CN113104929A - Non-thermal sterilization method for high-frequency ultrasonic synergistic plasma activated water treatment - Google Patents
Non-thermal sterilization method for high-frequency ultrasonic synergistic plasma activated water treatment Download PDFInfo
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- CN113104929A CN113104929A CN202110486995.7A CN202110486995A CN113104929A CN 113104929 A CN113104929 A CN 113104929A CN 202110486995 A CN202110486995 A CN 202110486995A CN 113104929 A CN113104929 A CN 113104929A
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/30—Treatment of water, waste water, or sewage by irradiation
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/34—Treatment of water, waste water, or sewage with mechanical oscillations
- C02F1/36—Treatment of water, waste water, or sewage with mechanical oscillations ultrasonic vibrations
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2303/00—Specific treatment goals
- C02F2303/04—Disinfection
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Abstract
The invention discloses a non-thermal sterilization method for high-frequency ultrasonic synergistic plasma activated water treatment, which comprises the following steps: treating the sterile water by using a plasma generating device to obtain plasma activated water; mixing the object to be treated with plasma activated water, and then starting high-frequency ultrasonic treatment; the high-frequency ultrasonic treatment frequency is 581-1144 kHz, the high-frequency ultrasonic treatment power is 50-200W, the treatment time is 5-15 min, and the treatment temperature is 20-25 ℃. The invention combines ultrasonic and plasma activated water, effectively avoids the problem of drug residue after sterilization, and can achieve better sterilization effect in a shorter time.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a method for sterilizing water by activating plasmas through high-frequency ultrasound.
Background
Coli is a gram-negative bacillus, mostly exists in single or two forms, has no spores, mostly has capsular or microcapsular structures, has pili, and has flagella. Most strains of E.coli are harmless, but a small fraction of opportunistic pathogens can produce powerful toxins under specific conditions causing serious diseases in humans and animals. For example, in raw meat products, raw milk and vegetables polluted by excrement, a plurality of shiga toxin-producing escherichia coli, in particular escherichia coli O157: H7 exist, the shiga toxin produced by the shiga toxin can cause symptoms such as nausea, vomiting, enteritis, severe abdominal pain and bloody diarrhea, and in some cases, symptoms such as renal failure, hemolytic uremic syndrome and the like can also occur, and even death can be caused in severe cases. The Escherichia coli O157H 7 has strong viability in the external environment and can survive for 9 months in the minced beef at the temperature of minus 20 ℃; the bacterium has strong resistance to acid, and can survive for more than 56 days under the condition that the pH is 4.0.
Consumers have increasingly demanded food products, and non-heat sterilization techniques have received increasing attention because they largely retain the original flavor and nutrition of food products. In recent years, plasma activated water has shown an advantage in sterilization, and is rich in active substances such as active oxygen and active nitrogen, and the active oxygen can act on peptidoglycan in cell walls, fatty acid in lipid bilayers and phospholipid bilayers of cell membranes, so that peptide bonds are cleaved and amino acid side chains are oxidized, which leads to cell wall rupture, cell membrane permeability change and even rupture, and finally death; the active nitrogen can inhibit the generation of ATP and enzyme activity, influence the metabolic pathway of iron ions to cause cell damage, and finally cause cell death.
The main mechanism of ultrasonic sterilization is known as the cavitation effect. The cavitation process generates tiny cavitation bubbles, the cavitation bubbles are broken to form a plurality of new small bubbles, local high temperature and high pressure, microflow, microjet and free radicals are generated, the free radicals can erode cell wall structures, and the actions jointly cause mechanical and chemical damage of cells, and finally cause cell inactivation. Mechanical effects also occur in the whole process of ultrasonic treatment, the transmission process of ultrasonic waves can cause the movement of liquid media, and irregular mutual collision and extrusion among molecules can generate shear stress, turbulence and the like, which can make cell walls thinner and more susceptible to the interference of external factors.
Invention 201710987312.X non-thermal sterilization method combining low temperature plasma and ultrasonic treatment teaches the following: firstly, treating staphylococcus aureus by dielectric barrier discharge low-temperature plasma, and then carrying out low-frequency ultrasonic treatment; the low-temperature plasma used in this method needs to be in direct contact with the object to be treated, since the plasma is generated by ionizing a gas at a high energy, heat accumulation is inevitably generated when treating a sample, and a sterilization treatment operation must be performed on the apparatus, which imposes a large limitation on the means of treating the sample.
Disclosure of Invention
The invention aims to provide a non-thermal sterilization method for activating water by using high-frequency ultrasonic synergistic plasma, which is used for killing Escherichia coli O157: H7 and reducing the biological hazard thereof and has the characteristics of good sterilization effect and environmental friendliness.
In order to solve the technical problem, the invention provides a non-thermal sterilization method for high-frequency ultrasonic synergistic plasma activated water treatment, which comprises the following steps:
1) treating the sterile water by using a plasma generating device to obtain plasma activated water;
2) performing high-frequency ultrasonic treatment (high-frequency ultrasonic synergistic plasma activated water treatment) by using the plasma activated water:
mixing the object to be treated with plasma activated water, and then starting (immediately starting) high-frequency ultrasonic treatment;
the high-frequency ultrasonic treatment frequency is 581-1144 kHz, the high-frequency ultrasonic treatment power is 50-200W, the treatment time is 5-15 min, and the treatment temperature is 20-25 ℃.
As an improvement of the non-thermal sterilization method, the step 1) is as follows:
placing the sterile water under a spray head of a plasma generating device (plasma surface treatment equipment), starting a high-voltage radio frequency generator, taking air as an air source, and generating plasma by a central electrode to treat the sterile water so as to obtain plasma activated water;
the preparation power is 650W-850W, the volume of the sterile water is 30-100 ml, the distance between the injection port of the plasma generating device and the surface of the sterile water is 6-9 cm, the treatment time is 30-90 s, and the preparation gas source is air.
As a further improvement of the non-thermal sterilization method of the present invention, in the preparation of the plasma-activated water of step 1): the preparation power is 650W, the distance between the injection port of the plasma generation device and the surface of the sterile water is 6-9 cm, the preparation time is 60s, the volume of the sterile water is 50mL, and the preparation gas source is air.
As a further improvement of the non-thermal sterilization method of the present invention, in the step 2), the object to be processed: mixing the plasma activated water in a volume ratio of 1: 5-9.
As a further improvement of the non-thermal sterilization method, in the step 2), the high-frequency ultrasonic treatment frequency is 581kHz, the high-frequency ultrasonic treatment power is 167W, the plasma activated water and the bacterial liquid are mixed according to the volume ratio of 5-9: 1, the treatment time is 5-15 min, and the treatment temperature is 25 ℃.
In the present invention, the samples before and after the treatment were subjected to colony counting.
Compared with plasma, the plasma activated water has the advantages of simple preparation process, relatively low cost, capability of being prepared at any time, and no problems of transportation, storage and preservation. But the service life of free radicals in the plasma activated water is short, and the sterilization effect of the plasma activated water is limited to a certain extent, so that the problem of drug residue after sterilization is effectively avoided by combining the ultrasonic and the plasma activated water, and meanwhile, a better sterilization effect can be achieved in a short time.
Compared with the prior art, the invention has the advantages that:
(1) reducing the dosage of plasma activated water;
(2) the invention adopts the combination of high-frequency ultrasound and plasma activated water to treat the escherichia coli, and has very good synergistic sterilization effect. Compared with single treatment, the combination of the two technologies can achieve the sterilization effect which can be achieved only under the severe single treatment condition under the milder condition, can reduce the sterilization time, and plays the roles of reducing energy consumption and saving resources.
(3) The invention adopts the synergistic effect of high-frequency ultrasound and plasma activated water, and both the two technologies are non-heat treatment technologies, thereby having great application prospect in the aspect of sterilization treatment of heat-sensitive materials.
In conclusion, the method greatly improves the killing efficiency of the escherichia coli, enhances the effectiveness of the ultrasonic sterilization process and is beneficial to expanding the industrial application of the ultrasonic sterilization.
Detailed Description
The invention will be further described with reference to specific examples, but the scope of the invention is not limited thereto.
Examples 1,
Step one, preparation of bacterial suspension (this is conventional technology):
coli O157H 7 standard strain ATCC 35150 was inoculated into a common broth and cultured in a shaker at 37 ℃ for about 17 hours to a stationary phase. Taking a proper amount of bacterial liquid, centrifuging, removing supernatant, suspending in sterile physiological saline with the mass fraction of 0.85%, and repeating the operation for 1-2 times. Diluting the resuspended bacterial liquid to obtain a concentration of about 1 × 108CFU/mL of bacterial suspension, stored at 4 ℃ until use.
Step two, preparation of plasma activated water:
the method is characterized in that plasma activated water is prepared by a plasma generating device (namely, plasma surface treatment equipment, Shenzhen Dongxing high-tech automated equipment, Inc.), sterile water is used as an original material, and the preparation method is carried out at normal temperature and comprises the following specific preparation steps:
50mL of sterile water was added to a 150mL beaker placed just below the plasma jet head with the liquid level 8cm from the jet head, and the beaker was placed in an ice-water bath to maintain the temperature. The treatment power is 650W, the treatment time is 30s, air is used as discharge gas, the pressure of the discharge gas is 0.2MPa, and the flow rate of the discharge gas is 39L/min; obtaining the plasma activated water.
Step three, activating water treatment by ultrasonic and plasma:
and (3) adding 1mL of the bacterial suspension obtained in the first step into 9mL of the plasma activated water obtained in the second step, immediately putting the bacterial suspension into an ultrasonic reactor for ultrasonic treatment (Meinhardt ultraschalaltechnik technology import company, Middy, China, and China), controlling the ultrasonic frequency to be 581kHz, the ultrasonic power to be 167W and the ultrasonic time to be 15min, and externally connecting a circulating water bath to control the temperature in the ultrasonic reactor to be 25 ℃ to obtain the treated bacterial liquid.
Step four, determining the total number of colonies
Diluting 100 μ L of the treated bacterial liquid with 0.85% sterile physiological saline gradient to appropriate concentration, taking 100 μ L of the diluted liquid, coating on a plate counting agar culture medium, culturing at 37 deg.C for 17h, and counting the number of colonies. And (4) diluting the untreated bacterial suspension (obtained in the step one) to the same multiple to obtain a blank control, wherein the bacterial reduction is the difference between the total number of the colonies in the control group and the total number of the colonies in the treatment group.
Experiment I,
The following 2 comparative treatment groups were set:
the comparative treatment group 1 was subjected to single ultrasonic treatment at room temperature, specifically as follows:
the second step of the example 1 is eliminated, and the plasma activated water in the third step of the example 1 is changed into sterile water; the rest is equivalent to embodiment 1.
The comparative treatment group 2 was treated with water by single plasma activation at room temperature as follows:
the sonication in step three of example 1 was eliminated by adding plasma-activated water to the bacterial suspension and mixing, followed by standing for 15 minutes, and the rest was identical to example 1.
The results show that the comparative treatment group 1 (the escherichia coli bacterial suspension is subjected to single ultrasonic treatment for 15min at normal temperature) can inactivate 0.585log CFU/mL, the comparative treatment group 2 (the escherichia coli bacterial suspension is subjected to single plasma activation water treatment for 15min at normal temperature) can inactivate 0.353log CFU/mL, and the example 1 (the escherichia coli bacterial suspension is subjected to ultrasonic synergistic plasma activation water treatment for 15min) can inactivate 6.055log CFU/mL. Therefore, the method has a good effect of inactivating escherichia coli under the treatment of ultrasonic synergistic plasma activated water.
Example 2, the treatment time in the preparation of plasma activated water in the second step of example 1 was changed from 30s to 60 s. The rest is equivalent to embodiment 1.
Also as with the set-up of experiment one, the corresponding 2 control treatment groups were set up.
The results show that the comparative treatment group 1 (the escherichia coli bacterial suspension is subjected to single ultrasonic treatment for 15min at normal temperature) can inactivate 0.585log CFU/mL, the comparative treatment group 2 (the escherichia coli bacterial suspension is subjected to single plasma activation water treatment for 15min at normal temperature) can inactivate 0.820log CFU/mL, and the example 2 (the escherichia coli bacterial suspension is subjected to ultrasonic synergistic plasma activation water treatment for 15min) can inactivate 8.406log CFU/mL. Therefore, example 2 has a better sterilization effect than example 1.
Example 3-1, the ultrasonic treatment frequency in the synergistic treatment in example 2 was changed from 581kHz to 861kHz, and the rest was equivalent to example 2.
Escherichia coli suspension treated by ultrasonic synergistic plasma activated water can be inactivated by 3.64log CFU/mL after 15 min.
Example 3-2, the ultrasonic treatment frequency in the synergistic treatment in example 2 was changed from 581kHz to 1144kHz, and the rest was equivalent to example 2.
Escherichia coli suspension treated by ultrasonic synergistic plasma activated water can be inactivated by 0.96log CFU/mL after 15 min.
In example 4, the ultrasonic processing power in the synergistic processing in example 2 was changed from 167W to 133W, and the rest was the same as that in example 2.
Escherichia coli suspension treated by ultrasonic synergistic plasma activated water can be inactivated by 3.66log CFU/mL after 15 min.
In comparative example 1-1, the temperature in the external circulating water bath controlled ultrasonic reactor in example 2 was changed from 25 ℃ to 15 ℃, and the rest was the same as example 2.
After the Escherichia coli suspension subjected to ultrasonic synergistic plasma activated water treatment is inactivated for 15min, 4.23log CFU/mL can be inactivated, namely, the sterilization effect is not as good as that of example 2.
Comparative examples 1-2, the temperature in the external circulating water bath controlled ultrasonic reactor in example 2 was changed from 25 ℃ to 35 ℃, and the rest was the same as example 2.
The inactivation quantity of the escherichia coli bacterial suspension subjected to the ultrasonic synergistic plasma activation water treatment is no obvious difference from that of the escherichia coli bacterial suspension treated in the example 2 after 15min, and energy used for heating is consumed instead.
Finally, it is also noted that the above-mentioned lists merely illustrate a few specific embodiments of the invention. It is obvious that the invention is not limited to the above embodiments, but that many variations are possible. All modifications which can be derived or suggested by a person skilled in the art from the disclosure of the present invention are to be considered within the scope of the invention.
Claims (5)
1. The non-thermal sterilization method for activating water treatment by high-frequency ultrasonic synergistic plasma is characterized by comprising the following steps of:
1) treating the sterile water by using a plasma generating device to obtain plasma activated water;
2) and performing high-frequency ultrasonic treatment by using the plasma activated water:
mixing the object to be treated with plasma activated water, and then starting high-frequency ultrasonic treatment;
the high-frequency ultrasonic treatment frequency is 581-1144 kHz, the high-frequency ultrasonic treatment power is 50-200W, the treatment time is 5-15 min, and the treatment temperature is 20-25 ℃.
2. The non-thermal sterilization method according to claim 1, wherein the step 1) is:
placing the sterile water under a spray head of a plasma generating device, starting a high-pressure radio frequency generator, and treating the sterile water by using air as an air source and generating plasma by a central electrode so as to obtain plasma activated water;
the preparation power is 650W-850W, the volume of the sterile water is 30-100 ml, the distance between the injection port of the plasma generating device and the surface of the sterile water is 6-9 cm, the treatment time is 30-90 s, and the preparation gas source is air.
3. The non-thermal sterilization method as set forth in claim 2, wherein the plasma-activated water of step 1) is prepared by: the preparation power is 650W, the distance between the injection port of the plasma generation device and the surface of the sterile water is 6-9 cm, the preparation time is 60s, the volume of the sterile water is 50mL, and the preparation gas source is air.
4. The non-thermal sterilization method according to any one of claims 1 to 3, characterized in that:
in the step 2), the object to be processed: mixing the plasma activated water in a volume ratio of 1: 5-9.
5. The non-thermal sterilization method according to claim 4, wherein:
in the step 2), the high-frequency ultrasonic treatment frequency is 581kHz, the high-frequency ultrasonic treatment power is 167W, the plasma activated water and the bacterial liquid are mixed according to the volume ratio of 5-9: 1, the treatment time is 5-15 min, and the treatment temperature is 25 ℃.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114847336A (en) * | 2022-04-28 | 2022-08-05 | 江苏省农业科学院 | Plasma activated water and ultrasonic wave sterilization equipment applied to crayfishes |
| CN117461726A (en) * | 2023-09-17 | 2024-01-30 | 郑州轻工业大学 | Method for removing alternaria alternata toxin and bitter substances in citrus slag |
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| CN107853538A (en) * | 2017-10-20 | 2018-03-30 | 浙江大学 | A kind of non-thermal sterilization method of low temperature plasma combining ultrasonic processing |
| CN110913553A (en) * | 2019-12-12 | 2020-03-24 | 大连理工大学 | High-efficient plasma activated water produces equipment |
| CN110897076A (en) * | 2019-11-07 | 2020-03-24 | 华南理工大学 | Method for cleaning and preserving fruits and vegetables based on ultrasonic-assisted plasma activated water |
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- 2021-05-04 CN CN202110486995.7A patent/CN113104929A/en active Pending
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| CN107853538A (en) * | 2017-10-20 | 2018-03-30 | 浙江大学 | A kind of non-thermal sterilization method of low temperature plasma combining ultrasonic processing |
| CN110897076A (en) * | 2019-11-07 | 2020-03-24 | 华南理工大学 | Method for cleaning and preserving fruits and vegetables based on ultrasonic-assisted plasma activated water |
| CN110913553A (en) * | 2019-12-12 | 2020-03-24 | 大连理工大学 | High-efficient plasma activated water produces equipment |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114847336A (en) * | 2022-04-28 | 2022-08-05 | 江苏省农业科学院 | Plasma activated water and ultrasonic wave sterilization equipment applied to crayfishes |
| CN117461726A (en) * | 2023-09-17 | 2024-01-30 | 郑州轻工业大学 | Method for removing alternaria alternata toxin and bitter substances in citrus slag |
| CN117461726B (en) * | 2023-09-17 | 2024-08-30 | 郑州轻工业大学 | Method for removing alternaria alternata toxin and bitter substances in citrus slag |
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