CN117420187A - Blood analysis sensor for monitoring dialysis progress and preparation method thereof - Google Patents

Blood analysis sensor for monitoring dialysis progress and preparation method thereof Download PDF

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CN117420187A
CN117420187A CN202311194979.6A CN202311194979A CN117420187A CN 117420187 A CN117420187 A CN 117420187A CN 202311194979 A CN202311194979 A CN 202311194979A CN 117420187 A CN117420187 A CN 117420187A
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electrode
antifouling
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adhesion
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孙雪梅
虞思汇
官航
彭慧胜
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Fudan University
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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
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Abstract

The invention belongs to the technical field of medical monitoring equipment, and particularly relates to a blood analysis sensor suitable for monitoring the dialysis progress of a patient and a preparation method thereof. The blood analysis sensor is based on a three-electrode system; the reference electrode is silver/silver chloride, and the surface of the reference electrode is not modified or modified with a bonding layer, an antifouling and anti-adhesion layer; the counter electrode is made of metal platinum, and the surface of the counter electrode is sequentially decorated with a bonding layer and an antifouling and anti-adhesion layer; the working electrode is made of metal platinum, and the surface of the working electrode is sequentially modified with a transduction layer, an ion selection layer, a bonding layer and an antifouling and anti-adhesion layer; the blood analysis sensor has good antifouling and anti-adhesion capabilities and can prevent biofouling; the method is used for monitoring the dialysis progress and comprises the steps of monitoring alternating current impedance parameters under characteristic frequency and estimating the water content of blood according to a fitting curve; monitoring changes in the concentration of the primary electrolyte in the blood; the combination of the two functions can help a physician determine the amount of ultrafiltration, the duration of dialysis and the ultrafiltration rate that the patient receives.

Description

Blood analysis sensor for monitoring dialysis progress and preparation method thereof
Technical Field
The invention belongs to the field of medical monitoring equipment, and particularly relates to a blood analysis sensor for monitoring the dialysis progress of a patient and a preparation method thereof.
Background
Regular hemodialysis treatment is an important means of maintaining survival of end stage renal patients. In dialysis treatment, the dialysis duration, ultrafiltration volume and ultrafiltration rate of a patient are reasonably adjusted to directly determine the treatment efficiency and safety.
In clinical practice, problems of cardiovascular load due to insufficient dialysis hypotension and ultrafiltration occur. At present, the regulation and control of the dialysis duration and rate is mainly based on the electrical impedance test of the human body, and the physician estimates the dry weight. The electrical impedance method which is developed in recent years can accurately reflect the hematocrit based on the capacitance of blood cell membranes and the resistance of cytoplasms, and provides a new thought for determining relative blood volume and analyzing blood composition. Meanwhile, since blood contains a large amount of protein, the direct contact of the metal electrode with the blood sample can lead to biomass accumulation on the surface of the electrode, and serious adverse effects are brought to the prediction of hematocrit. Therefore, developing an electrode that prevents biomass fouling and monitors electrolyte concentration during dialysis is a challenge.
Disclosure of Invention
The invention provides an anti-biofouling blood analysis sensor and a preparation method thereof, aiming at the problem that ultrafiltration volume and ultrafiltration speed are difficult to determine in dialysis treatment.
The invention monitors the hematocrit of the blood through the electrochemical impedance spectrum, calculates the water content of the blood, and monitors the main electrolyte concentration in the blood, thereby providing a basis for personalized regulation and control of the ultrafiltration speed and ultrafiltration volume of a patient and assisting in judging the dialysis end point.
The invention provides a blood analysis sensor for monitoring the dialysis progress of a patient, which is a three-electrode system, namely, the sensor consists of a reference electrode, a working electrode and a counter electrode; wherein:
the reference electrode comprises silver/silver chloride (namely a substrate electrode) as a main component; the surface of the material can be sequentially modified with an adhesive layer and an antifouling and anti-adhesive layer;
the main component of the counter electrode is metal platinum (namely a substrate electrode), and the surface of the counter electrode is sequentially modified with a bonding layer and an antifouling and anti-adhesion layer;
the main components of the working electrode are metal platinum (namely a substrate electrode), and the working electrode is divided into two types according to different analysis objects and different modification materials:
one is a sensor for hematocrit analysis, wherein a bonding layer and an anti-fouling and anti-adhesion layer are sequentially modified on the surface of a substrate electrode;
one is a sensor for analyzing electrolyte concentration, which is characterized in that a transduction layer, an ion selection layer, a bonding layer and an antifouling and anti-adhesion layer are sequentially modified on the surface of a substrate electrode;
the modified transduction layer is used for stabilizing the electron transmission process of the working electrode, increasing the redox capacitance and obtaining a more stable electric signal;
the ion selective layer material comprises: a polymer matrix, an ionophore, an ion exchanger and a plasticizer, and dispersed with a volatile solvent; wherein:
the polymer matrix provides mechanical support for the membrane layer and is an entrapping material; the ionophore is used for combining with target ions to provide selectivity for the sensor; the ion exchanger is used for providing the movement of the counter ion and the auxiliary ion carrier between different functional layers; the plasticizer is used for enhancing the movement capability of polymer molecular chains in a polymer matrix and improving the signal transmission speed;
the bonding layer is used for providing strong adhesion capability and bonding the antifouling anti-adhesion layer and the functional layer;
the antifouling and anti-adhesion layer is used for preventing adhesion of various biological pollutants such as proteins, cells and the like.
Further:
the transduction layer material is conductive high polymer material poly (3, 4-ethylenedioxythiophene) -poly (styrenesulfonic acid) polymer;
the ion selective layer material comprises: a polymer matrix, an ionophore, an ion exchanger and a plasticizer, and dispersed with a volatile solvent; wherein the polymer matrix is selected from polyvinyl chloride and styrene block copolymers; the ionophore is selected from valinomycin, 4-tert-butylcalixarene-ethyl tetraacetate, N, N, N ', N' -tetracyclohexyl-3-oxaglutaramide and the like; the ion exchanger is selected from sodium tetraphenyl borate, sodium tetrakis [3, 5-bis (trifluoromethyl) phenyl ] borate and the like; the plasticizer is selected from diisooctyl sebacate, o-nitrophenyl octyl ether and the like; the volatile solvent is selected from cyclohexanone and tetrahydrofuran.
The bonding layer material is small molecule dopamine, and has strong adhesive capacity;
the antifouling and anti-adhesion layer material is 2-methacryloyloxyethyl phosphorylcholine, and the antifouling and anti-adhesion effects on various biological pollutants such as proteins, cells and the like are realized by utilizing the zwitterionic charges of phosphorylcholine groups.
The invention also provides a preparation method of the blood analysis sensor, wherein:
preparation of working electrode:
the method is specifically divided into two types:
one is a sensor for hematocrit analysis, wherein a bonding layer and an anti-fouling and anti-adhesion layer are sequentially modified on the surface of a substrate electrode;
one is a sensor for analyzing electrolyte concentration, which is characterized in that a transduction layer, an ion selection layer, a bonding layer and an antifouling and anti-adhesion layer are sequentially modified on the surface of a substrate electrode; specifically:
the electrode modification steps for the sensor for electrolyte concentration analysis are:
(1) Modifying the transduction layer; preparing stable transduction layer dispersion liquid by using a conductive polymer material with high redox volume capacitance, uniformly coating the transduction layer dispersion liquid on the corresponding electrode position, and drying to obtain a working electrode modified with a transduction layer;
(2) Modifying the ion selection layer; preparing a polymer ion selective membrane solution by using a volatile organic solvent, wherein the polymer ion selective membrane solution comprises auxiliary materials such as a plasticizer, an ionophore, an ion exchanger and the like, so that the chain movement capability is enhanced, and the ion selectivity of a membrane layer is endowed; uniformly coating the ion selective membrane solution on the surface of the electrode modified with the transduction layer, and drying to obtain the electrode modified with the ion selective layer;
(3) Modifying the bonding layer; preparing a small molecular material solution with stable dispersion, soaking a conductive path in a bonding solution, adding a pH regulator to enable small molecules to self-polymerize, and forming a bonding layer on the conductive path;
(4) Modifying the antifouling and anti-sticking layer; preparing an anti-fouling and anti-adhesion material solution with stable dispersion, and soaking the conductive path modified with the bonding layer in the anti-fouling and anti-adhesion solution to enable the anti-fouling and anti-adhesion functional molecules to be modified on the electrode.
Electrode modification of the sensor for hematocrit analysis is performed by performing steps (3) and (4);
(II) preparation of a counter electrode: the surface of the electrode with the main component of metal platinum is sequentially modified with a bonding layer and an antifouling and anti-adhesion layer; the method is the same as that of the bonding layer, the antifouling and anti-sticking layer on the working electrode;
(III) preparation of a reference electrode: the surface of the electrode with the main component of silver/silver chloride is sequentially modified with a bonding layer and an antifouling and anti-adhesion layer; the method is the same as that of the bonding layer, the antifouling and anti-sticking layer on the working electrode; the surface of the reference electrode can be also not modified with a bonding layer and an antifouling and anti-adhesion layer.
Three base electrodes in the three-electrode system are constructed by a micro-nano processing method. Specifically, the shape of the electrode may be circular or rectangular, the size of the electrode is not more than 3mm by 3mm, and the distance between the electrode and the electrode is not more than 1mm. The electrodes are arranged in an array form, and the conductive paths are led out from the electrodes to the rear end interface according to the principle of mutual noninterruption.
Further:
in the modification of the transduction layer, the conductive polymer is poly (3, 4-ethylenedioxythiophene) -poly (styrenesulfonic acid) polymer; the solvent for dissolving the conductive polymer is water, the mass fraction is 80-95%, and the coating amount is 1-10 mu l.
In the ion selective layer modification, the polymer matrix is selected from polyvinyl chloride and styrene block copolymers; the ionophore is selected from valinomycin, 4-tert-butylcalixarene-ethyl tetraacetate, N, N, N ', N' -tetracyclohexyl-3-oxaglutaramide and the like; the ion exchanger is selected from sodium tetraphenyl borate, sodium tetrakis [3, 5-bis (trifluoromethyl) phenyl ] borate and the like; the plasticizer is selected from diisooctyl sebacate, o-nitrophenyl octyl ether and the like; the volatile solvent is cyclohexanone or tetrahydrofuran, etc.; the coating amount is 1. Mu.l to 5. Mu.l.
In the modification of the bonding layer, the cohesive small molecule is dopamine, the concentration is 0.1% -1.0%, the pH value is 7.4-9.0, and the polymerization time is 0.5-5 hours.
In the modification of the anti-fouling and anti-adhesion layer, the anti-fouling and anti-adhesion functional molecule is 2-methacryloyloxyethyl phosphorylcholine, the concentration is 0.2-4.0%, and the soaking time is 2-36 hours.
The blood analysis sensor is used for monitoring the dialysis progress, and the specific method is as follows:
(1) Monitoring alternating current impedance parameters under characteristic frequency, and estimating the water content of blood according to a fitting curve
(2) Monitoring the change in concentration of the primary electrolyte in the blood based on the reading of the ion-selective electrode;
the characteristic frequency can be 100Hz, 500Hz, 1000Hz and 5000Hz, and the measured alternating current impedance parameters are impedance, real part impedance, imaginary part impedance and phase angle.
A functional curve of impedance versus hematocrit can be fitted to the electrochemical impedance parameters to help a physician determine the duration and rate of dialysis treatment that a patient is receiving.
During measurement, the number of the electrodes can be flexibly adjusted according to the needs.
The blood analysis sensor can assist doctors to determine ultrafiltration volume and ultrafiltration speed of dialysis patients.
The invention has the beneficial effects that:
(1) One or more parameters of the hematocrit, the potassium ion concentration, the calcium ion concentration, the sodium ion concentration, the hydrogen ion concentration and the like can be detected simultaneously;
(2) The blood analysis sensor prepared by the invention modifies the antifouling anti-adhesion film, has good antifouling anti-adhesion capability, and can prevent biofouling on the surface of an electrode to cause measurement errors;
(3) Under the complex sample environments such as human blood, the performance of the sensor is stable after continuous testing for 5 hours; the detection effect on the hematocrit is equal to that of the main stream instrument on the market.
Drawings
FIG. 1 is a schematic diagram showing the overall structure of a blood analysis sensor.
Fig. 2 is a schematic diagram of the basic form of the sensor electrode portion (4 electrodes).
Fig. 3 shows the results of the sensor of example 1 on blood with different water contents and the fitting results.
FIG. 4 shows the results of the sensor of example 2 for blood with different sodium ion concentrations and the fitting results.
FIG. 5 shows the results of the sensor of example 3 for blood with different potassium ion concentrations and the fitting results.
Fig. 6 is a comparative example: results of the test for potassium ion concentration in blood with the anti-fouling and anti-adhesion layer unmodified. Wherein, (a) is the original data obtained by the electrode, the potential is unstable and gradually decreases along with time, and (b) is the relation between the concentration of potassium ions and the potential, the related data and the Nernst equation have deviation, and the fitting condition is poor.
FIG. 7 is a graph showing the results of testing the concentration of potassium ions in blood by the sensor of example 3 in the case of modifying the antifouling anti-adhesion layer. The method comprises the steps of (c) obtaining original data of the electrode, wherein the potential is stable and is in a ladder shape, and (d) obtaining the relation between the concentration of potassium ions and the potential, wherein the related data accords with a Nernst equation, and the fitting condition is good.
Detailed Description
The invention is further described in the following specific examples with reference to the drawings.
Example 1, hematocrit sensor:
(1) Preparing a patterned electrode path by a micro-nano processing technology; patterning was achieved by micro-nano processing techniques to deposit 100nm of platinum on polyethylene terephthalate plastic sheets to prepare patterned electrode pathways.
(2) Preparing a reference electrode;
(1) 395.5mg of polyvinyl butyral is dissolved in 5ml of methanol, and 250mg of sodium chloride and 250mg of silver nitrate are added after ultrasonic dispersion; shaking for 30min in dark, and reacting to obtain silver/silver chloride; dripping 5 mu l of the solution on the surface of the electrode path, and drying to obtain a reference electrode;
(2) the anti-fouling and anti-adhesion layer is modified on the reference electrode, and the specific steps are as follows:
first, a tie layer was prepared on a reference electrode: dissolving 2mg of dopamine in 10ml of 1mol/L tris hydrochloride buffer solution with pH value of 8.0, soaking a sensor electrode in the solution for 2 hours, and taking out to obtain a reference electrode with a bonding layer;
then, an antifouling and anti-adhesion layer is modified on the adhesive layer: 10mg of 2-methacryloyloxyethyl phosphorylcholine is taken and dissolved in 10ml of 1mol/L tris hydrochloride buffer solution with pH=8.0, a sensor electrode is soaked in the solution for 24 hours, and the reference electrode with an antifouling and anti-adhesion layer is obtained after the sensor electrode is taken out.
(3) Preparing a counter electrode;
(1) and (3) modifying an adhesive layer on the platinum electrode: dissolving 2mg of dopamine in 10ml of 1mol/L tris hydrochloride buffer solution with pH value of 8.0, soaking a sensor electrode in the solution for 2 hours, and taking out to obtain a counter electrode with a bonding layer;
(2) an antifouling and anti-sticking layer is modified on a platinum electrode with a bonding layer: 10mg of 2-methacryloyloxyethyl phosphorylcholine is taken and dissolved in 10ml of 1mol/L tris hydrochloride buffer solution with pH=8.0, a sensor electrode is soaked in the solution for 24 hours, and the counter electrode with an antifouling and anti-adhesion layer is obtained after the sensor electrode is taken out.
(4) Preparing a hematocrit working electrode;
(1) and (3) modifying an adhesive layer on the platinum electrode: dissolving 2mg of dopamine in 10ml of 1mol/L tris hydrochloride buffer solution with pH value of 8.0, soaking a sensor electrode in the solution for 2 hours, and taking out to obtain a working electrode with a bonding layer;
(2) an antifouling and anti-sticking layer is modified on a platinum electrode with a bonding layer: 10mg of 2-methacryloyloxyethyl phosphorylcholine is taken and dissolved in 10ml of 1mol/L tris hydrochloride buffer solution with pH=8.0, a sensor electrode is soaked in the solution for 24 hours, and the working electrode with an antifouling and anti-adhesion layer is obtained after the sensor electrode is taken out.
(5) Performing a red blood cell packed volume measurement; and the electrochemical workstation is connected with the reference electrode, the counter electrode and the working electrode which are prepared according to the method, and the alternating current impedance parameters under the frequencies of 100Hz, 1000Hz and 10000Hz are measured.
(6) And drawing an impedance-red blood cell pressure product function curve according to the alternating current impedance test result.
The results of testing blood with different water contents and fitting curves are shown in fig. 3.
Example 2, blood moisture-electrolyte analysis sensor in dialysis:
(1) Preparing a patterned electrode path by a micro-nano processing technology; patterning was achieved by micro-nano processing techniques to deposit 100nm of platinum on polyethylene terephthalate plastic sheets to prepare patterned electrode pathways.
(2) Preparing a reference electrode;
395.5mg of polyvinyl butyral is dissolved in 5ml of methanol, and 250mg of sodium chloride and 250mg of silver nitrate are added after ultrasonic dispersion; shaking for 30min in dark; and 5. Mu.l of the solution is dripped on the surface of the electrode path, and the reference electrode is obtained after drying.
(3) Preparing a counter electrode;
(1) and (3) modifying an adhesive layer on the platinum electrode: dissolving 2mg of dopamine in 10ml of 1mol/L tris hydrochloride buffer solution with pH value of 8.0, soaking a sensor electrode in the solution for 2 hours, and taking out to obtain a counter electrode with a bonding layer;
(2) an antifouling and anti-sticking layer is modified on a platinum electrode with a bonding layer: 10mg of 2-methacryloyloxyethyl phosphorylcholine is taken and dissolved in 10ml of 1mol/L tris hydrochloride buffer solution with pH=8.0, a sensor electrode is soaked in the solution for 24 hours, and the counter electrode with an antifouling and anti-adhesion layer is obtained after the sensor electrode is taken out.
(4) Preparing a hematocrit working electrode;
(1) and (3) modifying an adhesive layer on the platinum electrode: dissolving 2mg of dopamine in 10ml of 1mol/L tris hydrochloride buffer solution with pH value of 8.0, soaking a sensor electrode in the solution for 2 hours, and taking out to obtain a working electrode with a bonding layer;
(2) an antifouling and anti-sticking layer is modified on a platinum electrode with a bonding layer: 10mg of 2-methacryloyloxyethyl phosphorylcholine is taken and dissolved in 10ml of 1mol/L tris hydrochloride buffer solution with pH=8.0, a sensor electrode is soaked in the solution for 24 hours, and the working electrode with an antifouling and anti-adhesion layer is obtained after the sensor electrode is taken out.
(5) Preparing an electrolyte concentration sensing working electrode;
(1) preparing a transduction layer on the surface of a platinum electrode; 2ml of poly-3, 4-ethylenedioxythiophene pre-doped with sodium polystyrene sulfonate was mixed with 10. Mu.l of dodecylbenzenesulfonic acid, 100. Mu.l of ethylene glycol and stirred for 3 hours; adding 9.6 mu l of cross-linking agent 3- (2, 3-glycidoxy) propyl trimethoxy silane, and stirring for 4 hours to obtain a transduction layer solution; 2.5 mu l of the solution is dripped on the surface of the electrode, thus obtaining a platinum electrode with a transduction layer;
(2) preparing an ion selection layer on the surface of the transduction layer; 10mg of valinomycin, 2.5mg of potassium tetra (4-chlorophenyl) borate, 163.5mg of high molecular weight polyvinyl chloride and 323.5mg of diisooctyl sebacate are mixed, 1750mg of cyclohexanone is added for dissolution, and stirring is carried out overnight, thus obtaining a potassium ion selective membrane solution; coating 3 μl of the solution on the corresponding position of the electrode, and drying to obtain ion-selective electrode;
(3) modifying an antifouling and anti-adhesion layer on the surface of the ion selection layer;
first, a bonding layer is modified on an ion selective layer: dissolving 2mg of dopamine in 10ml of 1mol/L tris hydrochloride buffer solution with pH value of 8.0, soaking a sensor electrode in the solution for 2 hours, and taking out to obtain an ion selective electrode with a bonding layer;
then, an antifouling and anti-sticking layer is modified on the ion-selective electrode with the bonding layer: 10mg of 2-methacryloyloxyethyl phosphorylcholine is taken and dissolved in 10ml of 1mol/L tris hydrochloride buffer solution with pH=8.0, a sensor electrode is soaked in the solution for 24 hours, and the ion-selective electrode with an antifouling and anti-adhesion layer is obtained after the sensor electrode is taken out.
(5) The hematocrit measurement was performed. The electrochemical workstation is connected, the reference electrode, the counter electrode and the red blood cell pressure product working electrode prepared by the method are connected according to a three-electrode system, and the alternating current impedance parameters under the frequencies of 100Hz, 1000Hz and 10000Hz are measured.
(6) And drawing an impedance-red blood cell pressure product function curve according to the alternating current impedance test result, and calculating the moisture content in blood.
(7) Electrolyte concentration measurements were made. And the electrochemical working stations are connected, the electrolyte concentration sensing working electrodes prepared according to the method are respectively connected, the open-circuit potential difference is measured, the electrolyte concentration is read, and the electrolyte concentration is monitored in real time.
The results of the detection and fitting of blood with different potassium ion concentrations are shown in fig. 4.
Example 3, multifunctional blood analysis sensor in dialysis:
(1) Patterned electrode vias were prepared by micro-nano processing techniques. The patterning deposition of 80nm platinum on polyethylene terephthalate plastic sheet is realized by micro-nano processing technology, and the patterned electrode path is prepared.
(2) Preparation of reference electrode
And 3 mu l of silver/silver chloride slurry is dripped on the surface of the electrode path, the electrode path is dried for 15 hours in a dark environment, 3 mu l of 7% polyvinyl butyral methanol solution is dripped, and the reference electrode is obtained after drying.
(3) Preparation of counter electrode
(1) And (3) modifying an adhesive layer on the platinum electrode: dissolving 4mg of dopamine in 10ml of 0.5mol/L tris hydrochloride buffer solution with pH value of 8.0, soaking a sensor electrode in the solution for 2 hours, and taking out to obtain a counter electrode with a bonding layer;
(2) an antifouling and anti-sticking layer is modified on a platinum electrode with a bonding layer: 20mg of 2-methacryloyloxyethyl phosphorylcholine is taken and dissolved in 10ml of 0.5mol/L tris hydrochloride buffer solution with pH=8.0, a sensor electrode is soaked in the solution for 24 hours, and the counter electrode with an antifouling and anti-adhesion layer is obtained after the sensor electrode is taken out.
(4) Preparation of a hematocrit working electrode
(1) And (3) modifying an adhesive layer on the platinum electrode: dissolving 4mg of dopamine in 10ml of 0.5mol/L tris hydrochloride buffer solution with pH value of 8.0, soaking a sensor electrode in the solution for 2 hours, and taking out to obtain a working electrode with a bonding layer;
(2) an antifouling and anti-sticking layer is modified on a platinum electrode with a bonding layer: 20mg of 2-methacryloyloxyethyl phosphorylcholine is taken and dissolved in 10ml of 0.5mol/L tris hydrochloride buffer solution with pH=8.0, a sensor electrode is soaked in the solution for 24 hours, and the working electrode with an antifouling and anti-adhesion layer is obtained after the sensor electrode is taken out.
(5) Preparation of electrolyte concentration sensing working electrode
(1) Preparing a transduction layer on the surface of a platinum electrode; 2ml of poly-3, 4-ethylenedioxythiophene pre-doped with sodium polystyrene sulfonate was mixed with 10. Mu.l of dodecylbenzenesulfonic acid, 150. Mu.l of ethylene glycol and stirred for 2 hours; adding 9.6 mu l of cross-linking agent 3- (2, 3-glycidoxy) propyl trimethoxy silane, and stirring for 4 hours to obtain a transduction layer solution; 2.5 mu l of the solution is dripped on the surface of the electrode, thus obtaining a platinum electrode with a transduction layer;
(2) preparing an ion selection layer on the surface of the transduction layer; 5mg of 4-tert-butylcalix [4] arene-ethyl tetraacetate, 2.75mg of sodium tetrakis [3, 5-bis (trifluoromethyl) phenyl ] borate, 210mg of styrene block copolymer, 327.25mg of 2-nitrophenyl n-octyl ether and 3300 mu l of tetrahydrofuran are mixed and stirred overnight, so that a sodium ion selective membrane solution is obtained; 10mg of valinomycin, 2.5mg of potassium tetra (4-chlorophenyl) borate, 163.5mg of high molecular weight polyvinyl chloride and 323.5mg of diisooctyl sebacate are mixed, 1750mg of cyclohexanone is added for dissolution, and stirring is carried out overnight, thus obtaining a potassium ion selective membrane solution; coating 3 μl of the above solutions on the corresponding positions of the electrodes, and drying to obtain ion-selective electrodes; (other electrolyte sensing components can be flexibly selected as required);
(3) modifying the surface of ion selective layer with antifouling and anti-sticking layer
First, a bonding layer is modified on an ion selective layer: dissolving 4mg of dopamine in 10ml of 0.5mol/L tris hydrochloride buffer solution with pH value of 8.0, soaking a sensor electrode in the solution for 2 hours, and taking out to obtain an ion selective electrode with a bonding layer;
then, an antifouling and anti-sticking layer is modified on the ion-selective electrode with the bonding layer: 20mg of 2-methacryloyloxyethyl phosphorylcholine is taken and dissolved in 10ml of 0.5mol/L tris hydrochloride buffer solution with pH=8.0, a sensor electrode is soaked in the solution for 24 hours, and the ion-selective electrode with an antifouling and anti-adhesion layer is obtained after the sensor electrode is taken out.
(5) Performing a red blood cell packed volume measurement; the electrochemical workstation is connected, the reference electrode, the counter electrode and the red blood cell pressure product working electrode prepared by the method are connected according to a three-electrode system, and the alternating current impedance parameters under the frequencies of 100Hz, 1000Hz and 10000Hz are measured.
(6) And drawing an impedance-red blood cell pressure product function curve according to the alternating current impedance test result, and calculating the moisture content in blood.
(7) Electrolyte concentration measurements were made. And the electrochemical working stations are connected, the electrolyte concentration sensing working electrodes prepared according to the method are respectively connected, the open-circuit potential difference is measured, the electrolyte concentration is read, and the electrolyte concentration is monitored in real time.
(8) Based on the measurement results, the ultrafiltration volume and ultrafiltration rate of the patient are reasonably arranged.
The results of the detection and fitting of blood with different sodium ion concentrations are shown in fig. 5.
The results of the test of the sensor of example 3 for the concentration of potassium ions in blood with the stain-resistant and anti-adhesion layer modified are shown in FIG. 7.

Claims (8)

1. A blood analysis sensor for monitoring the dialysis progress of a patient, which is characterized by a three-electrode system, namely, a reference electrode, a working electrode and a counter electrode; wherein:
the reference electrode comprises silver/silver chloride as a main component; the surface of the adhesive is not modified or is sequentially modified with an adhesive layer, an antifouling and anti-adhesive layer;
the main component of the counter electrode is metal platinum, and the surface of the counter electrode is sequentially modified with a bonding layer and an antifouling and anti-adhesion layer;
the main component of the working electrode is metal platinum, and according to different analysis objects, the modification materials are different, and the working electrode is specifically divided into two types:
one is a sensor for hematocrit analysis, wherein a bonding layer and an anti-fouling and anti-adhesion layer are sequentially modified on the surface of a substrate electrode;
one is a sensor for analyzing electrolyte concentration, which is characterized in that a transduction layer, an ion selection layer, a bonding layer and an antifouling and anti-adhesion layer are sequentially modified on the surface of a substrate electrode;
the modified transduction layer is used for stabilizing the electron transmission process of the working electrode, increasing the redox capacitance and obtaining a more stable electric signal;
the ion selective layer material comprises: a polymer matrix, an ionophore, an ion exchanger and a plasticizer, wherein:
the polymer matrix provides mechanical support for the membrane layer and is an entrapping material; the ionophore is used for combining with target ions to provide selectivity for the sensor; the ion exchanger is used for providing the movement of the counter ion and the auxiliary ion carrier between different functional layers; the plasticizer is used for enhancing the movement capability of polymer molecular chains in a polymer matrix and improving the signal transmission speed;
the bonding layer is used for providing strong adhesion capability and bonding the antifouling anti-adhesion layer and the functional layer;
the antifouling and anti-adhesion layer is used for preventing adhesion of biological pollutants.
2. The blood analysis sensor of claim 1, wherein:
the transduction layer material is conductive high polymer material poly (3, 4-ethylenedioxythiophene) -poly (sodium styrenesulfonate) polymer;
in the ion selective layer material, the ionophore is selected from valinomycin, 4-tert-butylcalixarene-ethyl tetraacetate and N, N, N ', N' -tetracyclohexyl-3-oxaglutaramide; the polymer matrix is polyvinyl chloride or styrene block copolymer; the plasticizer is diisooctyl sebacate or o-nitrophenyl octyl ether;
the bonding layer material is small molecule dopamine;
the antifouling and anti-adhesion layer material is 2-methacryloyloxyethyl phosphorylcholine.
3. A method of manufacturing a blood analysis sensor according to claim 1 or 2, comprising:
the preparation of the working electrode is divided into two types:
one is a sensor for hematocrit analysis, wherein a bonding layer and an anti-fouling and anti-adhesion layer are sequentially modified on the surface of a substrate electrode;
one is a sensor for analyzing electrolyte concentration, which is characterized in that a transduction layer, an ion selection layer, a bonding layer and an antifouling and anti-adhesion layer are sequentially modified on the surface of a substrate electrode; specifically:
the electrode modification steps for the sensor for electrolyte concentration analysis are:
(1) Modifying the transduction layer; preparing stable transduction layer dispersion liquid by using a conductive polymer material with high redox volume capacitance, uniformly coating the transduction layer dispersion liquid on the corresponding electrode position, and drying to obtain a working electrode modified with a transduction layer;
(2) Modifying the ion selection layer; preparing a polymer ion selective membrane solution by using a volatile organic solvent, wherein the polymer ion selective membrane solution comprises a plasticizer, an ionophore and an ion exchanger auxiliary material, so that the chain movement capacity is enhanced, and the membrane layer ion selectivity and the charge movement capacity are endowed; uniformly coating the ion selective membrane solution on the surface of the electrode modified with the transduction layer, and drying to obtain the electrode modified with the ion selective layer;
(3) Modifying the bonding layer; preparing a bonding micromolecule solution with stable dispersion, soaking a conductive path in the bonding solution, adding a pH regulator to enable micromolecules to self-polymerize, and forming a bonding layer on the conductive path;
(4) Modifying the antifouling and anti-sticking layer; preparing an antifouling and anti-adhesion material solution with stable dispersion, and soaking a conductive path modified with a bonding layer in the antifouling and anti-adhesion solution to enable antifouling and anti-adhesion functional molecules to be modified on an electrode;
the electrode modification of the sensor for hematocrit analysis is performed by steps (3) and (4);
(II) preparation of a counter electrode: the surface of the electrode with the main component of metal platinum is sequentially modified with a bonding layer and an antifouling and anti-adhesion layer; the method is characterized in that a bonding layer and an antifouling and anti-sticking layer are modified on a working electrode;
(III) preparation of a reference electrode: sequentially modifying a bonding layer and an antifouling and anti-adhesion layer on an electrode with silver/silver chloride as a main component; the method is characterized in that a bonding layer and an antifouling and anti-sticking layer are modified on a working electrode; or the reference electrode does not modify the bonding layer, the antifouling and anti-adhesion layer;
three base electrodes in the three-electrode system are constructed by a micro-nano processing method.
4. A method of preparation according to claim 3, characterized in that:
in the modification of the transduction layer, the conductive polymer is poly (3, 4-ethylenedioxythiophene) -poly (styrenesulfonic acid) polymer; the solvent for dissolving the conductive polymer is water, and the mass fraction is 80-95%;
in the ion selective layer modification, the polymer matrix is selected from polyvinyl chloride and styrene block copolymers; the ionophore is selected from valinomycin, 4-tert-butylcalixarene-ethyl tetraacetate, N, N, N ', N' -tetracyclohexyl-3-oxaglutaramide and the like; the ion exchanger is selected from sodium tetraphenyl borate and sodium tetrakis [3, 5-bis (trifluoromethyl) phenyl ] borate; the plasticizer is selected from diisooctyl sebacate and o-nitrophenyl octyl ether; the volatile solvent is selected from cyclohexanone and tetrahydrofuran;
in the modification of the bonding layer, the cohesive small molecules are dopamine, the concentration is 0.1% -1.0%, the pH value is 7.4-9.0, and the polymerization time is 0.5-5 hours;
in the modification of the anti-fouling and anti-adhesion layer, the anti-fouling and anti-adhesion functional molecule is 2-methacryloyloxyethyl phosphorylcholine, the concentration is 0.2-4.0%, and the soaking time is 2-36 hours.
5. Use of a blood analysis sensor according to claim 1 or 2 for monitoring the progress of dialysis, in particular by:
(1) Monitoring alternating current impedance parameters under characteristic frequency, and estimating the water content of blood according to a fitting curve;
(2) Based on the reading of the ion selective electrode, the change in concentration of the primary electrolyte in the blood is monitored.
6. The use according to claim 5, wherein the characteristic frequencies are 100Hz, 500Hz, 1000Hz, 5000Hz and the measured characteristic electrical impedance parameters are impedance, real impedance, imaginary impedance and phase angle.
7. The use of claim 6, wherein the impedance-hematocrit function is fitted based on the electrochemical impedance parameters to assist the physician in determining the duration of the patient undergoing dialysis treatment and the dialysis rate.
8. Use according to claims 4-7, characterized in that the number of electrodes is flexibly adjusted according to the detection need.
CN202311194979.6A 2023-09-15 2023-09-15 Blood analysis sensor for monitoring dialysis progress and preparation method thereof Pending CN117420187A (en)

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