The specific embodiment
Below by specific embodiment, nano silver antibacterial handwashing liquid of the present invention is further described, but not as limitation of the present invention.
Embodiment 1:
Nano silver antibacterial handwashing liquid of the present invention, it be take deionized water and is carrier, and nanometer silver, surfactant, wetting agent, toner and flavoring agent are dissolved in deionized water and make.Wherein, the percentage by weight of described nanometer silver, surfactant, wetting agent, toner, flavoring agent and deionized water is respectively:
Nanometer silver: 0.0003%;
Surfactant: alkylpolyoxyethylene sodium sulfate (AES) 2.0%, dodecyl sodium sulfate (LAS) 18.0% and coconut oil alkyl diethanolamide (6501) 8%;
Wetting agent: glycerol 1%;
Toner: pearling agent 1%, apple green 0.01%;
Flavoring agent: essence 0.6%;
Surplus is deionized water.
Embodiment 2:
Nano silver antibacterial handwashing liquid of the present invention, it be take deionized water and is carrier, and nanometer silver, surfactant, wetting agent, toner and flavoring agent are dissolved in deionized water and make.Wherein, the percentage by weight of described nanometer silver, surfactant, wetting agent, toner, flavoring agent and deionized water is respectively:
Nanometer silver: 0.0020%;
Surfactant: alkylpolyoxyethylene sodium sulfate (AES) 8.0%, dodecyl sodium sulfate (LAS) 12.0% and coconut oil alkyl diethanolamide (6501) 8%;
Wetting agent: glycerol 2%;
Toner: pearling agent 1%, apple green 0.01%;
Flavoring agent: essence 0.6%;
Surplus is deionized water.
Implementation column 3:
Nano silver antibacterial handwashing liquid of the present invention, it be take deionized water and is carrier, and nanometer silver, surfactant, wetting agent, toner and flavoring agent are dissolved in deionized water and make.Wherein, the percentage by weight of described nanometer silver, surfactant, wetting agent, toner, flavoring agent and deionized water is respectively:
Nanometer silver: 0.0030%;
Surfactant: alkylpolyoxyethylene sodium sulfate (AES) 10.0%, dodecyl sodium sulfate (LAS) 12.0% and coconut oil alkyl diethanolamide (6501) 8%;
Wetting agent: glycerol 2%;
Toner: pearling agent 1%, apple green 0.01%;
Flavoring agent: essence 0.6%;
Surplus is deionized water.
Implementation column 4:
Nano silver antibacterial handwashing liquid of the present invention, it be take deionized water and is carrier, and nanometer silver, surfactant, wetting agent, toner and flavoring agent are dissolved in deionized water and make.Wherein, the percentage by weight of described nanometer silver, surfactant, wetting agent, toner, flavoring agent and deionized water is respectively:
Nanometer silver: 0.0050%;
Surfactant: alkylpolyoxyethylene sodium sulfate (AES) 10.0%, dodecyl sodium sulfate (LAS) 15.0% and coconut oil alkyl diethanolamide (6501) 8%;
Wetting agent: glycerol 1%;
Toner: pearling agent 1%, apple green 0.01%;
Flavoring agent: essence 0.6%;
Surplus is deionized water.
Nano silver antibacterial handwashing liquid of the present invention can be prepared as follows, and it comprises following processing step:
(A) take alkylpolyoxyethylene sodium sulfate (AES), dodecyl sodium sulfate (LAS), add deionized water dissolving, obtains solution 1., standby;
(B) coconut oil alkyl diethanolamide (6501), pearling agent are joined in glycerol, obtain solution 2., standby;
(C) after 2. 1. solution stirred with solution, add nano-silver water solution, essence and apple green, stir, regulate pH value with NaOH, add deionized water to enough, finally make the nano silver antibacterial handwashing liquid.
For further verifying bacteriostasis and sterilization effect of the present invention, the inventor has carried out kill pathogenic fungus exam ine for above-mentioned nano silver antibacterial handwashing liquid.
Experiment material:
1. experimental strain:
Staphylococcus aureus ATCC 6538, escherichia coli 44148
Candida albicans ATCC10231
2. culture medium raw material:
Nutrient broth: lot number 000124, biochemical reagents, Beijing bispin microbiological culture media products factory
Nutrient agar: lot number 1001066, Beijing Luqiao Technology Co., Ltd.
Anhydrous glucose: lot number 990201, analytical pure, Tianjin Chemical Reagents Factory No.1
Peptone: lot number 20000215, biochemical reagents, Beijing bispin microbiological culture media products factory
Agar: Biotechnology Grad From Japan
Attached: the preparation of culture medium
(1) nutrient broth medium:
The 18g nutrient broth is dissolved in the 1000ml distilled water, heating for dissolving, and autoclaving, standby.
(2) nutrient agar:
The 33g Nutrient agar is dissolved in the 1000ml distilled water, heating for dissolving, and autoclaving, standby.
(3) Sharpe fluid medium:
Peptone 10g, anhydrous glucose 40g, be dissolved in the 1000ml distilled water, heating for dissolving, autoclaving, standby.
(4) husky fort agar culture medium:
Peptone 10g, anhydrous glucose 40g, be dissolved in the 300ml distilled water; 20g agar is dissolved in the 700ml distilled water; By the two mixing, autoclaving, standby.
3. sodium hydrogen phosphate: analytical pure, Tianjin Fengchuan Chemical Reagent Science & Technology Co., Ltd.
Potassium dihydrogen phosphate: analytical pure, the upright chemical reagent work of the Tianjin North Star
Sodium thiosulfate: analytical pure, Beijing chemical reagents corporation
Tween 80: Hunan Ning Kang pharmaceutical Co. Ltd
Lecithin: biochemical reagents, Tianjin recovery fine chemistry industry institute
4. inoculating loop, suction pipe, test tube, stopwatch, 37 ℃ of incubators etc.
Experimental technique and result
1. bacteriostasis rate test
Test organisms 24h slant culture is washed down with PBS, and alkene is interpreted into the bacteria suspension that concentration is 3 * 106 cfu/ml.Taking liquid 5ml 1 pipe; Get above-mentioned bacteria suspension 0.1ml and be placed in drug liquid tube, start timing, effect 2,5,10,20min, pipette respectively solution 0.5ml input in drug liquid tube with aseptic straw and contain 4.5ml nertralizer 2 in vitro, fully mixes, act on 10 minutes, suitably alkene is released, and draws respectively 1ml and is placed in two plates, pours into cool nutrient agar (for coliform, golden Portugal bacterium) or sabouraud's agar (for reading in vain bacterium) 20ml to 60 ℃, rotate plate, make it full and uniform, it is dull and stereotyped that agar solidifies rear upset, cultivates 48h for 37 ℃.Do the viable bacteria colony counting.With distilled water, replace medicinal liquid to do identical processing as the positive control pipe.
Calculate bacteriostasis rate:
X=(A mono-B)/A
X: bacteriostasis rate, %
A: the average clump count of positive control solution
B: tested medicinal liquid sample average clump count
Experimental result is as follows:
Table 1 nano silver antibacterial handwashing liquid (15 μ g/ml) bacteriostasis rate experimental result
|
2min |
5min |
10min |
20min |
Escherichia coli |
100% |
100% |
100% |
100% |
Staphylococcus aureus |
100% |
100% |
100% |
100% |
Candida albicans |
100% |
100% |
100% |
100% |
Nano silver antibacterial handwashing liquid 15 μ g/ml are to escherichia coli, staphylococcus aureus, and Candida albicans 2min, 5min, 10min, the 20min bacteriostasis rate is 100%.
2. sterilizing rate test
Test organisms 24h slant culture is washed down with PBS, and alkene is interpreted into the bacteria suspension that concentration is 3 * 108cfu/ml.Taking liquid 4ml 1 pipe adds above-mentioned bacteria suspension 0.5ml; Start timing, effect 2,5,10,20min, pipette respectively medicinal liquid 0.5ml input with aseptic straw and contain 4.5ml nertralizer 2 in vitro, fully mix, act on 10 minutes, suitably alkene is released, draw respectively 1ml and be placed in two plates, with cool nutrient agar (for coliform, golden Portugal bacterium) or sabouraud's agar (for reading in vain bacterium) 20ml to 60 ℃, pour into, rotate plate, make it full and uniform, it is dull and stereotyped that agar solidifies rear upset, cultivates 48h for 37 ℃.Do the viable bacteria colony counting.With distilled water, replace medicinal liquid to do same treatment, as positive control.Calculate sterilizing rate:
X=(A mono-B)/A
X: sterilizing rate, %;
A: the average clump count of control sample; B: tested sample average clump count
Table 2 nano silver antibacterial handwashing liquid (15 μ g/ml) sterilizing rate experimental result
|
2min |
5min |
10min |
20min |
Escherichia coli |
33.8% |
41.5% |
92.4% |
100% |
Staphylococcus aureus |
99.5% |
100% |
100% |
100% |
Candida albicans |
90.8% |
95.85% |
98.05% |
100% |
Visible by above result, nano silver antibacterial handwashing liquid 15 μ g/ml are to escherichia coli, staphylococcus aureus, and the sterilizing rate after Candida albicans 2min all is greater than 90%.
For verifying further whether the present invention can produce anaphylaxis, and the inventor has carried out hypersensitive test for above-mentioned nano silver antibacterial handwashing liquid.
Experiment material:
Healthy albino guinea-pig: skin is intact, and 48, male and female half and half, body weight 200-300g, be purchased from Mountain Western Medicine S University's Experimental Animal Center (animal number: SCXK (Shanxi) 2009-0001);
The nano silver antibacterial handwashing liquid: Zhejiang three standing grain nanosecond science and technology company limiteies provide;
DNFB: Tianjin recovery fine chemistry industry institute.
Method and step:
Cavia porcellus is divided into to 3 groups at random, 16 every group, is respectively test group, negative control group and positive controls.In front 24 h of test, by guinea pig back both sides unhairing (3 cm * 3 cm).To the test group Cavia porcellus, give tested material nano silver antibacterial handwashing liquid and induce and excite processing.Positive controls gives positive sensitizer DNFB and induces and excite processing.Negative control group is only given the nano silver antibacterial handwashing liquid and is excited processing.
3.1 sensitization contact (Induction exposure) is got nano silver antibacterial handwashing liquid 0.5ml, is coated in depilation district, test group guinea pig back left side.Every Cavia porcellus sub-cage rearing, after 6h with the warm water cleaning tested material.7th, 14 days, in kind repeat respectively 1 time.Positive controls is with 0.1% DNFB 0.5ml, and experimental technique is the same.
3.2 excite contact time to give 14 d after tested medicine sensitization, test group and negative control group all are coated in depilation district, guinea pig back right side by tested material nano silver antibacterial handwashing liquid 0.5ml, positive controls is by 0.1% 2,4-dinitrochlorobenzene 0.5ml is coated in depilation district, guinea pig back right side, after 6 h with the warm water cleaning tested material, after 24h and 48h, observe dermoreaction, mark by table 1 pair dermoreaction.
Observe and evaluation of result:
According to table 1 " skin allergy standards of grading " scoring, calculate the reaction scoring meansigma methods of respectively organizing Cavia porcellus.The difference of reacting with the negative control group guinea pig skin according to test group, the skin allergy effect of judgement tested material nano silver antibacterial handwashing liquid, in anaphylactoid animal ratio calculating sensitization rate occurring, according to the intensity of table 2 " sensitization strength grading standard " judgement tested material nano silver antibacterial handwashing liquid sensitivity response.
Sensitization rate computing formula is: sensitization rate/%=[the animal number of cases of skin allergy positive (no matter degree weight)/this group animal subject sum] * 100%
Table 1 sensitization of skin reaction standards of grading
Erythema forms |
Scoring |
Edema forms |
Scoring |
Without erythema |
0 |
Without edema |
0 |
Slight erythema |
1 |
Mild edema |
1 |
Moderate erythema |
2 |
Intermediate edema |
2 |
Serious erythema |
3 |
Serious edema |
3 |
Edematous erythema |
4 |
? |
? |
Table 2 sensitization strength grading standard
Sensitization rate (%) |
Sensitization intensity |
0~8 |
Extremely slight |
9~28 |
Slightly |
29~64 |
Moderate |
65~80 |
Intensity |
81~100 |
Pole strength |
The sensitization of skin experimental result of table 3 nano silver antibacterial handwashing liquid
Brief summary and discussion
The sensitization of skin experimental result of nano silver antibacterial handwashing liquid can find out, the nano silver antibacterial handwashing liquid has extremely slight sensitization.
For further whether checking the present invention is irritant to skin, the inventor has carried out irritation test for above-mentioned nano silver antibacterial handwashing liquid.
Experiment material:
12 of healthy rabbits, body weight 2 scholar 0.2kg, male and female half and half, be purchased from Mountain Western Medicine S University's zoopery center; Nano silver antibacterial handwashing liquid: provided by Zhejiang three standing grain nanosecond science and technology company limiteies.
Experimental technique:
1. single-dose irritation testGet 8 of healthy rabbits, be divided at random 2 groups, 4 every group.Getting wherein 1 group is the intact skin test group, tests front 24 h, by rabbit spinal column both sides unhairing (3 cm * 3 cm), respectively as tested material district and negative control area.Before giving tested material nano silver antibacterial handwashing liquid, should check whether skin of unhairing has mechanical damage, diminishing the wounded should not test.Tested material nano silver antibacterial handwashing liquid 0.5 ml is smeared in tested material next day district, covers with double gauze (2. 5 cm * 2. 5 cm) and polyethylene film, is fixed with nonirritant adhesive plaster and binder; Negative control area is as blank.Get other 1 group of rabbit, before being coated with tested material nano silver antibacterial handwashing liquid, on the skin of unhairing of 2.5cm * 2.5cm,, sterilization clean with 75% ethanol exposes skin, after the ethanol volatilization, the damaged wound of standardized " well " shape in dermatotome with sterilizing blade or injection needle, as the damaged skin test group, the drug treating mode is with the intact skin group.After 4 h, water washes away all rabbit depilations district residue, respectively at 1h, 24h and 48h after the removal tested material, observe the local skin reaction, by table 1, carry out the irritant reaction scoring, and press the rank of table 2 evaluation nano silver antibacterial handwashing liquid to the animal skin stimulus intensity.
. multiple dosing intact skin irritation test4 of healthy rabbits, grouping, unhairing, medication are identical with the single-dose irritation test.4h after smearing, water cleans, and removes residue.Smear every day once, smear continuously 14d.24h observed result after at every turn smearing, press table 1 scoring, and press the rank that table 2 is judged skin irritation intensity.
Table 1 skin irritation reaction standards of grading
Erythema forms |
The reaction scoring |
Edema forms |
The reaction scoring |
Nothing |
0 |
Nothing |
0 |
Visible reluctantly |
1 |
Visible reluctantly |
1 |
Obviously |
2 |
Cutaneous protuberance, profile is clear |
2 |
Seriously |
3 |
The about 1mm of edema protuberance |
3 |
The aubergine erythema, and eschar is arranged |
4 |
The edema protuberance surpasses 1mm |
4 |
Table 2 skin irritation intensity evaluation standard
Skin irritation index |
The stimulus intensity rank |
0~<0.5 |
Nonirritant |
0.5~<2.0 |
Subexcite |
2.0~<6.0 |
Medium zest |
6.0~8.0 |
Strong and stimulating |
Experimental result
Visible by table 3 result: after nano silver antibacterial handwashing liquid rabbit single-dose, intact skin and damaged skin are showed no local erythema and edema forms, and relatively without significant difference, is showed no zest with the blank group.Multiple dosing the results are shown in Table 4, and nano silver antibacterial handwashing liquid successive administration 14d has no local erythema and edema and forms, and with the blank group, compares without significant difference, all nonirritant.
Table 3 nano silver antibacterial handwashing liquid single-dose skin irritation test result
Table 4 nano silver antibacterial handwashing liquid multiple dosing intact skin stimulation test result
Brief summary and discussion
Visible by above result, the nano silver antibacterial handwashing liquid is without skin irritation.
The above specific embodiment is only the preferred embodiment of this creation, not in order to limit this creation, any modification of making within all spirit in this creation and principle, is equal to replacement, improvement etc., within all should being included in the protection domain of this creation.