CN105102976A - Microfluidic chip for continuously monitoring chrome in water, and chrome detecting device comprising same - Google Patents

Microfluidic chip for continuously monitoring chrome in water, and chrome detecting device comprising same Download PDF

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CN105102976A
CN105102976A CN201380075382.8A CN201380075382A CN105102976A CN 105102976 A CN105102976 A CN 105102976A CN 201380075382 A CN201380075382 A CN 201380075382A CN 105102976 A CN105102976 A CN 105102976A
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reagent
chromium
sample
substrate
micro
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CN105102976B (en
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韩宗勋
安宰勋
缮井硲
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Academy Industry Foundation of POSTECH
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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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/76—Chemiluminescence; Bioluminescence
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F25/00—Flow mixers; Mixers for falling materials, e.g. solid particles
    • B01F25/40—Static mixers
    • B01F25/42—Static mixers in which the mixing is affected by moving the components jointly in changing directions, e.g. in tubes provided with baffles or obstructions
    • B01F25/43—Mixing tubes, e.g. wherein the material is moved in a radial or partly reversed direction
    • B01F25/433—Mixing tubes wherein the shape of the tube influences the mixing, e.g. mixing tubes with varying cross-section or provided with inwardly extending profiles
    • B01F25/4331—Mixers with bended, curved, coiled, wounded mixing tubes or comprising elements for bending the flow
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F33/00—Other mixers; Mixing plants; Combinations of mixers
    • B01F33/30—Micromixers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N31/00—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods
    • G01N31/22—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods using chemical indicators
    • 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
    • B01L3/502715—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 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces

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Abstract

A microfluidic chip according to one aspect of the present invention comprises: a first substrate formed so as to have a reduction channel for mixing samples and reagents, a reagent mixing channel for mixing reagents, and a detecting channel wherein the reduction channel and the reagent mixing channel are connected in parallel to an end part on one side thereof and an outlet is connected to an end part on the other side thereof; and a second substrate which has a detecting part formed facing the detecting channel such that light can pass therethrough and which is joined to the first substrate.

Description

For monitoring the micro-fluidic chip of chromium in water continuously and comprising its survey chromium device
Technical field
The present invention relates to a kind of micro-fluidic chip utilizing chemoluminescence method to monitor the chromium concn be present in water continuously and the survey chromium device comprising it.
Background technology
Chromium is the material being mainly used in alloy, pigment, leather, textile industry, catalyzer, wood preservative etc., owing to having the multiplex gilding preventing metal erosion of chemical stability.Along with the increase of above-mentioned industrial production activities, the discharge of chromium causes the pollution of surface water and underground water.
Natural chromium ion exists with multiple quantivalency form, but only there is the most stable chromium (III) and chromium (IV) in water.Chromium (III) is the essential nutrient participating in the metabolism such as glucose, cholesterol, fat, if lack chromium (III), body weight will reduce, and the function removing glucose from blood also can be caused impaired.And, if chromium (III) Long contact time skin, allergy or cancer may be caused.But chromium (III), to the solubleness of water and lower to biomembranous perviousness, therefore it is generally acknowledged that toxicity is not high.Unlike this, chromium (IV) to the solubleness of water and mobility higher, and oxidability and higher to biomembranous perviousness, brings harmful effect can to the various internal organs such as kidney, liver, lung, also can cause the inflammation of skin or mucous membrane.
As mentioned above, due to different qualities and the toxicity of chromium (III) and chromium (IV), analysis is carried out exactly to these respective concentration extremely important.Especially, chromium (III) and chromium (IV) are environmentally easily converted each other by redox reaction, therefore need to continue to monitor to water quality.According to the pollution discharge standard of Korea S; the total chromium concn be allowed in water resource conservation zone is 0.5ppm; then be restricted to below 2ppm in other areas, and the concentration of chromium (IV) is 0.1ppm in water resource conservation zone, is then restricted to below 0.5ppm in other areas.In addition, USEPA (EnvironmentalProtectionAgency, EPA) specifies that chromium (IV) is carcinogen, and the total chromium concn be allowed in potable water is limited in below 0.1ppm.
In the prior art for chromium in point bleed, utilize the chromium analysis of chemiluminescence reaction be measure Ru shown in following [reaction equation 1] in the basic conditions and chromium (III) catalyzer existence under by the luminol (luminol of hydrogen peroxide oxidation, 5-amino-2,3-dihydro-1,4-phthalazinedione) intensity of light (425nm) that sends.The concentration of the intensity of the light now sent and chromium (III) is proportional, and the intensity therefore by measuring light can carry out quantitative test to chromium (III).
[reaction equation 1]
For chromium (IV), because do not participate in the chemiluminescence reaction of luminol directly, so after using reductive agent that chromium (IV) is reduced into chromium (III) in acid condition, analyze total chromium concn.Chromium (IV) is reduced into the reaction of chromium (III) as shown in [reaction equation 2].
[reaction equation 2]
The method analyzed chromium (III) and chromium (IV) by chemoluminescence method is as follows: first, sample and reductive agent are reacted, after making the chromium in sample (IV) be reduced into chromium (III), then measure total chromium concn.Meanwhile, do not use reductive agent to measure the concentration of chromium (III), then the difference of concentration calculating total chromium and chromium (III) just can obtain the concentration of chromium (IV).Prior art is suitable for described method and have developed micro-fluidic chip for chromium in point bleed, but there are the following problems.
The first, in the basic conditions the luminol oxidation reaction caused by hydrogen peroxide hastens quickly due to reaction, in order to effectively measure maximum chemical luminescence, needs the length as far as possible reducing the chemical illuminating reagent hybrid channel injecting luminol and hydrogen peroxide.And, in order to analyze the concentration of chromium (IV) exactly, the chromium (IV) 100% in sample should be reduced into chromium (III), thus need the sufficient reaction time.Therefore, the length that should be designed to reduction passage is longer than the length of chemical illuminating reagent hybrid channel relatively.But, in the prior art, the length of chemical illuminating reagent hybrid channel is identical with for making chromium (IV) be reduced into the length of the reduction passage of chromium (III), therefore cannot detect the concentration of chromium (III) and chromium (IV) exactly, and detection sensitivity declines.
The second, utilize the chemiluminescence reaction of luminol not only to produce under the existence of chromium (III), also can produce under the existence of other metallic ions (iron (II), cobalt (II), copper (II), nickel (II) etc.).Prior art does not consider the effect to these interfering materials, therefore cannot the concentration of chromium in selectivity analytical sample.
Three, because the pH value of the process sample mixed with the distilled water injected when analyzing chromium (III) changes, there is the mixed crystallization that can generate chromium (III) and metallic ion in the on-the-spot sample of other metallic ions, and then the coprecipitation phenomena of chromium (III) also coprecipitation may occur.If generation coprecipitation phenomena, chromium (III) concentration of the sample injecting micro-fluidic chip will be caused to reduce, and then cause analytical error, described precipitation also can cause micro-fluidic chip blocked.
Four, in order to analyze chromium (III) and chromium (IV), two calibration curves (calibrationcurve, the calibration curve of total chromium and chromium (III)) are needed.
Five, enter detecting device because micro-fluidic chip transparent glass substrate makes light around, therefore detection sensitivity declines, and device also can be caused to use and become difficulty.
Summary of the invention
Technical matters
The invention provides a kind of in order to monitor chromium in water continuously by chemoluminescence method, detachment process need not be carried out can detect with high-level efficiency, high sensitivity chromium (III) and chromium (IV) under continuous flow state, and can when do not have interfering material to cause affect the micro-fluidic chip of selective enumeration method is carried out to chromium and comprises its survey chromium device.
Technical scheme
Micro-fluidic chip according to an aspect of the present invention, first substrate, is formed with the reduction passage for recombined sample and reagent, the reagent hybrid channel for mix reagent and a side end and is connected in parallel with described reduction passage and described reagent hybrid channel and the sense channel that end side is connected with escape hole; And second substrate, have towards described sense channel and make the test section of light therethrough, described second substrate is combined with described first substrate.
Described reduction passage more can be longer than described reagent hybrid channel, and described first substrate and second substrate can the material of color of extinction can be formed by having, or be dyed to can the color of extinction.
Described first substrate can be formed with the sample injection port for injecting sample and the reductive agent inlet for injecting reductive agent, described sample injection port take specimen access as mediator and reduction expanding channels, described reductive agent inlet take reducer channel as mediator and reduction expanding channels, and the first reagent inlet that described first substrate can be formed for injecting the first reagent and the second reagent inlet for injecting the second reagent, described first reagent inlet is connected with reagent hybrid channel by the first reagent passage, described second reagent inlet is connected with reagent hybrid channel by the second reagent passage.
Described sense channel can overlap and a side end is connected alternately with end side, described sense channel more more reduces to both sides side from central authorities, and described reduction passage can overlap with described reagent hybrid channel and a side end is connected alternately with end side.
Survey chromium device according to another aspect of the present invention, comprise: micro-fluidic chip, comprise first substrate and second substrate, wherein first substrate is formed with the reduction passage for recombined sample and reagent, the reagent hybrid channel for mix reagent and a side end and is connected in parallel with described reduction passage and described reagent hybrid channel and the sense channel that end side is connected with escape hole, and second substrate has towards described sense channel and make the test section of light therethrough, described second substrate is combined with described first substrate; Detecting device, is configured to towards described test section; And supply department, for supplying sample and reagent to described micro-fluidic chip.
Described reduction passage more can be longer than described reagent hybrid channel, and described first substrate and second substrate can the material of color of extinction can be formed by having, or be dyed to can the color of extinction.
Described first substrate can be formed the sample injection port for injecting sample, for inject reductive agent reductive agent inlet, for injecting the first reagent inlet of the first reagent and the second reagent inlet for injecting the second reagent.
Described supply department can to described sample injection port supply sample, to described reductive agent inlet supply potassium sulfite, be dissolved in the luminol of the buffer solution of alkali condition to described first reagent inlet supply, be dissolved in the hydrogen peroxide of the buffer solution of alkali condition to described second reagent inlet supply.
Bromide ion can be supplied to described micro-fluidic chip by described supply department together with described first reagent and described second reagent, and described supply department can by ethylenediamine tetraacetic acid (EDTA; Ethylenediaminetetraaceticacid) described micro-fluidic chip is supplied to together with described first reagent and described second reagent.
Described sense channel can overlap and a side end is connected alternately with end side, and described sense channel more more reduces to both sides side from central authorities.
Advantageous effects
In the present invention, make reduction passage in micro-fluidic chip and all can be reduced into chromium (III) to make the chromium in sample (IV), thus chromium (III) and chromium (IV) can be analyzed exactly.
And, the reduction passage of the reagent hybrid channel of injection chemical illuminating reagent and injection sample is made to have different length by asymmetrical design, use the chemiluminescent loss reducing as far as possible and produce in reagent hybrid channel, thus can low detection limit and high sensitivity chromium be detected.
And, in sense channel, be contained in the complexing agent of chemical illuminating reagent and sample meet and remove the metallic ion that the meeting existed in sample produces interference effect, thus under continuous flow state, selective enumeration method can be carried out to chromium.
And, inject sample by reductive agent inlet when analyzing chromium (III), therefore the precipitation of chromium (III) or the coprecipitation phenomena with interference metal can not occur, thus the analysis of chromium concn can be carried out exactly.
In addition, the slope 2 of the calibration curve of chromium (III), doubly to the slope of the calibration curve of total chromium, therefore only just can be analyzed the concentration of chromium (III) and chromium (IV), thus can shorten analysis time with the calibration curve of total chromium.
The micro-fluidic chip light absorbent of the present embodiment is made, in case light around enters into detecting device, thus can obtain highly sensitive stable chemiluminescence signal, easily be applied to scene.
Accompanying drawing explanation
Fig. 1 is the vertical view of the micro-fluidic chip upper plate that one embodiment of the invention is shown.
Fig. 2 is the vertical view of the micro-fluidic chip lower plate that one embodiment of the invention is shown.
Fig. 3 is the structural drawing of the survey chromium device that one embodiment of the invention is shown.
Fig. 4 a illustrates based on the chemiluminescence signal with or without 0.1M bromide ion and chromium (IV) concentration in chemical illuminating reagent.
Fig. 4 b illustrate based in chemical illuminating reagent with or without the calibration curve of the chromium (IV) of 0.1M bromide ion.
Fig. 5 illustrates the chemiluminescence signal based on the chromium obtained by black micro-fluidic chip and transparent micro-flow control chip (IV) concentration.
Fig. 6 is the photo observed chemiluminescence cooling type CCD (CooledCCD) camera of the 500ppb chromium (III) appearing at sense channel.
Fig. 7 a illustrates the chemiluminescence signal of the chromium (IV) based on reagent hybrid channel length variations.
Fig. 7 b illustrates the calibration curve of the chromium (IV) based on reagent hybrid channel length variations.
Fig. 7 illustrates chemiluminescence signal, these chemiluminescence signals are more longer than the micro-fluidic chip (micro-fluidic chip A) of reagent hybrid channel and the passage micro-fluidic chip (micro-fluidic chip B) identical with the length of reagent hybrid channel that reduce at reduction passage, measures with the standard sample that chromium (III) in the total chromium of 1000ppb is different with the ratio of chromium (IV).
Fig. 8 a illustrates the chemiluminescence signal of chromium (III) and chromium (IV) concentration obtained based on injecting reductive agent.
The calibration curve of the chromium (III) that Fig. 8 b illustrates injection reductive agent and obtains and chromium (IV).
Fig. 9 a illustrates the chemiluminescence signal based on total chromium and chromium (III) concentration.
Fig. 9 b illustrates the calibration curve of total chromium and chromium (III).
Figure 10 illustrates based on the chemiluminescence signal with or without EDTA and the chromium (III) with or without reductive agent in the sample being mixed with chromium (III) and iron (II).
Figure 11 illustrates and injects distilled water in the sample being mixed with chromium (III) and iron (II) and the chemiluminescence signal of the chromium (III) obtained.
Embodiment
Below, embodiments of the present invention will be described in detail with reference to the accompanying drawings, easily implements the present invention to make those skilled in the art in the invention.The present invention can implement in a variety of different ways, is not limited to following embodiment.
Fig. 1 is the vertical view of the micro-fluidic chip upper plate that one embodiment of the invention is shown, Fig. 2 is the vertical view of the micro-fluidic chip lower plate that one embodiment of the invention is shown.
The micro-fluidic chip 30 of the present embodiment comprises upper plate (first substrate) 10 and lower plate (second substrate) 20.Upper plate 10 side of being plate shape, upper plate 10 is formed with sample injection port 11, reductive agent inlet 12, first reagent inlet 13 and the second reagent inlet 14.And, sample injection port 11 being formed with the linking part 31 for being combined with sample ascending pipe 52, reductive agent inlet 12 being formed with the linking part 32 for being combined with reductive agent ascending pipe 53.
First reagent inlet 13 is formed with the linking part 35 linking part 34, the second reagent inlet 14 for being combined with reagent ascending pipe 54 is formed with for being combined with reagent ascending pipe 55.
In addition, upper plate 10 is formed with specimen access 11a, reducer channel 12a, the first reagent passage 13a, the second reagent passage 14a, reduction passage 15, reagent hybrid channel 16 and sense channel 17.Sample injection port 11 is connected with specimen access 11a, and reductive agent inlet 12 is connected with reducer channel 12a.Injecting containing the sample of chromium by sample injection port 11, being injected for reducing the reductive agent such as potassium sulfite of chromium (IV) by reductive agent inlet 12.
Specimen access 11a and reducer channel 12a are connected with reduction passage 15, in reduction passage 15 sample and reductive agent mixed, the chromium (IV) comprised in the sample is reduced agent and is reduced into chromium (III).Reduction passage 15 overlaps and a side end is connected alternately with end side.One side end of reduction passage 15 is connected with specimen access 11a and reducer channel 12a, and end side is connected with sense channel 17.
First reagent inlet 13 is connected with the first reagent passage 13a, and the second reagent inlet 14 is connected with the second reagent passage 14a.Can being injected by the first reagent inlet 13 and be dissolved in the luminol of alkali condition buffer solution, bromide ion and EDTA, the hydrogen peroxide of the alkali condition buffer solution be dissolved in for dissolving described luminol, bromide ion and EDTA can being injected by the second reagent inlet 14.
First reagent passage 13a and the second reagent passage 14a is connected with reagent hybrid channel 16, in reagent hybrid channel 16 luminol and hydrogen peroxide mixed.Reagent hybrid channel 16 overlaps and a side end is connected alternately with end side.One side end of reagent hybrid channel 16 is connected with the first reagent passage 13a and the second reagent passage 14a, and end side is connected with sense channel 17.
One side end of sense channel 17 is connected with reduction passage 15 and reagent hybrid channel 16, and end side is formed with escape hole 18, so that the reagent of discharging for reacting and sample.Escape hole 18 is formed with linking part 33, to be combined with vent pipe.
Sense channel 17 overlaps and a side end is connected alternately with end side.The length of overlapping sense channel 17 more more reduces to both sides side from central authorities, and the center configuration of sense channel 17 becomes to align with the center in circle detection portion 21.
One side end of sense channel 17 is connected in parallel with reduction passage 15 and reagent hybrid channel 16, the sample therefore met in sense channel and reagent mixed, thus eliminate the interference effect in sample, produce chemiluminescence reaction simultaneously.
The length of reduction passage 15 is 2 times of reagent hybrid channel 16 length to 6 times.Thus, chromium (IV) can all be reduced agent through reduction passage 15 and be reduced into chromium (III).And the length due to reagent hybrid channel 16 is formed shorter, make the chemiluminescent loss occurred in reagent hybrid channel 16 reduce to minimum, thus can detect chromium with low detection limit and high sensitivity.
Linking part 31,32,33,34,35 is formed by polymkeric substance or pipe (tubing) etc., and has hole so that pipe or pipeline combine with can being easy to handling.
Upper plate 10 can be formed by the polymkeric substance with light absorptive, particularly can be formed by black dimethyl silicone polymer (PDMS, polydimethylsiloxane).But upper plate 10 can be formed by the material of the shades of colour with light absorptive, or can be dyed to the color with light absorptive.
In addition, corresponding with the sense channel 17 of upper plate 10 in lower plate 20 bottom is formed with test section 21.Lower plate 20 is formed by the PDMS of black, and test section 21 is filled with transparent PDMS so that the light therethrough produced by chemiluminescence reaction.Test section 21 is formed by the mode injecting transparent PDMS after black PDMS forms hole.But, when lower plate 20 is colored, only have test section to be colored and to be formed as transparent.
As mentioned above, when upper plate 10 and lower plate 20 are formed as having light absorptive, the light injected from outside will be absorbed, thus stable chemiluminescence signal can be obtained, and detection limit and sensitivity can be improved.
Below, the manufacture method of upper plate 10 and lower plate 20 is described.On ready silicon wafer after spin coating photoresist, irradiation ultraviolet radiation under the state of placing photomask, then form positive needle drawing case through solidification and development.
Upper plate 10, after solidification, just can be obtained from wafer-separate the black PDMS upper plate being formed with cloudy needle drawing case by the silicon wafer upper black PDMS being formed with positive needle drawing case.Lower plate 20 also adopts the method manufacture identical with upper plate 10.
For upper plate 10 and lower plate 20, after its bound fraction carries out surface treatment by arc discharge, combine in the Centered mode of the test section 21 of the center of the sense channel 17 of upper plate 10 and lower plate 20.
Sample and reductive agent move to sense channel 17 through reduction passage 15, and reagent moves to sense channel 17 through reagent hybrid channel 16, thus causes chemiluminescence reaction.Sample after sense channel 17 causes chemiluminescence reaction and reagent move to escape hole 18.
Fig. 3 is the structural drawing of the survey chromium device that one embodiment of the invention is shown.
Known with reference to Fig. 3, the survey chromium device of the present embodiment, to comprise in the housing 41 that is inserted in and forms profile and to be configured in detecting device 42 below test section 21, micro-fluidic chip 30 and to supply the supply department 51 of sample and reagent to micro-fluidic chip 30.Housing 41 is formed as the boxed-section shape slightly in rectangular parallelepiped.Housing 41 is formed by the metal dying black to absorb light, and micro-fluidic chip 30 is fixedly arranged on above housing 41.Thus, block extraneous light and enter detecting device 42, and then only have the light produced through chemiluminescence reaction to inject detecting device 42 by test section 21.
In addition, the linking part 31 being located at sample injection port 11 is equipped with sample ascending pipe 52, the linking part 32 being located at reductive agent inlet 12 is equipped with reductive agent ascending pipe 53.And, the linking part 34 being located at the first reagent inlet 13 is equipped with the first reagent ascending pipe 54, the linking part 35 being located at the first reagent inlet 14 is equipped with the second reagent ascending pipe 55.
Supply department 51 is provided with peristaltic pump, and supply department 51 supplies sample to sample ascending pipe 52, and supplies potassium sulfite to reductive agent ascending pipe 53.And supply department 51 supplies luminol to the first reagent ascending pipe 54, and supply hydrogen peroxide to the second reagent ascending pipe 55.
Supply department 51 reagent without interruption and sample, be discharged by escape hole 18 after the sample supplied and reagent reacting.Thus, the survey chromium device of the present embodiment can monitor chromium continuously.
Bromide ion is fed to micro-fluidic chip 30 by supply department 51 together with reagent, and chemiluminescence intensity increases thus, and then improves the sensitivity of surveying chromium device, surveys chromium device and can have lower detection limit.And EDTA supplies by supply department 51 together with reagent.EDTA reacts with metallic ion in the basic conditions and forms complex compound, and removing participates in other metallic ions of the chemiluminescence reaction of luminol contained by sample thus, thus the mensuration of chromium concn can be avoided inaccurate.
Detecting device 42 accesses power supply 61.And detecting device 42 connects the digital multimeter 62 of establishing and carrying out processing process to the signal produced in detecting device 42, the signal after process is sent to connected computing machine 63 by digital multimeter 62.
[experimental example 1]
In experimental example 1, the micro-fluidic chip utilizing the present embodiment and the survey chromium device comprising it determine the chemiluminescence intensity based on chromium concn.Chemical illuminating reagent is used for detecting chromium (III) from sample, and it carries out reaction with chromium (III) in the basic conditions and causes chemiluminescence.Chemical illuminating reagent is made up of luminol and hydrogen peroxide, and chemiluminescent intensity can change according to the concentration of luminol and hydrogen peroxide.
Buffer solution preparation method for chemoluminescence method is as follows: by the boric acid (H of 30.09g 3bO 3), the potassium bromide (KBr) of 10.2g, the EDTA ((HOOCCH of 2.92g 2) 2hNCH 2cH 2nH (CH 2cOOH) 2) be dissolved in distilled water, potassium hydroxide (KOH) solution of recycling 5M by pH value furnishing 10.9, and makes final volume become 1L.The preparation method of chemical illuminating reagent is as follows: the luminol of 0.02g is dissolved in buffer solution and after adjusted to ph reaches 10.9, makes final volume become 100mL.The preparation method of oxygenant is as follows: the hydrogen peroxide of 0.868mL is dissolved in described buffer solution and after adjusted to ph reaches 10.9, makes final volume become 100mL.As follows for making chromium (IV) be reduced into the reductive agent its preparation method of chromium (III): by the potassium sulfite (K of 0.1104g 2sO 3) be dissolved in distilled water and after adjusted to ph reaches 2.5, make final volume become 100mL.The standard sample its preparation method of chromium (III) and chromium (IV) is as follows: respectively by chromic nitrate nonahydrate (Cr (NO 3) 39H 2and potassium chromate (K O) 2crO 4) be dissolved in after distilled water prepares the solution of 1000ppm, be prepared respectively with the concentration of 5ppb, 50ppb, 125ppb, 250ppb, 500ppb, 1000ppb by dilution.
Chemical illuminating reagent and luminol are injected by the first reagent inlet 13, and hydrogen peroxide is injected by the second reagent inlet 14.Sample is injected by reductive agent inlet 12 respectively by sample injection port 11, reductive agent.The flow velocity of the peristaltic pump used when injecting each solution is 7.5 μ L/min.
Fig. 4 a illustrates based on the chemiluminescence signal with or without 0.1M bromide ion and chromium (IV) concentration in chemical illuminating reagent, Fig. 4 b illustrate based in chemical illuminating reagent with or without the calibration curve of the chromium (IV) of 0.1M bromide ion.
The chromium (IV) that the standard sample used in this experimental example is 5ppb, 50ppb, 125ppb, 250ppb, 500ppb, 1000ppb, inject reductive agent and use the chemical illuminating reagent adding 0.1M bromide ion and the chemical illuminating reagent not adding bromide ion, METHOD FOR CONTINUOUS DETERMINATION is based on the chemiluminescence signal of each concentration.Under bromide ion can attack alkali condition, metallic ion, hydrogen peroxide and luminol react and the complex of generation.Thus, can generate and more cause chemiluminescent excited state O-phthalic amine (3-aminophthalate), chemiluminescence intensity is increased.As shown in fig. 4 a, when using the chemical illuminating reagent adding 0.1M bromide ion, compared with when not adding the chemical illuminating reagent of bromide ion with use, chemiluminescence intensity greatly increases.
In fig. 4b, the calibration curve adding the chemical illuminating reagent of 0.1M bromide ion is y=0.00100x+0.00289 (R 2=0.9999) calibration curve, not adding the chemical illuminating reagent of bromide ion is y=0.000182x+0.000212 (R 2=0.9999).Wherein, y represents chemiluminescence intensity, and x represents concentration, R 2represent linear coefficient.This calibration curve uses the standard sample prepared by material to be analyzed, according to after concentration sequential determination chemiluminescence intensity from low concentration, the relation of the chemiluminescence intensity based on material concentration to be analyzed represented with line.
As shown in Figure 4 b, when using the chemical illuminating reagent adding 0.1M bromide ion, compared with when not adding the chemical illuminating reagent of bromide ion with use, the slope increase of calibration curve about 5.5 times.
And when adding 0.1M bromide ion, detection limit is 0.32ppb, compared with detection limit 1.3ppb when not adding bromide ion, has the detection limit of approximately low 4 times.Therefore, can detect chromium with higher sensitivity and low detection limit by adding 0.1M bromide ion in chemical illuminating reagent.
[experimental example 2]
In this experimental example 2, upper plate 10 and lower plate 20 are all made of black PDMS and be applicable to the micro-fluidic chip that transparent PDMS makes survey chromium device, and determines the chemiluminescence intensity of total chromium.This experimental example carries out in darkroom.
Fig. 5 illustrates the chemiluminescence signal based on the chromium obtained by black micro-fluidic chip and transparent micro-flow control chip (IV) concentration.
As shown in Figure 5, the micro-fluidic chip made of transparent PDMS enters detecting device due to extraneous light, the intensity of background signal (backgroundsignal) and the noise of signal greatly increase, and the chemiluminescence signal obtained by chromium (IV) standard sample (5ppb, 50ppb, 125ppb, 250ppb, 500ppb, 1000ppb) also causes jitter because of the scattering of light that produces in the light of surrounding and chemiluminescence reaction and noise is large.
And along with background signal intensity increases greatly, the chemiluminescence signal of the chromium (III) of below 50ppb cannot be distinguished with background signal.As mentioned above, for the chemiluminescence signal obtained by the micro-fluidic chip using transparent PDMS to make, along with the intensity of background signal and the increase of noise, detection limit uprises, and the concentration range of detectable chromium reduces.
Review the micro-fluidic chip using black PDMS to make, enter detecting device owing to having blocked extraneous light, the intensity of background signal and noise reduction, chemiluminescence signal is highly stable.Survey chromium device and effectively blocked extraneous light by using the micro-fluidic chip made by black PDMS, thus can detect chromium with higher sensitivity and low detection limit, the concentration range of detectable chromium can also be expanded.
[experimental example 3]
In this experimental example 3,500ppb chromium (III) is injected by sample injection port, and observe chemiluminescence with cooling type CCD (CooledCCD) camera.Fig. 6 is the photo observed chemiluminescence cooling type CCD (CooledCCD) camera of the 500ppb chromium (III) appearing at sense channel.Chemiluminescence occurs the strongest at the center section of sense channel 17, and start-up portion is relative with the chemiluminescence intensity of end part is weaker than center section.This result shows, the design of micro-fluidic chip and the condition of reagent are optimized, and makes it possible to measure the chemically luminous most of light occuring to disappearance and produced by chemiluminescence reaction, thus can carry out highly sensitive chromium detection.
[experimental example 4]
In this experimental example 4, the length of reduction passage is fixed into certain length 100cm, and the length changing reagent hybrid channel 16 is respectively 0cm, 20cm, 60cm, 100cm, and determines chemiluminescence intensity.
Fig. 7 a illustrates the chemiluminescence signal of the chromium (IV) based on reagent hybrid channel length variations, and Fig. 7 b illustrates the calibration curve of the chromium (IV) based on reagent hybrid channel length variations.
The chromium (IV) that the standard sample used in this experimental example is 5ppb, 50ppb, 125ppb, 250ppb, 500ppb, 1000ppb, injects the chemiluminescence signal of reductive agent METHOD FOR CONTINUOUS DETERMINATION based on each concentration.The reaction velocity of the luminol oxidation reaction caused by hydrogen peroxide under alkali condition quickly, even if therefore chemical illuminating reagent and luminol and hydrogen peroxide be not when playing chromium (III) of catalytic action, also carry out reacting in reagent hybrid channel 16 and causing chemiluminescence.Therefore, as shown in Figure 7a, the length of reagent hybrid channel 16 is longer, and the chemiluminescent intensity occurred at sense channel 17 more reduces, and Fig. 7 b is the calibration curve obtained by Fig. 7 a, and along with the length of reagent hybrid channel 16, the slope of calibration curve reduces.But, if there is no reagent hybrid channel 16, when reagent directly being injected sense channel (reagent hybrid channel is 0cm), because produce large pressure differential with the missionary society of the length of reduction passage 15, cause chemiluminescence signal very unstable, on the calibration curve of Fig. 7 b, the chemiluminescence intensity of 500ppb chromium (IV) exceeds the range of linearity.
The slope of calibration curve and the reappearance of mensuration are depended in the sensitivity of pick-up unit.In fig .7b, the length except reagent hybrid channel 16 is except the situation of 0cm, can suppose that the reappearance measured is identical for other situations.Therefore, the chromium detection sensitivity based on reagent hybrid channel 16 length depends on the slope of calibration curve, so the highest when the length of reagent hybrid channel 16 is 20cm.
[experimental example 4]
In this experimental example 4, in order to confirm that chromium (IV) is reduced into the percent reduction of chromium (III), prepare total chromium concn be all 1000ppb and chromium (III) different with the ratio of chromium (IV) be respectively 3: 7,5: 5,7: 3 standard sample, this standard sample is injected micro-fluidic chip A and micro-fluidic chip B respectively successively, and determines chemiluminescence signal.
Fig. 7 illustrates chemiluminescence signal, these chemiluminescence signals are more longer than the long micro-fluidic chip A in reagent hybrid channel (reduction passage 15:100cm at reduction passage, reagent hybrid channel 16:20cm) with the reduction passage micro-fluidic chip B identical with the length of reagent hybrid channel (reduction passage 15:20cm, reagent hybrid channel 16:20cm), measure with standard sample that chromium (III) in the total chromium of 1000ppb is different with the ratio of chromium (IV).
For micro-fluidic chip A, the chromium (IV) comprised in sample is all reduced into chromium (III), although the composition of chromium (III) in sample and chromium (IV) is than different, demonstrates identical chemiluminescence intensity.
For micro-fluidic chip B, only some is reduced into chromium (III) to the chromium (IV) comprised in sample, and the sample therefore with heterogeneity ratio demonstrates different chemiluminescence intensities respectively.
As mentioned above, according to the present invention, because the length of reduction passage 15 is longer than the length of reagent hybrid channel 16, the concentration of chromium (IV) can be measured exactly.
[experimental example 5]
In this experimental example 5, in order to confirm that whether chromium (IV) that concentration range is large 100% be reduced into chromium (III), the chromium (III) of 5ppb, 50ppb, 125ppb, 250ppb, 500ppb, 1000ppb and chromium (IV) standard sample are injected sample injection port 11 respectively successively, reductive agent is injected reductive agent inlet 12, and determine chemiluminescence signal.
Fig. 8 a illustrates and injects reductive agent and the chemiluminescence signal based on chromium (III) and chromium (IV) concentration that obtains, the calibration curve of the chromium (III) that Fig. 8 b illustrates injection reductive agent and obtains and chromium (IV).
As shown in Figure 8 a, the chromium (III) of each concentration and chromium (IV) demonstrate almost identical chemiluminescence intensity.
In figure 8b, the calibration curve of chromium (III) is y=0.00100x-0.00115 (R 2=0.9999), the calibration curve of chromium (IV) is y=0.00100x+0.00127 (R 2=0.9999).Wherein, y represents chemiluminescence intensity, and x represents concentration, R 2represent linear coefficient.
Chromium (III) is identical with its slope of calibration curve of chromium (IV), and y-axis intercept has very small difference, therefore respective calibration curve is shown in the result on a figure, as shown in Figure 8 b two calibration curve overlaps.
From the result of Fig. 8, chromium (IV) 100% large for concentration range (5 ~ 1000ppb) can be reduced into chromium (III) by micro-fluidic chip and the chemiluminescence detecting comprising it, therefore can detect the concentration of chromium (III) and chromium (IV) exactly, not have analytical error.
[experimental example 6]
This experimental example 6 determines the chemiluminescence signal based on concentration obtained by total chromium and chromium (III) standard sample of 5ppb, 50ppb, 125ppb, 250ppb, 500ppb, 1000ppb respectively.
Fig. 9 a illustrates the chemiluminescence signal based on total chromium and chromium (III) concentration, and Fig. 9 b illustrates the calibration curve of total chromium and chromium (III).
For the sample of same concentrations, the chemiluminescence intensity of chromium (III) is 2 times of total chromium chemiluminescence intensity.As shown in Figure 8 a, when injecting reductive agent, chromium (III) and the chromium (IV) of same concentrations have almost similar chemiluminescence intensity.But, in this experimental example 6, when analyzing chromium (III), sample is injected by sample injection port 11 and reductive agent inlet 12, and during analyzing total chromium, only inject sample by sample injection port 11, and inject reductive agent by reductive agent inlet 12, therefore the concentration of sample is diluted as 1/2, so the chemiluminescence intensity of the chromium of same concentrations (III) is 2 times of the chemiluminescence intensity of total chromium.
Fig. 9 b obtains calibration curve for total chromium and chromium (III) by shown in Fig. 9 a respectively in triplicate based on the chemiluminescence signal of concentration, and known reappearance is very high, and thus relative standard deviation is very little.In figure 9b, the calibration curve of chromium (III) is y=0.00200x+0.00112 (R 2=0.9999), the calibration curve of total chromium is y=0.00100x+0.00289 (R 2=0.9999).Wherein, y represents chemiluminescence intensity, and x represents concentration, R 2represent linear coefficient.
According to this experimental example 6, the slope of chromium (III) calibration curve is 2 times of the slope of total chromium calibration curve.Utilize this relation, analyze total chromium of existing in on-the-spot sample and chromium (III), the result of concentration of chromium (IV) and the result similar (table 2) calculated by two calibration curves (calibration curve of total chromium and chromium (III)) by total chromium calibration curve.As mentioned above, survey chromium device of the present invention can analyzing total chromium and chromium (III), chromium (IV) exactly by means of only total chromium calibration curve, can greatly reduce analysis time.
The total chromium obtained from this experimental example 6 and the respective detection limit of chromium (III) are 0.32ppb, 0.13ppb.And linearity range is that 5ppb ~ 1000ppb comprises legal restrictions concentration, dilution or the pre-service such as concentrated that therefore need not carry out sample just can carry out chromium detection.
[experimental example 7]
Luminol chemiluminescence is reacted, except chromium (III), other metallic ions (cobalt (II), iron (II), copper (II), nickel (II) etc.) also participate in reaction, and metallic ion comprises chromium and relies on pH value to a great extent to the solubleness of water and mobility.Such as, the reasons such as acid rain can cause the acidifying of water, make the pH value of water be down to 4.0 ~ 4.5 and cause the concentration of metallic ion to increase, and the impact that interference metallic ion causes can become more serious.The object of this experimental example 7 confirms, when the acidifying of water causes pH value to change, whether can also carry out selective enumeration method when the interference effect not having other metallic ions to cause to chromium.
For the effect that interference metallic ion causes, use EDTA just can eliminate.EDTA reacts with metallic ion in the basic conditions and forms complex compound, and the metallic ion forming complex compound can not participate in chemiluminescence.On the contrary, form complex compound compared with other metallic ions with relatively slow speed when chromium (III) and EDTA react, be therefore applicable to the selectivity analysis to chromium in sample.
Figure 10 illustrates based on the chemiluminescence signal with or without EDTA and the chromium (III) with or without reductive agent in the sample being mixed with chromium (III) and iron (II), in Fig. 10, a () is by the chemiluminescence signal of the Specimen Determination being only dissolved with 500ppb chromium (III), as the benchmark of the chemiluminescence intensity of other samples, the chemiluminescence intensity of (a) is set as 100%.
In Fig. 10, (b) be the sample being dissolved with 500ppb chromium (III) and 5ppm iron (II) is used do not add the luminol of EDTA, chemiluminescence intensity that hydrogen peroxide is analyzed, demonstrate the chemiluminescence intensity of 117%.It is produce because disturbing material and 5ppm iron (II) that chemiluminescence intensity increases by 17%.
In Fig. 10, c () injects the sample identical with (b) and the reagent that injection is dissolved with luminol, the buffer solution of hydrogen peroxide dissolves the EDTA of 10mM respectively carries out the chemiluminescence signal that measures, demonstrate the chemiluminescence intensity of identical with (a) 100%.From described result, use the EDTA of 10mM just can carry out selective enumeration method when the interference effect not having 5ppm iron (II) to cause to chromium (III).
In Fig. 10, (d) is the chemiluminescence signal injecting reductive agent by an inlet in two sample injection port and obtain under the condition identical with (c), demonstrates the chemiluminescence intensity being equivalent to 50%.The chemiluminescence intensity of 50% is because the injection of reductive agent causes the concentration of sample to be diluted as 1/2.From described result, when using the EDTA of 10mM, even if the reductive agent that injection pH value is 2.5 also can not have an impact to chemiluminescence signal.
From the result of Figure 10, even if when acidifying causes disturbing the concentration of metallic ion to uprise, detachment process need not be carried out and uses EDTA also can carry out selective enumeration method to chromium (III) and chromium (IV).
[experimental example 8]
This experimental example 8 in order to injection distilled water time confirmation analysis chromium (III) whether produces sediment carries out, the solution ph furnishing 4.5 being dissolved with 500ppb chromium (III) and 5ppm iron (II) is got out sample, this sample is injected by sample injection port 11 and reductive agent inlet 12, confirm chemiluminescence signal now, and be dissolved with the solution of 500ppb chromium (III) and iron (II) by sample injection port 11 injection and injected distilled water by reductive agent inlet 12, confirm chemiluminescence signal now.
Distilled water (pH5.8 ~ 6.2) is filled with by an inlet in two inlets during the chromium of prior art in test samples (III).For described situation, because the pH value of sample changes in reduction passage 20 sample and the mixed process of distilled water, the precipitation of chromium (III) may be caused.
Figure 11 illustrates and injects distilled water in the sample being mixed with chromium (III) and iron (II) and the chemiluminescence signal obtaining chromium (III).In fig. 11, (a) is the chemiluminescence signal that 500ppb chromium (III) is injected by sample injection port 11 and reductive agent inlet 12 and obtained, and its chemiluminescence intensity is set as 100%.In fig. 11, b () is the chemiluminescence signal that the sample of the 500ppb chromium (III) and 5ppm iron (II) that are dissolved with pH4.5 is injected respectively by sample injection port 11 and reductive agent inlet 12 and obtained, show chemiluminescence intensity ratio (a) high by 17% in Figure 11.In fig. 11, (d) is only had to be the chemiluminescence signal obtained by adding EDTA in reagent.Therefore, in (b) of Figure 11, chemiluminescence intensity increases by 17% and is interference effect because 5ppm iron (II) causes and produces.
In fig. 11, (c) injects the sample being dissolved with 500ppb chromium (III) and 5ppm iron (II) and the chemiluminescence signal injecting distilled water by reductive agent inlet and obtain by sample injection port.If sample is not affected because of the injection of distilled water, then because the concentration of sample is diluted as 1/2, the chemiluminescence intensity of 58.5% should be shown.But in fig. 11, (c) demonstrates the chemiluminescence intensity of 17.3%.This shows that the sample of pH4.5 in reduction passage mixes rear pH value and uprises with distilled water, and then produces the deposited phenomenon of chromium (III) or the coprecipitation phenomena of iron and chromium, causes chemiluminescence intensity to reduce 41.2%.In fig. 11, d () is by the method identical with (c) injection sample and distilled water and injects the reagent being dissolved with EDTA and the chemiluminescence signal obtained, represent in sample and removed by EDTA with the iron (II) that dissolved state is residual, cause the chemiluminescence intensity of 5.4% to reduce.From described result, when detecting chromium (III) by chemoluminescence method, if inject distilled water analytical error will occur, even if injection EDTA also cannot eliminate the interference effect that other metallic ions cause in this case.
[experimental example 9]
In this experimental example 9, use the reagent not adding EDTA and the reagent of EDTA adding 10mM, chemiluminescence detection has been carried out to the sample participating in the iron (II) of luminol chemiluminescence reaction, cobalt (II), copper (II), nickel (II) and the pH4.5 that is mixed with chromium.
The chromium (III) and described interference metallic ion that are dissolved with 500ppb are dissolved with respectively to the sample of 500ppb or 5ppm, utilize nitric acid by pH value furnishing 4.5.Now, the volume for the salpeter solution of adjusted to ph is less than 1/1000 of volume of sample, can ignore the concentration change of chromium and interference metal in sample.And, the total chromium (comprising the chromium (III) of 250ppb and the chromium (IV) of 250ppb) and interference metallic ion that are dissolved with 500ppb are dissolved with respectively to the sample of 500ppb or 5ppm, utilize nitric acid by pH value furnishing 4.5.
Chemical illuminating reagent be EDTA by adding 10mM buffer solution and do not add EDTA buffer solution in add luminol respectively and prepared by hydrogen peroxide.
[table 1]
[table 1] illustrates the chemiluminescence intensity of each sample, it is the sample for the chromium being dissolved with 500ppb (III) and interference metallic ion, the chemiluminescence intensity of 500ppb chromium (III) sample is set as 100%, for the sample being dissolved with the total chromium of 500ppb and interference metallic ion, the chemiluminescence intensity of total chromium sample of 500ppb is set as 100%, and when injecting the chemical illuminating reagent of the chemical illuminating reagent not adding EDTA and the EDTA adding 10mM, for the chemiluminescence intensity of each sample.Value on the right side of chemiluminescence intensity in bracket is the relative standard deviation measuring chemiluminescence intensity three times for each sample and obtain.
When inject do not add the reagent of EDTA time, the chemiluminescence intensity measured due to the interference metallic ion existed in sample is more than 100%.But, when using the chemical illuminating reagent of the EDTA adding 10mM, in sense channel, disturb metallic ion to be removed by EDTA, thus total chromium standard sample of the chromium (III) of chemiluminescence intensity and 500ppb, 500ppb is identical, concentration for interference metallic ion is the sample of 10 times (5ppm) of chromium concn, also can carry out selectivity analysis when not having interference effect to the chromium (III) of 500ppb and total chromium.
[experimental example 10]
This experimental example 10, in order to confirm that the performance of the pick-up unit of the present embodiment is carried out, analyzes the concentration of total chromium and chromium (III) in the sample collected in factory, chromium (IV).The sample of the metre filter through having 0.45 μm of pore is injected micro-fluidic chip, and determines chromium concn.
Table 2 is the forms analysis result of chromium total in sample and chromium (III), chromium (IV) concentration and the result by atomic absorption and light absorption method analysis being carried out contrasting, atomic absorption can only analyzing total chromium concn, and detection limit is 20ppb.And, light absorption method adopts UV-visible spectrophotometric determination by chromium (IV) and 1,5-diphenylcarbazide (1, the absorbance log of the red complex that reaction 5-diphenylcarbazide) generates under 540nm wavelength is also analyzed, due to the interference effect not having other materials to cause, be the analytical approach that selectivity is high in chromium (IV) is analyzed.
To according to the chromium in the sample of this experimental example 10, the micro-fluidic chip of the present embodiment is used to be carried out the result analyzed by chemoluminescence method, the concentration of the total chromium comprised in sample 1, sample 2, sample 3 is respectively 137ppb, 112ppb, 128ppb, be similar to the result analyzed by atomic absorption, the relative standard deviation lower than atomic absorption is obtained to the result of each sample analysis three times.
[table 2]
For the concentration of the chromium analyzed by chemoluminescence method (III), sample 1, sample 2, sample 3 are respectively 27ppb, 20ppb, 28ppb, are similar to and deduct chromium (IV) concentration analyzed by light absorption method and chromium (III) concentration calculated from the total chromium concn analyzed by atomic absorption.For the concentration of chromium (IV), utilize the calculated value of total chromium and chromium (III) concentration similar with chromium (IV) concentration analyzed by light absorption method.As mentioned above, utilize the survey chromium device analysis of the present embodiment, the concentration of total chromium and chromium (III), chromium (IV) can be measured exactly.
Be described above preferred embodiment of the present invention, but the present invention is not limited to foregoing, can various mode of texturing implement within the scope of claims and instructions and accompanying drawing thereof, undoubtedly these all belong within protection scope of the present invention.

Claims (15)

1. a micro-fluidic chip, comprising:
First substrate, is formed with the reduction passage for recombined sample and reagent, the reagent hybrid channel for mix reagent and a side end and is connected in parallel with described reduction passage and described reagent hybrid channel and the sense channel that end side is connected with escape hole; And
Second substrate, have towards described sense channel and make the test section of light therethrough, described second substrate is combined with described first substrate.
2. micro-fluidic chip according to claim 1, wherein,
Described reduction passage is more longer than described reagent hybrid channel.
3. micro-fluidic chip according to claim 2, wherein,
Described first substrate and second substrate can the material of color of extinction be formed by having, or be dyed to can the color of extinction.
4. micro-fluidic chip according to claim 2, wherein,
Described first substrate is formed with the sample injection port for injecting sample and the reductive agent inlet for injecting reductive agent, described sample injection port take specimen access as mediator and reduction expanding channels, and described reductive agent inlet take reducer channel as mediator and reduction expanding channels.
5. micro-fluidic chip according to claim 4, wherein,
Described first substrate is formed with the first reagent inlet for injecting the first reagent and the second reagent inlet for injecting the second reagent, described first reagent inlet is connected with reagent hybrid channel by the first reagent passage, and described second reagent inlet is connected with reagent hybrid channel by the second reagent passage.
6. micro-fluidic chip according to claim 2, wherein,
Described sense channel overlaps and a side end is connected alternately with end side, and described sense channel more more reduces to both sides side from central authorities.
7. micro-fluidic chip according to claim 2, wherein,
Described reduction passage overlaps with described reagent hybrid channel and a side end is connected alternately with end side.
8. survey a chromium device, comprising:
Micro-fluidic chip, comprise first substrate and second substrate, wherein first substrate is formed with the reduction passage for recombined sample and reagent, the reagent hybrid channel for mix reagent and a side end and is connected in parallel with described reduction passage and described reagent hybrid channel and the sense channel that end side is connected with escape hole, and second substrate has towards described sense channel and make the test section of light therethrough, described second substrate is combined with described first substrate;
Detecting device, is configured to towards described test section; And
Supply department, for supplying sample and reagent to described micro-fluidic chip.
9. survey chromium device according to claim 8, wherein,
Described reduction passage is more longer than described reagent hybrid channel.
10. survey chromium device according to claim 9, wherein,
Described first substrate and second substrate can the material of color of extinction be formed by having, or be dyed to can the color of extinction.
11. survey chromium devices according to claim 9, wherein,
Described first substrate is formed the sample injection port for injecting sample, for inject reductive agent reductive agent inlet, for injecting the first reagent inlet of the first reagent and the second reagent inlet for injecting the second reagent.
12. survey chromium devices according to claim 11, wherein,
Described supply department is to described sample injection port supply sample, to described reductive agent inlet supply potassium sulfite, be dissolved in the luminol of the buffer solution of alkali condition to described first reagent inlet supply, be dissolved in the hydrogen peroxide of the buffer solution of alkali condition to described second reagent inlet supply.
13. survey chromium devices according to claim 12, wherein,
Bromide ion is supplied to described micro-fluidic chip by described supply department together with described first reagent and described second reagent.
14. survey chromium devices according to claim 12, wherein,
Ethylenediamine tetraacetic acid is supplied to described micro-fluidic chip by described supply department together with described first reagent and described second reagent.
15. survey chromium devices according to claim 9, wherein,
Described sense channel overlaps and a side end is connected alternately with end side, and described sense channel more more reduces to both sides side from central authorities.
CN201380075382.8A 2013-04-05 2013-11-12 Micro-fluidic chip for chromium in continuous monitoring water and the survey chromium device comprising it Expired - Fee Related CN105102976B (en)

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