CN114487063A - Flexible sweat lactic acid electrochemical sensor with pH correction function - Google Patents
Flexible sweat lactic acid electrochemical sensor with pH correction function Download PDFInfo
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- CN114487063A CN114487063A CN202210096856.8A CN202210096856A CN114487063A CN 114487063 A CN114487063 A CN 114487063A CN 202210096856 A CN202210096856 A CN 202210096856A CN 114487063 A CN114487063 A CN 114487063A
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- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 title claims abstract description 96
- 210000004243 sweat Anatomy 0.000 title claims abstract description 51
- 239000004310 lactic acid Substances 0.000 title claims abstract description 48
- 235000014655 lactic acid Nutrition 0.000 title claims abstract description 48
- 238000012937 correction Methods 0.000 title claims abstract description 21
- JVTAAEKCZFNVCJ-UHFFFAOYSA-M Lactate Chemical compound CC(O)C([O-])=O JVTAAEKCZFNVCJ-UHFFFAOYSA-M 0.000 claims abstract description 27
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims abstract description 13
- 229910052737 gold Inorganic materials 0.000 claims abstract description 13
- 239000010931 gold Substances 0.000 claims abstract description 13
- 239000012528 membrane Substances 0.000 claims abstract description 10
- 239000000758 substrate Substances 0.000 claims abstract description 9
- 102000003855 L-lactate dehydrogenase Human genes 0.000 claims abstract description 8
- 108700023483 L-lactate dehydrogenases Proteins 0.000 claims abstract description 8
- 230000009144 enzymatic modification Effects 0.000 claims abstract description 7
- 239000007788 liquid Substances 0.000 claims abstract description 7
- 229920001661 Chitosan Polymers 0.000 claims description 5
- 238000000034 method Methods 0.000 claims description 4
- 238000012544 monitoring process Methods 0.000 claims description 4
- 239000002048 multi walled nanotube Substances 0.000 claims description 4
- 239000012488 sample solution Substances 0.000 claims description 4
- 229910021607 Silver chloride Inorganic materials 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims description 3
- HKZLPVFGJNLROG-UHFFFAOYSA-M silver monochloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 claims description 3
- 239000012530 fluid Substances 0.000 claims 1
- 238000001514 detection method Methods 0.000 abstract description 12
- 238000002360 preparation method Methods 0.000 abstract description 9
- 238000011161 development Methods 0.000 abstract description 3
- 230000005540 biological transmission Effects 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 15
- 210000004369 blood Anatomy 0.000 description 5
- 239000008280 blood Substances 0.000 description 5
- 239000000523 sample Substances 0.000 description 5
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 4
- 229910052709 silver Inorganic materials 0.000 description 4
- 239000004332 silver Substances 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 108010073450 Lactate 2-monooxygenase Proteins 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000010241 blood sampling Methods 0.000 description 2
- JHIVVAPYMSGYDF-UHFFFAOYSA-N cyclohexanone Chemical compound O=C1CCCCC1 JHIVVAPYMSGYDF-UHFFFAOYSA-N 0.000 description 2
- 230000014509 gene expression Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 206010016855 Foetal distress syndrome Diseases 0.000 description 1
- 229910021578 Iron(III) chloride Inorganic materials 0.000 description 1
- 230000006538 anaerobic glycolysis Effects 0.000 description 1
- 230000000386 athletic effect Effects 0.000 description 1
- 238000010923 batch production Methods 0.000 description 1
- 229910021538 borax Inorganic materials 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000035606 childbirth Effects 0.000 description 1
- 238000000970 chrono-amperometry Methods 0.000 description 1
- 239000005515 coenzyme Substances 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- VJHINFRRDQUWOJ-UHFFFAOYSA-N dioctyl sebacate Chemical compound CCCCC(CC)COC(=O)CCCCCCCCC(=O)OCC(CC)CCCC VJHINFRRDQUWOJ-UHFFFAOYSA-N 0.000 description 1
- 238000000835 electrochemical detection Methods 0.000 description 1
- 238000005566 electron beam evaporation Methods 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 229920002457 flexible plastic Polymers 0.000 description 1
- 210000001061 forehead Anatomy 0.000 description 1
- 230000010224 hepatic metabolism Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 208000006443 lactic acidosis Diseases 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000000663 muscle cell Anatomy 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 239000002985 plastic film Substances 0.000 description 1
- 229920000915 polyvinyl chloride Polymers 0.000 description 1
- 239000004800 polyvinyl chloride Substances 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000027756 respiratory electron transport chain Effects 0.000 description 1
- 210000003296 saliva Anatomy 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 235000010339 sodium tetraborate Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 210000001138 tear Anatomy 0.000 description 1
- SWZDQOUHBYYPJD-UHFFFAOYSA-N tridodecylamine Chemical compound CCCCCCCCCCCCN(CCCCCCCCCCCC)CCCCCCCCCCCC SWZDQOUHBYYPJD-UHFFFAOYSA-N 0.000 description 1
- 125000002023 trifluoromethyl group Chemical group FC(F)(F)* 0.000 description 1
- BSVBQGMMJUBVOD-UHFFFAOYSA-N trisodium borate Chemical compound [Na+].[Na+].[Na+].[O-]B([O-])[O-] BSVBQGMMJUBVOD-UHFFFAOYSA-N 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3271—Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
- G01N27/3272—Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3271—Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
- G01N27/3274—Corrective measures, e.g. error detection, compensation for temperature or hematocrit, calibration
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Electrochemistry (AREA)
- Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
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Abstract
The invention provides a flexible sweat lactic acid electrochemical sensor with pH correction, which comprises a flexible substrate, and a lactic acid sensing electrode, a pH electrode, a reference electrode and a counter electrode which are arranged on the surface of the flexible substrate. The lactic acid sensing electrode and the pH electrode are prepared by respectively dripping enzyme modification liquid and H + selective membrane solution on the surfaces of two independent flexible gold electrodes. The lactate sensing electrode utilizes lactate dehydrogenase to oxidize lactate in sweat and carries out electronic transmission with the electrode to generate a current signal, the pH electrode simultaneously measures the pH of a sweat sample, and finally, the lactate in the sweat is quantitatively analyzed based on a standard curve after pH correction. The flexible sweat lactic acid electrochemical sensor with pH correction provided by the invention has the advantages of simple preparation, good stability, low cost, high detection accuracy and the like, and provides good technical support for development of wearable sweat biosensing devices.
Description
One, the technical field
The invention relates to the technical field of electrochemical sensors, in particular to a flexible sweat lactic acid electrochemical sensor with pH correction.
Second, background Art
Lactic acid is a product of anaerobic glycolysis, blood lactic acid is mainly produced by muscle cells and erythrocytes via liver metabolism, and the blood lactic acid level reflects the comprehensive condition of production and metabolism of body substances, and has diagnostic significance for lactic acidosis when the blood lactic acid value of the body is more than 4 mmol/L. Moreover, the blood lactic acid measurement has important significance for evaluating the athletic competitive ability of athletes and truly and accurately reflecting the intrauterine hypoxia condition of the pregnant women and the childbirth period. The existing lactic acid detection method mainly comprises venous or arterial blood sampling detection. However, the high-frequency blood sampling not only brings great pressure to the body and the mind of a patient, but also has the risk of invasion and infection and cannot realize real-time monitoring.
The research proves that the concentration of lactic acid in biological liquid (such as sweat, tears, saliva and urine) secreted by human body has good positive correlation with the content level of lactic acid in blood. With the advent of the electronic information age, wearable non-invasive sweat lactate detection devices were developed and used for real-time monitoring of lactate. The wearable sweat lactate detection equipment based on lactate oxidase development has high detection sensitivity, but lactate oxidase is expensive and difficult to store, and is not beneficial to the production of non-invasive sweat lactate sensors. In order to reduce the preparation cost of the sensor, a wearable sweat lactate sensor based on lactate dehydrogenase is proposed, but coenzyme NAD is still required to be added+And the lactic acid detection result is interfered by the acidity of sweat, and the accuracy needs to be improved. Therefore, the development of a sweat lactate sensor which is simple in preparation, good in stability, low in cost and high in detection accuracy is urgently needed.
Third, the invention
The invention aims to provide a flexible sweat lactic acid electrochemical sensor which is high in detection accuracy, low in cost, simple to prepare and good in stability, and is used for developing a wearable sweat biosensor device.
The purpose of the invention can be realized by the following technical scheme:
a flexible sweat lactate electrochemical sensor with pH correction comprises a flexible substrate, and a lactate sensing electrode, a pH electrode, a reference electrode and a counter electrode which are arranged on the surface of the flexible substrate (figure 1). The lactic acid sensing electrode and the pH electrode are respectively prepared on the surfaces of two independent flexible gold electrodes; the lactate sensing electrode is prepared by directly dripping enzyme modification liquid containing multi-walled carbon nanotubes and lactate dehydrogenase on the surface of one flexible gold electrode; the pH electrode is formed by directly dripping H on the surface of another flexible gold electrode+And preparing a selective membrane solution. When sweat sample is covered on the sensor surface, lactate dehydrogenase on the lactate sensing electrode oxidizes lactate in sweat and carries out electron transfer with the electrode to generate current signal, and pH electrode is provided with a pH electrodeH of (A) to (B)+Selective membranes specifically respond only to H in sweat+And further monitoring the pH of the sweat in real time, and finally carrying out quantitative analysis on the lactic acid in the sweat based on the standard curve after pH correction.
The enzyme modification liquid is prepared by uniformly mixing multi-wall carbon nano tubes and lactate dehydrogenase in a certain proportion in a chitosan solution.
The reference electrode is an Ag/AgCl reference electrode, and the counter electrode is a gold electrode and is shared by the lactic acid sensing electrode and the pH electrode.
The flexible sweat lactic acid electrochemical sensor with the pH correction function can be attached to the parts, such as arms, forehead and the like, which are easy to sweat, so that the lactic acid in the sweat can be detected.
Compared with the prior art, the invention has the following beneficial effects:
(1) the lactate sensing electrode and the pH electrode on the sensor are formed by one-step dripping of enzyme modification liquid and H+The preparation of the selective membrane solution has simple and quick process and is suitable for batch production.
(2) The invention provides a method for quantitatively analyzing lactic acid in sweat based on a standard curve after pH correction, which effectively ensures the accuracy of the detection result of lactic acid in sweat.
(3) The sensor provided by the invention has the detection range of 0-40mM for lactic acid, covers the physiological concentration of human lactic acid, and has good clinical application.
(4) The flexible sweat lactic acid electrochemical sensor with pH correction provided by the invention can be flexibly integrated on flexible plastic films, fabrics and the like, and is used for developing wearable sweat biosensing devices.
Description of the drawings
FIG. 1 is a schematic diagram of the construction of a flexible sweat lactate electrochemical sensor with pH correction according to the present invention;
FIG. 2 is a standard curve of lactate detection in sample solutions of different pH for the flexible sweat lactate electrochemical sensor with pH correction according to the present invention.
Fifth, detailed description of the invention
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are exemplary of the invention, are intended to be illustrative of the invention, and are not intended to limit the invention.
The invented flexible sweat lactic acid electrochemical sensor with pH correction comprises a flexible substrate, and a lactic acid sensing electrode, a pH electrode, a reference electrode and a counter electrode which are arranged on the surface of the flexible substrate (figure 1).
The preparation process of the flexible sweat lactic acid electrochemical sensor with pH correction comprises the steps of preparing a flexible electrode, modifying enzyme solution and H+Preparation of selective membrane and its modification.
The flexible electrode is prepared by electron beam evaporation: on PET with the thickness of 100 mu m, different metal mask plates are used for sequentially evaporating gold and silver on specific positions to prepare a flexible electrode containing three gold electrodes and one silver electrode; one gold electrode is used for preparing the lactic acid sensing electrode, one gold electrode is used for preparing the pH electrode, the remaining gold electrode is used as the counter electrode, the silver electrode is used for preparing the Ag/AgCl reference electrode, and the counter electrode and the reference electrode are shared by the lactic acid sensing electrode and the pH electrode.
The preparation of the enzyme modifying solution comprises the following steps: firstly, dissolving chitosan by acetic acid to prepare a 0.1% chitosan solution, adding a multi-walled carbon nanotube and lactate dehydrogenase into the 1mL chitosan solution, and uniformly mixing to prepare the enzyme modification solution.
Said H+Preparation of selective membrane solution: mixing trilaurylamine and tetra [3, 5-bis (trifluoromethyl) phenyl]Mixing sodium borate, polyvinyl chloride and di (2-ethyl hexyl) sebacate uniformly to obtain H+Selective membrane mixture, adding cyclohexanone into the above 100mg mixture, mixing, and making into H+A selective membrane solution.
The preparation of the lactic acid sensing electrode comprises the following steps: and (3) taking a certain amount of the enzyme modified liquid to be dripped on the surface of a specified flexible gold electrode, and drying in vacuum at room temperature.
The pH electrode is prepared: taking a certain amount of the above-mentioned H+And (3) dripping the selective membrane solution on the surface of the appointed flexible gold electrode, and drying in vacuum at room temperature.
Preparing the reference electrode: placing the silver electrode on the flexible substrate in 0.1M FeCl3And (5) taking out the solution for 40s, washing the solution clean by using deionized water, and drying the solution in vacuum at room temperature.
The flexible sweat lactic acid electrochemical sensor with pH correction is used for detecting lactic acid: preparing a batch of sensors in parallel, connecting the sensors with an electrochemical workstation, respectively placing the sensors in sample solutions with different pH values, respectively detecting current signals under 0V potential by using a chronoamperometry method, then adding lactic acid with different concentrations into the sample solutions, recording the current signals, establishing standard curves (shown in figure 2) of the current signals and the lactic acid concentration under different pH values, and further establishing the standard curves of the current signals, the pH values and the lactic acid concentration, namely the lactic acid standard curve after pH correction.
The flexible sweat lactic acid electrochemical sensor with pH correction is used for detecting pH: connecting the prepared sensor with an electrochemical workstation, respectively placing the sensor in buffer solutions with different pH values, recording a voltage signal by using a timing potential method, and establishing a standard curve of the voltage signal and the pH value.
The flexible sweat lactic acid electrochemical sensor with pH correction is used for detecting lactic acid in sweat: and (3) directly covering a sweat sample on the surface of the sensor for electrochemical detection, measuring the pH value of the sweat sample by the pH electrode according to the pH standard curve, simultaneously recording a current signal of the lactic acid sensing electrode, and quantitatively analyzing the lactic acid in the sweat sample based on the measured pH value of the sweat sample and the lactic acid standard curve based on pH correction.
The apparatus embodiments described above are illustrative only, the terms and expressions employed are used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof.
Claims (6)
1. Flexible sweat lactic acid electrochemical sensor with pH correction function and method for manufacturing sameIs characterized by comprising a flexible substrate, and a lactic acid sensing electrode, a pH electrode, a reference electrode and a counter electrode which are arranged on the surface of the flexible substrate, wherein the lactic acid sensing electrode and the pH electrode are formed by respectively dripping enzyme modification liquid and H on the surfaces of two independent flexible gold electrodes+And preparing a selective membrane solution.
2. The flexible sweat lactate electrochemical sensor of claim 1, wherein the enzyme modification fluid is a chitosan solution mixed with multi-walled carbon nanotubes and lactate dehydrogenase.
3. The flexible sweat lactate electrochemical sensor of claim 1 wherein the lactate sensing electrode utilizes lactate dehydrogenase to oxidize lactate in sweat and to electronically communicate with the electrode to generate an electrical current signal.
4. The flexible sweat lactate electrochemical sensor of claim 1 wherein the pH electrode allows real-time monitoring of the pH of the sample solution.
5. The flexible sweat lactate electrochemical sensor of claim 1 wherein the reference electrode is an Ag/AgCl reference electrode shared by the lactate sensing electrode and the pH electrode.
6. The flexible sweat lactate electrochemical sensor according to claim 1, wherein lactate in sweat is quantitatively analyzed based on a pH-corrected standard curve.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210096856.8A CN114487063A (en) | 2022-01-26 | 2022-01-26 | Flexible sweat lactic acid electrochemical sensor with pH correction function |
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| CN202210096856.8A CN114487063A (en) | 2022-01-26 | 2022-01-26 | Flexible sweat lactic acid electrochemical sensor with pH correction function |
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| CN114487063A true CN114487063A (en) | 2022-05-13 |
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| CN202210096856.8A Pending CN114487063A (en) | 2022-01-26 | 2022-01-26 | Flexible sweat lactic acid electrochemical sensor with pH correction function |
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- 2022-01-26 CN CN202210096856.8A patent/CN114487063A/en active Pending
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