CN108651811B - Method for enhancing sterilization effect by optimizing ultrahigh pressure boosting process - Google Patents
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- 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
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- A23L3/015—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with pressure variation, shock, acceleration or shear stress or cavitation
- A23L3/0155—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with pressure variation, shock, acceleration or shear stress or cavitation using sub- or super-atmospheric pressures, or pressure variations transmitted by a liquid or gas
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Abstract
The invention discloses a method for strengthening a sterilization effect by optimizing an ultrahigh pressure boosting process. The method comprises the following steps: and (3) putting the food into an ultrahigh pressure device, increasing the pressure to a target pressure in a continuous linear pressure increasing mode, maintaining the pressure, and then releasing the pressure, so that the sterilization of the food can be realized. The boosting rate of the continuous linear boosting is 1-10 MPa/s. Compared with the conventional ultrahigh pressure sterilization method (such as a slow and stepped pressure boosting process), the method disclosed by the invention can achieve the effect of strengthening sterilization only under the condition of adjusting the self pressure boosting parameters of the equipment, realizes the purposes of reducing the pressure level, shortening the sterilization time, reducing the energy consumption of ultrahigh pressure treatment and improving the working efficiency, has important practical production significance, and has strong guiding significance on the design and production of a supercharger in the ultrahigh pressure equipment.
Description
Technical Field
The invention belongs to the field of food safety, and particularly relates to a method for enhancing a sterilization effect by optimizing an ultrahigh pressure boosting process.
Background
The ultrahigh pressure technology is a new non-thermal processing technology, can ensure good sterilization effect and simultaneously maintain the natural color, fragrance, taste, texture, nutrition and other qualities of food, and is widely applied to the field of food processing. The ultra-high pressure sterilization is a very important link for ensuring the safety and the quality of food, and the application and the popularization of the ultra-high pressure sterilization in food processing are restricted due to the insufficient ultra-high pressure sterilization effect. How to strengthen the ultrahigh pressure sterilization effect on the premise of reducing the pressure level and shortening the processing time so as to reduce the energy consumption of ultrahigh pressure processing and improve the working efficiency is a problem which needs to be solved urgently and has great practical application significance. At present, it is consistently considered that the factors influencing the ultra-high pressure sterilization effect mainly include treatment temperature, pressure magnitude, pressure holding time, types and characteristics of target microorganisms, sample properties and the like, and the existing strengthening method is realized on the premise of changing the several influencing factors, such as increasing temperature, time and the like. If the sterilization effect can be increased on the premise of not increasing the treatment temperature, the treatment pressure, the treatment time and the product properties, the efficiency and the actual operability of the ultrahigh pressure processing can be greatly improved, and the method is very important for accelerating the popularization of the ultrahigh pressure technology.
The main parameters of the ultrahigh pressure boosting process are the boosting rate and the boosting mode. The pressure increasing rate is the speed of the pressure increasing process, namely the time (unit is MPa/min or MPa/s) required for reaching the working pressure. The boosting method is classified into a stepwise method and a linear method. In the conventional ultrahigh pressure sterilization method, the boosting speed is slow in the boosting process, the boosting mode is stepped, and the sterilization effect is limited.
Disclosure of Invention
The invention aims to provide a method for enhancing the sterilization effect by optimizing the boosting process of ultrahigh pressure, which realizes the purpose of enhancing the sterilization effect of ultrahigh pressure by optimizing the boosting process (boosting mode and boosting speed).
The invention aims to provide an ultrahigh pressure sterilization method, which comprises the following steps: and (3) putting the food into an ultrahigh pressure device, increasing the pressure to a target pressure in a continuous linear pressure increasing mode, maintaining the pressure, and then releasing the pressure, so that the sterilization of the food can be realized.
In the above method, the continuous linear pressure increase means that the pressure increase is continuous and uninterrupted in the pressure increase process, and the pressure is proportional to the time.
In the method, the continuous linear boosting can be realized by adjusting the working sequence of the superchargers in the ultrahigh-pressure equipment. The boosting mode is divided into a step type and a linear type and is determined by different types of superchargers, and the ultrahigh-pressure superchargers are divided into a one-way supercharger and a two-way supercharger. The one-way supercharger can only generate limited high-pressure medium in the process of primary supercharging, and after the supercharging is finished, the pressure transfer medium must be supplemented through reversing, and then the supercharging is continued. This results in a period of boost lag, which appears as a plateau in the boost curve. Because a boosting process comprises a plurality of stagnation periods, a plurality of platforms are included in the boosting process, and the boosting curve is in a step shape. And the bidirectional supercharger can continue supercharging by reversing after primary supercharging is finished, so that a stagnation period can not occur, and a boosting curve is expressed as a linear type. At present, a large-scale ultrahigh-pressure device usually uses several one-way superchargers or two-way superchargers at the same time, so that different supercharging processes (supercharging speed and mode) can be generated by combining different superchargers.
In the method, the pressure increasing rate of the continuous linear pressure increasing can be 1-10 MPa/s, such as 2-10 MPa/s, 2-5 MPa/s, 5-10 MPa/s, 1MPa/s, 2MPa/s, 5MPa/s or 10 MPa/s.
In the method, the pressure relief time is not more than 3 s.
In the above method, specifically, different sterilization conditions may be selected depending on the type of target microorganism and/or food:
the target microorganism is escherichia coli; the sterilization conditions were as follows:
the target pressure is 250-400 MPa, and specifically can be 250-300 MPa, 250MPa or 300 MPa;
the pressure maintaining time is 5-10 min, specifically 5 min;
the pressure maintaining temperature is 15-30 ℃, and specifically can be 25 ℃;
the pressure transmission medium is water.
The food is purple sweet potato juice; the sterilization conditions were as follows:
the target pressure is 400-600 MPa, and specifically can be 400MPa, 500MPa or 600 MPa;
the pressure maintaining time is 20-30 min, specifically 25 min;
the pressure maintaining temperature is 15-30 ℃, and specifically can be 25 ℃;
the pressure transmission medium is water.
The target microorganism is staphylococcus aureus; the sterilization conditions were as follows:
the target pressure is 300-400 MPa, and specifically can be 400 MPa;
the pressure maintaining time is 5-20 min, specifically 5 min;
the pressure maintaining temperature is 15-30 ℃, and specifically can be 25 ℃;
the pressure transmission medium is water.
The invention has the following beneficial effects:
compared with the conventional ultrahigh pressure sterilization method (such as a slow and stepped pressure boosting process), the method disclosed by the invention can achieve the effect of strengthening sterilization only under the condition of adjusting the self pressure boosting parameters of the equipment, realizes the purposes of reducing the pressure level, shortening the sterilization time, reducing the energy consumption of ultrahigh pressure treatment and improving the working efficiency, has important practical production significance, and has strong guiding significance on the design and production of a supercharger in the ultrahigh pressure equipment.
Drawings
Fig. 1 is a graph of different boosting processes in embodiment 1, wherein fig. 1(a) is a linear fast, fig. 1(B) is a linear slow, fig. 1(C) is a step fast, and fig. 1(D) is a step slow.
FIG. 2 shows the killing effect of different boosting processes on Escherichia coli under ultra-high pressure (300MPa/5min) in example 1.
Fig. 3 is a graph illustrating different boosting processes in embodiment 2 and embodiment 3, wherein fig. 3(a) is a mode a, stepwise slow; FIG. 3(B) is a B mode, step fast; fig. 3(C) shows a C mode, which is linear fast.
FIG. 4 shows the killing effect of different boosting processes on Escherichia coli and Staphylococcus aureus in example 3.
Detailed Description
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Example 1, for E.coli O157: h7 performing ultra-high pressure sterilization
The strain used in the experiment is E.coli O157: H7NCTC12900, can not produce Shiga toxin stx1 and stx2, belongs to a detoxification type strain and is derived from a British national standard strain collection; TSA, TSB medium involved in the experiment was purchased from Oboxing Biotechnology, Beijing; the ultra high pressure processing apparatus is available from Stansted, UK under the product model number FPG 7100: 9/2C.
Different boosting modes are adopted for the Escherichia coli O157: h7 is sterilized under ultrahigh pressure and compared with the sterilization effect, and the specific steps are as follows:
(1) escherichia coli O157: activation and preparation of H7 thallus
Streaking the strain frozen at-80 deg.C on solid TSA culture dish, culturing at 37 deg.C overnight (12h), picking single colony in liquid TSB culture medium, culturing at 37 deg.C with shaking table at 200rpm overnight (12h), transferring 1:100 into fresh liquid TSB culture medium, culturing at 37 deg.C with shaking table at 200rpm for 2.5h until OD600Filling into plastic bags for 0.8 sterile, and then performing ultra-high pressure sterilization.
(2) Optimization of boosting process
And setting the boosting rate of the equipment to be the fastest 10.0MPa/s and the slowest 1.0MPa/s according to the range of the parameters of the ultrahigh pressure device. And meanwhile, the working sequence of the supercharger is adjusted, so that the boosting modes are different: step-type, a stagnation period occurs in the boosting, namely, a plurality of platforms occur in the boosting process; linear-there is no lag phase in boosting, i.e. there is no platform in boosting. Different boosting rates and modes are combined, namely, four combined boosting processes of linear fast, linear slow, step fast and step slow are adopted. The graph of the four boosting processes is shown in fig. 1.
(3) Ultra-high pressure sterilization
The pressure holding level and the pressure holding time (the temperature can be correspondingly controlled according to the needs) are determined according to the properties of the pressed sample, and the pressure relief process system defaults to be generally less than 3 s.
Here, the temperature was set at room temperature (25 ℃) at 300MPa/5min (pressure maintenance at 300MPa for 5min), and the ratio of Escherichia coli O157 in the plastic bag was adjusted: h7 was subjected to sterilization treatment. The pressure transmission medium is water. The pressure relief time does not exceed 3 seconds.
(4) Plate counting judgment of sterilization effect
1ml of Escherichia coli O157 before and after the ultrahigh pressure treatment was taken: diluting H7 bacterial solution in 9mL of 0.85% physiological saline step by step, mixing 1mL of bacterial solution with about 15mL of liquid TSA culture medium in each dilution gradient, inverting after the plate is solidified, putting the plate into an incubator at 37 ℃ for culturing for 24-48H, and counting.
(5) Contrast of enhanced bactericidal effect
The bactericidal effect of the different boosting processes is shown in fig. 2, wherein N: number of Escherichia coli after ultrahigh pressure treatment, N0: original sourceThe number of E.coli cells, different letters in the figure, was significantly different (p)<0.05). By comparing the influence of the four boosting processes of linear high speed, linear low speed, step high speed and step low speed on the sterilization effect, the sterilization effect of the treatment with the high boosting rate is found to be better in the same boosting mode; under the same boosting rate, the linear boosting mode has better treatment and sterilization effects; namely, the sterilization effect of ultrahigh pressure can be enhanced by the rapid pressure increasing speed and the linear pressure increasing mode. Therefore, in practical application, the sterilization effect of the ultrahigh pressure can be enhanced by accelerating the pressure increase rate and adjusting the pressure increase rate to a linear pressure increase mode.
Example 2 ultra high pressure Sterilization of purple sweet Potato juice
Adopt different modes of stepping up to carry out the super high pressure to purple sweet potato juice and disinfect and compare bactericidal effect, concrete step is as follows:
(1) preparation of purple sweet potato juice
Cleaning purple sweet potato, peeling, cutting into 3-5mm slices, blanching in boiling water for 2-3min, cooling in cold water, draining, adding water at a ratio of 1:6(w/v), and pulping; adding white granulated sugar and food-grade lemon water into purple sweet potato juice to adjust sweet-acid ratio, grinding the purple sweet potato juice with the ingredients by a colloid mill, homogenizing in a high-pressure homogenizer under 30MPa, filling into a sterile 50mL PET bottle, sealing, and storing at 4 ℃ before ultra-high pressure sterilization.
(2) Setting of ultra-high pressure boosting parameter
Three different boosting processes are set: mode A, stepped slow speed (1 MPa/s); mode B, stepwise fast (2 MPa/s); mode C, linear fast (2MPa/s) (FIG. 3).
(3) Ultra high pressure sterilization
Setting target pressures of 400MPa, 500MPa and 600MPa at room temperature (25 ℃), maintaining the pressure for 25min, and then placing the purple sweet potato juice into an ultrahigh pressure cavity for sterilization. The pressure transmission medium is water. The pressure relief time does not exceed 3 s.
(4) Plate counting judgment of sterilization effect
The total number of bacterial colonies in the purple sweet potato juice after high-pressure treatment is measured, and the sterilization effect of the purple sweet potato juice under the ultrahigh pressure in different boosting processes is analyzed and compared. Diluting the purple sweet potato juice before and after the ultrahigh pressure treatment by ten-fold increasing gradient with 0.85% physiological saline, mixing 1mL of bacterial liquid with 15mL of flat counting culture medium in each dilution gradient, culturing at 37 ℃ for 48h after the flat is solidified, and counting, wherein the results are shown in table 1. It can be seen that the linear rapid boosting can achieve the best sterilization effect compared with other boosting processes under different pressure levels.
TABLE 1 Effect of different boosting procedures on the total number of colonies (-LogN/N) in purple sweet potato juice0)
EXAMPLE 3 ultra high pressure Sterilization of Escherichia coli and Staphylococcus aureus
(1) Activation and preparation of colibacillus and staphylococcus aureus thallus
The lyophilized powder of Escherichia coli E.coli O157: H7 (accession number: NCTC12900) and Staphylococcus aureus S.aureus (accession number: CGMCC.1861) was activated and stored at 4 ℃ on slant culture medium according to the respective instruction. Taking appropriate amount of tryptone soy and broth nutrient broth culture medium, inoculating the two strains into liquid culture medium with inoculating loop, respectively, and culturing at 37 deg.C for 2.5h (Escherichia coli) and 4h (Staphylococcus aureus). Centrifuging the cultured bacteria liquid at 4 deg.C and 8000rpm for 10min, discarding supernatant, and resuspending with sterile physiological saline to obtain bacteria liquid with final concentration of 107～108cfu/mL, sterile filling into plastic bags and then ultra-high pressure sterilization.
(2) Setting of ultra-high pressure boosting parameter
Three different boosting processes are set: mode A, stepped slow speed (1 MPa/s); mode B, stepwise fast (2 MPa/s); mode C, linear fast (2MPa/s) (FIG. 3).
(3) Ultra high pressure sterilization
Setting the target pressure for treating escherichia coli and staphylococcus aureus at room temperature (25 ℃) to be 250MPa and 400MPa respectively, maintaining the pressure for 5min, and sterilizing the bacterial liquid in the plastic package bag. The pressure transmission medium is water. The pressure relief time does not exceed 3 s.
(4) Plate counting judgment of sterilization effect
The bacterial colony number of the escherichia coli and the staphylococcus aureus after high-pressure treatment is measured, and the sterilization effect of the ultrahigh pressure in different pressure boosting processes is analyzed and compared. And (3) carrying out ten-fold incremental gradient dilution on the bacterial liquid before and after the ultrahigh pressure treatment by using 0.85% physiological saline, mixing 1mL of bacterial liquid with about 15mL of corresponding culture medium in each dilution gradient, respectively placing at proper temperature for culture, and counting. Wherein the total number of colonies is cultured at 37 deg.C for 48h, the total number of molds and yeasts is cultured at 28 deg.C for more than 72h, and Escherichia coli and Staphylococcus aureus are cultured at 37 deg.C for 24 h.
The results are shown in FIG. 4, where N: number of Escherichia coli after ultrahigh pressure treatment, N0: the number of original E.coli, different letters in the figure represent significant differences (p)<0.05). It can be seen that the mode C linear rapid boosting can achieve the best sterilization effect compared with other boosting processes under different pressure levels.
Claims (2)
1. An ultrahigh pressure sterilization method comprises the following steps: putting the food into an ultrahigh pressure device, increasing the pressure to a target pressure in a continuous linear pressure increasing mode, maintaining the pressure, and then releasing the pressure to realize the sterilization of the food;
the boosting rate of the continuous linear boosting is 2-10 MPa/s;
the target microorganism is escherichia coli; the sterilization conditions were as follows:
the target pressure is 250-400 MPa;
the pressure maintaining time is 5-10 min;
the pressure maintaining temperature is 15-30 ℃;
the pressure transmission medium is water;
the target microorganism is staphylococcus aureus; the sterilization conditions were as follows:
the target pressure is 300-400 MPa;
the pressure maintaining time is 5-20 min;
the pressure maintaining temperature is 15-30 ℃;
the pressure transmission medium is water.
2. An ultrahigh pressure sterilization method comprises the following steps: putting the food into an ultrahigh pressure device, increasing the pressure to a target pressure in a continuous linear pressure increasing mode, maintaining the pressure, and then releasing the pressure to realize the sterilization of the food;
the boosting rate of the continuous linear boosting is 2-10 MPa/s;
the food is purple sweet potato juice; the sterilization conditions were as follows:
the target pressure is 400-600 MPa;
the pressure maintaining time is 20-30 min;
the pressure maintaining temperature is 15-30 ℃;
the pressure transmission medium is water.
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