An electrochemical detection chip and its preparation method and application

Abstract

The present invention discloses an electrochemical detection chip and a preparation method and application thereof. The electrochemical detection chip of the present invention comprises a working electrode, an auxiliary electrode, a reference electrode and a micro-reaction cell arranged on a chip substrate; the working electrode, the auxiliary electrode and the reference electrode all have extensions in the micro-reaction cell. The electrochemical detection chip of the present invention has the characteristics of strong practicality, high sensitivity, stable performance, short detection time, few detection samples required, simple detection process, etc., and can be used in the fields of food detection, agricultural product quality control, etc. The preparation method of the present invention can realize large-scale and efficient production of the electrochemical detection chip. The electrochemical detection chip of the present invention can be used for rapid detection of hesperidin alone in a solution, rapid detection of naringin alone in a solution, or rapid detection of hesperidin and hesperidin simultaneously in a solution.

Description

Technical Field

[0001] The present invention specifically relates to the technical field of electrochemical chips, and specifically relates to an electrochemical detection chip and a preparation method and application thereof. Background Art

[0002] As people's living standards continue to improve, they have higher requirements for the quality of life. Flavonoids are a class of natural physiologically active substances widely found in agricultural and sideline products such as fruits, vegetables, grains and tea, and have multiple biological activities such as antioxidant, anti-inflammatory, anti-tumor, antibacterial and antiviral. The content of flavonoids often determines the quality of these agricultural and sideline products and their processed products. For example, the "Chinese Pharmacopoeia" stipulates that the content of naringin in Huazhou tangerine peel shall not be less than 3.5%, and the content of hesperidin in tangerine peel shall not be less than 2.0%.

[0003] Traditional flavonoid analysis and detection methods include UV-visible spectrophotometry, high performance liquid chromatography, gas chromatography, etc. These methods generally have problems such as cumbersome sample processing, complex instruments and equipment, time-consuming analysis and detection, and high costs. They are especially not suitable for the rapid determination of flavonoid content in food and agricultural products in daily life.

[0004] In recent years, electrochemical analysis technology has received widespread attention, but there is currently no traditional electrochemical chip suitable for the detection of plant flavonoids including hesperidin, and the detection accuracy is low. At the same time, the electrode preparation method of traditional electrochemical chips is relatively complicated, and the electrode materials are usually expensive, which limits its widespread use in actual production. Summary of the invention

[0005] The purpose of the present invention is to solve the problems of low efficiency of conventional detection of plant flavonoids including hesperidin in the prior art, low electrochemical detection accuracy, high cost, etc. The present invention provides an electrochemical detection chip.

[0006] Another object of the present invention is to provide a method for preparing the electrochemical detection chip described above.

[0007] The present invention also aims to provide an application of the electrochemical detection chip described above, which can be specifically applied to the rapid detection of the concentration of hesperidin, the rapid detection of the concentration of naringin, or the simultaneous rapid detection of the concentrations of hesperidin and naringin.

[0008] The purpose of the present invention is achieved through the following technical solutions.

[0009] An electrochemical detection chip comprises a chip substrate, on which a working electrode, an auxiliary electrode, a reference electrode and a micro-reaction cell are arranged; the working electrode, the auxiliary electrode and the reference electrode all have extensions in the micro-reaction cell;

[0010] The chip substrate is a polyimide film; the working electrode and the auxiliary electrode are produced on the chip substrate by laser induction.

[0011] In a preferred embodiment, the chip substrate has a thickness of 0.5-2.5 mm.

[0012] In a preferred embodiment, the reference electrode is formed by coating and drying a conductive silver paste or a silver-silver chloride paste on the chip substrate.

[0013] In a preferred embodiment, the micro reaction pool is a micro groove formed by gluing a round-hole silica gel sheet on the chip substrate.

[0014] In a preferred embodiment, in any one of the electrochemical detection chips, the micro-reaction cell has a diameter of 1.0 to 1.5 cm and a height of 0.5 to 2.0 mm.

[0015] A method for preparing an electrochemical detection chip comprises the following steps:

[0016] S1. Design the structure of the electrochemical detection chip, including the position distribution of the working electrode, auxiliary electrode, reference electrode and micro-reaction cell;

[0017] S2, laser engraving the working electrode and the auxiliary electrode on the polyimide film;

[0018] S3, uniformly coating the conductive silver paste or silver-silver chloride paste on the designed distribution area of ​​the reference electrode, and baking at 80-120° C. for 1-5 hours to obtain a three-electrode electrochemical chip;

[0019] S4, after uniformly mixing the polydimethylsiloxane prepolymer and the curing agent, drying and curing at 60-100° C. for 10-60 minutes to obtain a silica gel sheet, wherein the thickness of the silica gel sheet is 0.5-2.0 mm; punching holes on the surface of the silica gel sheet, wherein the diameter of the circular holes is 1.0-1.5 cm, to obtain a circular hole silica gel sheet;

[0020] S5, gluing the circular hole silicone sheet to the three-electrode electrochemical chip to obtain the electrochemical detection chip.

[0021] In a preferred embodiment, the laser wavelength of the laser engraving is 405 nm, and the laser power is 1500-3000 mW. Graphene with a three-dimensional structure is generated on the surface of the polyimide film by laser induction, thereby preparing an electrochemical chip.

[0022] The application of the electrochemical detection chip described in any of the above items includes rapid detection of the concentration of hesperidin, rapid detection of the concentration of naringin, or simultaneous rapid detection of the concentrations of hesperidin and naringin. Specifically, hesperidin and naringin undergo an electrocatalytic oxidation reaction on the electrochemical detection chip to generate a catalytic current, and according to the voltage position when the catalytic current is generated, it can be determined that the substance being tested is hesperidin or naringin, and further, the concentration of hesperidin and naringin can be calculated according to the magnitude of the generated catalytic current.

[0023] The use of the electrochemical detection chip described in any one of the above items in the rapid detection of hesperidin concentration, calculating the concentration of hesperidin by the magnitude of the catalytic current generated by the electrochemical reaction of the hesperidin solution on the electrochemical detection chip, comprises the following steps:

[0024] S1. Connecting the electrochemical detection chip to the corresponding interface of the electrochemical workstation;

[0025] S2, adding a buffer solution to the micro-reaction cell of the electrochemical detection chip, and measuring and obtaining a linear voltammetric curve of the blank solution;

[0026] S3, adding hesperidin solutions of different concentrations to the micro-reaction cell of the electrochemical detection chip, and measuring and obtaining voltammetric curves of the hesperidin solutions of different concentrations;

[0027] S4. According to the voltammetric curves corresponding to the hesperidin solutions with different concentrations, the catalytic current value corresponding to each voltammetric curve is determined, and then the relationship between the hesperidin concentration and the catalytic current is determined, and a concentration-current curve model is prepared;

[0028] S5. Add the target hesperidin solution to the micro-reaction cell of the electrochemical detection chip, measure the current value of the target hesperidin solution, and compare it with the concentration-current curve model to determine the concentration of hesperidin in the target hesperidin solution.

[0029] The use of the electrochemical detection chip described in any one of the above items in the rapid detection of naringin concentration, calculating the concentration of naringin by the magnitude of the catalytic current generated by the electrochemical reaction of the naringin solution on the electrochemical detection chip, comprises the following steps:

[0030] S1. Connecting the electrochemical detection chip to the corresponding interface of the electrochemical workstation;

[0031] S2, adding a buffer solution to the micro-reaction cell of the electrochemical detection chip, and measuring and obtaining a linear voltammetric curve of the blank solution;

[0032] S3, adding naringin solutions of different concentrations to the micro-reaction cell of the electrochemical detection chip, and measuring and obtaining voltammetric curves of the naringin solutions of different concentrations;

[0033] S4, according to the voltammetric curves corresponding to the naringin solutions with different concentrations, determining the catalytic current value corresponding to each voltammetric curve, and then determining the relationship between the naringin concentration and the catalytic current, and making a concentration-current curve model;

[0034] S5. Add the target naringin solution to the micro-reaction cell of the electrochemical detection chip, measure the current value of the target naringin solution, and compare it with the concentration-current curve model to determine the concentration of naringin in the target naringin solution.

[0035] The use of the electrochemical detection chip described in any of the above items in the simultaneous rapid detection of the concentrations of hesperidin and naringin comprises the following steps:

[0036] S1. Connecting the electrochemical detection chip to the corresponding interface of the electrochemical workstation;

[0037] S2, adding a buffer solution to the micro-reaction cell of the electrochemical detection chip, and measuring and obtaining a linear voltammetric curve of the blank solution;

[0038] S3, adding hesperidin / naringin mixed solutions of different concentrations to the micro-reaction cell of the electrochemical detection chip, measuring and obtaining voltammetric curves of the hesperidin / naringin mixed solutions of different concentrations;

[0039] S4, according to the voltammetric curves corresponding to the mixed solutions of hesperidin / naringin with different concentrations, determine the catalytic current values ​​of hesperidin and naringin corresponding to each voltammetric curve, and then determine the relationship between hesperidin, naringin and the catalytic current, and make a concentration-current curve model;

[0040] S5. Add the target hesperidin / naringin mixed solution to the micro-reaction cell of the electrochemical detection chip, measure the current value of the target hesperidin / naringin mixed solution, and compare it with the concentration-current curve model to determine the concentrations of hesperidin and naringin in the hesperidin / naringin mixed solution.

[0041] In a preferred embodiment, in any of the above applications, the amount of the buffer solution added is 100 μL to 200 μL.

[0042] In a preferred embodiment, in any of the above applications, the voltammetric curve is measured and obtained under the conditions of a potential range of 0.7 to 0.2 V and a scan rate of 0.01 V / s. The voltammetric curve includes a cyclic voltammetric curve or a linear voltammetric scan curve.

[0043] In a preferred embodiment, in any of the above uses, the buffer solution comprises a phosphate buffer solution.

[0044] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0045] The electrochemical detection chip of the present invention has high integration, small size, simple structure, low cost, strong practicality, high sensitivity (the detection sensitivity of hesperidin and naringin can reach 13.8μA / mmol / L and 18.8μA / mmol / L respectively), short detection time (single detection time 50s), low detection limit (the minimum detection concentration of hesperidin and naringin can reach 5×10 -8 mmol / L), high spike recovery rate (90% to 110%), small amount of test sample required, simple test process, and easy preparation. It can be used in food testing, agricultural product quality control and other fields, especially for the rapid evaluation of food and drug quality and agricultural product quality in daily life, and has broad application prospects.

[0046] The preparation method of the present invention can realize large-scale and efficient production of the electrochemical detection chip, thereby facilitating the wide application of the electrochemical detection chip.

[0047] The electrochemical detection chip of the present invention has a highly sensitive response to hesperidin and naringin in a solution, can realize rapid detection of hesperidin alone in a solution, rapid detection of naringin alone in a solution, or simultaneous rapid detection of hesperidin and hesperidin in a solution, and can be used for real-time and rapid detection of the content of hesperidin and naringin in an actual sample. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 It is a structural schematic diagram of the electrochemical detection chip of the present invention;

[0049] Figure 2 This is a scanning electron microscope image of the working electrode of the electrochemical detection chip of Example 1;

[0050] Figure 3 This is a linear voltammetric curve diagram of detecting hesperidin in Example 2;

[0051] Figure 4 This is a model diagram of the concentration-current curve for detecting hesperidin in Example 2;

[0052] Figure 5 This is a detection curve diagram of hesperidin in Example 2;

[0053] Figure 6 This is a linear voltammetric curve diagram for detecting naringin in Example 3;

[0054] Figure 7This is a model diagram of the concentration-current curve for detecting naringin in Example 3;

[0055] Figure 8 This is a detection curve diagram of naringin in Example 3;

[0056] Fig. 9 This is a linear voltammetric curve diagram of the simultaneous detection of hesperidin and naringin in Example 4;

[0057] Fig.10 This is a concentration-current curve model diagram for simultaneous detection of hesperidin and naringin in Example 4;

[0058] Fig.11 This is a detection curve diagram of the simultaneous detection of a mixed solution of hesperidin and naringin in Example 4;

[0059] Figure markings: 1-chip substrate, 2-working electrode, 3-auxiliary electrode, 4-reference electrode, 5-micro reaction cell. DETAILED DESCRIPTION

[0060] The technical solution of the present invention is further described in detail below in conjunction with specific embodiments, but the protection scope and implementation mode of the present invention are not limited thereto. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0061] Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without any creative work.

[0062] Unless otherwise defined, all technical and scientific terms used herein are identical with the meanings generally understood by the technicians belonging to the technical field of the present invention, and the reagents, equipment and methods used in the present invention are conventional commercially available reagents, equipment and conventional methods used in the technical field. The terms used in the specification of the present invention herein are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0063] Unless otherwise specified, all technical and scientific terms used herein have the standard meaning in the field to which the claimed subject matter belongs. If there are multiple definitions for a term, the definition herein shall prevail.

[0064] Unless otherwise indicated, the present invention employs standard nomenclature and standard laboratory procedures and techniques of analytical chemistry, synthetic organic chemistry, and optics. In some cases, standard techniques are used for chemical syntheses and chemical analyses.

[0065] It should be understood that the singular forms used in the present invention, such as "a", include plural references unless otherwise specified. In addition, the terms "including", "containing", and "having" are open-ended definitions rather than closed-ended definitions, that is, they include the contents specified in the present invention, but do not exclude other aspects. In other words, the terms also include "essentially consisting of" or "consisting of". In addition, "and combinations thereof" in the specification refers to any combination of all the items listed.

[0066] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0067] The electrochemical detection chip of the present invention, please refer to Figure 1 As shown, it comprises a chip substrate 1. A working electrode 2, an auxiliary electrode 3, a reference electrode 4 and a micro reaction cell 5 are arranged on the chip substrate 1.

[0068] Wherein, the chip substrate 1 is a polyimide film, and the thickness of the chip substrate 1 is 0.5-2.5 mm. In some specific embodiments, the length of the chip substrate 1 is 2-5 cm, and the width is 1-2 cm. The working electrode 2 and the auxiliary electrode 3 are produced by laser induction on the chip substrate 1, and the reference electrode 4 is formed by coating and drying the conductive silver paste or silver chloride paste on the chip substrate 1. The micro-reaction pool 5 is a micro-groove formed by gluing a circular hole silica gel sheet on the chip substrate 1, and the diameter of the micro-reaction pool 5 is 1-1.5 cm, and the height is 0.5-2.0 mm.

[0069] Moreover, the working electrode 2, the auxiliary electrode 3 and the reference electrode 4 are all extended in the micro-reaction cell 4. During the detection application, the solution to be detected reacts in the micro-reaction cell 5, and the generated catalytic current causes the working electrode 2 to have a potential difference relative to the reference electrode 4, and the auxiliary electrode 3 is polarized, so that the voltage position of the catalytic current can be detected, and then the concentration of the solution to be detected can be calculated.

[0070] Embodiment 1

[0071] The preparation of the electrochemical detection chip of this embodiment is carried out in the following specific steps:

[0072] S1. Use drawing software to design an electrochemical chip on a computer, including the position distribution of the working electrode, auxiliary electrode, reference electrode and micro-reaction cell.

[0073] S2. Print the working electrode and the auxiliary electrode on the polyimide film by using a laser engraving machine; wherein the laser wavelength is 405 nm and the laser power is 1800 mW.

[0074] S3. Evenly apply the conductive silver paste on the reference electrode area, and then put it into a drying oven. The temperature is controlled at 90 degrees Celsius and the drying time is controlled at 4 hours to obtain a three-electrode electrochemical chip.

[0075] S4. Mix the polydimethylsiloxane prepolymer (such as Dow Corning SYLGARD184) and the curing agent (such as Dow Corning SYLGUARD-184B) in a mass ratio of 10:1, pour into a culture dish, wait for it to be leveled, and then put it into a drying oven for curing. The curing temperature is controlled at 60 degrees Celsius, the time is controlled at 30 minutes, and the thickness is 1 mm.

[0076] A hole puncher was used to punch holes on the surface of polydimethylsiloxane to form a round-hole polydimethylsiloxane sheet, and the hole diameter was controlled to be 1 cm.

[0077] S5. Adhere the circular hole polydimethylsiloxane sheet to the three-electrode electrochemical chip to obtain the electrochemical detection chip.

[0078] The schematic diagram of the structure of the prepared electrochemical detection chip is shown in Figure 1 As shown, and the scanning electron microscope image of the working electrode is as shown Figure 2 As shown by Figure 2 It can be seen that the working electrode has a three-dimensional porous surface structure, and therefore has a large specific surface area and high detection sensitivity.

[0079] Embodiment 2

[0080] The electrochemical detection chip of Example 1 is used to detect the concentration of hesperidin, and the steps are as follows:

[0081] S1. Connect the electrochemical detection chip to the corresponding interface of the electrochemical workstation;

[0082] S2. Add 200 μL of phosphate buffer solution to the micro-reaction cell of the electrochemical detection chip, and measure the linear voltammetric curve of the blank solution under the conditions of a potential scanning range of 0.7 to 0.2 V and a scanning rate of 0.01 V / s;

[0083] S3. Different amounts of hesperidin solution were added to the phosphate buffer solution to make the final concentrations of 0.00005mmol / L, 0.0001mmol / L, 0.0005mmol / L, 0.001mmol / L, 0.002mmol / L, 0.004mmol / L, 0.006mmol / L, 0.008mmol / L, 0.01mmol / L, 0.015mmol / L, 0.020mmol / L, 0.030mmol / L, 0.040mmol / L, 0.060mmol / L, 0.080mmol / L and 0.100mmol / L. Linear voltammetry was performed at a potential scanning range of 0.7 to 0.2V and a scanning rate of 0.01V / s to obtain a linear voltammetry curve, as shown in FIG. Figure 3 As shown; among them, Figure 3 The results show that as the concentration of hesperidin added increases, the catalytic current obtained increases accordingly.

[0084] S4. By subtracting the background current from the linear voltammetric curve by tangent, the peak current value of the linear voltammetric curve at different concentrations is read, which is the catalytic current value. The corresponding relationship between the peak current value and the hesperidin concentration is established to obtain the concentration-current curve model and perform linear fitting; Figure 4 As shown, the linear relationship of the concentration-current curve model is: y = 13.803x + 0.1403 (R 2 =0.96). The results show that the obtained electrochemical detection chip can achieve linear detection of hesperidin in the range of 0.00005mmol / L to 0.100mmol / L.

[0085] S5. Add hesperidin solution to the micro-reaction cell of the electrochemical detection chip to a target concentration of 0.025 mmol / L to obtain a target hesperidin solution; the peak current value of the target hesperidin solution is measured to be 0.503 μA, and the corresponding detection curve is as follows: Figure 5 As shown, compared with the concentration-current curve model, the calculated hesperidin concentration was 0.026 mmol / L and the recovery rate was 104%.

[0086] Embodiment 3

[0087] The electrochemical detection chip of Example 1 is used to detect the concentration of naringin, and the steps are as follows:

[0088] S1. Connect the electrochemical detection chip to the corresponding interface of the electrochemical workstation;

[0089] S2. Add 200 μL of phosphate buffer solution to the micro-reaction cell of the electrochemical detection chip, and measure the linear voltammetric curve of the blank solution under the conditions of a potential scanning range of 0.7 to 0.2 V and a scanning rate of 0.01 V / s;

[0090] S3. Add different amounts of naringin solution to the phosphate buffer solution, and make the final concentrations of 0.00005mmol / L, 0.0001mmol / L, 0.0005mmol / L, 0.001mmol / L, 0.002mmol / L, 0.004mmol / L, 0.006mmol / L, 0.008mmol / L, 0.01mmol / L, 0.015mmol / L, 0.020mmol / L, 0.030mmol / L, 0.040mmol / L, 0.060mmol / L, 0.080mmol / L, 0.100mmol / L, and perform linear voltammetry scanning in the potential scanning range of 0.7-0.2V and the scanning rate of 0.01V / s to obtain a linear voltammetry curve, as shown in FIG. Figure 6 As shown; among them, Figure 6 The results show that as the concentration of hesperidin added increases, the catalytic current obtained increases accordingly.

[0091] S4. By subtracting the background current from the linear voltammetric curve by tangent, the peak current value of the linear voltammetric curve at different concentrations is read, which is the catalytic current value. The corresponding relationship between the peak current value and the concentration of naringin is established to obtain the concentration-current curve model and perform linear fitting; Figure 7 As shown, the linear relationship of the concentration-current curve model is: y = 18.775x + 0.1882 (R 2 =0.97). The results show that the obtained electrochemical monitoring chip can achieve linear detection of naringin in the range of 0.00005mmol / L to 0.100mmol / L.

[0092] S5. Add naringin solution to the micro-reaction cell of the electrochemical detection chip to make its target concentration 0.025 mmol / L, and obtain the target naringin solution; the peak current value of the target naringin solution is measured to be 0.678 μA, and the corresponding detection curve is as follows: Figure 8 As shown, compared with the concentration-current curve model, the calculated naringin concentration was 0.026 mmol / L, and the recovery rate was 104%.

[0093] Embodiment 4

[0094] The electrochemical detection chip of Example 1 is used to synchronously and rapidly detect the concentrations of hesperidin and naringin, and the steps are as follows:

[0095] S1. Connect the electrochemical detection chip to the corresponding interface of the electrochemical workstation;

[0096] S2. Add 200 μL of phosphate buffer solution to the micro-reaction cell of the electrochemical detection chip, and measure the linear voltammetric curve of the blank solution under the conditions of a potential scanning range of 0.7 to 0.2 V and a scanning rate of 0.01 V / s;

[0097] S3. Different amounts of hesperidin / naringin mixed solution were added to the phosphate buffer solution to make the final concentrations of 0.00005mmol / L, 0.0001mmol / L, 0.0005mmol / L, 0.001mmol / L, 0.002mmol / L, 0.004mmol / L, 0.006mmol / L, 0.008mmol / L, 0.01mmol / L, 0.015mmol / L, 0.020mmol / L, 0.030mmol / L, 0.040mmol / L, 0.060mmol / L, 0.080mmol / L and 0.100mmol / L. Linear voltammetry was performed under the conditions of a potential scanning range of 0.7 to 0.2V and a scanning rate of 0.01V / s to obtain a linear voltammetry curve, as shown in FIG. Fig. 9 As shown; among them, Fig. 9 The results show that as the concentration of hesperidin and naringin added increases, the catalytic current obtained increases accordingly.

[0098] S4. By subtracting the background current from the linear voltammetric curve by tangent, the peak current value of the linear voltammetric curve at different concentrations is read, which is the catalytic current value. The corresponding relationship between the peak current value and the concentration of hesperidin and naringin is established to obtain a concentration-current curve model and perform linear fitting; Fig.10 As shown, the linear relationship of the concentration-current curve model is: y = 13.901x + 0.112 (R 2 = 0.98, hesperidin) and y = 18.076x + 0.1231 (R 2 =0.98, naringin). The results show that the obtained electrochemical chip can achieve linear detection in the range of 0.00005mmol / L to 0.100mmol / L in the mixed solution of hesperidin and naringin.

[0099] S5. Add a mixed solution of hesperidin and naringin to the micro-reaction cell of the electrochemical detection chip, so that the target concentrations of both are 0.025 mmol / L, to obtain a target hesperidin / naringin mixed solution; the peak current values ​​of hesperidin and naringin corresponding to the target hesperidin / naringin mixed solution are measured to be 0.484 μA and 0.544 μA, respectively. The corresponding detection curve is as follows: Fig.11As shown, compared with the concentration-current curve model, the calculated concentrations of hesperidin and naringin were 0.027 mmol / L and 0.023 mmol / L, respectively, and the recovery rates were 108% and 92%, respectively.

[0100] The technical features of the above-mentioned embodiments can be combined arbitrarily. In order to make the description concise, this specification does not describe all possible combinations of the technical features in the above-mentioned embodiments. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification. Moreover, the above-mentioned embodiments only express several implementation methods of the present invention, and the description is relatively specific and detailed, but it cannot be understood as a limitation on the scope of the invention patent.

[0101] It should be noted that, for those skilled in the art, several modifications and improvements can be made without departing from the concept of the present invention, and these modifications and improvements all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. An electrochemical detection chip for detecting the concentration of hesperidin and / or naringin, comprising a chip substrate, characterized in that: The chip substrate is provided with a working electrode, an auxiliary electrode, a reference electrode and a micro-reaction cell; the working electrode, the auxiliary electrode and the reference electrode all have extensions in the micro-reaction cell; The chip substrate is a polyimide film; the working electrode and the auxiliary electrode are produced on the chip substrate by laser induction; The reference electrode is formed by coating and drying a conductive silver paste or a silver-silver chloride paste on the chip substrate; The solution to be tested reacts in the micro-reaction cell, and the generated catalytic current causes the working electrode to have a potential difference relative to the reference electrode, and the auxiliary electrode is polarized, so that the voltage position of the catalytic current can be detected, and then the concentration of the solution to be tested can be calculated.

2. The electrochemical detection chip according to claim 1, characterized in that: The micro reaction pool is a micro groove formed by gluing a round-hole silica gel sheet on the chip substrate.

3. The electrochemical detection chip according to any one of claims 1 to 2, characterized in that: The diameter of the micro reaction pool is 1.0-1.5 cm, and the height is 0.5-2.0 mm.

4. Use of the electrochemical detection chip according to any one of claims 1 to 3 in rapid detection of hesperidin concentration, characterized in that: The steps include: S1. Connecting the electrochemical detection chip to the corresponding interface of the electrochemical workstation; S2, adding a buffer solution to the micro-reaction cell of the electrochemical detection chip, and measuring and obtaining a linear voltammetric curve of the blank solution; S3, adding hesperidin solutions of different concentrations to the micro-reaction cell of the electrochemical detection chip, measuring and obtaining voltammetric curves of the hesperidin solutions of different concentrations; S4. According to the voltammetric curves corresponding to the hesperidin solutions with different concentrations, the catalytic current value corresponding to each voltammetric curve is determined, and then the relationship between the hesperidin concentration and the catalytic current is determined, and a concentration-current curve model is prepared; S5. Add the target hesperidin solution to the micro-reaction cell of the electrochemical detection chip, measure the current value of the target hesperidin solution, and compare it with the concentration-current curve model to determine the concentration of hesperidin in the target hesperidin solution.

5. Use of the electrochemical detection chip according to any one of claims 1 to 3 in rapid detection of naringin concentration, characterized in that: The steps include: S1. Connecting the electrochemical detection chip to the corresponding interface of the electrochemical workstation; S2, adding a buffer solution to the micro-reaction cell of the electrochemical detection chip, and measuring and obtaining a linear voltammetric curve of the blank solution; S3, adding naringin solutions of different concentrations to the micro-reaction cell of the electrochemical detection chip, and measuring and obtaining voltammetric curves of the naringin solutions of different concentrations; S4, according to the voltammetric curves corresponding to the naringin solutions with different concentrations, determining the catalytic current value corresponding to each voltammetric curve, and then determining the relationship between the naringin concentration and the catalytic current, and making a concentration-current curve model; S5. Add the target naringin solution to the micro-reaction cell of the electrochemical detection chip, measure the current value of the target naringin solution, and compare it with the concentration-current curve model to determine the concentration of naringin in the target naringin solution.

6. Use of the electrochemical detection chip according to any one of claims 1 to 3 in the simultaneous and rapid detection of hesperidin and naringin concentrations, characterized in that: The steps include: S1. Connecting the electrochemical detection chip to the corresponding interface of the electrochemical workstation; S2, adding a buffer solution to the micro-reaction cell of the electrochemical detection chip, and measuring and obtaining a linear voltammetric curve of the blank solution; S3, adding hesperidin / naringin mixed solutions of different concentrations to the micro-reaction cell of the electrochemical detection chip, measuring and obtaining voltammetric curves of the hesperidin / naringin mixed solutions of different concentrations; S4, according to the voltammetric curves corresponding to the mixed solutions of hesperidin / naringin with different concentrations, determine the catalytic current values ​​of hesperidin and naringin corresponding to each voltammetric curve, and then determine the relationship between hesperidin, naringin and the catalytic current, and make a concentration-current curve model; S5. Add the target hesperidin / naringin mixed solution to the micro-reaction cell of the electrochemical detection chip, measure the current value of the target hesperidin / naringin mixed solution, and compare it with the concentration-current curve model to determine the concentrations of hesperidin and naringin in the hesperidin / naringin mixed solution.

7. The use according to any one of claims 4, 5 and 6, characterized in that: The voltammetric curve was measured under the conditions of a potential scanning range of 0.7-0.2 V and a scanning rate of 0.01 V / s.

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