CN114403334A - Method for cold sterilization of fruit juice by curcumin-mediated photodynamic technology - Google Patents
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- 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 title claims abstract description 148
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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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/42—Preservation of non-alcoholic beverages
- A23L2/44—Preservation of non-alcoholic beverages by adding preservatives
-
- 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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/42—Preservation of non-alcoholic beverages
- A23L2/50—Preservation of non-alcoholic beverages by irradiation or electric treatment without heating
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
The invention relates to a food sterilization technology, and particularly discloses a method for cold sterilization of fruit juice by a curcumin-mediated photodynamic technology, which comprises the following steps: adding curcumin with different concentrations into sterile fruit juice, adding staphylococcus aureus suspension cultured to a stable stage, uniformly mixing at normal temperature in a dark place, and carrying out photodynamic treatment on the fruit juice to obtain the sterilized fruit juice. The invention adds common food additive curcumin as photosensitizer in fruit juice, and conducts mild cold sterilization by the mediation photodynamic technology. Under the sterilization mode, the highest sterilization rate of staphylococcus aureus in the fruit juice can reach more than 99.99 percent; compared with the traditional heat treatment, the method is convenient and fast, the treatment time is short, the change of sensory parameters in the fruit juice is reduced to a certain extent, and the method has great application potential in the food industry.
Description
Technical Field
The invention relates to the technical field of food sterilization, in particular to a method for cold sterilization of fruit juice by curcumin-mediated photodynamic technology.
Background
Sterilization in food is the most important operation link in the process of ensuring food safety. In recent years, efforts are underway at home and abroad to explore various advanced sterilization techniques to avoid harmful factors caused by sterilization and minimize the loss of nutrient elements in food. The development of modern food sterilization technology is evolving step by step, and the traditional heating sterilization mode is gradually changed into high-efficiency sterilization with heating time reduced as much as possible. In recent years, non-thermal sterilization technologies represented by cold sterilization technologies have appeared, including a series of novel sterilization technologies such as nano sterilization technology, vibration magnetic field sterilization technology, vacuum sterilization and the like, so as to keep the color, aroma, taste and the like of food as much as possible. With the improvement of living standard of people and the increasing demand of food taste, the sterilization technology of food is further explored and developed.
Curcumin is an active polyphenol substance extracted from rhizome of plant Curcuma rhizome, and has antitumor, antiinflammatory, blood sugar lowering, antioxidant, and apoptosis resisting pharmacological activities. Researches show that curcumin as a natural photosensitizer has strong killing effect on listeria monocytogenes, staphylococcus aureus, escherichia coli, salmonella and the like. In addition, the european food safety agency defines curcumin as a non-carcinogenic compound and authorizes its use as a food additive in the european union, which is also a food natural pigment permitted by the regulation of GB 2760-. Therefore, curcumin can be safely used in food.
Photodynamic sterilization is a technique that relies on the occurrence of non-thermal photophysical and photochemical reactions, which also require visible light and photosensitizers in the presence of oxygen. Wherein the visible light originates from a light emitting diode, LED, having a visible wavelength. Due to the characteristics of safety, environmental protection, low energy consumption and the like, the fresh-keeping technology for the food is more and more concerned in recent years. The existing research results show that individual or combined photosensitizer guided by 405nm and 460nm can effectively inactivate food-borne pathogenic bacteria in food, and the sterilization method can not only completely kill the bacteria, but also degrade toxic substances released by the bacteria, thereby avoiding secondary pollution. Therefore, the method is regarded as an ideal modern sterilization technology.
Disclosure of Invention
The invention aims to provide a method for cold sterilization of fruit juice by curcumin-mediated photodynamic technology, which can improve the defect of single light penetrability and avoid the damage of the traditional heat sterilization technology to the sensory parameters of the fruit juice to a certain extent by carrying out photodynamic treatment on the fruit juice for a period of time in the presence of photosensitizer curcumin.
In order to achieve the purpose, the invention is implemented according to the following technical scheme:
a method for cold sterilization of fruit juice by curcumin-mediated photodynamic technology comprises the following steps:
step one, adding curcumin with different concentrations into sterile fruit juice;
step two, adding staphylococcus aureus suspension cultured to a stable stage;
and step three, after uniformly mixing the juice at normal temperature in a dark place, carrying out photodynamic treatment on the juice to obtain the sterilized juice.
Further, the first step specifically includes:
1) preparing 4mmol/L curcumin DMSO solution, and diluting with normal saline to obtain 40 μmol/L curcumin solution containing 10% DMSO;
2) diluting the prepared curcumin solution with fruit juice to obtain sample solution with curcumin concentration of 20 μmol/L for use.
Further, the second step specifically includes:
1) selecting a staphylococcus aureus single colony in a TSB liquid culture medium, and performing shake culture at 37 ℃ for 12h to obtain a staphylococcus aureus suspension cultured to a stable stage;
2) diluting the obtained bacterial liquid with fruit juice, and mixing with the fruit juice obtained in step one to make the final bacterial concentration 106CFU/mL, curcumin concentration of 10 u mol/L.
Further, in the third step, the juice is uniformly mixed for 30 minutes in a dark place at normal temperature, and the juice is subjected to photodynamic treatment for 6-12 minutes.
Further, the photodynamic treatment conditions are as follows: a blue LED array was used with a power of 5W, a wavelength of 435-.
Compared with the traditional heat sterilization technology, the invention has the following advantages and beneficial effects:
1) the food additive curcumin is selected as a photosensitizer, and the mediated photodynamic technology can effectively kill staphylococcus aureus in fruit juice, thereby ensuring the use safety.
2) The method is efficient and simple, and the required cost is low.
3) Compared with the traditional heat treatment method, the method reduces the change of sensory parameters of the juice to a certain extent while sterilizing, and has wide application prospect in beverage industries such as juice and the like.
Drawings
Fig. 1 is a schematic diagram of curcumin-mediated photodynamic technical process.
FIG. 2 shows the bactericidal effect of curcumin-mediated photodynamic technology on Staphylococcus aureus in fruit juice under selected conditions.
Detailed Description
The present invention will be described in further detail with reference to examples for better describing the present invention, but the embodiments of the present invention are not limited thereto.
The test methods used in the following examples are all conventional methods unless otherwise specified; the materials, reagents and the like used are all commercially available reagents and materials unless otherwise specified.
The strains and materials used in the following examples:
staphylococcus aureus ATCC29213 was obtained from the China medical bacteria Collection. The used Tryptic Soy Agar (TSA) solid medium, Tryptic Soy Broth (TSB) liquid medium and Baird-Parker agar (BP) Staphylococcus aureus selection medium were purchased from Haibo Bio Inc. All media were prepared as indicated and autoclaved at 121 ℃ for 20 minutes before use.
Example 1
Preparing 4mmol/L curcumin DMSO solution, and diluting with normal saline to obtain 40 μmol/L curcumin solution containing 10% DMSO;
diluting the prepared curcumin solution with commercially available mango juice to obtain a sample solution with curcumin concentration of 20 mu mol/L for later use;
selecting a staphylococcus aureus single colony in a TSB liquid culture medium, and performing shake culture at 37 ℃ for 12h to obtain a staphylococcus aureus suspension cultured to a stationary phase;
centrifuging 1ml of the bacterial solution in a centrifuge at 4 deg.C and 8000rpm for 10 min, discarding supernatant, adding equal volume of physiological saline, resuspending, and diluting with commercially available mango juice to obtain final bacterial concentration of 2 × 106CFU/mL；
Mixing the diluted bacteria liquid with the fruit juice obtained in the step one to ensure that the concentration of the bacteria in the final sample is 106CFU/mL, the concentration of curcumin is 10 mu mol/L;
putting the sample solution into a sterile culture dish every 15mL, uniformly mixing for 30 minutes at normal temperature in a dark place, performing photodynamic treatment on the fruit juice for 6 minutes by using a blue light-emitting diode array with the power of 5W, the wavelength of 435-.
After the treatment, the sample solution was serially diluted with physiological saline, 0.1mL of the diluted solution was applied to a BP medium, and the number of staphylococcus aureus was counted. The results of the experiment are shown in FIG. 2.
Example 2
Preparing 4mmol/L curcumin DMSO solution, and diluting with normal saline to obtain 40 μmol/L curcumin solution containing 10% DMSO;
diluting the prepared curcumin solution with commercially available mango juice to obtain a sample solution with curcumin concentration of 20 mu mol/L for later use;
selecting a staphylococcus aureus single colony in a TSB liquid culture medium, and performing shake culture at 37 ℃ for 12h to obtain a staphylococcus aureus suspension cultured to a stationary phase;
centrifuging 1ml of the bacterial solution in a centrifuge at 4 deg.C and 8000rpm for 10 min, discarding supernatant, adding equal volume of physiological saline, resuspending, and diluting with commercially available mango juice to obtain final bacterial concentration of 2 × 106CFU/mL；
Mixing the diluted bacteria liquid with the fruit juice obtained in the step one to ensure that the concentration of the bacteria in the final sample is 106CFU/mL, the concentration of curcumin is 10 mu mol/L;
putting 15mL of sample solution into a sterile culture dish, uniformly mixing for 30 minutes at normal temperature in a dark place, performing photodynamic treatment on the fruit juice for 12 minutes by using a blue light-emitting diode array with the power of 5W, the wavelength of 435-.
After the treatment, the sample solution was serially diluted with physiological saline, 0.1mL of the diluted solution was applied to a BP medium, and the number of staphylococcus aureus was counted. The results of the experiment are shown in FIG. 2.
Example 3
Preparing 4mmol/L curcumin DMSO solution, and diluting with normal saline to obtain 40 μmol/L curcumin solution containing 10% DMSO;
diluting the prepared curcumin solution with commercially available pineapple juice to obtain a sample solution with curcumin concentration of 20 mu mol/L for later use;
selecting a staphylococcus aureus single colony in a TSB liquid culture medium, and performing shake culture at 37 ℃ for 12h to obtain a staphylococcus aureus suspension cultured to a stationary phase;
centrifuging 1ml of the bacterial solution in a centrifuge at 4 deg.C and 8000rpm for 10 min, discarding supernatant, adding equal volume of physiological saline, resuspending, and diluting with commercially available pineapple juice to obtain final bacterial concentration of 2 × 106CFU/mL；
Mixing the diluted bacteria liquid with the fruit juice obtained in the step one to ensure that the concentration of the bacteria in the final sample is 106CFU/mL, the concentration of curcumin is 10 mu mol/L;
putting the sample solution into a sterile culture dish every 15mL, uniformly mixing for 30 minutes at normal temperature in a dark place, performing photodynamic treatment on the fruit juice for 6 minutes by using a blue light-emitting diode array with the power of 5W, the wavelength of 435-.
After the treatment, the sample solution was serially diluted with physiological saline, 0.1mL of the diluted solution was applied to a BP medium, and the number of staphylococcus aureus was counted. The results of the experiment are shown in FIG. 2.
Example 4
Preparing 4mmol/L curcumin DMSO solution, and diluting with normal saline to obtain 40 μmol/L curcumin solution containing 10% DMSO;
diluting the prepared curcumin solution with commercially available pineapple juice to obtain a sample solution with curcumin concentration of 20 mu mol/L for later use;
selecting a staphylococcus aureus single colony in a TSB liquid culture medium, and performing shake culture at 37 ℃ for 12h to obtain a staphylococcus aureus suspension cultured to a stationary phase;
centrifuging 1ml of the bacterial solution in a centrifuge at 4 deg.C and 8000rpm for 10 min, discarding supernatant, adding equal volume of physiological saline, resuspending, and diluting with commercially available pineapple juice to obtain final bacterial concentration of 2 × 106CFU/mL；
Mixing the diluted bacteria liquid with the fruit juice obtained in the step one to ensure that the concentration of the bacteria in the final sample is 106CFU/mL, the concentration of curcumin is 10 mu mol/L;
putting 15mL of sample solution into a sterile culture dish, uniformly mixing for 30 minutes at normal temperature in a dark place, performing photodynamic treatment on the fruit juice for 12 minutes by using a blue light-emitting diode array with the power of 5W, the wavelength of 435-.
After the treatment, the sample solution was serially diluted with physiological saline, 0.1mL of the diluted solution was applied to a BP medium, and the number of staphylococcus aureus was counted. The results of the experiment are shown in FIG. 2.
As can be seen from examples 1,2,3,4 in conjunction with fig. 2, only curcumin did not exhibit any antibacterial activity against staphylococcus aureus. At 6 minutes, the curcumin-mediated photodynamic technology under the selected conditions produces good antibacterial effects on mango juice and pineapple juice, and staphylococcus aureus is reduced by 1.8 and 3.5Log CFU/mL in mango juice and pineapple juice respectively. As the illumination time increased to 12 minutes, a significant change in effect was observed, with the number of bacteria in both the mango and pineapple juices decreasing below the lowest detection limit.
In order to determine the sterilization method of the present invention, the inventors conducted a laboratory research experiment, specifically as follows:
1. sterilization effect of curcumin with different concentrations
The staphylococcus aureus strain is taken from a strain preserved in glycerol in the laboratory, and when the staphylococcus aureus strain is to be used, a ring of bacterial colony is streaked on a TSA solid culture medium and cultured in a constant temperature box at 37 ℃ for 24 hours.
A ring of well-grown single colonies was inoculated into 20mL of nutrient broth TSB liquid medium and shake-cultured at 37 ℃ for 12h to obtain an initial bacterial suspension.
Placing 6ml of bacterial liquid into a centrifuge, centrifuging at 4 deg.C and 8000rpm for 10 min, discarding supernatant, adding equal volume of physiological saline, resuspending, and continuously diluting with physiological saline to obtain final bacterial concentration of 2 × 108CFU/mL；
Preparing 8mmol/L curcumin DMSO solution, and diluting with normal saline to obtain 80 μmol/L curcumin solution containing 10% DMSO;
diluting the prepared curcumin solution into a solution with curcumin concentration of 5-40 mu mol/L by using normal saline for later use;
mixing the diluted bacterial liquid with the prepared curcumin solution to ensure that the concentration of bacteria in the final sample is 108CFU/mL, the concentration of curcumin is 2.5, 5, 7.5, 10, 20 μmol/L;
putting the sample solution into a sterile culture dish every 15mL, uniformly mixing for 30 minutes at normal temperature in a dark place, and carrying out photodynamic treatment on the fruit juice for 6 minutes by using a blue light-emitting diode array with the power of 5W, the wavelength of 435-.
After the treatment, the sample solution was serially diluted with physiological saline, 0.1mL of the diluted solution was applied to a BP medium, and the number of staphylococcus aureus was counted.
The experimental results are shown in Table 1, and 2.5 mu mol/L curcumin can generate a good sterilization effect, and the sterilization rate on staphylococcus aureus reaches 80.15%. The concentration treatments of 2.5, 5, 7.5 μmol/L gradually tended to 1 in bactericidal rate compared to the curcumin sample of 10 μmol/L concentration, but when the concentration increased above 10 μmol/L, the bactericidal rate did not change any more. Therefore, 10. mu. mol/L is the optimum concentration for sterilization under such conditions in view of practical use.
TABLE 1 Sterilization rate of Staphylococcus aureus by different curcumin concentrations
2. Sterilizing effect of different photodynamic treatment time
The staphylococcus aureus strain is taken from a strain preserved in glycerol in the laboratory, and when the staphylococcus aureus strain is to be used, a ring of bacterial colony is streaked on a TSA solid culture medium and cultured in a constant temperature box at 37 ℃ for 24 hours.
A ring of well-grown single colonies was inoculated into 20mL of nutrient broth TSB liquid medium and shake-cultured at 37 ℃ for 12h to obtain an initial bacterial suspension.
Placing 6ml of bacterial liquid into a centrifuge, centrifuging at 4 deg.C and 8000rpm for 10 min, discarding supernatant, adding equal volume of physiological saline, resuspending, and continuously diluting with physiological saline to obtain final bacterial concentration of 2 × 108CFU/mL；
Preparing 8mmol/L curcumin DMSO solution, and diluting with normal saline to obtain 80 μmol/L curcumin solution containing 10% DMSO;
diluting the prepared curcumin solution normal saline into a solution with the curcumin concentration of 20 mu mol/L for later use;
mixing the diluted bacterial liquid with the prepared curcumin solution to ensure that the concentration of bacteria in the final sample is 108CFU/mL, the concentration of curcumin is 10 mu mol/L;
putting the sample solution into a sterile culture dish every 15mL, uniformly mixing for 30 minutes at normal temperature in a dark place, using a blue light-emitting diode array, carrying out photodynamic treatment on the fruit juice for 3-15 minutes, wherein the power is 5W, the wavelength is 435-.
After the treatment, the sample solution was serially diluted with physiological saline, 0.1mL of the diluted solution was applied to a BP medium, and the number of staphylococcus aureus was counted.
As shown in Table 2, the treatment time of 6 minutes can produce good sterilization effect, and the sterilization rate of the staphylococcus aureus reaches 99.99%. As the treatment time increased, the sterilization rate increased to 1. Therefore, a treatment time of 6 minutes is the best choice for sterilization under these conditions.
TABLE 2 Sterilization Effect for different photodynamic treatment times
3. Color measurement of fruit juices under different processing conditions
Preparing 4mmol/L curcumin DMSO solution, and diluting with normal saline to obtain 40 μmol/L curcumin solution containing 10% DMSO;
diluting the prepared curcumin solution with commercially available mango juice or pineapple juice to obtain a sample solution with a final concentration of 10 mu mol/L for later use;
putting 15mL of sample solution into a sterile culture dish, uniformly mixing for 30 minutes at normal temperature in a dark place, performing photodynamic treatment on the fruit juice for 6-12 minutes by using a blue light-emitting diode array with the power of 5W, the wavelength of 435-.
After treatment, the blank was made with untreated juice. The results of the color analysis are expressed as values L a b and the results of the color determination of the photodynamic treated juice are shown in table 3 using a colorimeter.
As can be seen from the treatment of 6 minutes, the sterilization of 12 minutes has certain influence on the color of the fruit juice, so that the sterilization time is controlled to be 6 minutes, the staphylococcus aureus in the fruit juice can be effectively killed, and the product is not obviously influenced.
TABLE 3 colour change of the juice under optimum treatment conditions
Claims (5)
1. A method for cold sterilization of fruit juice by curcumin-mediated photodynamic technology is characterized by comprising the following steps:
step one, adding curcumin with different concentrations into sterile fruit juice;
step two, adding staphylococcus aureus suspension cultured to a stable stage;
and step three, after uniformly mixing the juice at normal temperature in a dark place, carrying out photodynamic treatment on the juice to obtain the sterilized juice.
2. A method of cold sterilization of fruit juice by curcumin-mediated photodynamic technology as claimed in claim 1, wherein: the first step specifically comprises:
1) preparing 4mmol/L curcumin DMSO solution, and diluting with normal saline to obtain 40 μmol/L curcumin solution containing 10% DMSO;
2) diluting the prepared curcumin solution with fruit juice to obtain sample solution with curcumin concentration of 20 μmol/L for use.
3. A method of cold sterilization of fruit juice by curcumin-mediated photodynamic technology as claimed in claim 1, wherein: the second step specifically comprises:
1) selecting a staphylococcus aureus single colony in a TSB liquid culture medium, and performing shake culture at 37 ℃ for 12h to obtain a staphylococcus aureus suspension cultured to a stable stage;
2) diluting the obtained bacterial liquid with fruit juice, and mixing with the fruit juice obtained in step one to make the final bacterial concentration 106CFU/mL, curcumin concentration of 10 u mol/L.
4. A method of cold sterilization of fruit juice by curcumin-mediated photodynamic technology as claimed in claim 1, wherein: and in the third step, uniformly mixing the juice for 30 minutes at normal temperature in a dark place, and carrying out photodynamic treatment on the juice for 6-12 minutes.
5. A method of cold sterilization of fruit juice by curcumin-mediated photodynamic technology as claimed in claim 1, wherein: the photodynamic treatment conditions are as follows: a blue LED array was used with a power of 5W, a wavelength of 435-.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115191476A (en) * | 2022-06-28 | 2022-10-18 | 江苏大学 | Dry pepper pulse strong light combined curcumin photodynamic sterilization method |
| CN115590062A (en) * | 2022-08-31 | 2023-01-13 | 广西中医药大学（Cn） | Application of emodin as photosensitizer in photodynamic sterilization |
| CN117379574A (en) * | 2023-10-13 | 2024-01-12 | 中国热带农业科学院农产品加工研究所 | Photodynamic sterilization method and application of epsilon-polylysine combined with curcumin as mediation |
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| CN115590062A (en) * | 2022-08-31 | 2023-01-13 | 广西中医药大学（Cn） | Application of emodin as photosensitizer in photodynamic sterilization |
| CN117379574A (en) * | 2023-10-13 | 2024-01-12 | 中国热带农业科学院农产品加工研究所 | Photodynamic sterilization method and application of epsilon-polylysine combined with curcumin as mediation |
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