CN113994983B - Ultraviolet sterilization synergist grapefruit essential oil and application thereof - Google Patents
Ultraviolet sterilization synergist grapefruit essential oil and application thereof Download PDFInfo
- Publication number
- CN113994983B CN113994983B CN202111154366.0A CN202111154366A CN113994983B CN 113994983 B CN113994983 B CN 113994983B CN 202111154366 A CN202111154366 A CN 202111154366A CN 113994983 B CN113994983 B CN 113994983B
- Authority
- CN
- China
- Prior art keywords
- ultraviolet
- essential oil
- grapefruit essential
- sterilization
- grapefruit
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 239000000341 volatile oil Substances 0.000 title claims abstract description 56
- 230000001954 sterilising effect Effects 0.000 title claims abstract description 49
- 240000000560 Citrus x paradisi Species 0.000 title claims abstract description 47
- 238000004659 sterilization and disinfection Methods 0.000 title abstract description 39
- 238000000034 method Methods 0.000 claims abstract description 14
- 230000001580 bacterial effect Effects 0.000 claims description 23
- 239000007788 liquid Substances 0.000 claims description 18
- 230000001678 irradiating effect Effects 0.000 claims description 6
- 238000009281 ultraviolet germicidal irradiation Methods 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 2
- 230000000249 desinfective effect Effects 0.000 claims 2
- 241000894006 Bacteria Species 0.000 abstract description 20
- 230000000694 effects Effects 0.000 abstract description 15
- 238000005516 engineering process Methods 0.000 abstract description 7
- 230000002195 synergetic effect Effects 0.000 abstract description 6
- 238000011160 research Methods 0.000 abstract description 5
- 230000009471 action Effects 0.000 abstract description 4
- 239000002609 medium Substances 0.000 description 19
- 229920001817 Agar Polymers 0.000 description 15
- 239000008272 agar Substances 0.000 description 15
- 239000001963 growth medium Substances 0.000 description 11
- 238000012360 testing method Methods 0.000 description 11
- 241001360526 Escherichia coli ATCC 25922 Species 0.000 description 8
- 238000010790 dilution Methods 0.000 description 8
- 239000012895 dilution Substances 0.000 description 8
- 241000588724 Escherichia coli Species 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- 239000006154 MacConkey agar Substances 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 238000011156 evaluation Methods 0.000 description 5
- 239000002504 physiological saline solution Substances 0.000 description 5
- 230000005855 radiation Effects 0.000 description 5
- 230000002829 reductive effect Effects 0.000 description 5
- 239000011347 resin Substances 0.000 description 5
- 229920005989 resin Polymers 0.000 description 5
- 230000000844 anti-bacterial effect Effects 0.000 description 4
- 230000003385 bacteriostatic effect Effects 0.000 description 4
- 230000002070 germicidal effect Effects 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 206010059866 Drug resistance Diseases 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 238000007619 statistical method Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 102000053602 DNA Human genes 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- XMGQYMWWDOXHJM-UHFFFAOYSA-N limonene Chemical compound CC(=C)C1CCC(C)=CC1 XMGQYMWWDOXHJM-UHFFFAOYSA-N 0.000 description 2
- 229920002477 rna polymer Polymers 0.000 description 2
- 241000195493 Cryptophyta Species 0.000 description 1
- 241000272186 Falco columbarius Species 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 150000001299 aldehydes Chemical class 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
- 239000004599 antimicrobial Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 238000010009 beating Methods 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000011534 incubation Methods 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
- 230000002147 killing effect Effects 0.000 description 1
- 235000001510 limonene Nutrition 0.000 description 1
- 229940087305 limonene Drugs 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000001937 non-anti-biotic effect Effects 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 244000045947 parasite Species 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001172 regenerating effect Effects 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- 235000007586 terpenes Nutrition 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- 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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
-
- 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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/08—Magnoliopsida [dicotyledons]
- A01N65/36—Rutaceae [Rue family], e.g. lime, orange, lemon, corktree or pricklyash
-
- 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
-
- 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
- A61L2/18—Liquid substances or solutions comprising solids or dissolved gases
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Animal Behavior & Ethology (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Environmental Sciences (AREA)
- Agronomy & Crop Science (AREA)
- Zoology (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Wood Science & Technology (AREA)
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Botany (AREA)
- General Chemical & Material Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Apparatus For Disinfection Or Sterilisation (AREA)
Abstract
The invention belongs to the technical field of ultraviolet sterilization, and discloses an ultraviolet sterilization synergist and a method for sterilizing by combining ultraviolet sterilization synergist with ultraviolet. The research is based on that the A wave band Ultraviolet (UVA) and the grapefruit essential oil have weak sterilization effects (the sterilization amount is 0.5log-1.5log when the grapefruit essential oil is used alone), and the grapefruit essential oil and the UVA have obvious synergistic sterilization effects after combined action. Compared with the traditional ultraviolet sterilization technology, the ultraviolet sterilization technology 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 grapefruit 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 with a wavelength of 10nm to 400 nm. 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 cells of organisms, so that growing cells and/or regenerating cells are killed, and the sterilization effect is achieved. 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 that antibiotic resistance is not generated, 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.
The grapefruit essential oil is rich in terpene substances such as limonene and oxygen-containing compounds such as alcohols and aldehydes, which are extracted from grapefruit peel, and is widely applied to the fields of food, medical treatment, beauty treatment, daily use chemicals and the like in recent years due to unique fragrance, excellent quality and special functions. Furthermore, grapefruit essential oil is considered to be a promising preservative and antimicrobial agent. 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 grapefruit essential oil as the ultraviolet sterilization synergist.
The second purpose of the invention is to provide a method for sterilizing by combining grapefruit essential oil and ultraviolet.
The purpose of the invention is realized by the following technical scheme:
the grapefruit essential oil is used as an ultraviolet sterilization synergist.
The study of the invention finds that the combination of the grapefruit essential oil and ultraviolet can obviously enhance the ultraviolet sterilization capability, so that the grapefruit essential oil can be used as a synergist for ultraviolet sterilization.
The invention also provides an ultraviolet sterilization synergist which comprises grapefruit essential oil.
The invention also provides a method for sterilizing the grapefruit essential oil by combining ultraviolet radiation, wherein the ultraviolet radiation time is 30min, and the concentration of the grapefruit 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 grapefruit essential oil, placing the mixture in a six-hole plate, and setting ultraviolet irradiation conditions: the wavelength range is 300-460nm, the power is 18W, the distance from the six-hole plate to the ultraviolet lamp tube is 8cm, and the ultraviolet intensity is 2.4-3.0mW/cm 2 ；
(2) Preheating the ultraviolet box for 20-40min, and placing the six-hole plate in an 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 grapefruit essential oil is weak (the sterilizing amount is 0.5log-1.5log in the case of single action), and the synergist has an obvious synergistic sterilizing effect after combined action. Compared with the traditional ultraviolet sterilization technology, the ultraviolet sterilization technology 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 grapefruit essential oil is under the Satya brand (italy).
A judgment principle: for example, the bacterial amount reduced by the experimental group is reduced by 2 log values on the basis of the bacterial amount reduced by the ultraviolet treatment alone and the bacterial amount reduced by the essential oil treatment alone, namely, the essential oil and the ultraviolet have a synergistic sterilization 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 a PHILIPS TL-D brand; wattage: 18W; voltage: 220V; wavelength: (UVA) 300-469nm; tube diameter of the lamp tube: 25mm; length: 60cm.
(2) Culture medium for test: 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 is cultured on a MacconKa medium to a suitable size.
3. Ultraviolet sterilization effect evaluation experiment:
(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;
(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 37 ℃ is incubated for 16-18h for counting, and the statistical analysis is carried out after the experimental result is repeated by 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 grapefruit essential oil at various concentrations 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 testing:
(1) Taking the grapefruit essential oil out of the refrigerator, balancing to room temperature, and sucking a part for later use;
(2) The Escherichia coli standard strain ATCC25922 is cultured on an MacconKa agar medium to a proper 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 bactericidal effect of grapefruit 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) The incubated E.coli was diluted 100-fold to about 10-fold using MH broth 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 solution into the 1 st to 10 th holes, wherein the concentration of 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 solution into the 11 th hole, and adding 200 mu LMH broth into the 12 th hole;
(6) Repeating the steps (3) to (5) for three times to repeat in parallel;
(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.
As a result, as shown in Table 2, the grapefruit essential oil mic was 0.625%, at which the growth of bacteria was inhibited, and the grapefruit essential oil concentration of 1% was finally selected as the use concentration of the ultraviolet enhancer.
TABLE 2 MIC of grapefruit essential oil against ATCC25922 strain and concentration selected for experiments
| Experimental strains | Grapefruit essential oil Mic | Concentration for experiment |
| ATCC 25922 | 0.625％ | 1％ |
Example 3 Sterilization Effect of grapefruit essential oil on ATCC25922 Strain for 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 testing:
(1) Taking the grapefruit essential oil out of the refrigerator, balancing to room temperature, and sucking a part for later use;
(2) The E.coli standard strain ATCC25922 was cultured on MacconyKa medium to a suitable size.
3. Evaluation experiment of bactericidal effect of grapefruit 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 grapefruit 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 group and a 1% grapefruit essential oil group for 30min;
(5) And (3) absorbing 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 for gradient dilution, absorbing 25 mu L of diluted bacterial liquid, dripping the diluted bacterial liquid on an MH agar culture medium, incubating the diluted bacterial liquid in a 37-degree incubator for 16-18h, counting, and performing statistical analysis after three biological repetitions on experimental results.
The results are shown in Table 3, and the bacteria number of the blank control group and the bacteria number of the group which acts on the grapefruit essential oil for 30min are not obviously distinguished; it is demonstrated that 1% grapefruit essential oil showed no change in the number of bacteria after 30min treatment with ATCC25922 bacteria. It can be seen that 1% of grapefruit essential oil alone has no significant bacteriostatic effect on ATCC25922 bacteria.
TABLE 3 Bactericidal Effect of grapefruit essential oil 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 a PHILIPS TL-D brand; wattage: 18W; voltage: 220V; wavelength: (UVA) 300-469nm; tube diameter of the lamp tube: 25mm; length: 60cm.
(2) Culture medium for test: 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) Taking the grapefruit essential oil out of 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 in a centrifuge, centrifuging for 8min at 5000 rpm, pouring out the supernatant, adding equal volume of normal 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 grapefruit 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% 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.
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% grapefruit essential oil 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 1% of grapefruit essential oil has a remarkable sterilizing effect on escherichia coli ATCC 25922.
TABLE 4 Bactericidal Effect of combination of UV and grapefruit essential oil 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. A method for sterilizing grapefruit essential oil by combination of ultraviolet is characterized in that the irradiation time of ultraviolet is 25-35min, the concentration of grapefruit essential oil is 1%, and the ultraviolet intensity is 2.4-3.0mW/cm 2 。
2. A method of disinfecting grapefruit essential oil with UV combination according to claim 1, characterized in that the preheating is performed for 20-40min before the UV irradiation.
3. A method of disinfecting by ultraviolet combination grapefruit essential oil according to claim 2, characterized by comprising the steps of:
(1) Mixing the bacterial liquid with 1% of grapefruit 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 30min.
4. The method for sterilizing grapefruit essential oil in combination with ultraviolet rays as claimed in claim 3, wherein the final concentration of the bacterial liquid in the step (1) is 10 6 CFU/mL。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111154366.0A CN113994983B (en) | 2021-09-29 | 2021-09-29 | Ultraviolet sterilization synergist grapefruit essential oil and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111154366.0A CN113994983B (en) | 2021-09-29 | 2021-09-29 | Ultraviolet sterilization synergist grapefruit essential oil and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113994983A CN113994983A (en) | 2022-02-01 |
| CN113994983B true CN113994983B (en) | 2022-10-28 |
Family
ID=79922087
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111154366.0A Active CN113994983B (en) | 2021-09-29 | 2021-09-29 | Ultraviolet sterilization synergist grapefruit essential oil and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113994983B (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006109898A1 (en) * | 2005-04-09 | 2006-10-19 | Pion-Tech Co., Ltd | Antimicrobial composition containing natural extract, nano silver and natural essential oil |
| CN104338168A (en) * | 2013-08-02 | 2015-02-11 | 雷兆凤 | Air freshener for upper respiratory infection patients and preparing method thereof |
| CN105688245A (en) * | 2014-11-28 | 2016-06-22 | 青岛首冠企业管理咨询有限公司 | Air freshener |
| CN108935456A (en) * | 2018-07-06 | 2018-12-07 | 青岛日晟源作物营养有限公司 | A kind of pesticide synergistic agent and its application in Herbicidal combinations, Pesticidal combination and bactericidal composition |
| CN112568243A (en) * | 2020-12-09 | 2021-03-30 | 华南农业大学 | Ultraviolet sterilization synergist and ultraviolet combined sterilization method thereof |
| CN112640923A (en) * | 2020-12-09 | 2021-04-13 | 华南农业大学 | Application of turmeric essential oil as ultraviolet sterilization synergist |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111480647A (en) * | 2020-05-14 | 2020-08-04 | 上海赛境实业有限公司 | Bactericidal natural plant essential oil aromatherapy and preparation method and application thereof |
-
2021
- 2021-09-29 CN CN202111154366.0A patent/CN113994983B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006109898A1 (en) * | 2005-04-09 | 2006-10-19 | Pion-Tech Co., Ltd | Antimicrobial composition containing natural extract, nano silver and natural essential oil |
| CN104338168A (en) * | 2013-08-02 | 2015-02-11 | 雷兆凤 | Air freshener for upper respiratory infection patients and preparing method thereof |
| CN105688245A (en) * | 2014-11-28 | 2016-06-22 | 青岛首冠企业管理咨询有限公司 | Air freshener |
| CN108935456A (en) * | 2018-07-06 | 2018-12-07 | 青岛日晟源作物营养有限公司 | A kind of pesticide synergistic agent and its application in Herbicidal combinations, Pesticidal combination and bactericidal composition |
| CN112568243A (en) * | 2020-12-09 | 2021-03-30 | 华南农业大学 | Ultraviolet sterilization synergist and ultraviolet combined sterilization method thereof |
| CN112640923A (en) * | 2020-12-09 | 2021-04-13 | 华南农业大学 | Application of turmeric essential oil as ultraviolet sterilization synergist |
Non-Patent Citations (2)
| Title |
|---|
| LED紫外线灯杀菌效果试验研究;夏冠英炫等;《中国消毒学杂志》;20170115;第34卷(第01期);第1-2页 * |
| 葡萄柚精油抑菌活性的研究;李悦 等;《食品研究与开发》;20101130;第31卷(第11期);摘要、正文第1.3.1节 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113994983A (en) | 2022-02-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112640923B (en) | Application of turmeric essential oil as ultraviolet sterilization synergist | |
| CN112568243B (en) | Ultraviolet sterilization synergist and ultraviolet combined sterilization method thereof | |
| Rifna et al. | Emerging technology applications for improving seed germination | |
| Oguma et al. | Photoreactivation of Legionella pneumophila after inactivation by low-or medium-pressure ultraviolet lamp | |
| Najdovski et al. | The killing activity of microwaves on some non-sporogenic and sporogenic medically important bacterial strains | |
| CN112450229B (en) | Plant essential oil type ultraviolet sterilization synergist and ultraviolet combined sterilization method thereof | |
| CN111803631B (en) | Preparation method and application of carbon nanodots with efficient antibacterial property | |
| CN112602707B (en) | Sebacic acid as ultraviolet sterilization synergist and application thereof | |
| US20230321288A1 (en) | Riboflavin photochemical treatment (rpt)-based inactivation method of pathogens in biological liquid sample | |
| CN113994983B (en) | Ultraviolet sterilization synergist grapefruit essential oil and application thereof | |
| CN112655708A (en) | Ultraviolet sterilization synergist (S) -2-hydroxy-4-methyl valeric acid and application thereof | |
| CN113842481B (en) | Ultraviolet sterilization synergist lime essential oil and application thereof | |
| CN113907094B (en) | Ultraviolet sterilization synergist-France cypress essential oil and application thereof | |
| Maclean et al. | Photoinactivation and photoreactivation responses by bacterial pathogens after exposure to pulsed UV-light | |
| WO2022121420A1 (en) | Ultraviolet sterilization synergist and sterilization method using same in combination with ultraviolet | |
| Shareef et al. | Sterilization of culture media for microorganisms using a microwave oven instead of autoclave | |
| CN104984377A (en) | Multiband LED ultraviolet disinfection apparatus | |
| CN112624455A (en) | Method for disinfecting gram-negative superbacteria with antibiotic resistance in water | |
| CN111841599A (en) | Carbon quantum dot doped carbon nitride composite nano material with photocatalytic antibacterial performance and preparation method and application thereof | |
| CN206298475U (en) | One kind production urea for vehicle ultraviolet sterilization apparatus | |
| Thiyagarajan et al. | Portable plasma medical device for infection treatment | |
| CN1295972C (en) | Application of ultraviolet light sterilization technology in processing Pu'er tea | |
| CN212817295U (en) | LED disinfection lamp that disinfects based on UVC and UVA dual wavelength gain | |
| CN114891346B (en) | Polylysine compound based on molybdenum polyacid and application thereof in antibacterial field | |
| CN117379574B (en) | Photodynamic sterilization method using epsilon-polylysine combined with curcumin as mediation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |