TW200306416A - Electrical potential type urea sensing device and the manufacturing method thereof - Google Patents

Abstract

The present invention discloses an electrical potential type urea sensing device and the manufacturing method thereof, wherein a conductive layer is first formed on the substrate, then deposit a sensing film as the ion sensing electrode, the sensing film has a sensing region and a non-sensing region; use a conductive wire to extend from the conductive layer as the electrical contact with the exterior; encapsulate the non-sensing region of the sensing film by an encapsulant, define a sensing window in the sensing region; finally, use enzyme immobilization technology to immobilize the urea enzyme on the sensing window in the sensing film, so as to complete the fabrication of an electrical potential type urea sensor device. The present invention is a disposable urea sensing device, and has the functions of mass duplication, low cost, and ease of packaging.

Description

200306416 V. Description of the invention (l) 1. [Technical field to which the invention belongs] The present invention relates to a potential-type urea sensing element in an acid-base sensing film using tin dioxide as an extended off-field effect (EGFET) A technician who works with urea enzymes to make urea sensing elements. / ~ Press, and the correlation is related to the index points, such as continuous flow, roads, and materials. The biological release sensing has field elements and points. Sensor-fixed sensors Prior technology] ‘Simultaneously in the blood and kidney’ So blood is used in clinical diagnosis. Traditionally, there is a kind of biological material science and light sensor, which is complicated in operation and tested by Cheng Zhizheng. Sub-sensing field effect element, this element effect transistor has been miniaturized. It is used in quantification of urea concentration, liver function, or urine interruption, which is called 'in 1980 in ion sensing'; therefore, The principle of the sensor, science and other systems is the response energy and renal urea nitrogen. The kidney analysis method is too long. In order to make up for its binding, the concentration function in the protein adrenal gland is set to the actual equipment. Ingestion and alienation for secretory function have important parameters closely related to human health. There are many shortcomings in the application, which are not suitable for quantitative analysis. The electronic crystal system required in various fields of chemistry and electronics, which appeared in 1970 and developed rapidly, has the function of an ion-selective electrode and also has characteristics. It is completely different from traditional electrodes; this type of sensing is easy to be instrumented and suitable for the design of automated systems. Car as and Janata first used the gate of penicillin as a field-effect transistor for enzyme sensing. .

200306416 V. Description of invention (2) Since its development, many related patents have been proposed, such as US Patent No. 5,992, 183, 'Metal oxide matrix biosensors', which discloses a metal oxide film as a biomolecule The substrate is provided, and at the same time, a current type or a potential type biosensor is provided, and the enzyme, antibody, antigen, and nucleic acid sequence are used for sensing. U.S.A., No. 5,858,186, Urea biosensor for hemodialysis

monitoring " is an electrochemical sensor that can quantitatively detect the concentration of urea by using the dialysis waste liquid during hemodialysis. This sensor uses enzymes to hydrolyze urea and detect the pH value of the urea produced. The change. US Patent No. 5, 833, 824, Dorsal substrate guarded ISFET sensor, which uses an ion-sensing field effect transistor to sense the ionic activity of a solution. U.S. Patent No. 4,877,582, Chemicai deViCe with effect transistor ", which is a field-effect transistor as an enabler, and uses a suitable biometric identifier i for a substance to be detected, and then uses the field-effect transistor. The crystal converts the chemical signal Γ into an electrical signal so that the specific substance in the solution can be quantitatively detected.

Urea in urine is widely used, and the present invention is to produce ten pieces in combination with semiconductor process technology for the blood acupoint sensing element chen t 'Ik to make it-to detect the pH value as Basic biosensing 5 'Structures that can be adapted to be developed as disposable sensing elements ^ 2. [Summary of the invention] The main purpose of the present invention is to provide a potential-type urea sensing,

200306416 V. Description of Invention (3)

And its manufacturing method ', which uses a non-insulating solid-state ion-sensing film as the sensing electrode of an ion-sensing field-effect transistor, and combines semi-process technology to produce a disposable urea sensing element. Γ μi invention: In addition, the purpose is to provide a potential-type urea sensing element and a method for making clothes. The eight series can be reproduced in large quantities and has the characteristics of low cost and closeness, so it can reduce the conventional ion sensing field. Packaging issues. The cost and interstitialization of Wendong

Still another object of the present invention is a method and a method for manufacturing the same. The method has the advantages of easy storage, packaging, and adjustment points, and provides a potential-type urea sensing element that is simple, low-cost, and capable of drying the sensing area and transportation. Convenience, etc. One embodiment of the present invention is a structure of a potential-type urea sensing element, which is provided with a sensing film on a substrate having a conductive layer on the surface? It is used as an ion sensing electrode, and the sensing film has a sensing area and a non-sensing area; and a conductive line is extended from the conductive layer. The encapsulating gel covers the non-test area of the sensing film, and defines a sensing window of the sensing area; and a urease, which is fixed in the sensing window on the sensing film to make use of the urea sensor. The measuring element detects the urea concentration in blood or urine, and serves as a judgment parameter for clinical interpretation. Another embodiment of the present invention is a method for manufacturing a potential-type urea sensing element. The steps include: first providing a substrate, and then forming a conductive layer on the surface; and forming a sensing film on the surface of the conductive layer of the substrate. So that ^ is an ion sensing electrode, and the sensing film has a sensing area and a non-wei

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Then, a non-sensing area on the sensing film is formed and a sensing window is formed; and finally, the sensing window on the sensing film is used to solidify the sensing window as described above. It can also be used to sense the known urea concentration. The conductive line extending from the layer is used as a packaging step. A packaging colloid encapsulation field is used to define the sensing area. A urea enzyme is fixed there, and a urea and a sensing element can be completed. The detection of the base number, and then the following measures are given by the specific examples in conjunction with the description of the diagram attached to ία 4 σ, and it will be easier to understand the purpose of the present invention when it is more useful. + Zhiyue's technical content, characteristics and achievements. [Implementation] This hybrid system uses tin dioxide as ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ I ^ rr; ^ ^ ^ θ 髀 .〇 · 加冰 A «It is said that the self-extending gate ion sensing field is applicable to various types of sensing wood structures for detection. H-based life ::: is a potential-type urea sensing element 10, such as made of-indium tin oxide (IT0) conductive two glass two substrates two 'surface system ion sensing old amine 1 g 丄-sheet Injustice 14, there is another non-insulating solid tin conductive reed 1'4 :: 5 solid materials such as tin, which are located in indium oxide acid as a solid ion sensing electrode ... Has a sensing area and a non-sensing =

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Domain · 'Reuse a conductive wire 18 extending from the indium tin oxide conductive layer 丨 4 to serve as a signal transmission line and external electrical contacts; and a packaging gel 20 covering the non-sense of the sensing film 16 A sensing area, and a sensing window 22 is defined to define the sensing area 10 of the sensing element 10 using the encapsulant 20 to be about 2 * 2 mm2; and a urease 24 is fixed to the sensing area Inside the edge window 22 on the film 16, and the urease 24 is urea embedded in pVA_SbQ colloid? Composed of. This urea sensing element is 10 times better than the conventional ion-sensing field-effect transistor. It is easier to make a seal ^ ~ Θ ^ ^ " Your clothes F, easier on the table, and can reduce the cost to fit and discard Condition of the sensing element. Among them, the above-mentioned potential-type urea substrate is used as an element substrate, and the other substrates are non-insulating substrates, and are insulating or polymer substrates. Due to the variability of the plates, different materials can be used for practical applications. The embodiment of the sensing element uses a glass substrate. The substrate can also be selected from other insulating base substrates. The substrate is selected from silicon substrates and ceramic substrates. This sensing element has better basic conditions and process conditions. The method of broadcasting in the first picture shown in the first figure above will explain the manufacturer of the present invention with M, gate < ,,, and mouth structure. Δ Yuemai reads the second (a) figure to the second example The manufacturing method of the potential-type urea sensing element j is shown in the preferred embodiment of the present invention, and the manufacturing method of the present invention includes a cross-sectional view of the structure of the structure; as shown in the first step, please refer to the second step u). The following steps: and A glass substrate 12 is provided at JL it .. A glass substrate 12 is provided on the upper surface. An emulsion indium tin guide 14 is formed on the upper surface. The argon layer 9 is a β layer 14. The thickness of the indium tin oxide conductive layer 4 is 230 angstroms. And the resistance is 50˜100. Then, the fatigue a on the glass substrate 12 is exhausted. Electroplated growth-the surface of the indium tin conductive layer 14 of the layer of tin dioxide is used to bind the film 16 as shown in the second (b)

200306416 V. Description of the invention (6) Do not 'this step is a mixed gas of 2) and-oxidation "'," pass in lice and oxygen (4 · · beautiful 柘 1? Less, w ώ and an emulsion The sensing film 16 of tin grows, and the temperature of the glass substrate 12 is maintained at j:% glass as solid ions 4: Π; the thin group 6 with a thickness of 2000 angstroms, with the area and non-formation S t, This sensing film 16 series can be provided with a conductive wire 181 for sensing 锢 tin conductive 系. Oxidized mn is used as an external electrical contact. The encapsulation gel such as grease, etc. is sealed with epoxy resin. In order to define the sensing area—the corpse on the sensing window #: As shown in the first figure, using the enzyme fixing technology, the sensor window is fixed on the sensing film 16. Within 22; In this, ΐ: Γ series! Utilizing the characteristics of photopolymerization and photopolymerization, the urea enzyme 24 is solidified 2; the second test ® 2 2 functions to complete the urea sensor element i can be used. Other immobilized systems that can form potential-type electric / dagger students: sense: device 'in order to reduce the cost of large-scale instruments such as optical biological analysis It is suitable for real-time detection and disposal of discarded components.. Sheep's enzyme fixation technology. Read the following instructions: First, prepare urea f (urease, EC 3.5.1.5, 50000 ~ 1 0 0 0 0 0 units / g); PVA-SbQ colloid, where pvA (Dp II Π00, DS · II 88), SbQ (1 · 25 mol%), N v = 12 ⑼ wt%, PH is 5.6, in The viscosity at 25 ° C is about 5750 cp; urea (NH2CONH2 = 60.06), 9%; and phosphate (KH2po4z; 200306416) 5. Description of the invention (7) 1 3 6 · 09), As a general reagent grade, it is used for the configuration of a buffer solution. Next, PVA-SbQ is diluted (100 mg / 100 microliters of 55 millimolar pH value 7.0 phosphate solution) and an enzyme solution (7 mg / 100 microliters of mololeic acid with a salt filling solution of 7.0 · 0) After mixing at a ratio of 1: 1, take 1 microliter of drops onto the sensing window 22, and then 'send the sensing element factory 〇 Place under 4W, 365nm UV light, fill in the photopolymerization reaction for about 20 minutes. Finally, after the photopolymerization reaction is over, place the sensing element in a dark box at 4 ° C. It takes about 12 hours The process of enzyme immobilization is completed. As shown in the third figure, it is a potential-type urea sensing element 10 with a separated tin dioxide / indium oxide / glass substrate structure as a transducer, and it uses conductivity Line 1 8 is electrically connected to a potentiometer with high input impedance, such as a metal half field effect transistor and an operational amplifier. The readout circuit in this figure is Lt 11 6 7 instrumentation amplifier 2 6 with silver / gasified silver electrode 2 8 A reference stable potential is provided to measure the reaction voltage of the urea sensing element in the solution, and then calculate the solution concentration. The fourth figure is the pH correction curve of the urea sensing element of the present invention. As shown in the figure, it is the acid-base sensing characteristic of the tin dioxide sensing film. Within a stable range; different voltage responses caused by the above-mentioned instrumentation amplifier readout element in different pH environments. Among them, between pH 2 · 2 ~ 10 · 2, the sensitivity of the tin dioxide sensing film is about 58 · 85 ± 0 · 41 mV / pH. The fifth figure is a state diagram of the response time and the recovery time of the urea sensing element of the present invention. As shown in the figure, it shows the time required for the urea sensing element to respond and the time required for recovery. It can be known from the experimental results that

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V. Description of the invention (8) After 60 seconds, the element's response has reached g 0% of the maximum reaction voltage, and then the element is returned to the 5 millimolar buffer solution, and after about} 0 minutes or so, its The response voltage will slowly return to the voltage before the reaction. Because the purpose of this extended sensing element architecture is to develop a disposable biosensor, the length of the recovery time is It is not necessary to consider parameters. The sixth figure is the urea sensing element = calibration curve diagram of the present invention in the measurement environment of different acid-base values, which is the effect of the reaction of the test solution and the calibration curve when the solution to be measured changes. . ： = The more alkalinity value f3 temples, the range of its reaction can be significantly reduced; the hydrolysis reaction: the maximum changeable limit is acid test value 9 ·:;, if the initial pH value of urine production fluid is too high, The reaction can be changed. The test bath is small; on the contrary, if the relative activity of the test solution is affected, the same result will be J: 夂 will find an equilibrium point between the two enzymes. —I 殳 h, so how can 6.0 be the best reaction environment? From this test result, you can know that the pH value is shown in the seventh figure, which is the calibration of the urea sensing element of the invention. Qu§ _ _, 'The influence of the bath concentration on the reaction, because of the influence of two :::: different buffer solution concentrations to the buffering capacity of the buffer solution Π: species: j :, the detector is susceptible to this The strength of the buffer solution has a strong relationship with the concentration of Γ ς j. The concentration of the buffer solution can be changed === force = can change, so its calibration curve will be the voltage 220064016 5. Explanation of the invention (9) The eighth figure is The increase of urea sensing element of the present invention over time = a schematic diagram of the change of the maximum reactable voltage. 2: The stability of the existence of the sadness is to prepare 100 in advance to extend 4 indium tin oxide / glass ion sensing field effect transistor to make tin / and save the completed sensor in 4. 仏 dark box Perform a measurement once, observe how long the sensor can be stored after the production is completed. From the experimental results, it can be seen that the sensor can work properly when it is over 0, feel old, and have a hole. The device still has no obvious attenuation phenomenon. Therefore, the present invention uses a hybrid structure of indium tin / glass substrate: π, βί π type of emulsified tin / oxygen salt solution The device has a acid value of 5 millimolar at 6 and a concentration of 0.31 mg / 100 milliliter. The soil response curve 'can be used to detect the sensitivity of urea. The linear part of the range of "millivolt / ^ 100 liters'," Tex / ρ [urea concentration]. Sensing field effect electricity: 2: Ion sensing film is used as ions to make disposable urinals and = right conductor process technology to produce low-mold and easy-to-package features. It has the cost and simplification of a large number of replicable and electric crystals. The low field ionization field effect of 17 cows, such as packaging, has the advantages of simple production, low cost, and x1. In addition, the present invention also has an integrated sensing area, and is easy to be packaged for transportation and storage. The above-mentioned embodiment is only an advantage of 2 γ. The purpose is to familiarize yourself with this technology and to implement the technical ideas and features of the present invention. If you ca n’t understand the scope of the patent that defines the invention within the scope of the invention, that is, Page 200306416 V. Description of the Invention (ίο) Any equivalent change or modification made according to the spirit disclosed by the present invention shall still be covered by the patent scope of the present invention. Description of drawing number: 10-position type urea sensing element / 12 glass substrate 14 Λ indium tin conductive layer 16 sensing film 18 conductive wire 20 packaged gel 22 sensing window 24 urea enzyme 26 instrument meter amplifier 28 silver / gas silver electrode

Page 15 200306416 Brief description of the drawings The first figure is a cross-sectional view of the structure of a potential-type urea sensing element of the present invention. Figures 2 (a) to 2 (d) are cross-sectional views of the steps of the present invention in the steps of manufacturing a potential-type urea sensing element. η ~ The third figure is a measurement architecture diagram of the urea sensing element of the present invention. The fourth figure is a curve diagram of the calibration curve of the urea sensing element of the present invention. The fifth graph is a response time and recovery time state diagram of the urea sensing element of the present invention. The sixth figure is a graph of the calibration curve of the urea sensing element according to the present invention when the environment is measured at different I base values. The seventh figure is a graph of the calibration curve of urea sensing elements with different buffer solutions. The eighth figure is a graph showing the change of the largest reactable electrical waste that urea f = element of the present invention shows over time.

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Claims (1)

200306416 6. Scope of patent application 1. A potential-type urea sensing element, including a substrate having a conductive layer formed on its surface; a sensing film located on the surface of the conductive layer of the substrate and serving as an ion sensing electrode, the sensor The test film has a sensing area and a non-sensing area \ 々a conductive line, which is extended from the conductive layer as an external contact; ^ a packaging gel, which covers the sensing A non-sensing area of the film, and defining the sensing area as a sensing window; and A urea enzyme is fixed in the sensing window on the sensing film. (1) The potential-type urea sensing element according to item 1 of the patent application, wherein the substrate is selected from an insulating substrate and a non-insulating substrate. -3. The potential-type urea sensing element according to item 2 of the scope of the patent application, wherein the insulating substrate is selected from the group consisting of a silicon substrate, a glass substrate, a ceramic substrate, and a high-knife polymer substrate. The conductive layer is indium tin oxide as described in item 1 of the scope of patent application. 5 > In the potential-type urea sensing element described in item 1 of the% patent scope, the sensing film is a non-insulating solid-state ion sensing film. 1 \ The material of the sensing film in the potential-type urea sensing element described in item 1 of the scope of patent application is tin dioxide. V. Please refer to the potential-type urea sensing element described in item 1 of the above range. ^ The sensing film is formed by a sputtering method. 8. Potential as described in item 丨 of the scope of patent application "Wires provide potentiometers connected to -high wheel resistance. Page 17 200306416 6. Scope of patent application 9. The potential-type urea sensing element as described in item 1 of the scope of patent application, wherein the urease is fixed in the sensing window using enzyme fixation technology. 10. The potential-type urea sensor according to item 1 of the scope of the patent application, wherein the urease is urea embedded in a PVA-SbQ colloid? Composed of. 11. A method for manufacturing a potential-type urea sensing element, which includes the following steps: providing a substrate on which a conductive layer is formed; forming a sensing film on the surface of the conductive layer of the substrate as an ion A sensing electrode, the sensing film has a sensing area and a non-sensing area, a field; forming a conductive line extending from the conductive layer as an external electrical contact; performing a packaging step, the system Encapsulating the non-sensing area of the sensing film with an encapsulating gel, and defining a sensing window for the sensing area; and fixing the urea enzyme to the sensing film using the enzyme fixing technology Inside the window. 12. The manufacturing method as described in item 11 of the patent application scope, wherein the substrate is selected from an insulating substrate and a non-insulating substrate. 13. The manufacturing method according to item 12 of the scope of the patent application, wherein the insulating substrate is selected from the group consisting of a silicon substrate, a glass substrate, a ceramic substrate, and a polymer substrate. 14 layers: The manufacturing method as described in item Π of the patent application scope, wherein the conductive system is indium tin oxide. The manufacturing method described in item n of the scope of patent application, wherein the sensing Page 18 200306416 6. Scope of patent application The film is a non-insulating solid-state ion sensing film. 16. The manufacturing method as described in item 11 of the scope of patent application, wherein the material of the sensing film is tin dioxide. 17. The manufacturing method as described in item 11 of the scope of patent application, wherein the sensing film is formed by a sputtering method. / '1 8. The manufacturing method as described in item 11 of the scope of patent application, wherein the urea enzyme is fixed in the sensing window by means of physical embedding. 19. The manufacturing method as described in item 11 of the scope of the patent application, wherein the urea enzyme is composed of urea f embedded in the PVA-SbQ colloid. - Page 19