CN103149199A - Manufacturing method of water-quality acute toxicity high throughput rapid determination reagent kit and using method thereof - Google Patents
Manufacturing method of water-quality acute toxicity high throughput rapid determination reagent kit and using method thereof Download PDFInfo
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Abstract
The invention relates to a manufacturing method of a water-quality acute toxicity high throughout rapid determination reagent kit and a using method of the water-quality toxicity high throughput rapid determine reagent kit. The manufacturing method of the toxicity high throughput rapid determination reagent kit comprises the following steps: (1) preparing photobacteria lyophilized powder; (2) configuring broth dilution water; (3) configuring phenol mother liquid; and (4) producing accessories and packing. The using method of the water-quality acute toxicity high throughout rapid determine reagent kit comprises the following steps: (1) reviving the bacteria lyophilized powder; (2) configuring standard solution of phenol series; (3) measuring residual chlorine; (4) measuring the potential hydrogen (pH) value; (5) preprocessing samples; (6) comparing and preparing dilution of the samples; (7) measuring accuracy of a standard curve of the phenol; (8) adding the photobacteria; (9) measuring light intensity; (10) calculating a result; (11) ensuring and controlling the quality; and (12) expressing the toxicity of the samples. The manufacturing method of the water-quality acute toxicity high throughput rapid determination reagent kit and the using method of the water-quality acute toxicity high throughput rapid determine reagent kit have the advantages of being applicable to rapid measurement of the water-quality toxicity, perfect in manufacturing and using, low in cost, and rapid and accurate in measurement.
Description
Technical field
The present invention relates to a kind of technical field of kit, specifically a kind of method for making and using method of utilizing the kit that the fresh water photobacteria measures water quality material acute toxicity.
Background technology
At present; the method that adopts fresh water Qinghai Vibrion assessment water quality toxicity is provided in " water and effluent monitoring analytical approach " (the 4th edition); before requiring test, employing standard toxicant phenol (solvent is 10% lactose solution) carries out quality testing to photobacteria; under good state, water quality toxicity is made accurate response with the assurance photobacteria, and sample to be checked also must add 10% lactose.There is following problem in the method in process of the test: the first, 10% lactose solution is unstable, can only preserve 2d with the phenol mother liquor of 10% lactose configuration at most in the refrigerator of 4 ℃, should measure immediately after configuration; And the 1000mg/L mother liquor of 100ml and the series standard solution of 450ml only use respectively 63.5ml and 18ml in experimentation, so just make phenol solution not to be fully used, caused very large waste, collected phenol waste liquid has increased cost and difficulty that objectionable impurities is disposed, may cause serious pollution to environment.The second, 10% lactose is difficult to the nature dissolving after adding sample, needs heating in course of dissolution, can change like this character of sample, affects the accuracy of result.The 3rd, because the luminous intensity of photobacteria can change along with the variation of time, so adopt conventional photometer to carry out single light intensity when detecting, measuring a sample just needs 1 contrast of mensuration, both wastes time and energy like this, increases again cost.For warranty test is completed quickly and accurately, need to improve existing method, the making and use method of fresh water photobacteria to the kit of water quality material acute toxic test is provided simultaneously.
Summary of the invention
The objective of the invention is in order to solve the problems of the technologies described above, proposing a kind of acute toxicity of water quality high flux quick measuring reagent box makes and using method, existing photogen freeze-dried powder and freeze-dried powder resuscitation fluid thereof in this kit are accompanied with again the serial phenol solution for evaluation photobacteria quality.This kit can be measured as required at any time, also can be used for open-air water sample emergency monitoring, greatly simplify the program of former test, has saved the time, has economy, advantage fast and efficiently.
The objective of the invention is to be achieved through the following technical solutions:
Acute toxicity of water quality high flux quick measuring reagent box method for making of the present invention is characterized in that having following steps:
1) preparation of photogen freeze-dried powder:
(a) cultivation of bacterium: select the photobacteria of cultivating under laboratory condition, cultivate exponential phase, can observe glaucous fluorescence in the darkroom;
(b) bacterium liquid is concentrated: at 4 ℃, under the condition of 5000r/min, refrigerated centrifuge 15min, abandon supernatant with the bacterium liquid of exponential phase;
(c) bacterium liquid suspends: in the skim milk of the bacterium liquid Eddy diffusion after concentrating in 10%, 5% lactose and 0.8% the mixed uniformly preservative agent of sodium chloride;
(d) packing: use pipettor to draw 2ml photogen suspending liquid from above-mentioned bacterium liquid to the bottle that can seal;
(e) freezing and preservation: at-70 ℃ of pre-freeze 3h, vacuum freezedrying 24～48h is placed on preservation in-20 ℃ of refrigerators the air-tight bottle that bacterium suspending liquid is housed.
2) configuration of bacterium liquid dilution water:
(a) accurately take 0.8g sodium chloride, add the distilled water dissolving of 80ml;
(b) be transferred in the volumetric flask of 100ml after the dissolving, add distilled water to the graticule of 100ml, shake up;
(c) do not contain any known material poisonous to Daphnia magna in the inspection dilution water: any in chlorine, heavy metal, agricultural chemicals, ammonia, polychlorinated biphenyl;
(d) be transferred in aseptic pp bottle 4 ℃ of preservations after autoclaving;
3) configuration of phenol mother liquor:
(a) get phenol crystal and heat in cucurbit, collect the cut of 181 ℃～183 ℃ with small beaker;
(b) when the cut temperature reduces the wish condensation, constantly stir until be colourless loose sand shape with glass bar;
(c) pack into fast in brown bottle, after good seal, low-temperature dark is preserved;
(d) accurately take rapidly 0.0100g phenol, add the chromatographically pure distilled water dissolving of 3ml, then be transferred in the brown volumetric flask of 5ml, add entry to graticule, shake up;
(e) afterwards mother liquor is transferred to rapidly in the 5ml sample bottle that can seal, is labeled as 0 outside bottle, after sealing, 4 ℃ keep in Dark Place.
4) make accessory and mounted box:
(a) preparation of phenol series standard solution bottle: prepare the vial of 9 5ml, take the lactose of 0.25g in each bottle, indicate respectively 1. above bottle, 2., 3., 4., 5., 6., 7., 8., 9., 4 ℃ of sealings are preserved;
(b) prepare the sampling jar of 8 50ml, the lactose of the 2.5g that packs in each bottle, 4 ℃ of sealings are preserved;
(c) prepare the pH test paper;
(d) prepare sodium thiosulfate: take sodium thiosulfate, specification is the 70mg/ bag, prepares 7 bags.
(e) be marked with 96 orifice plates of blank and experimental group printed words, and with 2 of the sealed membranes that conforms to 96 orifice plate specifications;
Afterwards the bacterium freeze-dried powder for preparing in above-mentioned steps, bacterium liquid dilution water, phenol mother liquor and accessory are packed in box.
The using method of kit of the present invention is characterized in that following steps are arranged:
1) recovery of bacterium freeze-dried powder: freeze-dried powder and resuscitation fluid are placed 15min in the refrigerator of 4 ℃, then resuscitation fluid is injected the bottle that the photogen freeze-dried powder is housed fast, afterwards this bacterium liquid is transferred in a clean tube, then per sample amount adds appropriate resuscitation fluid, the standing 10～15min that stablizes after mixing;
2) configuration of phenol series standard solution: be configured to 20,40,80,100,120,160,200,250 and the serial phenol solution of 300mg/L with the phenol mother liquor, collocation method be indicate 1., 2., 3., 4., 5., 6., 7., 8., add respectively 4.9,4.8,4.6,4.5,4.4,4.2,4.0,3.75 and the distilled water of 3.5ml in 9. bottle, the phenol mother liquor that adds respectively 100,200,400,500,600,800,1000,1250 and 1500 μ l after dissolving, add rear sealing, mixing is to be measured;
3) measure chlorine residue: if sample does not contain chlorine residue, do not need to add sodium thiosulfate, if chlorinty greater than detection limit, adds sodium thiosulfate, until do not contain chlorine residue in sample;
4) measure the pH value: measure the pH value of contrast and experimental group, the pH value can be carried out Sample Dilution and be prepared in 6.0～8.5 scope; If sample does not meet the demands, need to use the NaOH of 1mol/L or the HCl of 1mol/L to carry out the adjustment of pH value;
5) determination: add 50ml distilled water and sample in the sample bottle that the 2.5g lactose is housed, distinguish as a control group and experimental group mixing;
6) contrast and Sample Dilution are prepared: with 5% lactose solution in contrast, and water sample dilutes according to following principle, if the larger specimen test concentration of toxicity is pressed the concentration series of geometric series, i.e. and 5-7 concentration of geometric progression spacing design; Geometric ratio level coefficient is no more than 2.2, be 2 as 0.5,1,2,4,8,16 geometric ratio level coefficients, ideal in 40%～60% concentration inhibiting rate can occur in series concentration, and toxicity is less, the relative inhibition of photogen is not needed to arrange series concentration less than 50% sample, only need to carry out the mensuration of 100% concentration; The contrast with each concentration of sample all establish two parallel, add respectively in the hole of 96 orifice plates, each aerial addition is 200 μ l;
7) measure the degree of accuracy of phenol Standard curve: when each working sample, all will make the phenol Standard curve with identical bacterium liquid, get respectively the phenol solution 200 μ l of each concentration to each hole;
8) add photogen: add the bacteria suspension of 20 μ l in each hole, cover sealed membrane on 96 hole test boards, begin timing after mixing;
9) measure luminous intensity: after sample and photobacteria accurately act on 15min and 30min, with the luminous intensity of 96 micropore template multifunctional light degree meters at 450nm place mensuration contrast, phenol and sample;
10) result is calculated:
The relative light rate that presses down according to following formula calculation sample and phenol
11) quality assurance and quality control
A) accuracy of phenol Standard curve: calculate according to monobasic once linear regression equation, be that phenol should meet monobasic once linear regression equation I=a+bC to relatively pressing down between light rate (I) and phenol concentration (C) of photogen, the concentration (EC of phenol when suppressing 50% luminosity factor
50) should be between 100～200mg/L;
B) precision of method: the relative error of twice replicate determination result of sample is less than or equal to 10%.
12) expression of sample toxicity
A) calculate EC
50: when suppressing 50% luminosity, the dilution percentage concentration of sample is EC
50, be that the Kou Shi revised law obtains EC with computer program
50
B) calculate toxic unit TU:
According to formula TU=1/EC
50Calculate TU, sample toxicity is sorted out according to the size of TU value, and TU=0 is that nontoxic, 0＜TU＜1 is little poison, and 1＜TU＜10 be poisonous, and 10＜TU＜100 are for extremely malicious, and TU＞100 are hypertoxic.
Beneficial effect of the present invention: greatly reduced the use amount of phenol and the generation of phenol waste liquid in the mensuration process of positive control phenol series concentration, cost and difficulty that phenol is disposed have been reduced, effectively protected environment to avoid secondary pollution, the probability that simultaneously in whole process, human body is exposed to phenol reduces greatly, can effectively avoid testing crew to be subject to the pollution of phenol; This kit is ready to lactose in advance, and this lactose concn 5% is obviously different from 10% concentration, 5% lactose can be dissolved in sample at normal temperatures, and 10% lactose must heat and long-time stirring, the improvement of lactose concn has improved accuracy and the reliability of measurement result, makes this kit can be used for open-air emergency monitoring; This kit has prepared photogen freeze-dried powder, resuscitation fluid, phenol mother liquor, 96 orifice plates, the lactose that can directly use in advance, only measures to add water sample, has therefore saved manpower and the financial cost of bacterium long-term cultivation; A kit can obtain 96 data simultaneously, not only can avoid manual operation error and instrument restriction, reduce simultaneously the detection error that difference causes in incubation due to bacterium, can well reduce the detection error between the laboratory.
Embodiment
The present invention is described further below in conjunction with embodiment.
Embodiment 1: acute toxicity of water quality high flux quick measuring reagent box method for making, make according to the following steps:
1) preparation of photogen freeze-dried powder:
(a) cultivation of bacterium: select the photobacteria of cultivating under laboratory condition, cultivate exponential phase, can observe glaucous fluorescence in the darkroom;
(b) bacterium liquid is concentrated: at 4 ℃, under the condition of 5000r/min, refrigerated centrifuge 15min, abandon supernatant with the bacterium liquid of exponential phase;
(c) bacterium liquid suspends: in the skim milk of the bacterium liquid Eddy diffusion after concentrating in 10%, 5% lactose and 0.8% sodium chloride preservative agent;
(d) packing: use pipettor to draw 2ml photogen suspending liquid from above-mentioned bacterium liquid to the bottle that can seal;
(e) freezing and preservation: at-70 ℃ of pre-freeze 3h, vacuum freezedrying 24～48h is placed on preservation in-20 ℃ of refrigerators the air-tight bottle that bacterium suspending liquid is housed.
2) configuration of bacterium liquid dilution water:
(a) accurately take 0.8g sodium chloride, add the distilled water dissolving of 80ml;
(b) be transferred in the volumetric flask of 100ml after the dissolving, add entry to the graticule of 100ml, shake up;
(c) do not contain any known material poisonous to Daphnia magna in the inspection dilution water: any in chlorine, heavy metal, agricultural chemicals, ammonia, polychlorinated biphenyl;
(d) be transferred in aseptic pp bottle 4 ℃ of preservations after autoclaving;
3) configuration of phenol mother liquor:
(a) get phenol crystal and heat in cucurbit, collect the cut of 182 ℃ with small beaker;
(b) when the cut temperature reduces the wish condensation, constantly stir until be colourless loose sand shape with glass bar;
(c) pack into fast in brown bottle, after good seal, low-temperature dark is preserved;
(d) accurately take rapidly 0.0100g phenol, add appropriate chromatographically pure distilled water dissolving, then be transferred in the brown volumetric flask of 5ml, add entry to graticule, shake up;
(e) afterwards mother liquor is transferred to rapidly in the 5ml sample bottle that can seal, is labeled as 0 outside bottle, after sealing, 4 ℃ keep in Dark Place.
4) make accessory and mounted box:
(a) preparation of phenol series standard solution bottle: prepare the vial of 9 5ml, take the lactose of 0.25g in each bottle, indicate respectively 1. above bottle, 2., 3., 4., 5., 6., 7., 8., 9., 4 ℃ of sealings are preserved;
(b) prepare the sampling jar of 8 50ml, take the lactose of 2.5g in each bottle, 4 ℃ of sealings are preserved;
(c) prepare the pH test paper;
(d) sodium thiosulfate: prepare sodium thiosulfate, the 70mg/ bag is prepared 7 bags;
(e) be marked with 96 orifice plates of blank and experimental group printed words, and with 2 of the sealed membranes that conforms to 96 orifice plate specifications;
Afterwards the bacterium freeze-dried powder for preparing in above-mentioned steps, bacterium liquid dilution water, phenol mother liquor and accessory are packed in box.
Embodiment 2: acute toxicity of water quality high flux quick measuring reagent box using method, and the sample of mensuration is total water inlet of sewage treatment plant, carries out according to the following steps:
1) recovery of bacterium freeze-dried powder: freeze-dried powder and resuscitation fluid are placed 15min in the refrigerator of 4 ℃, then resuscitation fluid is injected the bottle that the photogen freeze-dried powder is housed fast, afterwards this bacterium liquid is transferred in a clean tube, then per sample amount adds appropriate resuscitation fluid, the standing 10～15min that stablizes after mixing;
2) configuration of phenol series standard solution: be configured to 20,40,80,100,120,160,200,250 and the serial phenol solution of 300mg/L with the phenol mother liquor, collocation method be indicate 1., 2., 3., 4., 5., 6., 7., 8., add respectively 4.9,4.8,4.6,4.5,4.4,4.2,4.0,3.75 and the distilled water of 3.5ml in 9. bottle, the phenol mother liquor that adds respectively 100,200,400,500,600,800,1000,1250 and 1500 μ l after dissolving, add rear sealing, mixing is to be measured;
3) measure chlorine residue: total chlorine of sample does not need to add sodium thiosulfate lower than detection limit;
4) measure the pH value: the pH value of mensuration contrast and total water inlet is respectively 7.61 and 7.84, satisfies test request, does not need to add adjusting pH value;
5) determination: add respectively 50ml distilled water and sample in 2 sample bottles of 2.5g lactose are housed, distinguish as a control group and experimental group mixing;
6) contrast and Sample Dilution are prepared: with 5% lactose solution in contrast, and the experimental concentration of total water inlet is set to respectively 3.125%, 6.25%, 12.5%, 25%, 50% and 100%, and geometric ratio level coefficient is 2; The contrast with each concentration of sample all establish two parallel, add respectively in the hole of 96 orifice plates, each aerial addition is 200 μ l;
7) measure the degree of accuracy of phenol Standard curve: when each working sample, all will make the phenol Standard curve with identical bacterium liquid, get respectively the phenol solution 200 μ l of each concentration to each hole;
8) add photogen: add the bacteria suspension of 20 μ l in each hole, cover sealed membrane on 96 hole test boards, begin timing after mixing;
9) measure luminous intensity: sample and photobacteria accurately act on the luminosity of measuring contrast, phenol and sample after 15min and 30min, and measurement result is:
10) result is calculated:
The relative light rate that presses down according to following formula calculation sample and phenol
At first calculate the mean value of contrast luminosity=(121356+121657+121093)/3, trying to achieve mean value is 121369,
Then according to top formula
Calculate the relative of phenol and sample
Press down light rate (seeing the following form):
11) quality assurance and quality control
A) accuracy of phenol Standard curve
According to monobasic once linear regression equation Pyrogentisinic Acid, relatively pressing down between light rate (I) and phenol concentration (C) of photogen returned, obtain I=0.304C+1.366, R
2=0.99, the concentration EC of phenol when suppressing 50% luminosity factor
50Be 160mg/L;
B) precision of method
The relative error of twice replicate determination result of sample is less than or equal to 10%;
12) expression of sample toxicity
A) calculate EC
50
When suppressing 50% luminous quantity, the dilution percentage concentration of this sample is EC
50, be that the Kou Shi revised law is processed acquisition EC with computer program
50
Calculate 3.125%, 6.25%, 12.5%, 25%, 50% and be respectively 9%, 13%, 22%, 34%, 59% and 91% with the relative mean value that presses down the light rate of 100% sample, the concentration of sample meets I=84.96C+10.09, R with the relative light rate that presses down
2=0.98 linear dependence, the sample concentration EC when calculating I=50% according to I=84.96C+10.09
50, C=(50-10.09)/84.96) and be 46.98%;
B) calculate toxic unit TU:
According to formula TU=1/EC
50Calculate TU, TU=1/46.98%, calculating TU is 2.13, sorts out according to the size of TU value and determines total water inlet for poisonous, namely 1＜TU＜10 are poisonous.
The present embodiment has been made 24, has used 24 times in reality test, compares with the kit of routine, and the cost of manufacture of each box has reduced by 80%, and fabrication cycle is short 50%; The result of actual test shows that the precision of the accuracy of phenol Standard curve and method in acceptable scope, has reached purpose quick, accurate, reliable, simple to operate.
Claims (2)
1. acute toxicity of water quality high flux quick measuring reagent box method for making is characterized in that having following steps:
1) preparation of photogen freeze-dried powder:
(a) cultivation of bacterium: select the photobacteria of cultivating under laboratory condition, cultivate exponential phase, can observe glaucous fluorescence in the darkroom;
(b) bacterium liquid is concentrated: at 4 ℃, under the condition of 5000r/min, refrigerated centrifuge 15min, abandon supernatant with the bacterium liquid of exponential phase;
(c) bacterium liquid suspends: in the skim milk of the bacterium liquid Eddy diffusion after concentrating in 10%, 5% lactose and 0.8% the mixed uniformly preservative agent of sodium chloride;
(d) packing: use pipettor to draw 2ml photogen suspending liquid from above-mentioned bacterium liquid to the bottle that can seal;
(e) freezing and preservation: at-70 ℃ of pre-freeze 3h, vacuum freezedrying 24～48h is placed on preservation in-20 ℃ of refrigerators the air-tight bottle that bacterium suspending liquid is housed.
2) configuration of bacterium liquid dilution water:
(a) accurately take 0.8g sodium chloride, add the distilled water dissolving of 80ml;
(b) be transferred in the volumetric flask of 100ml after the dissolving, add distilled water to the graticule of 100ml, shake up;
(c) do not contain any known material poisonous to Daphnia magna in the inspection dilution water: any in chlorine, heavy metal, agricultural chemicals, ammonia, polychlorinated biphenyl;
(d) be transferred in aseptic pp bottle 4 ℃ of preservations after autoclaving;
3) configuration of phenol mother liquor:
(a) get phenol crystal and heat in cucurbit, collect the cut of 181 ℃～183 ℃ with small beaker;
(b) when the cut temperature reduces the wish condensation, constantly stir until be colourless loose sand shape with glass bar;
(c) pack into fast in brown bottle, after good seal, low-temperature dark is preserved;
(d) accurately take rapidly 0.0100g phenol, add the chromatographically pure distilled water dissolving of 3ml, then be transferred in the brown volumetric flask of 5ml, add entry to graticule, shake up;
(e) afterwards mother liquor is transferred to rapidly in the 5ml sample bottle that can seal, is labeled as 0 outside bottle, after sealing, 4 ℃ keep in Dark Place.
4) make accessory and mounted box:
(a) preparation of phenol series standard solution bottle: prepare the vial of 9 5ml, take the lactose of 0.25g in each bottle, indicate respectively 1. above bottle, 2., 3., 4., 5., 6., 7., 8., 9., 4 ℃ of sealings are preserved;
(b) prepare the sampling jar of 8 50ml, the lactose of the 2.5g that packs in each bottle, 4 ℃ of sealings are preserved;
(c) prepare the pH test paper;
(d) prepare sodium thiosulfate: take sodium thiosulfate, specification is the 70mg/ bag, prepares 7 bags.
(e) be marked with 96 orifice plates of blank and experimental group printed words, and with 2 of the sealed membranes that conforms to 96 orifice plate specifications;
Afterwards the bacterium freeze-dried powder for preparing in above-mentioned steps, bacterium liquid dilution water, phenol mother liquor and accessory are packed in box.
2. the using method of an acute toxicity of water quality high flux quick measuring reagent box is characterized in that following steps are arranged:
1) recovery of bacterium freeze-dried powder: freeze-dried powder and resuscitation fluid are placed 15min in the refrigerator of 4 ℃, then resuscitation fluid is injected the bottle that the photogen freeze-dried powder is housed fast, afterwards this bacterium liquid is transferred in a clean tube, then per sample amount adds appropriate resuscitation fluid, the standing 10～15min that stablizes after mixing;
2) configuration of phenol series standard solution: be configured to 20,40,80,100,120,160,200,250 and the serial phenol solution of 300mg/L with the phenol mother liquor, collocation method be indicate 1., 2., 3., 4., 5., 6., 7., 8., add respectively 4.9,4.8,4.6,4.5,4.4,4.2,4.0,3.75 and the distilled water of 3.5ml in 9. bottle, the phenol mother liquor that adds respectively 100,200,400,500,600,800,1000,1250 and 1500 μ l after dissolving, add rear sealing, mixing is to be measured;
3) measure chlorine residue: if sample does not contain chlorine residue, do not need to add sodium thiosulfate, if chlorinty greater than detection limit, adds sodium thiosulfate, until do not contain chlorine residue in sample;
4) measure the pH value: measure the pH value of contrast and experimental group, the pH value can be carried out Sample Dilution and be prepared in 6.0～8.5 scope; If sample does not meet the demands, need to use the NaOH of 1mol/L or the HCl of 1mol/L to carry out the adjustment of pH value;
5) determination: add 50ml distilled water and sample in the sample bottle that the 2.5g lactose is housed, distinguish as a control group and experimental group mixing;
6) contrast and Sample Dilution are prepared: with 5% lactose solution in contrast, and water sample dilutes according to following principle,
If the specimen test concentration that toxicity is larger is pressed the concentration series of geometric series, i.e. 5-7 concentration of geometric progression spacing design; Geometric ratio level coefficient is no more than 2.2, be 2 as 0.5,1,2,4,8,16 geometric ratio level coefficients, ideal in 40%～60% concentration inhibiting rate can occur in series concentration, and toxicity is less, the relative inhibition of photogen is not needed to arrange series concentration less than 50% sample, only need to carry out the mensuration of 100% concentration; The contrast with each concentration of sample all establish two parallel, add respectively in the hole of 96 orifice plates, each aerial addition is 200 μ l;
7) measure the degree of accuracy of phenol Standard curve: when each working sample, all will make the phenol Standard curve with identical bacterium liquid, get respectively the phenol solution 200 μ l of each concentration to each hole;
8) add photogen: add the bacteria suspension of 20 μ l in each hole, cover sealed membrane on 96 hole test boards, begin timing after mixing;
9) measure luminous intensity: after sample and photobacteria accurately act on 15min and 30min, with the luminous intensity of 96 micropore template multifunctional light degree meters at 450nm place mensuration contrast, phenol and sample;
10) result is calculated:
The relative light rate that presses down according to following formula calculation sample and phenol
11) quality assurance and quality control
A) accuracy of phenol Standard curve: calculate according to monobasic once linear regression equation, be that phenol should meet monobasic once linear regression equation I=a+bC to relatively pressing down between light rate (I) and phenol concentration (C) of photogen, the concentration EC of phenol when suppressing 50% luminosity factor
50, should be between 100～200mg/L;
B) precision of method: the relative error of twice replicate determination result of sample is less than or equal to 10%.
12) expression of sample toxicity
A) calculate EC
50: when suppressing 50% luminosity, the dilution percentage concentration of sample is EC
50, be that the Kou Shi revised law obtains EC with computer program
50
B) calculate toxic unit TU:
According to formula TU=1/EC
50Calculate TU, sample toxicity is sorted out according to the size of TU value, and TU=0 is that nontoxic, 0＜TU＜1 is little poison, and 1＜TU＜10 be poisonous, and 10＜TU＜100 are for extremely malicious, and TU＞100 are hypertoxic.
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| CN104419644A (en) * | 2013-09-10 | 2015-03-18 | 上海市刑事科学技术研究院 | Luminous bacteria freeze-dried preparation and preparation method thereof |
| CN105973875A (en) * | 2016-04-27 | 2016-09-28 | 四川省中医药科学院 | Quality control method for drug microtox assay system |
| CN106596525A (en) * | 2016-12-30 | 2017-04-26 | 深圳市世纪天源环保技术有限公司 | Water biological toxicity detection method and device |
| CN106771029A (en) * | 2016-12-16 | 2017-05-31 | 常州市环境监测中心 | One kind is used for Daphnia magna acute toxicity detection kit |
| CN110441223A (en) * | 2019-07-11 | 2019-11-12 | 南京信息职业技术学院 | A method of solid particle toxicity evaluation is carried out using photobacteria |
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