CN115128062A - Method for detecting freshness of grains and application - Google Patents
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Abstract
The invention belongs to the technical field of chemical detection, and particularly relates to a method for detecting grain freshness and application thereof. The invention provides a method for detecting the freshness of grains aiming at the problem that the freshness of rice is difficult to quickly and accurately identify in the rice purchasing practice, which comprises the following steps: mixing luminol solution, Pbs buffer solution, grain, and H 2 O 2 After the solutions are mixed, the luminous intensity of the mixed solution is detected, and the freshness of the rice is identified according to the luminous intensity. The invention reports that a chemiluminescence analysis method is applied to detection of freshness of grains, particularly rice for the first time, and the freshness of the rice can be rapidly and accurately identified by controlling parameters such as luminol concentration, hydrogen peroxide concentration, buffer solution pH and the like.
Description
Technical Field
The invention belongs to the technical field of chemical detection, and particularly relates to a method for detecting grain freshness and application thereof.
Background
Grain safety is very important for social development. The rice is continuously subjected to various physiological changes along with the prolonging of the storage time, and the aging phenomenon of different degrees appears, thereby influencing the quality of the rice. Currently, LS/T6118 and 2017 are mainly adopted for judging the freshness of the rice, and the national standard of a rice freshness detection method does not exist. The method is only based on sensory identification and has no scientificity. The guaiacol staining method has low sensitivity for identifying paddy less than 2 years; the near infrared spectrum analysis method has expensive instrument and equipment and needs to be judged by establishing a calculation model; the pH method (bromothymol blue-methyl red) and the potassium iodide method only can roughly compare the freshness of the paddy. In the rice purchasing practice, how to identify the freshness of rice is an unprecedented challenge for the inspection and testing personnel of the basal grain depot.
Disclosure of Invention
In the rice purchasing practice, how to identify the freshness of rice is an unprecedented challenge for the inspection and testing personnel of the basal grain depot. At present, the state does not have a unified standard for detecting the freshness of the paddy, and the invention provides a method for detecting the freshness of the paddy by using a chemiluminescence analysis method aiming at the problem that the freshness of the paddy is difficult to quickly and accurately identify in the practice of purchasing the paddy, so that the quick and accurate identification of the freshness of the paddy is realized.
The technical scheme for realizing the aim of the invention is as follows:
the invention firstly provides a method for detecting the freshness of grains, which comprises the following steps:
mixing luminol solution, Pbs buffer solution, grain, and H 2 O 2 After the solutions are mixed, the luminous intensity of the mixed solution is detected, and the freshness of the grains is identified according to the luminous intensity.
Wherein the concentration of the luminol solution is 0.1 × 10 -6 ～10.0×10 -6 mol/L. Preferably, the concentration of the luminol solution is 2.0 x 10 -6 mol/L。
Wherein the pH value of the Pbs buffer solution is 6.2-11.2. Preferably, the pH of the Pbs buffer solution is 9.2.
Wherein, the grain refers to crops containing enzymes or proteins for catalyzing and enhancing a chemiluminescence system.
Preferably, the grain comprises at least one of rice, wheat, corn or soybean. Most preferably, the foodstuff is rice.
Preferably, the chemiluminescent system is a luminol-hydrogen peroxide generated chemiluminescent system.
Wherein, the H 2 O 2 The solution concentration is 0.1X 10 -3 ～10.0×10 -3 mol/L. Preferably, said H 2 O 2 The concentration of the solution is 5.0X 10 -3 mol/L。
Wherein, luminol solution, Pbs buffer solution and H 2 O 2 The volume ratio of the solution is 1-100: 2: 1 to 100. Preferably, the volume ratio is 1: 2: 1.
the grain particle number is more than or equal to 10.
Wherein, the instrument for detecting the luminous intensity of the mixed solution is a chemiluminescence analyzer for randomly detecting the luminous intensity of the solution. In a preferred embodiment of the present invention, the chemiluminescence analyzer used is an IFFM-E flow injection chemiluminescence analyzer.
The invention also provides application of the chemiluminescence analysis method in detecting the freshness of the grains.
Wherein, the grain refers to crops containing enzyme or protein for catalyzing and enhancing chemiluminescence system.
Preferably, the grain comprises at least one of rice, wheat, corn or soybean.
Has the advantages that: the invention reports that the chemiluminescence analysis method is applied to the detection of the freshness of grains, particularly rice for the first time. Meanwhile, the invention discovers that catalase can catalyze and enhance a chemiluminescence system generated by luminol-hydrogen peroxide. And it was also found that the activity of peroxidase is correlated with the freshness of rice, the longer the storage time, the lower the enzyme activity; further, the luminous intensity can be changed by controlling parameters such as the luminol concentration, the hydrogen peroxide concentration, the pH value of the buffer solution and optimizing the numerical parameter ranges of the luminol concentration, the hydrogen peroxide concentration and the buffer solution, so that the rice freshness can be furthest distinguished.
Moreover, the invention has the following advantages:
1. the detection time is extremely short, less than 1 minute, the chemiluminescence apparatus is simple, the detection is very sensitive, the dynamic range is wide, the apparatus and the equipment are cheap, and the capability of detecting the number of photons generated from the reaction is realized, which means that the extremely small amount of the analyte in the sample, such as the concentration, can be detected.
2. The excellent sensitivity and low background make the luminescence detector stand out from other analytical methods. Chemiluminescent detectors are 100,000 times more sensitive than absorption spectrometers and at least 1,000 times more sensitive than fluorometers.
3. In a luminescence reaction, there are two components of light that are able to reach the detector. The first is proportional to the finite concentration of the reactant in the chemiluminescent reaction. The second, background light, is substantially constant and is related to phosphorescence emitted by plastics, impurities, etc. in the reactants. Relative to other detection techniques such as spectrophotometers and fluorometers, the background light of chemiluminescent detectors is very low.
4. The broad kinetic range and low instrument cost are also advantages of chemiluminescent detectors. The concentration of the sample can reach more than 7 orders of magnitude in the measurement range without diluting the sample or modifying the sample cells.
5. The use is simple and convenient.
Drawings
FIG. 1 is a graph of luminescence intensities of different experimental groups in example 1 of the present invention;
FIG. 2 is a graph showing the results of detecting the freshness of rice by chemiluminescence analysis in example 3 of the present invention.
Detailed Description
Chemiluminescence analysis is a high-sensitivity trace analysis method developed in recent years, chemical energy released in a reaction is absorbed in some special chemical reactions, and light radiation is generated when a reaction intermediate or a reaction product in an electronic excited state returns to a ground state from the excited state. An analytical method for determining the content of the corresponding component in the reaction based on the luminous intensity (e.g., peak intensity) or the total amount of the luminescence at a certain time point of the chemiluminescent reaction. The luminescent substances commonly used in chemiluminescence analysis are luminol, lucigenin, lolo powder, gallic acid and oxalate peroxide. Of these, luminol is one of the most effective chemiluminescent substances recognized.
The principle is that in an alkaline environment, luminol can be oxidized by many substances to emit light, and hydrogen peroxide is the most commonly used.
The rice contains many component substances, and the difficulty in using chemiluminescence analysis for determining the freshness of rice is as follows:
1. how to determine which particular substance or substances in the rice will affect the intensity of the luminescence and further how these substances will affect the intensity of the chemiluminescence;
2. it is difficult to determine which factors, environments, or chemical components in the rice affect the freshness of the rice; and it is difficult to determine how much these factors or substances affect freshness;
3. how to relate the chemical components in the rice that affect the intensity of the luminescent substances to the chemical substances, environments, or factors in the rice that affect the freshness of the rice.
In order to solve the difficulties, through research and a large number of experimental screens, the applicant finds that peroxidase in rice grains can catalyze and enhance a chemiluminescence system generated by luminol-hydrogen peroxide. It was also found that the peroxidase activity is correlated with the freshness of rice, and the longer the storage time, the lower the enzyme activity, and therefore, the freshness of rice can be judged from the chemiluminescence intensity. The rice freshness is determined by utilizing a luminol chemiluminescence system, and the rice freshness is identified by detecting the rice containing different peroxidase activities.
Meanwhile, the applicant consults the prior art to find that the freshness of the rice is determined by not peroxidase but lipase, because fatty acid is the most important index for judging the freshness of the rice, and the higher the fatty acid value is, the fresher the rice is. Since the fatty acid is produced by increasing the content of fatty acid under the catalysis of lipase, which causes rice to rot, the enzyme which is determined to be the decisive enzyme in the current method for detecting the freshness of rice should be lipase.
In addition, the applicant also finds that not only the activity of the enzyme can determine the intensity of chemiluminescence, but also the change of luminol concentration, hydrogen peroxide concentration and buffer solution pH can cause the change of luminous intensity, the differentiation of rice freshness can be realized in a proper range, and the quick and accurate identification of the rice freshness can be realized by optimizing the experimental conditions influencing the luminous intensity of the system. The applicant finds that the following luminol concentration, hydrogen peroxide concentration and buffer solution pH can better identify the freshness of the rice through a large number of screening experiments:
the concentration of luminol solution is 0.1 × 10 -6 ～10.0×10 -6 mol/L. Preferably, the concentration of the luminol solution is 2.0 × 10 -6 mol/L。
The pH value of the Pbs buffer solution is 6.2-11.2. Preferably, the pH of the Pbs buffer solution is 9.2.
H 2 O 2 The concentration of the solution is 0.1X 10 -3 ～10.0×10 -3 mol/L. Preferably, H 2 O 2 The concentration of the solution is 5.0X 10 -3 mol/L。
Wherein, the Pbs buffer solution comprises the following chemical components: na (Na) 2 HPO 4 、KH 2 PO 4 NaCl and KCl (purchased from Shanghai Co., Ltd., Biotechnology).
The amount (i.e., amount) of the rice grains has some effect on the chemiluminescence intensity, but is not critical, because applicants have found that when the number of rice grains is small, the chemiluminescence intensity gradually increases as the number of grains increases; when the number of the rice grains is more than 10, the chemiluminescence intensity changes less along with the increase of the number of the grains. That is, the amount of the rice grains (i.e., the enzyme content) in the same sample is not a determining factor mainly affecting the chemiluminescence intensity, but the storage time, i.e., the enzyme activity, is greatly affected, so that the number of the rice grains taken is 10.
The invention uses luminol solution, Pbs buffer solution, grains, especially rice grains, and H 2 O 2 After the solutions are mixed, the luminous intensity of the mixed solution is detected by a chemiluminescence instrument, and compared with the prior art, the method for identifying the freshness of the rice according to the luminous intensity has the advantages that:
1. the detection time is very short (less than 1 minute), the chemiluminescence instrument is simple, the detection is very sensitive (a photomultiplier is arranged as a detector, and the detection sensitivity of the photomultiplier can reach 10) -9 Above the order of mol/L), a wide kinetic range, inexpensive instrumentation, the ability to detect the number of photons generated from the reaction, means that extremely small aliquots (concentrations) of analyte in the sample can be detected.
2. The excellent sensitivity and low background make the luminescence detector stand out from other analytical methods. Chemiluminescent detectors are 100,000 times more sensitive than absorption spectrometers and at least 1,000 times more sensitive than fluorometers.
3. In a luminescence reaction, there are two components of light that are able to reach the detector. The first is proportional to the finite concentration of the reactant in the chemiluminescent reaction. The second, background light, is essentially invariant and is related to phosphorescence from plastics, impurities, etc. in the reactants. Relative to other detection techniques such as spectrophotometers and fluorometers, the background light of chemiluminescent detectors is very low.
4. The broad kinetic range and low instrument cost are also advantages of chemiluminescent detectors. The concentration of the sample can reach more than 7 orders of magnitude in the measurement range without diluting the sample or modifying the sample cells.
5. The use is simple and convenient.
The key point of the invention is that peroxidase in the rice catalyzes and enhances a chemiluminescence system generated by luminol-hydrogen peroxide, so that the enhancement of the chemiluminescence reaction system (namely, the enhancement of a chemiluminescence signal generated in an original reaction system) caused by the catalytic action is replaced by related enzymes in grains (including rice, wheat, corn, soybean and the like) or proteins which have the same action with the enzymes. It should be particularly noted that the chemiluminescent reaction system is not limited to the luminol-hydrogen peroxide system, and that other luminescent system enhancements that function as equivalent reagents are also contemplated.
Methods similar to the method of the invention and the conversion of the results to instruments involved by the method are also within the scope of the invention.
The scheme of the invention will be explained with reference to the examples. It will be appreciated by those skilled in the art that the following examples are illustrative of the invention only and should not be taken as limiting the scope of the invention. The examples, where specific techniques or conditions are not indicated, are to be construed according to the techniques or conditions described in the literature in the art or according to the product specifications. The reagents or instruments used are conventional products which are commercially available, and are not indicated by manufacturers.
Example 1 demonstrates that rice grain particles can enhance luminol chemiluminescence systems
Respectively prepared at a concentration of 1.0 × 10 -3 mol/L H 2 O 2 Solution, 2.0X 10 -6 mol/L luminol solution andand Pbs buffer solution with the pH value of 8.2 are used for verification, so that peroxidase in the rice grains can catalyze the luminol hydrogen peroxide system. The specific operation is as follows:
1. 1.0 mL of 2.0X 10 was taken -6 mol/L luminol solution, 2.0 mL of Pbs buffer solution having a pH of 8.2 was added, and then rice grains (10 grains) which were hulled and cut to have an exposed cross section were added, and the luminous intensity thereof was measured by a chemiluminescence apparatus (IFFM-E type flow injection chemiluminescence analyzer), and a signal value was found to be 0.
2. 2.0 mL of Pbs buffer solution with pH of 8.2 was taken, rice grains (10 grains) with an exposed cross section were added, and 1.0 mL of 1.0X 10 was introduced -3 mol/L H 2 O 2 The solution, the luminescence intensity of which was measured by a chemiluminescence apparatus, was found to have a signal value of almost 0.
3. 1.0 mL of 2.0X 10 was taken -6 mol/L luminol solution, 2.0 mL Pbs buffer solution with pH of 8.2 was added, and 1.0 mL 1.0X 10 was added -3 mol/L H 2 O 2 The solution was measured for luminescence intensity by a chemiluminescence apparatus, and found to have a signal value of 377.3.
4. 1.0 mL of 2.0X 10 was taken -6 mol/L luminol solution, 2.0 mL of Pbs buffer solution with pH 8.2 was added, then rice grains (10 grains) with exposed cross-section were added, and finally 1.0 mL of 1.0X 10 was passed -3 mol/L H 2 O 2 The solution was measured for luminescence intensity by a chemiluminescence apparatus, and found to have a signal value of 3322.7. The specific data are shown in fig. 1.
Example 2 study of luminol chemiluminescence assay conditions
1、H 2 O 2 Influence of solution concentration on luminous intensity of system
1.0 mL of 2.0X 10 was taken -6 mol/L luminol solution, 2.0 mL Pbs buffer solution with pH of 8.2 was added, then rice grains (10 grains) with exposed cross-section were added, and finally 1.0 mL 0.1X 10 grains were separately introduced -3 mol/L H 2 O 2 、0.2 ×10 -3 mol/L H 2 O 2 、0.5 ×10 -3 mol/L H 2 O 2 、1.0 ×10 -3 mol/L H 2 O 2 、2.0 ×10 -3 mol/L H 2 O 2 、5.0 ×10 -3 mol/L H 2 O 2 、10.0 ×10 -3 mol/L H 2 O 2 The solution, the intensity of the luminescence measured by a chemiluminescence apparatus, the value of the luminescence signal is shown in Table 1, as H 2 O 2 The solution concentration increases, the luminous intensity increases, at H 2 O 2 The concentration of the solution is 5.0X 10 -3 At mol/L, the signal value reaches a maximum, and H 2 O 2 The concentration of the solution is 5.0X 10 -3 At mol/L and above, there is little increase in signal value, and therefore, the optimum H for the experiment is finally selected 2 O 2 The concentration of the solution is 5.0X 10 -3 mol/L。
TABLE 1H 2 O 2 Influence of solution concentration on luminous intensity of system
| H 2 O 2 Concentration of solution (10) -3 mol/L） | Luminous intensity (a.u.) |
| 0.1 | 135.67 |
| 0.2 | 392.33 |
| 0.5 | 854.00 |
| 1.0 | 2691.67 |
| 2.0 | 4325.00 |
| 5.0 | 18584.00 |
| 10.0 | 18632.00 |
2. Influence of luminol solution concentration on system luminous intensity
Respectively take 1.0 mL of 0.1X 10 -6 mol/L、0.2 ×10 -6 mol/L、0.5 ×10 -6 mol/L、1.0 ×10 -6 mol/L、2.0 ×10 -6 mol/L、5.0 ×10 -6 mol/L、10.0 ×10 -6 mol/L luminol solution, 2.0 mL of Pbs buffer solution with pH 8.2 was added, then rice grains (10 grains) with exposed cross-section were added, and finally 1.0 mL of 5.0X 10 was added -3 mol/L H 2 O 2 The solution is detected by a chemiluminescence instrument to obtain the luminous intensity, the luminous signal value is shown in table 2, the luminous signal value is enhanced along with the increase of the concentration of the luminol solution, and the concentration of the luminol solution is 2.0 multiplied by 10 -6 At mol/L, the maximum is reached, and then the signal value is basically kept unchanged along with the increase of the concentration of the luminol solution. Therefore, the concentration of the luminol solution that is the best for the final selection experiment is 2.0 × 10 -6 mol/L。
TABLE 2 influence of luminol solution concentration on the luminous intensity of the system
| Luminol solution concentration (10) -6 mol/L） | Luminous intensity (a.u.) |
| 0.1 | 973.67 |
| 0.2 | 2521.33 |
| 0.5 | 4717.33 |
| 1.0 | 8846.00 |
| 2.0 | 17938.33 |
| 5.0 | 18692.33 |
| 10.0 | 18471.00 |
3. Influence of Pbs buffer solution pH value on system luminous intensity
1.0 mL of 2.0X 10 was taken -6 Adding 2.0 mL of Pbs buffer solution with pH of 6.2, 7.2, 8.2, 9.2, 10.2 to mol/L of luminol solution, respectively, adding rice grains (10 grains) with exposed cross section, and introducing 1.0 mL of 5.0 × 10 -3 mol/L H 2 O 2 The solution is detected by a chemiluminescence instrument to obtain the luminous intensity, the luminous signal value is shown in table 3, the more alkaline the pH value is, the stronger the luminous intensity of the system is, when the pH is 9.2, the luminous intensity is larger, and then the luminous intensity is slowly increased along with the increase of the pH, but the enhancement rate is very small and basically negligible. Therefore, when the pH value of the Pbs buffer solution is 9.2, the luminous intensity of the whole system is optimal. Therefore, the pH of the Pbs buffer solution that was the best for the experiment was finally selected to be 9.2.
TABLE 3 influence of pH value of Pbs buffer solution on luminous intensity of system
| pH value of Pbs buffer solution | Luminous intensity (a.u.) |
| 6.2 | 106.00 |
| 7.2 | 2406.33 |
| 8.2 | 18679.33 |
| 9.2 | 24891.67 |
| 10.2 | 25999.00 |
4. Influence of the amount of Rice particles on the luminous intensity of the System
1.0 mL of 2.0X 10 was taken -6 Adding 2.0 mL of Pbs buffer solution with pH of 9.2 to mol/L of luminol solution, adding 2.5, 5, 7.5, 10, 12.5, 15, and 17.5 rice grains with exposed cross-section, respectively, and introducing 1.0 mL of 5.0 × 10 -3 mol/L H 2 O 2 The solution was measured for luminescence intensity by a chemiluminescence apparatus, and the luminescence signal values are shown in Table 4. When the number of the rice particles is less, the chemiluminescence intensity gradually increases along with the increase of the number of the particles; when the number of the rice grains is more than 10, the chemiluminescence intensity changes less along with the increase of the number of the grains. It can be known from this that when 10 grains of rice grains are addedThe luminous intensity of the whole system reaches the best; it was also shown that the amount of rice grains (i.e., the enzyme content) in the same sample was not a determining factor that mainly affected the intensity of chemiluminescence, but the storage time, i.e., the enzyme activity, was greatly affected. Therefore, the number of the rice grains which is the best for the experiment is finally selected to be 10 grains.
TABLE 4 influence of the number of rice grains on the luminous intensity of the system
| Number of rice grains (grain) | Luminous intensity (a.u.) |
| 2.5 | 11026.33 |
| 5.0 | 17344.67 |
| 7.5 | 19146.00 |
| 10.0 | 25168.67 |
| 12.5 | 24182.33 |
| 15.0 | 24890.67 |
| 17.5 | 24529.33 |
Example 3 determination of Rice freshness Using luminol chemiluminescence System
In this example 3, the freshness of rice is judged according to the chemiluminescence intensity; and the method is verified to realize the differentiation of rice samples with different freshness.
1. 1.0 mL of 2.0X 10 was taken -6 Adding 2.0 mL of Pbs buffer solution with pH of 9.2 into mol/L luminol solution, respectively, adding husked indica rice grains (10 grains, with age of 2019 and 2021, origin of Jiangxi), cutting to expose cross section, and introducing 1.0 mL of 5.0 × 10 -3 mol/L H 2 O 2 The solution is detected by a chemiluminescence instrument to obtain signal values of 13636 and 19073, wherein the high luminescence intensity value is indica rice in 2021 years, and the low luminescence intensity value is indica rice in 2019 years, so that the solution can be distinguished by the method.
2. 1.0 mL of 2.0X 10 was taken -6 Adding 2.0 mL of Pbs buffer solution with pH of 9.2 into mol/L luminol solution, respectively, adding hulled and cut semen oryzae Sativae granules (10 granules, with age of 2017 and 2021 years, origin of Heilongjiang), introducing 1.0 mL of 5.0 × 10 -3 mol/L H 2 O 2 The solution is detected by a chemiluminescence instrument to obtain signal values of 15921.3 and 23130.7, wherein the signal values are respectively 891 japonica rice with high luminescence intensity and 2017 japonica rice with low luminescence intensity, and therefore, the solution can be distinguished by the method.
3. 1.0 mL of 2.0X 10 was taken -6 Adding 2.0 mL of Pbs buffer solution with pH of 9.2 into mol/L luminol solution, respectively, adding hulled indica rice grains (10 grains, with age of 2017 and 2021 year, origin of Hubei) with exposed cross section, and introducing 1.0 mL of 5.0 × 10 -3 mol/L H 2 O 2 The solution is detected by a chemiluminescence instrument to obtain signal values of 10944.7 and 15156.7, wherein the signal values are indica rice in 2021 years with high luminescence intensity and indica rice in 2017 years with low luminescence intensity, and therefore, the solution can be distinguished by the method. The specific data distribution is shown in fig. 2.
According to the results of the method, the method can distinguish the rice samples with different freshness.
The above is only a preferred embodiment of the present invention, and it should be noted that the above preferred embodiment should not be considered as limiting the present invention, the scope of the present invention should be defined by the scope defined by the claims, and it should be apparent to those skilled in the art that several modifications and embellishments can be made without departing from the spirit and scope of the present invention, and these modifications and embellishments should also be considered as the scope of the present invention.
Claims (9)
1. The method for detecting the freshness of the grains is characterized by comprising the following steps: the method comprises the following steps:
mixing luminol solution, Pbs buffer solution, grain, and H 2 O 2 After the solutions are mixed, the luminous intensity of the mixed solution is detected, and the freshness of the grains is identified according to the luminous intensity.
2. The method for detecting food freshness according to claim 1, wherein: the concentration of the luminol solution is 0.1 multiplied by 10 -6 ～10.0×10 -6 mol/L; preferably, the concentration of the luminol solution is 2.0 x 10 -6 mol/L。
3. The method for detecting food freshness according to claim 1 or 2, wherein: the pH value of the Pbs buffer solution is 6.2-11.2; preferably, the pH of the Pbs buffer solution is 9.2.
4. The method for detecting food freshness according to claim 1, wherein: the grain refers to crops containing enzymes or proteins for catalyzing and enhancing a chemiluminescence system; preferably, the grain comprises at least one of rice, wheat, corn or soybean; most preferably, the foodstuff is rice.
5. The method for detecting food freshness according to claim 4, wherein: the chemical luminous system is a luminol-hydrogen peroxide generated chemical luminous system.
6. The method for detecting food freshness according to claim 1, wherein: said H 2 O 2 The concentration of the solution is 0.1X 10 -3 ～10.0×10 -3 mol/L; preferably, said H 2 O 2 The solution concentration was 5.0X 10 -3 mol/L。
7. The method for detecting grain freshness according to claim 1, wherein: luminol solution, Pbs buffer solution, H 2 O 2 The volume ratio of the solution is 1-100: 2: 1-100; preferably, the volume ratio is 1: 2: 1.
8. the method for detecting food freshness according to claim 1, wherein: the grain particle number is more than or equal to 10.
9. Use of the method for detecting food freshness according to any one of claims 1 to 8 in food freshness detection.
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