Ecotoxicity monitoring device using luminescent bacteria and ecotoxicity monitoring method using the same

Abstract

To provide an ecotoxicity monitoring device using a luminescent bacterium and an ecotoxicity monitoring method using the same.SOLUTION: An ecotoxicity monitoring device using a luminescent bacterium and an ecotoxicity monitoring method using the same are provided. The ecotoxicity monitoring device comprises: a reaction tank module including a reference reaction tank, a sample reaction tank, and a dilution reaction tank, and supplies a mixed solution containing the luminescent bacterium; a luminescent bacterium culture portion; a diluted water supply portion; a sample supply portion; an optical measurement portion that measures the amount of luminescence of the luminescent bacterium in a toxicity assessment substance provided from the sample reaction tank and the dilution reaction tank; a control portion; and an analysis portion that determines whether a toxic substance is present a sample on the basis of a light quantity value provided from the optical measurement portion. After time predetermined as an initial light quantity value of the reference reaction tank measured through the optical measurement portion has elapsed, the analysis portion is configured to calculate a correction coefficient through the measured final light quantity value, and calculate a corrected light quantity value of the sample reaction tank and dilution reaction tank through the calculated correction coefficient.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to an ecotoxicity monitoring device using luminescent bacteria and an ecotoxicity monitoring method using the same, and more particularly to an ecotoxicity monitoring device using luminescent bacteria that is adapted to quickly and accurately monitor the presence or absence of toxicity in a sample and an ecotoxicity monitoring method using the same. [Background technology]

[0002] Although sewage and wastewater treatment facilities have been established and operated to improve water quality and protect the ecosystem, there is a problem in that harmful chemicals cannot be completely removed through biological or physicochemical treatment processes. Therefore, harmful chemicals that are not removed during the sewage and wastewater treatment process are discharged into public water bodies, causing adverse effects on the ecosystem and public health.

[0003] In addition, hazardous / noxious substances (HNS) released into the ocean due to marine industrial facilities or ship accidents cause direct and indirect damage to the surrounding ocean and seabed, marine users, and coastal residents. Such hazardous / noxious substances released into the ocean are diverse, and physicochemical analysis alone without prior information provides limited information on the types and properties of substances. Furthermore, physicochemical analysis techniques are practically impossible to analyze for toxic substances in all samples due to the cost, time, technology, and manpower required for analysis.

[0004] Therefore, ecotoxicity assessment and analysis techniques are emerging as a scientific and efficient management method for environmental analysis. These ecotoxicity assessment and analysis techniques use living organisms to evaluate the hazards of substances present in the environment.

[0005] However, conventional ecotoxicity assessment devices have a problem in that they take at least 15 minutes to assess whether or not a toxic substance is present in a sample, even if it is the fastest. As such, the longer the assessment time is when assessing whether or not a toxic substance is present in a sample using an ecotoxicity assessment device, the more difficult it is to respond quickly to environmental pollution and the greater the damage that may result.

[0006] Therefore, various research and development efforts are being conducted on ecotoxicity monitoring devices that can monitor hazardous / harmful substances that are released into not only rivers but also the ocean more quickly and accurately. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] Korean Patent Publication No. 2014-0093389 (2014.07.28) Summary of the Invention [Problem to be solved by the invention]

[0008] The technical objective of the present invention to solve the above problems is to provide an ecotoxicity monitoring device using luminescent bacteria that can quickly and accurately monitor the presence or absence of toxicity in a sample, and an ecotoxicity monitoring method using the same. [Means for solving the problem]

[0009] In order to achieve the above technical objectives, one embodiment of the present invention provides a reaction vessel module including a reference reaction vessel, a sample reaction vessel, and a dilution reaction vessel, and a bacterial dilution vessel for supplying a mixture containing luminescent bacteria to the reference reaction vessel, the sample reaction vessel, and the dilution reaction vessel; a luminescent bacteria culturing unit for supplying luminescent bacteria for detecting the toxicity of a sample to the bacterial dilution vessel; a dilution water supply unit for supplying dilution water to the bacterial dilution vessel, the reference reaction vessel, and the dilution reaction vessel; a sample supply unit for supplying a sample to the sample reaction vessel and the dilution reaction vessel; and an analysis unit that determines whether or not a toxic substance is present in a sample based on the light intensity value provided by the optical measurement unit, wherein the analysis unit calculates a correction coefficient based on the initial light intensity value of the reference reaction tank measured by the optical measurement unit and the final light intensity value measured after a predetermined time has elapsed, and calculates corrected light intensity values ​​of the sample reaction tank and the dilution reaction tank based on the calculated correction coefficient.

[0010] In one embodiment of the present invention, the analysis unit calculates the toxicity index (TI s ) is calculated, but the toxicity index (TI s ) is calculated by calculating the correction coefficient of the reference reaction vessel according to the following formula 1, calculating the corrected light intensity value of the sample reaction vessel to which the correction coefficient is applied according to the following formula 2, and calculating the toxicity index (TI) according to the following formula 3 using the final light intensity value of the sample reaction vessel measured after a predetermined time has elapsed since the required sample was injected into the sample reaction vessel from the sample supply unit. s ) can be calculated.

[0011]

number

[0012]

number

[0013] [S0 = initial light intensity value after 1 minute of the mixture supplied from the bacterial dilution chamber to the sample reaction chamber, C0 = correction coefficient]

[0014]

number

[0015] [S1 = corrected light intensity value after the first minute of the sample reaction chamber to which the correction coefficient has been applied, S2 = final light intensity value after a predetermined time has elapsed with a fixed amount of sample injected into the mixed solution in the sample reaction chamber]

[0016] In one embodiment of the present invention, the analysis unit calculates the toxicity index (TI d ) is calculated, but the toxicity index (TI d ) is calculated by calculating the corrected light intensity value of the dilution reaction vessel to which the correction coefficient of the reference reaction vessel is applied according to the following formula 4, and then calculating the toxicity index (TI) from the final light intensity value of the dilution reaction vessel measured after a predetermined time has elapsed after the sample and dilution water are injected into the dilution reaction vessel at a required ratio according to the following formula 5. d ) can be calculated.

[0017]

number

[0018] [D0 = initial light intensity value after 1 minute of the mixed solution supplied from the bacterial dilution tank to the dilution reaction tank, C0 = correction coefficient]

[0019]

number

[0020] [D1 = corrected light intensity value after the first minute of the dilution reaction vessel to which the correction coefficient has been applied, D2 = final light intensity value after a predetermined time has elapsed with the sample and dilution water injected into the mixture in the dilution reaction vessel]

[0021] In one embodiment of the present invention, the analysis unit determines that the calculated correction coefficient is valid when it is 0.6 to 1.3, and calculates the toxicity index (TI s ) is between 20 and 80%, it can be determined that toxic substances are present in the sample.

[0022] In one embodiment of the present invention, the analysis unit calculates the toxicity index (TI s ) and the toxicity index (TI d ) the toxicity risk of the sample can be assessed through comparison.

[0023] In one embodiment of the present invention, the predetermined time for measuring the final light intensity values ​​of the reference reaction chamber, the sample reaction chamber and the dilution reaction chamber may be 5 minutes.

[0024] One embodiment of the present invention includes the steps of: (A) preparing a mixture of luminescent bacteria and dilution water in a bacterial dilution tank; (B) supplying the mixture from the bacterial dilution tank to a reference reaction tank, a sample reaction tank, and a dilution reaction tank, respectively; (C) measuring initial light intensity values ​​of the luminescent bacteria in the reference reaction tank, the sample reaction tank, and the dilution reaction tank through an optical measurement unit after a predetermined time has elapsed; (D) after measuring the initial light intensity values, supplying dilution water to the reference reaction tank, supplying a sample to the sample reaction tank, and supplying a predetermined amount of sample and dilution water to the dilution reaction tank; and (E) measuring the initial light intensity values ​​through the optical measurement unit after a predetermined time has elapsed. and (F) providing the initial and final light intensity values ​​of the luminescent bacteria in the reference reaction tank, the sample reaction tank, and the dilution reaction tank with the optical measurement unit, and determining whether or not there is toxicity in the sample, wherein the analysis unit calculates a correction coefficient based on the initial and final light intensity values ​​of the reference reaction tank measured by the optical measurement unit, calculates corrected light intensity values ​​for the sample reaction tank and the dilution reaction tank using the calculated correction coefficient, and determines a toxicity index for the toxicity evaluation substance in the sample reaction tank and the dilution reaction tank. [Effects of the Invention]

[0025] The effects of the ecotoxicity monitoring device using luminescent bacteria and the ecotoxicity monitoring method using the same according to the present invention, which are capable of quickly and accurately monitoring the presence or absence of toxicity in a sample, will be described as follows.

[0026] According to the present invention, the analysis unit calculates a correction coefficient based on the initial and final light intensity values ​​measured from the reference reaction tank, and can quickly calculate the toxicity index for the toxicity evaluation substance in the sample reaction tank and the dilution reaction tank using the calculated correction coefficient. Such an ecotoxicity monitoring device using luminescent bacteria can evaluate the presence or absence of a toxic substance in a sample within five minutes, allowing for quick response to dangerous / harmful substances that may be discharged into rivers or the ocean.

[0027] The effects of the present invention are not limited to the effects described above, but must be understood to include all effects that can be inferred from the configuration of the invention described in the detailed description of the present invention or the claims. [Brief explanation of the drawings]

[0028] [Figure 1] 1 is a block diagram of an ecotoxicity monitoring device using luminescent bacteria according to an embodiment of the present invention; [Figure 2] 1 is an exemplary diagram of an ecotoxicity monitoring device using luminescent bacteria according to an embodiment of the present invention; [Figure 3] FIG. 10 is an exemplary diagram illustrating a process in which an initial light intensity value is measured after a mixture contained in a bacterial dilution tank according to one embodiment of the present invention is supplied to a reference reaction tank, a sample reaction tank, and a dilution reaction tank. [Figure 4] 10 is an exemplary diagram illustrating a process in which the final light intensity value is measured after the required sample and dilution water are supplied to the reference reaction chamber, sample reaction chamber, and dilution reaction chamber according to one embodiment of the present invention. FIG. [Figure 5] 1 is a flowchart illustrating a method for monitoring ecotoxicity using luminescent bacteria according to an embodiment of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0029] The present invention will now be described with reference to the accompanying drawings. However, the present invention may be embodied in various different forms and is not limited to the embodiments described herein. In addition, in order to clearly illustrate the present invention in the drawings, parts that are not relevant to the description are omitted, and similar parts are designated by similar reference numerals throughout the specification.

[0030] Throughout the specification, when a part is said to be "connected" to another part, this includes not only when it is "directly connected" to another part, but also when it is "indirectly connected" via another member in between. Furthermore, when a part is said to "comprise" a certain element, this does not mean that it excludes other elements, but that it may further comprise other elements, unless otherwise specified.

[0031] In the present invention, the terms "upper" and "lower" refer to positions above or below the target member, and do not necessarily refer to positions above or below the direction of gravity.

[0032] Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings.

[0033] FIG. 1 is a configuration diagram of an ecotoxicity monitoring device using luminescent bacteria according to one embodiment of the present invention; FIG. 2 is an exemplary diagram of an ecotoxicity monitoring device using luminescent bacteria according to one embodiment of the present invention; FIG. 3 is an exemplary diagram showing a process in which an initial light intensity value is measured after a mixed solution contained in a bacterial dilution tank according to one embodiment of the present invention is supplied to a reference reaction tank, a sample reaction tank, and a dilution reaction tank; and FIG. 4 is an exemplary diagram showing a process in which a final light intensity value is measured after the required sample and dilution water are supplied to a reference reaction tank, a sample reaction tank, and a dilution reaction tank according to one embodiment of the present invention.

[0034] As shown in Figures 1 to 4, an ecotoxicity monitoring device 1000 using luminescent bacteria may include a reaction tank module 100, a luminescent bacteria cultivation section 200, a dilution water supply section 300, a sample supply section 400, an optical measurement section 500, a control section 600, and an analysis section 700.

[0035] The ecotoxicity monitoring device 1000 may be provided with a casing 810 for protecting various internal equipment from the outside.

[0036] The inside of such casing 810 may be partitioned, and for example, a monitor unit 820 that displays toxicity measurement results may be provided on the upper side, an analysis unit 700 may be provided below the monitor unit 820, an autosampler A may be provided below the analysis unit 700, and a reagent and waste liquid storage unit 830 may be stored below the autosampler A. The arrangement of each component provided within such casing 810 does not necessarily have to be configured in the form mentioned above, and may be configured in various other forms.

[0037] Here, the autosampler A equipped with the reaction vessel module 100 and the optical measurement unit 500 provided in the ecotoxicity monitoring device 1000 can be easily pulled out to the outside, for example, by sliding, or pulled into the casing 810 from the outside. With the autosampler A being able to slide in this way, if an abnormality occurs in any of the multiple components provided in the autosampler A, the user can easily repair or replace the malfunctioning component while the autosampler A itself is pulled out to the outside.

[0038] The ecotoxicity monitoring device 1000 uses luminescent bacteria to determine whether or not a toxic substance is present in a sample. That is, the ecotoxicity monitoring device 1000 measures the rate of change in the luminescence intensity of the luminescent bacteria when a sample is added to determine whether or not a toxic substance is present in the sample. While determining whether or not a toxic substance is present in a sample based on the rate of change in the luminescence intensity of the luminescent bacteria has been disclosed in the prior art, the analysis unit 700 included in the ecotoxicity monitoring device 1000 of the present invention calculates a correction coefficient based on the initial and final light intensity values ​​measured in the reference reaction chamber 120, and more quickly determines whether or not a toxic substance is present in the toxicity evaluation substances in the sample reaction chamber 130 and the dilution reaction chamber 140 using the calculated correction coefficient.

[0039] Specifically, the reaction chamber module 100 may include a bacterial dilution chamber 110 , a reference reaction chamber 120 , a sample reaction chamber 130 and a dilution reaction chamber 140 .

[0040] Here, the bacteria dilution tank 110 is provided with a predetermined storage space, and a mixture containing luminescent bacteria and dilution water is stored inside the bacteria dilution tank 110. The luminescent bacteria are supplied to the bacteria dilution tank 110 from the luminescent bacteria culturing unit 200, and dilution water is supplied from the dilution water supply unit 300.

[0041] Here, luminescent bacteria that detect the toxicity of a sample are cultivated in the luminescent bacteria culturing unit 200. The luminescent bacteria culturing unit 200 is maintained at, for example, about 4° C. to inhibit growth.

[0042] The diluting water supply unit 300 is configured to store diluting water, such as mineral water. The diluting water stored in the diluting water supply unit 300 does not necessarily have to consist of mineral water alone, and may store salt water (NaCl) or the like.

[0043] The mixture containing the luminescent bacteria and dilution water supplied to the bacteria dilution tank 110 may be mixed uniformly through an agitator provided in the bacteria dilution tank 110 .

[0044] The bacterial dilution tank 110 is configured to send a mixture of luminescent bacteria and dilution water in a predetermined ratio to the reference reaction tank 120, the sample reaction tank 130, and the dilution reaction tank 140, respectively. The mixture sent from the bacterial dilution tank 110 to each of the reference reaction tank 120, the sample reaction tank 130, and the dilution reaction tank 140 may be sent, for example, through a transfer pump (not shown). The mixture, as well as the sample and / or dilution water described below, may be moved by operation of the respective transfer pumps connected to the sample supply unit 400 and / or the dilution water supply unit 300, etc.

[0045] In addition, after a predetermined time (1 minute) has elapsed, the mixed liquid sent to the reference reaction chamber 120, sample reaction chamber 130, and dilution reaction chamber 140 is measured for the initial light intensity value of the luminescent bacteria in each reaction chamber through the optical measurement unit 500.

[0046] The optical measuring unit 500 includes an optical sensor unit 510 that measures the amount of minute luminescence from the luminescent bacteria that has flowed into the internal cell, and provides the measured initial light amount value to the analyzing unit 700.

[0047] Here, the cell may be made of quartz material to optimally detect the amount of light. The cell may also be formed in a darkroom to detect emitted light. The optical sensor unit 510 may be, for example, a PMT (Photo Multiplier Tube). The optical sensor unit 510 measures the initial amount of luminescence of the luminescent bacteria when no sample is added and the final amount of luminescence of the luminescent bacteria after a predetermined time has elapsed when a sample is added.

[0048] The number of such optical sensor units 510 may correspond to the number of the plurality of reaction chambers, i.e., the number of optical sensor units 510 may correspond to the number of reference reaction chambers 120, sample reaction chambers 130, and dilution reaction chambers 140.

[0049] Here, the reference reaction chamber 120, the sample reaction chamber 130, and the dilution reaction chamber 140 are in communication with the respective optical sensor units 510. Thus, a mixture of the luminescent bacteria contained in the reference reaction chamber 120, the sample reaction chamber 130, and the dilution reaction chamber 140 may be supplied to the respective optical sensor units 510. Since each of the optical sensor units 510 is in communication with the reference reaction chamber 120, the sample reaction chamber 130, and the dilution reaction chamber 140, the mixture contained in the cell of the optical sensor unit 510 after light intensity measurement has been completed may be discharged to the respective reaction chambers.

[0050] The light intensity value of each reaction vessel measured by the optical sensor unit 510 in this manner is provided to the analysis unit 700.

[0051] After the light intensity measurement for the mixed solution is completed, the cell in the light sensor unit 510 may be diluted with dilution water provided from the dilution water supply unit 300. That is, the light sensor unit 510 is configured to be cleaned before a new light intensity measurement. Thus, the light sensor unit 510 can accurately measure the light intensity value for the provided evaluation substance without being affected by previously used samples or luminescent bacteria.

[0052] After the measurement of the initial light intensity values ​​for the mixed solutions provided from the reference reaction chamber 120, the sample reaction chamber 130, and the dilution reaction chamber 140 has been completed, a predetermined amount of dilution water is further supplied to the reference reaction chamber 120, a sample is supplied to the sample reaction chamber 130, and the sample and dilution water are supplied to the dilution reaction chamber 140 at a constant ratio. Here, the sample supply unit 400 supplies the sample only to the sample reaction chamber 130 and the dilution reaction chamber 140. The evaluation substance in the sample reaction chamber 130 and the dilution reaction chamber 140 to which the sample has been further supplied may be a toxicity evaluation substance.

[0053] In this manner, after a certain period of time (5 minutes) has elapsed while further supplying predetermined amounts of sample and / or dilution water to the reference reaction chamber 120, sample reaction chamber 130, and dilution reaction chamber 140, the final light intensity values ​​are measured for the evaluation substance in the reference reaction chamber 120 to which only dilution water has been added, and for the toxicity evaluation substance in the sample reaction chamber 130 and dilution reaction chamber 140.

[0054] In this way, each final light intensity value measured by the optical sensor unit 510 is provided to the analysis unit 700. In addition, the evaluation substance / toxicity evaluation substance contained in the optical sensor unit 510 is discharged into the corresponding reaction vessel, and the cell in the optical sensor unit 510 is washed with dilution water.

[0055] After the final light intensity value for each reaction chamber is measured, the evaluation substances / toxicity evaluation substances contained in the reference reaction chamber 120, sample reaction chamber 130, and dilution reaction chamber 140 are discharged to the waste liquid storage unit 830 via a transfer pump. In addition, the reference reaction chamber 120, sample reaction chamber 130, and dilution reaction chamber 140 are washed with dilution water in preparation for the next evaluation test.

[0056] Meanwhile, the control unit 600 controls the operation of various components provided in the ecotoxicity monitoring device 1000, such as the reaction vessel module 100, the luminescent bacteria culturing unit 200, the dilution water supply unit 300, the sample supply unit 400, and the optical measurement unit 500. The control unit 600 controls various operations, such as supplying sample and / or dilution water to the reaction vessel module 100 or cleaning the optical sensor unit 510.

[0057] Meanwhile, the analysis unit 700 calculates a toxicity index based on the initial light intensity values ​​and final light intensity values ​​measured from the evaluation substance / toxicity evaluation substance provided to the optical sensor unit 510 from the reference reaction chamber 120, sample reaction chamber 130, and dilution reaction chamber 140.

[0058] The analysis unit 700 calculates the toxicity index (TI) of the sample reaction vessel 130. s ) is required.

[0059] The toxicity index (TI) of such a sample reactor s ) is calculated by the following formula 1 to determine the correction coefficient of the standard reaction tank 120.

[0060]

number

[0061] [R0 = initial light intensity value after 1 minute of the mixed solution supplied from the bacterial dilution tank to the reference reaction tank, R1 = final light intensity value after 5 minutes of the mixed solution in the reference reaction tank after a fixed amount of dilution water has been added]

[0062] At this time, the analysis unit 700 determines that the calculated correction coefficient is valid if it is between 0.6 and 1.3, and if the correction coefficient is outside the range of 0.6 to 1.3, it conducts the toxicity evaluation test again with a new mixed liquid supplied again from the bacterial dilution tank 110 to each reaction tank.

[0063] Furthermore, the analysis unit 700 calculates the corrected light intensity value of the sample reaction vessel 130 to which a correction coefficient is applied using the following calculation formula 2. Here, the corrected light intensity value may be the light intensity value of the luminous bacteria to which the natural generation or decrease rate of the luminous bacteria is applied.

[0064]

number

[0065] [S0 = initial light intensity value after 1 minute of the mixture supplied from the bacterial dilution chamber to the sample reaction chamber, C0 = correction coefficient]

[0066] In addition, the analysis unit 700 calculates a toxicity index (TI) from the final light intensity value of the sample reaction chamber 130 measured after a predetermined time (5 minutes) has elapsed after a predetermined required sample is injected into the sample reaction chamber 130 from the sample supply unit 400 according to the following calculation formula 3: S ) is calculated.

[0067]

number

[0068] [S1 = corrected light intensity value after the first minute of the sample reaction chamber to which the correction coefficient has been applied, S2 = final light intensity value after a predetermined time has elapsed with a fixed amount of sample injected into the mixed solution in the sample reaction chamber]

[0069] If the toxicity index (TIs) of the sample reaction chamber 130 calculated by the analysis unit 700 is between 20 and 80%, the analysis unit 700 determines that a toxic substance is present in the sample. The analysis unit 700 calculates a correction coefficient based on the initial light intensity value of the reference reaction chamber 120 and the final light intensity value measured after a predetermined time (5 minutes) has elapsed, and quickly determines whether a toxic substance is present in the sample finally contained in the sample reaction chamber 130 based on the calculated correction coefficient. In this way, the ecotoxicity monitoring device 1000 according to the present invention can quickly determine whether a toxic substance is present in a sample more quickly than a conventional ecotoxicity monitoring device using luminescent bacteria.

[0070] Meanwhile, the analysis unit 700 calculates the toxicity index (TI) of the dilution reaction vessel 140 in the same manner as the sample reaction vessel 130. d ) is found.

[0071] The toxicity index (TI) of such a dilution reactor 140 d ) is obtained by using the correction coefficient of the reference reaction tank 120. Here, the correction coefficient of the reference reaction tank 120 has been described above, so a detailed description of it will be omitted.

[0072] Furthermore, the analysis unit 700 calculates the corrected light intensity value of the dilution reaction tank 140 to which a correction coefficient is applied using the following calculation formula 4. Here, the corrected light intensity value may be the light intensity value of the luminous bacteria to which the natural generation or decrease rate of the luminous bacteria is applied.

[0073]

number

[0074] [D0 = initial light intensity value after 1 minute of the mixed solution supplied from the bacterial dilution tank to the dilution reaction tank, C0 = correction coefficient]

[0075] In addition, the analysis unit 700 calculates the toxicity index (TI) from the final light intensity value of the dilution reaction tank 140 measured after the sample and dilution water are injected into the dilution reaction tank 140 at a required ratio from the sample supply unit 400 and dilution water supply unit 300 according to the following calculation formula 5, and after a predetermined time (5 minutes) has elapsed. d Here, the ratio of the sample to the dilution water injected into the dilution reaction tank 140 may be 1:1.

[0076]

number

[0077] [D1 = corrected light intensity value after the first minute of the dilution reaction vessel to which the correction coefficient has been applied, D2 = final light intensity value after a predetermined time has elapsed with the sample and dilution water injected into the mixture in the dilution reaction vessel]

[0078] The toxicity index (TI) of the dilution reaction tank 140 calculated through the analysis unit 700 is d ) is used to evaluate the toxicity risk of the sample. For example, the toxicity index (TI) of the sample reaction vessel 130 calculated by the analysis unit 700 is S ) at 50% of the toxicity index (TI d ) of the sample reaction vessel 130 calculated by the analysis unit 700 should be around 25%. S ) at 50% of the toxicity index (TI d ) is 40%, it means that a large number of luminous bacteria were killed even with a small amount of sample, and the sample can be judged to be highly toxic.

[0079] In this way, the analysis unit 700 calculates the toxicity index (TI S ) and the toxicity index (TI) of the dilution reactor 140 d ) and simultaneously evaluate the toxicity risk of the sample.

[0080] FIG. 5 is a flow chart showing a method for monitoring ecotoxicity using luminescent bacteria according to an embodiment of the present invention.

[0081] 5, a mixed solution of luminescent bacteria and dilution water is prepared in the bacteria dilution tank 110. The luminescent bacteria are supplied to the bacteria dilution tank 110 from the luminescent bacteria culturing unit 200, and dilution water is supplied from the dilution water supply unit 300 (S100).

[0082] Next, the mixture in the bacteria dilution tank 110 is supplied to the reference reaction tank 120, the sample reaction tank 130, and the dilution reaction tank 140, respectively (S200).

[0083] Next, after a predetermined time has elapsed, the optical measurement unit 500 measures the initial light intensity values ​​of the luminescent bacteria in the reference reaction chamber 120, the sample reaction chamber 130, and the dilution reaction chamber 140. Here, the predetermined time for the optical measurement unit 500 to measure the initial light intensity values ​​of the luminescent bacteria in the reference reaction chamber 120, the sample reaction chamber 130, and the dilution reaction chamber 140 may be one minute. The initial light intensity values ​​measured from each reaction chamber are provided to the analysis unit 700 (S300).

[0084] Next, after measuring the initial light intensity value, dilution water is supplied to the reference reaction chamber 120, a sample is supplied to the sample reaction chamber 130, and a predetermined amount of sample and dilution water is supplied to the dilution reaction chamber 140. The additional dilution water and sample supplied to the reference reaction chamber 120, sample reaction chamber 130, and dilution reaction chamber 140 may be supplied from the dilution water supply unit 300 and sample supply unit 400 (S400).

[0085] Next, after a predetermined time has elapsed, the optical measurement unit 500 measures the final light intensity values ​​of the luminescent bacteria in the reference reaction chamber 120, the sample reaction chamber 130, and the dilution reaction chamber 140. Here, the predetermined time may be 5 minutes. In this manner, the final light intensity values ​​measured from each reaction chamber are provided to the analysis unit 700 (S500).

[0086] Finally, the analysis unit 700 calculates a toxicity index using the initial and final light intensity values ​​of each reaction vessel measured by the optical measurement unit 500, and determines whether or not the sample is toxic based on the calculated toxicity index (S600).

[0087] Here, the analysis unit 700 calculates a correction coefficient based on the initial light intensity value and final light intensity value of the reference reaction chamber 120 measured through the optical measurement unit 500, calculates corrected light intensity values ​​of the sample reaction chamber 130 and the dilution reaction chamber 140 based on the calculated correction coefficient, and finally obtains the toxicity index of the sample reaction chamber 130 and the dilution reaction chamber 140.

[0088] However, this is merely one preferred embodiment of the present invention, and the scope of the present invention is not limited to the description of this embodiment.

[0089] The above description of the present invention is for illustrative purposes only, and those skilled in the art will understand that the present invention can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. Therefore, it should be understood that the above-described embodiments are illustrative in all respects and not limiting. For example, each component described as a single component may be implemented in a distributed form, and similarly, each component described as a distributed component may be implemented in a combined form.

[0090] The scope of the present invention is defined by the claims set forth below, and all modifications and variations that fall within the meaning and scope of the claims and their equivalent concepts should be construed as being included within the scope of the present invention. [Explanation of symbols]

[0091] 100 Reactor Modules 110 Bacteria dilution tank 120 Reference Reactor 130 Sample Reaction Tank 140 Dilution Reaction Tank 200 Luminous bacteria culture section 300 Dilution water supply section 400 Sample supply unit 500 Optical measurement section 510 Optical sensor unit 600 control section 700 Analysis Department 810 Casing 820 Monitor section 830 Waste Liquid Storage Department 1000 ecotoxicity monitoring device

Claims

1. a reaction chamber module including a reference reaction chamber, a sample reaction chamber, and a dilution reaction chamber, and a bacterial dilution chamber for supplying a mixture containing luminescent bacteria to the reference reaction chamber, the sample reaction chamber, and the dilution reaction chamber; a luminescent bacteria culturing unit for supplying luminescent bacteria to the bacteria dilution tank, the luminescent bacteria detecting the toxicity of the sample; a dilution water supply unit for supplying dilution water to the bacterial dilution tank, the reference reaction tank, and the dilution reaction tank; a sample supply unit that supplies a sample to the sample reaction vessel and the dilution reaction vessel; an optical measurement unit for measuring the amount of luminescence of luminescent bacteria in the toxicity evaluation substance provided from the reference reaction tank, the sample reaction tank to which the sample is supplied, and the dilution reaction tank; a control unit that controls the operation of the reaction vessel module, the luminescent bacteria culturing unit, the dilution water supply unit, the sample supply unit, and the optical measurement unit; an analysis unit that determines whether or not a toxic substance is present in a sample based on the light intensity value provided by the optical measurement unit; An ecotoxicity monitoring device using luminescent bacteria, comprising: the analysis unit calculates a correction coefficient based on the initial light intensity value of the reference reaction vessel measured by the optical measurement unit and the final light intensity value measured after a predetermined time has elapsed, and calculates corrected light intensity values ​​of the sample reaction vessel and the dilution reaction vessel based on the calculated correction coefficient; The analysis unit determines the toxicity index (TI s ) of the sample reaction vessel, The toxicity index (TI s ) of the sample reactor is: Calculating the correction coefficient of the reference reaction tank using the following calculation formula 1; calculating a corrected light intensity value of the sample reaction vessel to which the correction coefficient is applied by the following calculation formula 2; Calculating a toxicity index (TI s ) based on the final light intensity value of the sample reaction vessel measured after a predetermined amount of sample is injected into the sample reaction vessel and a predetermined time has elapsed, according to the following formula 3: characterized in that it is calculated by Biotoxicity monitoring device using luminescent bacteria: [Equation 1] R0 = initial light intensity value after 1 minute of the mixture delivered from the bacterial dilution chamber to the reference reaction chamber; R1 = final light intensity value after a predetermined time has elapsed with dilution water being injected into the mixture in the reference reaction tank; [Equation 2] S0 = initial light intensity value after 1 minute of the mixed solution supplied from the bacteria dilution chamber to the sample reaction chamber; C0 = correction factor; [Equation 3] S1 = corrected light intensity value after the initial 1 minute of the sample reaction vessel to which the correction coefficient was applied; S2 = Final light intensity value after a predetermined time has elapsed with a fixed amount of sample injected into the mixed solution in the sample reaction vessel.

2. The analysis unit calculates the toxicity index (TI) of the dilution reaction tank. ｄ ) but The toxicity index (TI ｄ )teeth, calculating a corrected light intensity value of the dilution reaction vessel to which the correction coefficient of the reference reaction vessel is applied by the following calculation formula 4; According to the following formula 5, the sample and dilution water are injected into the dilution reaction vessel at a required ratio from the sample supply unit and dilution water supply unit, and the toxicity index (TI) is calculated based on the final light intensity value of the dilution reaction vessel measured after a predetermined time has elapsed. ｄ ) and characterized in that it is calculated by The ecotoxicity monitoring device using the luminescent bacteria according to claim 1: [Equation 4] D0 = initial light intensity value after 1 minute of the mixed solution supplied from the bacterial dilution tank to the dilution reaction tank; C0 = correction factor; [Equation 5] D1 = corrected light intensity value after the first minute of the diluted reaction vessel to which the correction factor was applied; D2 = Final light intensity value after a predetermined time has elapsed with a fixed amount of sample and dilution water injected into the mixed solution in the dilution reaction tank.

3. The analysis unit If the calculated correction coefficient is between 0.6 and 1.3, it is determined to be valid. The calculated toxicity index (TI) of the sample reactor ｓ 3. The ecotoxicity monitoring device using luminescent bacteria according to claim 2, wherein it is determined that a toxic substance is present in the sample when the ratio of the luminescent bacteria to the total luminescent bacteria is 20 to 80%.

4. The analysis unit The toxicity index (TI ｓ ) and the toxicity index (TI ｄ 4. The ecotoxicity monitoring device using luminescent bacteria according to claim 3, wherein the toxicity risk of a sample is evaluated through a comparison of the luminescent bacteria.

5. 3. The ecotoxicity monitoring device using luminescent bacteria according to claim 2, wherein the predetermined time for measuring the final light intensity values ​​of the reference reaction chamber, the sample reaction chamber and the dilution reaction chamber is 5 minutes.

6. (A) preparing a mixture of luminescent bacteria and dilution water in a bacteria dilution tank; (B) supplying the mixture from the bacteria dilution tank to a reference reaction tank, a sample reaction tank, and a dilution reaction tank, respectively; (C) after a predetermined time has elapsed, measuring the initial light intensity values ​​of the luminescent bacteria in the reference reaction chamber, the sample reaction chamber, and the dilution reaction chamber through an optical measurement unit; (D) after measuring the initial light intensity value, supplying dilution water to the reference reaction chamber, supplying a sample to the sample reaction chamber, and supplying the sample and dilution water to the dilution reaction chamber in predetermined amounts; (E) after a predetermined time has elapsed, measuring the final light intensity values ​​of the luminescent bacteria in the reference reaction chamber, the sample reaction chamber, and the dilution reaction chamber through the optical measurement unit; (F) providing the initial light intensity value and the final light intensity value of each reaction vessel measured by the optical measurement unit to an analysis unit, which determines whether or not the sample is toxic; A method for monitoring ecotoxicity using luminescent bacteria, comprising: the analysis unit calculates a correction coefficient based on the initial light intensity value and the final light intensity value of the reference reaction vessel measured by the optical measurement unit, calculates corrected light intensity values ​​of the sample reaction vessel and the dilution reaction vessel based on the calculated correction coefficient, and obtains toxicity indices for the toxicity evaluation substance in the sample reaction vessel and the dilution reaction vessel; The analysis unit determines the toxicity index (TI s ) of the sample reaction vessel, The toxicity index (TI s ) of the sample reactor is: Calculating the correction coefficient of the reference reaction tank using the following calculation formula 1; calculating a corrected light intensity value of the sample reaction vessel to which the correction coefficient is applied by the following calculation formula 2; Calculating a toxicity index (TI s ) based on the final light intensity value of the sample reaction vessel measured after a predetermined amount of sample is injected into the sample reaction vessel and a predetermined time has elapsed, according to the following formula 3: characterized in that it is calculated by Ecotoxicity monitoring method using luminescent bacteria: [Equation 6] R0 = initial light intensity value after 1 minute of the mixture delivered from the bacterial dilution chamber to the reference reaction chamber; R1 = final light intensity value after a predetermined time has elapsed with dilution water being injected into the mixture in the reference reaction tank; [Equation 7] S0 = initial light intensity value after 1 minute of the mixed solution supplied from the bacteria dilution chamber to the sample reaction chamber; C0 = correction factor; [Equation 8] S1 = corrected light intensity value after the initial 1 minute of the sample reaction vessel to which the correction coefficient was applied; S2 = Final light intensity value after a predetermined time has elapsed with a fixed amount of sample injected into the mixed solution in the sample reaction vessel.

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