CN107632164A - A kind of inexpensive Ag+Detection method - Google Patents
A kind of inexpensive Ag+Detection method Download PDFInfo
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- CN107632164A CN107632164A CN201710840168.7A CN201710840168A CN107632164A CN 107632164 A CN107632164 A CN 107632164A CN 201710840168 A CN201710840168 A CN 201710840168A CN 107632164 A CN107632164 A CN 107632164A
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Abstract
The invention discloses a kind of inexpensive Ag+Detection method, finite concentration, the glucose oxidase solution and Ag+ solution of certain volume are added first in test tubes, allow its reaction a period of time at a certain temperature；Then, the glucose solution of certain volume is added, allows it to react the regular hour；Then, the IodineSodium Solution of concentration known is added, allows it to react certain time；Finally, certain density soluble starch solution is added, observes the change of solution colour；Meanwhile the solution of generation elemental iodine is added dropwise in the sample application zone of paper microfluidic analysis device.Finally, by observing the change can of solution colour in ELISA Plate qualitatively Ag in analytical solution+Content and measurement paper microfluidic analysis device detection zone generation blue trace length to Ag+Carry out quantitative analysis.The present invention does not need the operation of instrument and equipment, and cost is cheap, simple to operate.
Description
Technical field
The present invention relates to Ag+A kind of detection field, and in particular to inexpensive Ag+Detection method.
Background technology
Many heavy metal ion are dispersed among the environment of people's life, it can enter people's in several ways
Among life.Such as：Among the daily diet of people；During workpeople's exploit mineral resources；The production of industrial products
During；Among untreated sewage, sludge.The pollution of heavy metal is formed to the health of people and the stable of the ecosystem
Increasingly severe threat.Heavy metal ion can conform to the part in the group containing sulfydryl related to life entity and correlation
On, so as to inhibit the metabolic process of glutathione and the activity of substantial amounts of enzyme and hormone.Heavy metal ion is necessary with life entity
Trace element be different, they often substitute the necessary trace element of life entity to be incorporated in protein, acceptor, metalloenzyme
On, so as to result in the serious diseases of human metabolism's process.Be engaged in it is biomedical with the people of environmental science to it is various not
Same heavy metal ion suppresses have very big interest in the activity of various different enzymes, for example researchers' heavy metal ion exists
Combined amount in unit enzyme molecule and its suppression percentage to enzyme molecule activity are studied and learnt in detail.Its
In, Pb2+、Hg2+、Cd2+、Cu2+And Ag+This five heavy metal species ion is often had an effect with enzyme, so as to cause enzyme to inactivate or live
Property reduce.Researchers have studied Hg2+And Cu2+To the inhibitory action of many enzymes.But for Ag+With the phase interaction of enzyme
Research is very few.Ag+It is one kind in heavy metal ion.Meanwhile it be also it is a kind of most virose, can be with carcinogenic dirt
Contaminate thing.It is typically found in a variety of metallic ores.Industrially, silver is usually utilized to make silver nitrate, silver bromide
And other chemicals relevant with photography.In life, silver is present on day conventional mirror, on silverer, special battery
Electrode on, in the tableware on dining table, on the jewellery of daily wearing, among dental care and other scientific equipments.At some
Country, people are carried out disinfection processing with silver oxide to daily drinking water, so will result in concentration of silver ions liter in running water
It is high.However, during people largely use silver nano material and nano-particle, silver ion will enter aquatic ecosystem
In system, just to environment, there may be serious harm for this.People are discharged into the silver ion in water body, soil, and it can be enriched with
Necessary to daily life among food, such as：The marine product such as various fish and shell, so as to which the health to people causes
Titanic peril.Linen spot caused by skin and the other organ-tissues of human body is that silver ion poisoning is most obvious special
Sign.US Gov Env Protection Agency set up the content standard of heavy metal ion in food and drinking-water.Wherein, Ag+In drinking water
In concentration can not be more than 0.46 μM.Because heavy metal ion is to the toxicity of life entity, therefore people are highly desirable to heavy metal
Ion is measured and micro heavy ion is detected.Nowadays exist in a variety of method detection ecological environments, life
Heavy metal ion in thing science, in food samples.These methods include：Atomic absorption spectroscopy, inductive etc. from
Sub- emission spectrum and Inductively coupled plasma-mass spectrometry, atomic fluorescence spectrometry, x-ray fluorescence spectrometry method and electrochemical analysis
Method.However, these methods are required for complicated, accurate instrument and equipment to carry out analysis detection, and gone back for the operator of instrument
Need to carry out the training and study of special instrument application method.Therefore, people especially need a kind of low cost of design and development,
Quick detection, the analysis method of high sensitivity are to Ag+Carry out analysis detection.
The content of the invention
To solve the above problems, the invention provides a kind of inexpensive Ag+Detection method, it is not necessary to the behaviour of instrument and equipment
Make, cost is cheap, simple to operate, can realize Ag+Quick detection, high sensitivity.
To achieve the above object, the technical scheme taken of the present invention is：
A kind of inexpensive Ag+Detection method, comprise the following steps：
S1, first on computers using the figure needed for Corel DRAW Software on Drawing, wherein, the diameter of circular sample application zone
For 5.0mm, the width and length in hough transform area are respectively that 3.0mm and 40.0mm, detection zone come with interval 3mm scale；
S2, after the completion for the treatment of graphic plotting, a 9cm × 9cm quantitative Medium speed filter paper is positioned over to the work of laser engraving machine
Make on platform, start instrument, by laser engraving machine, required paper microfluidic analysis device is cut out on filter paper, it is micro-fluidic in paper
The detection zone of analytical equipment, it is molten that 2% soluble starch configured by formamide solution is uniformly coated with 1 μ L microsyringe
The μ L of liquid 1, dry under field conditions (factors), be stored in the clean place without dust, it is standby；
S3, in 1.5mL centrifuge tube, sequentially add the μ g/mL of certain volume 2 GOD solution and certain density Ag+It is molten
Liquid, then it is placed in 40 DEG C of constant water bath box, after reacting 15min, 20mM glucose solution is added into centrifuge tube,
Continue to react 30min；Then 0.5M IodineSodium Solution is added into centrifuge tube, continues to react 5min；
After the completion of S4, reaction, centrifuge tube is taken out from constant water bath box, is then added dropwise with liquid-transfering gun draw solution in step
The sample application zone of paper microfluidic analysis device obtained by S2, analytical solution flow into paper microfluidic analysis device under capillarity
Detection zone, the 2% soluble starch solution that detection zone is fixed run into the elemental iodine in analytical solution, produce coloured rail
Mark, completed by the length for measuring the coloured track of detection zone to Ag+Quantitative detection.
The invention has the advantages that：
It is simple to operate, exempt from instrument, cost is cheap, and phenomenon is obvious, and detection limit is low, can detect 100fM Ag+Concentration.
New method is suitable for promoting the use of in the fields such as the limited remote backward areas water quality of economic condition, food, environment measuring.
Brief description of the drawings
Fig. 1 is the schematic diagram of the embodiment of the present invention.
Feasibility Experiments of the Fig. 2 based on paper microfluidic analysis device detection silver ion.
Fig. 3 is response of the length of paper microfluidic analysis device detection zone blueness trace to concentration of silver ions.(a. is high, in,
Response of low three concentration of silver ions to detection zone blueness contact length；B. the linear detection range of silver ion；Each data are 6
The average value of secondary parallel laboratory test, gained standard deviation is as error bar.)
Fig. 4 is based on paper microfluidic analysis device detection Ag+Specificity experiments.(wherein, 1.Ag+、2.Cd2+、3.Co2+、
4.Cr2+、5.Cu2+、6.Fe2+、7.Hg2+、8.Mn2+、9.Ni2+、10.Pb2+、11.Zn2+。)
Embodiment
In order that objects and advantages of the present invention are more clearly understood, the present invention is carried out with reference to embodiments further
Describe in detail.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to limit this hair
It is bright.
Reagent needed for following examples is shown in Tables 1 and 2 respectively with instrument.
The experiment reagent of table 1
PBS cushioning liquid (0.1M, pH 6.0) is configured by disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium chloride and potassium chloride；
2% soluble starch solution is configured by formamide solution；Glucose solution, glucose oxidase (GOD) solution are by PBS solution
Configuration.Unless stated otherwise, otherwise all solution are configured and diluted using deionized water (resistivity is more than 18M Ω .cm).
The laboratory apparatus of table 2
Used 2% soluble starch solution is as obtained by being prepared following steps：
First, the small beaker (being soaked in advance with chloroazotic acid) that two capacity are 25mL is taken, numbering is respectively A and B.Then, in A
3mL formamide solution is added in beaker；6.5mL formamide solutions are added in B beakers.Then, in fume hood, B beakers are given
In formamide solution put foam circle and be placed in constant temperature water tank and heated, formamide solution is heated to 100 DEG C.A beakers
Middle addition 0.2g soluble starches, and constantly stirred with glass bar, the first for making soluble starch be evenly dispersed in A beakers
In amide solution.Finally, the formamide solution in A beakers dissolved with soluble starch is added to temperature as 100 in multiple times on a small quantity
DEG C B beakers in formamide solution in dissolved.Dissolving is finished, and takes out B beakers natural cooling in atmosphere, and sealing is protected
Deposit.
PBS cushioning liquid is as obtained by being prepared following steps：
First with chloroazotic acid, (concentrated hydrochloric acid is 3 by volume with concentrated nitric acid:1 composition mixture) immersion two 100mL, one
The beaker that 1000mL volumetric flask, three volumes are 100mL 24 hours.Then, volumetric flask first is rinsed well with burning with running water
Cup, then with secondary water wash clean volumetric flask and beaker, store for future use.On electronic balance, disodium hydrogen phosphate is first weighed
14.196g, pour into the 100mL small beakers that numbering is A, adding secondary water and constantly being stirred with glass bar makes it completely molten
Solution, is then transferred into 100mL volumetric flasks, finally at the graduation mark with rubber head dropper constant volume to volumetric flask.It is labelled：Concentration
For 1mol/L disodium phosphate soln.Equally, 11.998g sodium dihydrogen phosphates are weighed on electronic balance, pour them onto numbering
For in B 100mL small beakers, then made with the secondary water dissolving phosphoric acid sodium dihydrogen of certain volume and constantly being stirred with glass bar
It is completely dissolved, and is then transferred into 100mL volumetric flasks, finally at the graduation mark with rubber head dropper constant volume to volumetric flask.
It is labelled：Concentration is 1mol/L sodium dihydrogen phosphate.Finally, 12mL disodium phosphate soln, 88mL phosphoric acid is taken
In the small beaker that dihydro sodium solution is C to numbering, constant volume is diluted into 1000mL volumetric flask.It is labelled：0.1mol/
L PBS cushioning liquid.
Embodiment 1
First on computers using the required figure of Corel DRAW Software on Drawing experiment.Wherein, circular sample application zone is straight
Footpath is 5.0mm, and the width and length in hough transform area are respectively that 3.0mm and 40.0mm, detection zone come with interval 3mm mark
Chi.After device pattern to be analyzed is drawn, a filter paper (9cm × 9cm, quantitative, middling speed) is positioned over the work of laser engraving machine
Make on platform, start instrument, by laser engraving machine, the paper microfluidic analysis required for experiment is cut out on quantitative, Medium speed filter paper
Device.In the detection zone of paper microfluidic analysis device, it is uniformly coated with what is configured by formamide solution with 1 μ L microsyringe
The μ L of 2% soluble starch solution 1, dry under field conditions (factors), be stored in the clean local back up without dust.
In 1.5mL centrifuge tube, the μ g/mL of certain volume 2 GOD solution is added with liquid-transfering gun first, is subsequently added into one
Determine the Ag of concentration+Solution.Then, having added the centrifuge tube of solution to be put into the water bath of 40 DEG C of constant temperature, it is allowed to react 15min.
After 15min, give again in centrifuge tube and add 20mM glucose solution, be then put into water bath and continue to react 30min.30min
Afterwards, to the IodineSodium Solution for continuously adding 0.5M in centrifuge tube, it is allowed to react 5min.After 5min, taken from constant water bath box
Go out centrifuge tube, be then added dropwise with liquid-transfering gun draw solution in the sample application zone of paper microfluidic analysis device.Then, analytical solution is in hair
The detection zone of paper microfluidic analysis device is flowed under capillary action, the 2% soluble starch solution that detection zone is fixed runs into
Elemental iodine in analytical solution, coloured track will be produced.Same experimental procedure, change Ag in reaction solution+Concentration
Carry out a series of experiment.Finally, by measuring the length of detection zone blueness track to Ag+Carry out quantitative detection.
Conclusion
It is 2 μ g/mL, Ag in optimal GOD concentration+Concentration is respectively blank (0) with 1mM, carrying out feasibility Experiment.It is real
It is as shown in Figure 2 to test result.It can be drawn by figure：Work as Ag+When concentration is 1mM, do not have in the detection zone of paper microfluidic analysis device
Generate blue trace；Work as Ag+When concentration is 0, uniform blue trace has been observed in the detection zone of paper microfluidic analysis device
Generation.Therefore, when GOD concentration is 2 μ g/mL, quantitative detection can be carried out to silver ion by paper microfluidic analysis device.
Based on paper microfluidic analysis device to Ag+Carry out quantitative detection
It is 2 μ g/mL, Ag in GOD concentration+When concentration is by 1nM-1mM (ten times of changes), a series of experiment has been carried out.
Among experiment, 2 μ g/mL GOD solution is first added in colorimetric cylinder, is subsequently added into certain density Ag+Solution；Then colorimetric
Pipe, which is placed in 40 DEG C of constant water bath box, reacts 15min.Afterwards, colorimetric cylinder is taken out, it is certain to being added in colorimetric cylinder with liquid-transfering gun
Volume 20mM glucose solution, react 30min；At this time not by Ag+The GOD of suppression is anti-by the hydrolysis for being catalyzed glucose solution
Should, reaction generation hydrogenperoxide steam generator.Then, then toward the IodineSodium Solution that 0.5M is added in colorimetric cylinder, the peroxidating in solution
In IodineSodium Solution redox reaction, reaction generation elemental iodine will occur for hydrogen.Finally, a small amount of reaction solution is pipetted with liquid-transfering gun
It is added dropwise in the sample application zone of paper microfluidic analysis device, is then printed in the blueness of the detection region measurement generation of paper microfluidic analysis device
Mark length.Experimental result is as shown in Figure 3.In figure 3 a, it can be seen that Ag+Concentration is respectively 10-4、10-6、10-8During mol/L,
The length for the blue trace that the detection zone of paper microfluidic analysis device detects is significantly different.Also, Ag+Concentration is smaller, in paper
The length for the blue trace that microfluidic analysis device detection zone detects is longer, their inversely proportional relations.In figure 3b, may be used
To find out：Detect the length and Ag of the blue trace of region measurement+Concentration between into good linear relationship.Work as Ag+Concentration exists
In the range of 1nM-1mM, corresponding equation of linear regression is Y=-3.3869+1.46131x (R2=0.93493).Work as Ag+Concentration is
During 1nM, the length that the detection zone of paper microfluidic analysis device detects the length of blue trace and blank sample detects is basic
Equally, do not distinguish significantly.Therefore, the detection lower limit of this method is about 1.0 × 10-9mol/L.Thus the method for seeing to find out
It can be used for detecting the Ag in solution well+。
Under above experiment condition, Ag is chosen+、Cd2+、Co2+、Cr2+、Cu2+、Fe2+、Hg2+、Mn2+、Ni2+、Pb2+、Zn2+Ten
One metal ion species carry out specificity experiments.Concentration wherein per metal ion species is all 100 μM.In experimentation, this is allowed
Ten metal ion species replace Ag+, respectively with GOD solution reactions；Ensuing experimental procedure and detection Ag+Operating procedure it is the same.
Testing result is as shown in Figure 4.It can be drawn by the data in figure, this ten metal ion species based on paper microfluidic analysis device to being determined
Amount detection Ag+Concentration do not influence substantially.Work as Ag+When concentration is 100 μM, in the detection region measurement of paper microfluidic analysis device
To the average length of blue trace be about 2.33mm (parallel laboratory test three times).It is but dense when ten species specificity metal ions
When degree is all 100 μM, 7- is all distributed in substantially in the length for the blue trace that the detection zone of paper microfluidic analysis device measures
Between 8mm.This just illustrates this method to Ag+Specificly-response.From this figure it can be seen that ten species specificity metal ions are dense
When degree is all 100 μM, all long in the blue contact length that paper microfluidic analysis device detection zone measures, it is equal to Ag+The blue contact length of detection zone when concentration is 100nM.Thus, can also draw, analysis method to Ag+Specificity choosing
Select.Therefore, it can be deduced that：This method strong interference immunity, it can be used for detecting the Ag in solution well+Concentration.
Embodiment 2
Analysis detection is carried out to the running water in laboratory with the paper microfluidic analysis device of design.In the analysis of running water
During, devise four groups of recovery testus.Wherein, the mark-on Ag in every group of recovery experiment+Concentration is different.Four kinds of mark-on Ag+
Solution is configured with running water, and its concentration is respectively：100nM、1μM、10μM、100μM.Experimental result is as shown in table 3.By table
Data can be drawn in 3：The paper microfluidic analysis device of exploitation can be used for detecting the Ag in running water well+.Wherein, add
Enter the Ag that normal concentration is 100nM+When, its rate of recovery is 93.57%, relative standard deviation 2.9%；Adding normal concentration is
1 μM of Ag+When, the rate of recovery 99.33%, relative standard deviation 5.7%；Add the Ag that normal concentration is 10 μM+When, recovery
Rate is 107.8%, relative standard deviation 5.0%；Add the Ag that normal concentration is 100 μM+When, the rate of recovery 103.25%,
Relative standard deviation is 5.7%.In summary, the analysis method can be applied preferably among the analysis of actual sample.
Table 3 carries out analysis detection to the silver ion in running water
The selective inhibitory based on Ag+ to GOD activity of this specific implementation；So that GOD is catalyzed glucose solution
Hydrolysis generation compare less hydrogen peroxide；The sodium iodide of hydrogenperoxide steam generator and reproducibility with oxidisability is molten
Redox reaction occurs for liquid, generates elemental iodine；The complex compound of iodo- 2% soluble starch solution reaction generation blueness.By with
Upper principle, the silver ion in solution is detected with telemetry using colorimetric method respectively.Experimental principle figure is as shown in Figure 1.
In figure, finite concentration, the glucose oxidase solution and Ag+ solution of certain volume are added first in test tubes, in a constant temperature
Its reaction a period of time is allowed under degree；Then, the glucose solution of certain volume is added, allows it to react the regular hour；Then, add
Enter the IodineSodium Solution of concentration known, allow it to react certain time；Finally, certain density soluble starch solution is added, is seen
Examine the change of solution colour；Meanwhile the solution of generation elemental iodine is added dropwise in the sample application zone of paper microfluidic analysis device.Finally,
By observing the change can of solution colour in test tubes, qualitatively the content of silver ion and measurement paper are micro- in analytical solution
The length of the blue trace of stream control analytical equipment detection zone generation carries out quantitative analysis to silver ion.Method do not need instrument set
Standby operation, cost is cheap, simple to operate.Result of study shows, this method qualitative detection to minimum concentration of silver ions be
100fM, the minimum concentration of silver ions quantitatively detected are 1nM.
Described above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (1)
- A kind of 1. inexpensive Ag+Detection method, it is characterised in that comprise the following steps：S1, first on computers using the figure needed for Corel DRAW Software on Drawing, wherein, circular sample application zone it is a diameter of 5.0mm, the width and length in hough transform area are respectively that 3.0mm and 40.0mm, detection zone come with interval 3mm scale；S2, after the completion for the treatment of graphic plotting, a 9cm × 9cm quantitative Medium speed filter paper is positioned over to the workbench of laser engraving machine On, start instrument, by laser engraving machine, required paper microfluidic analysis device is cut out on filter paper, in paper microfluidic analysis The detection zone of device, the μ of 2% soluble starch solution 1 configured by formamide solution is uniformly coated with 1 μ L microsyringe L, dry under field conditions (factors), be stored in the clean place without dust, it is standby；S3, in 1.5mL centrifuge tube, sequentially add the μ g/mL of certain volume 2 GOD solution and certain density Ag+Solution, so It is placed on afterwards in 40 DEG C of constant water bath box, after reacting 15min, 20mM glucose solution is added into centrifuge tube, is continued React 30min；Then 0.5M IodineSodium Solution is added into centrifuge tube, continues to react 5min；After the completion of S4, reaction, centrifuge tube is taken out from constant water bath box, is then added dropwise with liquid-transfering gun draw solution in step S2 institutes The sample application zone of the paper microfluidic analysis device obtained, analytical solution flow into the detection of paper microfluidic analysis device under capillarity Area, the 2% soluble starch solution that detection zone is fixed run into the elemental iodine in analytical solution, produce coloured track, lead to The length for crossing the coloured track of measurement detection zone is completed to Ag+Quantitative detection.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109839375A (en) * | 2019-01-30 | 2019-06-04 | 浙江工业大学 | A kind of paper substrate micro-fluidic chip and detection method for examination of glucose concentration |
| CN110596085A (en) * | 2019-09-03 | 2019-12-20 | 中山大学 | Distance measurement-based non-consumable paper chip and preparation method and application thereof |
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| CN104931489A (en) * | 2015-05-17 | 2015-09-23 | 桂林理工大学 | Quantitative analysis method for measuring rapid test strip on basis of iodine-starch variable-color distance |
| CN106872453A (en) * | 2017-01-11 | 2017-06-20 | 浙江大学 | Starch potassium iodide microchannel plastic sheeting detection strip and preparation method and purposes |
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| CN104931489A (en) * | 2015-05-17 | 2015-09-23 | 桂林理工大学 | Quantitative analysis method for measuring rapid test strip on basis of iodine-starch variable-color distance |
| CN106872453A (en) * | 2017-01-11 | 2017-06-20 | 浙江大学 | Starch potassium iodide microchannel plastic sheeting detection strip and preparation method and purposes |
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| CN109839375A (en) * | 2019-01-30 | 2019-06-04 | 浙江工业大学 | A kind of paper substrate micro-fluidic chip and detection method for examination of glucose concentration |
| CN110596085A (en) * | 2019-09-03 | 2019-12-20 | 中山大学 | Distance measurement-based non-consumable paper chip and preparation method and application thereof |
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