CN102636539A - Preparation of silk-screen printing electrode for rapidly diagnosing diabetic ketoacidosis - Google Patents
Preparation of silk-screen printing electrode for rapidly diagnosing diabetic ketoacidosis Download PDFInfo
- Publication number
- CN102636539A CN102636539A CN2012100633999A CN201210063399A CN102636539A CN 102636539 A CN102636539 A CN 102636539A CN 2012100633999 A CN2012100633999 A CN 2012100633999A CN 201210063399 A CN201210063399 A CN 201210063399A CN 102636539 A CN102636539 A CN 102636539A
- Authority
- CN
- China
- Prior art keywords
- screen printing
- test paper
- beta
- electrode
- enzyme layer
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to the field of rapidly diagnosing diabetic ketoacidosis. Electrochemical test paper is prepared by using a silk-screen printing process; and the content of beta-hydroxybutyrate in human blood is detected by current which is generated when beta-hydroxybutyrate is oxidized by an enzyme layer on the test paper, so as to judge whether the diabetic ketoacidosis exists or not. The enzyme layer is prepared from nitrocellulose, hydroxybutyrate dehydrogenase, glutaraldehyde, potassium ferricyanide, reduced coenzyme and chitosan. The preparation method of the enzyme layer comprises the following steps of: dissolving nitrocellulose, hydroxybutyrate dehydrogenase, potassium ferricyanide, reduced coenzyme and chitosan with a phosphoric buffered solution; then reacting with glutaraldehyde; dropwise adding a mixed solution on the test paper; and drying to finally obtain the enzyme layer of the electrochemical test paper for detecting the beta-hydroxybutyrate. The silk-screen printing electrode provided by the invention can be widely applied to rapid diagnosis of diabetic ketoacidosis.
Description
Technical field
The present invention is a kind of preparation of screen printing electrode test paper of quick diagnosis diabetic ketoacidosis, is specifically related to a kind of electrochemistry test paper that can detect beta-hydroxy-butanoic acid.
Background technology
Diabetic ketoacidosis (DKA): a kind of acute complications that is diabetes.It is the blood sugar acidosis that the wretched insufficiency of the insulin that causes excites that sharply raises.Ketoboidies is because diabetic's metabolic disorder when increasing the weight of, fat motion and decompose and quicken, and abundant fatty acid produces through beta-oxidation at liver.When the ketoplasia amount too much surpassed the outer autoxidation ability of liver, blood ketone body raise to increase with the eliminating of urine ketoboidies and is called ketosis.Blood ketone body further raises, and the metabolic disorder aggravation makes organic acid generate reserve alkali in the too much consumer, acidosis symptom occurs.The diabetes ketoboidies mainly is divided into 3 types of beta-hydroxy-butanoic acids, acetoacetate and acetone.Beta-hydroxy-butanoic acid accounts for 78% in ketoboidies, and takes place in early days at the diabetes ketosis, and beta-hydroxy-butanoic acid just can have obvious rising, and this moment, acetoacetate did not still have significant change.Often cause clinical underestimating so measure the nitroprusside test of acetoacetate to total ketone amount and ketosis degree; In ketosis convalescence, when beta-hydroxy-butanoic acid descended rapidly, acetoacetate still kept raising or descending slowly within a certain period of time, at this moment can cause clinical over-evaluating the state of an illness again with the nitroprusside method.Therefore, measure more sensitivity than acetoacetate in the diagnosis that is determined at the diabetes ketosis of beta-hydroxy-butanoic acid, the treatment monitoring, more reliable, also very valuable in the advance notice of diabetes control equally.
Beta-hydroxy-butanoic acid adopts the enzyme kinetics continuous monitoring method usually at present, when pH8.5, measures the speed that the reduced coenzyme absorbance rises under the 340nm wavelength, is directly proportional with the concentration of beta-hydroxy-butanoic acid.Need formulate typical curve in advance when this method is measured, so required time is long, the operation more complicated, and can not detect in real time sample.
Summary of the invention
The objective of the invention is to prepare a kind of screen printing electrode test paper of quick diagnosis diabetic ketoacidosis, have advantage quick, accurate and that can detect in real time, preparation is simple, easy realization of industrial production.
The invention discloses a kind of preparation method of screen printing electrode test paper of quick diagnosis diabetic ketoacidosis, specifically implement through following steps:
(1) printing conductive silver slurry forms electrode strip on polyester film, and 70 ℃ solidify 0.5 h;
(2) with printing ink reducer allotment carboxylated CNT and carbon slurry, filtered through gauze prevents that CNT from stopping up web plate, printing carbon paste printing ink, and 120 ℃ solidify 45 min;
(3) current-carrying part in the middle of the printing dielectric ink covers, 80 ℃ solidify 20 min (as shown in Figure 1);
(4) 1 mg cellulose nitrate, 2 mg hydroxybutyric dehydrogenases, the 2 mg potassium ferricyanides, 2 mg reduced coenzymes and 1 mg shitosan being used the pH of 10 mL is the dissolving of 7.4 phosphate buffer solution; With 5uL content is that 1% glutaraldehyde solution added in the above-mentioned mixed solution stirring reaction 0.5 hour; 10 uL drips of solution are added on the working electrode 1 45 ℃ of drying 0.5 h;
(5) 1 mg cellulose nitrate, the 2 mg potassium ferricyanides, 2 mg reduced coenzymes and 1 mg shitosan being used the pH of 10 mL is the dissolving of 7.4 phosphate buffer solution; With 5uL content is that 1% glutaraldehyde solution added in the above-mentioned mixed solution stirring reaction 0.5 hour; 10 uL drips of solution are added on the working electrode 2 45 ℃ of drying 0.5 h.
Diabetic ketoacidosis screen printing electrode of the present invention utilizes electrochemical workstation CHI660B to carry out chronoamperometry i-t test, and the response time is less than 20s.Above-mentioned test paper is 0.05 –, 5.0 mmolL in beta-hydroxy-butanoic acid concentration
-1Have good linear relationship in the scope, detection limit is 4.6 m molL
-1Because normal person's beta-hydroxy-butanoic acid concentration is lower than 0.27 mmolL
-1, therefore, this test paper can detect diabetic ketoacidosis real-time.
Description of drawings
Fig. 1. screen printing electrode test paper structure
Among the figure: 1 is base material, and 2 is silver layer, and 3 is carbon-coating, and 4 is insulation course, and W1 is first working electrode, and R is a contrast electrode, and W2 is second working electrode
Fig. 2. (beta-hydroxy-butanoic acid concentration is 0.01-10.0mmolL to the current-responsive of the concentration of beta-hydroxy-butanoic acid and screen printing electrode test paper
-1)
Fig. 3. different batches screen printing electrode test paper is to the response of beta-hydroxy-butanoic acid
Embodiment
Embodiment 1 preparation quick diagnosis diabetic ketoacidosis screen printing electrode
It is the dissolving of 7.4 phosphate buffer solution that 1 mg cellulose nitrate, 2 mg hydroxybutyric dehydrogenases, the 2 mg potassium ferricyanides, 2 mg reduced coenzymes and 1 mg shitosan are used the pH of 10 mL; With 5uL content is that 1% glutaraldehyde solution added in the above-mentioned mixed solution stirring reaction 0.5 hour; 10 uL drips of solution are added on the working electrode 1 45 ℃ of drying 0.5 h; It is the dissolving of 7.4 phosphate buffer solution that 1 mg cellulose nitrate, the 2 mg potassium ferricyanides, 2 mg reduced coenzymes and 1 mg shitosan are used the pH of 10 mL; With 5uL content is that 1% glutaraldehyde solution added in the above-mentioned mixed solution stirring reaction 0.5 hour; 10 uL drips of solution are added on the working electrode 2; 45 ℃ of drying 0.5 h obtain quick diagnosis diabetic ketoacidosis screen printing electrode.
The concentration of the beta-hydroxy-butanoic acid that embodiment 2 is different is at the current-responsive of screen printing electrode test paper
At first accurate weighing 0.198 g beta-hydroxy-butanoic acid is dissolved in 10 mL PBS solution and (contains 0.1 molL
-1KClO
4) in, concentration is 100 mmolL
-1, use the PBS solution dilution, preparation 0.01,0.03,0.05,0.06,0.08,0.1,0.3,0.5,0.8,1.0,1.5,2.0,2.5,3.0,3.5,4.0,4.5,5.0,5.5,6.0,6.5,7.0,8.0,9.0 and 10.0 mmolL
-1Beta-hydroxy-butanoic acid solution, measure the current-responsive of the beta-hydroxy-butanoic acid of variable concentrations with the CHI660B electrochemical workstation at the screen printing electrode test paper, current potential is 0.4 V, and is as shown in Figure 2.Beta-hydroxy-butanoic acid concentration is at 0.05 –, 5.0 mmolL
-1Electric current with the silk screen pole reagent paper in the scope has good linear relationship, and detection limit is 4.6 m molL
-1
At first accurate weighing 0.198 g beta-hydroxy-butanoic acid is dissolved in 10 mL PBS solution and (contains 0.1 molL
-1KClO
4) in, concentration 0.27 mmolL
-1, be that 0.4 V measures the current-responsive of beta-hydroxy-butanoic acid at the screen printing electrode test paper with the electrode of 10 different batches at current potential, as shown in Figure 3.
Claims (4)
1. the preparation of the screen printing electrode test paper of a novel detection beta-hydroxy-butanoic acid, this test paper is made up of screen printing electrode and enzyme layer two parts.
2. screen printing electrode according to claim 1 is to be formed by silver slurry, carbon slurry and dielectric ink printing, and electrode comprises two working electrodes and a contrast electrode.
3. carbon slurry according to claim 2 is obtained through printing ink reducer allotment back by CH-10 conductive carbon paste and carboxylic carbon nano-tube; The diameter of CNT is 40nm-60nm.
4. enzyme layer according to claim 1, its table preparation process comprises the following step: it is 7.4 phosphate buffer solution dissolving that the pH of 10 mL is used with 1 mg cellulose nitrate, 2 mg hydroxybutyric dehydrogenases, the 2 mg potassium ferricyanides, 2 mg reduced coenzymes and 1 mg shitosan in (1); (2) be that 1% glutaraldehyde solution added in the above-mentioned mixed solution stirring reaction 0.5 hour with 5uL content; (3) drips of solution is added on the test paper 45 ℃ of dry 0.5h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012100633999A CN102636539A (en) | 2012-03-12 | 2012-03-12 | Preparation of silk-screen printing electrode for rapidly diagnosing diabetic ketoacidosis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012100633999A CN102636539A (en) | 2012-03-12 | 2012-03-12 | Preparation of silk-screen printing electrode for rapidly diagnosing diabetic ketoacidosis |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102636539A true CN102636539A (en) | 2012-08-15 |
Family
ID=46621012
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012100633999A Pending CN102636539A (en) | 2012-03-12 | 2012-03-12 | Preparation of silk-screen printing electrode for rapidly diagnosing diabetic ketoacidosis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102636539A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103913502A (en) * | 2012-12-31 | 2014-07-09 | 北京师范大学 | Copper rapid determination method based on square-wave stripping voltammetry and three-electrode sensor |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100270175A1 (en) * | 2009-04-22 | 2010-10-28 | Nova Biomedical Corporation | Electrochemical biosensors based on NAD(P)-dependent dehydrogenase enzymes |
-
2012
- 2012-03-12 CN CN2012100633999A patent/CN102636539A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100270175A1 (en) * | 2009-04-22 | 2010-10-28 | Nova Biomedical Corporation | Electrochemical biosensors based on NAD(P)-dependent dehydrogenase enzymes |
Non-Patent Citations (4)
| Title |
|---|
| 《Sensors and Actuators B》 20070929 Lei Fang等 An electrochemical biosensor of the ketone 3-beta-hydroxybutyrate for potential diabetic patient management 第818-825页 1-4 第129卷, 第2期 * |
| G. LI等: "A new handheld biosensor for monitoring blood ketones", 《SENSORS AND ACTUATORS B》, vol. 109, no. 2, 1 February 2005 (2005-02-01), pages 285 - 290 * |
| LEI FANG等: "An electrochemical biosensor of the ketone 3-β-hydroxybutyrate for potential diabetic patient management", 《SENSORS AND ACTUATORS B》, vol. 129, no. 2, 29 September 2007 (2007-09-29), pages 818 - 825 * |
| 马念章: "酶生物传感器的研究及其在糖尿病检测中的应用", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》, no. 05, 15 September 2005 (2005-09-15), pages 1 - 65 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103913502A (en) * | 2012-12-31 | 2014-07-09 | 北京师范大学 | Copper rapid determination method based on square-wave stripping voltammetry and three-electrode sensor |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| DE102006043718B4 (en) | Determination of hydrogen peroxide concentrations | |
| Lin et al. | A facile electrochemical method for simultaneous and on-line measurements of glucose and lactate in brain microdialysate with prussian blue as the electrocatalyst for reduction of hydrogen peroxide | |
| TWI450967B (en) | Homogeneously-structured nano-catalyst/enzyme composite electrode, fabricating method and application of the same | |
| Moreno et al. | Selective detection of dopamine in the presence of ascorbic acid using carbon nanotube modified screen-printed electrodes | |
| CN107037102B (en) | Nano composite material and preparation method and application thereof | |
| CA2776332C (en) | Underfill recognition system for a biosensor | |
| Wang et al. | Sensitive electrochemical detection of cholesterol using a portable paper sensor based on the synergistic effect of cholesterol oxidase and nanoporous gold | |
| CN111366625B (en) | Preparation method of ratio electrochemical uric acid sensor based on zinc coordination black phosphorus nanosheet compound and biological enzyme catalysis | |
| US20190046092A1 (en) | Three-dimensionally printed blood glucose sensing device and fabrication method | |
| CN103235015A (en) | Dry chemical test strip for quantitatively testing uric acid | |
| CN1142867A (en) | Methods and apparatus for electrochemical measurements | |
| CN111351828B (en) | ZnFe 2 O 4 Preparation method and application of modified electrode material | |
| CN108776163B (en) | Enzyme biosensor for detecting cholesterol and preparation method and application thereof | |
| US20240252072A1 (en) | Treatment device using reaction system containing oxidoreductase and co-enzyme, and method for controlling same | |
| Bahcegul et al. | Selecting the right blood glucose monitor for the determination of glucose during the enzymatic hydrolysis of corncob pretreated with different methods | |
| Kumar et al. | Electrochemical cholesterol sensors based on nanostructured metal oxides: Current progress and future perspectives | |
| CN115097149A (en) | Method for simultaneously detecting glucose and cholesterol, electrode and electrode manufacturing method | |
| CN103267783B (en) | A kind of method being detected hydrogen oxide, uric acid or uriKoxidase | |
| CN102636539A (en) | Preparation of silk-screen printing electrode for rapidly diagnosing diabetic ketoacidosis | |
| Lin et al. | Nickel hexacyanoferrate modified screen-printed carbon electrode for sensitive detection of ascorbic acid and hydrogen peroxide | |
| Malik et al. | Construction of an amperometric pyruvate biosensor based on enzyme bound to a nanocomposite and its comparison with enzyme nanoparticles bound to electrode | |
| Zhang et al. | Online electrochemical systems for continuous neurochemical measurements with low-potential mediator-based electrochemical biosensors as selective detectors | |
| CN111796016A (en) | Method for detecting glucose by electrochemical/colorimetric dual-mode sensor | |
| JPWO2005056824A1 (en) | Method for detecting sulfate-conjugated bile acid, test paper and biosensor used therefor | |
| CN116789106B (en) | Terpyridine ruthenium doped mesoporous carbon nanosphere and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20120815 |