive, thickener and lubricant.
In certain embodiments, the tablet provided by the invention for improving sleep includes the group provided by the invention for improving sleep
Compound, microcrystalline cellulose, pregelatinized starch, cross-linked carboxymethyl cellulose are received, PVP K30, silica and magnesium stearate.
In certain embodiments, in the tablet provided by the invention for improving sleep, the group provided by the invention for improving sleep
Compound, microcrystalline cellulose, pregelatinized starch, cross-linked carboxymethyl cellulose are received, PVP K30, silica and magnesium stearate
Mass ratio is (21~62)：(20~40)：(10~20)：(3~6)：(2~3)：0.5：1.
Preferably, in the tablet provided by the invention for improving sleep, the composition, micro- provided by the invention for improving sleep
Crystalline cellulose, pregelatinized starch, cross-linked carboxymethyl cellulose are received, the mass ratio of PVP K30, silica and magnesium stearate is
31.4：30：15：4：2.5：0.5：1.
The preparation method of the tablet provided by the invention for improving sleep is the combination that the improvement for being supplied to the present invention is slept
After thing, disintegrant and diluent mixing, add the aqueous solution softwood of adhesive, through pelletizing, dry after with thickener, lubricant
The tablet for improving sleep is made in mixing, tabletting.
Specifically, this method specifically includes：
Step 1：PVP K30 is configured to the aqueous solution that mass fraction is 5%~10%, as adhesive；
Step 2：4 DEG C, by the composition provided by the invention for improving sleep, microcrystalline cellulose, pregelatinized starch, crosslinking carboxylic
Sodium carboxymethylcellulose pyce mixes, and is pelletized after adding described adhesive softwood；
Step 3：The obtained improvement of tabletting after mixing with silica, magnesium stearate after whole grain is dried after placing 12 hours to sleep
The tablet of dormancy.
The present invention can have after being compounded apple stem cell freeze-dried powder, epiphysin and vitamin B6 to be significantly improved
The function of sleep, and have no toxic side effect.Experiment shows that the significant effect of the improvement sleep of composition provided by the invention is better than single
Solely use each component effect, through acute toxicity test in mice result show mouse do not note abnormalities, do not occur poisoning or
The phenomena of mortality.And composition provided by the invention can effectively improve yellow Jackets sub-threshold dose under 0.5g/kg dosage
Mouse sleep incidence.
Embodiment
The invention provides a kind of composition for improving sleep and preparation method thereof, those skilled in the art can use for reference this
Literary content, it is suitably modified technological parameter realization.In particular, all similar replacements and change are to art technology
It is it will be apparent that they are considered as being included in the present invention for personnel.The method of the present invention and application are by preferable
Embodiment is described, related personnel substantially can not depart from present invention, in spirit and scope to methods herein and
Using being modified or suitably change with combining, to realize and using the technology of the present invention.
The instrument that the present invention uses is all common commercially available product, can all be bought in market.
With reference to embodiment, the present invention is expanded on further：
The preparation of the apple stem cell freeze-dried powder of embodiment 1
1st, the formula of culture medium：
1.1 inducing culture：MS solid medium+NAA0.5mg/L+BA 2mg/L+CH 1mg/L, pH value 6.0；
1.2 proliferated culture medium：MS fluid nutrient medium+2,4-D 2.0mg/L+BA 1.0mg/L+CH1.0mg/L+AC
1.0mg/L, pH value 6.0；
2nd, the sterilization of apple branch
The apple tree without fertilizer and pesticide pollution, green planting is selected, 10 10cm length are selected according to certain judgment criteria
Newborn branch；Newborn branch surface attachments are washed with sterilized water 2L；Newborn branch is put into 120mg/L L-AAs
Immersion 1 minute；Newborn branch is put into 75% ethanol solution and sterilized 1 minute, it is clean with 2L aseptic water washings；Place into
Sterilized 20 minutes in 30% hydrogenperoxide steam generator, and with aseptic water washing 10 minutes.
3rd, the induction of Cambium tissue
The apple new life branch of sterilizing is peeled off into the tissue such as forming layer, bast, cortex and epidermis with vaccinating lancet, abandons wood
Matter portion and marrow；The tissue of acquisition is seeded in fresh solid medium, inoculation 10cm is organized with every 1g2Solid medium
Density is inoculated with, and is continuously cultivated 10 days for 25 DEG C in controlled darkroom, is obtained Cambium tissue.
4th, squamous subculture Cambium tissue
The Cambium tissue of regeneration and its hetero-organization are separated；Inoculation 10cm is organized with every 1g2Solid medium it is close
Degree, the Cambium tissue of regeneration is seeded in fresh solid medium, is continuously cultivated 10 days for 25 DEG C in controlled darkroom,
Obtain callus.
5th, unicellular culture
200ml fluid nutrient mediums, the 20g of sterilization treatment, diameter 0.5cm bead are added in 1000ml triangular flasks；
10g callus is transferred in triangular flask；Triangular flask is placed in 25 DEG C, cultivated on 120 revs/min of shaking table, per small
When irradiated 10 minutes with 20W incandescent lamp；7th day, it was wherein cell to add the fresh fluid nutrient mediums of 400ml to cell density
Density be 1 × 105/ ml, then proceed to culture 7 days；Filtered after culture with the stainless steel mesh of 100 mesh, remove cell mass, glass
Glass pearl and residue, obtain single cell suspension,
6th, amplification culture
Single cell suspension is transferred in 20L bioreactors and carries out large-scale culture, accommodates 5L's in bioreactor
Proliferated culture medium.When cultivating the 7th day, 10L fresh liquid culture medium is added, cell suspension is obtained after continuing culture.
7th, the preparation of freeze-dried powder
Apple stem cell suspension is filtered with 1um filters, removes culture medium, and the viscose shape cell for harvesting retention takes out；Will
Cell is put into pre-freeze 3 hours in -30 DEG C of refrigerator；It is put into super low temperature quick frozen 2 hours in liquid nitrogen container；Take out after thawing 30 minutes,
Place into it is quick-frozen in pre-freeze and liquid nitrogen container in -30 DEG C of refrigerators, 2 times successively；Cell is taken out, maintains 10 minutes, enters low at room temperature
In warm frozen vacuum dryer, after vacuum freeze drying, 60 mesh freeze-dried powders are ground at a high speed.
Embodiment 2 improves the preparation of sleep composition
First by apple stem cell freeze-dried powder 100g, epiphysin 0.5g, vitamin B6 2.5g, it is (wherein, right to be crushed respectively
Apple stem cell freeze-dried powder liquid nitrogen grinding, epiphysin and vitamin B6 carry out common grinding), 100 mesh sieves are crossed, then by powder
Broken, screened material is well mixed in mixer.
Embodiment 3 improves the preparation of sleep composition
First by apple stem cell freeze-dried powder 100g, epiphysin 2.5g, vitamin B6 5g, crushed respectively (wherein, to apple
Dried fruit cell freeze-dried powder liquid nitrogen grinding, epiphysin and vitamin B6 carry out common grinding), 100 mesh sieves are crossed, then will crush,
Screened material is well mixed in mixer.
Implementing 4 improves the preparation of sleep composition
First by apple stem cell freeze-dried powder 100g, epiphysin 5g, vitamin B6 2.5g, crushed respectively (wherein, to apple
Dried fruit cell freeze-dried powder liquid nitrogen grinding, epiphysin and vitamin B6 carry out common grinding), 100 mesh sieves are crossed, then will crush,
Screened material is well mixed in mixer.
Embodiment 5 improves the preparation of sleep composition
First by apple stem cell freeze-dried powder 100g, epiphysin 5g, vitamin B6 5g, crushed respectively (wherein, to apple
Stem cell freeze-dried powder liquid nitrogen grinding, epiphysin and vitamin B6 carry out common grinding), 100 mesh sieves are crossed, then will have been crushed, mistake
The material of sieve is well mixed in mixer.
Embodiment 6 improves the preparation of sleep composition
First by apple stem cell freeze-dried powder 150g, epiphysin 1.25g, vitamin B6 1.25g, crushed respectively (wherein,
To apple stem cell freeze-dried powder liquid nitrogen grinding, epiphysin and vitamin B6 carry out common grinding), 100 mesh sieves are crossed, then by powder
Broken, screened material is well mixed in mixer.
Embodiment 7 improves the preparation of sleep composition
First by apple stem cell freeze-dried powder 150g, epiphysin 1.25g, vitamin B6 2.5g, crushed respectively (wherein,
To apple stem cell freeze-dried powder liquid nitrogen grinding, epiphysin and vitamin B6 carry out common grinding), 100 mesh sieves are crossed, then by powder
Broken, screened material is well mixed in mixer.
Embodiment 8 improves the preparation of sleep composition
First by apple stem cell freeze-dried powder 150g, epiphysin 2.5g, vitamin B6 1.25g, crushed respectively (wherein,
To apple stem cell freeze-dried powder liquid nitrogen grinding, epiphysin and vitamin B6 carry out common grinding), 100 mesh sieves are crossed, then by powder
Broken, screened material is well mixed in mixer.
Embodiment 9 improves the preparation of sleep composition
First by apple stem cell freeze-dried powder 150g, epiphysin 2.5g, vitamin B6 2.5g, it is (wherein, right to be crushed respectively
Apple stem cell freeze-dried powder liquid nitrogen grinding, epiphysin and vitamin B6 carry out common grinding), 100 mesh sieves are crossed, then by powder
Broken, screened material is well mixed in mixer.
Embodiment 10 improves the preparation of sleep composition
First by apple stem cell freeze-dried powder 150g, epiphysin 3.5g, vitamin B6 3.5g, it is (wherein, right to be crushed respectively
Apple stem cell freeze-dried powder liquid nitrogen grinding, epiphysin and vitamin B6 carry out common grinding), 100 mesh sieves are crossed, then by powder
Broken, screened material is well mixed in mixer.
Embodiment 11 improves the preparation of sleeping tablets
10.3g PVP K30s are configured to the aqueous solution that mass fraction is 7% with purified water, as adhesive；4 DEG C of bars
Under part, 103g hands over composition, 98g microcrystalline celluloses, 49g pregelatinized starch, the 14.7g of the improvement sleep that embodiment 2 provides
Join sodium carboxymethylcellulose mixing, pelletized after adding described adhesive softwood；Done after placing 12 hours in drying box
Dry, dried particle carries out whole grain using fast finishing agent, is mixed after whole grain with 2.45g silica, 4.9g magnesium stearates
After conjunction, the obtained tablet for improving sleep of tabletting is carried out using tablet press machine.
Embodiment 12 improves the preparation of sleeping tablets
9g PVP K30s are configured to the aqueous solution that mass fraction is 8% with purified water, as adhesive；Under the conditions of 4 DEG C
Composition, 90g microcrystalline celluloses, the 45g pregelatinized starch 14g of the improvement sleep that embodiment 3 provides are crosslinked carboxylic first by 107.5g
Base sodium cellulosate mixes, and is pelletized after adding described adhesive softwood；It is dried, dries in drying box after placing 12 hours
Particle afterwards carries out whole grain using fast finishing agent, after being mixed after whole grain with 2g silica, 4.1g magnesium stearates, using pressure
Piece machine carries out tabletting and the tablet for improving sleep is made.
Embodiment 13 improves the preparation of sleeping tablets
8g PVP K30s are configured to the aqueous solution that mass fraction is 9% with purified water, as adhesive；4 DEG C of conditions
Under, 107.5g hands over composition, 83g microcrystalline celluloses, 41.5g pregelatinized starch, the 13g of the improvement sleep that embodiment 4 provides
Join sodium carboxymethylcellulose mixing, pelletized after adding described adhesive softwood；Done after placing 12 hours in drying box
Dry, dried particle carries out whole grain using fast finishing agent, is mixed after whole grain with 1.7g silica, 6g magnesium stearates
Afterwards, the obtained tablet for improving sleep of tabletting is carried out using tablet press machine.
Embodiment 14 improves the preparation of sleeping tablets
7.3g PVP K30s are configured to the aqueous solution that mass fraction is 10% with purified water, as adhesive；4 DEG C of bars
Under part, 110g hands over composition, 79g microcrystalline celluloses, 39g pregelatinized starch, the 12.8g of the improvement sleep that embodiment 5 provides
Join sodium carboxymethylcellulose mixing, pelletized after adding described adhesive softwood；Done after placing 12 hours in drying box
Dry, dried particle carries out whole grain using fast finishing agent, is mixed after whole grain with 1.5g silica, 3.72g magnesium stearates
After conjunction, the obtained tablet for improving sleep of tabletting is carried out using tablet press machine.
Embodiment 15 improves the preparation of sleeping tablets
9.7g PVP K30s are configured to the aqueous solution that mass fraction is 9% with purified water, as adhesive；4 DEG C of conditions
Under, composition, 104g microcrystalline celluloses, 52g pregelatinized starch, the 17g of the improvement sleep that embodiment 6 provides are crosslinked by 152.5g
Sodium carboxymethylcellulose mixes, and is pelletized after adding described adhesive softwood；It is dried after placing 12 hours in drying box,
Dried particle carries out whole grain using fast finishing agent, is mixed after whole grain with 1.8g silica, 3.72g magnesium stearates
Afterwards, the obtained tablet for improving sleep of tabletting is carried out using tablet press machine.
Embodiment 16 improves the preparation of sleeping tablets
8.9g PVP K30s are configured to the aqueous solution that mass fraction is 10% with purified water, as adhesive；4 DEG C of bars
Under part, 153.75g by embodiment 7 provide improvement sleep composition, 132g microcrystalline celluloses, 53g pregelatinized starch,
16.5g Ac-Di-Sols mix, and are pelletized after adding described adhesive softwood；After placing 12 hours in drying box
Be dried, dried particle using fast finishing agent carry out whole grain, after whole grain with 1.6g silica, 3.3g stearic acid
After magnesium mixing, the obtained tablet for improving sleep of tabletting is carried out using tablet press machine.
Embodiment 17 improves the preparation of sleeping tablets
8.7g PVP K30s are configured to the aqueous solution that mass fraction is 10% with purified water, as adhesive；4 DEG C of bars
Under part, composition, 96g microcrystalline celluloses, 54g pregelatinized starch, the 16.5g of the improvement sleep that 153.75g provides embodiment 8
Ac-Di-Sol mixes, and is pelletized after adding described adhesive softwood；Carried out after placing 12 hours in drying box
Dry, dried particle carries out whole grain using fast finishing agent, is mixed after whole grain with 1.5g silica 3g magnesium stearates
Afterwards, the obtained tablet for improving sleep of tabletting is carried out using tablet press machine.
Embodiment 18 improves the preparation of sleeping tablets
8.3g PVP K30s are configured to the aqueous solution that mass fraction is 10% with purified water, as adhesive；4 DEG C of bars
Under part, composition, 94.2g microcrystalline celluloses, 55g pregelatinized starch, the 16.6g of the improvement sleep that 155g provides embodiment 9
Ac-Di-Sol mixes, and is pelletized after adding described adhesive softwood；Carried out after placing 12 hours in drying box
Dry, dried particle carries out whole grain using fast finishing agent, is mixed after whole grain with 1.4g silica, 2.8g magnesium stearates
After conjunction, the obtained tablet for improving sleep of tabletting is carried out using tablet press machine.
Embodiment 19 improves the preparation of sleeping tablets
7.5g PVP K30s are configured to the aqueous solution that mass fraction is 10% with purified water, as adhesive；4 DEG C of bars
Under part, 157g hands over composition, 150g microcrystalline celluloses, 75g pregelatinized starch, the 20g of the improvement sleep that embodiment 10 provides
Join sodium carboxymethylcellulose mixing, pelletized after adding described adhesive softwood；Done after placing 12 hours in drying box
Dry, dried particle carries out whole grain using fast finishing agent, is mixed after whole grain with 2.5g silica, 5g magnesium stearates
Afterwards, the obtained tablet for improving sleep of tabletting is carried out using tablet press machine.
The preparation of the reference substance 1 of comparative example 1
7.5g PVP K30s are configured to the aqueous solution that mass fraction is 10% with purified water, as adhesive；4 DEG C of bars
Under part, 157g apple stem cells freeze-dried powder, 150g microcrystalline celluloses, 75g pregelatinized starch, 20g Ac-Di-Sols
Mixing, pelletized after adding described adhesive softwood；It is dried after placing 12 hours in drying box, dried particle is adopted
Whole grain is carried out with fast finishing agent, after being mixed after whole grain with 2.5g silica, 5g magnesium stearates, is pressed using tablet press machine
The tablet for improving sleep is made in piece.
The preparation of the reference substance 2 of comparative example 2
7.5g PVP K30s are configured to the aqueous solution that mass fraction is 10% with purified water, as adhesive；4 DEG C of bars
Under part, 157g epiphysins, 150g microcrystalline celluloses, 75g pregelatinized starch, the mixing of 20g Ac-Di-Sols, institute is added
Pelletized after stating adhesive softwood；It is dried after placing 12 hours in drying box, dried particle uses fast finishing
Agent carries out whole grain, after being mixed after whole grain with 2.5g silica, 5g magnesium stearates, carries out tabletting using tablet press machine and improvement is made
The tablet of sleep.
The preparation of the reference substance 3 of comparative example 3
7.5g PVP K30s are configured to the aqueous solution that mass fraction is 10% with purified water, as adhesive；4 DEG C of bars
Under part, 157g vitamin B6s, 150g microcrystalline celluloses, 75g pregelatinized starch, the mixing of 20g Ac-Di-Sols, add
Pelletized after described adhesive softwood；It is dried after placing 12 hours in drying box, dried particle is using quick whole
Manage agent and carry out whole grain, after being mixed after whole grain with 2.5g silica, 5g magnesium stearates, be made and changed using tablet press machine progress tabletting
The tablet of kind sleep.
Embodiment 20 improves the acute toxicity test in mice of sleeping tablets
Test reference《Health food is examined and assessment technique specification》(2003 editions).
The mouse for taking body weight to be 18-22g, sets 5 groups altogether, every group 10, male and female half and half, respectively according to 4.64g/kg, 10g/
Kg, 21.5g/kg, 46.4g/kg gavage give the tablet of the offer of the embodiment of the present invention 19, separately set blank control group, and control group fills
Stomach distilled water, according to mouse 20ml/kgBW amount, the gastric infusion in 12 hours, fasting 15 hours, can't help before mouse administration
Water, end a hunger strike within 3 hours after administration.Continuous Observation 14 days after administration stops, observing daily mouse spirit, appetite, drinking-water, activity and in
Malicious situation, and record intoxication conditions, death time and the mortality of mouse.As a result such as table 1：
The acute toxicity test in mice result of table 1
As a result show：Dosage according to 4.64g/kg, 10g/kg, 21.5g/kg and 46.4g/kg carries out gavage to mouse and given
Medicine is tested, and is observed 14 days, and whole mouse spirit, activity, mouth, eye nose have no secretion, spirit, appetite, situations such as drinking water and right
It is more without exception according to organizing.More than known to after 46.4g/kg, be not poisoned or dead mouse situation.According to toxicological evaluation mark
Standard, can be unlike measure median lethal dose (LD50), i.e. LD50>15g/kg.As a result show, above-mentioned products obtained therefrom belongs to non-toxic type guarantor
Strong product.
Tablet test result made from other embodiments of the invention is similarly.
Embodiment 21 improves influence of the sleeping tablets to mouse weight
Take the mouse that body weight is 18-22g, each 40 of male and female, respectively according to 1.0g/kg, 3.0g/kg, 5.0g/kg dosage
Gavage gives the tablet of the offer of the embodiment of the present invention 19, separately sets blank control group, control group gavage distilled water, according to mouse
20ml/kgBW amount, the gastric infusion in 12 hours, fasting 15 hours, can't help water, ends a hunger strike within 3 hours after administration before mouse administration.
Successive administration 30 days, during which observe the changes of weight of mouse.As a result such as table 2：
Test mice body weight increase situation during table 2 is tested
As shown in Table 2, body weight increase does not have significant difference between experiment mice each group dosage, and products obtained therefrom is small to male and female
Without influence, can obtain above-mentioned products obtained therefrom does not influence mouse body weight on tested material body weight.Observe and also found, the above-mentioned products obtained therefrom of gavage
Mouse compared with control group, hair and skin are more smooth, activity it is more flexible.
To tablet test result made from other embodiments of the invention similarly.
Embodiment 22 improves sleeping tablets and extends the experiment of the yellow Jackets length of one's sleep
Each 80 of male and female mouse, is randomly divided into 4 groups, gives tablet and comparative example that isodose embodiment 19 provides respectively
1~3 reference substance provided.
Wherein, experimental group 1~3 gives the water of equivalent with dosage 0.1g/kg, 0.5g/kg, 1.5g/kg gavage, 0 dosage group.
Gavage 1 time daily, continuous gavage 15 days.After last gavage 15min, 50mg/kgBW amobarbitals are injected intraperitoneally to each group animal
Sodium, injection volume 0.3mL/20gBW, to right transmitting hour as sleep criterion, whether observation given the test agent extends penta bar
Than the appropriate sodium length of one's sleep, mouse hypnagogic latency time such as table 3：
The record sheet of the mouse hypnagogic latency time of table 3
As a result：From table 3, compared with 0 dosage group, giving health products length of one's sleep difference provided by the invention has system
Meter learns meaning (P<0.05) length of one's sleep for yellow Jackets induction that, to be embodiment 19 can extend in 0.5g/kg dosage group.
Observation also found that for the mouse of the above-mentioned products obtained therefrom of gavage compared with 0 dosage group, hair and skin are more smooth, and activity is cleverer
It is living.To tablet test result made from other embodiments of the invention similarly.And the testing result of control sample 1~3 is shown,
Compared with 0 dosage group, sleep mouse incidence does not have statistical significance (P>0.05) it is, that the dosage of control sample 1~3 exists
1.5g/kg can not effectively improve yellow Jackets sub-threshold dose mouse sleep incidence.
Embodiment 23 improves the experiment of sleeping tablets yellow Jackets sub-threshold dose hypnotic dose
Preliminary experiment is first passed through to determine pentobarbital sodium sub-threshold lull dosage, be 85%~90% mouse righting reflex not
The yellow Jackets maximum sub-threshold dose of disappearance is 15mg/kg.
Each 80 of male and female mouse, is randomly divided into 4 groups, gives tablet and comparative example that isodose embodiment 19 provides respectively
1~3 reference substance provided.
Wherein, experimental group 1~3 gives the water of equivalent with dosage 0.1g/kg, 0.5g/kg, 1.5g/kg gavage, 0 dosage group.
Gavage 1 time daily, continuous gavage 15 days.Occur 12min before peak value after last gavage, give each group animal intraperitoneal injection 15mg/kg
The yellow Jackets of dosage, transmitting is righted using mouse and disappeared up to more than 2min persons as sleep standard, records sleep animal in 20min
Number, continuous record one month, observes whether above-mentioned products obtained therefrom can improve yellow Jackets sub-threshold dose sleep mouse incidence.
Table 4 records mouse sleep record sheet
Note：* the P compared with 0 dosage group is shown<0.05, * * shows the P compared with 0 dosage group<0.01.
# shows the P compared with 0 dosage group>0.05, ## shows the P compared with 0 dosage group>0.05.
For each dosage group compared with 0 dosage group, sleep mouse incidence has statistical significance (P<0.05), it is embodiment
The dosage of tablet made from 19 can improve yellow Jackets sub-threshold dose mouse sleep incidence in 0.5g/kg.To the present invention its
Tablet test result made from his embodiment is similarly.
And the testing result for providing comparative example 1~3 reference substance shows that each dosage group is compared with 0 dosage group, sleep mouse
Incidence does not have statistical significance (P>0.05) it is, that control sample 1~3 can not effectively improve amobarbital in 1.0g/kg
Sodium sub-threshold dose mouse sleep incidence.
It the above is only the preferred embodiment of the present invention, it is noted that come for those skilled in the art
Say, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should be regarded as
Protection scope of the present invention.
Claims (9)
1. a kind of composition for improving sleep, it is characterised in that by apple stem cell freeze-dried powder, epiphysin and vitamin B6 group
Into；
The mass ratio of the apple stem cell freeze-dried powder, epiphysin and vitamin B6 is (20~60)：(0.5~1)：(0.5~
1)。
2. composition according to claim 1, it is characterised in that apple stem cell freeze-dried powder, epiphysin and the Wei Sheng
Plain B6 mass ratio is 30:0.7:0.7.
3. composition according to claim 1, it is characterised in that the preparation method of wherein apple stem cell freeze-dried powder includes
Following steps：
Step 1：The sterilization of apple new life branch, after removing xylem and marrow, inducing culture is inoculated in, Fiber differentiation is formed
Confluent monolayer cells；
Step 2：The cambial cell accesses proliferated culture medium, shaking table culture obtains unicellular through squamous subculture；
Step 3：The unicellular proliferated culture medium that is inoculated in is cultivated through expanding, obtains apple stem cell；
Step 4：It will freeze, crush after the apple stem cell multigelation, obtain apple stem cell freeze-dried powder；
The inducing culture is the MS solid mediums containing NAA, BA and caseinhydrolysate；
The proliferated culture medium is the MS fluid nutrient mediums containing 2,4-D, BA, caseinhydrolysate and activated carbon.
4. the preparation method of the improvement sleep composition as described in any one of claims 1 to 3, it is characterised in that by dried apple slices
Cell freeze-dried powder, epiphysin and microorganism B6 mixing, being made improves sleep composition.
5. a kind of health products for improving sleep, it is characterised in that including the composition described in any one of claims 1 to 3.
6. a kind of tablet for improving sleep, it is characterised in that including the composition described in any one of claims 1 to 3, disintegration
Agent, diluent, adhesive, thickener and lubricant.
7. the tablet according to claim 6 for improving sleep, it is characterised in that including described in any one of claims 1 to 3
Composition, microcrystalline cellulose, pregelatinized starch, cross-linked carboxymethyl cellulose receive, PVP K30, silica and stearic acid
Magnesium.
8. the tablet according to claim 7 for improving sleep, it is characterised in that described in any one of claims 1 to 3
Composition, microcrystalline cellulose, pregelatinized starch, cross-linked carboxymethyl cellulose receive, PVP K30, silica and stearic acid
The mass ratio of magnesium is (21~62)：(20~40)：(10~20)：(3~6)：(2~3)：0.5：1.
9. the preparation method of the tablet of the improvement sleep as described in any one of claim 6~8, it is characterised in that will by right
After asking composition, disintegrant and diluent described in 1~3 any one to mix, the aqueous solution softwood of adhesive is added, through system
Grain, the tablet slept after drying with thickener, mix lubricant, the obtained improvement of tabletting.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102960718A (en) * | 2012-07-16 | 2013-03-13 | 赫康贸易(上海)有限公司 | Health products of plant stem cell |
| CN102973561A (en) * | 2012-11-23 | 2013-03-20 | 西安泰科迈医药科技有限公司 | Drug composite for improving sleep and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102960718A (en) * | 2012-07-16 | 2013-03-13 | 赫康贸易(上海)有限公司 | Health products of plant stem cell |
| CN102973561A (en) * | 2012-11-23 | 2013-03-20 | 西安泰科迈医药科技有限公司 | Drug composite for improving sleep and preparation method thereof |
Non-Patent Citations (2)
| Title |
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| 吃苹果改善睡眠;王学东;《中国中医药报》;20080321;第1页 * |
| 苹果体细胞悬浮培养及植株再生研究;李玉生等;《北方园艺》;20111231(第19期);第103-106页 * |
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