CN108362532A - A kind of method that osmotic pressure adjusts liquid and carries out poisonous substance detection using it - Google Patents
A kind of method that osmotic pressure adjusts liquid and carries out poisonous substance detection using it Download PDFInfo
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- CN108362532A CN108362532A CN201711446889.6A CN201711446889A CN108362532A CN 108362532 A CN108362532 A CN 108362532A CN 201711446889 A CN201711446889 A CN 201711446889A CN 108362532 A CN108362532 A CN 108362532A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
- G01N21/763—Bioluminescence
Abstract
The present invention relates to the water quality monitoring fields in environmental protection industry (epi), the present invention provides a kind of osmotic pressure to adjust liquid, it includes NaCl, Hepes, magnesium compound, DMSO and suitable deionized water, the NaCl contents are 20wt% 30wt%, the Hepes contents are 10wt% 20wt%, the magnesium compound content is 2.0wt% 5.0wt%, and the DMSO contents are 5% 20% (V/V)；The method that liquid carries out poisonous substance detection is adjusted using osmotic pressure the present invention also provides a kind of, liquid is adjusted by the way that a certain proportion of osmotic pressure is added into sample solution, improve the luminous intensity of photobacteria, and detection sensitivity of the photobacteria to the poisonous substances such as insoluble organic matter, low-concentration organic, low solubility organic matter and other heavy metals in water is improved, and then promote Luminous bacteria in the application range of water quality comprehensive toxicity.
Description
【Technical field】
The present invention relates to the water quality monitoring field in environmental protection industry (epi), more particularly to a kind of osmotic pressure adjust liquid and using its into
The method of row poisonous substance detection.
【Background technology】
With the high speed development of economic society, China's shortage of water resources problem getting worse, however the acute dirt of China's water body
Dye event frequently occurs, and how quickly to detect water pollution situation becomes the significant challenge of field of water quality detection.And photobacteria
Method is suitable for industrial wastewater, the acute toxicity of water quality monitoring of soluble chemical substance under sewage body and laboratory condition received.
In actual application, different for the reaction susceptibility of different toxicants due to photobacteria, and work as
When the concentration of test substance is too low, due to the iris action of cell membrane, it is long that toxicant enters action time in bacterial body,
Inhibition can not be showed in short time.
Thus, it is badly in need of a kind of method improving photobacteria susceptibility, expands the scope of application of photobacteria.
【Invention content】
To overcome the problems, such as that existing Luminous bacteria poisonous substance detection sensitivity is low.The present invention provides a kind of adjustings of osmotic pressure
Liquid and a kind of method carrying out poisonous substance detection using it.
The present invention solves above-mentioned technical problem, and the technical solution provided is as follows：A kind of osmotic pressure adjusting liquid comprising
NaCl, Hepes, magnesium compound, DMSO and suitable deionized water, the NaCl contents are 20wt%-30wt%, described
Hepes contents are 10wt%-20wt%, the magnesium compound content is 2.0wt%-5.0wt%, and the DMSO contents are 5%-
20% (V/V).
Preferably, the NaCl contents be 20wt%, Hepes content be 15wt%, magnesium compound content be 4.0wt% and
DMSO contents are 10% (V/V).
Preferably, it further includes PH conditioning agents that the osmotic pressure, which adjusts liquid,.
Preferably, the PH conditioning agents are the NaOH solution of 1mol/L.
Preferably, the osmotic pressure adjusts the PH ranging from 7.4-8.0 of liquid.
Preferably, the magnesium compound includes MgCl2、MgSO4、Mg(NO3)2Or Mg (OH)2One or more of.
The present invention solves above-mentioned technical problem, and the technical solution provided is as follows：It is a kind of to adjust liquid progress using osmotic pressure
The method of poisonous substance detection comprising：
Recovery photobacteria, to obtain recovery photobacteria bacterium solution；
Osmotic pressure as described above is provided and adjusts liquid, and osmotic pressure is adjusted into liquid and is added in sample solution and contrast solution, institute
It is (1.5-0.5) to state osmotic pressure and adjust liquid and the sample solution mixed volume ratio：(8.5-9.5), the osmotic pressure are adjusted
Liquid and the contrast solution mixed proportion and the osmotic pressure adjust liquid and the sample solution mixed volume ratio is identical, to obtain
Obtain pretreatment sample solution and pretreatment control solution；
Recovery photobacteria solution is added in pretreatment sample solution and pretreatment control solution, and every milliliter of sample is molten
The recovery photobacteria bacterium solution of 8-15 μ L, and isothermal holding certain time at a certain temperature are separately added into liquid or contrast solution
Sample to be tested and control to be checked are obtained afterwards；
The sample to be tested and control to be checked are detected, to obtain corresponding detection data.
Preferably, it is 1 that the osmotic pressure, which adjusts liquid and the volume ratio in the sample solution or contrast solution is added,：9.
Preferably, the recovery photobacteria bacterium solution of 10 μ L is separately added into every milliliter of sample solution or contrast solution.
Preferably, the temperature is 20 DEG C, and the isothermal holding time is 15min or 30min.
Compared with prior art, osmotic pressure provided by the present invention adjusts liquid, by adding magnesium compound, makes osmotic pressure tune
Mg is contained in section liquid2+, and by controlling Mg2+Its content effectively enhances the fluorescein for participating in photobacteria luminescence-producing reaction
The activity of enzyme, and then enhance the luminous intensity of photobacteria；Meanwhile the osmotic pressure adjusts liquid by adding Hepes and fitting
The NaOH solution of the 1mol/L of amount has adjusted the osmotic pressure and adjusts the pH value of liquid, and then the osmotic pressure is avoided to adjust liquid
PH value impacts sample P H values；Further, the osmotic pressure adjusts liquid, by being added to photobacteria hypotoxicity
DMSO solution can effectively increase the solubility of organic matter, it is particularly possible to improve insoluble in water, low solubility organic matter
And the solubility of low content organic matter, and DMSO solution can enhance the permeability of light-emitting bacterial cells film, make poisonous substance more
It is easily accessible inside photobacteria, and then improves photobacteria to insoluble organic matter, low-concentration organic, low dissolving in water
The detection sensitivity for spending the poisonous substances such as organic matter and other heavy metals makes osmotic pressure adjust liquid and promotes Luminous bacteria comprehensive in water quality
Close the application range of toxicity.
It is provided by the present invention that the method that liquid carries out poisonous substance detection is adjusted using osmotic pressure, by sample solution and control
A certain proportion of osmotic pressure is sequentially added in solution and adjusts liquid, recovery photobacteria solution, makes photobacteria when detecting sample,
With better luminous intensity and cell permeability, poisonous substance is set to be more easy to enter in photobacteria body faster, final photobacteria
Stronger luminous inhibiting rate is shown under identical concentration of poison, i.e. photobacteria only needs smaller concentration of poison, so that it may
To obtain identical luminous inhibiting rate, sensitivity of the photobacteria to poisonous substance is improved；Meanwhile by sample with compare in plus
Enter osmotic pressure and adjust liquid, makes poisonous substance, the low solubility organic matter of low concentration, especially improve the low solubilities organic matter such as pesticide
Solubility, further expand the detection sensitivity and application range of photobacteria poisonous substance detection method.
【Description of the drawings】
Fig. 1 is a kind of method flow signal adjusting liquid progress poisonous substance detection using osmotic pressure in second embodiment of the invention
Figure.
Fig. 2 is shone carefully in a kind of method using osmotic pressure adjusting liquid progress poisonous substance detection in second embodiment of the invention
Bacterium recovery flow diagram.
Fig. 3 is a kind of method flow signal adjusting liquid progress poisonous substance detection using osmotic pressure in third embodiment of the invention
Figure.
Fig. 4 is Mg in the first specific implementation mode of the invention2+Content is to photobacteria luminous intensity testing result schematic diagram.
Fig. 5 is that DMSO contents shine the signal of inhibiting rate testing result to photobacteria in second specific implementation mode of the invention
Figure.
Fig. 6 is that the osmotic pressure added in the present invention in 30%NaCl solution and the addition present invention adjusts liquid, to different content
Hg2+Poisonous substance sample is detected, the luminous inhibiting rate testing result contrast schematic diagram of corresponding photobacteria.
Fig. 7 is that the osmotic pressure added in the present invention in 30%NaCl solution and the addition present invention adjusts liquid, to different content
3,5- Dichlorophenols poisonous substance sample is detected, the luminous inhibiting rate testing result contrast schematic diagram of corresponding photobacteria.
Fig. 8 is that the osmotic pressure added in the present invention in 30%NaCl solution and the addition present invention adjusts liquid, to different content
Decis poisonous substance sample is detected, the luminous inhibiting rate testing result contrast schematic diagram of corresponding photobacteria.
Fig. 9 is that the osmotic pressure added in the present invention in 30%NaCl solution and the addition present invention adjusts liquid, to different content
Nitrofen poisonous substance sample is detected, the luminous inhibiting rate testing result contrast schematic diagram of corresponding photobacteria.
【Specific implementation mode】
In order to make the purpose of the present invention, technical solution and advantage be more clearly understood, below in conjunction with attached drawing and embodiment,
The present invention will be described in further detail.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention,
It is not intended to limit the present invention.
In the first embodiment of the present invention, provide a kind of osmotic pressure and adjust liquid, the osmotic pressure adjust liquid include NaCl,
Hepes (4- (2-Hydroxyethyl) piperazine-1-ethanesulfonic acid, 4- hydroxyethyl piperazineethanesulfonic acid),
Magnesium compound, DMSO (Dimethyl sulfoxide, dimethyl sulfoxide) and suitable deionized water, the Hepes contents are
10wt%-20wt%, magnesium compound content are 1.0wt%-5.0wt%, and NaCl contents are 20wt%-30wt%, the DMSO
Content is 5%-20% (V/V)；
Further, the Hepes contents can also be 12wt%-18wt%；Specifically, the Hepes contents can also be
12wt%, 14wt%, 16wt% or 18wt%.The magnesium compound content is 1.5wt%-4.5wt%；Specifically, the magnesium
Compounds content be 1.0wt%, 1.5wt%, 2.0wt%, 2.5wt%, 3.0wt%, 3.5wt%, 4.0wt%, 4.5wt% or
5.0wt%.The NaCl contents are 22wt%-28wt%；Specifically, the NaCl contents be 20wt%, 22wt%,
25wt%, 28wt% or 30wt%.Further, the DMSO contents are 8%-18% (V/V)；Specifically, the DMSO contains
Amount is 5%, 8%, 10%, 15%, 18% or 20%.
Further, NaCl contents described in the present embodiment is preferably that 20wt%, the Hepes contents are preferably
15wt%, the magnesium compound content are preferably 4.0wt% and the DMSO contents are preferably 10% (V/V).
Further, it further includes PH conditioning agents that the osmotic pressure, which adjusts liquid, and the osmotic pressure regulator is 1mol/L's
NaOH solution, the osmotic pressure adjust the pH range of liquid in 7.4-8.0；Specifically the pH value of the osmotic pressure adjusting liquid is
7.4,7.6,7.8 or 8.0；The pH value that osmotic pressure described in the present embodiment adjusts liquid is preferably 7.8.
Wherein, the magnesium compound is mainly that the osmotic pressure adjusts liquid offer Mg2+, the magnesium compound includes MgCl2、
MgSO4、Mg(NO3)2Or Mg (OH)2One or more of；Further, magnesium compound described in the present embodiment is preferably
MgCl2And/or MgSO4。
Further, in the present invention, the osmotic pressure adjusts liquid and can be prepared as follows acquisition：
It is first that the magnesium compound of NaCl, 2.0wt%-5.0wt% of 20wt%-30wt% and appropriate amount of deionized water is advanced
After row mixing, the DMSO of 5%-20% (V/V) is added, the Hepes and suitable 1mol/L of 10wt%-20wt% are eventually adding
NaOH solution, after the PH of the mixed solution is adjusted to 7.4-8.0, obtains the osmotic pressure and adjust liquid.
Referring to Fig. 1, in the second embodiment of the present invention, provides and a kind of utilize osmotic pressure to adjust liquid to carry out poisonous substance detection
Method S10, include mainly：
Step S11, recovery photobacteria, to obtain recovery photobacteria bacterium solution；
Step S12 provides the osmotic pressure described in above-mentioned first embodiment and adjusts liquid, and osmotic pressure adjusted liquid, sample is added
In product solution and contrast solution, the osmotic pressure adjusts liquid and the sample solution mixed volume ratio is (1.5-0.5)：
(8.5-9.5), the osmotic pressure adjust liquid and adjust liquid and the sample with the contrast solution mixed proportion and the osmotic pressure
Solution mixed volume ratio is identical, to obtain pretreatment sample solution and pretreatment control solution.
Recovery photobacteria solution is added in pretreatment sample solution and pretreatment control solution step S13, and per milli
Rise the recovery photobacteria bacterium solution that 8-15 μ L are separately added into sample solution or contrast solution, and isothermal holding at a certain temperature
Sample to be tested and control to be checked are obtained after a certain period of time；
Step S14 is detected the sample to be tested and control to be checked, to obtain corresponding detection data.
It is appreciated that the Hepes contents for including in the pretreatment sample solution and pretreatment control solution are 1.0wt%-
2.0wt%, Mg2+Content is 0.2wt%-0.6wt%, and NaCl contents are 2.0wt%-3.0wt%, the DMSO and osmotic pressure tune
The volume ratio for saving liquid is 0.5%-2.0% (V/V).
Referring to Fig. 2, further, in the step S11, the photobacteria is photobacterium phosphoreum, is recovered
Photobacteria solution is specifically further comprising the steps of：
S111 provides photobacterium phosphoreum freeze-dried powder；
S112 provides 3%NaCl resuscitation fluids；
S113 moves into resuscitation fluid in photobacterium phosphoreum freeze-dried powder, and adds in every gram of photobacterium phosphoreum freeze-dried powder
Enter 1mL3%NaCl resuscitation fluids, resuscitation fluid is waited for obtain photobacterium phosphoreum；
Photobacterium phosphoreum is waited for resuscitation fluid at 15-25 DEG C by S114, isothermal holding 15min, to obtain bright burn bar
Bacterium resuscitation fluid.
Further, in the step S13, the osmotic pressure adjusts liquid and the sample solution mixed volume ratio is specific
Can be 1.5:8.5、1.5:9.0、1.5:9.5、1.0:8.5、1.0:9.0、1.0:9.5、0.5:8.5、0.5:9.0 or 0.5:
9.5；It is preferably 1.0 that osmotic pressure described in the present embodiment, which adjusts liquid and the sample solution mixed volume ratio,:9.0.
In the step S13, in every milliliter of pretreatment sample solution and pretreatment control solution, recovery photobacteria is added
The volume of solution is specifically as follows 8 μ L, 9 μ L, 10 μ L, 12 μ L or 15 μ L, in the present embodiment every milliliter of pretreatment sample solution and
In pretreatment control solution, the volume that recovery photobacteria solution is added is preferably 10 μ L.And the temperature in the S13 is 20
DEG C, soaking time is 15min or 30min.
Referring to Fig. 3, in third embodiment of the invention, provides and a kind of adjusting liquid using osmotic pressure and carry out poisonous substance detection
Method S20, specifically includes：
S21, provides 0.5g photobacterium phosphoreums freeze-dried powder and 3%NaCl resuscitation fluids, and to photobacterium phosphoreum freeze-dried powder
Middle addition 5mL3%NaCl resuscitation fluids, at 20 DEG C, isothermal holding 15min, to obtain photobacterium phosphoreum resuscitation fluid；
S22 provides the osmotic pressure described in above-mentioned first embodiment and adjusts liquid, and osmotic pressure adjusting liquid is separately added into sample
In product solution and contrast solution, to obtain pretreatment sample and pretreatment control, and osmotic pressure adjusts liquid and sample solution and right
Mixed proportion according to solution is 1.0:9.0；
S23, is separately added into 10 μ L photobacterium phosphoreum resuscitation fluids into each pretreatment sample and pretreatment control, and with
20 DEG C of isothermal holding 15min of constant temperature, to obtain sample to be tested and control to be detected；
S24 is detected detected sample and control to be detected, to obtain the phase of detected sample and control to be detected
Close experimental data.
In first specific implementation mode of the present embodiment, the osmotic pressure provided in first embodiment of the invention is adjusted
In liquid, Mg2+Influence of the content to invention bacterium invention intensity is detected, and specific experimental group and contrast experiment are as follows：
Experimental group 11, provides and adjusts liquid just like the osmotic pressure described in first embodiment of the invention, and the NaCl contents are
20wt%, Hepes content are 15wt%, Mg2+Content is 1.0wt%, and the DMSO contents are 10% (V/V).By above-mentioned infiltration
Pressure adjusts liquid, and the method S20 that liquid carries out poisonous substance detection is adjusted using osmotic pressure according to a kind of described in third embodiment of the invention
It is detected；This experimental group only provides distilled water as sample simultaneously, other steps and parameter all with third embodiment of the invention
Identical in S20, details are not described herein.
Experimental group 12, difference lies in the osmotic pressure adjusts Mg described in liquid with above-mentioned experimental group 10 unique2+Content
For 2.0wt%.
Experimental group 13, difference lies in the osmotic pressure adjusts Mg described in liquid with above-mentioned experimental group 10 unique2+Content
For 3.0wt%.
Experimental group 14, difference lies in the osmotic pressure adjusts Mg described in liquid with above-mentioned experimental group 10 unique2+Content
For 4.0wt%.
Experimental group 15, difference lies in the osmotic pressure adjusts Mg described in liquid with above-mentioned experimental group 10 unique2+Content
For 5.0wt%.
Contrast groups 16, difference lies in the osmotic pressure adjusts Mg described in liquid with above-mentioned experimental group 10 unique2+Content
For 6.0wt%.
Control group 17, difference lies in the osmotic pressure adjusts Mg described in liquid with above-mentioned experimental group 10 unique2+Content
For 8.0wt%.
Control group 18, difference lies in the osmotic pressure adjusts Mg described in liquid with above-mentioned experimental group 10 unique2+Content
For 10.0wt%.
Control group 19, difference lies in the osmotic pressure adjusts Mg described in liquid with above-mentioned experimental group 10 unique2+Content
It is 0.
It in above-mentioned steps S24, is detected using water quality testing meter, the bright of above-mentioned experimental group and contrast groups is obtained with corresponding
Bright luminous bacillus luminous intensity.
Comparative analysis：Referring to Fig. 4, experimental group 11-15 is relative to control group 16-19, the bright hairs of the experimental group 11-15
The luminous intensity of polished rod bacterium is more than the luminous intensity of control group 16-19 photobacterium phosphoreums；The control group 16-18 luminous intensities
Less than the luminous intensity of control group 19.
In conclusion osmotic pressure adjusts Mg in liquid2+Content when not being more than 5.0wt%, photobacterium phosphoreum it is luminous strong
Degree is compared to being not added with Mg2+Luminous intensity, photobacterium phosphoreum luminous intensity maximum is enhanced to 7.2%, and in Mg2+Contain
It is big to the humidification of the luminous intensity of photobacterium phosphoreum when amount is 4.0%, the hair of photobacterium phosphoreum 7.2% can be enhanced
Luminous intensity；Work as Mg2+Content to be more than 5.0% be that the luminous intensity of photobacterium phosphoreum starts to be suppressed.
Its reason is：Include Mg2+Photobacteria in the necessary substance of luciferase reaction, therefore certain dense
It spends and increases Mg in range2+Content be conducive to improve and answer the activity of luciferase；And luciferase is that direct participation is luminous thin
The enzyme of bacterium luminescence-producing reaction, therefore increase Mg within the scope of a certain concentration2+Content be conducive to improve photobacteria luminous intensity.But
It is, as increased Mg2+Content it is excessive when, the luminescence-producing reaction of photobacteria can be inhibited, the luminous strong of photobacteria can be reduced
Degree.
In second specific implementation mode of the present embodiment, the osmotic pressure provided in first embodiment of the invention is adjusted
In liquid, DMSO contents are detected the luminous inhibiting rate of invention bacterium, and specific experimental group and contrast experiment are as follows：
Experimental group 21 provides the osmotic pressure described in a first embodiment of the invention and adjusts liquid, and the NaCl contents are
20wt%, Hepes content are 15wt%, Mg2+Content is 4.0wt%, and the DMSO contents are 5% (V/V).By above-mentioned osmotic pressure
Adjust liquid, according to described in third embodiment of the invention it is a kind of using osmotic pressure adjust liquid carry out poisonous substance detection method S20 into
Row experiment, and this experimental group only provides distilled water as sample, other steps and parameter all with third embodiment of the invention S20
In it is identical, details are not described herein.
Experimental group 22, difference lies in the osmotic pressure adjusts DMSO described in liquid with above-mentioned experimental group 10 unique
Content is 10% (V/V).
Experimental group 23, difference lies in the osmotic pressure adjusts DMSO described in liquid with above-mentioned experimental group 10 unique
Content is 15% (V/V).
Experimental group 24, difference lies in the osmotic pressure adjusts DMSO described in liquid with above-mentioned experimental group 10 unique
Content is 20% (V/V).
Control group 25, difference lies in the osmotic pressure adjusts DMSO described in liquid with above-mentioned experimental group 10 unique
Content is 25% (V/V).
Control group 26, difference lies in the osmotic pressure adjusts DMSO described in liquid and contains with above-mentioned experimental group 10 unique
Amount is 30% (V/V).
Control group 27, unique with above-mentioned experimental group 10 difference lies in the content of, the DMSO are 0.
It in above-mentioned steps S24, is detected using water quality testing meter, the bright of above-mentioned experimental group and contrast groups is obtained with corresponding
Bright luminous bacillus luminous intensity.
Comparative analysis：Referring to Fig. 5, experimental group 21-24 is relative to control group 25-27, the experimental group 21-24 and right
Can inhibition be generated to photobacterium phosphoreum luminous intensity, wherein experimental group 21-24 is to photobacterium phosphoreum according to group 25-27
Luminous intensity generates inhibition and is less than control group 25-27, and experimental group 21-22 is less than the inhibiting rate of photobacterium phosphoreum
5%.
Its reason is：DMSO itself belongs to organic species poisonous substance, only opposite with heavy metal, pesticide etc. for poisonous substances,
It is smaller to the toxicity of photobacteria, therefore the DMSO for adding low concentration can be smaller to the generation of photobacterium phosphoreum luminous intensity
Inhibiting effect.
The osmotic pressure provided in the present invention adjusts the method that liquid carries out poisonous substance detection, is promoted to poisonous substance detection sensitivity
Effect verified；Specific experiment step, according to described in third embodiment of the invention it is a kind of using osmotic pressure adjust liquid into
The method S20 of row poisonous substance detection is carried out, and specific experiment group is as follows：
(1) in the Hg containing different content2+In the sample solution of poisonous substance, using osmotic pressure adjust liquid and photobacteria into
Row poisonous substance detects, to obtain the luminous inhibiting rate of corresponding photobacteria.
Wherein, it is 15wt%, Mg that NaCl contents, which are 20wt%, Hepes content, in the osmotic pressure adjusting liquid2+Content is
4.0wt% and DMSO contents are 10% (V/V)；Successively prepare 0.02mg/L, 0.04mg/L, 0.06mg/L, 0.08mg/L,
The different Hg of 0.10mg/L and 0.12mg/L2+Toxic content sample solution, specific experimental group and contrast experiment are as follows：
Experimental group 31, to different Hg2+Osmotic pressure is added in toxic content sample and adjusts liquid, the osmotic pressure adjust liquid with it is upper
The mixed volume ratio for stating sample is 1:9.
Control group 32, difference lies in variant Hg with experimental group 312+30wt%NaCl is molten in toxic content sample
The mixed volume ratio of liquid, the 30wt%NaCl solution and above-mentioned sample is 1:9.
Comparative analysis：Referring to Fig. 6, experimental group 31, relative to control group 32, each sample is to bright in the experimental group 31
Luminous bacillus shines inhibiting rate, is better than photobacterium phosphoreum in control group 32 and shines inhibiting rate；Also, it shines carefully identical
Bacterium shines under inhibiting rate, Hg in experimental group 312+Toxic content concentration, both less than Hg in contrast groups 322+Toxic content concentration.
Its reason is：The osmotic pressure, which is adjusted, contains DMSO and Mg in liquid2+, it is thin that the DMSO can enhance photobacteria
The permeability of after birth promotes toxicant to be more easy to enter intracellular, Mg faster2+It a degree of can enhance photobacteria
Luminous intensity.
(2) containing different content 3, in the sample solutions of 5- Dichlorophenol poisonous substances, adjust liquid using osmotic pressure and shine
Bacterium carries out poisonous substance detection, to obtain the luminous inhibiting rate of corresponding photobacteria.
Wherein, it is 15wt%, Mg that NaCl contents, which are 20wt%, Hepes content, in the osmotic pressure adjusting liquid2+Content is
4.0wt% and DMSO contents are 10% (V/V).Successively prepare 3mg/L, 4mg/L, 5mg/L, 6mg/L, 7mg/L, 8mg/L and
The difference 3 of 9mg/L, 5- Dichlorophenol toxic content sample solutions, specific experimental group and contrast experiment are as follows：
Osmotic pressure adjusting liquid, the osmotic pressure tune are added into difference 3,5- Dichlorophenol toxic content samples for experimental group 41
The mixed volume ratio for saving liquid and above-mentioned sample is 1:9.
Control group 42, it is described with experimental group 41 difference lies in the addition 30wt%NaCl solution into each sample
The mixed volume ratio of 30wt%NaCl solution and above-mentioned sample is 1:9.
Comparative analysis：Referring to Fig. 7, experimental group 41, relative to control group 42, each sample is to bright in the experimental group 41
Luminous bacillus shines inhibiting rate, is better than photobacterium phosphoreum in control group 42 and shines inhibiting rate, especially in low content 3,5-
It is become apparent in the sample of Dichlorophenol toxic content concentration；Also, it shines under inhibiting rate in identical photobacteria, experimental group 41
In 3,5- Dichlorophenol toxic content concentration, both less than 3 in contrast groups 42,5- Dichlorophenol toxic content concentration.
Its reason is：Described 3,5- Dichlorophenols production asks down that solubility is low in water, and the osmotic pressure adjusts and contains in liquid
There are DMSO and Mg2+, the DMSO can effectively improve the solubility of 3,5- Dichlorophenols, while can also enhance light-emitting bacterial cells
The permeability of film promotes toxicant to be more easy to enter spirit that is intracellular, and then improving photobacteria pair 3,5- Dichlorophenols faster
Sensitivity；Mg simultaneously2+It can a degree of luminous intensity for enhancing photobacteria.
(3) in the testing sample solution of the decis poisonous substance containing different content, liquid and hair are adjusted using osmotic pressure
Photobacteria carries out poisonous substance detection, to obtain the luminous inhibiting rate of corresponding photobacteria.
Wherein, it is 15wt%, Mg that NaCl contents, which are 20wt%, Hepes content, in the osmotic pressure adjusting liquid2+Content is
4.0wt% and DMSO contents are 10% (V/V).1mg/L, 1.5mg/L, 2mg/L, 2.5mg/L, 3mg/L, 3.5mg/ are prepared successively
The different decis toxic content sample solutions of L and 4mg/L, specific experimental group and contrast experiment are as follows：
Osmotic pressure is added into different decis toxic content samples and adjusts liquid for experimental group 51, and the osmotic pressure is adjusted
The volume ratio of liquid and above-mentioned sample is 1:9.
Control group 52, it is described with experimental group 31 difference lies in the addition 30wt%NaCl solution into each sample
The volume ratio of 30wt%NaCl solution and above-mentioned sample is 1:9.
Comparative analysis：Referring to Fig. 8, experimental group 51, relative to control group 52, each sample is to bright in the experimental group 51
Luminous bacillus shines inhibiting rate, is better than photobacterium phosphoreum in control group 52 and shines inhibiting rate, especially in low concentration bromine cyanogen
It is become apparent in the sample of pyrethroids；Also, it shines under inhibiting rate in identical photobacteria, decis poisonous substance in experimental group 51
Content concn, decis toxic content concentration both less than in contrast groups 52.
Its reason is：Solubility is low in water for the decis, and the osmotic pressure is adjusted in liquid containing DMSO and
Mg2+, the DMSO can effectively improve the solubility of decis, while can also enhance the penetrating of light-emitting bacterial cells film
Property, promote toxicant to be more easy to faster enter into the cell, in turn, improves the sensitivity of toxicant；Mg simultaneously2+It can certain journey
The luminous intensity of the enhancing photobacteria of degree.
(4) in the sample solution of the nitrofen poisonous substance containing different content, liquid and photobacteria are adjusted using osmotic pressure
Poisonous substance detection is carried out, to obtain the luminous inhibiting rate of corresponding photobacteria.
Wherein, it is 15wt%, Mg that NaCl contents, which are 20wt%, Hepes content, in the osmotic pressure adjusting liquid2+Content is
4.0wt% and DMSO contents are 10% (V/V).0.5mg/L, 1mg/L, 1.5mg/L, 2mg/L, 2.5mg/L, 3mg/ are prepared successively
L, the different nitrofen toxic content sample solutions of 3.5mg/L and 4mg/L, specific experimental group and contrast experiment are as follows：
Experimental group 61, and osmotic pressure is added into different nitrofen toxic content samples and adjusts liquid, the osmotic pressure is adjusted
The volume ratio of liquid and above-mentioned sample is 1:9.
Control group 62, it is described with experimental group 61 difference lies in the addition 30wt%NaCl solution into each sample
The volume ratio of 30wt%NaCl solution and above-mentioned sample is 1:9.
Comparative analysis：Referring to Fig. 9, experimental group 61, relative to control group 62, each sample is to bright in the experimental group 61
Luminous bacillus shines inhibiting rate, is better than photobacterium phosphoreum in control group 62 and shines inhibiting rate, especially in low concentration weeding
It is become apparent in the sample of ether；Also, it shines under inhibiting rate in identical photobacteria, nitrofen toxic content in experimental group 61
Concentration, both less than or equal to nitrofen toxic content concentration in contrast groups 62.
Its reason is：Solubility is low in water for the nitrofen, and the osmotic pressure adjusts and contains DMSO and Mg in liquid2 +, the DMSO can effectively improve the solubility of nitrofen organic matter, while also enhance the permeability of light-emitting bacterial cells film,
Toxicant is promoted to be more easy to faster enter sensitivity that is intracellular, and then improving toxicant；Mg simultaneously2+It can be a degree of
Enhance the luminous intensity of photobacteria.
Compared with prior art, osmotic pressure provided by the present invention adjusts liquid, by adding magnesium compound, makes osmotic pressure tune
Mg is contained in section liquid2+, and by controlling Mg2+Its content effectively enhances the fluorescein for participating in photobacteria luminescence-producing reaction
The activity of enzyme, and then enhance the luminous intensity of photobacteria；Meanwhile the osmotic pressure adjusts liquid by adding Hepes and fitting
The NaOH solution of the 1mol/L of amount has adjusted the osmotic pressure and adjusts the pH value of liquid, and then the osmotic pressure is avoided to adjust liquid
PH value impacts sample P H values；Further, the osmotic pressure adjusts liquid, by being added to photobacteria hypotoxicity
DMSO solution can effectively increase the solubility of organic matter, it is particularly possible to improve insoluble in water, low solubility organic matter
And the solubility of low content organic matter, and DMSO solution can enhance the permeability of light-emitting bacterial cells film, make poisonous substance more
It is easily accessible inside photobacteria, and then improves photobacteria to insoluble organic matter, low-concentration organic, low dissolving in water
The detection sensitivity for spending the poisonous substances such as organic matter and other heavy metals makes osmotic pressure adjust liquid and promotes Luminous bacteria comprehensive in water quality
Close the application range of toxicity.
It is provided by the present invention that the method that liquid carries out poisonous substance detection is adjusted using osmotic pressure, by sample solution and control
A certain proportion of osmotic pressure is sequentially added in solution and adjusts liquid, recovery photobacteria solution, makes photobacteria when detecting sample,
With better luminous intensity and cell permeability, poisonous substance is set to be more easy to enter in photobacteria body faster, final photobacteria
Stronger luminous inhibiting rate is shown under identical concentration of poison, i.e. photobacteria only needs smaller concentration of poison, so that it may
To obtain identical luminous inhibiting rate, sensitivity of the photobacteria to poisonous substance is improved；Meanwhile by sample with compare in plus
Enter osmotic pressure and adjust liquid, makes poisonous substance, the low solubility organic matter of low concentration, especially improve the low solubilities organic matter such as pesticide
Solubility, further expand the detection sensitivity and application range of photobacteria poisonous substance detection method.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all originals in the present invention
Any modification made by within then, equivalent replacement and improvement etc. should all include within protection scope of the present invention.
Claims (10)
1. a kind of osmotic pressure adjusts liquid, it is characterised in that：Including NaCl, Hepes, magnesium compound, DMSO and suitable deionization
Water, wherein the NaCl contents are 20wt%-30wt%, the Hepes contents are 10wt%-20wt%, the magnesium compound
Content is 2.0wt%-5.0wt%, and the DMSO contents are 5%-20% (V/V).
2. osmotic pressure adjusts liquid as described in the appended claim 1, it is characterised in that：The NaCl contents are 20wt%, Hepes content
For 15wt%, magnesium compound content be 4.0wt% and DMSO contents it is 10% (V/V).
3. osmotic pressure adjusts liquid as described in the appended claim 1, it is characterised in that：It further includes that PH is adjusted that the osmotic pressure, which adjusts liquid,
Agent.
4. osmotic pressure adjusts liquid as claimed in claim 3, it is characterised in that：The PH conditioning agents are that the NaOH of 1mol/L is molten
Liquid.
5. osmotic pressure adjusts liquid as claimed in claim 4, it is characterised in that：The osmotic pressure adjusts the PH ranging from 7.4- of liquid
8.0。
6. osmotic pressure adjusts liquid as described in the appended claim 1, it is characterised in that：The magnesium compound includes MgCl2、MgSO4、Mg
(NO3)2Or Mg (OH)2One or more of.
7. a kind of adjusting the method that liquid carries out poisonous substance detection using osmotic pressure, it is characterised in that including：
Recovery photobacteria, to obtain recovery photobacteria bacterium solution；
The osmotic pressure as described in claim any one of 1-6 is provided and adjusts liquid, osmotic pressure is adjusted into liquid addition sample solution and right
According in solution, the osmotic pressure adjusts liquid and the sample solution mixed volume ratio is (1.5-0.5)：(8.5-9.5), it is described
Osmotic pressure adjusts liquid and adjusts liquid and the sample solution mixed volume ratio with the contrast solution mixed proportion and the osmotic pressure
Example is identical, to obtain pretreatment sample solution and pretreatment control solution；
Recovery photobacteria solution is added in pretreatment sample solution and pretreatment control solution, and every milliliter of sample solution or
The recovery photobacteria bacterium solution of 8-15 μ L is separately added into contrast solution, and isothermal holding obtains after a certain period of time at a certain temperature
Obtain sample to be tested and control to be checked；
The sample to be tested and control to be checked are detected, to obtain corresponding detection data.
8. as claimed in claim 7 adjust the method that liquid carries out poisonous substance detection using osmotic pressure, it is characterised in that：The infiltration
It is respectively 1 that pressure, which adjusts the volume ratio that liquid is separately added into the sample solution and contrast solution,：9.
9. as claimed in claim 7 adjust the method that liquid carries out poisonous substance detection using osmotic pressure, it is characterised in that：Every milliliter of sample
The recovery photobacteria bacterium solution of 10 μ L is separately added into product solution or contrast solution.
10. as claimed in claim 7 adjust the method that liquid carries out poisonous substance detection using osmotic pressure, it is characterised in that：The temperature
Degree is 20 DEG C, and the isothermal holding time is 15min or 30min.
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| CN102465167A (en) * | 2010-11-10 | 2012-05-23 | 中国科学院生态环境研究中心 | Rapid and high-flux acute toxicity test method for luminous bacteria |
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