CN112602707B - Sebacic acid as ultraviolet sterilization synergist and application thereof - Google Patents
Sebacic acid as ultraviolet sterilization synergist and application thereof Download PDFInfo
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- CXMXRPHRNRROMY-UHFFFAOYSA-N sebacic acid Chemical compound OC(=O)CCCCCCCCC(O)=O CXMXRPHRNRROMY-UHFFFAOYSA-N 0.000 title claims abstract description 128
- 230000001954 sterilising effect Effects 0.000 title claims abstract description 48
- 238000004659 sterilization and disinfection Methods 0.000 title claims abstract description 40
- 230000001580 bacterial effect Effects 0.000 claims description 22
- 239000007788 liquid Substances 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 13
- 241000588724 Escherichia coli Species 0.000 claims description 7
- 230000001678 irradiating effect Effects 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 2
- 241000894006 Bacteria Species 0.000 abstract description 17
- 230000000694 effects Effects 0.000 abstract description 16
- 230000002195 synergetic effect Effects 0.000 abstract description 7
- 238000011160 research Methods 0.000 abstract description 6
- 230000009471 action Effects 0.000 abstract description 4
- 239000002609 medium Substances 0.000 description 25
- 229920001817 Agar Polymers 0.000 description 13
- 239000008272 agar Substances 0.000 description 13
- 238000012360 testing method Methods 0.000 description 12
- 239000006154 MacConkey agar Substances 0.000 description 9
- 241001360526 Escherichia coli ATCC 25922 Species 0.000 description 8
- 230000000844 anti-bacterial effect Effects 0.000 description 8
- 238000010790 dilution Methods 0.000 description 8
- 239000012895 dilution Substances 0.000 description 8
- 238000002474 experimental method Methods 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 239000002504 physiological saline solution Substances 0.000 description 6
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 5
- 238000011156 evaluation Methods 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 239000000843 powder Substances 0.000 description 5
- 206010059866 Drug resistance Diseases 0.000 description 4
- 230000002070 germicidal effect Effects 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- 241000208125 Nicotiana Species 0.000 description 3
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 3
- 230000003385 bacteriostatic effect Effects 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 150000007524 organic acids Chemical class 0.000 description 3
- 230000005855 radiation Effects 0.000 description 3
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- 238000007619 statistical method Methods 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 238000009631 Broth culture Methods 0.000 description 2
- 102000053602 DNA Human genes 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- WNLRTRBMVRJNCN-UHFFFAOYSA-N adipic acid Chemical compound OC(=O)CCCCC(O)=O WNLRTRBMVRJNCN-UHFFFAOYSA-N 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
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- 229920002477 rna polymer Polymers 0.000 description 2
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 description 2
- 238000009281 ultraviolet germicidal irradiation Methods 0.000 description 2
- 241000195493 Cryptophyta Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000010306 acid treatment Methods 0.000 description 1
- 235000011037 adipic acid Nutrition 0.000 description 1
- 239000001361 adipic acid Substances 0.000 description 1
- 125000001931 aliphatic group Chemical group 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 239000004359 castor oil Substances 0.000 description 1
- 235000019438 castor oil Nutrition 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
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- 235000019441 ethanol Nutrition 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- ZEMPKEQAKRGZGQ-XOQCFJPHSA-N glycerol triricinoleate Natural products CCCCCC[C@@H](O)CC=CCCCCCCCC(=O)OC[C@@H](COC(=O)CCCCCCCC=CC[C@@H](O)CCCCCC)OC(=O)CCCCCCCC=CC[C@H](O)CCCCCC ZEMPKEQAKRGZGQ-XOQCFJPHSA-N 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000000622 irritating effect Effects 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
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- 230000001937 non-anti-biotic effect Effects 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 244000045947 parasite Species 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
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- 238000001228 spectrum Methods 0.000 description 1
- TYFQFVWCELRYAO-UHFFFAOYSA-N suberic acid Chemical compound OC(=O)CCCCCCC(O)=O TYFQFVWCELRYAO-UHFFFAOYSA-N 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/02—Saturated carboxylic acids or thio analogues thereof; Derivatives thereof
- A01N37/04—Saturated carboxylic acids or thio analogues thereof; Derivatives thereof polybasic
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
- A61L2/02—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using physical phenomena
- A61L2/08—Radiation
- A61L2/10—Ultraviolet radiation
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Plant Pathology (AREA)
- Engineering & Computer Science (AREA)
- Dentistry (AREA)
- Pest Control & Pesticides (AREA)
- Agronomy & Crop Science (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
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Abstract
The invention belongs to the technical field of ultraviolet sterilization, and discloses an ultraviolet sterilization synergist sebacic acid and application thereof. The research is based on that the A wave band Ultraviolet (UVA) and the sebacic acid have weak sterilization effects (the sterilization amount is 0.5log-1.5log when the Ultraviolet (UVA) and the sebacic acid act independently), and the Ultraviolet (UVA) and the sebacic acid have obvious synergistic sterilization effects after combined action. Compared with the traditional ultraviolet sterilization technology, the ultraviolet sterilization device can kill bacteria more efficiently and reduce the influence of the quality of the ultraviolet lamp on the sterilization effect.
Description
Technical Field
The invention relates to the technical field of ultraviolet sterilization, and particularly relates to an ultraviolet sterilization synergist sebacic acid and application thereof.
Background
The uv sterilization technique is a convenient method with no chemical residue and little environmental impact, and is commonly used to disinfect gas, liquid and solid surfaces. Ultraviolet radiation is a generic term for radiation in the electromagnetic spectrum having a wavelength of 400nm to 10nm and does not cause human vision. It is invisible light having a higher frequency than bluish violet light, and ultraviolet rays are classified into UVA, UVB, UVC, and UVD. The ultraviolet lamp sterilization is used in various places due to the advantages of low cost, convenient use, no drug resistance and the like. In recent years, bacterial drug resistance is getting worse due to the use of a large amount of antibiotics, and the effect of treating infection caused by germs is gradually weakened along with the appearance of multi-drug resistant bacteria, so that the research on non-antibiotic sterilization technology is more and more applied in clinic, and the research on ultraviolet sterilization effect is more and more intensive.
The principle of the ultraviolet sterilization technology is that ultraviolet radiation with proper wavelength is utilized to destroy the molecular structure of DNA (deoxyribonucleic acid) or RNA (ribonucleic acid) in organism cells to cause death of growing cells and/or regenerating cells, thereby achieving the sterilization effect. The ultraviolet sterilization technology is based on modern epidemic prevention science, medicine and photodynamic, and specially designed ultraviolet with high efficiency, high strength and long service life is adopted to irradiate the surface of an object to directly kill various pathogens such as bacteria, viruses, parasites, algae and the like on the surface of the object. Meanwhile, bacteria can be killed under the condition of not generating antibiotic drug resistance, so that fine drug resistance is not improved. However, low-energy ultraviolet light by itself is not sufficient to achieve a very good germicidal effect, the germicidal time is long and the bacteria cannot be completely killed.
Sebacic acid is also known as n-sebacic acid, 10-sebacic acid, 1, 8-octanedioic acid. Sebacic acid belongs to aliphatic dibasic acid, has molecular mass of 202.25, and exists in flue-cured tobacco leaf, burley tobacco leaf and aromatic tobacco leaf. Sebacic acid is white flaky crystal at room temperature, and the industrial product is slightly yellow. Slightly soluble in water, insoluble in benzene, petroleum ether and carbon tetrachloride, and easily soluble in ethanol and diethyl ether. The sebacic acid is combustible and has low toxicity. It is harmful to oral administration and has irritant effects on eyes, respiratory system and skin. The natural castor oil or adipic acid monoester is used as raw material to prepare esters of sebacic acid, and the esters have wide application. In order to enhance the ultraviolet sterilization effect, the research of the ultraviolet sterilization effect synergist is an essential step. However, the currently reported organic acids still have little synergistic effect on ultraviolet sterilization, and a new ultraviolet sterilization synergist needs to be supplemented to increase the sterilization effect of ultraviolet rays in various occasions.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provide the application of sebacic acid as an ultraviolet sterilization synergist.
The second purpose of the invention is to provide a method for sterilizing by using sebacic acid in combination with ultraviolet.
The purpose of the invention is realized by the following technical scheme:
application of sebacic acid as ultraviolet sterilization synergist.
The research of the invention finds that the combination of the sebacic acid and the ultraviolet can obviously enhance the ultraviolet sterilization capability, so that the sebacic acid can be used as a synergist for the ultraviolet sterilization.
The invention also provides an ultraviolet sterilization synergist which comprises sebacic acid.
The invention also provides a method for sterilizing by combining ultraviolet and sebacic acid, wherein the irradiation time of the ultraviolet is 30min, and the concentration of the sebacic acid is 1 mM/L.
Preferably, in the method for sterilizing, the ultraviolet intensity is 2.4-3.0mW/cm2。
Preferably, in the sterilization method, the ultraviolet irradiation is performed for 30min before preheating.
Preferably, the method of sterilizing comprises the steps of:
(1) mixing the bacterial liquid with 1mM/L sebacic acid, placing the mixture in a six-hole plate, and setting ultraviolet irradiation conditions: wavelength range of 300-460nm, power of 18W, distance from the six-hole plate to the ultraviolet lamp tube of 8cm, and ultraviolet intensity of 2.4-3.0mW/cm2；
(2) Preheating the ultraviolet box for 20-40min, and placing the six-hole plate in the ultraviolet box for irradiating for 25-35 min.
More preferably, the final concentration of the bacterial liquid in the step (1) of the sterilization method is 106CFU/mL。
Compared with the prior art, the invention has the following beneficial effects:
the invention provides a synergist having a synergistic effect with ultraviolet, and the research is based on that the sterilization effect of Ultraviolet (UVA) in the A wave band and organic sebacic acid is weak (the sterilization amount is 0.5log-1.5log in the single action), and the synergistic sterilization effect is obvious after the synergistic sterilization effect is carried out on the ultraviolet and organic sebacic acid. Compared with the traditional ultraviolet sterilization technology, the ultraviolet sterilization device can kill bacteria more efficiently and reduce the influence of the quality of the ultraviolet lamp on the sterilization effect.
Detailed Description
The following further describes the embodiments of the present invention. It should be noted that the description of the embodiments is provided to help understanding of the present invention, but the present invention is not limited thereto. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
The test methods used in the following experimental examples are all conventional methods unless otherwise specified; the materials, reagents and the like used are, unless otherwise specified, commercially available reagents and materials.
The ultraviolet lamp used was brand PHILIPS TL-D; wattage: 18W; voltage: 220V; wavelength: (UVA)300-469 nm; tube diameter of the lamp tube: 25 mm; length: 60 cm. The sebacic acid is a biological source leaf brand with the purity of 99%.
The judgment principle is as follows: for example, the bacterial quantity reduced by the experimental group is reduced by 2 log values on the basis of the bacterial quantity reduction of the ultraviolet treatment alone and the organic acid treatment alone, namely the organic acid and the ultraviolet have the synergistic bactericidal effect.
Example 1 germicidal Effect of different UV irradiation times on ATCC25922 Strain
1. Experimental materials:
(1) the ultraviolet lamp used for the test is PHILIPS TL-D brand; wattage: 18W; voltage: 220V; wavelength: (UVA)300-469 nm; tube diameter of the lamp tube: 25 mm; length: 60 cm.
(2) Test medium: the MH agar medium and the MacConkey agar medium (purchased from Guangdong Huancao Microscience Co., Ltd.) which were sterilized by autoclaving were cooled to 40 ℃, 20mL of the MH agar medium and the MacConkey agar medium were put into a sterile petri dish using a pipette, and naturally dried for 30min to obtain the MH agar medium and the MacConkey agar medium.
Coli standard strain ATCC25922 (laboratory collection).
2. Preparation work before the test:
(1) starting an ultraviolet lamp and continuously irradiating for 30 minutes for preheating;
(2) the E.coli standard strain ATCC25922 was cultured on MacconyKa medium to a suitable size.
3. Ultraviolet sterilization effect evaluation experiment:
(1) inoculating Escherichia coli ATCC25922, placing single colony in a centrifuge tube filled with 4mL MH broth, putting the centrifuge tube in a 37-degree shaking table, incubating for 4 hours at 180 revolutions, and taking out the centrifuge tube;
(2) placing the centrifuge tube in a centrifuge, centrifuging for 8min at 5000 rpm, pouring out supernatant, adding equal volume of physiological saline for resuspension, and performing gradient dilution to obtain final bacterial load of 106CFU/mL；
(3) Adding 1mL of bacterial liquid into a six-hole plate;
(4) setting blank control group and ultraviolet irradiation treatment group, wherein the ultraviolet irradiation treatment group is divided into three groups, and irradiating for 15min, 30min and 60min respectively.
(5) After the ultraviolet irradiation is finished, 100 mu L of bacterial liquid is absorbed and added into a 2mL centrifuge tube filled with 900 mu L of 0.85% physiological saline for gradient dilution, 25 mu L of bacterial liquid is absorbed and dropped on an MH agar culture medium after dilution, the culture box with the temperature of 37 ℃ is incubated for 16-18h, counting is carried out, and the statistical analysis is carried out after the experimental result is repeated through three biology.
The results are shown in table 1, the bacteriostatic effect of ultraviolet on bacteria is increased with the increase of the irradiation time, wherein the difference between the number of bacteria in the group irradiated with ultraviolet for 15min and the number of bacteria in the blank control group is not large, which indicates that the short irradiation time of ultraviolet cannot produce a significant bacteriostatic effect on ATCC 25922. The number of bacteria in the group irradiated by ultraviolet for 30min begins to decrease, which indicates that the ultraviolet begins to have a certain bacteriostatic effect on the bacteria.
TABLE 1 germicidal Effect of UV irradiation time on ATCC25922 strains
Example 2 determination of MIC of sebacic acid against ATCC25922 Strain
1. Experimental materials:
(1) and (3) testing: autoclaved MH broth was cooled for use.
Coli standard strain ATCC25922 (laboratory collection).
2. Preparation work before the test:
(1) 0.2043g of sebacic acid powder is weighed, a solvent is 10mL of absolute ethyl alcohol, the concentration of sebacic acid is 100mM/L, and the sebacic acid powder is dissolved and filtered by a filter membrane for later use;
(2) the E.coli standard strain ATCC25922 was cultured on MacconyKa agar medium to a suitable size.
3. Sebacic acid bactericidal effect evaluation experiment:
(1) inoculating Escherichia coli ATCC25922, placing the single colony in a centrifugal tube filled with 4mLMH broth, putting the centrifugal tube in a 37-degree shaking table, incubating for 4 hours at 180 revolutions, and taking out the centrifugal tube;
(2) coli after incubation was released 100-fold using MH broth, approximately 106CFU/mL for standby;
(3) taking a sterile 96-well plate, adding 180 mu of LMH broth culture medium into the 1 st well, and adding 100 mu of LMH broth culture medium into the 2 nd to 11 th wells;
(4) add 20. mu.L 100mM/L sebacic acid into column 1, blow and beat uniformly, suck 100. mu.L to well 2, and so on, suck 100. mu.L from well 10 and discard.
(5) Adding 100 mu L of diluted bacterium liquid into the 1 st to 11 th holes, and adding 200 mu L of MH broth into the 12 th hole;
(6) repeating the steps (3) to (5) for three times;
(7) and putting the inoculated 96-well plate into a 37-degree incubator for incubation for 16-18h, and reading the result.
As shown in Table 2, the Mic value of sebacic acid is more than 1mM/L, bacteria are not obviously inhibited from growing at the concentration of sebacic acid lower than the Mic value, and the concentration of sebacic acid of 1mM/L is finally selected as the use concentration of the ultraviolet synergist in consideration of high use cost of sebacic acid with high concentration and use of sebacic acid with low concentration which is usually diluted in practical application.
TABLE 2 MIC of sebacic acid against ATCC25922 strain and concentrations selected for the experiment
| Experimental strains | Sebacic acid Mic | Concentration for experiment |
| ATCC 25922 | >1mM/L | 1mM/L |
Example 3 bactericidal effect of sebacic acid on ATCC25922 strain after 30 minutes:
1. experimental materials:
(1) test medium: the MH agar medium and the MacConkey agar medium (purchased from Guangdong Huancao Microscience Co., Ltd.) which were sterilized by autoclaving were cooled to 40 ℃, 20mL of the MH agar medium and the MacConkey agar medium were put into a sterile petri dish using a pipette, and naturally dried for 30min to obtain the MH agar medium and the MacConkey agar medium.
Coli standard strain ATCC25922 (laboratory collection).
2. Preparation work before the test:
(1) (1) weighing 0.2043g of sebacic acid powder, dissolving 10mL of absolute ethyl alcohol as a solvent, and filtering the solution with a filter membrane for later use, wherein the concentration of the sebacic acid is 100 mM/L;
(2) the E.coli standard strain ATCC25922 was cultured on MacconyKa medium to a suitable size.
3. Sebacic acid bactericidal effect evaluation experiment:
(1) inoculating Escherichia coli ATCC25922, placing the single colony in a centrifugal tube filled with 4mLMH broth, putting the centrifugal tube in a 37-degree shaking table, incubating for 4 hours at 180 revolutions, and taking out the centrifugal tube;
(2) placing the centrifuge tube in a centrifuge, centrifuging for 8min at 5000 rpm, pouring out supernatant, adding equal volume of physiological saline for resuspension, and performing gradient dilution to obtain final bacterial load of 106CFU/mL；
(3) Adding 1mL of bacterial liquid into a six-hole plate, adding 10 mu L of 100mM/L sebacic acid, and uniformly mixing to obtain a final concentration of 1 mM/L;
(4) setting a control, a blank control and 1mM/L sebacic acid for 30 min;
(5) and (3) sucking 100 mu L of bacterial liquid, adding the bacterial liquid into a 2ml centrifuge tube filled with 900 mu L of 0.85% physiological saline, carrying out gradient dilution, sucking 25 mu L of the diluted bacterial liquid, dripping the diluted bacterial liquid on an MH agar culture medium, incubating the bacterial liquid in a 37-degree incubator for 16-18h, counting, and carrying out statistical analysis after three biological repetitions of experimental results.
As shown in Table 3, the number of bacteria in the blank control group was not significantly different from that in the group in which 1mM/L sebacic acid acted for 30 min; it was demonstrated that 1mM/L sebacic acid did not change the number of bacteria after 30min treatment with ATCC25922 bacteria. The bactericidal effect of 1mM/L sebacic acid on ATCC25922 is weak, so that sebacic acid with the concentration of 1mM/L and ultraviolet are used for 30min as final conditions.
TABLE 3 Bactericidal Effect of sebacic acid on ATCC25922 Strain
Example 4 evaluation of the killing effect of UV and UV potentiators on E.coli ATCC25922
1. Experimental materials:
(1) the ultraviolet lamp used for the test is PHILIPS TL-D brand; wattage: 18W; voltage: 220V; wavelength: (UVA)300-469 nm; tube diameter of the lamp tube: 25 mm; length: 60 cm.
(2) Test medium: the MH agar medium and the MacConkey agar medium (purchased from Guangdong Huancao Microscience Co., Ltd.) which were sterilized by autoclaving were cooled to 40 ℃, 20mL of the MH agar medium and the MacConkey agar medium were put into a sterile petri dish using a pipette, and naturally dried for 30min to obtain the MH agar medium and the MacConkey agar medium.
Coli standard strain ATCC25922 (laboratory collection).
2. Preparation work before the test:
(1) starting an ultraviolet lamp and continuously irradiating for 30 minutes for preheating;
(2) 0.2043g of sebacic acid powder is weighed, a solvent is 10mL of absolute ethyl alcohol, the concentration of sebacic acid is 100mM/L, and the sebacic acid powder is dissolved and filtered by a filter membrane for later use;
(3) the E.coli standard strain ATCC25922 was cultured on MacconyKa medium to a suitable size.
3. Ultraviolet synergist effect evaluation experiment:
(1) inoculating Escherichia coli ATCC25922, placing the single colony in a centrifugal tube filled with 4mLMH broth, putting the centrifugal tube in a 37-degree shaking table, incubating for 4 hours at 180 revolutions, and taking out the centrifugal tube;
(2) placing the centrifuge tube in a centrifuge, centrifuging for 8min at 5000 rpm, pouring out supernatant, adding equal volume of physiological saline for resuspension, and performing gradient dilution to obtain final bacterial load of 106；
(3) Adding 1mL of bacterial liquid into a six-hole plate, adding 10 mu L of 100mM/L sebacic acid, and uniformly mixing to obtain a final concentration of 1mM/L
(4) Setting a control, processing blank control and 1mM/L sebacic acid for shading, putting into an ultraviolet irradiation box, and irradiating ultraviolet for 30min by a combined action group and an ultraviolet irradiation processing group;
(5) after the ultraviolet irradiation is finished, 100 mu L of bacterial liquid is absorbed and added into a 2ml centrifuge tube filled with 900 mu L of 0.85% physiological saline for gradient dilution, 25 mu L of bacterial liquid is absorbed and dropped on an MH agar culture medium after dilution, the culture box with the temperature of 37 ℃ is incubated for 16-18h, counting is carried out, and the experimental result is subjected to statistical analysis after three biological repetitions.
In this experiment, a growth control, an ultraviolet control, a 1mM/L sebacic acid control, and a violet plus 1mM/L sebacic acid test were set, and the test was performed according to the method of example 1.
The results are shown in Table 4, where bacteria grew normally in the blank control group, indicating that E.coli ATCC25922 grew normally under the experimental conditions; under the irradiation of an ultraviolet lamp for 30min, the Escherichia coli ATCC25922 is not obviously reduced compared with a blank control group, which shows that the inhibition effect of the ultraviolet lamp on the Escherichia coli ATCC25922 is not obvious; the same results also show that 1mM/L sebacic acid has no significant inhibitory effect on Escherichia coli ATCC 25922; the combined action result of 30min shows that the synergistic effect of the ultraviolet rays and the 1mM/L sebacic acid has obvious bactericidal effect on Escherichia coli ATCC 25922.
TABLE 4 Bactericidal Effect of UV and sebacic acid combination on ATCC25922 Strain
The embodiments of the present invention have been described in detail, but the present invention is not limited to the described embodiments. It will be apparent to those skilled in the art that various changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, and the scope of protection is still within the scope of the invention.
Claims (4)
1. The method for sterilizing by combining ultraviolet with sebacic acid is characterized in that the irradiation time of ultraviolet is 30min, the concentration of sebacic acid is 1mM, and the ultraviolet intensity is 2.4-3.0 mW/ cm2The sterilization object is an Escherichia coli standard strain ATCC 25922.
2. The method for sterilizing by combining ultraviolet and sebacic acid according to claim 1, wherein the ultraviolet chamber is preheated for 30min before ultraviolet irradiation.
3. The method for sterilizing by combining ultraviolet rays and sebacic acid according to claim 2, comprising the steps of:
(1) mixing the bacterial liquid with 1mM sebacic acid, placing the mixture in a six-hole plate, and setting ultraviolet irradiation conditions: wavelength range of 300-460nm, power of 18W, distance from the six-hole plate to the ultraviolet lamp tube of 8cm, and ultraviolet intensity of 2.4-3.0mW/cm2；
(2) Preheating the ultraviolet box for 30min, and placing the six-hole plate in the ultraviolet box for irradiating for 25-35 min.
4. The method for sterilizing by combining ultraviolet rays and sebacic acid according to claim 3, wherein the final concentration of the bacterial liquid in the step (1) is 106CFU/mL。
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011429762.5A CN112602707B (en) | 2020-12-09 | 2020-12-09 | Sebacic acid as ultraviolet sterilization synergist and application thereof |
| PCT/CN2021/118335 WO2022121420A1 (en) | 2020-12-09 | 2021-09-14 | Ultraviolet sterilization synergist and sterilization method using same in combination with ultraviolet |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011429762.5A CN112602707B (en) | 2020-12-09 | 2020-12-09 | Sebacic acid as ultraviolet sterilization synergist and application thereof |
Publications (2)
| Publication Number | Publication Date |
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