CN117654654A - Miniature integrated device and method for measuring electrolyte content of urine - Google Patents
Miniature integrated device and method for measuring electrolyte content of urine Download PDFInfo
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- CN117654654A CN117654654A CN202311641905.2A CN202311641905A CN117654654A CN 117654654 A CN117654654 A CN 117654654A CN 202311641905 A CN202311641905 A CN 202311641905A CN 117654654 A CN117654654 A CN 117654654A
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- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention belongs to the technical field of urine electrolyte content detection, and particularly relates to a miniature integrated device and a miniature integrated method for measuring the urine electrolyte content; the silicon layer substrate is provided with a first electrode, and the third chamber layer is provided with an internal reference liquid pool and a third micro-flow channel; a sample pool and a second micro-flow channel are arranged in the second chamber layer; the silicon-based electrode layer is provided with a second electrode and a communication pool; a first microfluidic channel is arranged in the first chamber layer; the cover plate is provided with a potential measuring instrument; the whole device is provided with a first injection hole, a second injection hole, a third injection hole, a fourth injection hole and a fifth injection hole; the invention is based on the silicon microetching technology and combines the ion selective electrode detection method principle, can obtain the bioelectrolyte detection instrument which has lower price and is easy to carry through the miniaturized electrode and the electrolyte chamber, and can further increase the ion types which can be detected by the product by increasing the number of the internal reference liquid containers.
Description
Technical Field
The invention belongs to the technical field of urine electrolyte content detection, and particularly relates to a miniature integrated device and method for measuring the urine electrolyte content.
Background
The detection of electrolytes such as Na+, K+, ca2+, cl-and the like in urine is a common detection item and has great significance in clinic. Among them, abnormality of urinary sodium concentration is closely related to various renal diseases such as acute tubular necrosis, adrenocortical hypofunction and the like. The potassium ion concentration is important for maintaining normal stress of cardiac muscle and neuromuscular. Too high urinary potassium concentration can cause pathological changes of the cardiovascular system, so that cardiac muscle is inhibited, cardiac muscle tension is reduced, arrhythmia occurs in serious patients, and too low urinary potassium concentration can cause serious symptoms of muscle weakness and arrhythmia. Meanwhile, the urinary sodium and urinary potassium levels are closely related to the blood pressure and arterial stiffness of patients with hypertension. Abnormal urinary calcium concentration may cause serious diseases such as hypocalcemia and nephrotic syndrome of human body, and abnormal urinary chlorine concentration may cause epilepsy, syncope, convulsion and other symptoms of patients. Therefore, in order to adapt to the mobile rapid detection scenes of emergency treatment, home medical treatment, field rescue and the like, the method is very necessary for accurately, conveniently and rapidly detecting the human urine electrolyte.
Microfluidic technology is a technology for controlling fluids in a micrometer scale space, and early researches mainly focused on chemical electrophoresis analysis of continuous phase fluids, so that the technology is also called a micro total analysis system and a lab-on-a-chip. One of the important features of microfluidic is the unique fluidic properties in a microscale environment, such as laminar flow and droplets. By means of the above fluid phenomenon, the micro-fluidic technology can realize micro-processing and micro-operation which are difficult to be completed by the conventional method.
The electrolyte detection commonly used in the current technical background is based on electrochemical electrolyte analysis, and the target ion concentration is measured by using a larger measuring instrument through an ion selective electrode method, wherein the measuring principle is that a primary battery is used for measuring the membrane potential between a solution with known ion concentration and a solution to be measured, and then the potential is used for calculating the target ion concentration, wherein the basis is a Nernst equation. The other measuring methods include microelectrode array chip detection (preparing sensing microelectrode by combining PMMA hydrogel with high conductivity and metallographic phase with catalytic activity, realizing detection of trace potassium ions in human body electrolyte), ion selective electrode detection test paper (by preparing test strip electrode and integrating reference solution in the test paper, measuring fingertip blood electrolyte) and the like.
The existing urine electrolyte detection is often provided with a large instrument, the convenience degree is low, the cost is high, the requirement for rapidly detecting the concentration of the electrolyte in urine cannot be met, the operation is complex, and professional medical staff is required to assist in measurement. At present, a method for detecting the concentration of body fluid electrolyte, such as microelectrode array chip detection, ion selective electrode detection test paper and the like, has a certain degree of convenience, and most of the methods aim at detecting the electrolyte in human blood, and the accuracy and stability of detection are required to be improved.
Disclosure of Invention
In order to overcome the defects, the invention provides a miniature integrated device and a miniature integrated method for measuring the electrolyte content of urine, which are based on a silicon microetching technology and combined with an ion selective electrode detection method principle, so that the volume is reduced and the performance is improved to meet higher use requirements; by miniaturizing the electrodes and the electrolyte chambers, a low-priced, portable bioelectrolyte detecting instrument can be obtained, and the number of internal reference liquid containers can be increased, thereby further increasing the ion species that the product can detect.
A miniature integrated device for measuring the electrolyte content of urine comprises a silicon layer substrate 1, a third chamber layer 2, a sensitive film layer 3, a second chamber layer 4, a salt bridge 5, a silicon-based electrode layer 6, a first chamber layer 7 and a cover plate 8 which are sequentially connected together from bottom to top;
the silicon layer substrate 1 is integrated with a plurality of first electrodes 9, a plurality of internal reference liquid pools 10 which are in one-to-one correspondence with the first electrodes 9 are arranged in the third cavity layer 2, a third microfluidic channel 11 communicated with the internal reference liquid pools 10 is arranged on the third cavity layer 2 at the front end of the internal reference liquid pools 10, and third microfluidic channels 11 communicated with the corresponding internal reference liquid pools 10 are also respectively arranged on the third cavity layer 2 at the outer sides of the internal reference liquid pools 10 at the left side and the right side;
A sample cell 12 is arranged in the second chamber layer 4, and second microfluidic channels 13 communicated with the sample cell 12 are respectively arranged in the second chamber layers 4 at two sides of the sample cell 12;
the silicon-based electrode layer 6 is provided with a second electrode 14 and a communication pool 15;
a reference cell 21 and a first microfluidic channel 22 communicated with the reference cell 21 are arranged in the first chamber layer 7;
the top of the cover plate 8 is provided with a potential measuring instrument 23;
the miniature integrated device is provided with a first injection hole 16 penetrating through the cover plate 8; a second injection hole 17 penetrating the cover plate 8, the first chamber layer 7, the silicon-based electrode layer 6, and the salt bridge 5 in this order; the third injection hole 18, the fourth injection hole 19 and the fifth injection hole 20 sequentially penetrate through the cover plate 8, the first chamber layer 7, the silicon-based electrode layer 6, the salt bridge 5, the second chamber layer 4 and the sensitive film layer 3;
the first injection hole 16 communicates with the reference cell 21 of the first chamber layer 7 through a first microfluidic channel 22; the second injection hole 17 communicates with the sample cell 12 of the second chamber layer 4 through the second microfluidic channel 13; the third injection hole 18, the fourth injection hole 19 and the fifth injection hole 20 are respectively communicated with the three internal reference cells 10 of the third chamber layer 2 through corresponding third micro-flow channels 11;
the sensitive film layer 3 is formed by connecting three sensitive films with selective responses to sodium ions, potassium ions and calcium ions through a glue adhesive, and the three sensitive films are sequentially covered on the corresponding areas of the three internal reference liquid pools 10.
The left side and the right side of the micro-integrated device are respectively provided with a first injection hole 16, a second injection hole 17 and a third injection hole 18, the front end and the rear end of the micro-integrated device are respectively provided with a fourth injection hole 19, and the fifth injection hole 20 is arranged at the front end of the micro-integrated device.
The first electrode 9 and the second electrode 14 are Ag/AgCl electrodes, and the second electrode 14 is a common electrode of all the first electrodes 9.
The salt bridge 5 is a carbon nanotube bundle film.
The sensitive film forming the sensitive film layer 3 is prepared by spin-coating a sensitive film solution formed by mixing a PVC matrix, a plasticizer, a neutral complexing carrier, an ion additive and a solvent on a wafer substrate.
The plasticizer is dibutyl, dioctyl or dinonyl phthalate, dibutyl phthalate of phthalic acid, oxalic acid or sebacic acid;
when the sensitive film has selective response to sodium ions, the neutral complexing carrier is a Na ion carrier, and is specifically one or more of double crown ether, 4 '-benzo-12-crown-4, tetraphenyl 24-crown ether-8 and 1, 1-tris [1' - (2 '-oxa-4' -oxo-5 '-aza-5' -methyl) dodecyl ] propane;
when the sensitive film has selective response to potassium ions, the neutral complexing carrier is a potassium ion carrier, and specifically is one or more of macrocyclic antibiotics, di-tert-butyl or dimethyl-dibenzo-crown-10 and 1,1' -binaphthyl-20-crown-6;
When the sensitive film has selective response to calcium ions, the neutral complexing carrier is a calcium ion carrier, and is specifically one or more of N, N-di [ (11-ethyl) undecyl ] -N, N' -4, 5-tetramethyl-3, 6-dioxaoctanediamide, 1-tris (N-methyl-N-phenylaminocarbomethoxymethyl) propane and 1, 1-tris (N-methyl-N-phenylaminocarbomethoxymethyl) propane;
the ion additive is one or two of sodium tetraphenyl borate and potassium tetraphenyl borate;
the solvent is one or two of tetrahydrofuran and cyclohexanone.
A method for measuring electrolyte content in urine by using a miniature integrated device for measuring electrolyte content in urine, comprising the following steps:
step one, all electrodes of the device are connected with a potential measuring instrument 23 on a cover plate 8 through leads respectively;
step two, injecting a reference solution into the device through the first injection hole 16, enabling the reference solution to enter the reference cell 21 through the first injection hole 16 and the first microfluidic channel 22 in sequence, and stopping injection when the solution in the first injection hole 16 at the other side of the cover plate 8 is exposed;
step three, injecting the inner reference solution into the device through a third injection hole 18, a fourth injection hole 19 and a fifth injection hole 20 respectively, and sequentially enabling the inner reference solution to enter the inner reference liquid pool 10 through a third micro-flow channel 11; stopping injection when the solution is exposed in the third injection hole 18, the fourth injection hole 19 and the fifth injection hole 20 on the other side of the cover plate 8;
Step four, injecting a urine sample into the device through the second injection hole 17, and enabling the urine sample to enter the sample tank 12 through the second injection hole 17 and the second micro-flow channel 13 in sequence; stopping injection when the solution is exposed in the second injection hole 17 on the other side of the cover plate 8;
and fifthly, after the reading of the potential measuring instrument 23 is stable, reading the values corresponding to the three first electrodes 9, wherein the three values are the potential values received by the potential measuring instrument 23, and the potential values are calculated by the built-in micro-processor and converted into the corresponding concentrations of sodium, potassium and calcium ions.
The internal reference solution is 10 -2 M NaCl solution, 10 -2 KCl solution of M, 10 -2 CaCl of M 2 A solution; the reference solution is a saturated KCl solution.
The manufacturing method of the miniature integrated device for measuring the electrolyte content of urine specifically comprises the following steps:
step one, integrating three first electrodes 9 on a silicon layer substrate 1 to obtain an indication electrode;
step two, silicon is taken, and a layer of SiO is deposited on the silicon 2 In SiO 2 Spin-coating photoresist on the surface, and etching away part of the photoresist by utilizing MEMS lithography technology to obtain a third chamber layer 2 structure plane mask; the planar mask covering part of the structure of the third chamber layer 2 is an internal reference liquid pool 10 and a third microfluidic channel 11 to be obtained;
Etching the third chamber layer 2 through HF, and enabling the planar mask of the structure of the third chamber layer 2 to be free of SiO 2 Etching to obtain a three-dimensional structure of the third chamber layer 2 preliminarily;
step four, removing the plane mask of the third chamber layer structure to obtain a final third chamber layer 2;
step five, manufacturing a second chamber layer 4, a silicon-based electrode layer 6, a first chamber layer 7 and a cover plate 8 by adopting the same method as the second to fourth steps;
step six, integrating a second electrode 14 on the silicon-based electrode layer 6 to obtain a reference electrode;
preparing three kinds of sensitive film solutions with selective responses to sodium, potassium and calcium ions respectively, spin-coating the sensitive film solutions on a wafer substrate to prepare corresponding sensitive films respectively, and connecting the three kinds of sensitive films through a glue adhesive to form a sensitive film layer 3;
step eight, bonding the silicon layer substrate 1, the third chamber layer 2, the sensitive film layer 3, the second chamber layer 4, the salt bridge 5, the silicon-based electrode layer 6, the first chamber layer 7 and the cover plate 8 through glue stacking from bottom to top.
The manufacturing method of the salt bridge 5 comprises the following steps: a thermal decoupling CVD method is adopted;
step one, flowing a mixture gas of 100sccm He and 400sccm H2 in a tubular furnace for 10 minutes while heating the furnace to 775 ℃;
Inserting the wafer into a furnace by using a magnetic coupling transfer arm, and then maintaining the same gas and flow rate as those in the first step, and maintaining the furnace at the temperature of 775 ℃ for 10 minutes;
changing the gas and the flow rate into a mixed gas of 100sccm of C2H4, 400sccm of He and 100sccm of H2 for 3min, and heating at 775 ℃; after growing carbon nanotubes on a wafer, keeping the same gas and flow, cooling the furnace to below 100 ℃, and finally purging with 1000sccm He for 5min to obtain a carbon capillary array;
step four, depositing a layer of SiO on the silicon substrate 2 In SiO 2 Spin-coating photoresist on the surface, etching part of the photoresist by using MEMS lithography technology to obtain a salt bridge 5 injection hole plane mask, covering the plane mask to obtain each injection hole, and etching SiO by HF 2 Depositing a layer, etching SiO not protected by photoresist 2 Part, obtain the three-dimensional structure of each injection hole, remove the photoresist mask, get the SiO of each injection hole 2 A hard mask;
step five, covering the SiO obtained in the step four on the carbon capillary array obtained in the step two 2 Hard materialMask, etching carbon capillary array by KOH with mass concentration of 35%, etching SiO 2 The carbon capillary array part not protected by the hard mask obtains the three-dimensional structure of each injection hole and removes SiO 2 The hard mask, a salt bridge 5 with individual injection holes is obtained.
In the seventh step, the preparation method of the sensitive film comprises the following steps:
firstly, preparing 20ml of selective membrane solution with selective responses to sodium, potassium and calcium ions respectively; wherein, the selective membrane solution of three ions uses one or two of cyclohexanone and tetrahydrofuran as mixed solvent; sodium tetraphenylborate and potassium tetraphenylborate as ion additives; polyvinyl chloride is used as a PVC matrix, and dioctyl sebacate is used as a plasticizer;
secondly, respectively dripping the prepared selective membrane solution on a wafer substrate, and spin-coating uniformly;
and thirdly, volatilizing the mixed solvent at room temperature to obtain a sensitive film, and cutting the sensitive film into a shape with proper size.
When the sodium ion selective membrane solution is prepared, tetraphenyl 24-crown ether-8 is selected as a sodium ion carrier, and the mass ratio of the ion additive, the ion carrier, the polyvinyl chloride and the dioctyl sebacate is 1.8:0.2:24:74;
when the potassium ion selective membrane solution is prepared, 1' -binaphthyl-20-crown-6 is selected as a carrier of potassium ions, and the mass ratio of an ion additive to the ion carrier to the polyvinyl chloride to the dioctyl sebacate is 1.8:0.2:33: 65.
When the calcium ion selective membrane solution is prepared, 1-tri (N-methyl-N-phenyl amino carbonyl methoxymethyl) propane is selected as a carrier of calcium ions, and an ionic additive, an ionic carrier, polyvinyl chloride and dioctyl sebacate are mixed according to the mass ratio of 1.8:0.2:32:66;
the mixed solvent is 20ml mixed solvent of cyclohexanone and tetrahydrofuran.
The invention has the beneficial effects that:
1. the MEMS silicon micro-etching technology is utilized, so that the volume of the detector is greatly reduced, and the detector can integrate the indicating electrode and the reference electrode to the maximum extent, thereby reducing the cost of the bioelectrolyte detecting instrument; the accurate detection result is obtained while the use amount of the detection liquid is reduced;
2. multiple bioelectrolytes can be detected simultaneously by adding an internal reference liquid chamber; specific ion concentrations can be detected by varying the sensitive membrane and the internal reference solution;
3. MEMS silicon micro-etching technology is mature at present, and the portability and integration degree of the MEMS silicon micro-etching technology can be greatly improved while the detection performance is ensured.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following description will briefly explain the drawings to be used in the description of the embodiments of the present invention, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to the contents of the embodiments of the present invention and these drawings without inventive effort for those skilled in the art.
Fig. 1 is a schematic view of the overall exploded structure of the present invention.
Fig. 2 is a schematic side view of the present invention.
FIG. 3 is a schematic view of a silicon layer substrate structure according to the present invention.
Fig. 4 is a schematic view of a third chamber layer structure according to the present invention.
FIG. 5 is a schematic view of a sensitive film structure according to the present invention.
Fig. 6 is a schematic view of a second chamber layer structure according to the present invention.
Fig. 7 is a schematic diagram of a salt bridge structure according to the present invention.
FIG. 8 is a schematic view of a silicon-based electrode layer structure according to the present invention.
Fig. 9 is a schematic view of a first chamber layer structure according to the present invention.
Fig. 10 is a schematic diagram of a cover plate structure according to the present invention.
Wherein: the substrate comprises a silicon layer substrate 1, a third chamber layer 2, a sensitive film layer 3, a second chamber layer 4, a salt bridge 5, a silicon-based electrode layer 6, a first chamber layer 7, a cover plate 8, a first electrode 9, an internal reference liquid pool 10, a third micro-flow channel 11, a sample pool 12, a second micro-flow channel 13, a second electrode 14, a communication pool 15, a first injection hole 16, a second injection hole 17, a third injection hole 18, a fourth injection hole 19, a fifth injection hole 20, a reference pool 21, a first micro-flow channel 22 and a potential measuring instrument 23.
Detailed Description
The invention is described in further detail below with reference to the drawings and examples. It is to be understood that the specific embodiments described herein are merely illustrative of the invention and are not limiting thereof. It should be further noted that, for convenience of description, only some, but not all of the structures related to the present invention are shown in the drawings.
Example 1
As shown in fig. 1-10, a micro integrated device for measuring the electrolyte content of urine comprises a silicon layer substrate 1, a third chamber layer 2, a sensitive film layer 3, a second chamber layer 4, a salt bridge 5, a silicon-based electrode layer 6, a first chamber layer 7 and a cover plate 8 which are sequentially connected together from bottom to top;
the silicon layer substrate 1 is integrated with a plurality of first electrodes 9, a plurality of internal reference liquid pools 10 which are in one-to-one correspondence with the first electrodes 9 are arranged in the third cavity layer 2, a third microfluidic channel 11 communicated with the internal reference liquid pools 10 is arranged on the third cavity layer 2 at the front end of the internal reference liquid pools 10, and third microfluidic channels 11 communicated with the corresponding internal reference liquid pools 10 are also respectively arranged on the third cavity layer 2 at the outer sides of the internal reference liquid pools 10 at the left side and the right side;
a sample cell 12 is arranged in the second chamber layer 4, and second microfluidic channels 13 communicated with the sample cell 12 are respectively arranged in the second chamber layers 4 at two sides of the sample cell 12;
the silicon-based electrode layer 6 is provided with a second electrode 14 and a communication pool 15;
a reference cell 21 and a first microfluidic channel 22 communicated with the reference cell 21 are arranged in the first chamber layer 7;
the top of the cover plate 8 is provided with a potential measuring instrument 23;
the miniature integrated device is provided with a first injection hole 16 penetrating through the cover plate 8; a second injection hole 17 penetrating the cover plate 8, the first chamber layer 7, the silicon-based electrode layer 6, and the salt bridge 5 in this order; the third injection hole 18, the fourth injection hole 19 and the fifth injection hole 20 sequentially penetrate through the cover plate 8, the first chamber layer 7, the silicon-based electrode layer 6, the salt bridge 5, the second chamber layer 4 and the sensitive film layer 3;
The first injection hole 16 communicates with the reference cell 21 of the first chamber layer 7 through a first microfluidic channel 22; the second injection hole 17 communicates with the sample cell 12 of the second chamber layer 4 through the second microfluidic channel 13; the third injection hole 18, the fourth injection hole 19 and the fifth injection hole 20 are respectively communicated with the three internal reference cells 10 of the third chamber layer 2 through corresponding third micro-flow channels 11;
the sensitive film layer 3 is formed by connecting three sensitive films with selective responses to sodium ions, potassium ions and calcium ions through a glue adhesive, and the three sensitive films are sequentially covered on the corresponding areas of the three internal reference liquid pools 10.
The left side and the right side of the micro-integrated device are respectively provided with a first injection hole 16, a second injection hole 17 and a third injection hole 18, the front end and the rear end of the micro-integrated device are respectively provided with a fourth injection hole 19, and the fifth injection hole 20 is arranged at the front end of the micro-integrated device.
The first electrode 9 and the second electrode 14 are Ag/AgCl electrodes, and the second electrode 14 is a common electrode of all the first electrodes 9.
The salt bridge 5 is a carbon nanotube bundle film, which is a porous glass or polymer frit, a polymer film, carbon capillaries (bundles) (also called carbon nanotube bundle film), preferably a carbon nanotube bundle film, which is a carbon capillary array with a high aspect ratio.
The sensitive film forming the sensitive film layer 3 is prepared by spin-coating a sensitive film solution formed by mixing a PVC matrix, a plasticizer, a neutral complexing carrier, an ion additive and a solvent on a wafer substrate.
The plasticizer is dibutyl, dioctyl or dinonyl phthalate, dibutyl phthalate of phthalic acid, oxalic acid or sebacic acid;
when the sensitive film has selective response to sodium ions, the neutral complexing carrier is a Na ion carrier, and is specifically one or more of double crown ether, 4 '-benzo-12-crown-4, tetraphenyl 24-crown ether-8 and 1, 1-tris [1' - (2 '-oxa-4' -oxo-5 '-aza-5' -methyl) dodecyl ] propane;
when the sensitive film has selective response to potassium ions, the neutral complexing carrier is a potassium ion carrier, and specifically is one or more of macrocyclic antibiotics, di-tert-butyl or dimethyl-dibenzo-crown-10 and 1,1' -binaphthyl-20-crown-6;
when the sensitive film has selective response to calcium ions, the neutral complexing carrier is a calcium ion carrier, and is specifically one or more of N, N-di [ (11-ethyl) undecyl ] -N, N' -4, 5-tetramethyl-3, 6-dioxaoctanediamide, 1-tris (N-methyl-N-phenylaminocarbomethoxymethyl) propane and 1, 1-tris (N-methyl-N-phenylaminocarbomethoxymethyl) propane;
The ion additive is one or two of sodium tetraphenyl borate and potassium tetraphenyl borate;
the solvent is one or two of tetrahydrofuran and cyclohexanone.
A method for measuring electrolyte content in urine by using a miniature integrated device for measuring electrolyte content in urine, comprising the following steps:
step one, all electrodes of the device are connected with a potential measuring instrument 23 on a cover plate 8 through leads respectively;
step two, injecting a reference solution into the device through the first injection hole 16 on the left side (or the right side), enabling the reference solution to sequentially enter the reference cell 21 through the first injection hole 16 and the first microfluidic channel 22 on the left side (or the right side), and stopping injecting when the solution is slightly exposed in the first injection hole 16 on the other side of the cover plate 8;
step three, injecting the inner reference solution into the device through a third injection hole 18 on the left side (or the right side), a fourth injection hole 19 and a fifth injection hole 20 respectively, and enabling the inner reference solution to enter the inner reference liquid pool 10 through a third micro-flow channel 11 on the left side (or the right side) of the third injection hole 18 (the fourth injection hole 19 and the fifth injection hole 20) in sequence; stopping injection when solutions are slightly exposed in the third injection hole 18, the fourth injection hole 19 and the fifth injection hole 20 on the other side of the cover plate 8;
Step four, injecting a urine sample into the device through the second injection hole 17 on the left side (or the right side), and sequentially enabling the urine sample to enter the sample tank 12 through the second injection hole 17 and the second microfluidic channel 13 on the left side (or the right side); stopping injection when the solution is slightly exposed in the second injection hole 17 on the other side of the cover plate 8;
and fifthly, after the reading of the potential measuring instrument 23 is stable, reading the values corresponding to the three first electrodes 9, wherein the three values are the concentrations of the corresponding sodium, potassium and calcium ions in urine, which are calculated and converted by the built-in micro-processor from the potential values received by the potential measuring instrument 23.
The manufacturing method of the miniature integrated device for measuring the electrolyte content of urine specifically comprises the following steps:
step one, integrating three first electrodes 9 on a silicon layer substrate 1 by a chemical method, an electrochemical method or a screen printing method to obtain an indication electrode;
step two, silicon is taken, and a layer of SiO is deposited on the silicon 2 In SiO 2 Spin-coating photoresist on the surface, and etching away part of the photoresist by utilizing MEMS lithography technology to obtain a third chamber layer 2 structure plane mask; the planar mask covering part of the structure of the third chamber layer 2 is an internal reference liquid pool 10 and a third microfluidic channel 11 to be obtained;
Etching the third chamber layer 2 through HF, and enabling the planar mask of the structure of the third chamber layer 2 to be free of SiO 2 Etching to obtain a three-dimensional structure of the third chamber layer 2 preliminarily;
step four, removing the plane mask of the third chamber layer structure to obtain the three-dimensional structure of the final third chamber layer 2, namely an internal reference liquid pool 10 and a third micro-flow channel 11;
step five, manufacturing a second chamber layer 4, a silicon-based electrode layer 6, a first chamber layer 7 and a cover plate 8 by adopting the same method as the second to fourth steps;
the manufacturing method of the second chamber layer 4 comprises the following steps: depositing a layer of SiO on the silicon substrate 2 In SiO 2 Spin-coating photoresist on the surface, and etching away part of the photoresist by utilizing MEMS lithography technology to obtain a planar mask of the structure of the second chamber layer 4; the planar mask covers the parts of the sample cell 12, the second microfluidic channel 13 and the corresponding injection holes to be obtained, the second chamber layer 4 is etched by HF etching, the etching being performedPhotoresist unprotected SiO 2 A part for obtaining a three-dimensional structure of the sample cell 12, the second microfluidic channel 13 and the corresponding injection hole; removing the photoresist mask, namely the second cavity layer 4 structure plane mask, to obtain a sample cell 12, a second micro-flow channel 13 and a corresponding injection hole;
the manufacturing method of the silicon-based electrode layer 6 comprises the following steps: depositing a layer of SiO on the silicon substrate 2 In SiO 2 Spin-coating photoresist on the surface, and etching away part of the photoresist by utilizing MEMS photoetching technology to obtain a silicon-based electrode layer 6 structure plane mask; the planar mask covers the portions of the via hole 15 and the corresponding injection hole to be obtained, and the silicon-based electrode layer 6 is etched by HF etching, and SiO not protected by photoresist is etched 2 A part for obtaining a three-dimensional structure of the communication pool 15 and the corresponding injection hole; removing the photoresist mask to obtain a communication pool 15 and a corresponding injection hole;
the manufacturing method of the first chamber layer 7 comprises the following steps: depositing a layer of SiO on the silicon substrate 2 In SiO 2 Spin-coating photoresist on the surface, and etching away part of the photoresist by utilizing MEMS lithography technology to obtain a planar mask of the structure of the first chamber layer 7; the planar mask covers the portions of the reference cell 21, the first microfluidic channel 22 and the corresponding injection holes to be obtained, etching the first chamber layer 7 by HF etching, etching the SiO not protected by the photoresist 2 In part, a three-dimensional structure of the reference cell 21, the first microfluidic channel 22 and the corresponding injection hole is obtained; removing the photoresist mask to obtain a reference cell 21, a first microfluidic channel 22 and corresponding injection holes;
the manufacturing method of the cover plate 8 comprises the following steps: depositing a layer of SiO on the silicon substrate 2 In SiO 2 Spin-coating photoresist on the surface, etching part of the photoresist by using MEMS (micro electro mechanical System) photoetching technology to obtain a planar mask of the structure of the cover plate 8, wherein the coverage part of the planar mask is all the injection holes to be obtained, etching the cover plate 8 by HF (high-frequency) etching, and etching SiO (silicon dioxide) which is not protected by the photoresist 2 A part, obtaining a three-dimensional structure of each injection hole, and removing the photoresist mask to obtain each injection hole;
step six, integrating a second electrode 14 on the silicon-based electrode layer 6 by a chemical, electrochemical or screen printing method to obtain a reference electrode;
preparing three kinds of sensitive film solutions with selective responses to sodium, potassium and calcium ions respectively, spin-coating the sensitive film solutions on a wafer substrate to prepare corresponding sensitive films respectively, and connecting the three kinds of sensitive films through a glue adhesive to form a sensitive film layer 3; the three sensitive films are separated when being manufactured, and are connected together through an adhesive (glue), the connected large films are the sensitive film layers 3, and when the three sensitive films are stacked on the third chamber layer 2, the three sensitive films just cover the corresponding inner reference liquid pools 10 on the third chamber layer 2 respectively;
step eight, bonding the silicon layer substrate 1, the third chamber layer 2, the sensitive film layer 3, the second chamber layer 4, the salt bridge 5, the silicon-based electrode layer 6, the first chamber layer 7 and the cover plate 8 through glue stacking from bottom to top.
The manufacturing method of the salt bridge 5 comprises the following steps: a thermal decoupling CVD method is adopted;
step 1, flowing a mixture gas of 100sccm He and 400sccm H2 in a tubular furnace for 10 minutes while heating the furnace to 775 ℃;
Step 2, inserting the wafer into a furnace by using a magnetic coupling transfer arm, and then maintaining the same gas and flow rate as those in the step 1, and maintaining the furnace at the temperature of 775 ℃ for 10 minutes;
step 3, changing the gas and flow rate into 100sccm of C2H4, 400sccm He and 100sccm H2 mixed gas for 3min, and heating at 775 ℃; after growing carbon nanotubes on a wafer, keeping the same gas and flow, cooling the furnace to below 100 ℃, and finally purging with 1000sccm He for 5min to obtain a carbon capillary array;
step 4, depositing a layer of SiO on the silicon substrate 2 In SiO 2 Spin-coating photoresist on the surface, etching part of the photoresist by using MEMS lithography technology to obtain a salt bridge 5 injection hole plane mask, covering the plane mask to obtain each injection hole, and etching SiO by HF 2 Depositing a layer, etching SiO not protected by photoresist 2 Part, obtain the three-dimensional structure of each injection hole, remove the photoresist mask, get the SiO of each injection hole 2 A hard mask;
step 5, carbon obtained in step 2Coating the capillary array with SiO obtained in step 4 2 Hard mask, etching by KOH etching carbon capillary array with mass concentration of 35%, etching by SiO 2 The carbon capillary array part not protected by the hard mask obtains the three-dimensional structure of each injection hole and removes SiO 2 The hard mask, a salt bridge 5 with individual injection holes is obtained.
The internal reference solution is 10 -2 M NaCl solution, 10 -2 KCl solution of M, 10 -2 CaCl of M 2 A solution.
The reference solution is a saturated KCl solution.
In the seventh step, the preparation method of the sensitive film comprises the following steps:
firstly, preparing 20ml of selective membrane solution with selective responses to sodium, potassium and calcium ions respectively; wherein, the selective membrane solution of three ions uses one or two of cyclohexanone and tetrahydrofuran as mixed solvent; sodium tetraphenylborate and potassium tetraphenylborate as ion additives; polyvinyl chloride is used as a PVC matrix, and dioctyl sebacate is used as a plasticizer;
secondly, respectively dripping the prepared selective membrane solution on a wafer substrate, and spin-coating uniformly;
and thirdly, volatilizing the mixed solvent at room temperature to obtain a sensitive film, and cutting the sensitive film into a shape with proper size.
When the sodium ion selective membrane solution is prepared, tetraphenyl 24-crown ether-8 is selected as a sodium ion carrier, and the mass ratio of the ion additive, the ion carrier, the polyvinyl chloride and the dioctyl sebacate is 1.8:0.2:24:74;
when the potassium ion selective membrane solution is prepared, 1' -binaphthyl-20-crown-6 is selected as a carrier of potassium ions, and the mass ratio of an ion additive to the ion carrier to the polyvinyl chloride to the dioctyl sebacate is 1.8:0.2:33: 65.
When the calcium ion selective membrane solution is prepared, 1-tri (N-methyl-N-phenyl amino carbonyl methoxymethyl) propane is selected as a carrier of calcium ions, and an ionic additive, an ionic carrier, polyvinyl chloride and dioctyl sebacate are mixed according to the mass ratio of 1.8:0.2:32:66;
the mixed solvent is 20ml mixed solvent of cyclohexanone and tetrahydrofuran.
Example 2
A miniature integrated device for measuring the electrolyte content of urine comprises a silicon layer substrate 1 arranged below, wherein three first electrodes 9 of Ag/AgCl are integrated on the silicon layer substrate 1; the inner reference liquid pool 10 in the attached third chamber layer 2 is used for containing the inner reference liquid, the inner reference liquid enters the inner reference liquid pool 10 through the fourth injection hole 19 and then enters the third microfluidic channel 11, and the inner reference liquid pool 10 is filled under injection pressure; three sensitive films which have selective responses to sodium, potassium and calcium ions are adhered on the third chamber layer 2; the sensitive film layer 3 is connected with a second chamber layer 4, wherein a sample cell 12 holds sample solution entering through a second injection hole 16 and a microfluidic channel 13; the salt bridge 5 is connected and is a carbon nano tube bundle film, has lower impedance and is not easy to pollute the sample liquid; a silicon-based electrode layer 6 is connected above the salt bridge 5, and a second electrode 14 and a communication pool 15 are integrated on the silicon substrate, and the communication pool 15 enables the reference liquid in the first chamber layer 7 and the sample solution in the sample pool 12 to exchange ions, so that a circuit is conducted; the silicon-based electrode layer 6 is connected with the first chamber layer 7, wherein a reference cell 21 is used for containing reference liquid entering through the second injection hole 16 and the microfluidic channel 22; the uppermost layer is capped with a cover plate 8 etched from a silicon substrate. A potentiometric device 23 is integrated into the cover plate 8 for measuring the ionic activity of the sample solution.
A miniature integrated device for measuring the electrolyte content of urine. Structure referring to fig. 1, the structure includes a silicon layer substrate 1 disposed below, and three Ag/AgCl first electrodes 9 are integrated on the silicon layer substrate 1; the inner reference liquid pool 10 in the attached third chamber layer 2 is used for containing the inner reference liquid, the inner reference liquid enters the third microfluidic channel 11 through the injection hole 20, and the inner reference liquid pool 10 is filled under injection pressure; three sensitive films are adhered on the chamber layer 2, and have selective responses to sodium, potassium and calcium ions respectively; the sensitive film layer 3 is connected with the second chamber layer 4, and the sample pool 12 contains sample solution entering through the second injection hole 17 and the second micro-flow channel 13; the salt bridge 5 is connected and is a carbon nano tube bundle film, has lower impedance and is not easy to pollute the sample liquid; above the salt bridge 5A silicon-based electrode layer 6, which integrates an Ag/AgCl second electrode 14 and a communication cell 15 on a silicon substrate, and the communication cell 15 enables the reference liquid in the first chamber layer 7 and the sample solution in the sample cell 12 to exchange ions, thereby conducting a circuit; the silicon-based electrode layer 6 is connected with a first chamber layer 7, wherein a reference pool 21 is used for containing reference liquid entering through a first injection hole 16 and a first micro-flow channel 22; the uppermost layer is capped with a cover plate 8 etched from a silicon substrate. A potentiometric device 23 is integrated into the cover plate 8 for measuring the ionic activity of the sample solution. The whole structure forms a battery system, which can be expressed as Ag|AgCl(s), KCl (saturation) |0.1mol/L LiAc|sample liquid||sensitive film|0.01 mol/L Cl - AgCl(s) |Ag. The silicon layer substrate 1, the cavity and the sensitive film layer 3 form an indicating electrode, and the cover plate 8, the cavity, the silicon-based electrode layer 6 and the salt bridge 5 form a reference electrode. The whole structure size is 2cm x 8cm x 1cm.
Example 1: a method for patterning and etching a third chamber layer is provided
1. Growing a layer of SiO with the thickness of 1 mm on the surface of the cleaned silicon by PECVD 2 ;
2. The photoresist was coated on the sample surface using spin coating. The specific operation is as follows: the silicon wafer is vacuum adsorbed on a centrifugal type spin coater to rotate at high speed, and photoresist dropped on the surface of the silicon wafer is uniformly coated.
3. And heating to evaporate partial solvent of the photoresist to primarily cure the photoresist layer.
4. And (3) registering the mask plate and the silicon wafer alignment mark, exposing the photoresist to change the structure of a part of the region, and transferring the pattern of the third chamber layer structure.
5. And (3) placing the silicon wafer in a developing solution to dissolve and remove the irradiated part, thereby obtaining the planar mask of the third chamber layer structure.
6. SiO was treated with 40% strength HF solution 2 Wet etching is carried out on the layer to obtain a three-dimensional structure of the third chamber layer;
the main principle of the development method of the MEMS silicon microetching-based miniature integrated bioelectrolyte detector is an ion selective electrode method in a potentiometric analysis method, which is commonly used for detecting the concentration of specific ions in a solution. And by combining with the MEMS silicon micro-etching technology, the integration level of the detection equipment is improved, and the ultra-microminiaturization of the equipment is realized. The invention can selectively increase or decrease the internal reference cell and the corresponding sensitive membrane according to the bioelectrolyte detection scene of the specific medical scene to detect the specific bioelectrolytes. Etching each cavity layer by MEMS silicon micro-etching technology, bonding each electrode layer, the sensitive film and the salt bridge, and finally realizing the manufacture of the ion selective electrode method detector on the micrometer scale.
The preferred embodiments of the present invention have been described in detail above with reference to the accompanying drawings, but the scope of the present invention is not limited to the specific details of the above embodiments, and within the scope of the technical concept of the present invention, any person skilled in the art may apply equivalent substitutions or alterations to the technical solution according to the present invention and the inventive concept thereof within the scope of the technical concept of the present invention, and these simple modifications are all within the scope of the present invention.
In addition, the specific features described in the above embodiments may be combined in any suitable manner, and in order to avoid unnecessary repetition, various possible combinations are not described further.
Moreover, any combination of the various embodiments of the invention can be made without departing from the spirit of the invention, which should also be considered as disclosed herein.
Claims (10)
1. The miniature integrated device for measuring the electrolyte content of urine is characterized by comprising a silicon layer substrate (1), a third chamber layer (2), a sensitive film layer (3), a second chamber layer (4), a salt bridge (5), a silicon-based electrode layer (6), a first chamber layer (7) and a cover plate (8) which are sequentially connected from bottom to top;
The silicon layer substrate (1) is integrated with a plurality of first electrodes (9), a plurality of internal reference liquid pools (10) which are in one-to-one correspondence with the first electrodes (9) are arranged in the third chamber layer (2), a third microfluidic channel (11) communicated with the internal reference liquid pools (10) is arranged on the third chamber layer (2) at the front end of the internal reference liquid pools (10), and third microfluidic channels (11) communicated with the corresponding internal reference liquid pools (10) are also respectively arranged on the third chamber layer (2) at the outer sides of the internal reference liquid pools (10) at the left side and the right side;
a sample pool (12) is arranged in the second chamber layer (4), and second microfluidic channels (13) communicated with the sample pool (12) are respectively arranged in the second chamber layers (4) at two sides of the sample pool (12);
a second electrode (14) and a communication pool (15) are arranged on the silicon-based electrode layer (6);
a reference cell (21) and a first microfluidic channel (22) communicated with the reference cell are arranged in the first chamber layer (7);
the top of the cover plate (8) is provided with a potential measuring instrument (23);
the miniature integrated device is provided with a first injection hole (16) penetrating through the cover plate (8); a second injection hole (17) penetrating through the cover plate (8), the first chamber layer (7), the silicon-based electrode layer (6) and the salt bridge (5) in sequence; a third injection hole (18), a fourth injection hole (19) and a fifth injection hole (20) which sequentially penetrate through the cover plate (8), the first chamber layer (7), the silicon-based electrode layer (6), the salt bridge (5), the second chamber layer (4) and the sensitive film layer (3);
The first injection hole (16) is communicated with a reference pool (21) of the first chamber layer (7) through a first micro-flow channel (22); the second injection hole (17) is communicated with the sample pool (12) of the second chamber layer (4) through a second micro-flow channel (13); the third injection hole (18), the fourth injection hole (19) and the fifth injection hole (20) are respectively communicated with three inner reference cells (10) of the third chamber layer (2) through corresponding third micro-flow channels (11);
the sensitive film layer (3) is formed by connecting three sensitive films with selective responses to sodium ions, potassium ions and calcium ions through a glue adhesive, and the three sensitive films are sequentially covered on corresponding areas of the three internal reference liquid pools (10).
2. The micro-integrated device for measuring the electrolyte content of urine according to claim 1, wherein the left side and the right side of the micro-integrated device are respectively provided with a first injection hole (16), a second injection hole (17) and a third injection hole (18), the front end and the rear end of the micro-integrated device are respectively provided with a fourth injection hole (19), and the fifth injection hole (20) is arranged at the front end of the micro-integrated device.
3. A micro-integrated device for measuring the electrolyte content of urine according to claim 1, wherein the salt bridge (5) is a carbon nanotube bundle film.
4. The miniature integrated device for measuring the electrolyte content of urine according to claim 1, wherein the sensitive film forming the sensitive film layer (3) is prepared by spin-coating a sensitive film solution formed by mixing a PVC matrix, a plasticizer, a neutral complexing carrier, an ion additive and a solvent on a wafer substrate;
the plasticizer is dibutyl, dioctyl or dinonyl phthalate, dibutyl phthalate of phthalic acid, oxalic acid or sebacic acid;
when the sensitive film has selective response to sodium ions, the neutral complexing carrier is a Na ion carrier, and is specifically one or more of double crown ether, 4 '-benzo-12-crown-4, tetraphenyl 24-crown ether-8 and 1, 1-tris [1' - (2 '-oxa-4' -oxo-5 '-aza-5' -methyl) dodecyl ] propane;
when the sensitive film has selective response to potassium ions, the neutral complexing carrier is a potassium ion carrier, and specifically is one or more of macrocyclic antibiotics, di-tert-butyl or dimethyl-dibenzo-crown-10 and 1,1' -binaphthyl-20-crown-6;
when the sensitive film has selective response to calcium ions, the neutral complexing carrier is a calcium ion carrier, and is specifically one or more of N, N-di [ (11-ethyl) undecyl ] -N, N' -4, 5-tetramethyl-3, 6-dioxaoctanediamide, 1-tris (N-methyl-N-phenylaminocarbomethoxymethyl) propane and 1, 1-tris (N-methyl-N-phenylaminocarbomethoxymethyl) propane;
The ion additive is one or two of sodium tetraphenyl borate and potassium tetraphenyl borate;
the solvent is one or two of tetrahydrofuran and cyclohexanone.
5. A micro-integrated device for measuring the electrolyte content of urine according to claim 1, characterized in that the first electrode (9) and the second electrode (14) are both Ag/AgCl electrodes, and the second electrode (14) is a common electrode for all the first electrodes (9).
6. A method for measuring electrolyte content in urine using the micro-integrated device for measuring electrolyte content in urine according to any one of claims 1 to 5, comprising the following steps:
step one, all electrodes of the device are connected with a potential measuring instrument (23) on a cover plate (8) through leads respectively;
step two, injecting a reference solution into the device through a first injection hole (16), enabling the reference solution to sequentially enter a reference tank (21) through the first injection hole (16) and a first micro-flow channel (22), and stopping injecting when the solution is exposed in the first injection hole (16) at the other side of the cover plate (8);
injecting the inner reference solution into the device through a third injection hole (18), a fourth injection hole (19) and a fifth injection hole (20), and sequentially entering the inner reference solution into an inner reference solution pool (10) through a third micro-flow channel (11); stopping injection when solutions are exposed in the third injection hole (18), the fourth injection hole (19) and the fifth injection hole (20) on the other side of the cover plate (8);
Step four, injecting a urine sample into the device through a second injection hole (17), and enabling the urine sample to enter a sample pool (12) through the second injection hole (17) and a second micro-flow channel (13) in sequence; stopping injection when the solution is exposed in the second injection hole (17) on the other side of the cover plate (8);
and fifthly, after the reading of the potential measuring instrument (23) is stable, reading the corresponding values of three first electrodes (9), wherein the three values are the concentrations of the corresponding sodium, potassium and calcium ions, which are calculated by the built-in microprocessor and converted from the potential values received by the potential measuring instrument (23).
7. The method of measuring electrolyte content in urine as recited in claim 6, wherein the internal reference solution is 10- 2 M NaCl solution, 10- 2 KCl solution of M, 10- 2 CaCl of M 2 A solution; the reference solution is a saturated KCl solution.
8. A method for manufacturing a micro-integrated device for measuring the electrolyte content of urine according to any one of claims 1 to 5, comprising the steps of:
step one, integrating three first electrodes (9) on a silicon layer substrate (1) to obtain an indication electrode;
step two, silicon is taken, and a layer of SiO is deposited on the silicon 2 In SiO 2 Spin-coating photoresist on the surface, and etching away part of the photoresist by utilizing MEMS lithography technology to obtain a plane mask of the structure of the third chamber layer (2); the cover part of the third chamber layer (2) structure plane mask is an internal reference liquid pool (10) and a third micro-flow channel (11) to be obtained;
Etching the third chamber layer (2) through HF, and enabling the structure plane mask of the third chamber layer (2) to be in non-covered part of SiO 2 Etching to obtain a three-dimensional structure of the third chamber layer (2) preliminarily;
step four, removing the plane mask of the third chamber layer structure to obtain a final third chamber layer (2);
step five, manufacturing a second chamber layer (4), a silicon-based electrode layer (6), a first chamber layer (7) and a cover plate (8) by adopting the same method as the second to fourth steps;
step six, integrating a second electrode (14) on the silicon-based electrode layer (6) to obtain a reference electrode;
preparing three kinds of sensitive film solutions with selective responses to sodium, potassium and calcium ions respectively, spin-coating the sensitive film solutions on a wafer substrate to prepare corresponding sensitive films respectively, and connecting the three kinds of sensitive films through a glue adhesive to form a sensitive film layer (3);
step eight, bonding a silicon layer substrate (1), a third cavity layer (2), a sensitive film layer (3), a second cavity layer (4), a salt bridge (5), a silicon-based electrode layer (6), a first cavity layer (7) and a cover plate (8) through glue stacking.
9. A method for manufacturing a salt bridge (5) in a micro-integrated device for measuring the electrolyte content of urine according to any one of claims 1 to 5, characterized in that a thermal decoupling CVD method is used, comprising the following specific contents:
Step one, flowing a mixture gas of 100sccm He and 400sccm H2 in a tubular furnace for 10 minutes while heating the furnace to 775 ℃;
inserting the wafer into a furnace by using a magnetic coupling transfer arm, and then maintaining the same gas and flow rate as those in the first step, and maintaining the furnace at the temperature of 775 ℃ for 10 minutes;
changing the gas and the flow rate into a mixed gas of 100sccm of C2H4, 400sccm of He and 100sccm of H2 for 3min, and heating at 775 ℃; after growing carbon nanotubes on a wafer, keeping the same gas and flow, cooling the furnace to below 100 ℃, and finally purging with 1000sccm He for 5min to obtain a carbon capillary array;
step four, depositing a layer of SiO on the silicon substrate 2 In SiO 2 Spin-coating photoresist on the surface, etching part of the photoresist by using MEMS lithography technology to obtain a planar mask of the salt bridge (5) injection hole, covering part of the planar mask to obtain each injection hole, and etching SiO by HF 2 Depositing a layer, etching SiO not protected by photoresist 2 Part, obtain the three-dimensional structure of each injection hole, remove the photoresist mask, get the SiO of each injection hole 2 A hard mask;
step five, covering the SiO obtained in the step four on the carbon capillary array obtained in the step two 2 Hard mask, etching by KOH etching carbon capillary array with mass concentration of 35%, etching by SiO 2 The carbon capillary array part not protected by the hard mask obtains the three-dimensional structure of each injection hole and removes SiO 2 The hard mask, a salt bridge (5) with individual injection holes is obtained.
10. A method for preparing a sensitive film in a micro-integrated device for measuring electrolyte content of urine according to any one of claims 1 to 5, comprising the steps of:
firstly, preparing 20ml of selective membrane solution with selective responses to sodium, potassium and calcium ions respectively; wherein, the selective membrane solution of three ions uses one or two of cyclohexanone and tetrahydrofuran as mixed solvent; sodium tetraphenylborate and potassium tetraphenylborate as ion additives; polyvinyl chloride is used as a PVC matrix, and dioctyl sebacate is used as a plasticizer;
secondly, respectively dripping the prepared selective membrane solution on a wafer substrate, and spin-coating uniformly;
and thirdly, volatilizing the mixed solvent at room temperature to obtain a sensitive film, and cutting the sensitive film into a shape with proper size.
When the sodium ion selective membrane solution is prepared, tetraphenyl 24-crown ether-8 is selected as a sodium ion carrier, and the mass ratio of the ion additive, the ion carrier, the polyvinyl chloride and the dioctyl sebacate is 1.8:0.2:24:74;
When the potassium ion selective membrane solution is prepared, 1' -binaphthyl-20-crown-6 is selected as a carrier of potassium ions, and the mass ratio of an ion additive to the ion carrier to the polyvinyl chloride to the dioctyl sebacate is 1.8:0.2:33: 65.
When the calcium ion selective membrane solution is prepared, 1-tri (N-methyl-N-phenyl amino carbonyl methoxymethyl) propane is selected as a carrier of calcium ions, and an ionic additive, an ionic carrier, polyvinyl chloride and dioctyl sebacate are mixed according to the mass ratio of 1.8:0.2:32:66;
the mixed solvent is 20ml mixed solvent of cyclohexanone and tetrahydrofuran.
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