CN113068791B - Method for improving sterilization efficiency by combining photodynamic technology with alkaline electrolyzed water - Google Patents
Method for improving sterilization efficiency by combining photodynamic technology with alkaline electrolyzed water Download PDFInfo
- Publication number
- CN113068791B CN113068791B CN202110382052.XA CN202110382052A CN113068791B CN 113068791 B CN113068791 B CN 113068791B CN 202110382052 A CN202110382052 A CN 202110382052A CN 113068791 B CN113068791 B CN 113068791B
- Authority
- CN
- China
- Prior art keywords
- electrolyzed water
- photosensitizer
- alkaline electrolyzed
- sterilization efficiency
- water
- 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
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 113
- 230000001954 sterilising effect Effects 0.000 title claims abstract description 59
- 238000004659 sterilization and disinfection Methods 0.000 title claims abstract description 59
- 238000000034 method Methods 0.000 title claims abstract description 49
- 238000005516 engineering process Methods 0.000 title claims abstract description 41
- 239000003504 photosensitizing agent Substances 0.000 claims abstract description 68
- 239000006185 dispersion Substances 0.000 claims abstract description 11
- 241000607272 Vibrio parahaemolyticus Species 0.000 claims abstract description 9
- 241000863430 Shewanella Species 0.000 claims abstract description 7
- 238000002156 mixing Methods 0.000 claims abstract description 7
- 230000001678 irradiating effect Effects 0.000 claims abstract description 4
- 239000007788 liquid Substances 0.000 claims abstract description 4
- HWDGVJUIHRPKFR-UHFFFAOYSA-I copper;trisodium;18-(2-carboxylatoethyl)-20-(carboxylatomethyl)-12-ethenyl-7-ethyl-3,8,13,17-tetramethyl-17,18-dihydroporphyrin-21,23-diide-2-carboxylate Chemical group [Na+].[Na+].[Na+].[Cu+2].N1=C(C(CC([O-])=O)=C2C(C(C)C(C=C3C(=C(C=C)C(=C4)[N-]3)C)=N2)CCC([O-])=O)C(=C([O-])[O-])C(C)=C1C=C1C(CC)=C(C)C4=N1 HWDGVJUIHRPKFR-UHFFFAOYSA-I 0.000 claims description 18
- 229940079841 sodium copper chlorophyllin Drugs 0.000 claims description 18
- 235000013758 sodium copper chlorophyllin Nutrition 0.000 claims description 18
- 241000894006 Bacteria Species 0.000 claims description 13
- 238000002360 preparation method Methods 0.000 claims description 10
- 238000011534 incubation Methods 0.000 claims description 8
- 239000008399 tap water Substances 0.000 claims description 4
- 235000020679 tap water Nutrition 0.000 claims description 4
- 230000006872 improvement Effects 0.000 claims description 3
- 230000033116 oxidation-reduction process Effects 0.000 claims description 3
- 239000003513 alkali Substances 0.000 claims 1
- 230000002829 reductive effect Effects 0.000 abstract description 23
- 244000005700 microbiome Species 0.000 abstract description 9
- 238000005868 electrolysis reaction Methods 0.000 abstract description 6
- 230000008901 benefit Effects 0.000 abstract description 5
- 238000002428 photodynamic therapy Methods 0.000 description 63
- 230000000694 effects Effects 0.000 description 26
- 239000000243 solution Substances 0.000 description 25
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 24
- 238000005286 illumination Methods 0.000 description 23
- 241000894007 species Species 0.000 description 22
- 239000002028 Biomass Substances 0.000 description 17
- 241000251468 Actinopterygii Species 0.000 description 12
- 239000011780 sodium chloride Substances 0.000 description 12
- 229910001220 stainless steel Inorganic materials 0.000 description 11
- 239000010935 stainless steel Substances 0.000 description 11
- 238000002835 absorbance Methods 0.000 description 10
- 239000013078 crystal Substances 0.000 description 10
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 9
- 239000000523 sample Substances 0.000 description 9
- 230000003833 cell viability Effects 0.000 description 8
- 239000000203 mixture Substances 0.000 description 8
- 239000002105 nanoparticle Substances 0.000 description 8
- 239000003153 chemical reaction reagent Substances 0.000 description 7
- 239000013642 negative control Substances 0.000 description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 6
- 150000001875 compounds Chemical class 0.000 description 6
- 238000004140 cleaning Methods 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- GEICAQNIOJFRQN-UHFFFAOYSA-N 9-aminomethyl-9,10-dihydroanthracene Chemical compound C1=CC=C2C(CN)C3=CC=CC=C3CC2=C1 GEICAQNIOJFRQN-UHFFFAOYSA-N 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 4
- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 description 4
- 239000008272 agar Substances 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 230000002779 inactivation Effects 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 230000002035 prolonged effect Effects 0.000 description 4
- 239000003642 reactive oxygen metabolite Substances 0.000 description 4
- 230000003068 static effect Effects 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 230000003247 decreasing effect Effects 0.000 description 3
- 229910052742 iron Inorganic materials 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 239000011148 porous material Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- AUNGANRZJHBGPY-UHFFFAOYSA-N D-Lyxoflavin Natural products OCC(O)C(O)C(O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 239000004793 Polystyrene Substances 0.000 description 2
- 241000194019 Streptococcus mutans Species 0.000 description 2
- 241000953555 Theama Species 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 229930002875 chlorophyll Natural products 0.000 description 2
- 235000019804 chlorophyll Nutrition 0.000 description 2
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 description 2
- 229940099898 chlorophyllin Drugs 0.000 description 2
- 235000019805 chlorophyllin Nutrition 0.000 description 2
- VFLDPWHFBUODDF-FCXRPNKRSA-N curcumin Chemical compound C1=C(O)C(OC)=CC(\C=C\C(=O)CC(=O)\C=C\C=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-FCXRPNKRSA-N 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- 231100000956 nontoxicity Toxicity 0.000 description 2
- 239000004033 plastic Substances 0.000 description 2
- 229920003023 plastic Polymers 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 229920002223 polystyrene Polymers 0.000 description 2
- 229960002477 riboflavin Drugs 0.000 description 2
- 239000002151 riboflavin Substances 0.000 description 2
- 235000019192 riboflavin Nutrition 0.000 description 2
- 229910052709 silver Inorganic materials 0.000 description 2
- 239000004332 silver Substances 0.000 description 2
- -1 superoxide anions Chemical class 0.000 description 2
- 230000008685 targeting Effects 0.000 description 2
- 229950003937 tolonium Drugs 0.000 description 2
- HNONEKILPDHFOL-UHFFFAOYSA-M tolonium chloride Chemical compound [Cl-].C1=C(C)C(N)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 HNONEKILPDHFOL-UHFFFAOYSA-M 0.000 description 2
- BBDHEBFBEPYAMC-UHFFFAOYSA-N 2-[10-(carboxymethyl)anthracen-9-yl]acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=C(C=CC=C3)C3=C(CC(O)=O)C2=C1 BBDHEBFBEPYAMC-UHFFFAOYSA-N 0.000 description 1
- 239000010963 304 stainless steel Substances 0.000 description 1
- 241000472349 Asparagus adscendens Species 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 241000238557 Decapoda Species 0.000 description 1
- 235000017274 Diospyros sandwicensis Nutrition 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000282838 Lama Species 0.000 description 1
- 102000006833 Multifunctional Enzymes Human genes 0.000 description 1
- 108010047290 Multifunctional Enzymes Proteins 0.000 description 1
- 229910000589 SAE 304 stainless steel Inorganic materials 0.000 description 1
- 241000607142 Salmonella Species 0.000 description 1
- 241001138501 Salmonella enterica Species 0.000 description 1
- 241000863432 Shewanella putrefaciens Species 0.000 description 1
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000003214 anti-biofilm Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- SWJRLTOYMKAFKN-UHFFFAOYSA-N azanium 2-(2,5-diphenyl-1H-tetrazol-1-ium-3-yl)-4,5-dimethyl-1,3-thiazole dibromide Chemical compound [Br-].[NH4+].CC=1N=C(SC1C)N1N([NH2+]C(=N1)C1=CC=CC=C1)C1=CC=CC=C1.[Br-] SWJRLTOYMKAFKN-UHFFFAOYSA-N 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000000941 bile Anatomy 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- HWDGVJUIHRPKFR-ZWPRWVNUSA-I copper;trisodium;3-[(2s,3s)-20-(carboxylatomethyl)-18-(dioxidomethylidene)-8-ethenyl-13-ethyl-3,7,12,17-tetramethyl-2,3-dihydroporphyrin-23-id-2-yl]propanoate Chemical compound [Na+].[Na+].[Na+].[Cu+2].C1=C([N-]2)C(CC)=C(C)C2=CC(C(=C2C)C=C)=NC2=CC([C@H]([C@@H]2CCC([O-])=O)C)=NC2=C(CC([O-])=O)C2=NC1=C(C)C2=C([O-])[O-] HWDGVJUIHRPKFR-ZWPRWVNUSA-I 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 239000004148 curcumin Substances 0.000 description 1
- 235000012754 curcumin Nutrition 0.000 description 1
- 229940109262 curcumin Drugs 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000002498 deadly effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- VFLDPWHFBUODDF-UHFFFAOYSA-N diferuloylmethane Natural products C1=C(O)C(OC)=CC(C=CC(=O)CC(=O)C=CC=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-UHFFFAOYSA-N 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000005684 electric field Effects 0.000 description 1
- 239000008151 electrolyte solution Substances 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000000556 factor analysis Methods 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- NEMFQSKAPLGFIP-UHFFFAOYSA-N magnesiosodium Chemical compound [Na].[Mg] NEMFQSKAPLGFIP-UHFFFAOYSA-N 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000002715 modification method Methods 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 230000000886 photobiology Effects 0.000 description 1
- 208000017983 photosensitivity disease Diseases 0.000 description 1
- 231100000434 photosensitization Toxicity 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- DHCDFWKWKRSZHF-UHFFFAOYSA-L thiosulfate(2-) Chemical compound [O-]S([S-])(=O)=O DHCDFWKWKRSZHF-UHFFFAOYSA-L 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/27—Removal of unwanted matter, e.g. deodorisation or detoxification by chemical treatment, by adsorption or by absorption
- A23L5/276—Treatment with inorganic compounds
Landscapes
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Apparatus For Disinfection Or Sterilisation (AREA)
Abstract
The invention discloses a method for improving sterilization efficiency by combining a photodynamic technology with alkaline electrolyzed water, which comprises the steps of mixing a photosensitizer dispersion liquid with a sample to be treated, wherein the photosensitizer dispersion liquid comprises a photosensitizer and the alkaline electrolyzed water; incubating the mixed sample; and irradiating the incubated sample with a light source. The method creatively combines the photodynamic technology with the water electrolysis technology, and opens up a brand new channel for improving the PDT sterilization efficiency; the technology can be applied to the food industry by matching alkaline electrolyzed water with a food-grade photosensitizer, so that the risk brought by pollution of harmful microorganisms is reduced; the method has the advantages of simple operation, good applicability, low cost, good safety, green and pollution-free performance, and capability of remarkably improving the sterilization efficiency (on vibrio parahaemolyticus and Shewanella putrefying) and great application prospect.
Description
Technical Field
The invention belongs to the field of food safety and biotechnology, and relates to a method for improving sterilization efficiency by combining a photodynamic technology with alkaline electrolyzed water.
Background
Photodynamic technology (photodynamic technology, PDT), also known as photodynamic inactivation (photodynamic inactivation, PDI), is a new method of inactivating tumor cells and pathogenic microorganisms (including bacteria, fungi and viruses) that is widely used in clinical medicine. Three requirements for PDT sterilization are: oxygen, light and photosensitizers. The three combine to produce reactive oxygen species (reactive oxygen species, ROS), including singlet oxygen, hydrogen peroxide, hydroxyl radicals, and superoxide anions. ROS have a strong oxidizing effect, thereby damaging or dying cells. The technology has the advantages of low cost, green safety, no toxicity, no pollution and the like, and has been applied to the field of food safety in recent years. However, PDT also has some significant drawbacks: the sterilization efficiency to some harmful microorganisms and the biological film thereof is not high, or the selected photosensitizer has better antibacterial effect but has toxicity and is difficult to be applied to the food industry.
The most common method for improving the PDT sterilization efficiency is the photosensitizer modification method. Specifically, a photosensitizer is combined with nanoparticles or some compounds and derivatives thereof. For example, some nano particles (silver nano or gold nano and the like) can be used as carriers for conveying the photosensitizer to a target for targeting, and the nano particles have good stability, strong targeting and large specific surface area, can carry more photosensitizer molecules to approach or enter the inside of cells, and further improve the sterilization efficiency. Some compounds and photosensitizers are combined to improve the sterilization efficiency by directly or indirectly changing the molecular structure of the photosensitizers by utilizing the special properties of the compounds, improving the solubility, the combination rate with bacteria, the self stability and the like. As in literature one (Misba Lama, kulshrestha Shatavari, khan Asad U.S. anti-biochem action of a toluidine blue O-silver nanoparticle conjugate on Streptococcus mutans: a mechanism of type I photodynamic therapy [ J ]. Biofouling,2016,32 (3): 313-328.), it is pointed out that toluidine blue-mediated photodynamic technology in combination with nanoparticles enhances the anti-biofilm effect on Streptococcus mutans. Although such methods improve the sterilization efficiency to some extent, the cost of the nanoparticles or compounds is high, the synthesis process is complex, and the nanoparticles or compounds are more deadly often toxic, which greatly reduces the application range of the nanoparticles or compounds in the food safety field.
Yet another common approach is to combine PDT with other non-thermal sterilization techniques (ultrasound, ultraviolet, high voltage pulsed electric fields, etc.). PDT has poor biofilm removal because the inclusion of thick extracellular polymers outside the biofilm can block photosensitizer molecules from entering the bacteria. When PDT and ultrasonic waves are combined to treat the biological envelope, the extracellular polymer can be destroyed by the ultrasonic action, and the binding rate of the photosensitizer and the target is improved, so that the sterilization efficiency is enhanced. As in document two (Buchotec Irina, lukseviciute Viktorija, kokstaite Rita, et al, inactive of Gram (-) bacteria Salmonella enterica by chlorophyllin-based photosensitization: mechanism of action and new strategies to enhance the inactivation efficiency [ J ]. Journal of Photochemistry and Photobiology B-Biology,2017,172,1-10.), chlorophyll-mediated photodynamic combined with high-power pulsed ultraviolet light technology has been shown to significantly improve the efficiency of salmonella inactivation. Although the sterilization efficiency is improved to a certain extent by the method, the cost is too high, the process is complex, and each technology has a certain application range, so that the more the technologies are used in combination, the smaller the application range is.
Therefore, the development of a method for improving the PDT sterilization efficiency, which has low cost and good applicability, has very practical significance.
Disclosure of Invention
The invention aims to overcome the defects that the existing method for improving the PDT sterilization efficiency is high in cost, complex in process, small in application range, incapable of efficiently removing the biological film of a mixture species with stronger resistance in the food industry and the like, and provides a method for improving the PDT sterilization efficiency, which is low in cost and good in applicability, for food safety. The invention creatively combines the photodynamic technology with the water electrolysis technology, opens up a brand-new channel for improving the PDT sterilization efficiency, and has great application potential in the field of future food safety prevention and control.
In order to achieve the above purpose, the present invention provides the following technical solutions:
a method for improving sterilization efficiency by combining photodynamic technology with alkaline electrolyzed water comprises the following steps:
(1) Mixing a photosensitizer dispersion with a sample to be treated, wherein the photosensitizer dispersion comprises a photosensitizer and alkaline electrolyzed water;
(2) Incubating the mixed sample;
(3) And irradiating the incubated sample with a light source.
The invention improves the existing PDT sterilization technology by using alkaline electrolyzed water to improve sterilization efficiency, wherein the alkaline electrolyzed water is water with special properties, electrolyte solution passes through an electrolysis device, and is electrolyzed water with reducibility generated at the cathode of an electrolysis tank, compared with common tap water, the bond length of H-O ionic bonds in the water is increased, the bond angle is increased, the attractive force between water molecules is reduced, and the surface tension of the water is reduced, so that the alkaline electrolyzed water has very high activity; meanwhile, due to the electrolysis, the solution has an alkaline environment, contains a large amount of free charge ions and functional groups, has good rust resistance, good oxidation resistance and high permeability. The source of the microbial agent is green, safe, nontoxic and harmless, and the microbial agent can be matched with the microbial agent to realize remarkable improvement of sterilization efficiency. The technology is expected to be applied to the food industry, so that the risk brought by pollution of harmful microorganisms is reduced, and the technology has a great application prospect.
As a preferable technical scheme:
the method for improving the sterilization efficiency by combining the photodynamic technology with alkaline electrolyzed water is characterized in that the photosensitizer is selected from more than one of sodium copper chlorophyllin and riboflavin, the invention is applicable to all water-soluble photosensitizers such as sodium magnesium chlorophyllin, riboflavin and the like or a mixture of the photosensitizers (the premise of adopting mixed photosensitizers is that the mixed photosensitizers do not react with each other and the maximum absorption wavelength is close), other water-insoluble photosensitizers such as curcumin and the like are also applicable, and the effect is slightly poorer than that of the water-soluble photosensitizers due to poor solubility. The sodium copper chlorophyllin is water-soluble, has wide sources, low price, no toxicity and no pollution, is used as a food additive in multiple countries, namely, is of a food safety level, and can be applied to food sterilization to reduce risks caused by pollution of harmful microorganisms.
The method for improving the sterilization efficiency by combining the photodynamic technology with the alkaline electrolyzed water comprises the following steps of:
preparing alkaline electrolyzed water by using an electrolyzed water preparation instrument, wherein a water inlet is common tap water, a water outlet is target electrolyzed water, and parameters of the electrolyzed water preparation instrument are as follows: ph=7 to 10, oxidation-reduction potential (ORP) =0 to-400 mV (preferably, ph=9 to 10, orp= -300 to-400 mV, more preferably, ph=9.80, orp= -388 mV). Wherein, the pH value is inversely proportional to the ORP value, and the higher the pH value is, the better the sterilization efficiency is, but the pH value is not too high, because the food quality and even the human health are damaged. The alkaline electrolyzed water adopted by the invention is novel environment-friendly water which is green, safe, nontoxic and harmless.
The method for improving the sterilization efficiency by combining the photodynamic technology with alkaline electrolyzed water, wherein the concentration of the photosensitizer in a sample mixing system to be treated is 0-1000 mu M, preferably the concentration of the photosensitizer in the sample mixing system to be treated is 80-1000 mu M, more preferably the concentration of the photosensitizer in the sample mixing system to be treated is 150-300 mu M, has the advantages that the sterilization efficiency is gradually improved along with the increase of the concentration of the photosensitizer, the maximum sterilization effect is achieved when the concentration of the photosensitizer in the sample mixing system to be treated is increased to 150 mu M, and the sterilization efficiency is not improved obviously after the concentration of the photosensitizer is continuously increased, so that the optimal concentration of the photosensitizer is 150 mu M. Of course, the protection scope of the present invention is not limited to this, and those skilled in the art can adjust the addition amount of the photosensitizer according to the actual requirement in practical application, and only one possible technical scheme is given here.
In the method for improving the sterilization efficiency by combining the photodynamic technology with alkaline electrolyzed water, in the step (2), the incubation is performed under a dark condition for 0-60 min, preferably for 10-60 min, and more preferably for 20min. In addition, the incubation temperature has no influence on the invention, and the temperature of 0-40 ℃ can normally work, and the too low or too high temperature can possibly influence the normal use of the photosensitizer and the illumination device, and the invention is carried out at room temperature, namely 25 ℃.
In the method for improving the sterilization efficiency by combining the photodynamic technology with alkaline electrolyzed water, in the step (3), the light source is a blue light source with the wavelength of 455-460 nm. Only one possible technical solution is given here, and in practical application, a person skilled in the art can adjust relevant conditions such as the light source according to actual requirements.
The method for improving the sterilization efficiency by combining the photodynamic technology with alkaline electrolyzed water comprises the following steps of (3) irradiating the light source with the light power density of 0-500 mW/cm 2 Preferably, the light source irradiates light with a power density of 2-50 mW/cm 2 More preferably, the invention selects 3.8mW/cm 2 The optical power density of the (C) is extremely low in energy consumption and has a remarkable sterilization effect. Of course, the protection scope of the present invention is not limited to this, and a person skilled in the art can adjust the optical power density according to the actual requirement in practical application, only one possible technical scheme is given here, and theoretically, the higher the power, the better the sterilization effect. In addition, the invention is not limited to blue light source, and any light source and optical power density can be used for enabling the photosensitizer to excite and generate ROS to generate photodynamic reaction, and the alkaline electrolyzed water can be combined with the light source to improve the sterilization efficiency. The irradiation time of the light source is 0-120 min. Preferably, the illumination time of the light source is 10-120 min, more preferably, the illumination time is 20-120 min, and experiments show that the sterilization efficiency is gradually improved along with the extension of the illumination time, and the maximum sterilization effect is achieved when the illumination time is prolonged to 20min, and the sterilization efficiency is not obviously improved after the illumination time is prolonged, so that the optimal illumination time is 20min.
The method for improving the sterilization efficiency by combining the photodynamic technology with alkaline electrolyzed water is used for improving the sterilization efficiency of vibrio parahaemolyticus (pathogenic bacteria) and Shewanella putrefying (putrefying bacteria). The protective scope of the invention is not limited thereto, and only a portion of the species for which the specific tests herein are effective are presented, the method of the invention may also be applied to killing a variety of other common pathogenic and spoilage bacteria.
In addition, the invention also provides application of the method in sterilization.
The invention also provides a photosensitizer composition (corresponding to the photosensitizer dispersion) which comprises a photosensitizer and alkaline electrolyzed water.
The beneficial effects are that:
(1) The invention combines the photodynamic technology with the alkaline electrolyzed water to improve the sterilization efficiency, creatively combines the photodynamic technology with the electrolyzed water technology, and opens up a brand new channel for improving the PDT sterilization efficiency;
(2) The photodynamic technology is combined with the alkaline electrolyzed water to improve the sterilization efficiency, and the alkaline electrolyzed water is matched with the food-grade photosensitizer, so that the technology can be applied to the food industry, and the risk brought by pollution of harmful microorganisms is reduced;
(3) The method for improving the sterilization efficiency by combining the photodynamic technology with alkaline electrolyzed water has the advantages of extremely simple operation, good applicability, low cost, good safety, green and pollution-free performance, can obviously remove the biofilm, can obviously improve the sterilization efficiency (on vibrio parahaemolyticus and Shewanella putrefying) and has great application prospect.
Drawings
FIG. 1 is a schematic view of the device structure of a 455-460 nm LED lighting system used in the embodiments;
FIG. 2 is a graph showing comparison of ROS and singlet oxygen expression levels under various concentrations of sodium copper chlorophyllin (FIGS. 2-A, 2-B) and light exposure time (FIGS. 2-C, 2-D) treatment conditions of PDT in combination with alkaline electrolyzed water;
FIG. 3 is a graph showing the effect of PDT in combination with alkaline electrolyzed water on total biofilm biomass of a mixture of species under treatment conditions of varying concentrations of sodium copper chlorophyllin (FIG. 3-A) and light exposure time (FIG. 3-B);
FIG. 4 is a graph showing the effect of PDT in combination with alkaline electrolyzed water treatment on biofilm cell viability (FIG. 4-A) and total colony count (FIG. 4-B) of a mixed species;
FIG. 5 is a graph showing the effect of PDT in combination with alkaline electrolyzed water treatment on total biofilm biomass and cell viability of a mixture of species on the surface of stainless steel (FIGS. 5-A, 5-C) and fish scales (FIGS. 5-B, 5-D);
wherein, 1-LED shoots the lamp house, 2-elevating platform, 3-24 orifice plates, 4-LED blue light source.
Detailed Description
The following detailed description of the invention will be further presented in conjunction with the appended drawings, and it will be apparent that the described embodiments are merely some, but not all, examples of the invention.
The strains used in the examples were as follows:
vibrio parahaemolyticus (Vibrio parahaemolyticus, VP) ATCC 17802 is from the institute of microorganisms of the national academy of sciences; shewanella putrefying (Shewanella putrefaciens, SP) strain E05 was isolated from prawns by the present laboratory.
The reagents and materials used in the examples are as follows:
sodium copper chlorophyllin (sodium copper chlorophyllin, food grade, purity > 95%) Shanghai bioengineering, inc.; alkaline electrolyzed water (ph=9.80, orp= -388 mV) was prepared from an electrolyzed water preparation apparatus; thiosulfate citrate bile sucrose agar medium (TCBS), tryptone soy broth medium (TSB), iron agar medium, beijing land bridge technologies inc; ROS reactive oxygen species fluorescent probe Beijing Soy Biotechnology Co., ltd; 9,10 anthracenediyl-bis (methylene) dicarboxylic acid (AMDA, 10 mM) Sigma, america; 3- (4, 5-dimethylthiazol-2-yl) -2, 5-diphenyltetrazolium ammonium bromide (MTT reagent, 5 mg/mL), PBS phosphate buffer solution, crystal Violet solution Shanghai Biotechnology Co., ltd; other chemical analysis reagents were purchased from national drug group chemical reagent limited. 304 stainless steel discs (d=14 mm) and fresh round fish scales (d=14 mm) were purchased in the local understory market.
The instruments used in the examples were as follows:
the 455-460 nm LED lighting system is shown in figure 1, and comprises an LED shooting lamp box 1, a lifting table 2, a 24 pore plate 3 and an LED blue light source 4, wherein the LED lighting system is surrounded by the LED shooting lamp box and can prevent external light from entering, the LED blue light source is arranged on the inner side of the top of the LED shooting lamp box, the 24 pore plate is arranged on the lifting table, the distance between the LED blue light source and a sample in the 24 pore plate is adjusted to 5.0cm by the lifting table, the LED blue light source (wavelength 455-460 nm, the length of a lamp tube is 30cm, and the irradiation dose E=3.80 mJ/c per secondm 2 ) Purchased from philips lighting electronics, inc., shanghai;
full-automatic bench centrifuge 5417R, available from Eppendorf, germany;
the multifunctional enzyme-labeled instrument, synergy htx, was purchased from berteng instruments ltd;
electrolyzed water forming apparatus FW-200, available from Amano corporation, japan.
Method for culturing hybrid biofilm (i.e., method for preparing sample to be treated)
Streaking Vibrio parahaemolyticus (-80 ℃ C. Stored in 25% glycerol tube) on TCBS plate, culturing at 37 ℃ C. For 12h, picking up green single colony, placing into TSB (containing 3% NaCl) broth, and culturing overnight (200 rpm) in a constant temperature (37 ℃) incubator; shewanella putrefying (-80 ℃ C. Stored in 25% glycerol tube) was streaked onto iron agar plates, cultured at 25 ℃ C. For 24h, and then black single colonies were picked up in TSB broth and placed in a constant temperature (30 ℃) incubator overnight (200 rpm). The concentration of the two bacteria is respectively regulated to 8log CFU/mL by using 0.85% NaCl solution, and then the bacteria solutions of the two bacteria are mixed in equal volume for standby.
10. Mu.L of the above mixed bacterial solution and 990. Mu.L of sterile fresh TSB medium were added to a 24-well polystyrene plate, and the plate was then placed in a 25℃incubator for 18h of stationary culture to form a mature mixed species biofilm.
Preparation of photosensitizer dispersion
0.0361g of sodium copper chlorophyllin powder is weighed and added into 100mL of alkaline electrolytic water solution (100 mL of 0.85% NaCl solution is used for the control group) to be mixed evenly by shaking, thus obtaining 500 mu M sodium copper chlorophyllin stock solution for standby.
Determination of ROS with singlet oxygen:
after the mixed species biofilm was incubated for a specified period of time, planktonic bacteria were removed and washed 3 times with PBS. 1mL of copper sodium chlorophyllin solutions (0, 20, 50, 100, 150, 200. Mu.M) of different concentrations were added to the well plate, respectively, wherein 0. Mu.M was replaced with an equal volume of 0.85% NaCl solution or electrolytic aqueous solution. Then 10. Mu.L of ROS reactive oxygen species fluorescent probe (monitoring ROS production) or 100. Mu.L of AMDA reagent (monitoring singlet oxygen production) was added, and the mixture was left to stand and incubate for 10min and then irradiated with light for various times (0, 5, 10, 15, 20, 30 min). Absorbance at wavelengths of 504nm and 399nm was measured with a multifunctional microplate reader, respectively.
Effect of different concentrations of photosensitizer and illumination time on biofilm removal effects of mixed species
After mixed species biofilms were incubated in 24-well polystyrene plates (plastic) for 18h, planktonic bacteria were removed and washed with PBS solution.
(1) Four groups of experiments are set up to explore the cleaning effect of photosensitizers with different concentrations on biological membranes of mixed species in combination with alkaline electrolyzed water. First group (negative control group): 1mL of 0.85% NaCl solution was added; second group (control group): 1mL of sodium copper chlorophyllin with the concentration of 200 mu M and prepared by dissolving 0.85% NaCl solution is added; third group (control group): adding 1mL of alkaline electrolyzed water; fourth group (experimental group): 1mL of sodium copper chlorophyllin prepared by dissolving in alkaline electrolytic aqueous solution was added at various concentrations (20, 50, 100, 150, 200. Mu.M). And the second, third and fourth groups are subjected to static incubation for 10min and then are subjected to illumination for 20min.
(2) Four experiments were also set up to explore the effect of different illumination times in combination with alkaline electrolyzed water on the removal of biofilm from mixed species. First group (negative control group): 1mL of 0.85% NaCl solution was added; second group (control group): 1mL of sodium copper chlorophyllin with the concentration of 150 mu M and prepared by dissolving 0.85% NaCl solution is added; third group (control group): adding 1mL of alkaline electrolyzed water; fourth group (experimental group): 1mL of sodium copper chlorophyllin prepared by dissolution in an alkaline electrolytic aqueous solution at a concentration of 150. Mu.M was added. The second group and the third group are subjected to static incubation for 10min and then are subjected to illumination for 30min, and the fourth group is subjected to static incubation for 10min and then are subjected to illumination for 5min, 10min, 15 min, 20min and 30min respectively.
The remaining biofilm of all the treatment groups was quantified by crystal violet method, and the specific procedures were as follows: after the completion of the treatment, the cells were washed with PBS to remove plankton, and dried in an oven at 55℃to add 1mL of 0.1% (v/v) crystal violet solution to each well, and allowed to stand at room temperature for 30 minutes. The solution was washed 3 times with PBS to remove excessive crystal violet, and 1mL of 95% ethanol was added to each well and allowed to stand at room temperature for 30min, followed by measuring its absorbance at 600nm with a multifunctional microplate reader.
MTT method and plate counting method for characterizing removal effect of biological envelope of mixed species
Similarly, four sets of experiments were set up. A first group: adding 0.85% NaCl solution for treatment; second group: adding sodium copper chlorophyllin with concentration of 150 μm and prepared by 0.85% NaCl solution for dissolving; third group: adding alkaline electrolyzed water for treatment; fourth group: sodium copper chlorophyllin prepared by dissolving in alkaline electrolytic water solution is added for treatment, wherein the concentration of the sodium copper chlorophyllin is 150 mu M. All groups were incubated for 20min with light, plankton removed and washed with PBS. 1mL of fresh TSB culture medium and 100 mu L of MTT reagent are added into each well, and the mixture is placed in a 25 ℃ incubator for standing and incubation for more than 2 hours. After washing planktonic bacteria with PBS solution, 1mL DMSO solution was added to each well, and the mixture was allowed to stand at room temperature for 30 minutes, and then absorbance at 570nm was measured.
The number of viable bacteria in the mixed species biofilm was detected by plate counting. The treatment of the biofilm was completely identical to that in the MTT method. After the treatment, 1mL of 0.85% NaCl solution was added to each well, and the remaining biofilm cells at the bottom of the well plate were suspended by repeated blowing with a gun head, transferred to a centrifuge tube, and then subjected to gradient dilution and respectively spread on a TCBS plate (12 h/37 ℃) and an iron agar plate (24 h/25 ℃). After incubation for a specified time, counts were taken.
The crystal violet method and the MTT method characterize the cleaning effect on the biological film of the mixed species formed on the stainless steel and the fish scales
The sterilized stainless steel discs and round fish scales are placed at the bottom of a 24-hole plate in advance, 10 mu L of mixed bacterial liquid and 990 mu L of sterile fresh TSB culture medium are added, and then the hole plate is placed in a 25 ℃ incubator for static culture for 18 hours, so that a mixed species biofilm is formed on the surface of the hole plate. And then, the biological film is completely consistent with the MTT method and the plate counting method in the characterization of the removal effect on the mixed biological film, and the biological film is divided into four groups, and the removal effect of the biological film is verified by a crystal violet method and an MTT method respectively after the treatment is finished.
Data analysis
Each set of samples was set up in 3 replicates and averaged after 3 independent replicates. Data processing is performed by Microsoft Excel 2010; single-factor analysis of variance (P < 0.05) was performed by SPSS 21.0; mapping analysis was performed by Graphpad Prism 8.4.3 and Origin 8.0.
A, b, c, d, e, f in fig. 3-5 is automatically generated by the SPSS software in analyzing the data to express significant differences.
Example 1
Effects of PDT in combination with alkaline electrolyzed water on ROS and singlet oxygen
The effect of PDT in combination with alkaline electrolyzed water on ROS and singlet oxygen production levels As shown in FIG. 2, when no photosensitizer treatment was added (0.85% NaCl solution or alkaline electrolyzed water was added), faint ROS levels could be detected in the system, since microorganisms would produce endogenous ROS during normal metabolism (FIGS. 2-A, 2-C). The illumination time was maintained at 20min, and when the photosensitizer concentration was 20. Mu.M, the PDT alone treatment detected a ROS level of 0.101 and the PDT combined with alkaline electrolyzed water treatment detected a ROS level of 0.202. As the photosensitizer concentration increases, ROS in the PDT alone treatment group increases slowly, with ROS levels of only 0.288 when the photosensitizer concentration reaches 200 μm. However, as the photosensitizer concentration increased in the PDT combined alkaline electrolyzed water treatment group, ROS levels increased significantly, reaching 0.646 when the photosensitizer concentration was 200 μm. The growth rate is significantly higher than in the PDT alone treatment group. The effect of light time on ROS levels also has a similar trend. In brief, when no light is applied and the concentration of photosensitizer is maintained at 150. Mu.M, there is no significant difference in ROS levels between the PDT alone treatment group and the PDT in combination with alkaline electrolyzed water treatment group. As the illumination time is prolonged, both ROS levels gradually increase. When the illumination time is 30min, the ROS level of the PDT independent treatment group is only 0.269, however, the ROS level of the PDT combined alkaline electrolyzed water group is obviously higher than that of the PDT independent treatment group, and the ROS level is 0.552 which is more than 2 times that of the PDT independent treatment group.
The ama reagent has a maximum absorbance at 399nm, and singlet oxygen can react with it to deplete the ama. The level of singlet oxygen production is therefore proportional to the decrease in absorbance of the AMDA at 399 nm. As shown in FIGS. 2-B and 2-D, the absorbance of AMDA at 399nm was 1.262 without the addition of a photosensitizer or without illumination. The light irradiation time of both groups was kept at 20min, and when the concentration of sodium copper chlorophyllin was 20. Mu.M, the absorbance value of the PDT-alone treatment group was reduced to 1.176, and the absorbance value of the PDT-combined alkaline electrolyzed water group was reduced to 1.058. As the photosensitizer concentration increased, the absorbance at 399nm was continuously decreased for both groups. The rate of decrease of PDT in combination with alkaline electrolyzed water is significantly higher than in the PDT alone treatment group. When the chlorophyll concentration was increased to 200. Mu.M, the absorbance of the PDT-alone treatment group was decreased to 0.750, and the PDT-combined alkaline electrolyzed water group was decreased to 0.183. The trend of the illumination time on the PDT alone treatment in combination with alkaline electrolyzed water treatment is similar. The above results indicate that after PDT combined with alkaline electrolyzed water, the ROS and singlet oxygen production levels are greatly increased compared to PDT alone, indicating the coupled interactions between PDT and electrolyzed water, which may increase their sterilization efficiency.
Example 2
Effect of PDT in combination with alkaline electrolyzed water on removal of biofilm from mixed species under different conditions
To investigate whether PDT in combination with alkaline electrolyzed water would enhance the removal of mixed species biofilms, we characterized their removal in 24-well plates (plastics) by crystal violet. As shown in FIG. 3-A, the total biofilm biomass of the negative control group was 1.963 (OD 600nm ) When PDT was treated alone (photosensitizer concentration 200. Mu.M, illumination time 20 min), total biomass was reduced by 19.8%; when alkaline electrolyzed water was treated alone, the total biomass was reduced by 42.2%. However, when PDT is combined with alkaline electrolyzed water treatment, the photosensitizer concentration is only 20. Mu.M, and the total biomass is reduced by 46.6%. Further improves the concentration of the photosensitizer and obviously reduces the total biomass of the biofilm. When the photosensitizer concentration was increased to 150 μm, the total biomass had been reduced by 72.6% compared to the control group. The effect of light exposure time on PDT combined with alkaline electrolyzed water to clear biofilm also has a similar trend. Similarly, when the photosensitizer concentration was fixed at 150. Mu.M and the illumination time was 30min, PDT alone reduced the total biofilm biomass by 20.6% and alkaline electrolyzed water alone by 43.2%. However, PDT combined with alkaline electrolyzed water illumination was only 5min, with a total biomass reduction of more than half. After prolonged illumination time, the total biomass is further reduced. When the illumination time reaches 20min, the total biomass is reduced by more than 70 percent. The experimental results above strongly demonstrate that PDT is combined with alkaline electrolysisThe removal efficiency of the mixed species biofilm after water is significantly higher than PDT alone or alkaline electrolyzed water alone. Furthermore, it has been found that the removal efficiency of PDT in combination with alkaline electrolyzed water for biofilm is greater than the sum of the removal efficiency of PDT alone and alkaline electrolyzed water alone. This further verifies that there is a coupling interaction between PDT and alkaline electrolyzed water.
We used the MTT method and plate count method to detect cell viability and total colony count before and after treatment, respectively. As a result, as shown in FIG. 4, in the negative control group, the initial cell viability of the mixed species biofilm was 0.844 (OD 570nm ). When alkaline electrolyzed water was treated alone, the cell viability was reduced to 0.326. When the photosensitizer concentration was 150. Mu.M and the illumination time was 20min, the cell viability was reduced to 0.557. However, when PDT was combined with alkaline electrolyzed water, cell viability was reduced to 0.124 at equivalent conditions, 85.3% compared to the control. Similarly, the plate count results showed a total colony count in the initial mixed species biofilm of 8.28Log CFU/mL (negative control). After alkaline electrolyzed water and PDT were treated separately, the total colony count remained 6.12 and 7.36Log CFU/mL in sequence. When PDT is combined with alkaline electrolyzed water, the total colony number is reduced to 4.01Log CFU/mL, and the sterilization efficiency is as high as more than 99.99%. The results of cell viability and colony count further demonstrate that the experimental results of the above crystal violet method are supported, while also demonstrating the high efficiency of PDT in combination with alkaline electrolyzed water.
Example 3
Cleaning effect of PDT combined with alkaline electrolyzed water on biological membranes of mixed species on different contact surfaces
Stainless steel and fish scales are common adhesion carriers in the actual life of the biofilm. We have therefore further explored the cleaning effect of PDT in combination with alkaline electrolyzed water on biofilm of mixed species adhering to the surfaces of stainless steel and fish scales. As shown in FIGS. 5-A and 5-B, the total biomass of the stainless steel and the scale surface biofilm in the negative control group was 1.79 and 3.94 (OD 600nm ). When PDT was treated alone (photosensitizer concentration: 150. Mu.M; light time: 20 min), there was no significant difference between total biomass and control. When treated with alkaline electrolyzed water alone,the total biomass of the biofilm adhered to the stainless steel is reduced by 37 percent, and the total biomass of the biofilm adhered to the fish scales is still not obviously reduced. However, when PDT was combined with alkaline electrolyzed water treatment, the total biofilm biomass adhering to stainless steel and fish scales was reduced by 52.1% and 13.6%, respectively. We have found that the removal efficiency of PDT in combination with alkaline electrolyzed water is not high by crystal violet method, because crystal violet not only can bind to biofilm but also can stain fish scales, resulting in inaccurate results. Therefore, we further explored the variation in biofilm colony count by plate counting. As shown in FIG. 5-C and FIG. 5-D, the total number of biofilm colonies on the surfaces of the stainless steel and the fish scales in the negative control group was 8.11 and 8.51Log CFU/mL, respectively. When treated with PDT alone, neither was significantly degraded; when treated with alkaline electrolyzed water alone, the total number of biofilm colonies on the surfaces of stainless steel and fish scales was 6.83 and 8.29Log CFU/mL, respectively. However, when PDT is combined with alkaline electrolysis water treatment, the total number of biofilm colonies on the two contact surfaces is significantly reduced, and the sterilization rates reach 99.93% and 99.22%, respectively. These results demonstrate that PDT combined with alkaline electrolyzed water also has good cleaning effect on the biofilm on the surfaces of stainless steel and fish scales, which further reflects the practical application value of PDT combined with alkaline electrolyzed water in the food industry. Therefore, the technology is expected to become a new means for controlling the pollution of harmful microorganisms in the food industry in the future.
Proved by verification, the photodynamic technology is combined with the alkaline electrolyzed water to improve the sterilization efficiency, creatively combines the photodynamic technology with the electrolyzed water technology, and opens up a brand-new channel for improving the PDT sterilization efficiency; the technology can be applied to the food industry by matching alkaline electrolyzed water with a food-grade photosensitizer, so that the risk brought by pollution of harmful microorganisms is reduced; the method has the advantages of simple operation, good applicability, low cost, good safety, green and pollution-free performance, and capability of remarkably improving the sterilization efficiency (on vibrio parahaemolyticus and Shewanella putrefying) and great application prospect.
While particular embodiments of the present invention have been described above, it will be understood by those skilled in the art that these are by way of example only and that various changes or modifications may be made to these embodiments without departing from the principles and spirit of the invention.
Claims (4)
1. A method for improving sterilization efficiency by combining photodynamic technology with alkaline electrolyzed water is characterized by comprising the following steps:
(1) Mixing a photosensitizer dispersion with a sample to be treated, wherein the photosensitizer dispersion comprises a photosensitizer and alkaline electrolyzed water;
(2) Incubating the mixed sample, wherein the incubation is performed under a dark condition for 10-60 min;
(3) Irradiating the incubated sample with a blue light source with wavelength of 455-460 nm and light power density of 2-50 mW/cm 2 The method comprises the steps of carrying out a first treatment on the surface of the The irradiation duration of the light source is 20-120 min;
wherein the photosensitizer is sodium copper chlorophyllin;
the preparation method of the alkaline electrolyzed water comprises the following steps:
preparing alkaline electrolyzed water by using an electrolyzed water preparation instrument, wherein a water inlet is common tap water, a water outlet is target electrolyzed water, and parameters of the electrolyzed water preparation instrument are as follows: ph=9 to 10, oxidation-reduction potential= -300 to-400 mV;
the concentration of the photosensitizer in a sample mixed system to be treated is 80-1000 mu M;
the improvement of the sterilization efficiency means improvement of the sterilization efficiency against vibrio parahaemolyticus and Shewanella putrefying.
2. A photosensitizer dispersion for use in a method for improving sterilization efficiency by combining photodynamic technology with alkaline electrolyzed water as described in claim 1, characterized by comprising a photosensitizer and alkaline electrolyzed water,
wherein the photosensitizer is sodium copper chlorophyllin;
the preparation method of the alkaline electric alkali liquid comprises the following steps:
preparing alkaline electrolyzed water by using an electrolyzed water preparation instrument, wherein a water inlet is common tap water, a water outlet is target electrolyzed water, and parameters of the electrolyzed water preparation instrument are as follows: ph=9 to 10, oxidation-reduction potential= -300 to-400 mV.
3. Use of a photosensitizer dispersion according to claim 2 for sterilization of vibrio parahaemolyticus and shiwanella putrefying bacteria.
4. Use of a photosensitizer dispersion according to claim 2 for removing biofilm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110382052.XA CN113068791B (en) | 2021-04-09 | 2021-04-09 | Method for improving sterilization efficiency by combining photodynamic technology with alkaline electrolyzed water |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110382052.XA CN113068791B (en) | 2021-04-09 | 2021-04-09 | Method for improving sterilization efficiency by combining photodynamic technology with alkaline electrolyzed water |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113068791A CN113068791A (en) | 2021-07-06 |
| CN113068791B true CN113068791B (en) | 2024-04-16 |
Family
ID=76615750
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110382052.XA Active CN113068791B (en) | 2021-04-09 | 2021-04-09 | Method for improving sterilization efficiency by combining photodynamic technology with alkaline electrolyzed water |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113068791B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114403209A (en) * | 2021-12-27 | 2022-04-29 | 中国农业大学 | Method for processing plant source food raw material |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108450478A (en) * | 2018-04-04 | 2018-08-28 | 武汉珞格普润生态技术有限公司 | A method of preventing grey mould fruit rot of strawberry using acidic oxidation electrolysis water |
| CN109730239A (en) * | 2019-03-20 | 2019-05-10 | 西南科技大学 | A kind of bactericidal liquid and preparation method thereof containing electrolyzed alkaline water |
| CN111067007A (en) * | 2019-12-26 | 2020-04-28 | 上海海洋大学 | Method for killing salmonella through photodynamic |
-
2021
- 2021-04-09 CN CN202110382052.XA patent/CN113068791B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108450478A (en) * | 2018-04-04 | 2018-08-28 | 武汉珞格普润生态技术有限公司 | A method of preventing grey mould fruit rot of strawberry using acidic oxidation electrolysis water |
| CN109730239A (en) * | 2019-03-20 | 2019-05-10 | 西南科技大学 | A kind of bactericidal liquid and preparation method thereof containing electrolyzed alkaline water |
| CN111067007A (en) * | 2019-12-26 | 2020-04-28 | 上海海洋大学 | Method for killing salmonella through photodynamic |
Non-Patent Citations (4)
| Title |
|---|
| Effects of the curcumin-mediated photodynamic inactivation on the quality of cooked oysters with Vibrio parahaemolyticus during storage at different temperature;Bowen Chen等;International Journal of Food Microbiology;第345卷;1-11 * |
| Eradication of planktonic Vibrio parahaemolyticus and its sessile biofilm by curcumin-mediated photodynamic inactivation;Bowen Chen等;Food Control;第113卷;1-10 * |
| 光敏杀菌技术及其在食品中的研究与应用;李天密;屈思佳;高文惠;赵玲钰;韩俊华;;保鲜与加工(06) * |
| 碱性电解水对罗非鱼活体杀菌工艺优化;梁凤雪;陈海强;梁钻好;敖菲菲;冯均元;李建钰;余铭;;农业工程(05) * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113068791A (en) | 2021-07-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Varghese et al. | Antibacterial efficiency of carbon dots against Gram-positive and Gram-negative bacteria: A review | |
| Sattarahmady et al. | Bactericidal laser ablation of carbon dots: An in vitro study on wild-type and antibiotic-resistant Staphylococcus aureus | |
| Nitzan et al. | ALA induced photodynamic effects on gram positive and negative bacteria | |
| Caminos et al. | Photodynamic inactivation of Escherichia coli by novel meso-substituted porphyrins by 4-(3-N, N, N-trimethylammoniumpropoxy) phenyl and 4-(trifluoromethyl) phenyl groups | |
| Wu et al. | Synthesis of high-performance conjugated microporous polymer/TiO2 photocatalytic antibacterial nanocomposites | |
| Wang et al. | Wound therapy via a photo-responsively antibacterial nano-graphene quantum dots conjugate | |
| Pezzoni et al. | Protective role of extracellular catalase (KatA) against UVA radiation in Pseudomonas aeruginosa biofilms | |
| CN113068791B (en) | Method for improving sterilization efficiency by combining photodynamic technology with alkaline electrolyzed water | |
| CN112640923B (en) | Application of turmeric essential oil as ultraviolet sterilization synergist | |
| CN113087863B (en) | Efficient photodynamic sterilization porphyrin covalent organic framework material and preparation method thereof | |
| Wang et al. | Photocatalytic inactivation and destruction of harmful microalgae Karenia mikimotoi under visible-light irradiation: Insights into physiological response and toxicity assessment | |
| Reynoso et al. | Photoactive antimicrobial coating based on a PEDOT-fullerene C 60 polymeric dyad | |
| Pudziuvyte et al. | Alterations of Escherichia coli envelope as a consequence of photosensitization with tetrakis (N-ethylpyridinium-4-yl) porphyrin tetratosylate | |
| Pinto et al. | Investigation of powerful fungicidal activity of tetra-cationic platinum (II) and palladium (II) porphyrins by antimicrobial photodynamic therapy assays | |
| Wang et al. | Metal-free nitrogen-doped carbon nanodots as an artificial nanozyme for enhanced antibacterial activity | |
| Giannakis et al. | Identifying the mediators of intracellular E. coli inactivation under UVA light: The (photo) Fenton process and singlet oxygen | |
| CN114917363B (en) | Nanocomposite and preparation method and application thereof | |
| Street et al. | Eradication of the corrosion-causing bacterial strains Desulfovibrio vulgaris and Desulfovibrio desulfuricans in planktonic and biofilm form using photodisinfection | |
| Zhao et al. | Bacterial microenvironment-responsive dual-channel smart imaging-guided on-demand self-regulated photodynamic/chemodynamic synergistic sterilization and wound healing | |
| Hernández-Zanoletty et al. | Assessment of new immobilized photocatalysts based on Rose Bengal for water and wastewater disinfection | |
| Oriel et al. | Mechanistic aspects of photoinactivation of Candida albicans by exogenous porphyrins | |
| Hong et al. | Mechanisms of Escherichia coli inactivation during solar-driven photothermal disinfection | |
| Tang et al. | Rapid and complete inactivation of pathogenic microorganisms by solar-assisted in-situ H2O2 generation using a polypyrrole-supported copper sulfide system | |
| Liu et al. | Chiral organic nanoparticles based photodynamic antibacterial films for food preservation | |
| CN116270480A (en) | Caffeic acid metal polyphenol coated metal-organic framework nanoparticle and preparation method and application thereof |
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 |