CN109125180A - A kind of preparation and its detection test method of Traditional Chinese medicinal hand lotion - Google Patents
A kind of preparation and its detection test method of Traditional Chinese medicinal hand lotion Download PDFInfo
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- CN109125180A CN109125180A CN201811196534.0A CN201811196534A CN109125180A CN 109125180 A CN109125180 A CN 109125180A CN 201811196534 A CN201811196534 A CN 201811196534A CN 109125180 A CN109125180 A CN 109125180A
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
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- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
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Abstract
Traditional Chinese medicinal hand lotion bactericidal effect of the invention is good, the detection test method of Traditional Chinese medicinal hand lotion can the bacteriostasis rate to Traditional Chinese medicinal hand lotion accurately calculated, clearly define the bactericidal effect of effective Traditional Chinese medicinal hand lotion and for the bacteria-eliminating efficacy of Traditional Chinese medicinal hand lotion detection determination provide practicable detection test method.The present invention is by the setting of pallet monomer so that the filling convenience and high-efficiency of bacteriostatic hand sanitizer, filling in the intracorporal bacteriostatic hand sanitizer of hand cleanser chamber is 1-2ml, both can satisfy and disposable when washing one's hands needed in turn avoid conventional hand cleanser bring bacterial cross-infection is used for multiple times.
Description
Technical field
The invention belongs to hand cleanser fields, preparation and its detection test method more particularly to Traditional Chinese medicinal hand lotion.
Background technique
Hand cleanser be it is a kind of cleaning hand based on skin care cleaning solution, in the modern life sterilizing be wash one's hands it is main
Purpose, and hand cleanser can only meet the effect of its sterilization, because all containing alcohol in most of hand cleanser, alcohol cannot be effectively
The some dirts being attached in skin fine gap, such as dust, soil, bloodstain are removed, so, once it is infected with such dirt on hand
Object is only inadequate with hand cleanser, so cannot replace soap in some necessary occasion hand cleanser.As it can be seen that using hand cleanser
On condition that first these dirts are got rid of.Therefore, it should be washed one's hands in this case using soap, and should not be merely using washing one's hands
Liquid.In fact, go to hospital it finds that, many doctors use washing hand with soap, and reason is not due to the cheap of soap.
Hand cleanser can only be the dirt that first washes one's hands with soap as the reserves of soap, best hand washing, reuse hand cleanser
Sterilizing skin moistening.It is also not the more the better when with hand cleanser, a drop is pressed out when using every time and is sufficient, and is being rubbed with the hands repeatedly on hand
After washing, then with circulating water flushing 15 seconds or more, thus hand cleanser can be thoroughly cleaned up.
Application No. is the applications of CN201710458584.0 to disclose a kind of flushing-free bacteriostatic hand sanitizer and preparation method thereof,
It has obtained Hand-washing Solution Exempt From Rinse according to certain sequence mixed configuration by certain component, has obtained degerming effect by centainly testing
The preferable scheme of fruit, convenient to carry out and flushing-free are easy to use；But since it does not carry out the packaged form of hand cleanser
Change, easy cross-infection when there are problems that more people use using traditional wheel cylinder formula usage mode.
Application No. is the application documents of CN201310580014.0 to disclose a kind of degerming Environmentally-friehand hand sanitizer, and hand cleanser is not only
Also there is sterilizing function with dirt-removing functions；But due to its hand cleanser preparation process be only by grape seed extract, hexadecanol,
Soap lye etc. directly mixes and does not specify its extraction process, implements more difficulty, while also not disclosing its specific sterilization test
And test method, therefore its bactericidal effect and unreliable.
Current hand cleanser on the market is relatively fixed in preparation method, and constituent is mostly chemical products, cannot be made
Lesser natural traditional Chinese medicine ingredient preparation is injured to human hand with naturally no added；And the bactericidal effect of Traditional Chinese medicinal hand lotion is detected
Test method majority is obtained by true man's Experimental comparison's effect, and this test method is not only time-consuming but also effect is also due to people
It is different.
Summary of the invention
The present invention overcomes disadvantages described above to provide the preparation and its detection test method of a kind of Traditional Chinese medicinal hand lotion, manufactured Chinese medicine
Hand cleanser products integrally have preferable disinfection effect, and the detection test method of Traditional Chinese medicinal hand lotion can be to Traditional Chinese medicinal hand lotion
Bacteriostasis rate accurately calculated, clearly define the bactericidal effect of effective Traditional Chinese medicinal hand lotion and for Traditional Chinese medicinal hand lotion degerming imitate
Fruit detection determination provides practicable detection test method.
The preparation of hand cleanser: extracting honeysuckle, Fructus Forsythiae, mulberry leaf, peppermint crush, and cross No. 3 sieves；Accurately weighed honeysuckle 0.5g,
Fructus Forsythiae 0.5g, mulberry leaf 0.1g, peppermint 0.5g, mixing are added 50% ethyl alcohol 50mL reflux 1h and obtain extracting solution, and extracting solution is cooled to
Room temperature, 4000rpm are centrifuged 10min, filtering, and dehydrated alcohol 100mL, ultrasonic 30min, supersonic frequency is added to doing in recycling design
40kHz, ultrasonic power 400W, filtering.14mL filtrate is taken, 14mL dehydrated alcohol is added, 12mL water mixes, molten according to every 10mL
25mg chlorhexidine acetate is added in liquid, and the ratio of 125 μ L glucose acid Chlorhexidines prepares hand cleanser sample to be tested, mixes to get this
The bacteriostatic hand sanitizer of invention.
Wherein honeysuckle, Fructus Forsythiae, mulberry leaf, peppermint carry out cleaning first spare respectively, lyophilization are carried out to Fructus Forsythiae, to gold
Honeysuckle flower, mulberry leaf, peppermint carry out hot drying, are disposed after above-mentioned raw material, it is passed through to multistage plant ultra-micropowder grinding respectively
Ultramicro-powder is made in technology, and by the ultra micro amylaceous substance of ultramicrotechnique production after pulverizing, the powder of formation is provided with good
Good dispersibility and resolvability provides the foundation for good dissolution mixing.
Lyophilization is carried out to Fructus Forsythiae, after carrying out hot drying to honeysuckle, mulberry leaf, peppermint, obtained lyophilization is connected
It sticks up and Ultramicro-powder is made by multistage plant ultra-micropowder grinding technology respectively in heat drying honeysuckle, mulberry leaf, peppermint.To gold and silver
The drying temperature that flower, mulberry leaf, peppermint carry out hot drying is 120 DEG C or so.Lyophilization is carried out to Fructus Forsythiae using freeze drier.
In above-mentioned preparation method, the process that extracting solution is cooled to room temperature carries out in two stages, and the first stage is that water-bath is protected
Thermophase continues 1h or so, guarantee honeysuckle, Fructus Forsythiae, mulberry leaf, peppermint ultra micro grinding can be sufficiently mixed and be substantially dissolved in
In extracting solution；Second stage is the rapid cooling stage, guarantees to be rapidly cooled to room temperature in 3min, to further increase hand cleanser
The content of effective traditional Chinese medicine ingredients of middle dissolution.
The present invention carries out bacteriostatic test using following test method:
Step 1: by instrument apparatus high pressure sterilization, taking three test tubes, mark Candida albicans, Escherichia coli, golden yellow respectively
Color staphylococcus.In the test tube of label Candida albicans, SDB culture medium 5mL and Candida albicans 5mL is added；It is big in label
In the test tube of enterobacteria, TSB culture medium 5mL and Escherichia coli 0.5mg is added；In the test tube of label staphylococcus aureus,
TSB 5mL and staphylococcus aureus 0.5mL is added.
Step 2: taking two test tubes, mark TSB, SDB as reference substance respectively.TSB is added in the test tube of label TSB
5mL is added SDB 5mL in the test tube of label SDB and five test tubes is placed in constant incubator.After viable bacteria, PBS is used
1000 times of (phosphate buffer solution) dilution, it is spare.
Step 3: the small filter paper of 1 × 1cm of preparation is several, is respectively put into label Candida albicans, Escherichia coli, golden yellow
In staphylococcic culture dish, it is spare that corresponding diluted strain 20 μ L, 37 DEG C of drying 30min is added dropwise in each small filter paper.
Step 4: 32 culture dishes being taken to mark Escherichia coli, staphylococcus aureus, Candida albicans, Escherichia coli respectively
+ medical fluid, staphylococcus aureus+medical fluid, Candida albicans+medical fluid 2min, 5min, 10min, 20min, control group TSA (Sharpe
Agar medium) and SDA (soybean casein agar) 2min, 5min, 10min, 20min respectively, it is added in culture medium and prepares
TSA or SDA, rotate culture dish, keep its full and uniform, place solidification.
Step 5: the scraps of paper of drying being put into the cillin bottle containing medical fluid or PBS (phosphate buffered saline solution), start to count
When, respectively when acting on 2min, 5min, 10min, 20min, print is put into the cillin bottle containing 5mL PBS with sterile tweezer
In, it mixes, draws 1mL respectively in the corresponding culture dish solidified, smoothened with spreading rod, culture is overturn after agar solidification
Ware is set in constant incubator, and the counting of viable bacteria bacterium colony is made.
Step 6: carrying out experimental result calculating；Candida albicans culture 48h, Escherichia coli, staphylococcus aureus culture
After for 24 hours, culture dish is taken out, by formula: X=(A-B)/A × 100%；Wherein X indicates bacteriostasis rate (%)；A indicates that control sample is flat
Equal clump count；B indicates subject sample average clump count.
Including pallet monomer, the groove and convex block of tray surface is arranged in Traditional Chinese medicinal hand lotion production equipment of the invention；
The pallet monomer be can vacuum-packed enclosed construction, bottom is provided with several vacuum suction holes, and vacuum suction hole can be right
It is vacuumized inside pallet monomer；The upper surface of pallet monomer is placed with film, and film is laid on pallet monomer by convex block support；
When the vacuum suction hole vacuumizes, the film formed on pallet monomer it is wavy, wavy film upper surface have washes
Hand sap cavity body.
The Traditional Chinese medicinal hand lotion production equipment further includes feeder pipe, and feeder pipe is needle-shaped injection device, lifting rate size
For every drop 1ml or so.
Since above-mentioned bacteriostatic hand sanitizer of the invention is potent antibacterial hand cleanser, every time when filling bacteriostatic hand sanitizer only
Feeder pipe blanking 1 to 2 is needed to drip；The volume of the hand cleanser cavity is 2ml.
The hand cleanser cavity is that substantially semi-cylindrical, pallet monomer upper surface convex block are triangular bump, each
The top of triangular bump is on sustained height, after filling hand cleanser is in hand cleanser cavity, water soluble film is covered on thin
Above film, seal to form several hand cleanser monomers in its contact point using heat sealing machine.
Soap layer may further be coated on the hand cleanser monomer, the thickness of soap layer is preferably 200 μm.Soap layer can
It is applied only to above water soluble film with selection.
The water soluble film is to meet that water is soluble to be facilitated in it with a thickness of 50 μm of conventional water soluble film added with flavouring agent
The bacteriostatic hand sanitizer of packaging is discharged.
The invention has the benefit that
(1) Traditional Chinese medicinal hand lotion primary raw material ingredient of the invention is Chinese medicine, not only harmless to human body and hand, Er Qiejing
It is good that overtesting detects its bactericidal effect；
(2) the detection test method of Traditional Chinese medicinal hand lotion of the invention can carry out the bacteriostasis rate of Traditional Chinese medicinal hand lotion accurate
It calculates, clearly defines the bactericidal effect of effective Traditional Chinese medicinal hand lotion and determine to provide for the detection of the bacteria-eliminating efficacy of Traditional Chinese medicinal hand lotion and cut
Real feasible detection test method.
(3) present invention is by the setting of pallet monomer so that the filling convenience and high-efficiency of bacteriostatic hand sanitizer, filling in hand cleanser
The intracorporal bacteriostatic hand sanitizer of chamber is 1-2ml, both can satisfy that disposable when washing one's hands to need in turn avoid conventional hand cleanser more
It is secondary to use bring bacterial cross-infection；
Detailed description of the invention
Fig. 1 hand cleanser production equipment of the present invention vacuumize before overall structure diagram；
Fig. 2 hand cleanser production equipment of the present invention vacuumize after overall structure diagram；
The overall structure diagram of Fig. 3 hand cleanser monomer of the present invention；
Specific embodiment
It is specifically described below with reference to Fig. 1-3 and 1 pair of table Traditional Chinese medicinal hand lotion and its production equipment of the invention.
Embodiment 1
1 Traditional Chinese medicinal hand lotion prepares material and instrument
1.1 material
Hand cleanser selection of the invention includes the honeysuckle for being purchased from hundred taste hall of Shandong, Fructus Forsythiae, picks up from the mulberry of Shandong Xiajin County
Leaf, purchased from the peppermint of the Shandong shop of Chinese medicines Jian Lian, chlorhexidine acetate (Mike woods, lot number C10122846), 20% gluconic acid chlorine oneself
Fixed (Mike woods, lot number C10105541), phosphate buffer PBS (Hyclone, lot number AD18274264), soybean casein fine jade
Rouge culture medium (Solarbio, lot number LA0170), sabouraud's agar (Solarbio, lot number S9710), Candida albicans,
Escherichia coli, staphylococcus aureus, batch cultur ware (ASONE), water are ultrapure water, and ethyl alcohol etc. is that analysis is pure.
1.2 instrument
Experiment of the invention is all made of following instrument, the BP211D type electronic balance of German Sai Duolisi group, and Ningbo is new
The SB-4200DTS type ultrasonic wave double frequency cleaning machine of sesame biotech inc, the new Jiangnan instrument in Ningbo Science and Technology Park area
The HWS-280 type constant temperature and humidity incubator of Co., Ltd, the type mold incubator of the Ningbo MJX-268 Jiangnan instrument plant, Su Jingan
Safe J16010024 type clean work station, the GZX-9023MBE type electric heating drum of Shanghai Medical Equipment Plant of Bo Xun Industrial Co., Ltd.
Wind drying box, the YB-169 type autoclave of Zhi Wei company of the U.S., the R-210 type Rotary Evaporators of BUCHI Labortechnik AG of Switzerland, Juan
The KDM type control-temperature electric heating set of city Hua Lu electric heating Instrument Ltd..
The preparation of 2 hand cleanser
Extracting honeysuckle, Fructus Forsythiae, mulberry leaf, peppermint crush, and cross No. 3 sieves；Accurately weighed honeysuckle 0.5g, Fructus Forsythiae 0.5g, mulberry leaf
50% ethyl alcohol 50mL reflux 1h is added in 0.1g, peppermint 0.5g, mixing, and extracting solution is cooled to room temperature, and 4000rpm is centrifuged 10min,
Dehydrated alcohol 100mL, ultrasonic 30min, supersonic frequency 40kHz, ultrasonic power 400W, mistake is added to doing in filtering, recycling design
Filter.14mL filtrate is taken, 14mL dehydrated alcohol is added, 12mL water mixes, 25mg chlorhexidine acetate is added according to every 10mL solution,
The ratio of 125 μ L glucose acid Chlorhexidines prepares hand cleanser sample to be tested, mixes to get bacteriostatic hand sanitizer of the invention.
3 bacteriostatic tests
3.1, by instrument apparatus high pressure sterilization, take three test tubes, mark Candida albicans, Escherichia coli, golden yellow Portugal respectively
Grape coccus.In the test tube of label Candida albicans, SDB culture medium 5mL and Candida albicans 5mL is added；In label large intestine bar
In the test tube of bacterium, TSB culture medium 5mL and Escherichia coli 0.5mg is added；In the test tube of label staphylococcus aureus, it is added
TSB 5mL and staphylococcus aureus 0.5mL.
Two test tubes are taken, mark TSB, SDB as reference substance respectively.TSB5mL, Yu Biao is added in the test tube of label TSB
Remember that SDB 5mL is added in the test tube of SDB is placed in five test tubes in constant incubator.After viable bacteria, with PBS, (phosphate is slow
Rush solution) 1000 times of dilution, it is spare.
The small filter paper of 3.2 1 × 1cm of preparation is several, is respectively put into label Candida albicans, Escherichia coli, golden yellow Portugal
In the culture dish of grape coccus, it is spare that corresponding diluted strain 20 μ L, 37 DEG C of drying 30min is added dropwise in each small filter paper.
3.3 take 32 culture dishes to mark Escherichia coli, staphylococcus aureus, Candida albicans, Escherichia coli+medicine respectively
Liquid, staphylococcus aureus+medical fluid, Candida albicans+medical fluid 2min, 5min, 10min, 20min, control group TSA (Sharpe fine jade
Rouge culture medium) and SDA (soybean casein agar) 2min, 5min, 10min, 20min respectively, it is added in culture medium prepared
TSA or SDA rotates culture dish, keeps its full and uniform, places solidification.
3.4 are put into the scraps of paper of drying in the cillin bottle containing medical fluid or PBS (phosphate buffered saline solution), start timing,
Respectively when acting on 2min, 5min, 10min, 20min, print is put into the cillin bottle containing 5mL PBS with sterile tweezer,
It mixes, draws 1mL respectively in the corresponding culture dish solidified, smoothened with spreading rod, culture dish is overturn after agar solidification,
It sets in constant incubator, makees the counting of viable bacteria bacterium colony.
4 experimental results
Candida albicans culture 48h takes out culture dish after Escherichia coli, staphylococcus aureus culture for 24 hours, by formula:
X=(A-B)/A × 100%
X --- bacteriostasis rate (%)
A --- control sample average colony number
B --- subject sample average clump count
The average colony number for calculating culture dish on same dilution, the results are shown in Table 1.
1 hand cleanser different role time bacteriostasis rate (%) of table
Embodiment 2
In conjunction with attached drawing 1, including pallet monomer 1, the recessed of tray surface is arranged in Traditional Chinese medicinal hand lotion production equipment of the invention
Slot 2 and convex block 3；It is characterized in that the pallet monomer 1 be can vacuum-packed enclosed construction, bottom is provided with several true
Suction stomata 4, vacuum suction hole 4 can be to vacuumizing inside pallet monomer 1；The upper surface of pallet monomer 1 is placed with film 5, thin
Film 5 is laid on pallet monomer 1 by the support of convex block 3；When the vacuum suction hole 4 vacuumizes, the film 5 is in pallet monomer 1
Upper formation is wavy, and wavy 5 upper surface of film has hand cleanser cavity 501.When wherein arrow direction representative vacuumizes in Fig. 2
Air-flow direction.
The Traditional Chinese medicinal hand lotion production equipment further includes feeder pipe 6, and feeder pipe 6 is needle-shaped injection device, and lifting rate is big
Small is every drop 1ml or so.
Since above-mentioned bacteriostatic hand sanitizer of the invention is Traditional Chinese medicinal hand lotion, only needed down when filling bacteriostatic hand sanitizer every time
6 blanking 1 to 2 of expects pipe drop；The volume of the hand cleanser cavity 501 is 2ml.
The hand cleanser cavity 501 is that substantially semi-cylindrical, the 1 upper surface convex block 3 of pallet monomer are triangular bump,
The top of each triangular bump 3 is on sustained height, after filling hand cleanser is in hand cleanser cavity 501, by water soluble film 7
It is covered on 5 top of film, seals to form several hand cleanser monomers 8 in its contact point using heat sealing machine.
In conjunction with attached drawing 2-3, soap layer 801, the thickness of soap layer 801 may further be coated on the hand cleanser monomer 8
Preferably 200 μm.Soap layer 801, which can choose, is applied only to 7 top of water soluble film.
The water soluble film 7 is to meet that water is soluble to facilitate it with a thickness of 50 μm of conventional water soluble film added with flavouring agent
The bacteriostatic hand sanitizer of inner packing is discharged.
The present invention is by the setting of pallet monomer 1 so that the filling convenience and high-efficiency of bacteriostatic hand sanitizer, filling in sap cavity of washing one's hands
Bacteriostatic hand sanitizer in body 501 is 1-2ml, both can satisfy that disposable when washing one's hands to need in turn avoid conventional hand cleanser more
It is secondary to use bring bacterial cross-infection；The setting coating soap layer 801 on hand cleanser monomer 8, when washing one's hands, hand grasping is washed
Hand liquid monomer 8 is placed in water source, and contact soap layer 801 can clean the dirt of hand, after hand dirt removal, soap first
Layer 801 disappears, and the exposure of water soluble film 7 on hand cleanser monomer 8, further water soluble film 7 is met water dissolution and will be wrapped in hand cleanser monomer 8
The bacteriostatic hand sanitizer of dress is discharged, and carries out deep layer sterilization and washes one's hands.
The above, only the preferred embodiment of the present invention, but scope of protection of the present invention is not limited thereto, it is any
Those familiar with the art in the technical scope disclosed by the present invention, can easily think of the change or the replacement, and should all contain
Lid is within protection scope of the present invention.Therefore, protection scope of the present invention should be based on the protection scope of the described claims.
Claims (7)
1. a kind of preparation method of Traditional Chinese medicinal hand lotion, which is characterized in that the preparation process of the hand cleanser is as follows: step (a) takes
Honeysuckle, Fructus Forsythiae, mulberry leaf, peppermint crush, and cross No. 3 sieves of 24 mesh；Accurately weighed 0.5 g of honeysuckle, Fructus Forsythiae 0.5g, mulberry leaf
0.1g, peppermint 0.5g, and it is mixed to form mixture；
Said mixture is added 50% ethyl alcohol 50mL reflux 1h and obtains extracting solution by step (b), and extracting solution is cooled to room temperature,
Revolving speed is centrifuged 10 min in the case where being 4000 rpm, filters extracting solution again;
100 mL of dehydrated alcohol is added to doing in step (c) recycling design, is ultrasonically treated 30 min, 40 kHz of supersonic frequency, ultrasound
400 W of power, is obtained by filtration filtrate again；
Step (d) takes the above-mentioned filtrate of 14 mL, 14 mL dehydrated alcohols, 12 mL water is added, and mix, adds according to every 10 mL solution
Enter the ratio of 25 mg chlorhexidine acetates, 125 μ L glucose acid Chlorhexidines, prepare hand cleanser, up to Traditional Chinese medicinal hand lotion after mixing
Stoste.
2. a kind of preparation method of Traditional Chinese medicinal hand lotion according to claim 1, it is characterised in that carried out in above-mentioned steps (a)
Before, first honeysuckle, Fructus Forsythiae, mulberry leaf, peppermint are carried out respectively to clean it is spare, to Fructus Forsythiae carry out lyophilization, to honeysuckle, mulberry
Leaf, peppermint carry out hot drying.
3. the preparation method of Traditional Chinese medicinal hand lotion according to claim 2, it is characterised in that lyophilization is being carried out to Fructus Forsythiae,
After carrying out hot drying to honeysuckle, mulberry leaf, peppermint, by obtained lyophilization Fructus Forsythiae and heat drying honeysuckle, mulberry leaf, peppermint
Ultramicro-powder is made by multistage plant ultra-micropowder grinding technology respectively.
4. the preparation method of Traditional Chinese medicinal hand lotion according to claim 2, it is characterised in that honeysuckle, mulberry leaf, peppermint into
The drying temperature of row heat drying is 120 DEG C or so.
5. the preparation method of Traditional Chinese medicinal hand lotion according to claim 2, it is characterised in that using freeze drier to Fructus Forsythiae
Carry out lyophilization.
6. the preparation method of Traditional Chinese medicinal hand lotion according to claim 3, it is characterised in that in above-mentioned preparation method, extracting solution
The process being cooled to room temperature carries out in two stages, and the first stage is that water-bath holding stage continues 1h or so；Second stage is fast
Fast cooling stage guarantees to be rapidly cooled to room temperature in 3min.
7. a kind of detection test method of Traditional Chinese medicinal hand lotion according to claim 1, it is characterised in that the Chinese medicine is washed one's hands
The detection test method of liquid the following steps are included:
Step 1: by instrument apparatus high pressure sterilization, taking three test tubes, mark Candida albicans, Escherichia coli, golden yellow Portugal respectively
Grape coccus；In the test tube of label Candida albicans, 5 mL of SDB culture medium and 5 mL of Candida albicans is added；In label large intestine
In the test tube of bacillus, 5 mL of TSB culture medium and 0.5 mg of Escherichia coli is added；In the test tube of label staphylococcus aureus,
TSB 5mL and staphylococcus aureus 0.5mL is added；
Step 2: taking two test tubes, mark TSB, SDB as reference substance respectively；TSB 5mL is added in the test tube of label TSB,
5 mL of SDB is added in the test tube of label SDB five test tubes are placed in constant incubator；After viable bacteria, with PBS (phosphoric acid
Salt buffer solution) 1000 times of dilution, it is spare；
Step 3: the small filter paper of 1 × 1 cm of preparation is several, is respectively put into label Candida albicans, Escherichia coli, golden yellow Portugal
In the culture dish of grape coccus, it is spare that corresponding diluted strain 20 μ L, 37 DEG C of 30 min of drying is added dropwise in each small filter paper；
Step 4: 32 culture dishes being taken to mark Escherichia coli, staphylococcus aureus, Candida albicans, Escherichia coli+medicine respectively
Liquid, staphylococcus aureus+medical fluid, 2 min of Candida albicans+medical fluid, 5 min, 10 min, 20 min, control group TSA are (husky
Family name's agar medium) and SDA (soybean casein agar) 2 min, 5 min, 10 min, 20 min respectively, it is added in culture medium
Prepared TSA or SDA rotates culture dish, keeps its full and uniform, places solidification；
Step 5: the scraps of paper of drying are put into the cillin bottle containing medical fluid or PBS (phosphate buffered saline solution), timing is started, point
Not when acting on 2 min, 5 min, 10 min, 20 min, print is put into the cillin bottle containing 5 mL PBS with sterile tweezer
In, it mixes, draws 1 mL respectively in the corresponding culture dish solidified, smoothened with spreading rod, training is overturn after agar solidification
Ware is supported, is set in constant incubator, the counting of viable bacteria bacterium colony is made；
Step 6: carrying out experimental result calculating.
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