CN105901741B - A kind of dietary supplements with the probiotics and plant extracts of alleviating lead poisoning sexual function - Google Patents
A kind of dietary supplements with the probiotics and plant extracts of alleviating lead poisoning sexual function Download PDFInfo
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- CN105901741B CN105901741B CN201610168450.0A CN201610168450A CN105901741B CN 105901741 B CN105901741 B CN 105901741B CN 201610168450 A CN201610168450 A CN 201610168450A CN 105901741 B CN105901741 B CN 105901741B
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- dietary supplements
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- lead
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
Landscapes
- Medicines Containing Plant Substances (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of dietary supplements with the probiotics and plant extracts of alleviating lead poisoning sexual function, belong to food technology field.The present invention is that this plant of bacterium is aided with to plant extracts using the bacterium powder for being freeze-dried acquisition as main ingredient, and the probiotics of tablet form and the dietary supplements of plant extracts are prepared with direct powder compression after adding proper auxiliary materials, it contains viable bacteria content greater than 109The lactobacillus plantarum CGMCC 5494 of CFU/g.The dietary supplements obtained in this way can reduce lead content in lead exposure Mice Body；Restore oxidativestress damage caused by lead exposure；Improve the activity of aminolevulinate dehydratase in lead exposure Mice Body；Reduce zinc protoporphyrin content in lead exposure Mice Body；Restore the intellectual damage of lead exposure mouse.It has a vast market foreground.
Description
Technical field
The present invention relates to a kind of dietary supplements with the probiotics and plant extracts of alleviating lead poisoning sexual function, belong to
Food technology field.
Background technique
Lead is a kind of silvery white heavy metal with blue, oxidizable in air, dimmed.Lead is widely used, both can be with
Effectively stop X-ray and radioactive ray, and the main material of manufacture battery.In addition, the compound of lead is many pigment
Raw material.Lead contamination refers to compound pollution caused by ambient enviroment of lead or lead, and concentration and chemical form endanger journey with it
It spends directly related.The strong feature of lead enriching makes it be difficult to degrade in the environment, cannot pass through the catharsis of environment itself
It reduces, exceeds normal range (NR), can not only be detrimental to health, also result in environmental quality deterioration.Lead content increases in environment
The main reason for being the generation of lead contamination.Lead content in China's atmospheric environment is up to fifty-five million ton.High lead content in air
It on the one hand is caused by natural cause, such as volcano eruption, on the other hand forest fire etc. is then due to industrial pollution and traffic
Exhaust emission.Furthermore also extremely serious the phenomenon that Lead Pollution in Soil.After sewage irrigation soil, heavy metals exceeding standard that soil contains
Directly contribute heavy metal pollution of soil.The cereal of this soil output and vegetables, content of beary metal can be significantly exceeded.National soil
Background value basic statistics amount the result shows that, China's Content of Pb in Soil reaches as high as 1143 μ g/g, minimum 0.68 μ g/g, average
It can reach 26 μ g/g.Food, most of lead contamination of drinking water is all extraneous contamination.Leaded food, drinking water after peptic digest,
Adult absorbs up to 11%, and children absorb and are up to 30%-75%.After people drinks this tap water, lead is lodged in vivo, directly
Health is damaged.
Lead can enter human body by multiple channel, and intracorporal lead content seldom also can cause to damage to human body.Even if controlling
Decline intracorporal lead level after treatment, but the damage that has resulted in of lead be also it is permanent, can not restore as former state.Current grinds
Study carefully discovery lead poisoning to be mainly reflected in the following aspects to the toxic action of human body: A influences nervous system, be mainly shown as
Damage to intellectual aspect；B damages hematological system, and main cash is anaemia；C influences urinary system, is mainly shown as to kidney
The damage of tubule；D damages cardiovascular system, is mainly shown as the diseases such as myocarditis.
The drug of traditional two kinds treatment lead poisonings is while being effectively discharged intracorporal lead in the market, bring side effect
Also extremely serious.EDTA metalloid chelating agent calcium disodium edetate (CaNa2EDTA), to mucous membrane, the upper respiratory tract, eyes, skin
Irritating effect, and there is serious renal toxicity, other the essential trace elements of the human bodys have also been discharged while driving lead.DMSA
Class competitiveness antidote dimercaptosuccinic acid (DMSA) is possible to that midfield as above is caused to go up painful after enteron aisle is absorbed into human body,
Certain side effects such as nausea.Furthermore there is serious hepatic disorder person that cannot use DMSA.
Therefore make great efforts to explore safer, more efficient while again economic method alleviation lead poisoning to be very necessary.
Especially for the children also in the growth stage, very serious toxic side effect can be had to children by chelating agent therapy, safely and effectively
Therapy is particularly important.By the synergy of probiotics and nutrient, can to avoid or reduce the dosage of chelating agent,
The side effect for reducing chelating agent is more beneficial to health more suitable for medication.
Lactic acid bacteria is one kind of probiotics, is one of intracorporal necessary flora of people, is widely present in human body intestinal canal.Currently
Domestic and international biologist is it was demonstrated that lactic acid bacteria inhibits to cause a disease there are many important physiological function, including adjusting human body microecological balance
Bacterium, prevents and treats lactose intolerance, and antitumor, adjusting autoimmunity intervenes oxidative stress, reduction cholesterol etc..Current multinomial research
Confirm that lactic acid bacteria adsorbs the ultrahigh in efficiency of heavy metal in vitro, it is very outstanding to alleviation effect of body oxidative stress.Tea polyphenols
It is the general name of Polyphenols of Tea.Current multinomial research has shown that tea polyphenols have anti-solution to make heavy metallic salt and biological alkalosis
With the energy strongest hydroxy radical of extremely strong removing toxicity blocks lipid peroxidation process, improves the activity of human enzymes.Grape
Seed extract extracts from grape pip, cannot synthesize in human body, can effectively clear and find in lipid peroxidation product and nature
Anti-oxidant, the strongest substance of Scavenging ability, the effect of delaying aging and enhancing immunity with super strength.It is multinomial to grind
Studying carefully proves that grape seed extract can effectively prophylactic treatment lead poisoning.
The continuous transformation of nutrient health theory, ill to cure the disease, the lactobacillus micro-ecological preparation product of no treating diseases and making health-care is increasingly
It is welcome.It is more common with liquid lactic acid bacterium probiotics preparation however in lactobacillus micro-ecological preparation, and the benefit of tablet form
Raw bacterium dietary supplements is not common.Therefore tablet form is developed, has and alleviate lead poisoning effect, while can also has
Effect removes the free radical that is harmful to the human body, the probiotics of anti-aging and the dietary supplements of plant extracts, be it is a kind of very
The therapeutic scheme of health, has a vast market application prospect.
Summary of the invention
The present invention is to provide a kind of purposes of lactobacillus plantarum (Lactobacillus plantarum), is by this plant of bacterium
To be freeze-dried the bacterium powder obtained as main ingredient, it is aided with plant extracts, the probiotics of tablet form is prepared after addition proper auxiliary materials
With the dietary supplements of plant extracts, it contains viable bacteria content greater than 109The lactobacillus plantarum CGMCC 5494 of CFU/g.It should
Bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation address on November 29th, 2011
For Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, deposit number is CGMCC 5494.
Related lactobacillus plantarum CGMCC 5494 has following biological characteristics:
(1) thallus feature: Gram's staining is positive, and cell is in rod-short, and thallus is about 0.5-1.0 μm wide, 2-4 μm
Long, Cheng Dan, pairs of or chaining do not form gemma, and both ends are round；
(2) colony characteristics: forming apparent bacterium colony on MRS culture medium, and diameter is round between 0.3-2.0mm, milky white
Color, translucent, surface wettability is smooth, neat in edge, not chromogenesis；
(3) growth characteristics: the bacterial strain minimum growth temperature is 20 DEG C, and maximum growth temperature is 40 DEG C, at 30-37 DEG C of temperature
Lower growth is best, and the highest and lowest initial growth pH being resistant to is 9.0 and 2.5, and the initial pH of the most suitable growth is 6.0；The present invention
The period of delay of lactobacillus plantarum strain is relatively short, and 4h or so initially enters logarithmic growth phase, and 12h just reaches stationary phase.
A kind of dietary supplements containing profitable probliotics and plant extracts simultaneously provided by the invention, is to be freeze-dried and obtain
The probiotics bacterial powder obtained is main ingredient, is aided with plant extracts, is prepared with powder through pressed disc method after addition proper auxiliary materials；Institute
Stating probiotics is the lactobacillus plantarum (Lactobacillus plantarum) that deposit number is CGMCC 5494.
In one embodiment of the invention, the probiotics bacterial powder the preparation method is as follows: by lactobacillus plantarum
CGMCC 5494 expands culture, centrifugation, washing after activating, freeze drying protectant is then added, thallus is resuspended, reach cell concentration
1011CFU/mL or more, pre-freeze after mixing, is then freeze-dried.
In one embodiment of the invention, the freeze-drying is the trehalose or 100- with 100-150g/L
The degreasing milk solution of 250g/L is as freeze drying protectant.
In one embodiment of the invention, the freeze-drying is using the trehalose of 100g/L as frozen-dried protective
Agent.
In one embodiment of the invention, the freeze-dried vaccine powder preparation the following steps are included:
A. the preparation of culture medium: using by based on culture medium gross mass 87.7% water by 10% enzyme hydrolysis skimmed milk,
0.5% glucose, 1.5% tryptone and 0.3% yeast extract dissolve, and then adjusting its pH is 6.8, obtain culture medium；
B. it the preparation of freezing drying protective agent: is mixed with, is respectively obtained containing 100g/L using water and protective agent raw material
The protective agent of trehalose, the protective agent containing 100g/L skimmed milk power, the protective agent containing 50g/L sorbierite contain 150g/L
The protective agent of sucrose.The present invention considers the non-functional component for simplifying the dietary supplements of probiotics and plant extracts as far as possible,
The combination of a variety of freeze drying protectants is not tested, only the protecting effect of single kind freezing drying protective agent is carried out
Compare.
C. 5494 strain of lactobacillus plantarum CGMCC is inoculated into based on the quality of the culture medium with the inoculum concentration of 2-4%
At 110-120 DEG C of temperature in culture medium cooling after sterilizing 8-12min, 18h then is cultivated at 37 DEG C, with pH7.2 phosphate
Buffer solution for cleaning 2-4 times, the resuspension of the freezing drying protective agent described in step B make viable bacteria concentration up to 1011CFU/mL, allows the suspension
Liquid preculture 60min at 37 DEG C of temperature is freeze-dried to obtain the freeze-dried vaccine powder later.
In one embodiment of the invention, the dietary supplements, according to parts by weight containing the bacterium of 30-50%
Powder, the plant extracts of 4-8%, the auxiliary material of 42-66%.
In one embodiment of the invention, the plant extracts is grape seed extract；Auxiliary material is a kind of or more
Plant filler usually used in tablets, adhesive, disintegrating agent, lubricant.
The dietary supplements, according to parts by weight containing the microcrystalline cellulose of 37.9-59.7%, the low substitution of 4-6%
Hydroxypropyl cellulose, the superfine silica gel powder of 0.1-0.3%, the grape seed extract of 2.5-5%, the tea polyphenols of 1.5-3%, 30-
50% bacterium powder.
In one embodiment of the invention, it is through tablet press machine with 2.4t pressure tabletting that the pressed disc method, which carries out tabletting,
Obtained tablet weight is 250mg ± 15mg.
In one embodiment of the invention, the dietary supplements, according to parts by weight containing 48.9% crystallite
Cellulose, 5% low-substituted hydroxypropyl cellulose, 0.1% superfine silica gel powder, 3.6% grape seed extract, 2.4% tea
Polyphenol, 40% bacterium powder.Formula is as shown in table 1 below:
The dietary supplements of 1 probiotics of table and plant extracts composition and content
In one embodiment of the invention, the viable bacteria of lactobacillus plantarum CGMCC 5494 contains in the dietary supplements
Amount reaches 109CFU/g or more.
The probiotics of tablet form described in one embodiment of the present invention and the dietary supplements of plant extracts
Production method is as shown in Fig. 1.
Beneficial effects of the present invention:
(1) meal supplement tablet while being prepared for the first time containing profitable probliotics and plant extracts；
(2) dietary supplements active component content of the invention is high, shelf life is long；Prebiotic bacterial content can reach 8.3 ×
109Cfu/g, viable count is still greater than 10 after storing 6 months at 4 DEG C5CFU/g；Anthocyanidin content reach 29.17mg/g, EGC, EC,
The content of EGCG, ECG are respectively up to 2.61mg/g, 1.23mg/g, 7.59mg/g, 1.53mg/g, after storing 6 months at 20 DEG C still
There is 90% or more retention rate；
(3) dietary supplements of the invention can reduce the lead content and blood of the internal organs such as liver in Mice Body, kidney, brain
Lead content, alleviate the lead poisoning of Mice Body internal cause caused by oxidativestress damage, aminolevulinate dehydratase activity reduce, zinc
Protoporphyrin content rises, and intelligence caused by alleviating because of lead poisoning reduces, and has very much application prospect.
Biomaterial preservation
It is micro- to be preserved in China on November 29th, 2011 for a kind of lactobacillus plantarum (Lactobacillus plantarum)
Biological inoculum preservation administration committee common micro-organisms center, preservation address are in Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Institute of microbiology, the academy of sciences, state, deposit number are CGMCC 5494.
Detailed description of the invention
Fig. 1 is the dietary supplements production method process of probiotics and plant extracts；
Fig. 2 is protecting effect of the different protective agents to freeze-drying 5494 microbial activity of lactobacillus plantarum CGMCC；
Fig. 3 is protecting effect of the various concentration trehalose to freeze-drying 5494 microbial activity of lactobacillus plantarum CGMCC；
Fig. 4 is protecting effect of the various concentration skimmed milk to freeze-drying 5494 microbial activity of lactobacillus plantarum CGMCC；
Fig. 5 is that lactobacillus plantarum CGMCC 5494 is lyophilized in the dietary supplements tableting processes of probiotics and plant extracts
The influence of bacterium powder survival rate；
Fig. 6 is that lactobacillus plantarum CGMCC 5494 is lyophilized in the dietary supplements tableting processes of probiotics and plant extracts
The influence of bacterium powder viable count；
Fig. 7 is the dietary supplements procyanidins measurement standard curve of probiotics and plant extracts；
Fig. 8 is that epicatechin (EC), epigallocatechin (EGC), L-Epicatechin gallate (ECG) and table are not eaten
The HPLC-UV detection of sub- catechin and gallate (EGCG) standard substance；
Fig. 9 is the HPLC-UV detection of the dietary supplements of probiotics and plant extracts；
Figure 10 is mice organs, the variation of blood lead content；
Figure 11 is mouse liver, kidney, superoxide dismutase (SOD) variation of brain；
Figure 12 is mouse liver, catalase (CAT) variation of kidney；
Figure 13 is mouse liver, kidney, malonaldehyde (MDA) variation of brain；
Figure 14 is mouse liver, kidney, reduced glutathione (GSH) variation of brain；
Figure 15 is aminolevulinate dehydratase in mouse blood (ALAD) variation；
Figure 16 is zinc protoporphyrin in mouse blood (ZPP) variation；
Figure 17 is mouse step-through test index latency change；
Figure 18 is the variation of mouse step-through test index errors number.
Specific embodiment
Embodiment 1: influence of the different freezing drying protective agents in freezing dry process to lactobacillus plantarum survival rate
After lactobacillus plantarum CGMCC 5494 was activated for two generations in MRS fluid nutrient medium, 1L is accessed by 2% inoculum concentration
In MRS fluid nutrient medium, it is placed in constant temperature at 37 DEG C and expands culture 18h.5000g is centrifuged 15min at 4 DEG C, discards supernatant and collects bacterium
Body, thallus are washed twice with sterile phosphate buffer (PBS, 60mmol/L, pH6.5), collect thallus.Above-mentioned experiment repeats 5
It is secondary, 5 parts of thallus are collected, the sterile PBS solution of 25mL, 150g/L sucrose solution, 100g/L aqueous trehalose, 50g/ are separately added into
L sorbitol solution, 100g/L skimmed milk power solution are mixed as freezing drying protective agent and are resuspended.Protectant bacteria suspension will be contained
1mL is divided in sterile cillin bottle.Pre-freeze 3h at -80 DEG C.It is freeze-dried later, time 48h, vacuum degree 4Pa, cold-trap
- 50 DEG C of temperature.Freeze-drying front and back viable bacteria content is detected with bacterium colony colony counting method, number of viable is denoted as before being freeze-dried
M0；Number of viable is denoted as M after freeze-drying, obtains survival rate=M/M0× 100%.As a result as shown in Fig. 2:
As shown in Figure 2, the difference on effect of different freezing drying protective agents is obvious.10% trehalose, 10% degreasing
Milk solution effect is preferable, and survival rate is about 40% after freeze-drying.And sterile PBS solution protecting effect is worst, almost without work
With.The result shows that trehalose, skimmed milk reduce thallus in 5494 thallus freezing dry process of lactobacillus plantarum CGMCC
Cellular damage.Trehalose is selected, skimmed milk is convenient as the freezing drying protective agent ratio of lactobacillus plantarum CGMCC 5494.
The present invention considers the non-functional component for simplifying the dietary supplements of probiotics and plant extracts as far as possible, not right
The combination of a variety of freeze drying protectants is tested, and is only compared to the protecting effect of single kind freezing drying protective agent.
Embodiment 2: influence of the trehalose concentration to 5494 thallus freeze drying activity of lactobacillus plantarum CGMCC
The protective agent for preparing the various concentration gradient that trehalose concentration is 100g/L, 150g/L, 200g/L, 250g/L is molten
Liquid carries out expansion culture, centrifugation, washing and the freeze-drying of lactobacillus plantarum CGMCC 5494 according to the process of embodiment 1, living
Bacterium counts the concentration for investigating trehalose to the active influence of 5494 thallus of CGMCC freeze-drying.As a result as shown in Fig. 3:
As shown in Figure 3, with the raising of trehalose concentration, protecting effect is obviously improved.But experiment discovery, trehalose
When concentration is greater than 150g/L, the thallus viscosity after freeze-drying is larger, can not grind bacterium powder, this may be the water suction work of trehalose
Caused by, thus controlling trehalose concentration is that 100g/L is appropriate.
Embodiment 3: influence of the skimmed milk trehalose concentration to 5494 thallus freeze drying activity of lactobacillus plantarum CGMCC
The freeze-drying for preparing the various concentration gradient that skimmed milk concentration is 100g/L, 150g/L, 200g/L, 250g/L is protected
Agent solution is protected, expansion culture, centrifugation, washing and the freezing of lactobacillus plantarum CGMCC 5494 are carried out according to the process of embodiment 1
Dry, the concentration that count plate investigates skimmed milk is freeze-dried active influence to 5494 thallus of lactobacillus plantarum CGMCC.Knot
Fruit is as shown in Fig. 4:
As shown in Figure 4, with the raising of skimmed milk concentration, protecting effect is obviously improved.But skimmed milk concentration is more than
After 150g/L, influence of the concentration to protecting effect be not significant, 150g/L, 200g/L, the guarantor of tri- concentration skimmed milks of 250g/L
It is almost the same to protect effect, no significant difference (P > 0.05).After using each concentration degreasing milk solution as protective agent freeze-drying, thallus is ground
Bacterium powder is ground without influence.Comprehensively consider freezing dry process survival rate, the influence degree and tabletting that protective agent grinds bacterium powder
Situations such as bacterium powder activity change being likely to occur in journey, therefore select 100g/L aqueous trehalose and 150g/L degreasing milk solution
5494 freeze-dried vaccine powder of lactobacillus plantarum CGMCC is prepared for protective agent, further investigates tableting processes to probiotics and plant extract
The variation of microbial activity in the dietary supplements of object.
Embodiment 4: influence of the tableting pressure to the dietary supplements hardness of probiotics and plant extracts
The dietary supplements preparation process of probiotics and plant extracts is shown in attached drawing 1.
It is cold with 100g/L trehalose and 150g/L skimmed milk according to the component design of dietary supplements and the technique of setting
2 kinds of 5494 bacterium powder of freeze-drying lactobacillus plantarum CGMCC of dry protective agent preparation are main component, with plant extracts, auxiliary material to match
Grind method mixing.Distinguish tabletting with three pressure gear 1.2t, 2.4t, 3.6t pressure of tablet press machine, weight be 250mg ±
15mg.The dietary supplements made is detected into hardness (ordinary tablet 3-5kg) with hardness tester, and carries out high-altitude and fall experiment (taking
10 dietary supplements are placed in the vacancy 2M high, make its free-falling, if dietary supplements does not occur obvious slight crack or fracture fragmentation,
Then illustrate hardness qualification, every 3 repeated experiments), the results are shown in Table 2,
2 tableting pressure of table influences the dietary supplements hardness of probiotics and plant extracts
Note: different letters have significant difference (P < 0.05) between representing group
By table 2 it is found that after carrying out tabletting with 1.2t pressure, the dietary supplements hardness of two kinds of bacterium powder production is only respectively
2.31kg and 2.40kg, not up to 3-5kg standard.While high-altitude falls experiment and sliver phenomenon occurs, therefore 1.2t pressure is not
Preparation suitable for dietary supplements.And after carrying out tabletting with 3.6t pressure, the dietary supplements hardness of two kinds of bacterium powder production is
More than 6.10kg, and exceed national regulation 3-5kg standard.Therefore 3.6t pressure is unsuitable for the preparation of dietary supplements.And with
After 2.4t pressure carries out tabletting, the dietary supplements hardness of two kinds of bacterium powder production is respectively 4.20kg, 4.29kg, is met the requirements,
High-altitude, which falls to test, simultaneously does not occur sliver phenomenon.Therefore, 2.4t pressure is suitable for the preparation of dietary supplements, selects 2.4t pressure
Prepare dietary supplements.
Embodiment 5: influence of the tableting processes to 5494 microbial activity of lactobacillus plantarum CGMCC in dietary supplements
The dietary supplements tablet forming technique of probiotics and plant extracts is shown in attached drawing 1.
It, will be with according to the formula (table 1) of probiotics and the dietary supplements of plant extracts and the tablet forming technique of setting
100g/L trehalose and 150g/L skimmed milk are that 2 kinds of 5494 bacterium powder of freeze-drying lactobacillus plantarum CGMCC of cold dry protective agent preparation are
Main component, with plant extracts, auxiliary material with the mixing of facing-up method, by mixed powder through tablet press machine with 2.4t pressure tabletting, weight
For 250mg ± 15mg.Mixed powder is detected into number of viable with the method for plate culture count, is denoted as W0.With plate after tabletting
Colony counting method detects the number of viable in dietary supplements, is denoted as W.Then survival rate are as follows: survival rate=W/W0×
100%.As a result as shown in Figure 5, Figure 6:
Sheeting operation is larger to the activity and survival rate of different 5494 bacterium powder of lactobacillus plantarum CGMCC.By Fig. 5,
For Fig. 6 it is found that with 150g/L, 200g/L, 250g/L skimmed milk is that the bacterium powder of protective agent production is probiotics after main ingredient carries out tabletting
Damage it is larger, in the dietary supplements of probiotics and plant extracts 5494 survival rate of lactobacillus plantarum CGMCC less than 3%,
Survival volume is equal less than 2 × 109cfu/g.The bacterium powder made by protective agent of 10% trehalose is probiotics after main ingredient carries out tabletting
It can reach 18% or more with 5494 survival rate of lactobacillus plantarum CGMCC in the dietary supplements of plant extracts, survival volume is almost
It can reach 8 × 109cfu/g.Comparison result explanation, relative to skimmed milk, trehalose can be lactobacillus plantarum CGMCC 5494
More effectively protection is provided, damage of the CGMCC 5494 under the severe processing environment of high pressure is reduced, using trehalose as protective agent
5494 freeze-dried vaccine powder of CGMCC, be more suitable for the production of the dietary supplements of probiotics and plant extracts.
Embodiment 6: influence of the tableting processes to the dietary supplements procyanidins of probiotics and plant extracts
Procyanidine is a kind of plant flavone, usually grape seed extract or French maritime pine bark extract.It is former
Anthocyanidin is a kind of novel high-efficiency antioxidant agent, is one of free radical scavenger most potent at present.Measuring method is molysite
Catalytic colorimetry measures mixed powder and dietary supplements procyanidins content after tabletting before tabletting.It is specific as follows:
The preparation of 20% ammonium ferric sulfate solution: weighing 10.0g ammonium ferric sulfate, is settled to 50mL with 2mol/L HCl dissolution.
The preparation of reaction mixture: taking n-butanol, concentrated hydrochloric acid, 20% ammonium ferric sulfate, 85:5:0.4 is prepared by volume.
Sample treatment: dietary supplements is in 10ml brown capacity after accurately weighing before 140mg tabletting mixed powder or tabletting
In bottle, dehydrated alcohol dissolution is settled to scale.Supersonic oscillations extract 10min, 2-3 times repeatedly, shake up.Take supernatant mistake
0.22 μm of film to get arrive sample liquid.
The preparation of blank sample: directly dehydrated alcohol is taken to be settled to scale.Supersonic oscillations extract 10min, repeatedly 2-3
It is secondary, it shakes up.Supernatant is taken to cross 0.22 μm of film to get sample liquid is arrived.
The drafting of standard curve: procyanidine standard items 5mg is accurately weighed in 10ml brown volumetric flask, uses dehydrated alcohol
It dissolves, be settled to scale, the titer that concentration is 0.50mg/mL is made.Take the titer dehydrated alcohol of appropriate 0.50mg/mL
Dilution is made the standard series that concentration is 0,10,25,50,100,150,200,250,300 μ g/mL and uses liquid.It draws respectively each
Concentration in 10mL scale test tube, is added 9.0mL reaction mixture, stoppers plug, shake up, be placed in boiling water bath using liquid 1.0mL
After middle heating 40min, the cooling 4min-5min of ice water of taking-up immediately takes out, restores to room temperature, n-butanol is settled to scale
Line stoppers plug, shakes up, and is returned to zero at 550nm with reagent blank, draws light absorption value-procyanidin concentration standard curve.Mark
Directrix curve is shown in attached drawing 7.
The measurement of procyanidin content: taking sample liquid 1.0mL in 10mL scale test tube, and 9.0mL reaction mixture is added,
Plug is stoppered, is shaken up, is placed in boiling water bath after heating 40min, the cooling 4min-5min of ice water of taking-up immediately takes out, restores extremely
After room temperature, n-butanol is settled to graduation mark, stoppers plug, shakes up, and is returned to zero at 550nm with reagent blank, measurement absorbance with
Standard curve comparison, finds procyanidin content, the results are shown in Table 3:
Influence of the dietary supplements tableting processes of 3 probiotics of table and plant extracts to procyanidine
By table 3 it is found that mixed powder is by molysite catalytic colorimetry detected level up to 29.17mg/g, tabletting before tabletting
Afterwards in dietary supplements detected level up to 29.52mg/g.Recall rate is up to 101.20% before and after tabletting.Illustrate tableting processes for
Procyanidine has little effect.
Embodiment 7: influence of the tableting processes to tea polyphenols in the dietary supplements of probiotics and plant extracts
The general name of Polyphenols of Tea is known as tea polyphenols.Catechin compounds are the bulk compositions of tea polyphenols, main
It to include epicatechin (EC), epigallocatechin (EGC), L-Epicatechin gallate (ECG) and epi-nutgall catechu
Plain 4 kinds of substances of gallate (EGCG).With meal supplement after mixed powder before high effective liquid chromatography for measuring tabletting and tabletting
EC in agent, EGC, ECG, EGCG content.It is specific as follows:
The processing of standard items: mixed mark production accurately weighs EC, EGC, ECG, and each 10mg of EGCG standard items is placed in 100mL capacity
Bottle, distilled water are settled to scale and obtain hybrid standard product solution.Single mark production accurately weighs EC standard items 10mg merging 100mL and holds
Measuring bottle, distilled water are settled to scale and obtain EC standard solution.EGC, the mono- mark of ECG, EGCG are made again according to this step.
The processing of sample: dietary supplements and 2.0mL centrifuge tube after mixed powder or tabletting are accurately weighed before 50mg tabletting
In, 70 DEG C of 1mL 50% preheated methanol solutions are added, mix.It moves into 70 DEG C of water-baths immediately, extracts 20min, every
5min stirring is primary.10min is centrifuged with 3500r/min after being cooled to room temperature, takes supernatant to cross 0.22 μm of film, obtains sample treatment
Liquid is spare.
Agilent quaternary pump chromatograph, Agilent C18 (4.6mm × 250mm, 5 μm) chromatographic column, chromatographic condition are as follows:
Mobile phase A phase: water (contains 0.05% trifluoroacetic acid).
Mobile phase B phase: acetonitrile.
Gradient condition:
1. starting 90%A phase, 10%B phase
2. 90%A phase in 20min, 10%B phase → 70%A phase, 30%B phase
3. 70%A phase, 30%B phase keeps 4min
4. 70%A phase in 1min, 30%B phase → 90%A phase, 10%B phase
5. 90%A phase keeps 5min
Sample volume: 5 μ L
Flow velocity: 1mL/min
Column temperature: 30 DEG C
Detector: UV detector, Detection wavelength: 280nm
Measurement list mark, mixes mark, and, peak area quantification qualitative with retention time measures mixed powder and meals after tabletting before tabletting
Eat EC, EGC, ECG in replenishers, EGCG content.Standard substance map is shown in Fig. 8, and powder and viable bacteria tablet map example are shown in Fig. 9.
Experimental result is as shown in table 4:
Influence of the 4 dietary supplements tableting processes of table to tea polyphenols main component
By table 4 it is found that mixed powder EGC, EC, EGCG, ECG detected level are respectively 2.59mg/g, 1.24mg/ before tabletting
G, 7.68mg/g, 1.53mg/g.EGC, EC, EGCG, ECG detection in the dietary supplements of probiotics and plant extracts after tabletting
Amount is respectively 2.61mg/g, 1.23mg/g, 7.59mg/g, 1.53mg/g.Recall rate is respectively up to 101.20% before and after tabletting,
99.19%, 98.28%, 100.00%.Illustrate that tableting processes have little effect tea polyphenols main component.
Embodiment 8: storage requirement is to the active shadow of lactobacillus plantarum in the dietary supplements of probiotics and plant extracts
It rings
By the dietary supplements of the probiotics and plant extracts that are made in embodiment 5 (using 10% trehalose as protective agent)
With aluminium foil bag vacuum heat-seal packaging, be protected from light, keep away it is wet, keep away oxygen, stored under the conditions of being placed in 4 DEG C and 20 DEG C, measurement store 15 days, 30
It, 90 days, 5494 number of viable of lactobacillus plantarum CGMCC after 180 days, investigate storage temperature and period of storage to probiotics
With the active influence of dietary supplements of plant extracts.With the meals of the method for plate culture count detection probiotics and plant extracts
Viable count of the replenishers when storing initial is eaten, C is denoted as0.Storage measures probiotics after a certain period of time, with the method for plate culture count
With the viable count of the dietary supplements of plant extracts, it is denoted as C, then stores survival rate are as follows: storage survival rate=C/C0×
100%.As a result such as table 5, shown in table 6:
5 dietary supplements of table, 5494 activity change of lactobacillus plantarum CGMCC in storage process under the conditions of 4 DEG C
6 dietary supplements of table, 5494 activity change of lactobacillus plantarum CGMCC in storage process at 20 °C
By table 5, table 6 is stored, probiotics and plant extracts under the conditions of 4 DEG C it is found that store under the conditions of compared to 20 DEG C
Dietary supplements activity decline it is slower.5494 viable count of lactobacillus plantarum CGMCC is still greater than after storage 1 month
108CFU/g, viable count is still greater than 10 after storage 6 months5CFU/g.And meals of probiotics and plant extracts at 20 °C
The activity for eating replenishers declines faster, and 5494 viable count of lactobacillus plantarum CGMCC only has 10 after storage 1 month5CFU/g, storage
After 6 months viable count only residue 103CFU/g illustrates that low-temperature storage is more advantageous to the diet for keeping probiotics and plant extracts
The activity and extension shelf life of replenishers.
9 period of storage of embodiment, storage temperature is to the dietary supplements procyanidins of probiotics and plant extracts
Influence experiment
By the dietary supplements of the probiotics and plant extracts that are made in embodiment 5 (using 10% trehalose as protective agent)
With aluminium foil bag vacuum heat-seal packaging, be protected from light, keep away it is wet, keep away oxygen, stored under the conditions of being placed in 4 DEG C and 20 DEG C.According to the side in embodiment 5
The dietary supplements procyanidins of method, probiotics and plant extracts after measurement storage 15 days, 30 days, 90 days, 180 days contain
Amount.Investigate the influence of storage temperature and period of storage to the dietary supplements procyanidins of probiotics and plant extracts.Knot
Fruit is as shown in table 7:
7 dietary supplements of table storage process procyanidins changes of contents under the conditions of 4 DEG C, 20 DEG C
Note: each group is without significant difference
By table 7 it is found that after storage 15 days between each group probiotics and plant extracts dietary supplements procyanidins
There is decline compared with when storing initial but difference is not significant, but as the content of the extension procyanidine of period of storage is not significant
Difference illustrates that procyanidine is not degraded during storage 15 to 180 days.Although illustrating the packaging energy of aluminium foil bag vacuum heat-seal
Enough starvations, but a little degradation that a little oxygen causes procyanidine may have been remained in packaging process.But aluminium foil bag is true
The packaging of sky heat-sealing, is protected from light, and keeps away wet, and the preservation condition for keeping away oxygen prevents procyanidine to continue to degrade, and illustrates aluminium foil bag vacuum
The packaging of heat-sealing is protected from light, keeps away preservation condition that is wet, keeping away oxygen and be conducive to protect in the dietary supplements of probiotics and plant extracts
Antioxidant, extend shelf life.
Embodiment 10: influence of the storage requirement to tea polyphenols in the dietary supplements of probiotics and plant extracts
By the dietary supplements of the probiotics and plant extracts that are made in embodiment 5 (using 10% trehalose as protective agent)
With aluminium foil bag vacuum heat-seal packaging, be protected from light, keep away it is wet, keep away oxygen, stored under the conditions of being placed in 4 DEG C and 20 DEG C.According to the side in embodiment 6
Method measures tea polyphenols in the dietary supplements of the probiotics and plant extracts after storing 15 days, 30 days, 90 days, 180 days and contains
Amount.Investigate the influence of storage temperature and period of storage to tea polyphenols in the dietary supplements of probiotics and plant extracts.As a result
Such as table 8, table 9, table 10, shown in table 11:
8 dietary supplements of table epigallocatechin changes of contents in storage process under the conditions of 4 DEG C, 20 DEG C
Note: different letters have significant difference (P < 0.05) between representing group
9 dietary supplements of table epicatechin (EC) changes of contents in storage process under the conditions of 4 DEG C, 20 DEG C
Note: each group is without significant difference
10 dietary supplements of table Epigallo-catechin gallate (EGCG) content in storage process under the conditions of 4 DEG C, 20 DEG C
Variation
Note: different letters have significant difference (P < 0.05) between representing group
L-Epicatechin gallate (ECG) contains quantitative change to 11 dietary supplements of table in storage process under the conditions of 4 DEG C, 20 DEG C
Change
Note: different letters have significant difference (P < 0.05) between representing group
By table 8, table 9, table 10, table 11 is it is found that the diet of probiotics and plant extracts is mended between each group after storage 15 days
The content for filling EGC, ECG and EGCG in agent is decreased significantly compared with when storing initial, and EC has decline compared with when storing initial
But difference is not significant, but as the content of the extension EC, EGC, ECG, EGCG of period of storage are not significantly different, illustrates EC,
EGC, ECG, EGCG do not degrade during storage 15 to 180 days.Although illustrating that the packaging of aluminium foil bag vacuum heat-seal can be every
Anoxybiotic gas, but a little oxygen may have been remained in packaging process and caused EC, EGC, ECG, a little degradation of EGCG.But aluminium foil bag
The packaging of vacuum heat-seal, is protected from light, and keeps away wet, and the preservation condition for keeping away oxygen prevents EC, EGC, ECG, and EGCG's continues to degrade, and illustrates aluminium
The packaging of foil bag vacuum heat-seal is protected from light, keeps away the diet that preservation condition that is wet, keeping away oxygen is conducive to protect probiotics and plant extracts
Tea polyphenols main component in replenishers, extends shelf life.
Embodiment 11: dietary supplements acts on the intervention reduction of lead content in lead exposure Mice Body
Health male C57BL/6 mouse 40 for taking weight 20-25g, is randomly divided into 4 groups: blank control group, lead exposure mould
Dietary supplements intervention group, the DMSA group of type group, probiotics and plant extracts, every group of 10 mouse.Experiment 61 days by a definite date, it is empty
Normal water is given in 56 days sterile PBS solutions of daily stomach-filling 0.3mL before white control group, last not stomach-filling in five days.Lead exposure model
The first 56 days sterile PBS solutions of daily stomach-filling 0.3mL of group give leaded drinking-water and (lead acetate are dissolved in drinking water, makes its concentration
For 1g Pb/L, freely drunk for mouse), normal water is given in last not stomach-filling in five days.The meals of probiotics and plant extracts
Meal supplement agent solution (the Example 5 of 56 days daily stomach-filling 0.3mL probiotics and plant extracts before food replenishers intervention group
In the dietary supplements of a piece of probiotics and plant extracts that is prepared using 10% trehalose as protective agent to be dissolved in 0.6mL sterile
In PBS solution), leaded drinking-water is given, normal water is given in last not stomach-filling in five days.DMSA group first 56 days daily stomach-filling 0.3mL
Sterile PBS solution, gives leaded drinking-water, and the DMSA solution of last 5 days daily stomach-filling 0.3mL 5mg/mL gives normal water.
Mouse is put to death after experiment, takes blood, liver, kidney, brain.Precision weighs blood, liver, kidney, brain weight, after pure nitric acid is added
It is put into Hyperfrequency waves eliminating stove and clears up, time 20min.Obtained digestion solution is contained using atomic absorption spectroscopy determination lead
Amount.As a result as shown in Fig. 10:
As shown in Figure 10, blank control group, lead exposure model group, the meal supplement of probiotics and plant extracts are compared
The liver of agent intervention group, DMSA (traditional heavy metal detoxification agent) group, kidney, brain, blood lead content is it is found that probiotics and plant mention
Take the dietary supplements of object, DMSA can significantly reduce mouse liver, kidney, brain, lead content in blood.Illustrate probiotics and
The dietary supplements of plant extracts has the function of reducing lead content in Mice Body.
Embodiment 12: dietary supplements alleviates the intervention of lead exposure oxidative damage
Zoopery is carried out by grouping described in embodiment 11, mouse is put to death after experiment, takes blood, liver, kidney,
Brain.Superoxide dismutase (SOD), catalase are measured according to the method that Bioengineering Research Institute's kit is built up in Nanjing
(CAT), reductive glutathione (GSH), glutathione peroxidase (GSH-Px), malonaldehyde (MDA) content or level.
Test result such as attached drawing 11, attached drawing 12, attached drawing 13, shown in attached drawing 14:
Superoxide dismutase (SOD), catalase (CAT) are antioxidases important in body, and oxidation is answered
Swash one of damage markers.Reduced glutathione (GSH) is the important antioxidant of the intracorporal one kind of biology.Malonaldehyde
It (MDA) is a kind of lipid peroxide, content can reflect the level of lipid peroxidation and cellular damage degree of body.Pass through
It is found that compared to blank control group, the activity of the SOD of lead exposure model group, CAT are significantly reduced by Figure 11, Figure 12, Figure 13, Figure 14,
GSH content is remarkably decreased, and MDA content significantly rises.And lead exposure model group is compared, the diet of probiotics and plant extracts is mended
It filling agent intervention group and has significantly restored liver, kidney, SOD activity caused by lead exposure reduces in brain tissue, significantly restore liver,
The activity of CAT reduces in renal tissue, significantly improves liver, kidney, and the GSH content in brain tissue reduces MDA content.
Succimer (DMSA) is a quasi-tradition heavy metal competitiveness antidote, comparison DMSA group discovery, probiotics and plant extract
The dietary supplements of object effect ratio DMSA in terms of alleviating oxidativestress damage caused by lead exposure is stronger.In summary as a result,
Illustrate that the dietary supplements of probiotics and plant extracts effectively alleviates oxidativestress damage caused by lead exposure.
Embodiment 13: dietary supplements, which alleviates aminolevulinate dehydratase activity in lead exposure Mice Body, reduces experiment
Zoopery is carried out by grouping described in embodiment 11, mouse is put to death after experiment, takes blood, 4 DEG C of standing 2h
3000r/min is centrifuged 10min afterwards, takes upper serum.It is measured according to the method for Shanghai Chuan Xiang Biotechnology Co., Ltd kit
Aminolevulinate dehydratase (ALAD) activity, test result are as shown in Fig. 15:
Aminolevulinate dehydratase directly participates in the biosynthesis of body ferroheme, with the direct phase of body hemopoietic system
It closes.As shown in Figure 15, blank control group is compared, the activity of lead exposure model group ALAD significantly reduces.And probiotics and plant
The dietary supplements intervention group of extract has significantly restored the activity of ALAD, and effect is suitable with tradition heavy metal detoxification agent DMSA.
Illustrate that the dietary supplements of probiotics and plant extracts effectively alleviates aminolevulinate dehydratase caused by lead exposure
(ALAD) Active lesions.
Embodiment 14: dietary supplements alleviates zinc protoporphyrin content in lead exposure Mice Body and rises experiment
Zoopery is carried out by grouping described in embodiment 11, mouse is put to death after experiment, takes blood, 4 DEG C of standing 2h
3000r/min is centrifuged 10min afterwards, takes upper serum.It is measured according to the method for Shanghai Chuan Xiang Biotechnology Co., Ltd kit
Zinc protoporphyrin (ZPP) content, test result are as shown in table 20:
The content of zinc protoporphyrin (ZPP) is one of index of lead poisoning in blood.By table 20 it is found that comparing blank control
Group, lead exposure model group ZPP content significantly rise.And the dietary supplements intervention group of probiotics and plant extracts significantly reduces
ZPP content, effect are suitable with tradition heavy metal detoxification agent DMSA.Illustrate that the dietary supplements of probiotics and plant extracts has
The content for alleviating zinc protoporphyrin caused by lead exposure (ZPP) of effect rises, and alleviates lead poisoning.
Embodiment 15: dietary supplements alleviates lead exposure intelligent of ageing mice injury experiment
Step-through test is utilized mouse and becomes dark habit, keeps away concealed installation and sets and is divided into light room and darkroom.Light room illumination is 25W, darkroom
It is not provided with illumination, there is the connection of the channel 3cm between light room darkroom, it can random open and close.Light room, darkroom bottom are paved with copper grid, secretly
Room base copper grid can be powered, voltage 24V.Mouse enters darkroom and is shocked by electricity.Experiment is divided into training, formally tests two parts.
Training makes mouse possess electric shock memory, and end is formally tested afterwards for 24 hours, formal experimental observation mouse incubation period (mouse first
The secondary time completely into darkroom), into the errors number in darkroom, investigate the ability of learning and memory of mouse.The specific method is as follows:
Zoopery is carried out by grouping described in embodiment 11, mouse step-through test is carried out after stomach-filling.Step-through test first day
Morning 10:00 starts to train.Training time is 3min, and all mouse are individually placed among bright room, body back to channel after starting
Body not face hidden door, channel is closed between light room darkroom.Channel between light room darkroom is opened after adapting to 1min, starts simultaneously at meter
When.Mouse enters behind darkroom shocked by electricity after take out immediately.The next morning 10:00 starts formally to test, formal experimental period
For 5min, electric current is not added in darkroom.All mouse are individually placed among bright room back to channel after experiment starts, body not face
Hidden door, channel is closed between light room darkroom.Channel between light room darkroom is opened after adapting to 1min, starts simultaneously at timing.Record
Mouse incubation period and the errors number into darkroom in 5min.Experimental result such as attached drawing 17, shown in attached drawing 18:
By Figure 17, Figure 18 is it is found that compare blank control group, and lead exposure model group incubation period is obviously shortened, errors number
It is obvious to rise.The ability of learning and memory of mouse receives damage.And the dietary supplements intervention group of probiotics and plant extracts
Significantly shorter latencies are restored, the intellectual damage that errors number rises, effect is more stronger than traditional heavy metal detoxification agent DMSA.It says
The dietary supplements of bright probiotics and plant extracts has effectively restored the ability of learning and memory of mouse.
Embodiment 16: preparing according to the following ratio while the dietary supplements containing profitable probliotics and plant extracts
(1) the lactobacillus plantarum CGMCC after spreading cultivation and washing is resuspended using 10% trehalose as freeze drying protectant
5494, make viable bacteria concentration up to 1011CFU/mL or more is freeze-dried after preculture 60min at 37 DEG C of temperature and is lyophilized
Bacterium powder.
(2) according to parts by weight, by 37.9% microcrystalline cellulose, 4% low-substituted hydroxypropyl cellulose, 0.1%
Superfine silica gel powder, 5% grape seed extract, 3% tea polyphenols, 50% bacterium powder, mixing, with facing-up method mix, obtain powder at
Product.
(3) directly use tablet press machine with 2.4t pressure tabletting powder obtained in the previous step, obtained tablet weight is 250mg
±15mg。
The dietary supplements active component content obtained by this method is high, shelf life is long；Prebiotic bacterial content can reach 8.3 ×
109cfu/g, viable count is still greater than 10 after storing 6 months at 4 DEG C6CFU/g；Anthocyanidin content reach 29.17mg/g, EGC, EC,
The content of EGCG, ECG are respectively up to 2.61mg/g, 1.23mg/g, 7.59mg/g, 1.53mg/g, after storing 6 months at 20 DEG C still
There is 90% or more retention rate；The lead of the lead content and blood that can reduce the internal organs such as liver in Mice Body, kidney, brain simultaneously contains
Amount alleviates oxidativestress damage caused by the lead poisoning of Mice Body internal cause, aminolevulinate dehydratase activity reduces, zinc protoporphyrin
Content rises, and intelligence caused by alleviating because of lead poisoning reduces.
Embodiment 17 prepares according to the following ratio while the dietary supplements containing profitable probliotics and plant extracts
(1) the lactobacillus plantarum CGMCC after spreading cultivation and washing is resuspended using 10% trehalose as freeze drying protectant
5494, make viable bacteria concentration up to 1011CFU/mL or more is freeze-dried after preculture 60min at 37 DEG C of temperature and is lyophilized
Bacterium powder.
(2) according to parts by weight, by 59.7% microcrystalline cellulose, 6% low-substituted hydroxypropyl cellulose, 0.3%
Superfine silica gel powder, 2.5% grape seed extract, 1.5% tea polyphenols, 30% bacterium powder), mixing is mixed with facing-up method, obtains powder
Expect finished product.
(3) directly use tablet press machine with 2.4t pressure tabletting powder obtained in the previous step, obtained tablet weight is 250mg
±15mg。
The dietary supplements active component content obtained by this method is high, shelf life is long；Prebiotic bacterial content can reach 8.3 ×
109cfu/g, viable count is still greater than 10 after storing 6 months at 4 DEG C6CFU/g；Anthocyanidin content reach 29.17mg/g, EGC, EC,
The content of EGCG, ECG are respectively up to 2.61mg/g, 1.23mg/g, 7.59mg/g, 1.53mg/g, after storing 6 months at 20 DEG C still
There is 90% or more retention rate；The lead of the lead content and blood that can reduce the internal organs such as liver in Mice Body, kidney, brain simultaneously contains
Amount alleviates oxidativestress damage caused by the lead poisoning of Mice Body internal cause, aminolevulinate dehydratase activity reduces, zinc protoporphyrin
Content rises, and intelligence caused by alleviating because of lead poisoning reduces.
Although the present invention has been described by way of example and in terms of the preferred embodiments, it is not intended to limit the invention, any to be familiar with this skill
The people of art can do various change and modification, therefore protection model of the invention without departing from the spirit and scope of the present invention
Enclosing subject to the definition of the claims.
Claims (4)
1. a kind of dietary supplements containing profitable probliotics and plant extracts simultaneously, which is characterized in that the dietary supplements is
To be freeze-dried the probiotics bacterial powder obtained as main ingredient, it is aided with grape seed extract, adds after proper auxiliary materials with powder through tabletting
What method was prepared, according to parts by weight containing the microcrystalline cellulose of 37.9-59.7%, the low substituted hydroxy-propyl fiber of 4-6%
Element, the superfine silica gel powder of 0.1-0.3%, the grape seed extract of 2.5-5%, the tea polyphenols of 1.5-3%, the bacterium powder of 30-50%；Institute
Stating probiotics is the lactobacillus plantarum (Lactobacillus plantarum) that deposit number is CGMCC 5494, in 2011
On November 29, in is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center；It is described freeze-drying be with
The trehalose of 100-150g/L or the degreasing milk solution of 100-250g/L are as freeze drying protectant；It is planted in the dietary supplements
The viable bacteria content of object lactobacillus CGMCC 5494 reaches 109CFU/g or more；The pressed disc method carry out tabletting be through tablet press machine with
2.4t pressure tabletting, obtained tablet weight are 250mg ± 15mg.
2. dietary supplements according to claim 1, which is characterized in that the freeze-drying is the trehalose with 100g/L
As freeze drying protectant.
3. dietary supplements according to claim 1, the probiotics bacterial powder the preparation method is as follows: by lactobacillus plantarum
CGMCC 5494 expands culture, centrifugation, washing after activating, freeze drying protectant is then added, thallus is resuspended, reach cell concentration
1011CFU/mL or more, pre-freeze after mixing, is then freeze-dried.
4. dietary supplements according to claim 1, the dietary supplements, according to parts by weight containing 48.9% it is micro-
Crystalline cellulose, 5% low-substituted hydroxypropyl cellulose, 0.1% superfine silica gel powder, 3.6% grape seed extract, 2.4%
Tea polyphenols, 40% bacterium powder.
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