TW201409026A - Potentiometric lactate senor, method for forming thereof and measuring system including thereof - Google Patents

Abstract

A potentiometric lactate senor is provided, in which includes a substrate, a lactate sensor array and a conductive layer. The lactate sensor array includes a plurality of lactate sensor arranged on the substrate and isolated to each other by an isolation layer. Each the lactate sensor includes a semiconductor sensing layer on the substrate and an enzyme layer on the semiconductor sensing layer. The enzyme layer includes a catalyzingenzyme and electron transporting substance. The conductive layer includes an electrode array disposed in the lactate sensor array. In addition, a method for forming the potentiomeric lactate senor and a measuring system containing the potentiomeric lactate senor are also provided.

Description

Voltage type lactic acid sensor, manufacturing method thereof and measuring system therewith

The present invention relates to biomedical sensors, and more particularly to a voltage type lactic acid sensor and a method of manufacturing the same.

Lactic acid is a product of glycolytic hydrolysis of glucose, which is widely present in living organisms and can be applied as a detection target for biosensors. For example, in the clinic, blood and lactic acid and pyruvate content are often used to indicate that glucose in tissues is the standard for aerobic or anaerobic glycolysis, and is used as a basis for judging the oxygen supply of tissues. In general, the concentration of lactic acid in the serum of normal people and patients is different, and the concentration of lactic acid after athletes undergo anaerobic exercise will also increase significantly. In the food industry, the lactic acid concentration can also be used as a basis for the interpretation of the quality of dairy products, wine, and fruits and vegetables.

The concentration of lactic acid can be detected by various sensing elements. Piet Bergveld first developed the technology of ion-sensing field-effect transistor (ISFET) detection as a lactic acid concentration sensing element. The ion-sensing field-effect transistor is similar to the usual gold-oxygen half-field effect transistor (MOSFET). The main difference is that the ion-sensing field-effect transistor is replaced by a cerium oxide insulating layer and an electrolyte to replace the gold-oxygen half-field effect transistor. Metal gate. By sensing the characteristics of the adsorption bond formed by the membrane and the ion to be tested in the buffer solution, the potential change on the surface of the sensing membrane is measured, and then the concentration of lactic acid in the solution is detected.

Next, there are many techniques for improving ion sensing field effect transistors. For example, using yttrium oxide as a sensing film to self-test the ion sensing film The field effect transistor in the solution is separated to form an extended gate field effect transistor to improve the ion sensing field effect transistor is susceptible to light, and to solve the ion sensing field effect transistor It is not easy to package under the standard process of gold oxide half field effect transistor. Furthermore, the extended ion sensing field effect transistor has also been modified to be suitable for use in disposable sensing elements, which not only avoids direct contact between the electrical regions of the device and the aqueous solution, but also can be used in standard processes with gold oxide half field effect transistors. The technology is fully compatible.

At present, the instrument for detecting the concentration of lactic acid can be generally a current sensor, which detects the current subjected to the redox reaction on the surface of the electrode, but has the disadvantage of being susceptible to interference from other ions. The voltage sensor is a sensor using another mechanism, which is to detect the change of the pH value caused by the chemical reaction of the buffer solution, and can modify the electrode by using the enzyme and the polymer film to generate the pair. The characteristics of lactic acid selection. However, even with voltage sensors, there are still phenomena of interference by other ions, and regardless of the mechanism, the current price of instruments for detecting lactic acid concentration is still very high, although it is widely used.

Based on the current development trend of the biomedical industry, what is needed is a detector that enables real-time monitoring of patients in home testing. Therefore, sensing elements that are miniaturized, low-cost, and highly selective are development goals of the biomedical industry.

Based on the above, the present invention provides a voltage type lactic acid sensing element prepared in a separate extended ion sensing field effect transistor structure.

In an embodiment, the present invention provides a voltage lactic acid sensing The device comprises: a substrate; an array of lactic acid sensing elements, comprising a plurality of lactic acid sensing elements arranged on the substrate and isolated from each other by an isolation layer, wherein each lactic acid sensing element comprises: a semiconductor sensing layer On the substrate; and an enzyme layer on the semiconductor sensing layer, having an exposed upper surface, wherein the enzyme layer comprises a catalytic enzyme and an electron transporter; and a wire layer comprising an electrode array at the lactic acid sensation Measure the array of components. Furthermore, the present invention also provides a method of manufacturing the above-described voltage type lactic acid sensor and a measuring system including the above-described voltage type lactic acid sensor.

In another embodiment, the present invention also provides a method for manufacturing a voltage type lactic acid sensor, comprising: providing a substrate; forming a semiconductor sensing layer on the substrate, wherein the semiconductor sensing layer comprises a plurality of sensing layers Forming a wire layer on the substrate, the wire layer comprising an electrode array at least partially covering the sensing patterns; forming an isolation layer covering the substrate and exposing the sensing patterns; and forming an enzyme layer on the substrate These exposed sensing patterns form an array of lactic acid sensing elements, wherein the enzyme layer comprises a catalytic enzyme and an electron transporter.

In still another embodiment, the present invention further provides a measurement system including a lactic acid sensor, comprising: the foregoing voltage type lactic acid sensor; a reference electrode; a measuring tank, providing a solution to be tested, A voltage type lactic acid sensor and the reference electrode are placed therein; and a computer device is electrically connected to the voltage type lactic acid sensor.

The above and other objects, features and advantages of the present invention will become more <RTIgt;

The embodiments of the present invention are described in detail below with reference to the illustrated drawings. Wherever possible, the same reference numerals are in the In the drawings, the shapes and thicknesses of the embodiments may be exaggerated to make the illustrations clear and understandable. This description will describe the formation of particular components in accordance with the present invention, which may be part of the device or used directly in conjunction with the device. In the present specification, the phrase "an embodiment" means a specific element, structure or feature relating to the embodiment, and may be included in at least one embodiment. Therefore, when the phrase "in an embodiment" is used throughout the specification, it is not necessary to refer to the same embodiment. Furthermore, the particular elements, structures, or characteristics in one or more embodiments can be combined in any form. It should be noted that the illustrations shown below are for illustrative purposes only and are not shown in the original scale.

Referring to Figures 1 and 2, each of which shows a cross-sectional view and an exploded view of a voltage type lactic acid sensor in accordance with an embodiment of the present invention. In one embodiment, the voltage type lactic acid sensor provided by the embodiment of the present invention is a voltage array type lactic acid sensor.

The voltage type lactic acid sensor 100 includes a substrate 102 on which an array of lactic acid sensing elements can be disposed. The substrate 102 can be, for example, a plastic or glass substrate such as polybutylene terephthalate (PET), insulating glass, conductive glass, or a combination of the foregoing. The lactic acid sensing element array is formed by a combination of a plurality of lactic acid sensing elements 104, and these lactic acid sensing elements 104 are separated from one another by an isolation layer 106. That is to say, each of the lactic acid sensing elements 104 is surrounded by the isolation layer 106 when viewed from above. The isolation layer 106 can be, for example, a ring Oxygen resin, UV glue or waterproof insulating colloid.

In an embodiment, each lactic acid sensing element 104 can include a semiconductor sensing layer 108 and an enzyme layer 110. The semiconductor sensing layer 108 can be disposed on the substrate 102. The semiconductor sensing layer 108 can comprise germanium dioxide, tin oxide, aluminum oxide, aluminum nitride, zinc oxide, titanium oxide, graphite, combinations of the foregoing, or other semiconductor components suitable for sensing potential changes. The enzyme layer 110 can be disposed on the semiconductor sensing layer 108 and has an exposed surface to be in contact with the solution to be tested. The enzyme layer 110 can comprise a catalytic enzyme and an electron transporter. The catalytic enzyme may be, for example, lactate dehydrogenase, lactate oxidase, or other enzyme that catalyzes the redox reaction of lactic acid. The electron transporter can be, for example, potassium ferricyanide, potassium hexacyanoferrate, tetrathiafulvalene, tetracyanoquinodimethane, combinations of the foregoing. Or other materials that increase electron transport. In one embodiment, the catalytic enzyme is from about 0 mg to 5 mg, the electron transfer material is from about 0 mg to 3 mg, and the optimal weight ratio is 5:2. In addition, the enzyme layer 110 may further include a coenzyme such as nicotinamide adeine dinuleolite (LAD ⁺ ), flavin-adenine+dinuoleotide (FAD), yellow. Flavin mononucleotide (FMN), combinations of the foregoing, or other materials that aid in the catalytic reaction of an enzyme. In one embodiment, the catalytic enzyme is from about 0 mg to 5 mg, the coenzyme is from about 0 mg to 5 mg, and the optimal weight ratio is 1:1.

In one embodiment, a fixed layer 112 may be included between the enzyme layer 110 and the semiconductor sensing layer 108. The pinned layer can be, for example, a carrier 112 that simultaneously forms a covalent bond with the enzyme layer 110 and the semiconductor sensing layer 108 to The enzyme layer 110 is fixed to the semiconductor sensing layer 108. In one embodiment, the support 112 can be, for example, 3-glycidoxypropyltrimmethoxysilane (GPTS), glutaraldehyde, perfluorosulfonic acid (nafion), combinations thereof. Or other materials that can effectively immobilize enzymes.

The voltage lactic acid sensor 100 can also include a patterned conductive layer 120 disposed on the substrate 102 to transmit signals detected by the lactic acid sensing elements 102 to an external circuit. The patterned wire layer 120 can include an electrode array 122, a wire region 124, and a contact region 126. The electrode array 122 can be positioned between the semiconductor sensing layer 108 of the lactic acid sensing element and the enzyme layer 110 in contact with the enzyme layer 110. In an embodiment, the electrode array 122 can include a plurality of electrode units, each of which is correspondingly disposed in a lactic acid sensing element 104 and can serve as a working electrode of the lactic acid sensing element 102. The contact region 124 can be located outside the region of the array of lactic acid sensing elements and includes a plurality of exposed contact points for output as a voltage lactic acid sensor, providing other circuit processing devices to process the array of lactic acid sensing elements. Signal. In one embodiment, each contact point in the contact region 126 can correspond to an electrode unit located in a lactic acid sensing element. The wire region 124 of the patterned wire layer 120 is buried under the isolation layer 104 and extends between the electrode array region 122 and the contact region 126 to electrically connect the wires. In an embodiment, the patterned wire layer 120 may comprise silver paste, carbon glue, copper glue, aluminum glue, silver ink, or other suitable conductive elements.

3A to 3C are cross-sectional views showing various methods of manufacturing a voltage type lactic acid sensor according to an embodiment of the present invention. In the present embodiment, the same or similar elements as the foregoing embodiments are given the same or similar reference numerals. No. Referring to Figure 3A, a substrate 102 is first provided. The substrate 102 can be, for example, a plastic or glass substrate such as polybutylene terephthalate (PET), insulating glass, conductive glass, or a combination of the foregoing. A semiconductor sensing layer 108 may be formed on the substrate 102. The semiconductor sensing layer 108 can be formed by a radio frequency sputtering method, an evaporation method, a chemical water bath deposition method, or a combination thereof. For example, RF sputtering can be performed for about 3 to 10 minutes at a power of 100 to 120 W and 10 to 15 mTtorr. The semiconductor sensing layer 108 may form a complex sensing pattern 108 as shown in FIG. 3A via a patterning process.

Next, referring to FIG. 3B, a patterned wiring layer 108 can be formed on the substrate. The patterned conductive layer 120 can include an electrode array, a lead region, and a contact region (see FIG. 2). The electrode array can include a plurality of sensing electrodes correspondingly formed on the sensing pattern 108, and the contact region 126 A plurality of contact points may be formed outside the area where the sensing pattern 108 is located, and the wire area includes a plurality of wire connection electrode arrays and a contact area. The patterned wire layer 120 can be formed by screen printing or other suitable method.

Next, referring to FIG. 3C, an isolation layer 106 is formed on the substrate 102 to isolate the sensing patterns 108 from each other. In addition, the isolation layer 106 can substantially cover the wire regions of the patterned wire layer 120 and expose the contact regions of the sensing patterns 108 and the patterned wire layers 120. The isolation layer 106 can comprise, for example, an epoxy, UV glue, a waterproof insulating glue, an insulating protective glue, or the like, and can be formed by screen printing or other suitable method. In one embodiment, the isolation layer 106 may have a higher thickness than the semiconductor sensing layer 108 to form an opening 130 over the sensing patterns 108 for subsequent immobilization of the enzyme layer thereon.

Finally, the formation of the enzyme layer 110 forms an opening 130 above the sensing patterns 108 to form a voltage type lactic acid sensor 100 as shown in FIGS. 1 and 2. The enzyme layer 110 can comprise a catalytic enzyme and an electron transporter. The catalytic enzyme may be, for example, lactate dehydrogenase, lactate oxidase, or other enzyme that catalyzes the redox reaction of lactic acid. The electron transporter can be, for example, potassium ferricyanide, potassium hexacyanoferrate, tetrathiafulvalene, tetracyanoquinodimethane, combinations of the foregoing. Or other materials that increase electron transport. In one embodiment, the lactate is from about 0 mg to 5 mg, the electron transfer material is from about 0 mg to 3 mg, and the optimal weight ratio is 5:2. In addition, the enzyme layer 110 may further comprise a coenzyme, such as nicotinamide adeine dinuleolite (LAD ⁺ ), flavin-adenine dinucleotide (FAD), flavin. A flavin mononucleotide (FMN), a combination of the foregoing, or other material that aids in the catalytic reaction of an enzyme. In one embodiment, the catalytic enzyme is from about 0 mg to 5 mg, the coenzyme is from about 0 mg to 5 mg, and the optimal weight ratio is 1:1.

In one embodiment, the enzyme layer 110 can be attached to the sensing patterns 108 by covalent bonding. For example, a covalent bond can be generated simultaneously with the enzyme layer 110 and the sensing pattern 108 by a carrier 112. The support may, for example, be 3-glycidylpropyltrimethylnonane, glutaraldehyde, perfluorosulfonic acid (Nafion), or other material effective for immobilizing the enzyme. In one embodiment, the volume ratio of the support to the catalytic enzyme may range from 1:2 to 4:2 with an optimum volume ratio of 1:1. Alternatively, the enzyme layer can be immobilized on the sensing layer by other methods, such as cross-linking or embedding. It should be noted that although the electrode array of the patterned wire layer 120 has a portion located on the sensing pattern, the surface occupied by the electrode layer 120 The product is not large enough to affect the enzyme layer 110 to be attached to the sensing pattern 108.

In addition, in an alternative embodiment, after the enzyme layer 110 is fixed on the sensing patterns 108, the lactic acid sensor 100 can be placed in a refrigerator at 0 ° C ~ 6 ° C (preferably 4 ° C). It is stored for more than 24 hours to completely react the enzyme with the support, and is effectively covalently bonded to the sensing membrane. Thus, the enzyme layer 110 is covalently bonded to the sensing pattern 108 and maintains the activity of the enzyme.

Referring to Figure 4, there is shown a measurement system 400 including a voltage lactic acid sensor in accordance with an embodiment of the present invention. The measurement system 400 includes a lactic acid sensor 100, a measuring tank 410, a reference electrode 420, a readout circuit 430, and a computer device 440. The lactic acid sensor 100 may be a voltage type lactic acid sensor 100 as described in FIGS. 1 and 2.

When the measurement system 400 is operated, the solution to be tested 405, the lactic acid sensor 100, and the reference electrode 420 may be placed in the measurement tank 410. The solution to be tested may comprise dissolving the lactic acid analyte in a buffer solution, which may be phosphate or other suitable buffer, the pH may be between 4.6 and 8.8, and the pH may preferably be close to about 7. The lactic acid sensor 100 and the reference electrode 420 can be placed in the solution 405 to be tested. When the enzyme layer in the lactic acid sensor 100 is in contact with the lactic acid in the solution 405 to be tested, the lactic acid may be used to oxidize or reduce the lactic acid by the enzyme layer. For example, lactic acid can be decomposed into pyruvic acid and hydrogen ions. When the above hydrogen ions are generated, the potential in the buffer solution also changes, and is measured by the lactic acid sensing element array. The signal of the signal and reference electrode measured by the lactic acid sensor 100 can be read by the reading circuit 430 and further processed by the computer device 440 for analysis. A voltage type lactic acid sensor manufactured according to an embodiment of the invention may have a sensitivity of 46.874 mV/mM to 62.682 mV/mM, linearity can range from 0.990 to 0.995. In the lactic acid sensor 100 provided by the present invention, since the electron transfer material is added to the enzyme layer, and the patterned wire layer is directly in contact with the enzyme layer and the semiconductor sensing layer, the lactic acid sensor can further be compared with the conventional lactic acid sensor. Effectively reduce the phenomenon of ion interference.

Preparation of lactic acid sensor [Example 1]

A PET substrate is provided to clean impurities on the substrate with acetone, ethanol and deionized water. Next, sputtering was performed at a working power of 10 W at a ratio of nitrogen:oxygen of 40:20 (sccm), and cerium oxide was sputtered onto the PET substrate. Next, a silver wire was prepared by a screen printing machine and baked at 120 ° C for 20 minutes. Next, an epoxy resin release layer was prepared by a screen printing machine and baked at 120 ° C for 60 minutes.

Next, 2 mL of GPTS was titrated on cerium oxide and baked at 120 ° C for 60 minutes. 2 mg of lactate dehydrogenase, 2 mg of nicotinic amine adenine dinucleotide and 0.8 mg of potassium ferricyanide were added to a 5 mM phosphate buffer solution of pH 7 to form an enzyme solution. 2 μl of the above prepared enzyme solution was titrated into the opening above the GPTS in the cerium oxide, and placed in the refrigerator for 1 day to complete the lactic acid sensor.

[Example 2]

The same procedure as in Example 1 was carried out except that 0.8 mg of potassium ferricyanide was not added.

Measurement of lactic acid concentration [Example 3]

The lactic acid sensor, the reference electrode (Ag/AgCl) prepared in Example 1, and the 0.25-4.0 mM lactic acid test solution were placed in the tank to be tested, and the lactic acid sensor and the reference electrode (Ag/AgCl) were not Into the lactic acid test solution, the response voltage was measured with a lactic acid sensor. Each measurement time is about 5 minutes, and the data acquisition card (USB 6008 and PCI-6010 DAQ) is used to input the signal into the computer device for analysis. The sensitivity is 53.962 mV/mM, and the linearity is 0.991 to 0.995. Figure 5 shows.

Interference ion measurement [Embodiment 4]

The lactic acid sensor prepared in Example 1, the reference electrode (Ag/AgCl), and the 1 mM lactic acid test solution containing 1 mM sodium ion were placed in the tank to be tested, and the lactic acid sensor and the reference electrode (Ag/) were used. AgCl) was immersed in the lactic acid test solution, and the response voltage was measured for about 5 minutes.

[Embodiment 5]

The lactic acid sensor prepared in Example 1, the reference electrode (Ag/AgCl), and the 1 mM lactic acid test solution containing 1 mM potassium ion were placed in the tank to be tested, and the lactic acid sensor and the reference electrode (Ag/) were used. AgCl) was immersed in the lactic acid test solution, and the response voltage was measured for about 5 minutes.

[Embodiment 6]

The lactic acid sensor, the reference electrode (Ag/AgCl) prepared in Example 1, and the 1 mM lactic acid test solution containing 1 Mm of calcium ions were placed in the tank to be tested, and the lactic acid sensor and the reference electrode (Ag/) were used. AgCl) was immersed in the lactic acid test solution, and the response voltage was measured for about 5 minutes.

[Embodiment 7]

The lactic acid sensor prepared in Example 2, a reference electrode (Ag/AgCl), and a 1 mM lactic acid test solution containing 1 mM sodium ion were placed in a tank to be tested, and a lactic acid sensor and a reference electrode (Ag/) were used. AgCl) was immersed in the lactic acid test solution, and the response voltage was measured for about 5 minutes.

[Embodiment 8]

The lactic acid sensor prepared in Example 2, a reference electrode (Ag/AgCl), and a 1 mM lactic acid test solution containing 1 mM potassium ion were placed in a tank to be tested, and a lactic acid sensor and a reference electrode (Ag/) were used. AgCl) was immersed in the lactic acid test solution, and the response voltage was measured for about 5 minutes.

[Embodiment 9]

The lactic acid sensor prepared in Example 2, a reference electrode (Ag/AgCl), and a 1 mM lactic acid test solution containing 1 mM calcium ion were placed in a tank to be tested, and a lactic acid sensor and a reference electrode (Ag) were used. /AgCl) immersed in the lactic acid test solution and measures the response voltage for about 5 minutes.

The response voltage and response voltage difference measured in Examples 5 to 9 (the response voltage after adding the interfering ions minus the response voltage when no interfering ions were added) are summarized in Table 1. Further, by using the solution separation method and the formula (1), the above-described response voltage difference can be converted into the voltage selectivity parameter value (K).

In the formula (1), Z _A is the charge amount of the hydrogen ion, and Z _B is the charge amount of the interference ion. Ej-Ei is the response voltage difference, F is the Faraday constant, R is the ideal gas constant, and T is the temperature at the time of measurement. The response voltage difference can be converted into the voltage selectivity parameter value (K). When the voltage selectivity parameter value (K) is smaller, the smaller the influence of the interference ion on the lactic acid sensor, the better the lactic acid sensor's ability to select lactic acid.

It can be seen from Table 1 that when potassium ferricyanide is added to the enzyme layer of the lactic acid sensor (Examples 4-6), the voltage selectivity parameter value is significantly better than that of potassium ferricyanide. Lactic acid sensors (Examples 7-9), and especially for sodium ions. Therefore, it is clear from the above that when an electron transfer substance (for example, potassium ferricyanide) is added to the enzyme layer of the lactic acid sensor, the selectivity of the lactic acid sensor to lactic acid can be greatly improved, and the lactic acid sensor can be reduced. A phenomenon that is disturbed by other interfering ions.

The lactic acid sensor provided by the present invention as described above is simple in structure and manufacturing method, and requires low cost. Therefore, it is possible to achieve the goal of miniaturization and low-cost sensing elements required by the biomedical industry. At the same time, the invention The lactic acid sensor as provided above can reduce the phenomenon of interference by other ions, thereby further improving the selectivity of the lactic acid sensor.

While the invention has been described above in terms of several preferred embodiments, it is not intended to limit the scope of the present invention, and any one of ordinary skill in the art can make any changes without departing from the spirit and scope of the invention. And the scope of the present invention is defined by the scope of the appended claims.

100‧‧‧Lactic acid sensor

102‧‧‧Substrate

104‧‧‧Lactic acid sensing components

106‧‧‧Isolation

108‧‧‧Semiconductor sensing layer

110‧‧‧Enzyme layer

112‧‧‧Fixed layer

120‧‧‧Wire layer

122‧‧‧Electrical array

124‧‧‧Wire area

126‧‧‧Contact area

130‧‧‧ openings

400‧‧‧Measurement system

405‧‧‧Test solution

410‧‧‧Measurement slot

420‧‧‧ reference electrode

430‧‧‧Readout circuit

440‧‧‧Computer equipment

1 and 2 are each a cross-sectional view and an exploded view of a voltage type lactic acid sensor according to an embodiment of the present invention.

3A to 3C are cross-sectional views showing various methods of manufacturing a voltage type lactic acid sensor in accordance with an embodiment of the present invention.

Figure 4 is a schematic illustration of a metrology system including a voltage lactic acid sensor in accordance with an embodiment of the present invention.

Fig. 5 shows the response voltage when various lactic acid concentrations are measured by a voltage type lactic acid sensor according to an embodiment of the present invention.

100‧‧‧Lactic acid sensor

102‧‧‧Substrate

104‧‧‧Lactic acid sensing components

106‧‧‧Isolation

108‧‧‧Semiconductor sensing layer

110‧‧‧Enzyme layer

112‧‧‧Fixed layer

120‧‧‧Wire layer

Claims (18)

A voltage type lactic acid sensor comprises: a substrate; an array of lactic acid sensing elements, comprising a plurality of lactic acid sensing elements arranged on the substrate and isolated from each other by an isolation layer, wherein each lactic acid sensing element comprises: a semiconductor sensing layer on the substrate; and an enzyme layer on the semiconductor sensing layer, wherein the enzyme layer comprises a catalytic enzyme and an electron transporter; and a wire layer comprising an electrode array at the lactic acid sensing In the array of components. The voltage type lactic acid sensor according to claim 1, wherein the electrode array is located between the semiconductor sensing layer of the lactic acid sensing element and the enzyme layer, and is in contact with the enzyme layer. The voltage lactic acid sensor of claim 1, wherein the wire layer further comprises a contact region outside the array of lactic acid sensing elements and electrically connected to the lactic acid sensing elements, wherein The contact area is not covered by the isolation layer. The voltage type lactic acid sensor according to claim 1, wherein the semiconductor sensing layer comprises cerium oxide, tin oxide, aluminum oxide, aluminum nitride, zinc oxide, titanium oxide or a combination thereof. The voltage type lactic acid sensor according to claim 1, wherein the catalytic enzyme comprises lactate oxidase or lactate dehydrogenase. A voltage type lactic acid sensor as described in claim 5, Wherein the enzyme layer further comprises nicotine amidoxime dinucleotide, flavin adenine dinucleotide, flavin mononucleotide or a combination thereof. The voltage type lactic acid sensor according to claim 1, wherein the electron transporting substance comprises potassium ferricyanide, potassium ferrocyanide, tetrathiafulvalene, tetracyanoquinodimethane or Combination of the foregoing. The voltage type lactic acid sensor according to claim 1, wherein the lactic acid enzyme is immobilized on the semiconductor sensing layer by a carrier comprising 3-glycidylpropyltrimethyl Decane, glutaraldehyde, perfluorosulfonic acid or a combination of the foregoing. A method for manufacturing a voltage type lactic acid sensor, comprising: providing a substrate; forming a semiconductor sensing layer on the substrate, wherein the semiconductor sensing layer comprises a plurality of sensing patterns; forming a wire layer on the substrate, The wire layer includes an electrode array at least partially covering the sensing patterns; forming an isolation layer covering the substrate and exposing the sensing patterns; and forming an enzyme layer on the exposed sensing patterns, To form an array of lactic acid sensing elements, wherein the enzyme layer comprises a catalytic enzyme and an electron transporter. The method of manufacturing a voltage type lactic acid sensor according to claim 9, wherein the wire layer further comprises a contact region outside the array of the lactic acid sensing element and electrically connected to the lactic acid sensing elements. Where the contact area is not covered by the isolation layer. A voltage type lactic acid sensor as described in claim 9 The method of producing, wherein the catalytic enzyme comprises lactate oxidase or lactate dehydrogenase. The method for manufacturing a voltage type lactic acid sensor according to claim 11, wherein the enzyme layer further comprises nicotinic amidoxime dinucleotide, flavin adenine dinucleotide, and flavin mononucleoside. Acid or a combination of the foregoing. The method for manufacturing a voltage type lactic acid sensor according to claim 9, wherein the electron transporting substance comprises potassium ferricyanide, potassium ferrocyanide, tetrathiafulvalene, tetracyanoquinone Dimethane or a combination of the foregoing. The method for manufacturing a voltage type lactic acid sensor according to claim 9, wherein the enzyme layer is fixed on the exposed sensing patterns by a carrier, wherein the carrier comprises 3-epoxy-propyl. Alcohol propyl trimethyl decane, glutaraldehyde, perfluoro sulfonic acid or a combination of the foregoing. The method for manufacturing a voltage type lactic acid sensor according to claim 9, wherein the step of forming the enzyme layer on the exposed sensing patterns comprises: adding the catalytic enzyme, the coenzyme, and the electron transfer The solution is formed into a buffer solution to form an enzyme solution; and the enzyme solution is titrated onto the exposed sensing patterns. The method for manufacturing a voltage type lactic acid sensor according to claim 13 further comprises: after titrating the enzyme solution onto the exposed sensing patterns, and storing in an environment below room temperature for 24 hours; . A measuring system comprising a lactic acid sensor, comprising: the voltage type lactic acid sensor according to claim 1; a reference electrode; a measuring tank, providing a solution to be tested, the voltage type lactic acid a sensor and the reference electrode are placed therein; A computer device is electrically connected to the voltage type lactic acid sensor. The measuring system of claim 17, wherein the wire layer of the voltage type lactic acid sensor further comprises a contact area, and the computer device transmits through the contact area The voltage type lactic acid sensor is electrically connected.