CN105232574B - Acidic electrolytic water and its production and use - Google Patents
Acidic electrolytic water and its production and use Download PDFInfo
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Abstract
The present invention relates to food processing technology field, more particularly to a kind of acidic electrolytic water and its production and use, the chloro electrolyte that wherein it is 0.05~0.1% by mass concentration that acidic electrolytic water, which is, has the acidic electrolytic water that pH value is 2.7~3.0, effective chlorine density is 55~80mg/L, oxidation-reduction potential is 1150~1180mV what anode region electrolysis obtained.Compared with prior art, the acidic electrolytic water of the present invention is in low pH, under the three of high redox potential and high effective chlorine density synergy, to food-borne pathogens biofilm, particularly the bacillus cereus biofilm of better resistance has very strong killing effect, the invention provides the new application that acidic electrolytic water kills food-borne pathogens biofilm, available for the pharmaceutical composition for preparing killing food-borne pathogens biofilm, food additives composition and disinfectant or cleaning agent, for the control of biofilm in food industry, removing provides new approach, and then provide reliable guarantee for the security of food.
Description
Technical field
The present invention relates to food processing technology field, and in particular to a kind of acidic electrolytic water and its production and use.
Background technology
With the development of society, the raising of living standards of the people, food security is increasingly valued by people.But no matter
It is in developing country or developed country, human health all suffers from the threat of food origin disease.According to statistics, the annual whole world is big
The experience of about 1/3rd people's criminal's of having food origin disease.
Foodborne bacterial pathogenses are to cause food spoilage and trigger the principal element of food origin disease, and food security to face
Long standing difficulty.At present, research work both domestic and external is concentrated mainly on to dissociate in food and causes a disease, causes rotten microorganism aspect.However,
Increasing research shows, is food peace the problems such as bacterial biofilm, the reparation of sub- lethal cell and the variation of virus
Complete important recessive harm, and the major reason for causing food origin disease to break out.It is estimated that about 65%-80% bacteriums are sexy
Dye is (Hall-Stoodley L, Costerton J W, the Stoodley P.Bacterial as caused by biofilm
biofilms:from the natural environment to infectious diseases[J].Natural
Reviews Microbiology,2004,(2):95-108).Wherein, the biofilm formed in food industry is most likely to be
The arch-criminal of food origin disease.For example, in food fermentation, some spoilage organisms and pathogen can form biofilm, so as to
Cause corruption and the outburst of disease of final products.Therefore, the food-safe threat of biofilm is not allow in food industry
Ignore.
The biofilm distinctive biological phenomena that is microorganism for reform of nature environment, is being unfavorable for its growth or existing
Under the conditions of, it is inter-adhesive between bacterium, and secrete polymer substrate and form membranoid substance.Any bacterium under given conditions can shape
Into biofilm, common envelope pathogenic bacteria include listeria monocytogenes, Escherichia coli, waxy gemma bar
The biofilms such as bacterium, staphylococcus aureus, enterococcus faecalis, Klebsiella pneumoniae and pseudomonas aeruginosa.Wherein, it is waxy
Bacillus is a kind of common pathogeny bacterium that can cause food poisoning, is widespread in nature, and easily pollution is rich in albumen
The rice of matter and starch, meat products, bean food, dairy products and bakery product etc..The microbial sitotoxismus case is at me
Ranked among the best in state's food poisoning.But so far, it is domestic there is not yet the report studied on bacillus cereus biofilm
Road；Correlative study is also less abroad.
Compared with plankton, biofilm is stronger to the resistance of environmental pressure and harm is bigger, is more difficult to remove.According to report
Road, biofilm are 1/10~1/1000 (Gristina of flcating germ of the same race to the sensitivity of various chemical bactericides
A.G,Hobgood C D,Webb L X,et al.Adhesive colonization of biomaterials and
antibiotic resistance[J].Biomaterials,1987,8(6):423-426).Because biofilm is to disinfectant
Resistance it is stronger, therefore biofilm is such as brewageed to food industry, marine products processing, Dairy Processing, poultry dressing farm and meat add
Work etc. brings serious food-safety problem.It is any stage of food processing, all kinds of from food production to equal may be infected is consumed
Microorganism, particularly spoilage organisms and pathogen are easily adhered to food surface, and biofilm is formed in food processing system.Biology
Envelope once formed, not only can contaminated food products, increase equipment cleaning difficulty, make purging system failure on the spot, it is harmful to turn into other
It the hiding-place of microorganism, can also corrode food processing and storage and transportation equipment, bring serious harm.In addition, biofilm can also promote
Make more bacterium collaboration symbiosis, equipment surfaces externally and internally is turned into the recessive biohazard source for propagating food origin disease, cause to be difficult to
The Food Quality and Safety problem of solution.Therefore, in food processing process, the control and removing of biofilm are to ensureing food peace
It is complete particularly important.
At present, it is still pathogen in food and its process equipment that the main direction of studying in food industry, which is, i.e.,
Pathogen in food is killed using chemical reagent such as disinfectant or bactericide.Generally lacking organic matter (such as fat, carbon water
Compound, protein etc.) under conditions of, the sterilization effect of disinfectant is more preferable.In addition, organism kinds, pH, temperature, water is hard
Degree, chemical inhibitor, decontaminant concentration and time of contact are also (Bremer P J, Monk an important factor for influenceing Disinfection Effect
I,Butler R.Inactivation of Listeria monocytogenes/Flavobacterium spp.biofilms
using chlorine:impact of substrate,pH,time and concentration[J].Letters in
Applied Microbiology,2002,35(4):321-325).Saitou etc. have studied different bactericide (alcohol, Xian Bi
Too, 10% PVP-I, 0.1% benzalkonium chloride and 0.1% alkyl-dimethyl guanidine-acetic acid atarax) to pseudomonas aeruginosa biology quilt
The killing effect of film, find in addition to 0.1% alkyl-dimethyl guanidine-acetic acid atarax, killing rate of remaining 5 kinds of bactericide to flcating germ
Almost 100%；And it is constant to envelope state bacterium, the killing rate of only 0.1% benzalkonium chloride, the bactericidal effect of remaining bactericide is equal
Decline significantly (Saitou K, Furuhata K, Kawakami Y, et al.Biofilm formation abilities and
disinfectant-resistance of Pseudomonas aeruginosa isolated from cockroaches
captured in hospitals[J].Biocontrol Science,2009,14(2):65-68).Therefore, bactericide can
The common pathogen in food is killed but to be difficult to kill envelope state bacterium and its biofilm.Furthermore food industry is to bactericide
Using there is a strict requirements, bactericide must safely, effectively, it is easy to use, be finished after can be removed from carrier surface, it is non-toxic residual
Stay.But traditional chemical bactericide only high concentration, it is heavy dose of when just have more significant killing action to biofilm, it is and highly concentrated
Degree, the use of heavy dose of chemical bactericide constitute a serious threat to human health.In addition, some bacteriums may be dashed forward by gene
The modes such as change produce certain resistance and physiology plasticity to bactericide.Therefore, in food processing process, the prevention and control of biofilm
Make very urgent.
Electrolysis water is function caused by dilute salting liquid is electrolysed in an electrolytic cell for being separated positive and negative electrode as film
Water.After electrolysis, the physicochemical property such as its pH, oxidation-reduction potential (ORP), effective chlorine density (ACC) occurs notable dilute salting liquid
Change.Wherein, caused function water is acidic electrolytic water in positive pole pond, has strong oxidizing property and antibacterial activity；In negative pole pond
In caused function water be electrolyzed alkaline water, there is strong reducing property, its antibacterial activity is relatively low.Acute and sub-acute toxicity test is
Show that acidic electrolytic water is harmless, color, transparent, without obvious irritation smell safety-type bactericide, for killing resistance to first
Staphylococcus aureus (Huang Y R, Hung Y C, Hsu S Y, the et al.Application of of the woods of oxygen four
electrolyzed water in the food industry[J].Food Control,2008,19(4):329-345)。
Compared with traditional chemical bactericide, acidic electrolytic water has that bactericidal effect is good, cost is low, advantages of environment protection, extensive
Applied to fields such as food, agricultural, medical treatment, livestock products, aquatic products.
In the prior art, the research for acidic electrolytic water is primarily directed to food-borne pathogens such as Escherichia coli, golden yellow
The equal killing effect such as color staphylococcus, Listeria Monocytogenes, Bacterium enteritidis, campylobacter jejuni is ground
Study carefully.The wide spectrum of acidic electrolytic water, efficient bactericidal property are confirmed by domestic and international research, but the bactericidal property of acidic electrolytic water
Easily disturbed by organic matter, research shows that organic interfering substance can reduce ACC and ORP rapidly, so as to weaken acidic electrolytic water significantly
Bactericidal property (Ayebah B, Hung Y C, Kim C, et al.Efficacy of electrolyzed water in the
inactivation of planktonic and biofilm Listeria monocytogenes in the presence
of organic matter[J].Journal of Food Protection,2006,69(9):2143-2150).It is for example, sour
Active chlorine compound in property electrolysis water is easily far below Cl with albumen qualitative response formed with machine chloramines, its antibacterial activity2。
However, relevant acidic electrolytic water causes for killing food-borne pathogens biofilm especially for envelope at present
The research of germ (bacillus cereus) biofilm and its killing mechanism still belongs to blank.Therefore, tool can be killed by needing research badly
The technology of resistant envelope pathogenic bacteria biofilm strong, harm is big, so as to be controlled, clearly for biofilm in food industry
Remove and ensure food safety and new approach is provided.
The content of the invention
An object of the present invention is in view of the shortcomings of the prior art, there is provided one kind can significantly kill waxy gemma bar
The food-borne pathogens such as bacterium biofilm and cost is low, security is good, the acidic electrolytic water that is readily produced.
The second object of the present invention is in view of the shortcomings of the prior art, there is provided one kind can significantly kill waxy gemma bar
The food-borne pathogens such as bacterium biofilm and cost is low, security is good, the preparation method of acidic electrolytic water that is readily produced.
The third object of the present invention is in view of the shortcomings of the prior art, there is provided a kind of acidic electrolytic water is killing waxy bud
The new application of the food-borne pathogens biofilm such as spore bacillus, available for the medicine for preparing killing food-borne pathogens biofilm
Composition, food additives composition and disinfectant or cleaning agent.
To achieve these goals, the present invention adopts the following technical scheme that：
A kind of acidic electrolytic water is provided, the acidic electrolytic water is electrolysed by the chloro that mass concentration is 0.05~0.1%
What matter was electrolysed acquisition in anode region is 2.7~3.0 with pH value, effective chlorine density is 55~80mg/L, oxidation-reduction potential is
1150~1180mV acidic electrolytic water.
Preferably, the pH value of the acidic electrolytic water is 2.7~2.8.
Preferably, the effective chlorine density of the acidic electrolytic water is 70~80mg/L.
Preferably, the oxidation-reduction potential of the acidic electrolytic water is 1170~1180mV.
Preferably, the chloro electrolyte is the sodium chloride that mass concentration is 0.05~0.1%.
The present invention also provides the preparation method of above-mentioned acidic electrolytic water, comprises the following steps：
(1) it is electrolysed：
In a cell, it is 0.05~0.1% chloro electrolyte solution as electrolyte using mass concentration, electrolysis 10~
15min, obtained in anode region pH value be 2.7~3.0, effective chlorine density is 55~80mg/L, oxidation-reduction potential be 1150~
1180mV acidic electrolytic water；
In electrolytic process, per 5min using the effective chlorine density of indirect iodometric processes measure anode region electrolysis water, pH meter measure
PH value, oxidation-reduction potential test pen measure oxidation-reduction potential；
(2) lucifuge storage is sealed：
Acidic electrolytic water prepared by step (1), under the conditions of lucifuge, is positioned over sealed storage in 35 DEG C of incubators, stores up
The time is deposited no more than 9 days.
It is used for the new application for killing food-borne pathogens biofilm present invention also offers above-mentioned acidic electrolytic water.
The acidic electrolytic water kills the pharmaceutical composition or food additives of food-borne pathogens biofilm preparing
Application in composition.
Application of the acidic electrolytic water in the disinfectant or cleaning agent of killing food-borne pathogens biofilm is prepared.
The beneficial effects of the invention are as follows：
The acidic electrolytic water of the present invention, it is to be electrolysed by the chloro electrolyte that mass concentration is 0.05~0.1% in anode region
Obtain have pH value be 2.7~3.0, effective chlorine density is 55~80mg/L, oxidation-reduction potential is 1150~1180mV's
Acidic electrolytic water.Compared with prior art, the acidic electrolytic water that prepared by the present invention has advantages below：
1) acidic electrolytic water of the invention has higher effective chlorine density (55~80mg/L), either high redox electricity simultaneously
Position (1150~1180mV) and the faintly acid that pH value is 2.7~3.0, in low pH, high redox potential and effective chlorine density
Three synergy under, so as to bacillus cereus this kind of better resistance food-borne pathogens biofilm have very
Strong killing effect；
2) preparation method of the invention is simple, cost is relatively low, be readily produced and security is good, lucifuge, 35 DEG C and
Under confined conditions, there is preferable storage stability, the influence to pH, oxidation-reduction potential and higher effective chlorine density value is very
It is small, the effect of can storing 0~9 day, and can still keep stronger killing food-borne pathogens biofilm；
3) the invention provides the new application that acidic electrolytic water kills food-borne pathogens biofilm, due to acidic electrolysis
The security of water is good, available for pharmaceutical composition, the food additives composition for preparing killing food-borne pathogens biofilm
And disinfectant or cleaning agent, for the control of biofilm in food industry, the new approach that provides is removed, and then be food
Security provides reliable ensure.
Embodiment
The invention will be further described with the following Examples.
Following examples use the bacillus cereus (culture presevation of Guangdong Microbes Inst DSMZ
Number：CMCC63303), tested by bacteria suspension quantitative sterilization.
1 experiment material and instrument and equipment
Hydrochloric acid, sulfuric acid, KI, sodium thiosulfate etc. are to analyze pure or chemical reagent；
Nertralizer forms：0.5% sodium thiosulfate PBS (0.03M, pH 7.2)；
CE-7001 type acidic oxidized electric potential water generators are provided by Guangzhou Saiai Environment Protection Technology Development Co., Ltd.
2 experimental methods
The preparation of 2.1 bacteria suspensions
After bacillus cereus is activated, it is inoculated in sterile broth bouillon, 37 DEG C of culture 12h to viable count are
108CFU/mL, it is standby.
The preparation of 2.2 biofilms
(1) cleaning of carrier-pellet：Carrier-pellet is rinsed with Natural Water, puts ultrasonic 15min (supersonic frequencies in acetone
40kHz, ultrasonic power 250W), remove surface and oil contaminant；Then, it is placed in 75% ethanol and soaks 30min, then with washing
Agent is cleaned, finally clean with distilled water flushing, is sterilized after drying standby.
(2) culture of biofilm：Carrier-pellet and 10mL broth bouillons are added into test tube, after sterilizing, accesses 1%
Above-mentioned bacteria suspension (108CFU/mL), cultivated at 37 DEG C.
The measure of 2.3 biofilm viable counts
Using ultrasonic the method for plate culture count：After biofilm culture certain time, the liquid medium in test tube is outwelled,
Then rinsed 3 times with sterile PBS (0.05M, pH 7.2), remove the bacterium of flcating germ and adhesion loosely, then added with Sterile pipette
Enter the sterile PBS of 10mL (0.05M, pH 7.2), and ultrasonic 15min (250W), plate count then is carried out to bacteria suspension, put down
Row experiment 3 times, statistical analysis uses Microsoft Office Excel softwares.
Biofilm bacterium number is calculated by following equation：
The measure of 2.4 acidic electrolytic water physicochemical properties
In electrolytic process, pH, ACC and ORP of measure anode region electrolysis water per 5min.
2.4.1 the measure of effective chlorine density (ACC)
Indirect iodometric processes：Prepare 2M sulfuric acid, 100g/L KIs, 5g/L starch solutions, 0.005M sodium thiosulfate standards
Solution.
Add 2M sulfuric acid, 100g/L liquor kalii iodide and each 10mL of electrolysis water into 100mL iodine flasks.Now, solution goes out
Existing yellow.Addend drips distilled water in bottle cap edge after covering bottle cap and shaking mixing, puts dark place 5min.Bottle cap is opened, allows Gai Yuan
Distilled water is flowed into bottle.Free-iodine is titrated with sodium thiosulfate standard solution, is shaken up in drop.Treat that solution shoals in yellow, add
The starch solution 10 for entering 5g/L drips, and solution becomes blueness immediately, continues to be titrated to blue disappearance with sodium thiosulfate standard solution, remembers
The sodium thiosulfate standard solution total amount employed, and titration results are corrected with blank test.
Because 1mL 1M sodium thiosulfate standard solutions are equivalent to 0.03545g effective chlorine, available chlorine content is calculated as follows.
In formula：X- available chlorine contents, g/L
C- Concentration of Sodium Thiohsulfate Standard Solution, M
Vst- titration spends the volume of sodium thiosulfate standard solution, mL
The quality of contained disinfectant active compound, g in m- iodine flasks
The volume of the stoste of liquid containing liquid disinfection, mL in V- iodine flasks
2.4.2 the measure of pH and oxidation-reduction potential (ORP)
PH meter and oxidation-reduction potential test pen measure is respectively adopted in pH and oxidation-reduction potential.
2.5 acidic electrolytic waters kill the processing method of bacillus cereus biofilm
The stainless steel substrates for being loaded with culture 24h biofilms are rinsed 3 times with sterile PBS (0.05M, pH 7.2), removed floating
The bacterium of bacterium and adhesion loosely is swum, the acidic electrolytic water prepared by 10mL is added and stands 15min at room temperature, rapidly by stainless steel substrates
It is transferred in 10mL nertralizers and acts on 10min, then rinse stainless steel substrates 3 times with sterile PBS (0.05M, pH 7.2), adds 10mL
Sterile PBS (0.05M, pH 7.2), 15min (250W) is ultrasonically treated, plate count, parallel test 3 are carried out to bacteria suspension
It is secondary.The detection of ultrasonic the method for plate culture count is limited to 0.54log CFU/cm2(i.e. 10CFU/ stainless steel substrates), represent to test with ND
As a result it is less than test limit.Data are carried out using Microsoft Office Excel softwares and SPSS Statistics softwares
Statistical analysis.
Embodiment 1.
Using CE-7001 type acidic oxidized electric potential water generators, electricity is used as by 0.1% sodium chloride solution of mass concentration
Liquid is solved, 10min is electrolysed in electrolytic cell, pH 2.73 is obtained in anode region, the acidity electricity that ACC 79.6mg/L, ORP are 1179mV
Xie Shui, freshly prepd acidic electrolytic water is then taken, be divided in the vial of 1000mL browns, stopper bottle stopper, be put into 35 DEG C
In incubator, storage time is no more than 9 days.
Embodiment 2.
Using CE-7001 type acidic oxidized electric potential water generators, electricity is used as by 0.1% sodium chloride solution of mass concentration
Liquid is solved, 10min is electrolysed in electrolytic cell, pH 2.75 is obtained in anode region, the acidity electricity that ACC 73.7mg/L, ORP are 1174mV
Xie Shui, freshly prepd acidic electrolytic water is then taken, be divided in the vial of 1000mL browns, stopper bottle stopper, be put into 35 DEG C
In incubator, storage time is no more than 9 days.
Embodiment 3.
Using CE-7001 type acidic oxidized electric potential water generators, electricity is used as by 0.1% sodium chloride solution of mass concentration
Liquid is solved, 10min is electrolysed in electrolytic cell, pH 2.78 is obtained in anode region, the acidity electricity that ACC 71.3mg/L, ORP are 1170mV
Xie Shui, freshly prepd acidic electrolytic water is then taken, be divided in the vial of 1000mL browns, stopper bottle stopper, be put into 35 DEG C
In incubator, storage time is no more than 9 days.
Test example 1：
Acidic electrolytic water (pH 2.73, ACC 79.6mg/L, ORP 1179mV) prepared by embodiment 1 uses deionization
Water is diluted to 80%, 60%, the 40% of initial concentration respectively, obtains different ACC acidic electrolytic water, after taking 100mL to dilute
Acidic electrolytic water, add 40,80,100 μ L 1M hydrochloric acid solution thereto respectively, make it consistent with the pH of electrolysis water stoste.
During this, the ORP of acidic electrolytic water is basically unchanged, finally obtain ACC be respectively 51.3,33.6,18.6mg/L acidity electricity
Solve water a, b, c.Be respectively 51.3 using ACC, 33.6,18.6mg/L acidic electrolytic water a, b, c kill bacillus cereus life
Thing envelope, for step with 2.5, electrolysis water action time is 15min.
Test example 2：
The acidic electrolytic water (pH 2.73, ACC 79.6mg/L, ORP 1179mV) prepared to 100mL embodiments 1 is respectively
Add 350,375,400 μ L 1M sodium hydroxide solution, obtain pH be respectively 3.67,4.76,5.82 acidic electrolytic water d, e,
F, in the process, ACC and the ORP change of acidic electrolytic water are smaller.It is respectively 3.67,4.76,5.82,6.66 acid using pH
Property electrolysis water d, e, f kill bacillus cereus biofilm, for step with 2.5, electrolysis water action time is 15min.
Test example 3：
(pH 2.73, ACC 79.6mg/L, ORP are acidic electrolytic water prepared by 3% bromine water and p embodiments 1
1179mV) respectively with 1：9 and 9：1 ratio allotment, obtains different ORP acidic electrolytic water.100mL is taken with 1：The electricity of 9 allotments
Xie Shui, 80 μ L sodium hypochlorite are added thereto, obtain the acidic electrolytic water g that ORP is 976mv.100mL is taken with 9：The electricity of 1 allotment
Xie Shui, 1mL sodium hypochlorite and 3mL 1M sodium hydroxide are added thereto, obtain the acidic electrolytic water h that ORP is 881mv.Utilize
Acidic electrolytic water g, h that ORP is respectively 976,881mv kill bacillus cereus biofilm, and step with 2.5, make by electrolysis water
It is 15min with the time.
Result of the test and analysis：
(1) bin stability of acidic electrolytic water kills the influence of bacillus cereus biofilm to acidic electrolytic water
Although acidic electrolytic water has the advantages that cost is low, environment-friendly, safe, its shortcoming is that stability is bad.
Based on the change of acidic electrolytic water main physical and chemical in storage process, freshly prepd acidic electrolytic water is taken, point
Mounted in 1000mL is transparent and the vial of brown in, stopper bottle stopper, be respectively put into 20,35,50 DEG C of incubator and store 28d,
Timing power taking solution water sample, determines its pH, ACC and ORP.
The stainless steel substrates for being loaded with culture 24h biofilms are rinsed 3 times with sterile PBS (0.05M, pH 7.2), removed floating
The bacterium of bacterium and adhesion loosely is swum, and the acidic electrolytic water stored is added under 10mL different conditions, and standing acts on 15min at room temperature, fast
Stainless steel substrates are transferred in 10mL nertralizers by speed acts on 10min, then rinses stainless steel substrates with sterile PBS (0.05M, pH 7.2)
3 times, the sterile PBS of 10mL (0.05M, pH 7.2) are added, are ultrasonically treated 15min (250W), flat-plate bacterial colony meter is carried out to bacteria suspension
Number, parallel test 3 times.
As a result show, the ACC and ORP of acidic electrolytic water reduce with the extension of period of storage, and pH is with period of storage
Extend and increase；And change in lucifuge, 35 DEG C acidic electrolytic water pH, ORP, the ACC of storage under confined conditions no more than 9 days
Very little.With the extension of period of storage, due to Cl2Volatilization and HOCl decomposition cause ACC to significantly reduce, can largely reducing
The activity of the bacillus cereus biofilm that goes out of acidic electrolytic water.
(2) killing effect of the acidic electrolytic water to bacillus cereus biofilm
The stainless steel substrates for being loaded with culture 24h biofilms are rinsed 3 times with sterile PBS (0.05M, pH 7.2), removed floating
Bacterium and adhesion bacterium loosely are swum, 10mL embodiments 1 and the acidic electrolytic water prepared by test example is separately added into, distinguishes at room temperature
Stainless steel substrates, are transferred in 10mL nertralizers act on 10min rapidly, then with sterile by standing effect 1,3,5,10,15,20min
PBS (0.05M, pH 7.2) rinsings stainless steel substrates 3 times, add the sterile PBS of 10mL (0.05M, pH 7.2), are ultrasonically treated 15min
(250W), plate count, parallel test 3 times are carried out to bacteria suspension.The detection of ultrasonic the method for plate culture count is limited to
0.54log CFU/cm2(i.e. 10CFU/ stainless steel substrates), represent that experimental result is less than test limit with ND.Use Microsoft
Office Excel softwares and SPSS Statistics softwares carry out statistical analysis to data.The acidity electricity of embodiment 1 to 3
Xie Shui is as shown in table 1 to the killing effect of bacillus cereus biofilm.
Killing effect of the acidic electrolytic water of the embodiment 1 to 3 of table 1 to bacillus cereus biofilm
As shown in Table 1, the acidic electrolytic water that prepared by the present invention is to food-borne pathogens biofilm, particularly waxy gemma
The biofilm of the envelope bacterium of this kind of better resistance of bacillus has very strong killing effect.
(3) research of killing mechanism of the acidic electrolytic water to bacillus cereus biofilm
1st, the killing effect of acidic electrolytic water and chemical modification water to bacillus cereus biofilm is contrasted
The stainless steel substrates for being loaded with culture 24h biofilms are rinsed 3 times with sterile PBS (0.05M, pH 7.2), removed floating
The bacterium of bacterium and adhesion loosely is swum, is separately added into 10mL deionized waters (blank), the acidic electrolytic water of embodiment 1,0.05% chlorine
+ 0.05% sour sodium (ACC 78.2mg/L), 5% acetic acid (pH 2.68), 0.004M hydrochloric acid (pH 2.71), 0.01M hydrochloric acid chlorine
Sour sodium (50 μ L sodium hypochlorite are added in 100mL 0.01M hydrochloric acid) and sodium hypochlorite (the 100mL10% second of 10% acetic acid+0.05%
50 μ L sodium hypochlorite are added in acid) at room temperature standing effect 15min, stainless steel substrates are transferred in 10mL nertralizers acted on rapidly
10min, then rinse stainless steel substrates 3 times with sterile PBS (0.05M, pH 7.2), add 10mL sterile PBS (0.05M, pH
7.2) 15min (250W), is ultrasonically treated, plate count, parallel test 3 times are carried out to bacteria suspension.
The acidic electrolytic water of comparative study embodiment 1 and with the various chemical modification water of its pH, ACC, ORP identical to waxy
The killing effect (being shown in Table 2) of bacillus biofilm.
Table 2 contrasts the killing effect of acidic electrolytic water and chemical modification water to bacillus cereus biofilm
Note：With a line, different letters represent significant difference (P ＜ 0.05), similarly hereinafter.
Interpretation of result：Sodium hypochlorite (sequence number 3) is markedly less than identical to the killing effect of bacillus cereus biofilm
ACC acidic electrolytic water.After sodium hypochlorite effect 1min, the viable count of biofilm only have dropped 1.13log CFU/cm2, make
After 15min, biofilm viable count is 3.35log CFU/cm2, it is more residual than on biofilm after acidic electrolytic water effect 1min
The viable count deposited is more.This is consistent with Liu etc. result of study, and under identical ACC, the germicidal efficiency of acidic electrolytic water is far above
Conventional chloride bactericide (Liu et al, 2006).Acetic acid (sequence number 4) and hydrochloric acid (sequence number 5) are to bacillus cereus biofilm
Acidic electrolytic water of the killing effect substantially than identical pH it is poor.For example, biofilm through acetic acid and hydrochloric acid effect 1min after, viable bacteria
Number have dropped 0.96log CFU/cm respectively2With 1.09log CFU/cm2；With the extension of action time, biofilm viable count
Slightly reduce, acetic acid and hydrochloric acid effect 15min after, remaining biofilm viable count is respectively on stainless steel substrates
3.19logCFU/cm2With 3.34log CFU/cm2, sterilizing rate is respectively 96.6% and 96.2%, is acted on than acidic electrolytic water
1min killing rate is also low.Hydrochloric acid and sodium hypochlorite mixed solution (sequence number 6) or acetic acid and sodium hypochlorite mixed solution (sequence number 7)
With the ORP suitable with acidic electrolytic water, but these chemical modification water are bright to the killing effect of bacillus cereus biofilm
It is aobvious to be weaker than acidic electrolytic water.Biofilm acts on through hydrochloric acid and sodium hypochlorite mixed solution or acetic acid with sodium hypochlorite mixed solution
After 1min, viable count have dropped 0.94log CFU/cm respectively2With 0.96log CFU/cm2, sterilizing rate is respectively 88.0% He
88.5%；Biofilm is residual after hydrochloric acid and sodium hypochlorite mixed solution or acetic acid act on 15min with sodium hypochlorite mixed solution
The viable count deposited is respectively 3.21log CFU/cm2With 2.92log CFU/cm2。
2nd, ACC kills the influence of bacillus cereus biofilm to acidic electrolytic water
Acidic electrolytic water a, b, c of the acidic electrolytic water of embodiment 1 and test example 1 are analyzed and (are shown in Table 3)
The ACC of table 3 kills the influence of bacillus cereus biofilm to acidic electrolytic water
3rd, pH kills the influence of bacillus cereus biofilm to acidic electrolytic water
Acidic electrolytic water d, e, f of the acidic electrolytic water of embodiment 1 and test example 2 are analyzed and (are shown in Table 4)
The pH of table 4 kills the influence of bacillus cereus biofilm to acidic electrolytic water
4th, ORP kills the influence of bacillus cereus biofilm to acidic electrolytic water
Acidic electrolytic water g, h of the acidic electrolytic water of embodiment 1 and test example 3 are analyzed and (are shown in Table 5)
The ORP of table 5 kills the influence of bacillus cereus biofilm to acidic electrolytic water
Conclusion：
(1) acidic electrolytic water of the invention can not only kill viable bacteria, and be low pH (2.7 under given conditions
~3.0), can be to waxy gemma bar, under high ACC (55~80mg/L) and high ORP (1150~1180mV) three synergy
The food-borne pathogens biofilm of this kind of better resistance of bacterium has very strong killing effect, and it kills the effect of biofilm
Mechanism is not low pH, the result of high ACC or high ORP independent roles, but the result of three's synergy；
2) preparation method of the invention is simple, cost is relatively low, is readily produced, lucifuge, 35 DEG C and under confined conditions, storage
When depositing no more than 9 days, there is preferable storage stability, the influence to pH, oxidation-reduction potential and higher effective chlorine density value
It is very small and the effect of can still keep stronger killing food-borne pathogens biofilm；
3) the invention provides the new application that acidic electrolytic water kills food-borne pathogens biofilm, killed available for preparation
Go out pharmaceutical composition, food additives composition and the disinfectant or cleaning agent of food-borne pathogens biofilm, is food
Control, the removing of biofilm provide new approach in industry, and then provide reliable guarantee for the security of food.
Finally it should be noted that above example is merely to illustrate technical scheme rather than the present invention is protected
The limitation of scope, although being explained in detail with reference to preferred embodiment to the present invention, one of ordinary skill in the art should manage
Solution, can modify or equivalent substitution to technical scheme, without departing from technical solution of the present invention essence and
Scope.
Claims (5)
1. acidic electrolytic water is used to prepare the pharmaceutical composition for killing food-borne pathogens biofilm or food additives combination
The new application of thing, the preparation method of described acidic electrolytic water, comprises the following steps：
(1) it is electrolysed：
In a cell, it is 0.05~0.1% chloro electrolyte solution as electrolyte using mass concentration, electrolysis 10~
15min, obtained in anode region pH value be 2.7~2.8, effective chlorine density is 55~80mg/L, oxidation-reduction potential be 1150~
1180mV acidic electrolytic water；In electrolytic process, the effective chlorine per 5min using indirect iodometric processes measure anode region electrolysis water is dense
Degree, pH meter measure pH value, oxidation-reduction potential test pen measure oxidation-reduction potential；
(2) lucifuge storage is sealed：
Acidic electrolytic water prepared by step (1), under the conditions of lucifuge, sealed storage in 35 DEG C of incubators is positioned over, during storage
Between be no more than 9 days.
2. acidic electrolytic water according to claim 1 is used to prepare the drug regimen for killing food-borne pathogens biofilm
The new application of thing or food additives composition, it is characterised in that：The acidic electrolytic water is preparing killing food-borne pathogens
Application in the disinfectant or cleaning agent of biofilm.
3. acidic electrolytic water according to claim 1 is used to prepare the drug regimen for killing food-borne pathogens biofilm
The new application of thing or food additives composition, it is characterised in that：The effective chlorine density of the acidic electrolytic water is 70~80mg/
L。
4. acidic electrolytic water according to claim 1 is used to prepare the drug regimen for killing food-borne pathogens biofilm
The new application of thing or food additives composition, it is characterised in that：The oxidation-reduction potential of the acidic electrolytic water be 1170~
1180mV。
5. acidic electrolytic water according to claim 1 is used to prepare the drug regimen for killing food-borne pathogens biofilm
The new application of thing or food additives composition, it is characterised in that：The chloro electrolyte is that mass concentration is 0.05~0.1%
Sodium chloride.
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| CN109722352B (en) * | 2018-12-21 | 2021-02-12 | 中国农业大学 | Combined cleaning agent, method and system for removing biofilm |
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| CN110583646A (en) * | 2019-09-19 | 2019-12-20 | 江西农业大学 | Foam type subacid electrolyzed water disinfectant for chicken farm |
| CN111534387A (en) * | 2020-05-20 | 2020-08-14 | 李树松 | Cleaning agent and preparation method and application thereof |
| CN114097818A (en) * | 2020-08-25 | 2022-03-01 | 张宝生 | High-efficiency disinfectant and preparation method thereof |
| CN112806519A (en) * | 2021-01-15 | 2021-05-18 | 上海海洋大学 | Method for efficiently removing bacterial biofilm |
| CN113670904A (en) * | 2021-07-27 | 2021-11-19 | 仲恺农业工程学院 | Application of acidic electrolyzed water in preparation of preparation for removing residual antibiotic norfloxacin in food |
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