CN108760730A - A kind of method of paper substrate double mode detection magnesium ion - Google Patents

A kind of method of paper substrate double mode detection magnesium ion Download PDF

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CN108760730A
CN108760730A CN201810453724.XA CN201810453724A CN108760730A CN 108760730 A CN108760730 A CN 108760730A CN 201810453724 A CN201810453724 A CN 201810453724A CN 108760730 A CN108760730 A CN 108760730A
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solution
paper
dna chain
region
hydrophilic
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CN108760730B (en
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徐金梦
张彦
黄煜真
李丽
杨红梅
崔康
于京华
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University of Jinan
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—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
    • G01N21/78—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 producing a change of colour
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis

Abstract

The invention discloses a kind of methods that paper substrate double mode detects magnesium ion.Hydrophobic region and hydrophilic region are prepared on paper using wax printing and laser cutting technique, and by screen printing technique, print three electrodes.Functionalization is carried out by the different zones to paper chip, utilize the difference in size between material particle size and paper fiber aperture, 3, the recognition reaction of the coloration and magnesium ion and its specific DNA enzyme of 3 '-diaminobenzidines, may be implemented that the visualization to magnesium ion is qualitative and electrochemical luminescence precisely detects.

Description

A kind of method of paper substrate double mode detection magnesium ion
Technical field
The present invention relates to a kind of methods that paper substrate double mode detects magnesium ion, belong to the detection technique field of magnesium ion.
Background technology
Magnesium is a kind of participation organism normal activities and the essential element of metabolic processes, calcium, phosphorus, iodine, Metabolism of the nutrients such as potassium, vitamin in human body also depends on magnesium to play.It can promote cell proliferation and Growth, maintains the integrality of tissue.Magnesium is to reduce the dominant catalyst of Blood Cholesterol, and maintain systaltic Important ion.Internal magnesium deficiency can lead to cardiac muscle fibre necrosis, cardiac output attenuating, arrhythmia etc..Magnesium exhaustion may further result in Insulin resistance and insulin secretion damage.Therefore, there is important meaning to health to the analysis of magnesium ion concentration and detection Justice.
However, the method for detection magnesium ion needs specific equipment at present, the measurement period is long, complicated for operation, many methods It can only rest in laboratory and cannot be used for putting into practice.Therefore, researching and developing easy, quick, sensitive, accurate detection method is It is highly important.Micro-fluidic paper chip has that manufacturing process is simple, portable, at low cost, biocompatibility as novel device The advantages that good with biodegradability, can be used for the fields such as disease detection, environmental quality monitoring and water analysis.Therefore, it grinds Study carefully and develop paper device important in inhibiting novel, practical, cheap, easy to operate.
Invention content
For presently, there are the above problem, technical problem to be solved by the invention is to provide a kind of double-mode paper-baseds to set Standby construction method realizes that and electrochemical luminescence qualitative to the visualization of magnesium ion is fixed by colorimetric method and Electrochemiluminescince Amount detection, it is characterized in that including the following steps：
1. utilizing Adobe Illustrator CS4 Software for Design hydrophobic wax print patterns on computers and utilizing solid state wax It is then melted on the A4 filter paper of its bulk print to cutting in heater plate to wax and permeates entire paper by printer Thickness forms hydrophobic wall, and pattern is as shown in Fig. 1, and wherein A is colorimetric area, and B is electrochemical luminescence detection zone；
2. using the method for silk-screen printing, by Ag/AgCl reference electrodes, carbon working electrode and carbon are printed onto paper respectively to electrode The device back side circle hydrophilic region a, b, c, and the area that prints electrode is less than reserved hydrophilic region area, pattern is as shown in Fig. 2； The white border circular areas that colorimetric area is surrounded by grey hydrophobic pattern penetrates into electrochemical luminescence for auxiliary liquid and detects region B, the white rectangle region surrounded by grey hydrophobic pattern are used for recording color element, and electrochemical luminescence detection zone is dredged by grey The white rectangle region that water patterns surround is used for three electrode of auxiliary communication；
3. preparing dendritic gold nanoparticle：80 mL secondary waters are heated to 90 DEG C, and it is 1% that 0.8 mL mass fractions, which are added, Chlorauric acid solution continues to be heated to 96 DEG C of 1 min of holding, is eventually adding the sodium citrate that 2.8 mL mass fractions are 1%, and add Hot 5 min postcoolings are to room temperature；The 20 above-mentioned solution of μ L are taken to be added drop-wise to the round hydrophilic working region of electrochemical luminescence detection zone, from It is primary that aforesaid operations are repeated after so drying；Continue to be added dropwise 20 μ L by 0.018 g hydroxylamine hydrochlorides, 1.0 mL secondary waters and 667 μ L The mixed solution that the chlorauric acid solution that mass fraction is 1% forms, repeatedly aforesaid operations are primary after natural drying；
4. preparing graphene oxide-Ag-N- (4- ammonia butyl)-N- ethyl different luminol nano-complexes：By 1.0 mL 0.02 Mol/L N- (4- ammonia butyl)-N- ethyl different luminols are rapidly joined by 2.0 mL, 10 mmol/L silver nitrate solutions, 500 μ L 2.0 mg/mL graphene oxide solutions, 5.0 mL secondary waters, 9.0 mL ethyl alcohol composition mixed solution and in room temperature magnetic agitation Lower reaction 12 hours, graphene oxide-Ag-N- (4- ammonia fourths are obtained by obtained solution ethyl alcohol and secondary water washing for several times Base)-N- ethyl different luminol nano-complexes；
5. preparing cadmiumsulfide quantum dot：172 μ L mercaptopropionic acids are added in the cadmium chloride solution of 40 mL, 20 mmol/L and are used in combination It is 10 that the sodium hydroxide solution of 1 mol/L, which adjusts its pH, continuously adds the thioacetamide of 20 mL, 20 mmol/L, and continue It dialyses at room temperature 24 hours in 80 DEG C of 10 h of reflux, obtained cadmium sulfide colloid after stirring 30 min and obtains cadmium sulfide quantum Point；
6. preparing Au-horseradish peroxidase-DNA chain 2- HKUST-1@Pt compounds
(1) HKUST-1@Pt nano-particles are prepared：It weighs 0.545 g Gerhardites to be dissolved in 7.5 mL secondary waters, claim It takes 0.264 g trimesic acids to be dissolved in 7.5 mL ethyl alcohol, makes it completely dissolved；Two kinds of solution obtained above is mixed simultaneously 30 min of magnetic agitation obtains homogeneous solution at room temperature, it is reacted 24 h in 120 DEG C, is cooled to room temperature, with ethyl alcohol and two Secondary water washing for several times, and reacts 10 hours under 80 DEG C of vacuum states and obtains metal-organic framework material HKUST-1, HKUST-1 For Cu3(BTC)2, BTC represents trimesic acid；The chloroplatinic acid that 1 mL mass fractions are 1% is taken to be added 1 mL 1mg/mL's In HKUST-1 and 20 min are ultrasonically treated, take 2 mL, 0.1 mol/L sodium borohydride solutions that above-mentioned mixed solution is added dropwise simultaneously 30 min are vigorously stirred, finally, obtained solution is centrifuged into 10 min with 12000 rpm and is obtained three times with secondary water washing HKUST-1@Pt nano-particles；
(2) Au nanocube-horseradish peroxidase-DNA chain 2-HKUST-1@Pt compounds are prepared：By 0.15 mL 0.1 Mol/L sodium borohydride solutions preserved in 4 DEG C rapidly join after 3 h by 6.25 mL, 1.0 mmol/L chlorauric acid solutions and In the mixed solution of 18.75 mL, 0.1 mol/L cetyl trimethylammonium bromide solutions composition, brown solution is obtained, it will It reacts 4 h under room temperature magnetic agitation, obtains Au seed solutions；By 5.0 μ L Au seed solutions, 50 μ L, 0.01 mol/L Copper-bath, the freshly prepd ascorbic acid solutions of 3.0 mL, 0.1 mol/L are added sequentially to by 5.0 mL, 2.0 mmol/L In the mixed solution of gold chloride and 20 mL, 0.02 mol/L cetyl trimethylammonium bromides composition, solution becomes after 5 min For aubergine, obtained solution is centrifuged into 10 min with 12000 rpm and is washed twice with secondary water, Au nano cubics are obtained Body；Take the buffer solution of 40 μ L 5 μm of ol/L DNA chain 2 and 1.0 mL pH 5.2,1.5 μ L, 10 mmol/L tri-（2- carboxylics Ethyl）Phosphine mixes and keeps 1 h；Above-mentioned mixed solution is mixed to simultaneously magnetic agitation 1 with 500 μ L Au nano cubic liquid solutions h；By 1 mg mL of obtained solution and 80 μ L-1Horseradish peroxidase mixes and 1.5 h of magnetic agitation obtains Au at room temperature Nanocube -2 compound of horseradish peroxidase-DNA chain；By above-mentioned compound and 1.0 mL HKUST-1@Pt nanoparticles Son mixing simultaneously keeps 4 h after 16 h of magnetic agitation in 4 DEG C at room temperature, finally obtains Au for several times with ethyl alcohol and secondary water washing Nanocube-horseradish peroxidase-DNA chain 2-HKUST-1@Pt nano-particle compounds；
7. 20 μ L graphene oxides-Ag-N- (4- ammonia butyl)-N- ethyl different luminol nano-complexes is taken to be added drop-wise to gold nano The region b and naturally dry of particle modification；20 μ L cadmiumsulfide quantum dots are added dropwise successively, the chitosan of 15 μ L, 0.05 wt % is solid Determine liquid and naturally dry；4 DEG C of 24 h of preservation after continuing dropwise addition 20 μ L DNA chain, 3 solution and being kept for 1 hour in 37 DEG C；Finally Extra DNA chain 3, the 3 base sequence of DNA chain are removed with the buffer solution flushing work region of 10 mmol/L pH 8.0 It arranges as shown in nucleotides sequence list, wherein its 3 ' terminal modified upper sulfydryl and 6 methylene；
8. taking 20 μ L by the buffer solution of 60 μ L, 1 μm of ol/L DNA chain, 1,500 μ L pH 8.0,500 μ L, 1 mM tri- （2- carboxyethyls）The mixed solution and 20 μ L Au nanocube-horseradish peroxidases-DNA chain 2-HKUST-1 of phosphine composition Compound is mixed and heated to 95 °C to form Mg2+ Specific DNA enzyme；Paper device colorimetric area is cut along dotted portion It cuts, is folded along bold portion, the 15 above-mentioned solution of μ L is taken to be added drop-wise to colorimetric area circle hydrophilic region and keep at room temperature 1.5 h；Continue the solution to be measured that 20 μ L of dropwise addition contain magnesium ion and keep 0.5 h in 37 °C and is special with it to complete magnesium ion Property DNA enzymatic between specific recognition be connected with Au- horseradish peroxidating since material particle size and paper fiber pore size are different 2 sequence of DNA chain of object enzyme can permeate paper chip and realize and 3 base pair complementarity of DNA chain, 1 base sequence of the DNA chain such as core Shown in nucleotide sequence table, wherein its 5 ' terminal modified upper amino and 6 methylene, 2 base sequence of DNA chain such as nucleotides sequence list institute Show, wherein its 5 ' terminal modified upper sulfydryl, 3 ' terminal modified upper amino and 6 methylene, and the 23rd base A represents gland from left to right Purine ribonucleic acid；
9. 20 μ L, 20 mmol/L, 3,3 '-diaminobenzidines is taken to be added dropwise in the hydrophilic colour developing item of region A rectangles, it is connected with 2 sequence of DNA chain of HKUST-1@Pt NPs cannot penetrate paper chip since grain size is more than paper chip aperture, continue to justify to region A 20 μ L, 5 mmol/L H are added dropwise in the hydrophilic working region of shape2O2, the buffer solution of 40 μ L pH 8.0, liquid is added dropwise after reacting 5 min The hydrophilic colour developing item of body rectangle measures colour developing length after 10 min；
10. rectangular area below paper device electrochemical luminescence detection zone is cut along dotted portion, carried out along bold portion Folding makes rectangle hydrophilic region be aligned with round hydrophilic region, by 20 μ L, 10 mmol/L hydrogen peroxide and 40 μ L pH 8.0 Buffer solution mixing is added drop-wise to rectangle hydrophilic region, and connection electrochemical workstation detects magnesium ion concentration；
11. drawing the standard curve of electrochemical luminescence intensity and develop the color length and magnesium ion concentration respectively, the survey of magnesium ion is completed It is fixed.
12. the overall size of above-mentioned paper device is the 40 mm-110 mm of mm-50 mm × 100, wherein colorimetric area is by hydrophobic wax The a diameter of 5-7 mm of round hydrophilic region of encirclement, longitudinal rectangle hydrophilic region width are 1-3 mm, are highly 44-46 mm, It is divided into 1 mm between rectangle hydrophilic region right side scale；The a diameter of 5.5-7.5 of electrochemical luminescence detection zone circle hydrophilic region a, b, c Mm, around rectangle hydrophobic region width be 29-31 mm, be highly 14-16 mm, rectangle hydrophilic region width is below 23-25 mm are highly 4.5-6.5 mm, around hydrophobic wax width be 2-5 mm.
Beneficial effects of the present invention：
1. visualization to magnesium ion can be achieved at the same time in a kind of paper substrate double mode sensor and electrochemical luminescence precisely detects, Magnesium ion specific DNA enzyme is made full use of, experimental cost is reduced.
2. the present invention realizes HKUST-1@Pt nanoparticles using paper fiber aperture and the difference in size of material particle size The separation of son and Au nanocubes-horseradish peroxidase, and it is respectively used to the visualization qualitative detection and electrification of magnesium ion Learn the quantitative detection that shines.
3. by change ion specific DNA enzyme, it can be achieved that analysis and detection to other heavy metal ion.
Description of the drawings：
1. attached drawing 1 is hydrophobic wax print pattern.Wherein, A is colorimetric area, and B is electrochemical luminescence detection zone.
2. attached drawing 2 is 3 electrodes in silk-screen printing on hydrophobic wax print pattern, region a, b, c print Ag/AgCl respectively Reference electrode, carbon working electrode and carbon are to electrode.
Specific implementation mode
The present invention is described in further detail in the following with reference to the drawings and specific embodiments.
Embodiment 1（Magnesium ion in tap water）
A kind of method easy to operate, detection speed is fast, low-cost paper substrate double mode detects magnesium ion, including following step Suddenly.
1. Adobe Illustrator CS4 Software for Design hydrophobic wax print patterns are utilized on computers, such as attached drawing 1 It is shown.
2. filter paper is cut into A4 sizes.
3. the hydrophobic wax printed drawings designed in printing step 1 on the filter paper cut out in step 2 using wax spray printer Case.
4. the A4 filter paper with hydrophobic wax pattern obtained in step 3 is heated 2 at 100 DEG C on hot plate Min makes wax melt and permeates the thickness of entire paper, forms hydrophobic wall.
5. paper device is cut along dotted portion, folded along bold portion.
6. growing dendritic gold nanoparticle in paper fiber using the method that document has been reported.
7. preparing graphene oxide-Ag-N- (4- ammonia butyl)-N- ethyl different luminols using the method that document has been reported Nano-complex.
8. preparing cadmiumsulfide quantum dot using the method that document has been reported.
9. preparing Au- horseradish peroxidases -2 compound of DNA chain and HKUST-1@Pt using the method that document has been reported Nano-particle.
10. configuring mixed solution.Mixed solution by 60 μ L, 1 μm of ol/L DNA chain, 1,500 μ L pH 8.0 buffering Solution, 500 μ L, 1 mM tri-（2- carboxyethyls）Phosphine forms.
11. Au -2 compound of horseradish peroxidase-DNA chain that step 9 obtains is received with 1.0 mL HKUST-1@Pt Rice corpuscles mixes and keeps 4 h in 4 DEG C after 16 h of magnetic agitation at room temperature, finally ethyl alcohol and secondary water washing is used to obtain for several times To Au nanocube-horseradish peroxidase-DNA chain 2- HKUST-1@Pt nano-particle compounds.
12. the mixed solution obtained in 20 μ L steps 10 is taken to be mixed and heated with the solution obtained in 20 μ L steps 11 To 95 °C to form Mg2+ Specific DNA enzyme.
13. taking graphene oxide-Ag-N- (4- ammonia the butyl)-N- ethyl different luminol nanometers obtained in 20 μ L steps 7 Compound is added drop-wise to the working region of gold nanoparticle modification and naturally dry, and 20 μ L cadmiumsulfide quantum dots are added dropwise in continuation successively, The chitosan fixer of 15 μ L, 0.05 wt % is in the round hydrophilic working region of electrochemical luminescence detection zone and naturally dry；After Continue and is preserved overnight for 4 DEG C after 20 μ L DNA chain, 3 solution is added dropwise and is kept for 1 hour in 37 DEG C；Finally with 10 mmol/L pH 8.0 Buffer solution flushing work region to remove extra DNA chain 3.
14. taking the solution obtained in 15 μ L steps 12 to be added drop-wise to colorimetric area circle hydrophilic region and keeping at room temperature 1.5 h continue the solution to be measured that 20 μ L of dropwise addition contain magnesium ion and keep 0.5 h in 37 °C, since material particle size and paper are fine Pore size difference is tieed up, paper chip realization and 3 base pair complementarity of DNA chain can be permeated by being connected with 2 sequence of DNA chain of Au-HRP.
15. being connected with 2 sequence of DNA chain of HKUST-1@Pt nano-particles since grain size cannot be saturating more than paper fiber aperture Paper chip is crossed, continues that 20 μ L, 5 mmol/L H are added dropwise to the round hydrophilic working region in colorimetric area2O2, 40 are added dropwise after reacting 5 min The buffer solution of μ L pH 8.0, liquid, which flows into be added dropwise, has the rectangle of 20 μ L, 20 mmol/L, 3,3 '-diaminobenzidines hydrophilic Develop the color item, and color element is measured after 10 min.
16. rectangular area below paper device electrochemical luminescence detection zone is cut along dotted portion, along bold portion Fold makes rectangle hydrophilic region be aligned with round hydrophilic region, by 20 μ L, 10 mmol/L hydrogen peroxide and 40 μ L pH 8.0 buffer solution mixing is added drop-wise to rectangle hydrophilic region, and connection electrochemical luminescence work station detects magnesium ion concentration.
17. take 20 μ L, 20 mmol/L 3,3',5,5'-tetramethylbenzidine in length be 2 mm, width be 15 mm Colorimetric strip area, and naturally dry.
18. drawing the standard curve of electrochemical luminescence intensity and develop the color length and magnesium ion concentration respectively, magnesium ion is completed Measurement.
19. the overall size of above-mentioned paper device is the 40 mm-110 mm of mm-50 mm × 100, wherein colorimetric area is by hydrophobic wax The a diameter of 5-7 mm of round hydrophilic region of encirclement, longitudinal rectangle hydrophilic region width are 1-3 mm, are highly 44-46 mm, It is divided into 1 mm between rectangle hydrophilic region right side scale；The a diameter of 5.5-7.5 of electrochemical luminescence detection zone circle hydrophilic region a, b, c Mm, around rectangle hydrophobic region width be 29-31 mm, be highly 14-16 mm, rectangle hydrophilic region width is below 23-25 mm are highly 4.5-6.5 mm, around hydrophobic wax width be 2-5 mm.
Sequence table
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gaagaagaag atac 14

Claims (4)

1. a kind of method of paper substrate double mode detection magnesium ion, it is characterized in that including the following steps：
(1) Adobe Illustrator CS4 Software for Design hydrophobic wax print patterns are utilized on computers and utilize solid state wax It is then melted on the A4 size filter paper of its bulk print to cutting to wax and is permeated entire in heater plate by printer The thickness of paper forms hydrophobic wall, and paper device is constructed by paper folding technology；The paper device includes two parts, and respectively electrochemistry is sent out Light detection area and colorimetric area, wherein electrochemical luminescence detection zone are printed on round hydrophilic region and rectangle hydrophilic region, are respectively intended to Three electrode of three electrodes and auxiliary communication is printed, colorimetric area is printed on round hydrophilic working region and rectangle is hydrophilic than vitta, uses respectively Carry out auxiliary liquid and penetrates into electrochemical luminescence detection zone circle hydrophilic region and record color element；
(2) method for using silk-screen printing, by carbon to electrode, carbon working electrode and Ag/AgCl reference electrodes are from left to right successively It is printed onto paper device back side electrochemical luminescence detection zone circle hydrophilic region, and the electrode area printed is less than reserved hydrophilic region Area, paper device can be cut along dotted portion, be folded along bold portion, and colorimetric area circle hydrophilic region and electricity can be made Round hydrophilic region overlaps among chemiluminescence detection area；
(3) functionalization is carried out to the round hydrophilic working region of electrochemical luminescence detection zone, is given birth to first by seed solution growth method Long dendritic gold nanoparticle, specific growth step are：80 mL secondary waters are heated to 90 DEG C, and 0.8 mL mass point is added Number is 1% chlorauric acid solution, continues to be heated to 96 DEG C of 1 min of holding, is eventually adding the lemon that 2.8 mL mass fractions are 1% Lemon acid sodium, and 5 min postcoolings are heated to room temperature；The 20 above-mentioned solution of μ L are taken to be added drop-wise to electrochemical luminescence detection zone circle hydrophilic Working region, repeatedly aforesaid operations are primary after natural drying；Continue to be added dropwise 20 μ L by 0.018 g hydroxylamine hydrochlorides, 1.0 mL bis- times The mixed solution that the chlorauric acid solution that water and 667 μ L mass fractions are 1% forms, repeatedly aforesaid operations are primary after natural drying；
(4) graphene oxide-Ag-N- (4- ammonia butyl)-N- second is modified in the round hydrophilic working region of electrochemical luminescence detection zone Base different luminol nano-complex, the specific steps are：By 1.0 mL, 0.02 mol/L N- (4- ammonia butyl) the different Rumi of-N- ethyls Promise is rapidly joined by 2.0 mL, 10 mmol/L silver nitrate solutions, 500 μ L, 2.0 mg/mL graphene oxide solutions, 5.0 mL Secondary water, the mixed solution of 9.0 mL ethyl alcohol composition simultaneously react 12 hours, the solution second that will be obtained under room temperature magnetic agitation Alcohol and secondary water washing obtain graphene oxide-Ag-N- (4- ammonia butyl)-N- ethyl different luminol nano-complexes for several times, take 20 μ L are added drop-wise to the working region of gold nanoparticle modification and naturally dry；
(5) continue to modify cadmiumsulfide quantum dot and DNA chain 3 in the round hydrophilic working region of electrochemical luminescence detection zone, specifically Step is：First, 172 μ L mercaptopropionic acids are added in the cadmium chloride solution of 40 mL 20 mmol/L and with the hydrogen of 1 mol/L It is 10 that sodium hydroxide solution, which adjusts its pH, continuously adds the thioacetamide of 20 mL, 20 mmol/L, and is continued after stirring 30 min Flow back 10 h in 80 DEG C, and obtained CdS colloids 24 h that dialyse at room temperature obtain cadmiumsulfide quantum dot；20 μ L sulphur are added dropwise successively Cadmium quantum dot, the chitosan fixer that 15 μ L mass fractions are 0.05 % is in the round hydrophilic work of electrochemical luminescence detection zone Region and naturally dry；4 DEG C of 24 h of preservation after continuing dropwise addition 20 μ L DNA chain, 3 solution and being kept for 1 hour in 37 DEG C；Finally Extra DNA chain 3, the 3 base sequence of DNA chain are removed with the buffer solution flushing work region of 10 mmol/L pH 8.0 It arranges as shown in nucleotides sequence list, wherein its 3 ' terminal modified upper sulfydryl and 6 methylene；
(6) in colorimetric area circle hydrophilic region modifying DNA chain 1 and Au nanocube-horseradish peroxidases-DNA chain 2- HKUST-1@Pt nano-particle compounds, the specific steps are：It weighs 0.545 g Gerhardites and is dissolved in 7.5 mL secondary waters In, it weighs 0.264 g trimesic acids and is dissolved in 7.5 mL ethyl alcohol, make it completely dissolved, by two kinds of solution obtained above It mixes and 30 min of magnetic agitation obtains homogeneous solution at room temperature, it is reacted into 24 h in 120 DEG C, is cooled to room temperature, uses second Alcohol and secondary water washing for several times, and keep obtaining metal-organic framework material HKUST-1 in 10 hours under 80 DEG C of vacuum states, HKUST-1 is Cu3(BTC)2, BTC represents trimesic acid；Take the chloroplatinic acid that 1 mL mass fractions are 1% that 1 mL, 1 mg/ are added In the HKUST-1 of mL and 20 min are ultrasonically treated, take 2 mL, 0.1 mol/L sodium borohydride solutions that above-mentioned mixing is added dropwise molten Liquid is simultaneously vigorously stirred 30 min, finally, obtained solution is centrifuged 10 min with 12000 rpm and is obtained three times with secondary water washing To HKUST-1@Pt nano-particles；By 0.15 mL, 0.1 mol/L sodium borohydride solutions in 4 DEG C preserve 3 h after rapidly join by 6.25 mL, 1.0 mmol/L chlorauric acid solutions and 18.75 mL, 0.1 mol/L cetyl trimethylammonium bromide solutions composition Mixed solution in, obtain brown solution, it reacted into 4 h under room temperature magnetic agitation, obtains Au seed solutions；By 5.0 μ L Au seed solutions, 50 μ L, 0.01 mol/L copper-baths, the freshly prepd ascorbic acid solutions of 3.0 mL, 0.1 mol/L It is added sequentially to by 0.02 mol/L cetyl trimethylammonium bromides of 5.0 mL, 2.0 mmol/L chlorauric acid solutions and 20 mL In the mixed solution of composition, solution becomes aubergine after 5 min, and obtained solution is centrifuged 10 min simultaneously with 12000 rpm It is washed twice with secondary water, obtains Au nanocubes；Take that 40 μ L, 5 μm of ol/L DNA chain 2 and 1.0 mL pH's 5.2 is slow Rush solution, 1.5 μ L, 10 mmol/L tri-（2- carboxyethyls）Phosphine mixes and keeps 1 h；By above-mentioned mixed solution and 500 μ L Au The mixing of nano cubic liquid solution and 1 h of magnetic agitation；By 1 mg mL of obtained solution and 80 μ L-1Horseradish peroxidase is mixed Merge 1.5 h of magnetic agitation at room temperature and obtains Au nanocubes -2 compound of horseradish peroxidase-DNA chain；It will be above-mentioned Compound mixes with 1.0 mL HKUST-1@Pt nano-particles and keeps 4 h in 4 DEG C after 16 h of magnetic agitation at room temperature, most Au nanocube-horseradish peroxidase-DNA chain 2- HKUST-1 Pt are obtained for several times with ethyl alcohol and secondary water washing afterwards to receive Rice corpuscles compound；Take 20 μ L by the buffer solution of 60 μ L, 1 μm of ol/L DNA chain, 1,500 μ L pH 8.0,500 μ L 1 MM tri-（2- carboxyethyls）The mixed solution and 20 μ L Au nanocube-horseradish peroxidases-DNA chain 2- of phosphine composition HKUST-1@Pt nano-particle compounds are mixed and heated to 95 °C to form Mg2+ Specific DNA enzyme；By paper device colorimetric area It is cut along dotted portion, is folded along bold portion, the 15 above-mentioned solution of μ L is taken to be added drop-wise to colorimetric area circle hydrophilic region And 1.5 h are kept at room temperature, continue the solution to be measured that 20 μ L of dropwise addition contain magnesium ion and keep 0.5 h to complete in 37 °C Specific recognition between magnesium ion and its specific DNA enzyme is connected with since material particle size is different with paper fiber pore size 2 sequence of DNA chain of Au- horseradish peroxidases can permeate paper chip and realize and 3 base pair complementarity of DNA chain, the DNA chain 1 base sequence is as shown in nucleotides sequence list, wherein its 5 ' terminal modified upper amino and 6 methylene, and 2 base sequence of DNA chain is such as Shown in nucleotides sequence list, wherein its 5 ' terminal modified upper sulfydryl, 3 ' terminal modified upper amino and 6 methylene, and from left to right 23 base A represent adenine ribonucleic acid；
(7) it takes 20 μ L, 20 mmol/L, 3,3 '-diaminobenzidines to be added dropwise in the hydrophilic colour developing item of colorimetric area rectangle, is connected with 2 sequence of DNA chain of HKUST-1@Pt nano-particles cannot penetrate paper chip since grain size is more than paper fiber aperture, continue to colorimetric 20 μ L, 5 mmol/L H are added dropwise in the round hydrophilic working region in area2O2, react 5 min after be added dropwise 40 μ L pH 8.0 buffering it is molten Liquid, liquid flow into colour developing item, colour developing length are measured after 10 min；
(8) rectangular area below paper device electrochemical luminescence detection zone is cut along dotted portion, is carried out along bold portion Folding makes rectangle hydrophilic region be aligned with round hydrophilic region, by 20 μ L, 10 mmol/L hydrogen peroxide and 40 μ L pH 8.0 Buffer solution mixing is added drop-wise to rectangle hydrophilic region, and connection electrochemical workstation detects magnesium ion concentration；
(9) standard curve for drawing electrochemical luminescence intensity and develop the color length and magnesium ion concentration respectively, completes the survey of magnesium ion It is fixed.
2. a kind of method of paper substrate double mode detection magnesium ion according to claim 1, it is characterized in that Pt nanometers of HKUST-1@ Particle composite material grain size is more than paper fiber aperture, and paper fiber aperture is 11 μm, and HKUST-1@Pt can be realized by this difference The separation of nano-particle and Au nanocubes-horseradish peroxidase.
3. a kind of method of paper substrate double mode detection magnesium ion according to claim 1, it is characterized in that the HKUST-1@of synthesis Pt nano compositions can catalyzing hydrogen peroxide reduction, to make 3,3 '-diaminobenzidines show brownish red.
4. a kind of method of paper substrate double mode detection magnesium ion according to claim 1, it is characterized in that the paper device is total Size is the 40 mm-110 mm of mm-50 mm × 100, a diameter of 5- of round hydrophilic region that wherein colorimetric area is surrounded by hydrophobic wax 7 mm, longitudinal rectangle hydrophilic region width are 1-3 mm, are highly 44-46 mm, 1 is divided between rectangle hydrophilic region right side scale mm；The a diameter of 5.5-7.5 mm of electrochemical luminescence detection zone circle hydrophilic region, around rectangle hydrophobic region width be 29-31 Mm is highly 14-16 mm, below rectangle hydrophilic region width be 23-25 mm, be highly 4.5-6.5 mm, around dredge The width of water wax is 2-5 mm.
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