CN113514524B - Potential sensing array capable of calibrating reference potential and response slope simultaneously and preparation method thereof - Google Patents
Potential sensing array capable of calibrating reference potential and response slope simultaneously and preparation method thereof Download PDFInfo
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- CN113514524B CN113514524B CN202110355522.3A CN202110355522A CN113514524B CN 113514524 B CN113514524 B CN 113514524B CN 202110355522 A CN202110355522 A CN 202110355522A CN 113514524 B CN113514524 B CN 113514524B
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- 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
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3271—Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
- G01N27/3272—Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels
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- 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
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3271—Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
- G01N27/3274—Corrective measures, e.g. error detection, compensation for temperature or hematocrit, calibration
Abstract
The invention discloses a potential sensing array for simultaneously calibrating a reference potential and a response slope and application thereof. The invention designs a reference potential calibration channel in the potential sensing array to automatically calibrate the reference potential error caused by nonspecific adsorption in the actual sample test, thereby improving the test accuracy. The pH sensing channel is designed in the potential sensing array to calibrate the change of the target sensing channel to the potential response slope of the target object caused by pH, so that the pH sensor is suitable for testing body fluid with wide pH range. The potential sensing array has important significance in the aspect of development of intelligent toilets for home self-health monitoring.
Description
Technical Field
The invention relates to a sensor array, in particular to a potential sensing array for calibrating a reference potential and a response slope simultaneously and application thereof.
Background
The household sensing equipment can monitor the development condition of diseases for a long time and is a very effective disease monitoring auxiliary tool. The home sensing device can not only reduce the medical burden of families and society, but also provide the long-term health condition of patients in clinic. The medical service system is transformed to the family-based direction, so that the medical service system has great social application value. Compared with other sensing technologies, the potential sensing technology is more suitable for preparing household sensing equipment because the instrument is simple and portable, the price is low, and the miniaturization is easy.
However, the application of potential sensing technology in home smart devices still faces many challenges. First, home-use sensing devices require simple operation and portable instruments, and thus cannot pre-process samples. The home smart device requires that the reference potential of the sensor as a front-end sensing tool is not interfered by the potential caused by non-specific adsorption in the actual sample test. Secondly, the response slope of the sensor to the target is required to be constant or to be automatically calibrated in a wide physiological pH range, and only the sensing equipment meeting the two requirements can accurately convert the test signal into the concentration of the target. Therefore, how to simultaneously calibrate the reference potential and the response slope to improve the test accuracy without preprocessing the sample is a problem to be solved urgently.
Disclosure of Invention
In view of the above problems of the prior art, it is an object of the present invention to develop a potentiometric sensor array that simultaneously calibrates a reference potential and a response slope, and is used for detecting the concentration of a disease indicator in a body fluid.
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of a potential sensing array for calibrating a reference potential and a response slope simultaneously comprises the following specific steps:
step 1: preparing a basic electrode array:
firstly, designing a basic electrode array pattern on a substrate material, and preparing a basic electrode array by a printing process; the basic electrode array comprises a row of silver/silver chloride electrode channels and at least three rows of carbon-based electrode channels; each column of electrode channels comprises a circuit channel, a USB interface and an electrode sensing area; the printing process specifically comprises the following steps: printing carbon ink on a circuit channel, a USB interface and an electrode sensing area in each row of electrode channels, and drying in a drying oven at 50-200 ℃ for 3-10 h; then selecting an electrode channel printed with carbon ink, and printing the silver/silver chloride ink on a sensing area of the electrode channel to obtain a silver/silver chloride electrode; finally, printing the insulating paste on all substrate materials except the USB interface and the electrode sensing area, and drying in an oven at 50-200 ℃ for 3-10 h;
the substrate material is synthetic paper, polyethylene terephthalate film or textile;
the printing process is screen printing, ink-jet printing or manual coating;
step 2: modifying the basic electrode array to obtain a potential sensing array which simultaneously calibrates a reference potential and a response slope:
(1) preparing a polyvinyl butyral resin reference film solution: uniformly mixing sodium chloride, polyvinyl butyral resin and anhydrous methanol according to the mass-volume ratio of 50-100 mg: 100-250 mg: 2mL to obtain a polyvinyl butyral resin reference membrane solution; dripping polyvinyl butyral resin reference film solution on the surface of a silver/silver chloride electrode of a basic electrode array to form a reference electrode channel; wherein the thickness of the reference film of the polyvinyl butyral resin is 5-50 mu m;
(2) modifying a target sensing material on the surface of any column of carbon-based electrodes in the basic electrode array to form a target sensing electrode channel; modifying a reference potential calibration material on the surface of any column of carbon-based electrodes in the basic electrode array to form a reference potential calibration channel; modifying the pH sensing material on the surface of any column of carbon-based electrodes in the basic electrode array to form a pH sensing electrode channel; vacuum drying at 10-37 deg.C for 2-10h to obtain a potential sensing array with calibration reference potential and response slope, and storing at 0-10 deg.C;
the target sensing material is biological enzyme or nano enzyme;
the reference potential calibration material is a biological enzyme or nano-enzyme without catalytic activity;
the pH sensing material is polyaniline or a pH ion selective membrane;
the modification method is a dropping coating method or an electrochemical deposition method.
A potential sensing array which is manufactured by the method and simultaneously calibrates the reference potential and the response slope is provided.
The electric potential sensing array for simultaneously calibrating the reference electric potential and the response slope at least comprises a row of reference electrode channels, a row of pH sensing electrode channels, a row of target sensing electrode channels and a row of reference electric potential calibration channels.
The application of the potential sensing array for simultaneously calibrating the reference potential and the response slope in detecting the concentration of the disease indicator in the body fluid.
The invention designs a reference potential calibration channel in the potential sensing array to automatically calibrate the reference potential error caused by nonspecific adsorption in the actual sample test, thereby improving the test accuracy. In addition, the pH sensing channel is designed in the potential sensing array to calibrate the change of the response slope of the target sensing electrode channel to the target object caused by pH, so that the pH sensor is suitable for testing body fluid with wide pH range and has wide application in the aspect of home health monitoring. The innovation of the present invention is as follows.
Drawings
FIG. 1 is a scanning image of a transmission electron microscope of a PtAu/CNTs nanoenzyme glucose sensing material and a PBA/CNTs reference potential calibration material in example 1 of the present invention;
FIG. 2 is a flow chart showing the preparation of a glucose potential sensor array according to example 1 of the present invention;
FIG. 3 is a schematic diagram of a reference potential calibration performance test of a glucose potential sensing array in example 1 of the present invention;
FIG. 4 is a schematic diagram of a response slope calibration test of a glucose potential sensor array in example 1 of the present invention;
FIG. 5 is a graph showing the potential response and the standard operating curve of the glucose potential sensing array in example 1 of the present invention.
Detailed Description
The invention is described in further detail below with reference to specific embodiments and the attached drawings.
Example 1
1, preparing a PtAu/CNTs nano enzyme glucose sensing material:
dissolving 1mL of chloroauric acid aqueous solution and 0.8mL of chloroplatinic acid aqueous solution in deionized water at room temperature, then placing reactants in an ice water bath, dropwise adding 1mL of sodium borohydride aqueous solution with the concentration of 15mg/mL, and continuously reacting for 1 h; and finally, adding 10mg of aminated carbon nano tube, continuously reacting for 2h, and removing residual reactants of the final product through centrifugal washing to obtain the PtAu/CNTs nano enzyme. A scanning image of the PtAu/CNTs nanoenzyme glucose sensing material by a transmission electron microscope is shown in FIG. 1A. As shown in fig. 1A, the PtAu nanoparticles have a particle size of about 8nm and are uniformly distributed on the surface of the carbon nanotubes.
Preparing a PBA/CNTs reference potential calibration material:
30mg of 4-carboxyphenylboronic acid is dispersed in 20mL of DMSO, and after stirring, 3mg of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 2mg of N-hydroxysuccinimide are added. After the mixture was reacted at 45 ℃ for 12 hours, 20mg of aminated carbon nanotube was added to the mixture, and the mixture was stirred for 2 hours. Finally, the product was washed with deionized water and dried in a vacuum oven. A transmission electron microscope scan of the PBA/CNTs reference potential calibration material is shown in FIG. 1B.
Preparing a pH selective membrane solution:
a pH selective membrane solution was prepared by uniformly mixing 2mg of trilaurylamine, 1.2mg of sodium tetrakis (3, 5-bis (trifluoromethyl) phenyl) borate, 32.5mg of polyvinyl chloride, 65.5mg of o-nitrooctyl ether, and 1mL of anhydrous tetrahydrofuran.
4. Preparation of polyvinyl butyral resin reference film solution:
a reference film solution of polyvinyl butyral resin was prepared by uniformly mixing 100mg of sodium chloride, 250mg of polyvinyl butyral resin and 2mL of anhydrous methanol.
5. Preparing a glucose potential sensing array for simultaneously calibrating the reference potential and the response slope:
and preparing a four-channel base paper electrode array. A four-channel base paper electrode array comprising one row of silver/silver chloride electrode channels and three rows of carbon electrode channels was prepared by screen printing technique on 0.5mm thick PP synthetic paper. The schematic diagram of the base paper electrode array is shown in fig. 2A, in which a USB interface 1, a circuit channel 2, a reference electrode sensing area 3 and a carbon electrode sensing area 4 form the base paper electrode array. The USB interface 1, the circuit channel 2 and the reference electrode sensing area 3 form a silver/silver chloride electrode channel; the USB interface 1, the circuit channel 2 and the carbon electrode sensing area 4 form a carbon electrode channel. The diameter of the electrode is 3 mm. The specific printing process is as follows: PP synthetic paper with the thickness of 0.5mm is selected as a base material of the electrode. Carbon ink as a conductive layer was printed at the USB interface 1, the circuit path 2, the reference electrode sensing area 3, and the carbon electrode sensing area 4, and dried in an oven at 100 ℃ for 20 minutes. The silver/silver chloride electrode is then prepared by printing a silver/silver chloride ink at the reference electrode sensing area 3. Finally, the insulating paste was printed on all PP synthetic paper except for the USB interface 1, the reference electrode sensing area 3, and the carbon electrode sensing area 4, and dried in an oven at 100 ℃ for 20 minutes.
And (3) modifying a four-channel base paper electrode array. And dripping 15 mu L of polyvinyl butyral resin reference film solution on a reference electrode sensing area 3 of the base paper electrode array to serve as a reference electrode channel. And dripping the PBA/CNTs reference potential calibration material with the concentration of 1mg/mL of 10 muL on a carbon electrode sensing area 4 of the base paper electrode array to serve as a reference potential calibration channel. And dropwise coating the 10 muL PtAu/CNTs nano enzyme glucose sensing material with the concentration of 1mg/mL on another carbon electrode sensing area 4 of the base paper electrode array to form a glucose sensing electrode channel. And dripping 20 mu L of pH selective membrane solution on another carbon electrode sensing area 4 of the base paper electrode array to form a pH sensing electrode channel. And then drying the substrate in vacuum at 37 ℃ for 5 hours to obtain a glucose potential sensing array which simultaneously calibrates the reference potential and the response slope, and putting the glucose potential sensing array into a refrigerator at 4 ℃ for standby. A schematic diagram of a glucose potential sensing array capable of calibrating a reference potential and a response slope at the same time is shown in fig. 2B, in which a USB interface 1, a circuit channel 2 and a reference electrode 5 constitute a reference electrode channel; the USB interface 1, the circuit channel 2 and the reference potential calibration electrode 6 form a reference potential calibration channel; the USB interface 1, the circuit channel 2 and the glucose sensing electrode 7 form a glucose sensing electrode channel; the USB interface 1, the circuit channel 2 and the pH sensing electrode 8 form a pH sensing electrode channel, and a glucose potential sensing array capable of calibrating a reference potential and a response slope simultaneously is formed together.
6. And (3) a reference potential calibration performance test of the glucose potential sensing array capable of calibrating the reference potential and the response slope at the same time:
the actual sample has a complex composition, and interferes with the reference potential of the sensor, resulting in measurement errors. Therefore, it is very necessary to automatically calibrate the reference potential during the detection. Designs in a glucose potential sensing array capable of calibrating a reference potential and a response slope simultaneouslyA reference potential calibration channel, so as to realize automatic calibration of the reference potential in the detection process. The design principle of the reference potential calibration channel is as follows: the reference potential calibration channel has no potential response to the target object, and simultaneously, the response of the reference potential calibration channel to the interference potential generated by nonspecific adsorption in the sample is consistent with that of the target sensing electrode. Urine contains primarily electrolyte ions, non-protein nitrides, and may also contain proteins. Therefore, the glucose sensing channel and the reference potential calibration channel in the glucose potential sensing array were tested for the component (K) contained in urine +，Na+，Ca2 +Urea, uric acid, creatinine, hemoglobin and globin G) were subjected to potential measurements, see fig. 3. The glucose potential sensing array is firstly inserted into 10mM phosphate buffer solution to measure the potential value, and then is respectively inserted into a glucose solution containing 20mM, 100mM potassium ions, 100mM sodium ions, 20mM calcium ions, 1mM uric acid, 0.1M urea, 5mM creatinine, 0.2mg/mL globulin G and 0.2mg/mL hemoglobin to measure the potential value. As can be seen from FIG. 3, the glucose sensing channel produced a significant potential change in the glucose solution, while the reference potential calibration channel did not. In addition, the glucose sensing channel and the reference potential calibration channel have small and substantially consistent changes in the potential of other components in the urine other than glucose, demonstrating that the reference potential of the glucose sensing channel can be calibrated by subtracting the potential value of the reference potential calibration channel from the potential value of the glucose sensing channel.
7. The response slope calibration of the glucose potential sensing array can simultaneously calibrate the reference potential and the response slope:
it is well known that pH influences the activity of enzymes. In actual sample testing, pH is dynamically changing, thus the effect of pH on the slope of the glucose sensing channel response needs to be calibrated. Since the pH of human urine varies in the range of 4-7.5, the response performance of the glucose sensing channel to glucose was tested at pH =3.98, pH =4.96, pH =6.08, and pH = 7.45. As shown in fig. 4A, the slope of the response of the glucose sensing channel to glucose increases with increasing pH, indicating that neutral and alkaline conditions are more favorable for catalysis by glucose. The intercept and slope of the glucose sensing channel are plotted against pH to give FIG. 4B, which shows that the intercept and slope of the glucose sensing channel are linear with pH as shown in FIG. 4B. Let a and b be calibration coefficients for the intercept and slope, respectively, of the glucose sensing channel. The formula is as follows:
wherein SlopepH=6.08，T=302.8KAnd InterceptpH=6.08, T=302.8KThe response slope and intercept of the glucose sensing channel are respectively the test conditions of T =302.8K and pH = 6.08; slopepH=real, T=302.8KAnd InterceptpH=real, T=302.8KThe slope and intercept of the glucose sensing channel are measured at the actual pH measurement under test conditions T =302.8K, respectively. Therefore, the real-time pH value can be tested through the pH sensing channel in the glucose potential sensing array, and the response slope of the glucose sensing channel is directly calibrated.
8. And (3) potential testing:
inserting a glucose potential sensing array capable of calibrating a reference potential and a response slope simultaneously into an artificial urine sample containing glucose with different concentrations, and testing the potential value of each channel by adopting an open-circuit potential method. The potential response of the glucose sensing channel after calibrating the reference potential and the response slope in 0.032mM to 20mM is shown in FIG. 5A, and the glucose sensing channel has good potential response to glucose in the artificial urine sample. The results of linear fitting of the logarithmic values of the glucose concentration and the potential of the glucose sensing channel with the calibrated reference potential and response slope are shown in FIG. 5B, where the glucose sensing channel with the calibrated reference potential and response slope is shown at 10-2M-10-4There is a good linear response to glucose over the range of M concentrations.
Claims (2)
1. A preparation method of a potential sensing array for calibrating a reference potential and a response slope simultaneously is characterized in that the potential sensing array is used for detecting the concentration of a disease indicator in body fluid, and the preparation method of the potential sensing array comprises the following steps:
(1) preparing a basic electrode array:
designing a basic electrode array pattern on a substrate material, and preparing a basic electrode array by a printing process; the basic electrode array comprises a row of silver/silver chloride electrode channels and at least three rows of carbon-based electrode channels; each column of electrode channels comprises a circuit channel, a USB interface and an electrode sensing area;
the printing process specifically comprises the following steps: printing carbon ink on a circuit channel, a USB interface and an electrode sensing area in each row of electrode channels, and drying in a drying oven at 50-200 ℃ for 3-10 h; then selecting an electrode channel printed with carbon ink, and printing silver/silver chloride ink on a sensing area of the electrode channel to obtain a silver/silver chloride electrode; finally, printing the insulating paste on all substrate materials except the USB interface and the electrode sensing area, and drying in an oven at 50-200 ℃ for 3-10 h;
the substrate material is synthetic paper, polyethylene terephthalate film or textile;
the printing process is screen printing, ink-jet printing or manual coating;
(2) modifying the basic electrode array to obtain a potential sensing array which simultaneously calibrates a reference potential and a response slope:
2-1, preparing a polyvinyl butyral resin reference film solution: uniformly mixing sodium chloride, polyvinyl butyral resin and anhydrous methanol according to the mass-volume ratio of 50-100 mg: 100-250 mg: 2mL to obtain a polyvinyl butyral resin reference membrane solution; dripping polyvinyl butyral resin reference film solution on the surface of a silver/silver chloride electrode of a basic electrode array to form a reference electrode channel; wherein the thickness of the reference film of the polyvinyl butyral resin is 5-50 mu m;
2-2, modifying the target sensing material on the surface of any column of carbon-based electrodes in the basic electrode array to form a target sensing electrode channel; modifying the surface of any column of unmodified carbon-based electrodes in the basic electrode array by using a reference potential calibration material to form a reference potential calibration channel; modifying the pH sensing material on the surface of any column of unmodified carbon-based electrodes in the basic electrode array to form a pH sensing electrode channel; vacuum drying at 10-37 deg.C for 2-10h to obtain a potential sensing array with calibration reference potential and response slope, and storing at 0-10 deg.C;
the target sensing material is biological enzyme or nano enzyme;
the reference potential calibration material is a biological enzyme or nano-enzyme without catalytic activity;
the pH sensing material is a pH ion selective membrane, and the preparation steps of the pH ion selective membrane solution comprise: uniformly mixing 2mg of trilaurylamine, 1.2mg of sodium tetrakis (3, 5-bis (trifluoromethyl) phenyl) borate, 32.5mg of polyvinyl chloride, 65.5mg of o-nitrooctyl ether and 1mL of anhydrous tetrahydrofuran to obtain a pH ion selective membrane solution;
the modification method is a dropping coating method or an electrochemical deposition method;
the pH sensing electrode channel is used for calibrating the change of the potential response slope of the target sensing electrode channel to the target object caused by pH, so that the pH sensing electrode channel is suitable for testing body fluid with wide pH range.
2. A potential sensing array for simultaneously calibrating a reference potential and a response slope, manufactured according to the manufacturing method of claim 1.
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