The specific embodiment
Below by specific embodiment nano silver antibacterial handwashing liquid of the present invention is further specified, but not as limitation of the present invention.
Embodiment 1:
Nano silver antibacterial handwashing liquid of the present invention, it is take deionized water as carrier, nanometer silver, surfactant, wetting agent, toner and flavoring agent is dissolved in the deionized water makes.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 is take deionized water as carrier, nanometer silver, surfactant, wetting agent, toner and flavoring agent is dissolved in the deionized water makes.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 is take deionized water as carrier, nanometer silver, surfactant, wetting agent, toner and flavoring agent is dissolved in the deionized water makes.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 is take deionized water as carrier, nanometer silver, surfactant, wetting agent, toner and flavoring agent is dissolved in the deionized water makes.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 by weighing alkylpolyoxyethylene sodium sulfate (AES), dodecyl sodium sulfate (LAS) adds deionized water dissolving, obtains solution 1., and is for subsequent use;
(B) coconut oil alkyl diethanolamide (6501), pearling agent are joined in the glycerol, obtain solution 2., for subsequent use;
(C) 2. 1. solution stirred with solution after, add nano-silver water solution, essence and apple green, stir, regulate pH value with NaOH, add deionized water to capacity, 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, for subsequent use.
(2) nutrient agar:
The 33g Nutrient agar is dissolved in the 1000ml distilled water, heating for dissolving, and autoclaving, for subsequent use.
(3) Sha Shi fluid medium:
Peptone 10g, anhydrous glucose 40g is dissolved in the 1000ml distilled water, heating for dissolving, autoclaving, for subsequent use.
(4) husky fort agar culture medium:
Peptone 10g, anhydrous glucose 40g is dissolved in the 300ml distilled water; 20g agar is dissolved in the 700ml distilled water; With the two mixing, autoclaving, for subsequent use.
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 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 place drug liquid tube, the beginning timing, effect 2,5,10,20min pipettes respectively in the drug liquid tube solution 0.5ml with aseptic straw and drops into and contain 4.5ml nertralizer 2 in vitro, fully mixing, act on 10 minutes, suitably alkene is released, and draws respectively 1ml and places two plates, pours into cool nutrient agar (being used for coliform, golden Portugal bacterium) or sabouraud's agar (being used 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.Replace medicinal liquid to do identical processing as the positive control pipe with distilled water.
Calculate bacteriostasis rate:
X=(A one 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 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; The beginning timing, effect 2,5,10,20min pipettes respectively medicinal liquid 0.5ml input with aseptic straw and contains 4.5ml nertralizer 2 in vitro, fully mixing acts on 10 minutes, and suitably alkene is released, draw respectively 1ml and place two plates, nutrient agar (being used for coliform, golden Portugal bacterium) or sabouraud's agar (being used for reading in vain bacterium) 20ml with cold to 60 ℃ pour into, and the rotation plate makes it full and uniform, it is dull and stereotyped that agar solidifies rear upset, cultivates 48h for 37 ℃.Do the viable bacteria colony counting.Replace medicinal liquid to do same treatment with distilled water, as positive control.Calculate sterilizing rate:
X=(A one 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% |
By above result as seen, nano silver antibacterial handwashing liquid 15 μ g/ml are to escherichia coli, staphylococcus aureus, and the sterilizing rate behind the Candida albicans 2min is all 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 is 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 3 groups at random, 16 every group, is respectively test group, negative control group and positive controls.In front 24 h of test, with 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, behind the 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 the tested medicine sensitization, test group and negative control group all are coated in depilation district, guinea pig back right side with tested material nano silver antibacterial handwashing liquid 0.5ml, positive controls is with 0.1% 2,4-dinitrochlorobenzene 0.5ml is coated in depilation district, guinea pig back right side, behind 6 h with the warm water cleaning tested material, observe dermoreaction behind 24h and the 48h, 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.Difference according to test group and the reaction of negative control group guinea pig skin, judge the skin allergy effect of tested material nano silver antibacterial handwashing liquid, calculate the sensitization rate in anaphylactoid animal ratio occurring, judge the intensity of tested material nano silver antibacterial handwashing liquid sensitivity response according to table 2 " sensitization strength grading standard ".
Sensitization rate computing formula is: sensitization rate/%=[the animal example number 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 that the nano silver antibacterial handwashing liquid has extremely slight sensitization.
For verifying further whether the present invention has zest to skin, and 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 are purchased from Mountain Western Medicine S University 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, with rabbit spinal column both sides unhairing (3 cm * 3 cm), respectively as tested material district and negative control area.Should check whether skin of unhairing has mechanical damage before giving tested material nano silver antibacterial handwashing liquid, diminishing the wounded should not test.Next day, tested material nano silver antibacterial handwashing liquid 0.5 ml was smeared in the tested material district, covered with double gauze (2. 5 cm * 2. 5 cm) and polyethylene film, was 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, expose skin with 75% ethanol cleaning, sterilization, after the ethanol volatilization, the damaged wound of standardized " well " shape in the dermatotome with sterilization blade or injection needle, as the damaged skin test group, the drug treating mode is with the intact skin group.All rabbit depilation district residues of water flush away behind 4 h, observe the local skin reaction respectively at 1h, 24h and 48h after the removal tested material, carry out the irritant reaction scoring by table 1, and press table 2 evaluation nano silver antibacterial handwashing liquid to the rank of 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 is pressed table 1 scoring, and presses 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 |
Reluctantly as seen |
1 |
Reluctantly as seen |
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
By table 3 result as seen: behind the nano silver antibacterial handwashing liquid rabbit single-dose, intact skin and damaged skin are showed no local erythema and edema forms, and compares without significant difference with the blank group, is showed no zest.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 compares without significant difference with the blank group, 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
By above result as seen, the nano silver antibacterial handwashing liquid is without skin irritation.
The above specific embodiment only is the preferred embodiment of this creation, not in order to limiting this creation, all in this creation spirit and principle within make any modification, be equal to replacement, improvement etc., all should be included within the protection domain of this creation.