CN113842481B - Ultraviolet sterilization synergist lime essential oil and application thereof - Google Patents
Ultraviolet sterilization synergist lime essential oil and application thereof Download PDFInfo
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Classifications
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- 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
- A61L2/16—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using chemical substances
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Apparatus For Disinfection Or Sterilisation (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Food Preservation Except Freezing, Refrigeration, And Drying (AREA)
Abstract
The invention belongs to the technical field of ultraviolet sterilization, and discloses an ultraviolet sterilization synergist and an ultraviolet sterilization method combining the ultraviolet sterilization synergist. The research is based on that the Ultraviolet (UVA) and the lyme essential oil in the A wave band have weak sterilization effects (the sterilization amount is 0.5log-1.5log when the Ultraviolet (UVA) and the lyme essential oil act independently), and the Ultraviolet (UVA) and the lyme essential oil 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 ultraviolet sterilization synergist namely lyme essential oil 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 10nm to 400 nm. It is invisible light with a higher frequency than blue-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 becoming more serious 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 the sterilization technology of non-antibiotics is more and more applied in clinic, and the research on the ultraviolet sterilization effect is also becoming 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 high-efficiency, high-intensity and long-life ultraviolet rays are 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 uv alone is not sufficient to achieve a very good bactericidal effect, the sterilization time is long and the bacteria cannot be completely killed.
The lime essential oil is much lighter than the lemon essential oil, obviously has sweet smell, and belongs to citrus essential oil. It has effects in resisting ascorbic, bacteria and viruses, stimulating appetite, astringing, sterilizing, disinfecting, relieving fever, stopping bleeding, killing parasite, and promoting health. The essential oil obtained by squeezing the pericarp by a cold pressing method has chemical components and application range similar to those of the grapefruit essential oil. The antibacterial agent has the advantages of acquisition and certain antibacterial effect on bacteria. In order to enhance the ultraviolet sterilization effect, the research of the ultraviolet sterilization effect synergist is an essential step. However, the currently reported essential oil still has little synergistic effect on ultraviolet sterilization, and a new ultraviolet sterilization synergist needs to be supplemented to increase the sterilization effect of ultraviolet rays on various occasions.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provide the application of the lyme essential oil as the ultraviolet sterilization synergist.
The second purpose of the invention is to provide a sterilization method by using the lyme essential oil and ultraviolet.
The purpose of the invention is realized by the following technical scheme:
the application of the lyme essential oil as an ultraviolet sterilization synergist.
The research of the invention finds that the combination of the lyme essential oil and ultraviolet can obviously enhance the ultraviolet sterilization capability, so that the lyme essential oil can be used as a synergist for ultraviolet sterilization.
The invention also provides an ultraviolet sterilization synergist which comprises the lyme essential oil.
The invention also provides a method for sterilizing by combining ultraviolet with the lime essential oil, wherein the irradiation time of the ultraviolet is 30min, and the concentration of the lime essential oil is 1%.
Preferably, in the method for sterilizing, the ultraviolet intensity is 2.4-3.0mW/cm 2 。
Preferably, in the sterilization method, the preheating is performed for 30min before the ultraviolet irradiation.
Preferably, the sterilization method comprises the following steps:
(1) Mixing the bacterial liquid and 1% of lyme essential oil, placing the mixture into a six-hole plate, and setting ultraviolet irradiation conditions: wavelength range of 300-460nm, power of 18W, distance from six-hole plate to ultraviolet lamp tube of 8cm, ultraviolet intensity of 2.4-3.0mW/cm 2 ；
(2) Preheating the ultraviolet box for 20-40min, and placing the six-hole plate in the ultraviolet box for irradiating for 25-35min.
More preferably, the final concentration of the bacterial liquid in the step (1) of the sterilization method is 10 6 CFU/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 sterilizing effect of Ultraviolet (UVA) in the A wave band and lyme essential oil is weak (the sterilizing amount is 0.5log-1.5log when the ultraviolet and the lyme essential oil act independently), and the synergist has an obvious synergistic sterilizing effect 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.
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 is PHILIPS TL-D brand; wattage: 18W; voltage: 220V; wavelength: (UVA) 300-469nm; tube diameter of the lamp tube: 25mm; length: 60cm. The lyme essential oil is the Satya brand (italy).
A judgment principle: 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 essential oil treatment alone, and the essential oil and the ultraviolet have the synergistic sterilization effect.
Example 1 Sterilization Effect of different UV irradiation time on ATCC25922 Strain
1. Experimental materials:
(1) The ultraviolet lamp used for the test is a PHILIPS TL-D brand; wattage: 18W; voltage: 220V; wavelength: (UVA) 300-469nm; tube diameter of the lamp tube: 25mm; length: 60cm.
(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 55 ℃, 20mL of the MH agar medium and the MacConkey agar medium were put into a sterile petri dish by a pipette, and naturally dried for 30min to prepare the MH agar medium and the MacConkey agar medium.
Coli standard strain ATCC25922 (laboratory collection).
2. Preparation work before the test:
(1) Turning on 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 10 6 CFU/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 ATCC25922. 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 different concentrations of Lyme essential oil against ATCC25922 Strain
1. Experimental materials:
(1) And (3) testing: autoclaved MH broth was cooled for use.
(2) Resin azure: macklin (Merlin) brand, MW 251.17, purity 90%;
coli standard strain ATCC25922 (laboratory collection).
2. Preparation work before the test:
(1) Taking out the Lyme essential oil from the refrigerator, balancing to room temperature, and sucking part for later use;
(2) The E.coli standard strain ATCC25922 was cultured on MacconyKa agar medium to a suitable size.
(3) 0.2512g of resin azure powder is weighed into a centrifuge tube, 10mL of pure water is added for dissolution, and the concentration of the resin azure solution is 10mM/L.
3. Evaluation experiment of the bactericidal effect of the Lyme essential oil:
(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) Coli after incubation was diluted to 1 using MH broth00 times and about 10 times 6 CFU/mL for standby;
(3) Taking a sterile 96-well plate, adding 180 mu L of MH broth medium to the 1 st well, and adding 100 mu L of MH broth medium to the 2 nd-11 th well;
(4) Adding 20 mu L of essential oil with the original concentration into the 1 st hole, sucking 100 mu L to the 2 nd hole after blowing and beating uniformly, and repeating the steps, sucking 100 mu L from the 10 th hole and discarding; the concentration of the essential oil in each well is 10%, 5%, 2.5%, 1.25%, 0.625%, 0.3125%, 0.156%, 0.078%, 0.039%, 0.0195%.
(5) Adding 100 mu L of diluted bacteria liquid into the 1 st to 10 th holes, wherein the concentration of the essential oil in each hole is 5%, 2.5%, 1.25%, 0.625%, 0.3125%, 0.156%, 0.078%, 0.039%, 0.0195% and 0.01%, adding 100 mu L of bacteria liquid into the 11 th hole, and adding 200 mu L of LMH 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, sucking 10 mu L of 10mM/L resin azure with the concentration of 0.1mM/L, adding the resin azure into the hole, incubating for 2h, and reading the result.
The results are shown in Table 2, where the lime essential oil mic value is 0.313%, at which the growth of the bacteria is inhibited, and finally 1% lime essential oil concentration is selected as the use concentration of the UV enhancer.
TABLE 2 MIC of Lyme essential oil against ATCC25922 Strain and concentrations selected for the experiment
| Experimental strains | Lime essential oil Mic | Concentration for experiment |
| ATCC 25922 | 0.313％ | 1％ |
Example 3 bactericidal Effect of Lyme essential oil on ATCC25922 Strain after 30 minutes
1. Experimental materials:
(1) Test medium: an MH agar culture medium and a MacConKa agar culture medium (purchased from Guangdong Huanji Microscience Co., ltd.) which are sterilized under high pressure are cooled to 40 ℃, 20mL of the culture medium is taken into a sterile culture dish by a liquid transfer gun, and the culture medium is naturally dried for 30min to prepare the MH agar culture medium and the MacConKa agar culture medium.
Coli standard strain ATCC25922 (laboratory collection).
2. Preparation work before the test:
(1) Taking out the Lyme essential oil from the refrigerator, balancing to room temperature, and sucking part for later use;
(2) The E.coli standard strain ATCC25922 is cultured on a MacconKa medium to a suitable size.
3. Evaluation experiment of bactericidal effect of Lyme essential oil:
(1) Inoculating Escherichia coli ATCC25922, placing a 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 10 6 CFU/mL；
(3) Adding 1mL of bacterial liquid into a six-hole plate, adding 10 mu L of lyme essential oil with the original concentration, and uniformly mixing to ensure that the final concentration of the essential oil is 1%;
(4) Setting a control, processing a blank control and 1% of Lyme essential oil for 30min;
(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.
The results are shown in Table 3, the number of bacteria in the blank control group is not obviously different from the number of bacteria in the group which acts on the lime essential oil for 30min; it is demonstrated that 1% Lyme essential oil did not change the number of bacteria after 30min treatment with ATCC25922 bacteria. It can be seen that the effect of 1% of the lime essential oil alone has no obvious bacteriostatic effect on ATCC25922 bacteria.
TABLE 3 fungicidal Effect of Lyme essential oil on ATCC25922 strains
Example 4 evaluation of the killing effect of UV and UV boosters 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) 300-469nm; tube diameter of the lamp tube: 25mm; length: 60cm.
(2) Test medium: an MH agar culture medium and a MacConKa agar culture medium (purchased from Guangdong Huanji Microscience Co., ltd.) which are sterilized under high pressure are cooled to 40 ℃, 20mL of the culture medium is taken into a sterile culture dish by a liquid transfer gun, and the culture medium is naturally dried for 30min to prepare the MH agar culture medium and the MacConKa agar culture medium.
Coli standard strain ATCC25922 (laboratory collection).
2. Preparation work before testing:
(1) Turning on an ultraviolet lamp and continuously irradiating for 30 minutes for preheating;
(2) Taking out the lyme essential oil from the refrigerator, balancing to room temperature, and sucking a part for later use;
(3) The E.coli standard strain ATCC25922 was cultured on MacconyKa agar medium to a suitable size.
3. Ultraviolet synergist effect evaluation experiment:
(1) Inoculating Escherichia coli ATCC25922, placing the single colony in a centrifuge tube filled with 4mL MH broth, putting the centrifuge tube in a 37-degree shaking table, incubating at 180rpm for 4 hours, and taking out the centrifuge tube;
(2) Placing the centrifuge tube into a centrifuge for 5000 turnsAfter centrifugation for 8min, the supernatant is poured out and added with equal volume of normal saline for resuspension, and gradient dilution is carried out to ensure that the final bacterial load is 10 6 CFU/mL；
(3) Adding 1mL of bacterial liquid into a six-hole plate, adding 10 mu L of lyme essential oil with the original concentration, and uniformly mixing;
(4) Setting contrast, blank contrast, treating with 1% essential oil for shading, placing into ultraviolet irradiation box, and irradiating with ultraviolet for 30min;
(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% normal 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 37 ℃ is incubated for 16-18h for counting, and the statistical analysis is carried out after the experimental result is repeated by three biology.
In this experiment, a growth control group, an ultraviolet control group, an essential oil control group, and an ultraviolet plus essential oil test group were set, and the detection 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 1% lyme essential oil has no significant inhibitory effect on escherichia coli ATCC 25922; the combined action result of 30min shows that the synergistic effect of ultraviolet rays and 1% lyme essential oil has a remarkable sterilization effect on escherichia coli ATCC25922.
TABLE 4 germicidal Effect of UV and Lyme essential oil combinations on ATCC25922 strains
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 (5)
1. Use of Lyme essential oil as ultraviolet germicidal synergist, characterized in that the object to be sterilized is Escherichia coli ATCC25922.
2. A method for sterilizing by ultraviolet combined with Lyme essential oil is characterized in that the irradiation time of ultraviolet is 30min, the concentration of the Lyme essential oil is 1%, and the ultraviolet intensity is 2.4-3.0mW/cm 2 。
3. The method for sterilizing ultraviolet combined with Lyme essential oil according to claim 2, wherein the preheating is performed for 30min before the ultraviolet irradiation.
4. The method for sterilizing by ultraviolet combined with lyme essential oil according to claim 2, characterized by comprising the following steps:
(1) Mixing the bacterial liquid with 1% of Lyme essential oil, placing the mixture in a six-hole plate, and setting ultraviolet irradiation conditions: wavelength range of 300-460nm, power of 18W, distance from six-hole plate to ultraviolet lamp tube of 8cm, ultraviolet intensity of 2.4-3.0mW/cm 2 ；
(2) Preheating the ultraviolet box for 30min, and placing the six-hole plate in the ultraviolet box for irradiating for 25-35min.
5. The method for sterilizing by combining ultraviolet with Lyme essential oil according to claim 4, wherein the final concentration of the bacterial liquid in the step (1) is 10 6 CFU/mL。
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| ES2752448T5 (en) * | 2011-03-09 | 2022-08-26 | Marc Michael Baum | Formulations, methods and systems for straightening hair based on keratin |
| JP6135141B2 (en) * | 2013-01-18 | 2017-05-31 | 株式会社大林組 | UV sterilization method |
| IT201600109403A1 (en) * | 2016-10-28 | 2018-04-28 | Bio Lo Ga Srl | Composition for topical application with antifungal activity |
| CN112450229B (en) * | 2020-12-09 | 2021-06-15 | 华南农业大学 | Plant essential oil type ultraviolet sterilization synergist and ultraviolet combined sterilization method thereof |
| CN112602707B (en) * | 2020-12-09 | 2021-10-01 | 华南农业大学 | Sebacic acid as ultraviolet sterilization synergist and application thereof |
| CN112640923B (en) * | 2020-12-09 | 2021-07-02 | 华南农业大学 | Application of turmeric essential oil as ultraviolet sterilization synergist |
| CN112568243B (en) * | 2020-12-09 | 2021-10-08 | 华南农业大学 | Ultraviolet sterilization synergist and ultraviolet combined sterilization method thereof |
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