CN104586738B - Tuber of dwarf lilyturf liquid fermentate, cosmetics containing fermentate and its production and use - Google Patents
Tuber of dwarf lilyturf liquid fermentate, cosmetics containing fermentate and its production and use Download PDFInfo
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Abstract
The invention belongs to biological fermentation field, and in particular to tuber of dwarf lilyturf liquid fermentate, cosmetics containing fermentate and its production and use.Technical problem solved by the invention is that, to instruct, providing one kind using the liquid fermentation system of the tuber of dwarf lilyturf and lactic acid bacteria has replenishing water and preserving moisture, the product of anti-ageing effect with " bioconversion is theoretical ".Tuber of dwarf lilyturf liquid fermentate of the invention is to add the tuber of dwarf lilyturf in the medium, then with the fermented liquid mixture obtained by lactobacillus-fermented；Wherein, in culture medium in terms of crude drug weight, the tuber of dwarf lilyturf that percentage by weight is 5~20% is contained；The inoculum concentration of lactic acid bacteria is 1.0 × 104~1.0 × 108cfu/ml。
Description
Technical field
The invention belongs to biological fermentation field, and in particular to tuber of dwarf lilyturf liquid fermentate, the cosmetics containing fermentate and its system
Preparation Method and purposes.
Background technology
The tuber of dwarf lilyturf is medicinal material simply in traditional chinese medicine, first recorded in the Eastern Han Dynasty《Sheng Nong's herbal classic》.Advised in current China pharmacopeia
The fixed medicinal tuber of dwarf lilyturf is the dried root of the liliaceous plant tuber of dwarf lilyturf.The Chinese Pharmacopoeia record tuber of dwarf lilyturf has nourishing Yin and promoting production of body fluid, moistening lung clears away heart-fire
The effects such as.Tuber of dwarf lilyturf main chemical compositions are polysaccharide, steroid saponin, homoisoflavone class, amino acid etc.；Ophiopogon japonicus extract is being made up
Used in product frequently as moisturizing additive.
Lactic acid bacteria is a kind of probiotics being present in mankind's body, be fermenting carbohydrate primary product be lactic acid a class without bud
The general name of spore, gram-positive bacterium, the overwhelming majority is all bacterium essential in human body and with important physiological function
Group, is widely present in the enteron aisle of human body.The primary product of lactobacillus-fermented is lactic acid, and lactic acid is acid, therefore can be answered
For being used for the regulation of ph values in cosmetic industry；Simultaneously as lactic acid is the composition portion of NMF NMF in skin
Point, therefore, moisturizing is an important use of the lactic acid in cosmetics.Lactic acid after general neutralization is used as moisturizing ingredient.
The present invention is intended to provide a kind of with replenishing water and preserving moisture, the ophiopogon liquid as obtained by the tuber of dwarf lilyturf and lactobacillus-fermented of anti-ageing effect
Body fermentate.
The content of the invention
Technical problem solved by the invention is to instruct, using the tuber of dwarf lilyturf and the liquid of lactic acid bacteria with " bioconversion is theoretical "
Fermentation system provides a kind of with replenishing water and preserving moisture, the product of anti-ageing effect.
First technical problem solved by the invention is to provide the ophiopogon liquid obtained by the liquid fermentation of the tuber of dwarf lilyturf and lactic acid bacteria
Body fermentate.
Tuber of dwarf lilyturf liquid fermentate of the invention, culture medium is constituted with the tuber of dwarf lilyturf and water, then the liquid obtained by inoculating lactic acid bacterium fermentation
Body fermenting mixture；Wherein, culture medium is in terms of crude drug weight, and tuber of dwarf lilyturf percentage by weight is 5~20%；The inoculum concentration of lactic acid bacteria is
1.0×104~1.0 × 108cfu/ml。
In above-mentioned technical proposal, culture medium is in terms of crude drug weight, and preferably tuber of dwarf lilyturf percentage by weight is 10%.The tuber of dwarf lilyturf is primary
Medicinal material or tuber of dwarf lilyturf medicine materical crude slice.
In above-mentioned technical proposal, the inoculum concentration of lactic acid bacteria is preferably 1.0 × 106cfu/ml。
Second technical problem solved by the invention is to provide the preparation method of tuber of dwarf lilyturf liquid fermentate of the invention, including
Following steps：
A, with the tuber of dwarf lilyturf and water composition culture medium, in terms of crude drug weight, tuber of dwarf lilyturf percentage by weight be 5~20%；
B, inoculating lactic acid bacterium, inoculum concentration are 1.0 × 104~1.0 × 108cfu/ml；
C, culture 1~5 day under conditions of 30 DEG C~40 DEG C and 0~200rpm, produce tuber of dwarf lilyturf liquid fermentate.
In above-mentioned technical proposal, the preferred addition of culture medium is in terms of crude drug weight, and tuber of dwarf lilyturf percentage by weight is 10%.
In above-mentioned technical proposal, the inoculum concentration of lactic acid bacteria is preferably 1.0 × 106cfu/ml。
In above-mentioned technical proposal, cultivated 5 days under conditions of 37 DEG C and 50rpm.
In above-mentioned technical proposal, the lactic acid bacteria culturers that inoculation is used can be selected using at present by various usual way cultures
Obtained strain, is such as used for the lactobacillus inoculation for manufacturing pickles, Yoghourt.
Wherein, a kind of preferred strain source and training method are：
A, lactic acid bacteria culturers are cultivated in solid medium；
B, the step A lactobacillus inoculums cultivated are entered in lactic acid bacteria fluid nutrient medium；
C, 1~5 day gained lactic acid bacteria culturers of culture under conditions of 30 DEG C~40 DEG C and 0~200rpm.
In above-mentioned technical proposal, carbon source, nitrogen containing lactic acid bacteria normal fermentation in lactic acid bacteria fluid nutrient medium described in step B
Source, trace element and/or inorganic salts ingredients, wherein：Carbon source composition is the 0.5%~10% of fluid nutrient medium gross weight, nitrogen source
Composition is 0.5%~3%, and trace element and/or inorganic salts ingredients are 0.01%~1%, and remaining is water, sterilizing.Wherein
Described carbon source, nitrogen source can select the conventional composition in current collective media, such as glucose, sucrose, starch, beef
The conventional medium constituent such as cream, yeast extract.Further preferably described sterilizing is through 115~121 DEG C of 15~30min of sterilizing.
In above-mentioned technical proposal, solid medium described in step A is by the lactic acid bacteria fluid nutrient medium described in step B
1000ml and 10~20g of agar compositions, conventional sterilant processing is made by the same manner.Condition of culture is：In 30-40 DEG C and 0-
Cultivated 1-3 days under conditions of 200rpm；It is preferred that condition of culture is：Cultivated 1 day under conditions of 37 DEG C and 50rpm.
3rd technical problem solved by the invention is to provide the purposes of tuber of dwarf lilyturf liquid fermentate of the invention, specific manifestation
Having for tuber of dwarf lilyturf liquid fermentate of the invention significantly has replenishing water and preserving moisture, anti-ageing effect.
4th technical problem solved by the invention is to provide using tuber of dwarf lilyturf liquid fermentate of the invention or its extract
For active component, the cosmetics that acceptable auxiliary material is prepared from cosmetics are added.
Cosmetics prepared by the present invention can be the common cosmetics such as toner, emulsion, Essence, face cream, facial mask, eye mask
Product.
It is used for the nursing of skin, more single ophiopogon japonicus extract and lactobacillus-fermented using tuber of dwarf lilyturf liquid fermentate of the invention
For thing, with good hygroscopicity and moisture-keeping function, so as to play a part of replenishing water and preserving moisture；To DPPH free radicals and hydroxyl certainly
There is stronger scavenging action by base, the oxidative damage that human keratinized cell is subject to can be prevented and treated, with good antioxidation,
So as to possess clear and definite anti-ageing effect.The cosmetics that tuber of dwarf lilyturf liquid fermentate of the invention is prepared according to a conventional method also have clear and definite
Replenishing water and preserving moisture and anti-ageing effect.
Embodiment
The embodiment of form, remakes further specifically to the above of the invention by the following examples
It is bright, illustrate but do not limit the present invention.
The lactic acid bacteria of embodiment 1 is tested the growth adaptability of the fluid nutrient medium containing tuber of dwarf lilyturf medicinal material and influence
(1) 1.0 × 10 are pressed5Lactic acid bacteria is inoculated in containing 5.0% by cfu/ml inoculum concentrations respectively, 10.0%, 15.0%,
In the fluid nutrient medium of 20.0% tuber of dwarf lilyturf (in terms of crude drug weight, as follows), 3~5 are cultivated under the conditions of 37 DEG C and 50rpm
My god, the lactic acid bacteria clump count in zymotic fluid is detected using spread plate.As a result it is as shown in table 1.
The result of table 1 shows that, when tuber of dwarf lilyturf content is 10.0%, the growing way of lactic acid bacteria is good, and basically identical；When tuber of dwarf lilyturf water extraction
The concentration of liquid is more than after 10%, and the growth of lactic acid bacteria is suppressed.Therefore in liquid fermentation base, the optimum content of the tuber of dwarf lilyturf is
10%.
The influence that table 1 tuber of dwarf lilyturf content and fermentation time are cultivated lactic acid bacteria
Extension with incubation time can also be seen that by the result of table 1, the clump count of lactic acid bacteria gradually increases, from production
Cost consideration, it is determined that the fermentation time of lactic acid bacteria is 120h, i.e., 5 day in the present invention.
(2) 1.0 × 10 are pressed4、1.0×105With 1.0 × 106Lactic acid bacteria is inoculated in containing 10% by cfu/ml inoculum concentrations respectively
In the fluid nutrient medium of the tuber of dwarf lilyturf, cultivated 5 days under the conditions of 37 DEG C and 50rpm, the breast in zymotic fluid is detected using spread plate
Sour bacterium clump count.As a result it is as shown in table 2
The influence that the inoculum concentration of table 2 is cultivated lactic acid bacteria
Viable count in zymotic fluid it can be seen from the result of table 2 increases with the increase of inoculum concentration, accordingly, it is determined that this
Inoculum concentration in invention is 1.0 × 106cfu/ml。
The efficacy test and irritation test of the present composition of embodiment 2
Added by the above-mentioned tuber of dwarf lilyturf by terms of crude drug weight by 10.0% percentage in culture medium, by 1.0 × 106cfu/
After ml amount inoculating lactic acid bacterium, culture obtains described tuber of dwarf lilyturf liquid fermentate for 5 days under the conditions of 37 DEG C and 50rpm, under progress
The efficacy test and irritation test stated.
First, fuchsin(e)test and moisturizing are tested
Will be by saturation magnesium chloride brine as the drier of relative humidity (RH) 33% and by the saturated ammonium sulfate aqueous solution
It is put into as the drier of relative humidity (RH) 81% in the water-impermeable incubator that design temperature is 20 ± 0.1 DEG C, it is accurate respectively
Weigh 1.00g tuber of dwarf lilyturf liquid fermentate dry powder, tuber of dwarf lilyturf medicinal material dry powder, the sample of lactic acid bacteria liquid fermentate dry powder is to a diameter of
In 90mm knot product ware, then it is put into two driers, is taken out after 24h, the weight of accurate title most each sample, the sample before and after experiment
The weight difference of product, the hydroscopicity of sample is calculated with following formula：
Hydroscopicity (%)=100 × (W1-W0)/W0
W0 is weight before sampling test in formula；Wl is weight (after 24h) after sampling test.
By tuber of dwarf lilyturf liquid fermentate dry powder, tuber of dwarf lilyturf medicinal material dry powder, three samples of lactic acid bacteria liquid fermentate dry powder spend from
After sub- water is diluted to 10%, 5.00g sample aqueous solutions are accurately weighed respectively into diameter 90mm crystallising dish, then are put into two does
In dry device, taken out after 24h, the weight of precise each sample, the weight difference of sample, sample is calculated with following formula before and after experiment
Moisturizing rate：
Moisturizing rate (%)=100 × (W3-W2)/W2
W2 is weight before sampling test in formula；W3 is weight (after 24h) after sampling test.
Moisture absorption and performance of keeping humidity of each sample of table 3 under different relative humidity
Result of the test is as shown in table 3, from results showed that tuber of dwarf lilyturf fermentate in the case of two kinds of low humidity and high humidity, its
Hydroscopicity and moisturizing rate are better than unfermentable tuber of dwarf lilyturf medicinal material and single lactobacillus fermentation dry powder, illustrate by this granting
The tuber of dwarf lilyturf liquid fermentate that technology is prepared has better water suction moisture-keeping function.
2nd, oxidation resistance testing experiment
1. anti-oxidation efficacy detects sample preparation
Tuber of dwarf lilyturf liquid fermentation production dries dry powder processed, crushes；Tuber of dwarf lilyturf pulverizing medicinal materials；Lactic acid bacteria liquid fermentate drying system is dry
Powder, is crushed；Above-mentioned fine powder crosses 60 mesh sieves, takes powder 5g to be dissolved in after 100ml water, 60 DEG C of ultrasonic extraction 1h, 3000g centrifugations
10min, filtering and collecting filter liquid constant volume is that 5ml, i.e. concentration are 1g/ml, is then diluted to 40-400mg/ml.
2. remove the measure of OH free radical abilities
The phosphate buffer solution (0.05mol/L) that 1mlpH values are 7.4,0.4ml 6mmol/L are separately added into test tube
Phen solution (absolute ethyl alcohol preparation), fully mix after, add 0.4ml 6mmol/L FeSO4 solution, after addition stand
Mix.Then the certain density sample solutions of 0.5ml are added thereto, are mixed, are added 0.5ml 0.1% H2O2, finally
Volume is supplemented to 4ml, while doing blank test.It is another to do damage again and do not damage pipe.Wherein 0.5ml 0.1% is added in damage pipe
H2O2, pipe is not damaged is not added with H2O2, each volume is finally supplemented to 4ml.In the extinction being incubated at 37 DEG C under 1h, measurement 536nm
Degree, is repeated twice, and calculates average value.Scavenging action to hydroxyl free radical=(Ai-A)/(A0- A) × 100%.
A is the light absorption value of solution when being not added with sample, A0Not to be loaded and H2O2When solution light absorption value.Notice that concentration is controlled,
3-4 times of concentration is first tested, then is progressively diluted to optimum concentration, temperature is controlled.
3. the measure of extract scavenging ability of DPPH free radical
Testing sample 1ml and the 0.4mg/ml prepared in advance with 95% ethanol DPPH free radical reagents 1ml are taken in 10ml
Teat glass in.Sample vibrates after adding determines its light absorption value (A after 30s, 37 DEG C of insulation 10min under 517nm wavelengthi).Together
When determine and be not added with the sample blank light absorption value (A of DPPH free radical reagentsj) and add DPPH free radicals reagent but be not added with the suction of sample
Light value (A0), it is as shown in table 4 below.
The DPPH of table 4 experiment sample-adding tables
Sample to DPPH removing percentage I=1- [(Ai-Aj)/A0] × 100%,
Result of the test：The EC50 of sample is extrapolated by clearance rate.As a result it is as shown in table 5.
The sample of table 5 removes the EC of OH, DPPH free radical50Result of the test
As shown in table 5, in terms of OH, DPPH free radical is removed, the EC50 of tuber of dwarf lilyturf fermentate is below wheat to result of the test
Winter medicinal material and lactobacillus fermentation, illustrate that its antioxidant effect is good, with more preferable radical scavenging activity.
3rd, to H2O2The anti-ageing efficacy test experiment of the model of oxidative of the human epidermal cell of induction
Using the DEME medium culture human epidermal cell HaCaT cells containing 10% hyclone, tryptose during passage
Enzyme is digested.During by cell culture to exponential phase of growth, it is inoculated in 96 orifice plates, inoculum density is 6 × 103/hole, is incubated
After 24h, it is loaded by following packet：Blank control group, H2O2 damage models group, the basic, normal, high dosage group of tuber of dwarf lilyturf fermentate (25,
50,100 μ g/ml), tuber of dwarf lilyturf medicinal material group (50 μ g/ml), lactobacillus fermentation group (50 μ g/ml)；3 multiple holes are set；Continue to be incubated
48h.6h before culture is terminated, in addition to blank control group, other each group cells are added after 100mM H2O2, effect 2h, are sucked per hole
Culture medium, adds 100 μ l PBS, 10mg/ml MTT, is incubated after 4h, adds the 10%SDS terminating reactions of 0.1M HCl acidifyings.
After 4h, OD values are measured in 570nm, survival rate of each group cell relative to model cell is calculated.
The sample of table 6 is to H2O2The preventive and therapeutic effect of caused cell oxidative damage
As shown in table 6, tuber of dwarf lilyturf medicinal material and lactobacillus fermentation can be to a certain extent to people angles caused by H2O2 for result of the test
Cell plastid oxidative damage plays preventive and therapeutic effect；But (50 μ g/ under the tuber of dwarf lilyturf fermentate obtained after the two is fermented, its comparable sodium
Ml), its prophylactic-therapeutic effect to human keratinized cell oxidative damage caused by H2O2 is better than tuber of dwarf lilyturf medicinal material and single lactobacillus fermentation；
In terms of three concentration tests results of tuber of dwarf lilyturf fermentate, illustrate that tuber of dwarf lilyturf fermentate has obvious anti-oxidant, anti-aging effects, and
There is significant dose-effect relationship.
4th, tuber of dwarf lilyturf fermentate safety testing
Using " acute oral and the acute dermal toxicity experiment " in current cosmetic material detection and " skin and urgency
Property eye irritation/corrosion test ", result of the test shows that tuber of dwarf lilyturf fermentate belongs to nontoxic, nonirritant, non-corrosiveization
Cosmetic raw material, available in the formula of cosmetics.
There is the characteristic of good moisturizing water suction from above-mentioned results showed that tuber of dwarf lilyturf liquid fermentate, its moisturizing rate is inhaled
Wet rate is superior to unfermentable tuber of dwarf lilyturf medicinal material and the lactobacillus fermentation of single fermentation；Tuber of dwarf lilyturf liquid fermentate has substantially simultaneously
Inoxidizability, to H2O2The oxidative damage of caused human keratinized cell has good protection and preventive and therapeutic effect；And in comparable sodium
Down better than unfermentable tuber of dwarf lilyturf medicinal material and the lactobacillus fermentation of single fermentation, point out tuber of dwarf lilyturf liquid fermentate that there is very strong benefit
Water moisturizing and anti-oxidant, anti-aging effect.It is raw material by tuber of dwarf lilyturf liquid fermentate, its fermentate stoste or fermentation can be used
Thing powder, the cosmetics of the form such as the toner prepared according to a conventional method, emulsion, Essence, face cream, facial mask, eye mask should also have
Clear and definite replenishing water and preserving moisture, antioxidant and anti-aging effect.
Claims (14)
1. tuber of dwarf lilyturf liquid fermentate, it is characterised in that：With the tuber of dwarf lilyturf and water composition culture medium, then obtained by inoculating lactic acid bacterium fermentation
Liquid fermentation mixture；Wherein, culture medium is in terms of crude drug weight, and tuber of dwarf lilyturf percentage by weight is 5~20%；
Preparation method comprises the following steps：
A, with the tuber of dwarf lilyturf and water composition culture medium, in terms of crude drug weight, tuber of dwarf lilyturf percentage by weight be 5~20%；
B, inoculating lactic acid bacterium, inoculum concentration are 1.0 × 104~1.0 × 108cfu/ml；
C, culture 1~5 day under conditions of 30 DEG C~40 DEG C and 0~200rpm, produce tuber of dwarf lilyturf liquid fermentate.
2. tuber of dwarf lilyturf liquid fermentate according to claim 1, it is characterised in that：Culture medium is in terms of crude drug weight, tuber of dwarf lilyturf weight
It is 10% to measure percentage.
3. tuber of dwarf lilyturf liquid fermentate according to claim 1, it is characterised in that：The tuber of dwarf lilyturf is to drink primary crude drug or the tuber of dwarf lilyturf
Piece.
4. the tuber of dwarf lilyturf liquid fermentate according to claim any one of 1-3, it is characterised in that：Lactic acid bacteria connects in culture medium
The amount of kind is 1.0 × 106cfu/ml。
5. the preparation method of tuber of dwarf lilyturf liquid fermentate, it is characterised in that：Comprise the following steps：
A, with the tuber of dwarf lilyturf and water composition culture medium, in terms of crude drug weight, tuber of dwarf lilyturf percentage by weight be 5~20%；
B, inoculating lactic acid bacterium, inoculum concentration are 1.0 × 104~1.0 × 108cfu/ml；
C, culture 1~5 day under conditions of 30 DEG C~40 DEG C and 0~200rpm, produce tuber of dwarf lilyturf liquid fermentate.
6. the preparation method of tuber of dwarf lilyturf liquid fermentate according to claim 5, it is characterised in that：At least meet following any
One：
Culture medium is in terms of crude drug weight, and tuber of dwarf lilyturf percentage by weight is 10%；
The inoculum concentration of lactic acid bacteria is 1.0 × 106cfu/ml；
Cultivated 5 days under conditions of 37 DEG C and 50rpm.
7. the preparation method of tuber of dwarf lilyturf liquid fermentate according to claim 5, it is characterised in that：The culture of lactic acid bacteria culturers
Method comprises the following steps：
A, lactic acid bacteria culturers are cultivated in solid medium；
B, the step A lactobacillus inoculums cultivated are entered in lactic acid bacteria fluid nutrient medium；
C, 1~5 day gained lactic acid bacteria culturers of culture under conditions of 30 DEG C~40 DEG C and 0~200rpm.
8. the preparation method of tuber of dwarf lilyturf liquid fermentate according to claim 7, it is characterised in that：At least meet following any
One：
Carbon source, nitrogen source, trace element and/or nothing containing lactic acid bacteria normal fermentation in lactic acid bacteria fluid nutrient medium described in step B
Machine salt component；
Solid medium described in step A is by 10~20g of lactic acid bacteria fluid nutrient medium 1000ml and agar described in step B
Composition.
9. the preparation method of tuber of dwarf lilyturf liquid fermentate according to claim 8, it is characterised in that：Carbon source composition in culture medium
For the 0.5%~10% of fluid nutrient medium gross weight, nitrogen source composition is 0.5%~3%, trace element and/or inorganic salts ingredients
For 0.01%~1%, remaining is water.
10. the preparation method of tuber of dwarf lilyturf liquid fermentate according to claim 8, it is characterised in that：Step A condition of culture
For：Cultivated 1-3 days under conditions of 30-40 DEG C and 0-200rpm.
11. the preparation method of tuber of dwarf lilyturf liquid fermentate according to claim 10, it is characterised in that：Step A condition of culture
For：Cultivated 1 day under conditions of 37 DEG C and 50rpm.
12. any one of the claim 1-4 tuber of dwarf lilyturf liquid fermentates prepare have replenishing water and preserving moisture, the medicine of anti-ageing effect or
Applied in cosmetics.
13. cosmetics, it is characterised in that：Using any one of the claim 1-4 tuber of dwarf lilyturf liquid fermentates or its extract as work
Property composition, add the cosmetics that are prepared from of acceptable auxiliary material on cosmetics.
14. cosmetics according to claim 13, it is characterised in that：The cosmetics be toner, emulsion, Essence,
Face cream, facial mask, eye mask.
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| CN109350584B (en) * | 2018-10-17 | 2022-12-13 | 广州市孝真生物科技有限公司 | Fermented raw juice of traditional Chinese medicine composition with powerful moisturizing effect and preparation and application thereof |
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| CN101633875A (en) * | 2009-02-22 | 2010-01-27 | 王怀能 | Biofermentation method of dwarf lilyturf wine |
| KR101226758B1 (en) * | 2010-09-28 | 2013-01-25 | 한국 한의학 연구원 | Composition for Prevention or Treatment of Atopic Dermatitis Comprising Herbal Extract and Fermentation Product thereof with Lactic acid Bacteria |
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