CN106093328B - A kind of Pb2+Pb in micro-fluidic detection chip and water sample2+Visible detection method - Google Patents
A kind of Pb2+Pb in micro-fluidic detection chip and water sample2+Visible detection method Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/18—Water
- G01N33/1813—Specific cations in water, e.g. heavy metals
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/02—Adapting objects or devices to another
- B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details
- B01L2200/027—Fluid interfacing between devices or objects, e.g. connectors, inlet details for microfluidic devices
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/12—Specific details about materials
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Abstract
Pb provided by the invention2+Micro-fluidic detection chip and including substrate, gel film, fixed column, showing rod and transparent cover plate, substrate is provided with the microchannel of the first microchannel~the 4th, intermediate axle passage, sample intake passage, sample output passage and indicator solution passage, some showing rods are provided with intermediate axle passage, showing rod forms showing rod's array in intermediate axle passage, gel film is arranged in fixed column, sample intake passage outlet respectively with first and the 4th microchannel inlet communication, first and the 4th the outlet of microchannel connected respectively with the outlet and injection port of intermediate axle passage, the outlet and injection port of intermediate axle passage respectively with second and the 3rd microchannel inlet communication, second and the 3rd the outlet of microchannel connected with the entrance of sample output passage, the outlet of indicator solution passage and the indicator solution inlet communication of intermediate axle passage, transparent cover plate is bonded with substrate and is integrated.Present invention also offers Pb in water sample2+Visible detection method, this method can be reduced to Pb in water sample2+Testing cost.
Description
Technical field
The invention belongs to the Pb based on intelligent gel2+Detection field, more particularly to a kind of Pb2+Micro-fluidic detection chip and
Pb in water sample2+Visible detection method.
Background technology
Lead ion is a kind of toxic heavy metal ion that significant damage can be caused to ecological environment and health, even
Micro lead ion also can produce huge harm to the nervous system of human body, digestive system, hemopoietic system, especially can be to children
Intelligence and grow and cause irreversible serious harm.Provided in GB25466-2010, the content of Pb in Drinking Water ion
4.83 × 10 must not be higher than-8Mol/L, the concentration of industrial wastewater ion must not be higher than 2.42 × 10-6Mol/L, thus may be used
Know, detect the lead ion of the super low concentration in the samples such as drinking water, industrial wastewater exactly, protected for health and environment
Shield all has very important significance.
At present, common lead ion detection method has atomic spectroscopy, electrochemical process and colorimetric method.Wherein, atom light
Spectrometry is to be realized using Atomic Absorption Spectrometer, AFS, icp mses etc. to ultralow dense
The detection of lead ion is spent, but this method needs to use the detection device of accurate costliness, and there is the deficiency that testing cost is high.Electricity
Chemical method is also required to using complicated instrument and equipment, and detecting step is cumbersome, needs professional to operate, thus is difficult to promote
Using.Peng Chi side etc. is based on colorimetric method, and concentration in water can be measured using 2 mercapto ethanol-sodium thiosulfate-nanogold composite membrane
For 5 × 10-8Mol/L lead ion (Peng Chifang, Xie Zhengjun, Wang Yayun, waits preparation and the lead ion meat of nanogold laminated films
Structure [J] analysis test journals of eye detection method, 2014,33 (10):1194-1198.), but colorimetric method judges with naked eyes,
Accurately quantitative detection can not be accomplished, and to realize quantitative detection, generally required by equipment such as the spectrophotometers of precision.
The content of the invention
It is an object of the invention to overcome the deficiencies of the prior art and provide a kind of Pb2+In micro-fluidic detection chip and water sample
Pb2+Visible detection method, with reduce to Pb in water sample2+Testing cost.
Pb provided by the invention2+Micro-fluidic detection chip, including substrate, gel film, fixed column, showing rod and transparency cover
Plate；
The substrate be provided with the first microchannel, the second microchannel, the 3rd microchannel, the 4th microchannel, intermediate axle passage,
Sample intake passage, sample output passage and indicator solution passage；First microchannel and the 4th microchannel axisymmetricly arrange, the second microchannel and
3rd microchannel axisymmetricly arranges that first microchannel, the second microchannel and the 3rd microchannel are equipped with slot segments, described
Some showing rods are provided with intermediate axle passage, showing rod forms at least one showing rod's array in intermediate axle passage, each instruction
Spacing in post array between adjacent showing rod is equal；
The fixed column is cylinder, and fixed column is arranged in the 4th microchannel, and the axis of fixed column is micro- logical perpendicular to the 4th
The bottom surface in road and intersecting with the center line of the 4th microchannel, the gel film is circular ring plate, and gel film is arranged in fixed column,
The material of gel film is poly- (hat of NIPA -co- benzo 18 6), when temperature is less than poly- (N- isopropyl acrylamides
During the hat Volume-phase transition temperature 6) of amine -co- benzo 18, the thickness and external diameter of gel film respectively with the depth of the 4th microchannel and
Width is identical, blocks the flow channels of the 4th microchannel, when temperature is higher than poly- (hat of NIPA -co- benzo 18 6)
During the Volume-phase transition temperature of microgel, gel film shrinks, and forms flow channels in the 4th microchannel, the horizontal stroke of the flow channels
Area of section is more than the cross-sectional area of the first microchannel slot segments, when containing Pb2+Water sample flow through the 4th microchannel, gel film choosing
It is complexed Pb to selecting property2+And generating body product swelling, reduce the flow channels of the 4th microchannel；
The sample intake passage exports the inlet communication with the first microchannel and the 4th microchannel respectively, and described first is micro- logical
The outlet in road and the 4th microchannel connects with the outlet and injection port of intermediate axle passage respectively, and the intermediate axle passage goes out sample
Mouthful and the injection port inlet communication with the second microchannel and the 3rd microchannel respectively, second microchannel and the 3rd microchannel
Outlet connects with the entrance of sample output passage, the outlet of the indicator solution passage and the indicator solution inlet communication of intermediate axle passage；
The transparent cover plate is bonded with substrate is formed as one Pb2+Micro-fluidic detection chip, transparent cover plate should be completely covered
Firmly all passages on substrate, transparent cover plate are relative with the entrance of the sample intake passage on substrate, sample output passage and indicator solution passage
Through hole should be located to be provided with, transparent cover plate in the through hole forms Pb after being bonded with substrate2+The injection port of micro-fluidic detection chip, go out
Sample mouth and indicator solution import.
Above-mentioned Pb2+In micro-fluidic detection chip, first microchannel, the second microchannel, the 3rd microchannel and the 4th are micro-
The width of passage is 150~500 μm, depth is 30~200 μm.
Above-mentioned Pb2+In micro-fluidic detection chip, slot segments on first microchannel, the second microchannel and the 3rd microchannel
Width be the first microchannel, 0.1~0.25 times of the second microchannel and the 3rd microchannel width, it is the first microchannel, second micro-
The depth of the slot segments depth phase with the first microchannel, the second microchannel and the 3rd microchannel respectively on passage and the 3rd microchannel
Deng.
Above-mentioned Pb2+In micro-fluidic detection chip, the first microchannel, the second microchannel, the 3rd microchannel on substrate, the 4th
The design principle of microchannel is：The shape of second microchannel and the 3rd microchannel should ensure that liquid flows through the second microchannel and the 3rd
During microchannel, the flow resistance in the second microchannel and the 3rd microchannel is equal, and to realize the second microchannel and the 3rd micro- logical
Flow resistance in road is equal and meets the annexation between each passage, it is desirable to which the second microchannel and the 3rd microchannel are in axle
It is arranged symmetrically；The shape of first microchannel and the 4th microchannel should meet claimed below：When gel film shrinks, gel film and
The cross-sectional area of the flow channels formed between four microchannel walls is more than the cross-sectional area of the slot segments of the first microchannel,
The flow resistance that flow resistance in the first microchannel is more than in the 4th microchannel should be met by being now passed through deionized water, be met
The condition simultaneously meets the annexation between each passage, preferably make the first microchannel and the 4th microchannel length,
Width and deep equality, axisymmetricly mode is arranged (except solidifying with the 4th microchannel at the slot segments of the first microchannel for the two
Shape at film is different outer, and the shape of the first microchannel and the 4th microchannel is axisymmetricly).As long as meet above-mentioned design
Principle, the first microchannel can be with equal with length, width, the depth of the second microchannel, can also be unequal, preferable mode
It is the first microchannel is also axisymmetricly arranged with the second microchannel.
Above-mentioned Pb2+In micro-fluidic detection chip, the width of the intermediate axle passage is 500~3000 μm, depth be 30~
200 μm, the showing rod is that diameter is 30~200 μm of cylinder, the spacing in showing rod's array between adjacent showing rod
For 30~100 μm.
Above-mentioned Pb2+In micro-fluidic detection chip, the fixed column is cylinder, a diameter of 4th microchannel width of fixed column
0.2~0.5 times.
Above-mentioned Pb2+In micro-fluidic detection chip, the substrate, fixed column and the material of showing rod are polydimethylsiloxanes
Alkane, the material of the cover plate can be the transparent materials such as glass or dimethyl silicone polymer.Existed using existing Soft lithograph technique
First microchannel, the second microchannel are set on substrate, the 3rd microchannel, the 4th microchannel, intermediate axle passage, sample intake passage, gone out
Sample passage, indicator solution passage, fixed column and showing rod, it specifically can refer to Qin, D.et al.Soft lithography for
Method in micro-and nanoscale patterning.Nature Protocols, 2010,5,491-502. is set
Put.
Above-mentioned Pb2+In micro-fluidic detection chip, the method that gel film is set in the 4th microchannel is：
1. it is monomer with NIPA, the hat of benzo 18 6, using azo diisobutyl amidine dihydrochloride as initiation
Agent, with N, N'- methylene-bisacrylamides are crosslinking agent, and the monomer, initiator, crosslinking agent and deionized water are well mixed
Form gel precursor liquid, in gel precursor liquid, the concentration of NIPA be 0.5~1.5mol/L, the hat of benzo 18 6 and
The mol ratio of NIPA is (1~5):20, the mol ratio of initiator and NIPA is 1:(5~
15), the mol ratio of crosslinking agent and NIPA is 1:(20~100)；
2. gel precursor liquid will be filled in the 4th microchannel, it is micro- logical that the shadow shield with circular loophole then is placed in into the 4th
The axis of loophole is set to be overlapped with the axis of fixed column above road, the diameter of the loophole and the width phase with the 4th microchannel
Deng, 1~15 DEG C with ultraviolet light through shadow shield loophole irradiate the 4th microchannel, trigger loophole at the 4th microchannel
Interior gel precursor liquid crosslinks reaction and forms gel film, and the gel forerunner in the 4th microchannel is then washed with deionized water
Liquid, that is, complete the setting of gel film in the 4th microchannel.
When gel film is set in the 4th microchannel, the shadow shield is hidden by being avoided that the material that ultraviolet passes through makes
Tabula rasa is preferably black film.
Present invention also offers Pb in a kind of water sample2+Visible detection method, this method using constant current conveying device, heat
Platform and above-mentioned Pb2+Micro-fluidic detection chip, the detection chip is placed in thermal station, by the injection port of detection chip and instruction
Liquid import is connected by pipe fitting with constant current conveying device respectively, and step is as follows：
1. using deionized water as blank sample, using constant current conveying device by blank sample with constant volume flow from entering
In sample mouth input detection chip, blank sample is discharged after chip from outlet after testing, using constant current conveying device by indicator solution
Inputted with constant volume flow from indicator solution import in detection chip, indicator solution in the intermediate axle passage of chip to be detected
After interface location is stable, using the number for the showing rod that liquid covering is instructed in amplification sem observation intermediate axle passage, the instruction
Liquid is formulated by water-soluble dye and deionized water；
2. by step 1. in blank sample replace with a series of Pb2+Standard specimen known to concentration, the operation of repeat step 1.,
Obtain being instructed to the number of the showing rod of liquid covering corresponding to a series of standard specimens, calculate when inputting each standard specimen relative to input blank
The changing value of showing rod's number of liquid covering is instructed to during sample, with Pb in standard specimen2+Concentration is ordinate, covered with being instructed to liquid
The changing value of showing rod's number be abscissa drawing curve, determine Pb2+Concentration and the showing rod's number for being instructed to liquid covering
The conversion relation formula of purpose changing value；
3. step 1. in blank sample replace with sample to be tested, the operation of repeat step 1., obtain inputting sample to be tested
When be instructed to liquid covering showing rod number, be instructed to liquid and cover when when calculating input sample to be tested relative to input blank sample
The changing value of showing rod's number of lid, the Pb 2. determined according to step2+Concentration and the change for being instructed to showing rod's number that liquid covers
The conversion relation formula of change value calculates Pb in testing sample2+Concentration；
Step 2., 3. in, after completing the detection to each standard specimen or sample to be tested, deionization is inputted into detection chip
Water cleans gel film, in cleaning process, the temperature of thermal station is risen into 55~60 DEG C and is incubated 3~5min, is then down to 20~25
DEG C and be incubated 3~5min, repeat it is foregoing heating cleaning and cooling cleaning operation after removing the metal ion in gel film
Next standard specimen or sample is inputted again to be detected；
Step 1.~3. in, during input blank sample, standard specimen and sample to be tested are detected, control thermal station
Temperature for 33~35 DEG C and keeping temperature it is constant.
Pb in above-mentioned water sample2+Visible detection method in, into detection chip input blank sample, standard specimen or to be tested
During sample, control input flow is 200~8000 μ L/h, into detection chip input indicator solution when, control input flow be 50~
2000μL/h。
Pb in water sample of the present invention2+Visible detection method principle it is as follows：
In detection chip of the present invention, gel film is fixed in the fixed column of the 4th microchannel, when temperature is higher than poly-
During the Volume-phase transition temperature of (hat of NIPA -co- benzo 18 6) microgel, gel film shrinks, now gel film
The area of the flow channels formed between the 4th microchannel wall is more than the area of passage of the slot segments of the first microchannel, to institute
State and deionized water is passed through in detection chip, by now the first microchannel, the second microchannel, the 3rd microchannel, the 4th microchannel
Flow resistance is denoted as R respectively1、R2、R3、RX, because the second microchannel and the 3rd microchannel axisymmetricly arrange that is, the two are micro-
Equal (the R of flow resistance of passage2=R3), thus now the relation between the flow resistance of four microchannels is R1R3>>R2RX,
Now the liquid in intermediate axle passage can flow to outlet by the injection port of intermediate axle passage, as shown in Fig. 4 (a)；When containing low dense
Spend Pb2+Water sample when flowing through gel film, the group of 18- crown-s 6 of poly- (hat of NIPA -co- benzo 18 6) can select
It is complexed lead ion and forms powered complex compound to property, the electrostatic repulsion between charged complex group can causes macromolecule
Chain-unfolding, and then cause gel film that volume sweell(ing) occurs, meanwhile, poly- (hat of NIPA -co- benzo 18 6)
After the group of 18- crown-s 6 complexing lead ion the osmotic pressure of gel film can be caused to increase, cause gel film that water absorption and swelling, gel film occurs
After swelling, the flow channels of the 4th microchannel reduce, as shown in Fig. 4 (b), the now flow resistance R ' of the 4th microchannelX>RX, make
The flow velocity for obtaining the liquid flowed in intermediate axle passage from injection port to outlet reduces；As Pb in water sample2+Concentration further increase
When big, the flow resistance R " of the 4th microchannelXContinue to increase, cause R1R3<R2R”X, now liquid flow direction is sent out in intermediate axle passage
Raw transformation, flows to injection port, as shown in Fig. 4 (c) by outlet.Due to the degree and Pb of gel film volume sweell(ing)2+Concentration phase
Close, and the number that the showing rod of liquid covering is instructed in detection chip intermediate axle passage is related to the degree of swelling of gel film, and
The change of gel film degree of swelling can make the first microchannel and the relative size of the flow resistance in the 4th microchannel change,
Therefore the change of showing rod's number of liquid covering is instructed in intermediate axle passage of the method for the invention by determining detection chip
Change value is realized to Pb in water sample2+Quantitative determination.
Compared with prior art, the invention has the advantages that：
1. Pb provided by the invention2+Micro-fluidic detection chip is a kind of new lead ion detection chip, the detection chip
Detection to lead ion is realized based on intelligent gel and Wheatstone bridge principle, gel film therein can specifically identify Pb2+
And generating body product swelling, and cause the flow channels of the 4th microchannel to reduce, cause the flow resistance of the 4th microchannel to increase, enter
And the change for causing the flow velocity of liquid in intermediate axle passage even to flow to, therefore, by the detection chip of the present invention with using perseverance
Flow conveying device with thermal station to coordinate, showing rod's number of liquid covering is instructed to by observing in the detection chip intermediate axle passage
Changing value, you can realize that the visualization to lead ion in water sample quantitatively detects.
2. present invention also offers one kind to detect Pb in water sample2+New method, the detection limit as little as 10 of this method-10mol/
L, 10 can be realized-10~10-4The detection of the other lead ion of mol/L concentration levels, detection limit is low, detection range is wide.
3. the detection chip of the present invention is used cooperatively and realized by the method for the invention with constant current conveying device and thermal station
Detection to lead ion in water sample, these devices are common device, cheap, compared with prior art, without using each
The aided-detection device of class costly, without professional and technical personnel's operating instrument, the method for the invention, which has, to be parsed into
The advantage of this application cheap, simple to operate, applied widely and easy to spread.
Brief description of the drawings
Fig. 1 is Pb of the present invention2+The structural representation of micro-fluidic detection chip；
Fig. 2 is the Pb in Fig. 12+Partial enlarged drawing in 4th microfluidic channel of micro-fluidic detection chip at gel film；
Fig. 3 is the Pb in Fig. 12+Office at the slot segments of first, second, third microfluidic channel of micro-fluidic detection chip
Portion's enlarged drawing；
Fig. 4 is using detection chip of the present invention detection Pb2+Principle schematic；
Fig. 5 is the schematic diagram that gel film is set in the 4th microchannel；
Fig. 6 is the microscope photograph of the forming process of gel film in the 4th microchannel, wherein figure A~D be followed successively by be passed through it is solidifying
Before glue precursor liquid, be passed through gel precursor liquid after, ultraviolet light forms gel film and washes away the microscope after gel precursor liquid
Picture；
Fig. 7 is that embodiment 4 is passed through different Pb2+After the sample solution of concentration, the interface location of indicator solution in intermediate axle passage
Microscope photograph, wherein figure A~F is followed successively by and is passed through deionized water, 10-10、10-8、10-7、10-6、10-5Mol/L Pb2+Sample
Microscope photograph during product solution；
Fig. 8 is the working curve that embodiment 4 is drawn；
Fig. 9 is the method for the invention detection Li+、Na+、K+、Mg2+、Ca2+、Sr2+、Ba2+、Cr3+、Co2+、Ni2+、Cu2+、
Zn2+、Cd2+、Pb2+When intermediate axle passage in indicator solution interface location microscope photograph；
Figure 10 is the method for the invention detection Li+、Na+、K+、Mg2+、Ca2+、Sr2+、Ba2+、Cr3+、Co2+、Ni2+、Cu2+、
Zn2+、Cd2+、Pb2+When intermediate axle passage in be instructed to liquid covering showing rod's number changing value block diagram；
In figure, the microchannel of 1-substrate, 1-1-first, the microchannels of 1-2-second, the microchannel 1-4-the 4th of 1-3-the 3rd
Microchannel, 1-5-intermediate axle passage, 1-5-1-outlet, 1-5-2-injection port, 1-6-sample intake passage, 1-7-go out sample and led to
Road, 1-8-indicator solution passage, 1-9-slot segments, 2-gel film, 3-fixed column, 4-showing rod, 5-transparent cover plate, 6-
Shadow shield.
Embodiment
By the following examples to Pb of the present invention2+Pb in micro-fluidic detection chip and water sample2+Visual retrieval
Method is described further.
Embodiment 1
In the present embodiment, Pb2+The structure of micro-fluidic detection chip is as shown in Figures 1 to 3, including substrate 1, gel film 2, fixed
Post 3, showing rod 4 and transparent cover plate 5.
The substrate 1 is provided with the first microchannel 1-1, the second microchannel 1-2, the 3rd microchannel 1-3, the 4th microchannel 1-
4th, intermediate axle passage 1-5, sample intake passage 1-6, sample output passage 1-7 and indicator solution passage 1-8；First microchannel 1-1 and the 4th is micro-
Passage 1-4 axisymmetricly arranges that the second microchannel 1-2 and the 3rd microchannel 1-3 are axisymmetricly arranged, the first microchannel 1-1 with
Second microchannel 1-2 also axisymmetricly arranges that the first microchannel 1-1, the second microchannel 1-2 and the 3rd microchannel 1-3 are equal
Provided with slot segments 1-9；First microchannel, the second microchannel, the 3rd microchannel, the width of the 4th microchannel are 300 μm, depth
It it is 70 μm, the slot segments 1-9 set on the first microchannel, the second microchannel and the 3rd microchannel width is 50 μm, depth
Degree is 70 μm, length is 300 μm；The width of the intermediate axle passage 1-5 is 1100 μm, depth is 70 μm, and intermediate axle is led to
Be provided with road 1-5 it is some be shaped as cylinder, a diameter of 100 μm of showing rod 4, showing rod 4 is in intermediate axle passage 1-5 close to going out
The position of sample mouth and injection port forms Liang Ge showing rods array, and three rows (often arranging five) instruction is included in each showing rod's array
Post, the spacing between adjacent showing rod in Liang Ge showing rods array is 100 μm.
The fixed column 3 is a diameter of 100 μm of cylinder, and fixed column is arranged in the 4th microchannel 1-4 and fixed column
Axis intersects perpendicular to the bottom surface of the 4th microchannel and with the center line of the 4th microchannel, and the gel film 2 is circular ring plate, gel
Piece is arranged in fixed column 3, and the material of gel film 2 is poly- (hat of NIPA -co- benzo 18 6), when temperature is less than
During the Volume-phase transition temperature of poly- (NIPA -co- benzo 18 hat 6), the thickness and external diameter of gel film 2 respectively with
The depth and width of 4th microchannel (1-4) are identical, block the flow channels of the 4th microchannel, when temperature is higher than poly- (N- isopropyls
When base acrylamide -co- benzo 18 is preced with 6) the Volume-phase transition temperature of microgel, gel film 2 shrinks, in the 4th microchannel 1-4
Middle formation flow channels, the cross-sectional areas of the flow channels are more than the first microchannel slot segments 1-9 cross-sectional area, when containing
Pb2+Water sample flow through the 4th microchannel 1-4, gel film is optionally complexed Pb2+And generating body product swelling, make the 4th microchannel
1-4 flow channels reduce.
The sample intake passage 1-6's exports the inlet communication with the first microchannel 1-1 and the 4th microchannel 1-4 respectively, institute
That states the first microchannel 1-1 and the 4th microchannel 1-4 exports the outlet 1-5-1 and injection port with intermediate axle passage 1-5 respectively
1-5-2 is connected, the outlet 1-5-1 and injection port 1-5-2 of the intermediate axle passage 1-5 respectively with the second microchannel 1-2 and the
Three microchannel 1-3 inlet communication, the second microchannel 1-2 and the 3rd microchannel 1-3 outlet and entering for sample output passage 1-7
Mouth connection, the outlet of the indicator solution passage 1-8 and intermediate axle passage 1-5 indicator solution inlet communication.
The transparent cover plate 5, substrate 1, fixed column 3 and the material of showing rod 4 are dimethyl silicone polymer (PDMS), thoroughly
Bonding is formed as one Pb after bright cover plate 5 and substrate 1 are handled by plasma bonder2+Micro-fluidic detection chip, transparent cover plate
All passages on substrate, transparent cover plate and sample intake passage 1-6, sample output passage 1-7 and the instruction on substrate should be covered all
Liquid passage 1-8 entrance corresponding section is provided with through hole, and transparent cover plate in the through hole forms Pb after being bonded with substrate2+It is micro-fluidic
Injection port, outlet and the indicator solution import of detection chip.
With reference to Qin, D.et al.Soft lithography for micro-and nanoscale
Method in patterning.Nature Protocols, 2010,5,491-502. set on substrate the first microchannel 1-1,
Second microchannel 1-2, the microchannel 1-4 of the 3rd microchannel 1-3 the 4th, intermediate axle passage 1-5, sample intake passage 1-6, sample output passage 1-
7th, indicator solution passage 1-8, fixed column 3 and showing rod 4.
The schematic diagram of gel film is set to see Fig. 5 in the 4th microchannel 1-4, step is as follows；
1. it is monomer with NIPA, phendioxin 8- hats 6, using azo diisobutyl amidine dihydrochloride as initiation
Agent, with N, N'- methylene-bisacrylamides are crosslinking agent, and the monomer, initiator, crosslinking agent and deionized water are added into container
In, well mixed to form gel precursor liquid, in gel precursor liquid, the concentration of NIPA is 1.0mol/L, benzo
The mol ratio of 18 hats 6 and NIPA is 3:20, the mol ratio of initiator and NIPA is 1:5,
The mol ratio of crosslinking agent and NIPA is 1:50；
2. the circular light penetrating slit of a diameter of 300 μm of setting one forms shadow shield on black film, from without gel film
Gel precursor liquid is passed through in the injection port of the micro-fluidic chip chip, makes to be full of gel precursor liquid in the 4th microchannel 1-4, so
The shadow shield 6 is placed in above the 4th microchannel 1-4 afterwards, the axis and the loophole of the shadow shield 6 for making fixed column 3 are hung down
The center of circle straight and through loophole, the diameter of loophole is equal with the 4th microchannel 1-4 width, 5 DEG C with ultraviolet light from saturating
The 4th microchannel 1-4 is irradiated in the top of tabula rasa through the loophole of shadow shield, triggers in the 4th microchannel 1-4 at loophole
Gel precursor liquid crosslinks reaction and forms gel film 2, and the gel forerunner in the 4th microchannel 1-4 is then washed with deionized water
Liquid, that is, complete the setting of gel film 2 in the 4th microchannel 1-4.Fig. 6 is the microscope photograph of the forming process of gel film.
2. the circular light penetrating slit of a diameter of 300 μm of setting one forms shadow shield on black film, from without gel film
Gel precursor liquid is passed through in the injection port of the micro-fluidic chip chip, makes to be full of gel precursor liquid in the 4th microchannel 1-4, then
The shadow shield 6 of loophole, which is placed in above the 4th microchannel 1-4, makes the axis of loophole be overlapped with the axis of fixed column 3, at 5 DEG C
The 4th microchannel 1-4 is irradiated with the loophole of ultraviolet light through shadow shield, is triggered solidifying in the 4th microchannel 1-4 at loophole
Glue precursor liquid crosslinks reaction and forms gel film 2, and the gel forerunner in the 4th microchannel 1-4 is then washed with deionized water
Liquid, that is, complete the setting of gel film 2 in the 4th microchannel 1-4.Fig. 6 is the microscope photograph of the forming process of gel film.
Embodiment 2
In the present embodiment, Pb2+Each microchannel and fixed column and showing rod in the structure of micro-fluidic detection chip, substrate
Preparation method, the method for setting gel film is essentially identical in the 4th microchannel, and difference is：It is first microchannel, second micro-
Passage, the 3rd microchannel, the width of the 4th microchannel are 500 μm, depth is 200 μm, the first microchannel, the second microchannel
Width with the slot segments 1-9 set on the 3rd microchannel is 50 μm, depth is 200 μm, length is 300 μm, and the 4th
A diameter of 250 μm of fixed column in microchannel, intermediate axle passage 1-5 width is 3000 μm, depth is 200 μm, showing rod 4
A diameter of 200 μm, showing rod 4 forms Liang Ge showing rods battle array in intermediate axle passage 1-5 close to the position of outlet and injection port
Row, include three rows (often arranging ten) showing rod in each showing rod's array, in Liang Ge showing rods array between adjacent showing rod
Spacing be 100 μm.In the gel precursor aqueous solution, the concentration of NIPA is 0.5mol/L, and benzo 18 is preced with 6
Mol ratio with NIPA is 1:20, initiator azo diisobutyl amidine dihydrochloride and N- isopropyl acrylamides
The mol ratio of amine is 1:10, the mol ratio of crosslinking agent N, N'- methylene-bisacrylamide and NIPA is 1:
100, when gel is set in the 4th microchannel, irradiate the 4th microchannel in 15 DEG C of loopholes with ultraviolet light through shadow shield.
Embodiment 3
In the present embodiment, Pb2+Each microchannel and fixed column and showing rod in the structure of micro-fluidic detection chip, substrate
Preparation method, the method for setting gel film is essentially identical in the 4th microchannel, and difference is：It is first microchannel, second micro-
Passage, the 3rd microchannel, the width of the 4th microchannel are 150 μm, depth is 30 μm, the first microchannel, the second microchannel
And the width of the 3rd slot segments on microchannel is 38 μm, depth is 30 μm, length is 300 μm, in the 4th microchannel
A diameter of 30 μm of fixed column, intermediate axle passage 1-5 width is 500 μm, depth is 30 μm, a diameter of 30 μm of showing rod 4,
Showing rod 4 forms showing rod's array in intermediate axle passage 1-5, and showing rod's array includes 12 rows (often arranging seven)
Showing rod, the spacing in showing rod's array between adjacent showing rod is 30 μm.In the gel precursor aqueous solution, N- isopropyls third
The concentration of acrylamide is 1.5mol/L, and the mol ratio of the hat of benzo 18 6 and NIPA is 5:20, initiator azo two
The mol ratio of isobutyl amidine dihydrochloride and NIPA is 1:15, crosslinking agent N, N'- methylene-bisacrylamide
Mol ratio with NIPA is 1:20, when gel is set in the 4th microchannel, is passed through and hidden with ultraviolet light at 1 DEG C
The loophole of tabula rasa irradiates the 4th microchannel.
Embodiment 4
In the present embodiment, Pb is prepared2+Concentration is 1 × 10-8Mol/L is examined as sample to be tested using the method for the invention
The Pb surveyed in the sample to be tested2+Concentration, this method use the Pb in syringe pump, thermal station and embodiment 12+Micro-fluidic detection core
Piece, the detection chip is placed in thermal station, the injection port of detection chip and indicator solution import are passed through into pipe fitting and injection respectively
Pump connects, and step is as follows：
1. using deionized water as blank sample, using syringe pump by blank sample with 2000 μ L/min constant volume flow
Inputted from injection port in detection chip, blank sample is discharged after chip from outlet after testing, using syringe pump by indicator solution with
500 μ L/min constant volume flow is inputted in detection chip from indicator solution import, and after 10min, the intermediate axle of detection chip is led to
The interface location of indicator solution reaches stable state in road, using the instruction that liquid covering is instructed in amplification sem observation intermediate axle passage
The number of post, is denoted as N0, the indicator solution is contaminated by methylene blue and deionized water is formulated；In the step, thermal station is controlled
Temperature is maintained at 32 DEG C.
2. prepare Pb with deionized water2+Concentration is respectively 1 × 10-10mol/L、1×10-8mol/L、1×10-7mol/L、1
×10-6mol/L、1×10-5Mol/L standard specimen, 1#~5# standard specimens are denoted as respectively.
Blank sample in using 1#~5# standard specimens replacement step successively 1., the operation of repeat step 1., the centre that will be measured
The number that the showing rod of liquid covering is instructed in bridge passage is denoted as N successively1, N2, N3, N4, N5, it is passed through blank sample and 1#~5# marks
During sample, after the interface location of indicator solution in intermediate axle passage is stable, the interface location figure of indicator solution is as shown in fig. 7, calculate defeated
Relative to the changing value Δ for showing rod's number that liquid covering is instructed in intermediate axle passage during input blank sample when entering each standard specimen
Ni=| N0-Ni|, i=1,2,3,4,5, with Pb in standard specimen2+Concentration is ordinate, to be instructed to the showing rod number of liquid covering
Changing value is abscissa drawing curve, as shown in figure 8, obtaining Pb2+Showing rod number of the concentration with being instructed to liquid covering
The conversion relation formula of changing value is C=8 × 10-18×(ΔN)9.08, in formula, C Pb2+Concentration, unit mol/L, Δ N are quilt
The changing value of showing rod's number of indicator solution covering, unit are individual；The step controls when being detected to 1#~5# standard specimens
The temperature of thermal station is maintained at 32 DEG C.
In the step, after completing the detection to each standard specimen, deionized water cleaning gel film is passed through into detection chip,
In cleaning process, the temperature of thermal station is risen to 55 DEG C and is incubated 3min, be then down to 25 DEG C and be incubated 3min, repeat foregoing heating
With the operation of cooling, until being passed through deionized water into detection chip according to the operation of step 1., it is instructed in intermediate axle passage
The showing rod's quantity and N of liquid covering0It is identical, illustrate that the lead ion in now gel film removes completely, then input next standard specimen
Or sample is detected.
3. using sample to be tested replace step 1. in blank sample, the operation of repeat step 1., measure be passed through it is to be tested
The number of the showing rod of liquid covering is instructed to during sample in intermediate axle passage, relative to input blank examination when calculating input sample to be tested
The changing value of showing rod's number of liquid covering, the Pb 2. determined according to step are instructed to during sample2+Concentration is with being instructed to liquid covering
The conversion relation formula of the changing value of showing rod's number calculates Pb in testing sample2+Concentration, it is as a result 1 × 10-8mol/L。
Embodiment 5
In the present embodiment, liquid covering is instructed in intermediate axle passage when investigating the method for the invention detection different kinds of ions
The situation of showing rod's number of variations value.Use the Pb in syringe pump, thermal station and embodiment 12+Micro-fluidic detection chip, by described in
Detection chip is placed in thermal station, and the injection port of detection chip and indicator solution import are connected by pipe fitting with syringe pump respectively, tool
Body step is as follows：
1. using deionized water as blank sample, using syringe pump by blank sample with 2000 μ L/min constant volume flow
Inputted from injection port in detection chip, blank sample is discharged after chip from outlet after testing, using syringe pump by indicator solution with
500 μ L/min constant volume flow is inputted in detection chip from indicator solution import, and after 10min, the intermediate axle of detection chip is led to
The interface location of indicator solution reaches stable state in road, using the instruction that liquid covering is instructed in amplification sem observation intermediate axle passage
The number of post, is denoted as N0, the indicator solution is contaminated by methylene blue and deionized water is formulated；In the step, thermal station is controlled
Temperature is maintained at 32 DEG C.
2. compound concentration is 1 × 10 respectively-6Mol/L Li+、Na+、K+、Mg2+、Ca2+、Sr2+、Ba2+、Cr3+、Co2+、Ni2 +、Cu2+、Zn2+、Cd2+、Pb2+Solution, it is denoted as 1#~14# samples；
Blank sample in using 1#~14# samples replacement step successively 1., the operation of repeat step 1., record input each examination
The number of the showing rod of liquid covering is instructed in intermediate axle passage during sample, when the interface location of indicator solution in intermediate axle passage is steady
After fixed, the interface location figure of indicator solution is as shown in figure 9, the step when being detected to 1#~14# samples, controls thermal station
Temperature is maintained at 32 DEG C.
In the step, after completing the detection to each standard specimen, deionized water cleaning gel film is passed through into detection chip,
In cleaning process, the temperature of thermal station is risen to 50 DEG C and is incubated 5min, be then down to 20 DEG C and be incubated 5min, repeat foregoing heating
With the operation of cooling, until being passed through deionized water into detection chip according to the operation of step 1., it is instructed in intermediate axle passage
The showing rod's quantity and N of liquid covering0It is identical, illustrate that the metal ion in now gel film removes completely, then input next examination
Sample is detected.
Relative to the finger that liquid covering is instructed in intermediate axle passage during input blank sample when calculating input 1#~14# samples
Show post number of variations value, as a result as shown in Figure 10, as shown in Figure 10, except Ba2+、Pb2+Outside, quilt of other ions under the concentration
Showing rod's number of variations value of indicator solution covering is 0, without interference with Pb2+Measure, and determine Ba2+When be instructed to liquid covering
Showing rod's number of variations value and Pb2+When be instructed to the showing rod number of variations value of liquid covering and differ more than 10 times, will not also do
Disturb Pb2+Measure.
Claims (9)
- A kind of 1. Pb2+Micro-fluidic detection chip, it is characterised in that including substrate (1), gel film (2), fixed column (3), showing rod And transparent cover plate (5) (4)；The substrate (1) is provided with the first microchannel (1-1), the second microchannel (1-2), the 3rd microchannel (1-3), the 4th micro- logical Road (1-4), intermediate axle passage (1-5), sample intake passage (1-6), sample output passage (1-7) and indicator solution passage (1-8)；First is micro- logical Road (1-1) and the 4th microchannel (1-4) axisymmetricly arrange that the second microchannel (1-2) and the 3rd microchannel (1-3) are axisymmetricly Arrangement, first microchannel (1-1), the second microchannel (1-2) and the 3rd microchannel (1-3) are equipped with slot segments (1-9), institute State and some showing rods (4) are provided with intermediate axle passage (1-5), showing rod (4) forms at least one in intermediate axle passage (1-5) Showing rod's array, the spacing in each showing rod's array between adjacent showing rod are equal；The fixed column (3) is cylinder, and fixed column is arranged in the 4th microchannel (1-4), and the axis of fixed column is perpendicular to the 4th The bottom surface of microchannel and intersect with the center line of the 4th microchannel, the gel film (2) is circular ring plate, and gel film is arranged on fixation On post (3), the material of gel film (2) is poly- (hat of NIPA -co- benzo 18 6), and when temperature, (N- is different less than poly- During the hat Volume-phase transition temperature 6) of propylacrylamide -co- benzo 18, the thickness and external diameter of gel film (2) are respectively with the 4th The depth and width of microchannel (1-4) are identical, block the flow channels of the 4th microchannel, when temperature is higher than poly- (N- isopropyls third When acrylamide -co- benzo 18 is preced with 6) the Volume-phase transition temperature of microgel, gel film (2) shrinks, in the 4th microchannel (1-4) Middle formation flow channels, the cross-sectional area of the flow channels are more than the cross-sectional area of the first microchannel slot segments (1-9), when Containing Pb2+Water sample flow through the 4th microchannel (1-4), gel film is optionally complexed Pb2+And generating body product swelling, make the 4th micro- The flow channels of passage (1-4) reduce；The outlet of the sample intake passage (1-6) respectively with the first microchannel (1-1) and the inlet communication of the 4th microchannel (1-4), The outlet of first microchannel (1-1) and the 4th microchannel (1-4) outlet (1-5- with intermediate axle passage (1-5) respectively 1) connected with injection port (1-5-2), the outlet (1-5-1) and injection port (1-5-2) of the intermediate axle passage (1-5) respectively with Second microchannel (1-2) and the inlet communication of the 3rd microchannel (1-3), second microchannel (1-2) and the 3rd microchannel (1- 3) outlet connects with the entrance of sample output passage (1-7), outlet and the intermediate axle passage (1-5) of the indicator solution passage (1-8) Indicator solution inlet communication；The transparent cover plate (5) is bonded with substrate (1) is formed as one Pb2+Micro-fluidic detection chip, transparent cover plate should cover completely All passages on substrate are covered, transparent cover plate leads to the sample intake passage (1-6) on substrate, sample output passage (1-7) and indicator solution The entrance corresponding section in road (1-8) is provided with through hole, and transparent cover plate in the through hole forms Pb after being bonded with substrate2+Micro-fluidic inspection Survey injection port, outlet and the indicator solution import of chip；The method of setting gel film is in the 4th microchannel (1-4)：1. it is monomer with NIPA, the hat of benzo 18 6, using azo diisobutyl amidine dihydrochloride as initiator, with N, N'- methylene-bisacrylamide are crosslinking agent, and the monomer, initiator, crosslinking agent and deionized water are well mixed and formed Gel precursor liquid, in gel precursor liquid, the concentration of NIPA is 0.5~1.5mol/L, and the hat 6 of benzo 18 is different with N- The mol ratio of propylacrylamide is (1~5):20, the mol ratio of initiator and NIPA is 1:(5~15), The mol ratio of crosslinking agent and NIPA is 1:(20~100)；2. gel precursor liquid will be filled in the 4th microchannel (1-4), the shadow shield (6) with circular loophole is then placed in the 4th The axis of loophole is overlapped with the axis of fixed column (3) above microchannel (1-4), the diameter of the loophole with it is the 4th micro- The width of passage (1-4) is equal, irradiates the 4th microchannel (1-4) in 1~15 DEG C of loophole with ultraviolet light through shadow shield, draws The gel precursor liquid in the 4th microchannel (1-4) at hair loophole crosslinks reaction and forms gel film (2), then spend from The gel precursor liquid that son washing is gone in the 4th microchannel (1-4), that is, complete the setting of gel film (2) in the 4th microchannel (1-4).
- 2. Pb according to claim 12+Micro-fluidic detection chip, it is characterised in that first microchannel (1-1), second micro- The width of passage (1-2), the 3rd microchannel (1-3) and the 4th microchannel (1-4) is 150~500 μm, depth is 30~200 μm.
- 3. Pb according to claim 1 or claim 22+Micro-fluidic detection chip, it is characterised in that first microchannel (1-1), The width of slot segments (1-9) is the first microchannel (1-1), the second microchannel on two microchannels (1-2) and the 3rd microchannel (1-3) 0.1~0.25 times of (1-2) and the 3rd microchannel (1-3) width, the first microchannel (1-1), the second microchannel (1-2) and the 3rd The depth of slot segments (1-9) is micro- logical with the first microchannel (1-1), the second microchannel (1-2) and the 3rd respectively on microchannel (1-3) The deep equality in road (1-3).
- 4. Pb according to claim 1 or claim 22+Micro-fluidic detection chip, it is characterised in that the width of the intermediate axle passage (1-5) It is 30~200 μm to spend for 500~3000 μm, depth, and the showing rod (4) is that diameter is 30~200 μm of cylinder, the instruction Spacing in post array between adjacent showing rod is 30~100 μm.
- 5. Pb according to claim 1 or claim 22+Micro-fluidic detection chip, it is characterised in that the fixed column (3) is cylinder, Gu 0.2~0.5 times of a diameter of 4th microchannel (1-4) width of fixed column.
- 6. Pb according to claim 1 or claim 22+Micro-fluidic detection chip, it is characterised in that the substrate (1), fixed column (3) and The material of showing rod (4) is dimethyl silicone polymer.
- 7. Pb according to claim 12+Micro-fluidic detection chip, it is characterised in that the shadow shield (6) is ultraviolet by being avoided that The material that line passes through makes.
- A kind of 8. Pb in water sample2+Visible detection method, it is characterised in that this method using constant current conveying device, thermal station and Pb described in any claim in claim 1 to 72+Micro-fluidic detection chip, the detection chip is placed in thermal station, will examined The injection port and indicator solution import for surveying chip are connected by pipe fitting with constant current conveying device respectively, and step is as follows：1. using deionized water as blank sample, using constant current conveying device by blank sample with constant volume flow from injection port Input in detection chip, blank sample is discharged after chip from outlet after testing, using constant current conveying device by indicator solution with perseverance Fixed volume flow is inputted in detection chip from indicator solution import, the interface position of indicator solution in the intermediate axle passage of chip to be detected After putting stabilization, using the number for the showing rod that liquid covering is instructed in amplification sem observation intermediate axle passage, the indicator solution is by water Soluble dye and deionized water are formulated；2. by step 1. in blank sample replace with a series of Pb2+Standard specimen known to concentration, the operation of repeat step 1., is obtained The number of the showing rod of liquid covering is instructed to corresponding to a series of standard specimens, is calculated when inputting each standard specimen relative to input blank sample When be instructed to liquid covering showing rod's number changing value, with Pb in standard specimen2+Concentration be ordinate, be instructed to liquid covering finger The changing value for showing post number is abscissa drawing curve, determines Pb2+Showing rod number of the concentration with being instructed to liquid covering The conversion relation formula of changing value；3. step 1. in blank sample replace with sample to be tested, the operation of repeat step 1., quilt when obtaining inputting sample to be tested The number of the showing rod of indicator solution covering, liquid covering is instructed to when calculating when inputting sample to be tested relative to input blank sample The changing value of showing rod's number, the Pb 2. determined according to step2+Concentration and the changing value for being instructed to showing rod's number that liquid covers Conversion relation formula calculate testing sample in Pb2+Concentration；Step 2., 3. in, after completing the detection to each standard specimen or sample to be tested, into detection chip input deionized water it is clear Wash gel film, in cleaning process, the temperature of thermal station is risen to 55~60 DEG C and is incubated 3~5min, is then down to 20~25 DEG C simultaneously 3~5min is incubated, the operation for repeating foregoing heating cleaning and cooling cleaning is defeated again after removing the metal ion in gel film Enter next standard specimen or sample to be detected；Step 1.~3. in, during input blank sample, standard specimen and sample to be tested are detected, control the temperature of thermal station Spend for 33~35 DEG C and keeping temperature it is constant.
- 9. Pb in water sample according to claim 82+Visible detection method, it is characterised in that inputted into detection chip empty When white sample, standard specimen or sample to be tested, control input flow is 200~8000 μ L/h, when indicator solution is inputted into detection chip, Control input flow is 50~2000 μ L/h.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610424594.8A CN106093328B (en) | 2016-06-14 | 2016-06-14 | A kind of Pb2+Pb in micro-fluidic detection chip and water sample2+Visible detection method |
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| CN201610424594.8A CN106093328B (en) | 2016-06-14 | 2016-06-14 | A kind of Pb2+Pb in micro-fluidic detection chip and water sample2+Visible detection method |
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