CN115046922A - Portable water quality comprehensive toxicity analyzer based on luminous bacteria and detection method - Google Patents
Portable water quality comprehensive toxicity analyzer based on luminous bacteria and detection method Download PDFInfo
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Abstract
The invention discloses a portable water quality comprehensive toxicity analyzer based on luminous bacteria and a detection method, wherein the portable water quality comprehensive toxicity analyzer comprises a detection bin unit, a photon counting data acquisition and photoelectric information conversion module, a power supply device, a light source and CCD array system, a software processing and system control module, an operation and display interface and a heat dissipation device; the detection bin unit is provided with four sample feeding channels; the photon counting data acquisition and photoelectric information conversion module acquires optical information and converts the optical information into electric information; the light source and the CCD array system quasi-convert the light information into image information. The portable water quality comprehensive toxicity analyzer realizes the conversion of the luminous bacteria detection method from laboratory operation to outdoor rapid detection; a CCD image acquisition sensor is additionally arranged, so that qualitative and quantitative analysis of organic pollutants in outdoor water is realized; by adopting a modern photoelectric detection means, the comprehensive toxicity and organic (micro) substance identification of the environmental water body can be effectively and rapidly monitored in the field.
Description
Technical Field
The invention relates to the technical field of water quality monitoring, in particular to a portable water quality comprehensive toxicity analyzer based on luminous bacteria.
Background
The luminous bacteria are organisms which can emit light in the normal metabolic process, most of the luminous bacteria live in the sea, only a few of the luminous bacteria live in the land or fresh water environment, all the luminous bacteria are gram-negative bacteria, most of the luminous bacteria are straight bacilli, and the few luminous bacteria are arc-shaped or club-shaped and have flagella. The luminous bacteria are tiny, the length is about 1.5-3.0 μm, the width is about 0.5- μm, and the wavelength range of the released blue-green fluorescence is 450-490 nm.
The luminescence mechanism of the luminous bacteria is the catalysis of luciferase of bacteria in the luminous bacteria, the luciferase is a generic name of enzymes catalyzing the oxidation luminescence of luciferin or fatty aldehyde in organisms, and molecular oxygen in the luminous bacteria is respectively oxidized into flavin nucleotide (FMN) and long-chain fatty acid by reduced flavin mononucleotide (FMH2) and long-chain fatty aldehyde (RCHO) under the catalysis of the bacterial luciferase, and simultaneously, the release of blue green light with the wavelength of 450-490 nm is accompanied. The light emitted by individual bacteria is extremely faint and barely visible to the naked eye, but can be detected by a corresponding luminescent bacteria detection device, such as a luminometer.
Foreign subjects inhibit bacterial luminescence primarily through the following two pathways: directly inhibiting the activity of enzymes participating in luminescence reaction; ② the metabolic process related to luminous reaction in cells is inhibited. Any toxic substance that interferes with or destroys the physiological processes of the luminescent bacteria such as respiration, growth, metabolism, etc. can be measured according to the change of the luminescence intensity. The detection mechanism of the luminescent bacteria is divided into specific detection and non-specific detection, the specific luminescent bacteria detection is that a special stress promoter is introduced into the upstream of a fluorescein gene according to the principle of a receptor-reporter gene (a 'lightson' mode), and when toxic substances exist in the growth environment of host cells, the toxic substances enter the host cells, induce a specific regulation and control element, further regulate and control the downstream gene expression and emit light sources. Generally, the monitoring of water quality belongs to non-specific detection, and the non-specific detection utilizes the principle of 'lightsoff' mode of luminescent bacteria, namely, extraneous toxic and harmful substances directly inhibit the activity of luciferase of the luminescent bacteria and inhibit physiological metabolism related to luminescent reaction in cells, so that a luminescent inhibition effect is formed on the luminescent bacteria. In the "lightsoff" assay, sample toxicity is estimated by the degree of inhibition of normal cellular activity, which can occur at any stage of the reaction or at any point affecting cell growth and development. Bioluminescence is widely dependent on cell metabolism and needs high-energy cofactors, so when the bioluminescence is contacted with external toxic and harmful substances, the activity of cells is reduced, the ATP content level is reduced, and the luminous intensity of luminous bacteria is reduced.
A water toxicity analyzer in the market generally comprises a photoelectric detection device, a data processing part and a data display part, and mainly comprises an indoor test type and a portable type. The two types of water toxicity analyzers have different testing characteristics, and an indoor testing type is large in size but high in measuring precision; the portable body size is small but the measurement precision is low, and the indoor test type is still required for high-precision data measurement.
Disclosure of Invention
In order to solve the technical problems, the invention provides a portable water quality comprehensive toxicity analyzer based on luminous bacteria and a detection method.
The technical scheme adopted by the invention for solving the technical problems is as follows: portable water quality comprehensive toxicity analyzer based on luminous bacteria comprises:
a detection bin unit: for storing the test sample;
photon counting data acquisition and photoelectric information conversion module: the device is used for measuring the luminous intensity of a water sample to be measured;
light source and CCD array system: the qualitative and quantitative data are used for measuring the organic pollutants in the water sample to be measured;
a system control module: for processing analytical measurement data.
Preferably, the detection bin unit comprises a first detection cell, a second detection cell, a third detection cell and a fourth detection cell.
Preferably, the photon counting data acquisition and photoelectric information conversion module comprises a photon counting data acquisition unit, a photoelectric information processor and a photomultiplier tube.
Preferably, the light source and CCD array system includes a two-pin LED lamp, a flat-field concave holographic grating, and a CCD image sensor.
Preferably, the system control module comprises a raspberry type microcomputer.
The detection method of the portable water quality comprehensive toxicity analyzer based on the luminous bacteria is characterized by comprising the following steps: the detection process of the water sample to be detected comprises the following steps:
taking a bottle of the photobacterium brightens dry powder, standing for 10min at room temperature, adding 500 microliters of resuscitation diluent, standing for 15min, adding 100 microliters of osmotic pressure regulating fluid into a cuvette, then adding 900 microliters of water sample to be detected, adding resuscitation bacteria solution, timing for 5min, 10min and 15min, and then carrying out photon counting detection.
Preferably, during the water quality analysis process,
the optical information of the luminous bacteria of the mobile phone is processed into the luminous inhibition rate and the corresponding comprehensive toxicity level of the water quality through a photon technology data collector;
and acquiring the light information of the water sample through the CCD image sensor, and processing the light information into corresponding image information.
Preferably, in the water quality detection process, after a water sample to be detected is placed in the detection bin, the photon technology data collector starts to time; when the timing reaches t, detecting the luminous intensity of the sample, and calculating the relative luminous inhibition rate after the reaction time t, wherein the calculation formula is as follows: h is 100 × (1-Ist/Ict); and judging the comprehensive toxicity of the water sample to be detected according to the luminescence inhibition rate.
Preferably, starting a photon counting data acquisition negative detection process, replacing a water sample to be detected with sterile water, repeatedly executing the water quality detection process, and checking the relative luminescence inhibition rate H; if the relative luminescence inhibition rate H is within +/-10%, judging that the luminescent bacteria in the bacterial liquid are normal and the parallelism of the analyzer meets the requirement; otherwise, replacing the bacterial liquid or detecting whether the analyzer has a fault.
Preferably, a positive detection process is started, the water sample to be detected is replaced by the positive quality control liquid, the water quality detection process is repeatedly executed, and the relative luminescence inhibition rate H is checked; if the relative luminescence inhibition rate H is within 50% +/-10%, judging that the luminescent bacteria in the bacterial liquid are normal and the parallelism of the analyzer meets the requirement; otherwise, replacing the bacterial liquid or detecting whether the analyzer has a fault.
The invention has the advantages that: the analyzer realizes a high-precision portable luminous bacteria detection method, improves the portability of the analyzer by adopting a photoelectric detection means and optimizing internal components of the existing water quality comprehensive toxicity analyzer and the additionally arranged CCD image acquisition and sensing function, simplifies the operation, realizes high-precision monitoring of comprehensive toxicity of heavy metal, organic pollutants, pesticide residues, antibiotics and the like in environmental water, realizes qualitative and quantitative analysis of organic pollutants in field water, is particularly suitable for being applied to a field water source place and a water plant entrance and exit, and is also suitable for laboratory detection.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments will be briefly introduced below, it is obvious that the drawings in the following description are only two of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without creative efforts.
FIG. 1 is a block diagram of an overall architecture of an embodiment of the present invention;
FIG. 2 is a left side view of FIG. 1 of an embodiment of the present invention;
the system comprises a detection bin unit 100, a photon counting data acquisition and photoelectric information conversion module 200, a power supply device 300, a light source and CCD array system 400, a heat dissipation device 500, a software processing and system control module 600, an operation and display interface 700, a first detection cell 101, a second detection cell 102, a third detection cell 103, a fourth detection cell 104, a photoelectric information processing unit 201, a photomultiplier 202, a rechargeable mobile cell 301, a transformer 302, a light source 401, a flat field concave holographic grating 402 and a CCD image sensor 403.
Detailed Description
For the purpose of enhancing understanding of the present invention, the present invention will be described in further detail with reference to the accompanying drawings and examples, which are provided for illustration only and are not intended to limit the scope of the present invention.
Examples
As shown in figure 1, the scheme optimizes core components by using a luminous bacteria detection method and existing market devices, converts a large-scale detection analyzer into a portable type, adds a CCD array system 400, realizes qualitative and quantitative analysis of field rapid detection of organic pollutants in water, and can rapidly and effectively monitor comprehensive toxicity of environmental water in the field.
A portable water quality comprehensive toxicity analyzer based on luminous bacteria comprises a detection bin unit 100, a photon counting data acquisition and photoelectric information conversion module 200, a power supply device 300, a light source 401, a CCD array system 400, a software processing and system control module 600, an operation and display interface 700 and a heat dissipation device 500.
The detection bin unit 100 is used for storage and detection of a detection water sample, a negative water sample and a positive water sample, and consists of a first detection pool, a second detection pool, a third detection pool and a fourth detection pool. The detection bin unit 100 can slide through a pull rod, a first channel is a negative control, a second channel is a positive control, a third channel and a fourth channel are detection water samples, and every ten detection samples need to be provided with the positive control and the negative control;
the photon counting data acquisition and photoelectric information conversion module 200 comprises a photon counting data acquisition unit, a photoelectric information processor and a photomultiplier 202202, wherein the photon counting data acquisition unit is used for collecting optical information of luminous bacteria, the photoelectric information processor is used for converting the optical information into electric information and amplifying the electric information, the photomultiplier 202202 is used for detecting the luminous intensity of a water sample to be detected, and the photoelectric information processing unit 201 is used for processing and converting the optical information and the electric information;
the system comprises a light source 401 and a CCD array system 400, wherein the light source 401 provides strong light for a CCD image sensor 403, and the CCD image sensor 403 detects substances absorbing light in a sample water sample under the strong light and converts the substances into electric signals; the power supply apparatus 300 includes a 12V rechargeable battery and a transformer 302, the 12V rechargeable battery provides power to the whole apparatus, and the transformer 302 is used for changing the battery voltage to provide power of corresponding voltage for each primary component. The device comprises two needle type LED lamps, a flat field concave holographic grating 402 and a CCD image sensor 403, wherein the two needle type LED lamps are used for providing strong light, the flat field concave holographic grating 402 is used for controlling the wavelength of incident light to be stably input between 200 nm and 800nm, and the CCD image sensor 403 is used for detecting light absorption information of light absorption substances in a water sample to be detected.
The software processing and system control module 600 comprises a raspberry microcomputer, wherein the raspberry microcomputer stores the set photon counting data processing software, and can process the collected photoelectric information of the luminous bacteria into the luminous inhibition rate and the corresponding water quality biological comprehensive toxicity level; the set CCD image processing software can process and convert the collected electric signals of the CCD image sensor 403 into corresponding image information;
an operation and display interface 700 for touch screen operation using instrument functions; the heat dissipation device 500 includes a heat dissipation fan for dissipating heat of the whole machine, so as to ensure a proper temperature during the use of the whole machine.
The portable analyzer also comprises photon counting data processing software designed in a software processing and system control module 600, the software is mainly divided into two plates, the first plate is a detection unit, the second plate is an analysis processing unit, the detection unit controls the whole process of detecting the luminous intensity of luminous bacteria in a water sample to be detected by the whole analyzer, and the analysis processing unit is used for analyzing and calculating the relative inhibition luminous rate and the corresponding toxicity level; the analyzer further includes CCD image processing software designed in the software processing and system control module 600, which is used to control the overall process of the CCD image sensor 403 for detecting the water sample to be tested.
The detection process of the water sample to be detected comprises the following steps:
taking a bottle of bright photobacterium dry powder, standing at room temperature for 10min, adding 500 microliters of recovery diluent, standing for 15min, adding 100 microliters of osmotic pressure regulating solution into a cuvette, then adding 900 microliters of water sample to be detected, adding recovery bacterial solution, timing for 5, 10 and 15min, and then carrying out photon counting detection.
In the water quality analysis process, the software processing and control module executes the following water quality detection process;
controlling the photon counting data collector to collect light information of the luminous bacteria, transmitting the light information to the photoelectric information processor, converting the light information into electric information and amplifying the electric information;
controlling the light intensity of the light source 401, collecting light information of a water sample by the CCD image sensor 403, converting the light information into an electric signal, and converting the electric signal into image information in the software processing module;
controlling the photon counting data processing software to process the collected photoelectric information of the luminous bacteria into a luminous inhibition rate and a corresponding comprehensive toxicity level of water quality;
controlling the CCD image processing software to process and convert the collected electric signals of the CCD image sensor 403 into corresponding image information;
in the water quality detection process, after a water sample to be detected is detected by the detection bin unit 100, the control module selects to start the photon counting data collector and starts timing, the photon counting data collector collected by the signal collection module detects the luminous intensity of the detected sample when the timing reaches t, photon counting data processing software arranged in the software processing module calculates the relative luminous inhibition rate after the reaction time t by using a formula H (%) -100 x (1-Ist/Ict), and the photon counting data processing software judges the comprehensive toxicity of the water sample to be detected by using a table 1 according to the relative luminous inhibition rate; wherein Ist is the luminous intensity of the water sample to be detected in the sample detection bin; ict is the luminous intensity of the negative water sample in the reference detection bin.
Relative luminescence inhibition percent toxicity rating
In the water quality detection process, the control module selectively starts the light source 401 and the CCD array system 400 after the water sample to be detected is detected by the detection chamber unit 100, the light with different wavelengths passes through the water sample to be detected, the CCD image sensor 403 collects the light information passing through the water sample to be detected and converts the light information into an electric signal, and the CCD image processing software processes and converts the collected electric signal of the CCD image sensor 403 into corresponding image information.
The control module selectively initiates a photon counting data acquisition negative detection process:
starting a negative detection process, replacing the water sample to be detected with sterile water, repeatedly executing the water quality detection process, and checking the relative luminescence inhibition rate H (%); if the relative luminescence inhibition rate H (%) is within +/-10%, judging that the luminescent bacteria in the bacterial liquid are normal and the parallelism of an analyzer meets the requirement; otherwise, replacing the bacterial liquid or detecting whether the analyzer has a fault;
the control module selectively initiates a photon counting data acquisition positive detection process:
starting a positive detection process, replacing the water sample to be detected with a positive quality control solution, repeatedly executing the water quality detection process, and checking the relative luminescence inhibition rate H (%); if the relative luminescence inhibition rate H (%) is within 50% +/-10%, judging that the luminescent bacteria in the bacterial liquid are normal and the parallelism of an analyzer meets the requirement; otherwise, replacing the bacteria liquid or detecting whether the analyzer has a fault.
Further comprising: the heat dissipation device 500 comprises a heat dissipation fan, and the fan dissipates heat and cools the software processing and system control module 600 in the operation process of the portable water quality comprehensive toxicity analyzer, so that a card machine is prevented from being blocked due to high temperature; the power supply equipment 300 comprises a battery and a transformer 302, wherein the battery provides operation energy for the portable water quality comprehensive toxicity analyzer, and the transformer 302 regulates the voltage of the battery and provides different voltage power supplies for each element in the analyzer; a charging hole for providing an external power supply to the battery; the pull rod does the detection bin provides a sliding function to detect water samples to be detected in each detection pool in the detection bin.
The sample detection pool and the reference detection pool in the detection bin are both hollow vertical rectangles, a light source 401 is arranged beside one vertical surface of the vertical rectangle detection pool, the other side of the vertical surface provided with the light source 401 is a photon counting data collector and a CCD image sensor 403, a sliding cover is arranged on the top surface of the vertical rectangle detection pool, and an outlet for placing and taking out a water sample to be detected is formed.
In some embodiments of the present application, the control module preferably performs the following water quality detection process in the water quality integrated toxicity analysis process:
taking a bottle of bright photobacterium dry powder, standing for 10min at room temperature, adding 500 microliters of resuscitation diluent, uniformly mixing, standing for 15min, adding 100 microliters of osmotic pressure regulating fluid into the cuvette, sequentially adding 900 microliters of water samples to be detected into the cuvette, finally adding 50 microliters of bacteria solution after resuscitation into each cuvette, starting timing from adding the bacteria solution into the first cuvette, and after all the bacteria solution is added, rapidly placing the cuvettes into four detection tanks of a detection bin of the portable water quality comprehensive toxicity analyzer in sequence.
Controlling the operation and display interface 700 to provide instructions for the software processing and control module, sending measurement instructions to the photon counting data collector by the software processing and control module, measuring the luminous intensity of the water sample to be detected in each detection pool in the detection bin, and automatically transmitting the obtained data to the software processing and control module;
the software processing and control module is controlled to carry out data analysis by built-in processing software, the analysis structure comprises the relative light-emitting inhibition rate and the comprehensive toxicity level, and the data is displayed and output on the operation and display interface 700.
In some embodiments of the present application, the control module preferably performs the following water quality detection process in the water quality CCD image analysis process:
the operation and display interface 700 is controlled to provide instructions for the software processing and control module, the software processing and control module sends a starting instruction to the light source 401, the flat-field concave holographic grating 402 is controlled to regulate and control the incident light to be a set wavelength, the CCD image acquisition sensor measures organic pollutants of water samples to be detected in each detection pool in the detection bin, mainly spectral characteristics of the organic pollutants under different wavelengths, and the obtained data are automatically transmitted to the software processing and control module;
and controlling processing software built in the processing and control module to perform data analysis, wherein the analysis result comprises the image resolution of the organic pollutants of the water sample to be detected under the condition of receiving optical signals with different wavelengths, and displaying and outputting the image resolution on the operation and display interface 700.
Compared with the prior art, the invention has the advantages and positive effects that: the analyzer realizes a high-precision portable luminous bacteria detection method, adopts a photoelectric detection means and designs automatic data processing analysis software, thereby realizing portable, automatic and high-efficiency monitoring of water quality comprehensive toxicity. Through optimizing the internal fittings of the existing water quality comprehensive toxicity analyzer, designing data collection and processing software and additionally arranging a CCD image acquisition and sensing function, the portability and the automation degree of the analyzer are improved, the operation is simplified, the high-precision monitoring on the comprehensive toxicity of heavy metals, organic pollutants, pesticide residues, antibiotics and the like in an environmental water body is realized, the qualitative and quantitative analysis on the organic pollutants in a field water body is realized, and the method is particularly suitable for being applied to a field water source area and a water plant entrance and exit and is also suitable for laboratory detection.
The above embodiments should not limit the present invention in any way, and all technical solutions obtained by using equivalent alternatives or equivalent transformations fall within the protection scope of the present invention.
Claims (10)
1. Portable water quality comprehensive toxicity analyzer based on luminous bacteria is characterized by comprising:
a detection bin unit: for storing the test sample;
photon counting data acquisition and photoelectric information conversion module: the device is used for measuring the luminous intensity of a water sample to be measured;
light source and CCD array system: the qualitative and quantitative data are used for measuring the organic pollutants in the water sample to be measured;
a system control module: for processing analytical measurement data.
2. The portable water quality comprehensive toxicity analyzer based on luminous bacteria of claim 1, characterized in that: the detection bin unit comprises a first detection pool, a second detection pool, a third detection pool and a fourth detection pool.
3. The portable water quality comprehensive toxicity analyzer based on luminous bacteria of claim 1, characterized in that: the photon counting data acquisition and photoelectric information conversion module comprises a photon counting data acquisition unit, a photoelectric information processor and a photomultiplier.
4. The portable water quality comprehensive toxicity analyzer based on luminous bacteria of claim 1, characterized in that: the light source and CCD array system comprises two needle type LED lamps, a flat field concave holographic grating and a CCD image sensor.
5. The portable water quality comprehensive toxicity analyzer based on luminous bacteria of claim 1, characterized in that: the system control module comprises a raspberry type microcomputer.
6. The method for detecting a portable water quality comprehensive toxicity analyzer based on luminous bacteria of any one of claims 1 to 5, wherein: the detection process of the water sample to be detected comprises the following steps:
taking a bottle of the photobacterium brightens dry powder, standing for 10min at room temperature, adding 500 microliters of resuscitation diluent, standing for 15min, adding 100 microliters of osmotic pressure regulating fluid into a cuvette, then adding 900 microliters of water sample to be detected, adding resuscitation bacteria solution, timing for 5min, 10min and 15min, and then carrying out photon counting detection.
7. The method for detecting the portable water quality comprehensive toxicity analyzer based on the luminous bacteria as claimed in claim 6, wherein the method comprises the following steps: in the process of analyzing the water quality,
the optical information of the luminous bacteria of the mobile phone is processed into the luminous inhibition rate and the corresponding comprehensive toxicity level of the water quality through a photon technology data collector;
and acquiring the light information of the water sample through the CCD image sensor, and processing the light information into corresponding image information.
8. The method for detecting the portable water quality comprehensive toxicity analyzer based on the luminous bacteria as claimed in claim 6, wherein the method comprises the following steps: in the water quality detection process, after a water sample to be detected is placed in a detection bin, a photon technology data collector starts to time; when the timing reaches t, detecting the luminous intensity of the sample, and calculating the relative luminous inhibition rate after the reaction time t, wherein the calculation formula is as follows: h100 × (1-Ist/Ict); and judging the comprehensive toxicity of the water sample to be detected according to the luminescence inhibition rate.
9. The method for detecting the portable water quality comprehensive toxicity analyzer based on the luminous bacteria as claimed in claim 6, wherein the method comprises the following steps: starting a photon counting data acquisition negative detection process, replacing a water sample to be detected with sterile water, repeatedly executing the water quality detection process, and checking the relative light emission inhibition rate H; if the relative luminescence inhibition rate H is within +/-10%, judging that the luminescent bacteria in the bacterial liquid are normal and the parallelism of the analyzer meets the requirement; otherwise, replacing the bacteria liquid or detecting whether the analyzer has a fault.
10. The method for detecting the portable water quality comprehensive toxicity analyzer based on the luminous bacteria as claimed in claim 6, wherein the method comprises the following steps: starting a positive detection process, replacing the water sample to be detected with a positive quality control solution, repeatedly executing the water quality detection process, and checking the relative light emission inhibition rate H; if the relative luminescence inhibition rate H is within 50% +/-10%, judging that the luminescent bacteria in the bacterial liquid are normal and the parallelism of the analyzer meets the requirement; otherwise, replacing the bacteria liquid or detecting whether the analyzer has a fault.
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