CN104856924B - A kind of preparation method of external application probiotic microcapsule - Google Patents
A kind of preparation method of external application probiotic microcapsule Download PDFInfo
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Abstract
The invention discloses a kind of preparation method of external application probiotic microcapsule, makes full use of plant extract and the synergistic effect of probiotics, one side concertedness improves survival rate and envelop rate, still further aspect, plant extraction liquid also has further effect to skin nursing.A kind of preparation method of external application probiotic microcapsule：(1), by Bifidobacterium, lactobacillus acidophilus, Lactobacillus rhamnosus lyophilized bacterium powder, be inoculated in activation process in triangular flask respectively, expand fermented and cultured, by the zymotic fluid of three kinds of bacterium according to volume ratio 3：3：4 are mixed, and obtain bacteria suspension；（2）, prepare nostoc extracting solution；（3）, by bacteria suspension, after nostoc extracting solution and sodium alginate soln add Tween 80 emulsification, cure, after wet capsule is formed, with filtered through gauze, then again with normal saline flushing 3 times.
Description
Technical field
The invention belongs to biomedicine technical field, and in particular to a kind of preparation method of external application probiotic microcapsule.
Background technology
In recent years, many bacterial fermentation products are applied to cosmetics, and among the application of external preparation for skin, such as two, which split yeast, carries
Take liquid, saccharomyces sake extracting solution etc..Probiotics cosmetics have attracted the change of whole industry.Its appearance make it is natural, healthy, have
The cosmetics concept of machine is more rooted in the hearts of the people.Probiotics cosmetics can be roughly divided into three classes：First, probiotic acid milk is made up
Product, the facial mask such as made using probiotic yogurt.Second, probiotics fermention liquid activity extract, third, directly being added in cosmetics
Enough probiotics and prebiotics.Three classes product respectively has each feature.First, probiotic yogurt is good cosmetics, it have than
The nutrition (a variety of amino acid and vitamin B complex) that milk is more more easy to be absorbed by organisms can be with nourishes rough skin, and lactic acid therein can
To keep skin to be in the acid range of health.Secondly, probiotics fermention liquid activity extract, as some active peptides obtain
It after purification, can comparatively fast promote the renewal with root of hair cell in skin, play the role of crease-resistant and hair care.Above all
The probiotics of activity suppresses the growth of skin surface spoilage organisms, protects the skin of sensitivity.Even some probiotics can stimulate endogenous
The synthesis of nerve acid amides, safeguards the normal lipid layer function of skin.Prebiotics is supplied to skin surface probiotic comestible in time,
Extend the probiotic action time.In Greece, active Yoghourt has a long history as a kind of natural sun-prevention nursing materials.In fact
There are many people to cook Yoghourt facial mask, if changing common sour milk facial mask into probiotic yogurt facial mask, moistening effect can be more preferable.Due to
Child skin is very delicate, the use of adult cosmetics is very inappropriate, may bring irremediable injury.Probioticsization
Cosmetic will be big promising in cosmetics for children field.
The most of probiotics raw materials of the prior art are all made as extracting solution form and add in cream or elite, mostly simply
Extracellular extract, although has skin many important physiological functions, but to obtain desired treatment, health-care effect, it is necessary to
There is probiotics living, and enough quantity must be reached, but probiotics is more fragile and easily by surrounding environment by itself
The influence of factor, causes probiotics survival period in the product developed short, after body is entered, and easily by skin secretion
Effect, the quantity for again resulting in probiotics soon decline, and the probiotics quantity for causing to eventually enter into skin seldom cannot
Effectively play its prebiotic effect.Probiotic microcapsule embedding techniques, makes probiotics after wrapping up layer by layer, can reach and resist and be resistant to
External environment is destroyed, and is preferably resolved the deficiency of probiotics itself, is effectively increased probiotics viable bacteria quantity.
The prior art has had the external application probiotic microcapsule embedded to lactobacillus.But its embedding rate and deposit
Motility rate is not ideal enough, and component is also relatively simple.
The present invention is intended to provide a kind of preparation method of external application probiotic microcapsule, makes full use of plant extract and probiotics
Synergistic effect, one side concertedness, which improves survival rate and envelop rate, still further aspect, plant extraction liquid, also has skin nursing
Further effect.
The content of the invention
The object of the present invention is to provide a kind of preparation method of external application probiotic microcapsule, plant extract and benefit are made full use of
The synergistic effect of raw bacterium, one side concertedness improve survival rate and envelop rate, still further aspect, and plant extraction liquid also protects skin
Reason has further effect.
In order to realize above-mentioned purpose, present invention employs following technical solution：
A kind of preparation method of external application probiotic microcapsule, its step are as follows：
(1), by Bifidobacterium, lactobacillus acidophilus, Lactobacillus rhamnosus lyophilized bacterium powder, be inoculated in respectively in triangular flask living
Change is handled, and expands fermented and cultured, the zymotic fluid of three kinds of bacterium is mixed, obtains bacteria suspension；
（2）, take fresh nostoc, crush, with microwave radiation, add boiling to boil 20-25min, filter while hot, in the dregs of a decoction again
Add water to continue to decoct, be repeated several times, merging filtrate, refined filtration, obtain nostoc extracting solution；
（3）, bacteria suspension, nostoc extracting solution and sodium alginate soln mixed, and it is 0.4% to spit to add concentration
Temperature 80 emulsifies, and is sprayed into emulsion as in the calcium chloride solution on magnetic stirring apparatus with sterile drive nozzle, after curing, treated wet
After capsule is formed, with filtered through gauze, normal saline flushing is then used again 3 times.
Preferably, the step（1）For：By Bifidobacterium, lactobacillus acidophilus, Lactobacillus rhamnosus lyophilized bacterium powder,
Activation process in triangular flask is inoculated in respectively, i.e., after 30~35 DEG C are cultivated 50~60h, expands fermented and cultured, when its zymotic fluid is lived
Bacteria concentration reaches 1010Cfu/g, terminates fermentation, by the zymotic fluid of three kinds of bacterium according to volume ratio 3：3：4 are mixed, and are obtained bacterium and are hanged
Liquid；
Preferably, the step（2）For：Fresh nostoc is taken, is crushed, with 3-5 points of microwave power 280-320W radiation
Clock, adds the boiling boiling 20-25min of 5 times of weight, filters while hot, continue to decoct to water is added in the dregs of a decoction, repeat extraction three times,
Merging filtrate, refined filtration, obtains nostoc extracting solution；
Preferably, the step（3）For：By bacteria suspension, nostoc extracting solution and sodium alginate soln are by volume
For 1：1：8 are mixed, and add concentration be 0.4% Tween 80 emulsify 18-20 minute after, with sterile drive nozzle will emulsification
Liquid is sprayed into as in the calcium chloride solution on magnetic stirring apparatus, after curing 10-12 minutes, after wet capsule is formed, with gauze mistake
Filter, then uses normal saline flushing 3 times again, and the sodium alginate soln concentration is 2%, and the concentration of calcium chloride solution is 2%, breast
Change and the immobilization time is 15 minutes.
Lactic acid bacteria is a kind of bacterium general name that energy metabolism produces lactic acid, and lactic acid bacteria is that the majority found so far is beneficial to human body
Harmless maximum micropopulation.And Bifidobacterium is the most beneficial lactobacillus of enteral, have antibacterial, it is anti-oxidant, antitumor,
Many-sided functions such as reducing blood lipid, enhancing be immune, and Bifidobacterium in enteron aisle and the service life of people have certain relation.With the age
Increase, inferior health or sick, the Bifidobacterium of enteral can substantially reduce, so the countermeasure that scholar and industry personnel take is raw
The food containing Bifidobacterium or medicine are produced for using, increases the quantity of Bifidobacterium in enteron aisle, to play the anti-ageing of Bifidobacterium
Always, the effect of diseases prevention.
Nostoc frond is in colloidal substance, spherical or other irregular shapes, blue-green or yellowish-brown.Cell is spherical, by more
Number cell is linked to be beads shape colony, outsourcing colloid.Heterocyst is located at the iuntercellular of filament, and young filament is frequently located in top.Breeding
When it is most produce akinetes, often to concatenate into chain.It is in grey black after dry in green during moistening.Attached submersed sand
Between stone or on dark and damp soil.It is distributed all over China, with the most famous of Sichuan production.It is edible.With nitrogen fixing capacity.Contain
There are a variety of amino acid and polysaccharide isoreactivity materials necessary to human body.With relieve inflammation or internal heat, improving eyesight, anti-aging, the therapeutic efficiency such as anti-infective.
Contain 18 kinds of amino acid in nostoc, wherein containing eight kinds of amino acid needed by human, dry matter total protein is up to 56% or so, thick fat
Fat 8.11%, carbohydrate 12.69%, ash content 10.88%, 30.98 mg/g of chlorophyll, 5.50 mg/g of ascorbic acid, vitamin
C content is higher than hawthorn more than 5 times close to fresh dates, 5 times higher than citrus；Vitamin B1, B2 are higher than general Homonemeae：Containing mineral matter 15
Kind, most abundant has phosphorus, sulphur, calcium, potassium, iron etc., and less has lead, silicon, magnesium, barium, germanium etc., and trace element has zinc, copper, manganese, also
There are starch and carbohydrate, its calcium content is higher than general vegetables, is a kind of fabulous Natural Rich Calcium nutraceutical.By celestial to wild Pueraria
The Analysis of Nutritive Composition of rice, draws Wild Nostoc protein content up to 48.61%, containing 17 kinds of amino acid, wherein containing 7 kinds of human body
Essential amino acid is 44.619%；Lysine, the content of threonine are higher especially in Wild Nostoc.But nostoc contains
Polysaccharide, sometimes, polysaccharide has probiotics inhibitory action, therefore selects suitable extracting mode extremely important.
Radix astragali is the pure natural product that the common people often eat, and folklore blurting out for " often drinking astragali seu Hedysari Decoction, disease-prevention health health "
Slip, mean through common radix astragali decoct soup or it is soaked on behalf of the tea, there is good disease-prevention health to act on.Radix astragali and ginseng belong to benefit
Gas good medicine, recuperating depleted yang, acute disease, the effect such as be usually used in collapsing, suffer a shock are preferable.And radix astragali is usually used in body and declines then based on qi-restoratives
With the passing of time, speech is low, small and weak pulse person.Radix astragali is had been used in external preparation for skin, is also had to cooperate with probiotics and is used in feed
Report.
The invention has the beneficial effects that：
1. the method for the present invention is embedded probiotics by microcapsules, the anti-oxidant of probiotics itself, antiacid energy are improved
Power, so as to extend the room temperature storage life of viable bacteria, improves viable bacteria ordinary temperature stability.
2. after present invention compounding nostoc extracting solution, survival rate height, the good probiotic microcapsule of envelop rate can be obtained.
3rd, nostoc contains a large amount of benefit materials, especially nostoc polysaccharide in itself, there is good promotion to epithelial cell
Palingenesis, and probiotics have collaboration cosmetic result together.
Embodiment
Embodiment 1：
A kind of preparation method of external application probiotic microcapsule, its step are as follows：
(1), by Bifidobacterium, lactobacillus acidophilus, Lactobacillus rhamnosus lyophilized bacterium powder, be inoculated in respectively in triangular flask living
Change is handled, i.e., after 30 DEG C culture 60h, expands fermented and cultured, when its zymotic fluid viable bacteria concentration reaches 1010Cfu/g, terminates fermentation,
By the zymotic fluid of three kinds of bacterium according to volume ratio 3：3：4 are mixed, and obtain bacteria suspension；
（2）, take fresh nostoc, crush, radiated 5 minutes with microwave power 280W, add the boiling boiling 20min of 5 times of weight,
Filter while hot, continue to decoct to water is added in the dregs of a decoction, repeat extraction three times, merging filtrate, refined filtration, obtains nostoc extracting solution；
（3）, by bacteria suspension, nostoc extracting solution and sodium alginate soln are 1 by volume：1：8 are mixed, and are added
Enter the Tween 80 that weight fraction is 0.4%, after emulsifying 18 minutes, sprayed into emulsion as magnetic agitation with sterile drive nozzle
In calcium chloride solution on device, after curing 12 minutes, after wet capsule is formed, with filtered through gauze, then rushed again with physiological saline
Wash 3 times, initial microcapsules preparation condition is sodium alginate 2%, and calcium chloride 2%, emulsification and immobilization time are 15min.
The percentage is mass fraction.
Embodiment 2：
A kind of preparation method of external application probiotic microcapsule, its step are as follows：
(1), by Bifidobacterium, lactobacillus acidophilus, Lactobacillus rhamnosus lyophilized bacterium powder, be inoculated in respectively in triangular flask living
Change is handled, i.e., after 35 DEG C culture 50h, expands fermented and cultured, when its zymotic fluid viable bacteria concentration reaches 1010Cfu/g, terminates fermentation,
By the zymotic fluid of three kinds of bacterium according to volume ratio 3：3：4 are mixed, and obtain bacteria suspension；
（2）, take fresh nostoc, crush, radiated 3 minutes with microwave power 320W, add the boiling boiling 25min of 5 times of weight,
Filter while hot, continue to decoct to water is added in the dregs of a decoction, repeat extraction three times, merging filtrate, refined filtration, obtains nostoc extracting solution；
（3）, by bacteria suspension, nostoc extracting solution and sodium alginate soln are 1 by volume：1：8 are mixed, and are added
Enter the Tween 80 that weight fraction is 0.4%, emulsification after twenty minutes, with sterile drive nozzle sprays into emulsion as magnetic agitation
In calcium chloride solution on device, cure after ten minutes, after wet capsule is formed, with filtered through gauze, then rushed again with physiological saline
Wash 3 times, initial microcapsules preparation condition is sodium alginate 2%, and calcium chloride 2%, emulsification and immobilization time are 15min.
The percentage is mass fraction.
Embodiment 3：
A kind of preparation method of external application probiotic microcapsule, its step are as follows：
(1), by Bifidobacterium, lactobacillus acidophilus, Lactobacillus rhamnosus lyophilized bacterium powder, be inoculated in respectively in triangular flask living
Change is handled, i.e., after 33 DEG C culture 59h, expands fermented and cultured, when its zymotic fluid viable bacteria concentration reaches 1010Cfu/g, terminates fermentation,
By the zymotic fluid of three kinds of bacterium according to volume ratio 3：3：4 are mixed, and obtain bacteria suspension；
（2）, take fresh nostoc, crush, radiated 4 minutes with microwave power 300W, add the boiling boiling 21min of 5 times of weight,
Filter while hot, continue to decoct to water is added in the dregs of a decoction, repeat extraction three times, merging filtrate, refined filtration, obtains nostoc extracting solution；
（3）, by bacteria suspension, nostoc extracting solution and sodium alginate soln are 1 by volume：1：8 are mixed, and are added
Enter the Tween 80 that weight fraction is 0.4%, after emulsifying 19 minutes, sprayed into emulsion as magnetic agitation with sterile drive nozzle
In calcium chloride solution on device, after curing 11 minutes, after wet capsule is formed, with filtered through gauze, then rushed again with physiological saline
Wash 3 times, initial microcapsules preparation condition is sodium alginate 2%, and calcium chloride 2%, emulsification and immobilization time are 15min.
The percentage is mass fraction.
Embodiment 4：
A kind of preparation method of external application probiotic microcapsule, its step are as follows：
(1), by Bifidobacterium, lactobacillus acidophilus, Lactobacillus rhamnosus lyophilized bacterium powder, be inoculated in respectively in triangular flask living
Change is handled, i.e., after 31 DEG C culture 51h, expands fermented and cultured, when its zymotic fluid viable bacteria concentration reaches 1010Cfu/g, terminates fermentation,
By the zymotic fluid of three kinds of bacterium according to volume ratio 3：3：4 are mixed, and obtain bacteria suspension；
（2）, take fresh nostoc, crush, radiated 3 minutes with microwave power 319W, add the boiling boiling 24min of 5 times of weight,
Filter while hot, continue to decoct to water is added in the dregs of a decoction, repeat extraction three times, merging filtrate, refined filtration, obtains nostoc extracting solution；
（3）, by bacteria suspension, nostoc extracting solution and sodium alginate soln are 1 by volume：1：8 are mixed, and are added
Enter the Tween 80 that weight fraction is 0.4%, after emulsifying 18 minutes, sprayed into emulsion as magnetic agitation with sterile drive nozzle
In calcium chloride solution on device, after curing 12 minutes, after wet capsule is formed, with filtered through gauze, then rushed again with physiological saline
Wash 3 times, initial microcapsules preparation condition is sodium alginate 2%, and calcium chloride 2%, emulsification and immobilization time are 15min.
The percentage is mass fraction.
Embodiment 5：
A kind of preparation method of external application probiotic microcapsule, its step are as follows：
(1), by Bifidobacterium, lactobacillus acidophilus, Lactobacillus rhamnosus lyophilized bacterium powder, be inoculated in respectively in triangular flask living
Change is handled, i.e., after 34 DEG C culture 57h, expands fermented and cultured, when its zymotic fluid viable bacteria concentration reaches 1010Cfu/g, terminates fermentation,
By the zymotic fluid of three kinds of bacterium according to volume ratio 3：3：4 are mixed, and obtain bacteria suspension；
（2）, take fresh nostoc, crush, radiated 5 minutes with microwave power 310W, add the boiling boiling 22min of 5 times of weight,
Filter while hot, continue to decoct to water is added in the dregs of a decoction, repeat extraction three times, merging filtrate, refined filtration, obtains nostoc extracting solution；
（3）, by bacteria suspension, nostoc extracting solution and sodium alginate soln are 1 by volume：1：8 are mixed, and are added
Enter the Tween 80 that weight fraction is 0.4%, emulsification after twenty minutes, with sterile drive nozzle sprays into emulsion as magnetic agitation
In calcium chloride solution on device, cure after ten minutes, after wet capsule is formed, with filtered through gauze, then rushed again with physiological saline
Wash 3 times, initial microcapsules preparation condition is sodium alginate 2%, and calcium chloride 2%, emulsification and immobilization time are 15min.
The percentage is mass fraction.
Embodiment 6：
(1) embedding rate, survival rate measure
Embed calculation of yield method：By the microcapsules of preparation with taking the wet capsules of 1g to be placed in 9mL after brine 3 times
Solve in cyst fluid, vibrated in 37 ± 1 DEG C of waters bath with thermostatic control, handle 45min, count plate, acquired results are multiplied by the gross weight of wet capsule
Measure and be denoted as N0, the viable count in bacterium glue mixed liquor before microencapsulation is denoted as N.
Embed yield=N0/N × 100%
Bacteria suspension is made with sterile saline dissolving respectively in zymotic fluid and microcapsules solution cyst fluid, then by it with 10 times
Gradient dilution, then it is 10 to take dilution factor respectively- 7To 10- 8Bacteria suspension 1mL be inoculated in deep agar culture medium count .37
DEG C culture 2d, observation counting take its average value, and are scaled the viable count of unit volume, investigate each factor to the micro- glue of Bifidobacterium
Capsule viable count survival rate influence conversion viable count formula be：The every milliliter of original bacteria liquid viable count=repetition of same dilution factor 2
Plate bacterium colony average × extension rate × 10, calculates every milliliter of original bacteria liquid total viable count.
Based on the condition of embodiment 1, the preparation and addition of nostoc extracting solution are omitted, other conditions are all identical,
Prepare the microcapsules of control group.
Experimental result is as shown in table 1 below：
Table 1
Can significantly it find out from upper table, microcapsules of the invention have very outstanding embedding rate and survival rate, and
It is significantly higher than control group.This explanation, after compounding nostoc extracting solution, the embedding to viable bacteria plays unexpected promotion effect
Fruit.In addition, nostoc also has good cosmetology effect in itself, in external preparation for skin,
（2）Purposes
The present invention capsule can with external application, including immediately dissolving after external application, can also take orally.
Practice have shown that external application has a good skin-care effect, especially in terms of antiallergic sense, for acute skin allergy, 24
It can recover normal completely in hour.
The foregoing is only a preferred embodiment of the present invention, but protection scope of the present invention be not limited thereto,
Any one skilled in the art the invention discloses technical scope in, technique according to the invention scheme and its
Inventive concept is subject to equivalent substitution or change, should be covered by the protection scope of the present invention.
Claims (1)
1. a kind of preparation method of the antianaphylactic probiotic microcapsule of external application, its step are as follows：
（1）, by Bifidobacterium, lactobacillus acidophilus, Lactobacillus rhamnosus lyophilized bacterium powder, be inoculated in respectively in triangular flask at activation
Reason, i.e., after 30~35 DEG C are cultivated 50~60h, expand fermented and cultured, when its zymotic fluid viable bacteria concentration reaches 1010Cfu/g, terminates hair
Ferment, by the zymotic fluid of three kinds of bacterium according to volume ratio 3：3：4 are mixed, and obtain bacteria suspension；
（2）, take fresh nostoc, crush, radiated 3-5 minute with microwave power 280-320W, add the boiling of 5 times of weight to boil 20-
25min, filters while hot, continues to decoct to water is added in the dregs of a decoction, repeats extraction three times, merging filtrate, refined filtration, obtains nostoc and carry
Take liquid；
（3）, by bacteria suspension, nostoc extracting solution and sodium alginate soln by volume be 1：1：8 are mixed, and are added dense
After the Tween 80 spent for 0.4% emulsifies 18-20 minutes, emulsion is sprayed into as on magnetic stirring apparatus with sterile drive nozzle
In calcium chloride solution, after curing 10-12 minutes, after wet capsule is formed, with filtered through gauze, normal saline flushing 3 is then used again
Secondary, the sodium alginate soln concentration is 2 %, and the concentration of calcium chloride solution is 2 %, and emulsification and immobilization time are 15
Minute.
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| CN111067878A (en) * | 2019-12-30 | 2020-04-28 | 武汉轻工大学 | Ginsenoside-bifidobacterium breve M-16V microcapsule and preparation method thereof |
| CN112754988B (en) * | 2021-03-22 | 2022-06-28 | 中南大学湘雅二医院 | Probiotic skin care product capable of reducing skin sensitization and preparation method thereof |
| CN114246340A (en) * | 2021-12-22 | 2022-03-29 | 湖南炎帝生物工程有限公司 | Nostoc sphaeroids kutz probiotic composition and preparation method and application thereof |
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