CN108760730B - Paper-based dual-mode magnesium ion detection method - Google Patents
Paper-based dual-mode magnesium ion detection method Download PDFInfo
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- CN108760730B CN108760730B CN201810453724.XA CN201810453724A CN108760730B CN 108760730 B CN108760730 B CN 108760730B CN 201810453724 A CN201810453724 A CN 201810453724A CN 108760730 B CN108760730 B CN 108760730B
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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
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- G—PHYSICS
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- 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
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Abstract
The invention discloses a paper-based dual-mode magnesium ion detection method. The hydrophobic area and the hydrophilic area are prepared on the paper by utilizing a wax printing and laser cutting technology, and the three electrodes are printed by utilizing a screen printing technology. By functionalizing different areas of the paper chip and utilizing the size difference between the particle size of the material and the aperture of the paper fiber, the color development effect of 3, 3' -diaminobenzidine and the recognition effect of magnesium ions and specific DNA enzymes thereof, the visual qualitative and electrochemical luminescence accurate detection of the magnesium ions can be realized.
Description
Technical Field
The invention relates to a paper-based dual-mode magnesium ion detection method, and belongs to the technical field of magnesium ion detection.
Background
Magnesium is an essential element participating in normal life activities and metabolism of organisms, and the metabolism of nutrients such as calcium, phosphorus, iodine, potassium, vitamins and the like in human bodies is also performed by magnesium. It can promote cell proliferation and growth and maintain the integrity of human tissue. Magnesium is a major catalyst for lowering cholesterol in the blood and is also an important ion for maintaining cardiac contraction. Lack of magnesium in the body can lead to myocardial fiber necrosis, cardiac output reduction, heart rhythm disorder and the like. Magnesium depletion can also lead to impaired insulin resistance and insulin secretion. Therefore, the analysis and detection of the magnesium ion concentration are of great significance to human health.
However, the existing method for detecting magnesium ions needs special equipment, the measuring period is long, the operation is complex, and many methods only stay in a laboratory and cannot be used for practice. Therefore, it is very important to research and develop a simple, rapid, sensitive and accurate detection method. The microfluidic paper chip serving as novel equipment has the advantages of simple manufacturing process, easiness in carrying, low cost, good biocompatibility and biodegradability and the like, and can be used in the fields of disease detection, environmental quality monitoring, water quality analysis and the like. Therefore, it is important to research and develop a new, practical, inexpensive, and easy-to-operate paper device.
Disclosure of Invention
Aiming at the problems, the invention aims to provide a construction method of a dual-mode paper-based device, which realizes visual qualitative detection and electrochemical luminescence quantitative detection of magnesium ions by a colorimetric method and an electrochemical luminescence method and is characterized by comprising the following steps:
1. designing a hydrophobic wax printing pattern on a computer by using Adobe Illustrator CS4 software, printing the hydrophobic wax printing pattern on a cut A4 filter paper in batch by using a solid wax printer, and then heating the hydrophobic wax printing pattern on a heating plate until the wax is melted and permeates the whole thickness of the paper to form a hydrophobic wall, wherein the pattern is shown in figure 1, A is a colorimetric area, and B is an electrochemiluminescence detection area;
2. respectively printing an Ag/AgCl reference electrode, a carbon working electrode and a carbon counter electrode on circular hydrophilic areas a, b and c on the back surface of the paper device by adopting a screen printing method, wherein the area of the printing electrode is smaller than that of the reserved hydrophilic areas, and the pattern is shown in figure 2; the colorimetric area is a white circular area surrounded by a gray hydrophobic pattern and used for assisting liquid to permeate into the electrochemiluminescence detection area b, a white rectangular area surrounded by the gray hydrophobic pattern is used for recording the color length, and the electrochemiluminescence detection area is a white rectangular area surrounded by the gray hydrophobic pattern and used for assisting in communicating three electrodes;
3. preparing dendritic gold nanoparticles: heating 80 mL of secondary water to 90 ℃, adding 0.8 mL of chloroauric acid solution with the mass fraction of 1%, continuously heating to 96 ℃, keeping for 1 min, finally adding 2.8 mL of sodium citrate with the mass fraction of 1%, heating for 5 min, and cooling to room temperature; dripping 20 mu L of the solution into a circular hydrophilic working area of an electrochemiluminescence detection area, naturally airing, and repeating the operation once; continuously dropwise adding 20 mu L of mixed solution consisting of 0.018 g of hydroxylamine hydrochloride, 1.0 mL of secondary water and 667 mu L of chloroauric acid solution with the mass fraction of 1%, naturally airing, and repeating the operation once;
4. preparing a graphene oxide-Ag-N- (4-aminobutyl) -N-ethyl isoluminol nano compound: 1.0 mL of 0.02 mol/L N- (4-aminobutyl) -N-ethyl isoluminol is quickly added into a mixed solution consisting of 2.0 mL of 10 mmol/L silver nitrate solution, 500 mu L of 2.0 mg/mL graphene oxide solution, 5.0 mL of secondary water and 9.0 mL of ethanol, the mixed solution is reacted for 12 hours under room temperature magnetic stirring, and the obtained solution is washed by ethanol and the secondary water for a plurality of times to obtain a graphene oxide-Ag-N- (4-aminobutyl) -N-ethyl isoluminol nano compound;
5. preparing cadmium sulfide quantum dots: adding 172 mu L of mercaptopropionic acid into 40 mL of 20 mmol/L cadmium chloride solution, adjusting the pH value of the solution to 10 by using 1 mol/L sodium hydroxide solution, continuously adding 20 mL of 20 mmol/L thioacetamide, continuously stirring for 30 min, refluxing at 80 ℃ for 10 h, and dialyzing the obtained cadmium sulfide colloid at room temperature for 24 h to obtain cadmium sulfide quantum dots;
6. preparation of Au-horseradish peroxidase-DNA chain 2-HKUST-1@ Pt Complex
(1) Preparation of HKUST-1@ Pt nanoparticles: 0.545 g of copper nitrate trihydrate is weighed and dissolved in 7.5 mL of secondary water, and 0.264 g of trimesic acid is weighed and dissolved in 7.5 mL of ethanol to ensure that the trimesic acid is completely dissolved; mixing the two solutions, magnetically stirring at room temperature for 30 min to obtain uniform solution, reacting at 120 deg.C for 24 hr, cooling to room temperature, washing with ethanol and water twice, and reacting at 80 deg.C under vacuum for 10 hr to obtain metal organic frame material HKUST-1 (HKUST-1 is Cu)3(BTC)2BTC stands for trimesic acid; adding 1 mL of chloroplatinic acid with mass fraction of 1% into 1 mL of HKUST-1 with mass fraction of 1mg/mL, performing ultrasonic treatment for 20 min, and hydroborating 2 mL of 0.1 mol/LDropwise adding a sodium solution into the mixed solution, vigorously stirring for 30 min, and finally centrifuging the obtained solution at 12000 rpm for 10 min and washing with secondary water for three times to obtain HKUST-1@ Pt nanoparticles;
(2) preparing Au nanocubes-horseradish peroxidase-DNA chain 2-HKUST-1@ Pt complex: storing 0.15 mL of 0.1 mol/L sodium borohydride solution at 4 ℃ for 3 h, quickly adding the sodium borohydride solution into a mixed solution consisting of 6.25 mL of 1.0 mmol/L chloroauric acid solution and 18.75 mL of 0.1 mol/L hexadecyl trimethyl ammonium bromide solution to obtain a yellow brown solution, and reacting the yellow brown solution for 4 h under magnetic stirring at room temperature to obtain an Au seed solution; adding 5.0 mu L of Au seed solution, 50 mu L of 0.01 mol/L copper sulfate solution and 3.0 mL of 0.1 mol/L newly prepared ascorbic acid solution into a mixed solution consisting of 5.0 mL of 2.0 mmol/L chloroauric acid and 20 mL of 0.02 mol/L hexadecyl trimethyl ammonium bromide in sequence, wherein the solution becomes purple red after 5 min, centrifuging the obtained solution at 12000 rpm for 10 min, and washing the solution twice with secondary water to obtain Au nanocubes; 40 μ L of 5 μmol/L DNA strand 2 was mixed with 1.0 mL of pH 5.2 buffer solution, 1.5 μ L of 10 mmol/L tris (2-carboxyethyl) phosphine and held for 1 h; mixing the mixed solution with 500 mu L of Au nanocube solution and magnetically stirring for 1 h; the resulting solution was mixed with 80. mu.L of 1mg mL-1Mixing horseradish peroxidase and magnetically stirring for 1.5 h at room temperature to obtain an Au nanocube-horseradish peroxidase-DNA chain 2 compound; mixing the complex with 1.0 mL of HKUST-1@ Pt nano particles, magnetically stirring at room temperature for 16 h, keeping at 4 ℃ for 4 h, and finally washing with ethanol and secondary water for several times to obtain an Au nanocube-horseradish peroxidase-DNA chain 2-HKUST-1@ Pt nano particle complex;
7. taking 20 mu L of graphene oxide-Ag-N- (4-aminobutyl) -N-ethyl isoluminol nano compound, dropwise adding the graphene oxide-Ag-N- (4-aminobutyl) -N-ethyl isoluminol nano compound to the gold nanoparticle modified area b, and naturally airing; sequentially dripping 20 mu L of cadmium sulfide quantum dots and 15 mu L of 0.05 wt% of chitosan fixing solution and naturally airing; continuously dropwise adding 20 mu L of DNA chain 3 solution, keeping the solution at 37 ℃ for 1 hour, and storing the solution at 4 ℃ for 24 hours; finally, washing the working area by using 10 mmol/L buffer solution with the pH value of 8.0 to remove redundant DNA chain 3, wherein the base sequence of the DNA chain 3 is shown in a nucleotide sequence table, and the 3' end of the DNA chain is modified with sulfydryl and 6 methylene;
8. mu.L of a mixed solution consisting of 60. mu.L of 1. mu. mol/L DNA strand 1, 500. mu.L of a buffer solution of pH 8.0, 500. mu.L of 1 mM tris (2-carboxyethyl) phosphine was mixed with 20. mu.L of Au nanocubes-horseradish peroxidase-DNA strand 2-HKUST-1 complex and heated to 95 ℃ to form Mg2+ A specific dnase; cutting the colorimetric region of the paper device along the dotted line part, folding along the solid line part, taking 15 mu L of the solution, dropwise adding the solution into the circular hydrophilic region of the colorimetric region, and keeping the solution at room temperature for 1.5 h; continuously dropwise adding 20 mu L of solution to be detected containing magnesium ions, and keeping the temperature at 37 ℃ for 0.5 h to finish the specific identification between the magnesium ions and the specific DNA enzyme thereof, wherein the particle size of the material is different from the pore size of paper fibers, a DNA chain 2 sequence connected with Au-horseradish peroxidase can permeate a paper chip to realize base complementary pairing with a DNA chain 3, the base sequence of the DNA chain 1 is shown in a nucleotide sequence table, the 5 ' end of the DNA chain is modified with amino and 6 methylene, the base sequence of the DNA chain 2 is shown in the nucleotide sequence table, the 5 ' end of the DNA chain is modified with mercapto, the 3 ' end of the DNA chain is modified with amino and 6 methylene, and the 23 rd base A from left to right represents adenine ribonucleic acid;
9. 20 mu.L of 20 mmol/L3, 3' -diaminobenzidine is dripped into the rectangular hydrophilic color development strip in the area A, the DNA chain 2 sequence connected with the HKUST-1@ Pt NPs cannot penetrate through the paper chip because the particle diameter is larger than the aperture of the paper chip, and 20 mu.L of 5 mmol/L H is dripped into the circular hydrophilic working area in the area A continuously2O2After reacting for 5 min, dripping 40 mu L of buffer solution with pH of 8.0 and liquid rectangular hydrophilic color development strips, and measuring the color development length after 10 min;
10. cutting a rectangular area below the electrochemiluminescence detection area of the paper device along the dotted line part, folding along the solid line part to enable the rectangular hydrophilic area to be aligned with the round hydrophilic area, mixing 20 mu L of 10 mmol/L hydrogen peroxide and 40 mu L of buffer solution with the pH value of 8.0, dropwise adding the mixture into the rectangular hydrophilic area, and connecting an electrochemical workstation to detect the concentration of magnesium ions;
11. and respectively drawing standard curves of the electrochemiluminescence intensity, the color development length and the magnesium ion concentration to finish the determination of the magnesium ions.
12. The total size of the paper device is 40 mm-50 mm multiplied by 100 mm-110 mm, wherein the diameter of a circular hydrophilic area surrounded by hydrophobic wax in the colorimetric area is 5-7 mm, the width of a longitudinal rectangular hydrophilic area is 1-3 mm, the height of the longitudinal rectangular hydrophilic area is 44-46 mm, and the scale interval on the right side of the rectangular hydrophilic area is 1 mm; the diameters of the circular hydrophilic areas a, b and c of the electrochemiluminescence detection area are 5.5-7.5 mm, the width of the rectangular hydrophobic area around the circular hydrophilic areas is 29-31 mm, the height of the rectangular hydrophilic area is 14-16 mm, the width of the rectangular hydrophilic area below the circular hydrophilic area is 23-25 mm, the height of the rectangular hydrophilic area is 4.5-6.5 mm, and the width of the hydrophobic wax around the rectangular hydrophilic area is 2-5 mm.
The invention has the beneficial effects that:
1. the paper-based dual-mode sensor can simultaneously realize the visualization and the accurate electrochemical luminescence detection of magnesium ions, fully utilizes the magnesium ion specific DNA enzyme and reduces the experiment cost.
2. The separation of the HKUST-1@ Pt nano particles and the Au nano cube-horseradish peroxidase is realized by utilizing the size difference of the paper fiber aperture and the material particle size, and the method is respectively used for visual qualitative detection and electrochemical luminescence quantitative detection of magnesium ions.
3. By changing the ion-specific DNA enzyme, the analysis and detection of other heavy metal ions can be realized.
Description of the drawings:
1. figure 1 is a hydrophobic wax print pattern. Wherein, A is a colorimetric zone and B is an electrochemiluminescence detection zone.
2. FIG. 2 shows that 3 electrodes are screen-printed on the hydrophobic wax printing pattern, and Ag/AgCl reference electrodes, carbon working electrodes and carbon counter electrodes are printed in the areas a, b and c, respectively.
Detailed Description
The invention is described in further detail below with reference to the figures and specific embodiments.
Example 1 (magnesium ion in tap water)
The paper-based dual-mode magnesium ion detection method is simple to operate, high in detection speed and low in cost and comprises the following steps.
1. The hydrophobic wax print pattern was designed on a computer using Adobe Illustrator CS4 software as shown in figure 1.
2. The filter paper was cut to a size of a 4.
3. And (3) printing the hydrophobic wax printing pattern designed in the step (1) on the filter paper cut in the step (2) by using a wax spraying printer.
4. And (3) heating the A4 filter paper with the hydrophobic wax pattern obtained in the step (3) on a heating plate at 100 ℃ for 2 min to melt the wax and penetrate the whole thickness of the paper to form the hydrophobic wall.
5. The paper device is cut along the dotted line portion and folded along the solid line portion.
6. Dendritic gold nanoparticles are grown on the paper fibers by a method reported in the literature.
7. The graphene oxide-Ag-N- (4-aminobutyl) -N-ethyl isoluminol nano-composite is prepared by a method reported in a literature.
8. The cadmium sulfide quantum dots are prepared by a method reported in the literature.
9. The Au-horseradish peroxidase-DNA chain 2 complex and the HKUST-1@ Pt nano-particle are prepared by the method reported in the literature.
10. And preparing a mixed solution. The mixed solution consisted of 60. mu.L of 1. mu. mol/L of DNA strand 1, 500. mu.L of a buffer solution of pH 8.0, 500. mu.L of 1 mM tris (2-carboxyethyl) phosphine.
11. And (3) mixing the Au-horseradish peroxidase-DNA chain 2 complex obtained in the step (9) with 1.0 mL of HKUST-1@ Pt nano particles, magnetically stirring at room temperature for 16 h, keeping the temperature at 4 ℃ for 4 h, and finally washing with ethanol and secondary water for several times to obtain the Au nanocube-horseradish peroxidase-DNA chain 2-HKUST-1@ Pt nano particle complex.
12. Mixing 20. mu.L of the mixed solution obtained in step 10 with 20. mu.L of the solution obtained in step 11 and heating to 95 ℃ to form Mg2+ Specific dnase.
13. Taking 20 mu L of the graphene oxide-Ag-N- (4-aminobutyl) -N-ethyl isoluminol nano compound obtained in the step 7, dropwise adding the graphene oxide-Ag-N- (4-aminobutyl) -N-ethyl isoluminol nano compound into a gold nanoparticle modified working area, naturally airing, continuously and sequentially dropwise adding 20 mu L of cadmium sulfide quantum dots and 15 mu L of 0.05 wt% of chitosan fixing solution into a circular hydrophilic working area of an electrochemiluminescence detection area, and naturally airing; continuously dropwise adding 20 mu L of DNA chain 3 solution, keeping the solution at 37 ℃ for 1 hour, and then storing the solution at 4 ℃ overnight; finally, the working area was washed with 10 mmol/L of a buffer solution of pH 8.0 to remove excess DNA strand 3.
14. And (3) dropwise adding 15 mu L of the solution obtained in the step (12) into a circular hydrophilic area of the colorimetric area, keeping the temperature for 1.5 h, continuously dropwise adding 20 mu L of the solution to be detected containing magnesium ions, keeping the temperature for 0.5 h at 37 ℃, and allowing the DNA chain 2 sequence connected with the Au-HRP to permeate the paper chip to realize base complementary pairing with the DNA chain 3 due to different particle sizes of the material and the paper fiber aperture.
15. The DNA chain 2 sequence connected with the HKUST-1@ Pt nano particle can not penetrate through the paper chip because the particle diameter is larger than the aperture of the paper fiber, and 20 mu L of 5 mmol/L H is continuously dripped into the circular hydrophilic working area of the colorimetric area2O2After 5 min of reaction, 40. mu.L of a buffer solution of pH 8.0 was added dropwise, and the liquid was poured into a rectangular hydrophilic color developing strip to which 20. mu.L of 20 mmol/L3, 3' -diaminobenzidine was added dropwise, and the color length was measured after 10 min.
16. Cutting a rectangular area below the electrochemiluminescence detection area of the paper device along the dotted line part, folding along the solid line part to enable the rectangular hydrophilic area to be aligned with the round hydrophilic area, mixing 20 mu L of 10 mmol/L hydrogen peroxide with 40 mu L of buffer solution with the pH value of 8.0, dropwise adding the mixture into the rectangular hydrophilic area, and connecting the electrochemiluminescence workstation to detect the concentration of magnesium ions.
17. 20 μ L of 20 mmol/L3, 3 ', 5, 5' -tetramethylbenzidine was placed in a 2 mm long and 15 mm wide colorimetric strip area and allowed to air dry.
18. And respectively drawing standard curves of the electrochemiluminescence intensity, the color development length and the magnesium ion concentration to finish the determination of the magnesium ions.
19. The total size of the paper device is 40 mm-50 mm multiplied by 100 mm-110 mm, wherein the diameter of a circular hydrophilic area surrounded by hydrophobic wax in the colorimetric area is 5-7 mm, the width of a longitudinal rectangular hydrophilic area is 1-3 mm, the height of the longitudinal rectangular hydrophilic area is 44-46 mm, and the scale interval on the right side of the rectangular hydrophilic area is 1 mm; the diameters of the circular hydrophilic areas a, b and c of the electrochemiluminescence detection area are 5.5-7.5 mm, the width of the rectangular hydrophobic area around the circular hydrophilic areas is 29-31 mm, the height of the rectangular hydrophilic area is 14-16 mm, the width of the rectangular hydrophilic area below the circular hydrophilic area is 23-25 mm, the height of the rectangular hydrophilic area is 4.5-6.5 mm, and the width of the hydrophobic wax around the rectangular hydrophilic area is 2-5 mm.
Sequence listing
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Claims (4)
1. The paper-based dual-mode magnesium ion detection method is characterized by comprising the following steps of:
(1) designing a hydrophobic wax printing pattern on a computer by using Adobe Illustrator CS4 software, printing the hydrophobic wax printing pattern on cut A4-sized filter paper in batches by using a solid wax printer, heating the filter paper on a heating plate until the wax is melted and penetrates through the thickness of the whole paper to form a hydrophobic wall, and constructing a paper device by using a paper folding technology; the paper device comprises two parts, namely an electrochemiluminescence detection area and a colorimetric area, wherein the electrochemiluminescence detection area is printed with a round hydrophilic area and a rectangular hydrophilic area which are respectively used for printing three electrodes and assisting in communicating the three electrodes, and the colorimetric area is printed with a round hydrophilic working area and a rectangular hydrophilic colorimetric strip which are respectively used for assisting liquid to permeate into the round hydrophilic area of the electrochemiluminescence detection area and recording the color length;
(2) printing a carbon counter electrode, a carbon working electrode and an Ag/AgCl reference electrode on a circular hydrophilic area of an electrochemical luminescence detection area on the back of a paper device from left to right in sequence by adopting a screen printing method, wherein the area of the printed electrode is smaller than that of a reserved hydrophilic area, the paper device is cut along a dotted line part and folded along a solid line part, so that the circular hydrophilic working area of a colorimetric area is superposed with the middle circular hydrophilic area of the electrochemical luminescence detection area;
(3) functionalizing a circular hydrophilic area of an electrochemiluminescence detection area, firstly growing dendritic gold nanoparticles by a seed solution growth method, wherein the specific growth steps are as follows: heating 80 mL of secondary water to 90 ℃, adding 0.8 mL of chloroauric acid solution with the mass fraction of 1%, continuously heating to 96 ℃, keeping for 1 min, finally adding 2.8 mL of sodium citrate with the mass fraction of 1%, heating for 5 min, and cooling to room temperature; dripping 20 mu L of the solution into a circular hydrophilic area of an electrochemiluminescence detection area, naturally airing, and repeating the operation once; continuously dropwise adding 20 mu L of mixed solution consisting of 0.018 g of hydroxylamine hydrochloride, 1.0 mL of secondary water and 667 mu L of chloroauric acid solution with the mass fraction of 1%, naturally airing, and repeating the operation once;
(4) modifying a graphene oxide-Ag-N- (4-aminobutyl) -N-ethyl isoluminol nano compound in a circular hydrophilic area of an electrochemiluminescence detection area, and specifically comprises the following steps: 1.0 mL of 0.02 mol/L N- (4-aminobutyl) -N-ethyl isoluminol is quickly added into a mixed solution consisting of 2.0 mL of 10 mmol/L silver nitrate solution, 500 mu L of 2.0 mg/mL graphene oxide solution, 5.0 mL of secondary water and 9.0 mL of ethanol, the mixed solution is magnetically stirred at room temperature for reaction for 12 hours, the obtained solution is washed by ethanol and the secondary water for a plurality of times to obtain a graphene oxide-Ag-N- (4-aminobutyl) -N-ethyl isoluminol nano compound, 20 mu L of the nano compound is taken and dripped into a round hydrophilic area of an electrochemical luminescence detection area modified by gold nanoparticles, and the nano compound is naturally aired;
(5) continuously modifying cadmium sulfide quantum dots and DNA chains 3 in a circular hydrophilic area of an electrochemiluminescence detection area, and specifically comprising the following steps: firstly, adding 172 mu L mercaptopropionic acid into 40 mL of 20 mmol/L cadmium chloride solution, adjusting the pH value of the solution to 10 by using 1 mol/L sodium hydroxide solution, continuously adding 20 mL of 20 mmol/L thioacetamide, continuously stirring for 30 min, refluxing for 10 h at 80 ℃, and dialyzing the obtained CdS colloid for 24 h at room temperature to obtain cadmium sulfide quantum dots; sequentially dripping 20 mu L of cadmium sulfide quantum dots and 15 mu L of chitosan fixing solution with the mass fraction of 0.05 percent into a circular hydrophilic area of an electrochemiluminescence detection area, and naturally airing; continuously dropwise adding 20 mu L of DNA chain 3 solution, keeping the solution at 37 ℃ for 1 hour, and storing the solution at 4 ℃ for 24 hours; finally, washing the circular hydrophilic area of the electrochemiluminescence detection area by using 10 mmol/L buffer solution with the pH value of 8.0 to remove redundant DNA chains 3, wherein the base sequence of the DNA chains 3 is shown in a nucleotide sequence table, and the 3' end of the DNA chains is modified with sulfydryl and 6 methylene groups;
(6) a DNA chain 1 and Au nanocubes-horseradish peroxidase-DNA chain 2-HKUST-1@ Pt nanoparticle compound is modified in a circular hydrophilic working area of a colorimetric area, and the specific steps are as follows: weighing 0.545 g of copper nitrate trihydrate to be dissolved in 7.5 mL of secondary water, weighing 0.264 g of trimesic acid to be dissolved in 7.5 mL of ethanol to be completely dissolved, mixing the two solutions obtained above, magnetically stirring for 30 min at room temperature to obtain a uniform solution, reacting at 120 ℃ for 24 h, cooling to room temperature, washing with ethanol and secondary water for a plurality of times, and keeping the solution at 80 ℃ for 10 hours in a vacuum state to obtain a metal organic framework material HKUST-1, wherein HKUST-1 is Cu3(BTC)2BTC stands for trimesic acid; adding 1 mL of chloroplatinic acid with the mass fraction of 1% into 1 mL of HKUST-1 with the mass fraction of 1mg/mL, carrying out ultrasonic treatment for 20 min, dropwise adding 2 mL of 0.1 mol/L sodium borohydride solution into the mixed solution, violently stirring for 30 min, and finally, centrifuging the obtained solution at 12000 rpm for 10 min and washing the solution with secondary water for three times to obtain HKUST-1@ Pt nano particles; 0.15 mL of 0.1 mol/L sodium borohydride solution was stored at 4 ℃ for 3 hours and then added rapidly with 6.25 mL of 1.0 mmol/L chloroauric acid solution and 18.75 mL of 0.1 mIn a mixed solution composed of ol/L hexadecyl trimethyl ammonium bromide solution, obtaining a yellow brown solution, and reacting the yellow brown solution for 4 hours under room temperature magnetic stirring to obtain Au seed solution; adding 5.0 mu L of Au seed solution, 50 mu L of 0.01 mol/L copper sulfate solution and 3.0 mL of 0.1 mol/L newly prepared ascorbic acid solution into a mixed solution consisting of 5.0 mL of 2.0 mmol/L chloroauric acid solution and 20 mL of 0.02 mol/L hexadecyl trimethyl ammonium bromide in sequence, wherein the solution becomes mauve after 5 min, centrifuging the obtained solution at 12000 rpm for 10 min, and washing the solution twice with secondary water to obtain Au nanocubes; 40 μ L of 5 μmol/L DNA strand 2 was mixed with 1.0 mL of pH 5.2 buffer solution, 1.5 μ L of 10 mmol/L tris (2-carboxyethyl) phosphine and held for 1 h; mixing the mixed solution with 500 mu L of Au nanocube solution and magnetically stirring for 1 h; the resulting solution was mixed with 80. mu.L of 1mg mL-1Mixing horseradish peroxidase and magnetically stirring for 1.5 h at room temperature to obtain an Au nanocube-horseradish peroxidase-DNA chain 2 compound; mixing the complex with 1.0 mL of HKUST-1@ Pt nano particles, magnetically stirring at room temperature for 16 h, keeping at 4 ℃ for 4 h, and finally washing with ethanol and secondary water for several times to obtain an Au nanocube-horseradish peroxidase-DNA chain 2-HKUST-1@ Pt nano particle complex; mu.L of a mixed solution consisting of 60. mu.L of 1. mu. mol/L DNA strand 1, 500. mu.L of a buffer solution of pH 8.0, 500. mu.L of 1 mM tris (2-carboxyethyl) phosphine was mixed with 20. mu.L of Au nanocubes-horseradish peroxidase-DNA strand 2-HKUST-1@ Pt nanoparticle complex and heated to 95 ℃ to form Mg2+ A specific dnase; cutting a color comparison area of a paper device along a dotted line part, folding along a solid line part, dropwise adding 15 mu L of the solution into a circular hydrophilic working area of the color comparison area, keeping the temperature at room temperature for 1.5 h, continuously dropwise adding 20 mu L of a solution to be detected containing magnesium ions, and keeping the temperature at 37 ℃ for 0.5 h to finish specific identification between the magnesium ions and specific DNA enzymes thereof, wherein due to the difference between the particle size of materials and the pore size of paper fibers, a DNA chain 2 sequence connected with Au-horseradish peroxidase can permeate a paper chip to realize base complementary pairing with a DNA chain 3, the base sequence of the DNA chain 1 is shown as a nucleotide sequence table, the 5' end of the DNA chain is modified with amino and 6 methylene, and the base sequence of the DNA chain 2 is shown as the nucleotide sequence table, wherein the base sequence of the DNA chainThe 5 'end of the compound is modified with sulfydryl, the 3' end of the compound is modified with amino and 6 methylene, and the 24 th base A from left to right represents adenine ribonucleic acid;
(7) 20 mu L of 20 mmol/L3, 3' -diaminobenzidine is dripped into a rectangular hydrophilic colorimetric strip in the colorimetric area, the DNA chain 2 sequence connected with the HKUST-1@ Pt nano particles cannot penetrate through the paper chip because the particle size is larger than the aperture of paper fiber, and 20 mu L of 5 mmol/L H is dripped into a circular hydrophilic working area in the colorimetric area continuously2O2After reacting for 5 min, dripping 40 mu L of buffer solution with pH of 8.0, enabling the liquid to flow into a rectangular hydrophilic colorimetric strip in the colorimetric area, and measuring the color development length after 10 min;
(8) cutting a rectangular area below the electrochemiluminescence detection area of the paper device along the dotted line part, folding along the solid line part to enable the rectangular hydrophilic area to be aligned with the round hydrophilic area, mixing 20 mu L of 10 mmol/L hydrogen peroxide and 40 mu L of buffer solution with the pH value of 8.0, dropwise adding the mixture into the rectangular hydrophilic area, and connecting an electrochemical workstation to detect the concentration of magnesium ions;
(9) and respectively drawing standard curves of the electrochemiluminescence intensity, the color development length and the magnesium ion concentration to finish the determination of the magnesium ions.
2. The paper-based dual-mode magnesium ion detection method according to claim 1, characterized in that the particle size of the HKUST-1@ Pt nanoparticle composite material is larger than the pore size of a paper fiber, the pore size of the paper fiber is 11 μm, and the separation of the HKUST-1@ Pt nanoparticles and Au nanocubes-horseradish peroxidase can be realized by means of the difference.
3. The paper-based dual-mode magnesium ion detection method according to claim 1, wherein the synthesized HKUST-1@ Pt nanoparticle composite material can catalyze hydrogen peroxide reduction, so that 3, 3' -diaminobenzidine is brownish red.
4. The paper-based dual-mode magnesium ion detection method according to claim 1, characterized in that the overall size of the paper device is 40 mm-50 mm x 100 mm-110 mm, wherein the diameter of a circular hydrophilic working area surrounded by hydrophobic wax in a colorimetric area is 5-7 mm, the width of a longitudinal rectangular hydrophilic colorimetric strip is 1-3 mm, the height is 44-46 mm, and the scale interval on the right side of the rectangular hydrophilic colorimetric strip is 1 mm; the diameter of the circular hydrophilic area of the electrochemiluminescence detection area is 5.5-7.5 mm, the width of the rectangular hydrophobic area around the circular hydrophilic area is 29-31 mm, the height of the rectangular hydrophobic area is 14-16 mm, the width of the rectangular hydrophilic area below the circular hydrophilic area is 23-25 mm, the height of the rectangular hydrophilic area is 4.5-6.5 mm, and the width of the hydrophobic wax around the rectangular hydrophilic area is 2-5 mm.
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