WO2023179806A2 - Biochemical substance measurement system and method, and storage medium - Google Patents

Biochemical substance measurement system and method, and storage medium Download PDF

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Publication number
WO2023179806A2
WO2023179806A2 PCT/CN2023/106207 CN2023106207W WO2023179806A2 WO 2023179806 A2 WO2023179806 A2 WO 2023179806A2 CN 2023106207 W CN2023106207 W CN 2023106207W WO 2023179806 A2 WO2023179806 A2 WO 2023179806A2
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WIPO (PCT)
Prior art keywords
electrode
working electrode
electrode group
biochemical substance
extraction
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PCT/CN2023/106207
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French (fr)
Chinese (zh)
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WO2023179806A3 (en
Inventor
杨玲
陈毅豪
杨芷炎
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杭州柔谷科技有限公司
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Priority to PCT/CN2023/106207 priority Critical patent/WO2023179806A2/en
Publication of WO2023179806A2 publication Critical patent/WO2023179806A2/en
Publication of WO2023179806A3 publication Critical patent/WO2023179806A3/en

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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00Measuring for diagnostic purposes; Identification of persons
    • A61B5/145Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue

Definitions

  • This application relates to the technical field of biochemical substance detection, and specifically relates to a biochemical substance detection system, method and storage medium.
  • the normal life activities of the human body are inseparable from the stimulation and regulation of various biochemical substances in the body. Changes in one or several biochemical indicators will affect the normal operation of the human body and even affect life and health.
  • the human body's blood glucose concentration levels and fluctuations are important monitoring and disease management indicators for patients with abnormal glucose metabolism; luteinizing hormone is one of the important indicators for sex hormone detection, and its concentration level can be used to guide diagnosis and efficacy evaluation.
  • this application provides a biochemical substance detection system, method and storage medium.
  • the detection area is electroporated first and then the biochemical substances are extracted. This can improve the transdermal extraction efficiency of biochemical substances and is more suitable for macromolecular substances. extraction and detection.
  • the present application provides a biochemical substance detection system, including a first electrode group, a second electrode group and a processing device, and the processing device is connected to the first electrode group and the second electrode group;
  • the first electrode group includes a first working electrode with a first biosensing layer modified on its surface
  • the second electrode group includes a second working electrode with a second biosensing layer modified on its surface.
  • the sensing layer and the second biosensing layer are used to detect the same or different biochemical substances;
  • the processing equipment is configured for:
  • the biochemical substance is detected through the first working electrode and/or the second working electrode to determine the concentration of the biochemical substance.
  • the first electrode group further includes a first pair of electrodes, a first reference electrode, a first extraction electrode and a first flexible substrate, and the first working electrode, the first pair of electrodes, the first reference electrode The specific electrode and the first extraction electrode are arranged on the first flexible substrate;
  • the second electrode group also includes a second counter electrode, a second reference electrode, a second extraction electrode and a second flexible substrate.
  • the second working electrode, the second counter electrode, the second reference electrode and the second extraction electrode Electrodes are provided on the second flexible substrate;
  • the first extraction electrode and the second extraction electrode are used to apply the pulse voltage and the constant current to the detection area.
  • the first working electrode is a sheet electrode
  • the first pair of electrodes and the first reference electrode are arranged around the periphery of the first working electrode
  • the first extraction electrode surrounds The peripheral arrangement of the first pair of electrodes
  • the second working electrode is a sheet electrode
  • the second pair of electrodes and the second reference electrode surround the outer periphery of the second working electrode
  • the second extraction electrode surrounds the outer periphery of the second pair of electrodes.
  • the first electrode group is provided with a first support ring, and the first support ring is arranged around the periphery of the electrodes in the first electrode group;
  • the second electrode group is provided with a second support ring, and the second support ring is arranged around the periphery of the electrodes in the second electrode group.
  • the processing device when the biochemical substance is detected through the first working electrode and/or the second working electrode, and the concentration of the biochemical substance is determined, the processing device is configured to:
  • Differential pulse voltammetry is used to determine the concentration of the biochemical substance based on the electrochemical signals generated by the first working electrode and/or the second working electrode.
  • the pulse parameters for applying the pulse voltage include pulse voltage, pulse width, period and number of pulses, the range of the pulse voltage is 100V -500V, and the range of the pulse width is 10 ms - 50 ms, the period ranges from 0 s to 20 s, and the number of pulses ranges from 20 to 200.
  • the current parameters used to apply the constant current include a current value and an extraction time.
  • the current value ranges from 200 ⁇ A to 500 ⁇ A, and the extraction time ranges from 1 min to 10 min.
  • the first biosensing layer and the second biosensing layer are used to detect different biochemical substances, and the biochemical substances include luteinizing hormone and follicle-stimulating hormone.
  • This application also provides a biochemical substance detection method, which is applied to the biochemical substance detection system as described above.
  • the method includes:
  • the biochemical substance is detected through the first working electrode and/or the second working electrode to determine the concentration of the biochemical substance.
  • the present application also provides a computer-readable storage medium.
  • a computer program is stored on the computer-readable storage medium.
  • the computer program is executed by a processor, the steps of the biochemical substance detection method as described above are implemented.
  • the system includes a first electrode group, a second electrode group and processing equipment; the first electrode group includes a first working electrode with a first biosensing layer modified on its surface, and a second The electrode group includes a second working electrode with a second biosensing layer modified on its surface; the processing device is configured to: apply a pulse voltage to the detection area through the first electrode group and the second electrode group to perform electroporation processing; An electrode group and a second electrode group apply a constant current to the detection area to extract biochemical substances into the electrolyte in the area where the first working electrode and the second working electrode are located; the biochemical substances are extracted through the first working electrode and/or the second working electrode.
  • Tests are performed to determine the concentration of biochemical substances.
  • the technical solution of this application first electroporates the detection area and then extracts biochemical substances, which can improve the transdermal extraction efficiency of biochemical substances and is more suitable for the extraction and detection of macromolecular substances.
  • Figure 1 is a schematic structural diagram of a biochemical substance detection system according to the first embodiment.
  • Figure 2 is a schematic diagram of the electroporation channel.
  • Figure 3 is a schematic diagram of an experimental device for biochemical substance detection according to the first embodiment.
  • FIG. 4 is a cross-sectional view of the experimental equipment shown in FIG. 3 .
  • Figure 5 is the DPV response curve of the extraction solution using 150V pulse voltage and different extraction times.
  • Figure 6 is the DPV response curve of the extraction solution using 300V pulse voltage and different extraction times.
  • Figure 7 is a schematic diagram of the extraction results of different processing methods.
  • Figure 8 is a schematic flow chart of a biochemical substance detection method according to the second embodiment.
  • A, B or C or "A, B and/or C” means "any of the following: A; B; C; A and B; A and C; B and C; A, B and C” . Exceptions to this definition occur only when the combination of elements, functions, steps, or operations is inherently mutually exclusive in some manner.
  • FIG. 1 is a schematic structural diagram of a biochemical substance detection system according to the first embodiment.
  • the biochemical substance detection system of this embodiment includes a first electrode group 10 , a second electrode group 20 and a processing device 30 .
  • the processing device 30 is connected to the first electrode group 10 and the second electrode group 20 .
  • the first electrode group 10 includes a first working electrode 11 with a first biosensing layer modified on its surface.
  • the second electrode group 20 includes a second working electrode 21 with a second biosensing layer modified on its surface. The first biosensing layer and the second biosensing layer for detecting the same or different biochemical substances.
  • the processing device 30 is configured to: apply a pulse voltage to the detection area through the first electrode group 10 and the second electrode group 20 to perform electroporation treatment on the detection area; A constant current is applied to the detection area so that the biochemical substances are extracted into the electrolyte in the area where the first working electrode 11 and the second working electrode 21 are located; the biochemical substances are detected through the first working electrode 11 and/or the second working electrode 21, Determine the concentration of biochemical substances.
  • the processing equipment 30 is a large or small electrochemical workstation and can provide the pulse voltage required for electroporation.
  • the first electrode group 10 and the second electrode group 20 can be connected to the processing equipment 30 in a plug-in manner.
  • the first working electrode 11 and the second working electrode 21 can be used to apply pulse voltage and constant current to the detection area and detect biochemical substances to simplify the structure of the electrode group.
  • applying pulse voltage and constant current to the detection area can be performed through other electrodes in the first electrode group 10 and the second electrode group 20 to increase the service life of the first working electrode 11 and the second working electrode 21 .
  • the same or different biochemical substances can be detected by modifying the same or different biosensing layers on the first working electrode 11 and the second working electrode 21 .
  • the first biosensing layer and the second biosensing layer are used to detect different macromolecular biochemical substances, and the macromolecular biochemical substances include luteinizing hormone and follicle-stimulating hormone.
  • the macromolecular biochemical substances include luteinizing hormone and follicle-stimulating hormone.
  • luteinizing hormone as an example, by modifying the surface of the first working electrode 11 with a graphene oxide/thionine/gold nanocomposite, luteinizing hormone antibody, and bovine serum albumin as the first biosensing layer, it can be achieved Testing for luteinizing hormone.
  • a constant current is applied to the detection area through the first electrode group 10 and the second electrode group 20, that is, reverse electroosmosis extraction is performed.
  • an electric field is applied to the skin surface to cause Cl - and Na + in the subcutaneous tissue to move toward the positive and negative electrodes respectively under the action of the electric field, forming a DC current channel from the skin surface through the subcutaneous tissue and back to the skin surface.
  • biochemical substances in subcutaneous tissue fluid can be carried to the skin surface for detection.
  • the extraction efficiency of this method is limited, and it is difficult for macromolecular substances to pass through reverse electroosmosis due to their large diameter. extracted to the skin surface.
  • this application before performing reverse electroosmosis extraction, this application first uses appropriate pulse parameters to electroporate the detection area, thereby improving the transdermal extraction efficiency of biochemical substances and making it more suitable for the extraction and detection of macromolecular substances.
  • a dominant voltage gradient across the skin is established by passing through the non-conductive stratum corneum. If the voltage gradient exceeds the barrier breakdown potential, holes will be formed, allowing substances to penetrate Transmitted through the skin barrier, for example, from the subcutaneous to the skin surface through channels such as hair follicles A, keratinocyte gaps B, sweat glands C, and other pores D. Depending on the pulse parameters used, these channels will re-close after a period of time or return to their original state. After forming a reversible channel on the skin surface and then performing reverse electroosmosis extraction, more target substances can be extracted from tissue fluid and blood, effectively increasing the extraction amount of subcutaneous target substances. At the same time, it is also conducive to improving extraction efficiency and extracting large amounts of target substances. Molecular substances, realizing the detection of macromolecular substances.
  • the first electrode group 10 further includes a first pair of electrodes 12 , a first reference electrode 13 , a first extraction electrode 14 and a first flexible substrate (not shown in FIG. 1 ).
  • the first working electrode 11 , the first pair of electrodes 12, the first reference electrode 13 and the first extraction electrode 14 are arranged on the first flexible substrate.
  • the second electrode group 20 also includes a second counter electrode 22 , a second reference electrode 23 , a second extraction electrode 14 and a second flexible substrate (not shown in FIG. 1 ).
  • the second working electrode 21 and the second counter electrode 22 , the second reference electrode 23 and the second extraction electrode 14 are arranged on the second flexible substrate. Wires and interfaces are provided on the first flexible substrate and the second flexible substrate. Each electrode is connected to the interface through a wire.
  • the interface is used to connect the processing device 30.
  • the arrangement of the wires on the flexible substrate includes a zigzag arrangement and a straight line arrangement. , curve arrangement and other arrangement methods.
  • the materials of the first working electrode 11 and the second working electrode 21 are Pt/C, and the materials of the remaining electrodes can be Ag/AgCl.
  • the flexible base can completely fit the body surface and meet the needs of wearable and easy-to-carry testing.
  • the first working electrode 11 is a sheet electrode
  • the first pair of electrodes 12 and the first reference electrode 13 are arranged around the periphery of the first working electrode 11
  • the first extraction electrode 14 surrounds the periphery of the first pair of electrodes 12 Peripheral settings.
  • the part of the first working electrode 11 located in the detection area is in the shape of a disk, and the first counter electrode 12 and the first reference electrode 13 are commonly surrounding the outer periphery of the disk-shaped part of the first working electrode 11 , that is, surrounding In a circumferential area around the disk-shaped portion of the first working electrode 11, a first pair of electrodes 12 is provided in some areas, and a first reference electrode 13 is provided in another area, thereby reducing the size of the electrodes.
  • the first extraction electrode 14 surrounds the outer periphery of the first pair of electrodes 12 , that is, is disposed on the outermost side of other electrodes in the first electrode group 10 .
  • the second working electrode 21 is a sheet electrode
  • the second counter electrode 22 and the second reference electrode 23 surround the outer periphery of the second working electrode 21
  • the second extraction electrode 14 surrounds the outer periphery of the second counter electrode 22 .
  • the electrodes in the first electrode group 10 and the second electrode group 20 are arranged in the same manner, which will not be described again.
  • the counter electrode is used to form a polarization loop with the corresponding working electrode so that current can flow through the working electrode.
  • the reference electrode is used to provide and maintain a fixed reference potential during the detection of biochemical substances.
  • the first extraction electrode 14 and the second extraction electrode 14 are used to apply pulse voltage to the detection area to perform electroporation treatment, and are also used to apply constant current to the detection area to perform reverse electroosmosis extraction.
  • the pulse parameters for applying the pulse voltage include pulse voltage, pulse width, period and number of pulses.
  • the pulse voltage ranges from 100V to 500V
  • the pulse width ranges from 10 ms to 50 ms.
  • the period ranges from 0 s to 20 s
  • the number of pulses ranges from 20 to 200.
  • the current parameters for applying a constant current include a current value and an extraction time. The current value ranges from 200 ⁇ A to 500 ⁇ A.
  • the extraction time ranges from 1 min to 10 min.
  • the extraction efficiency is highest when the pulse voltage is 150 V or 300 V
  • the number of pulses is 100
  • the constant current is 300 ⁇ A
  • the extraction time is 10 minutes.
  • the first electrode group 10 is provided with a first support ring (not shown in FIG. 1 ), which is disposed around the periphery of the electrodes in the first electrode group 10 , that is, located at the first extraction electrode 14 of the periphery.
  • the second electrode group 20 is provided with a second support ring (not shown in FIG. 1 ).
  • the second support ring is arranged around the periphery of the electrodes in the second electrode group 20 , that is, located on the periphery of the second extraction electrode 14 .
  • the first support ring and the second support ring may be annular adhesive tapes with a certain thickness. One side of the adhesive tape is attached to the flexible base, and the other side is used to adhere to the body surface.
  • a storage space for storing electrolyte can be formed between the electrodes and the body surface through the support ring.
  • An electrical path is formed between the electrode and the body surface.
  • the extracted biochemical substances diffuse in the electrolyte to be in the area of the working electrode to achieve detection.
  • the processing device 30 when detecting biochemical substances through the first working electrode 11 and/or the second working electrode 21 and determining the concentration of the biochemical substances, the processing device 30 is configured to:
  • Differential pulse voltammetry is used to determine the concentration of biochemical substances based on the electrochemical signals generated by the first working electrode 11 and/or the second working electrode 21 .
  • differential pulse voltammetry is used to collect electrochemical signals of the first working electrode 11 and the second working electrode 21 through the processing device 30.
  • the changes of the electrochemical signals of the first working electrode 11 and the second working electrode 21 are related to those to be measured.
  • the concentration of biochemical substances in the liquid is proportional. Therefore, the processing device 30 can determine the concentration of the biochemical substances in the liquid to be tested based on changes in the electrochemical signals of the first working electrode 11 and the second working electrode 21 .
  • this application first electroporates the detection area and then extracts biochemical substances, which can improve the transdermal extraction efficiency of biochemical substances and is more suitable for the extraction and detection of macromolecular substances.
  • the scheme of this application is explained below through experimental data.
  • An experimental equipment includes an electrode group 40, a circuit board 41, a resistance contact 42, a base 43, rat skin 45, a sponge 47, an inner container 46, and an outer container 44.
  • the electrode group 40 includes a first electrode group 10 and a second electrode group 20 .
  • the circuit board 41 is connected to the electrode group and is provided with resistance contacts 42 .
  • the resistance contacts 42 are used to connect to the processing equipment 30 , and the base 43 is used to connect to the processing equipment 30 .
  • the circuit board 41 is supported, and the inner container 46 is used to hold a solution for replacing tissue fluid.
  • a sponge 47 is placed in the inner container 46, and then the rat skin 45 used to simulate the body surface is covered at the opening of the inner container 46, and the sponge 47 is used to The rat skin 45 is used for support, and the inner container 46 is received in the outer container 44.
  • the outer container 44 is used to collect the liquid flowing out of the inner container 46 to avoid contaminating the external environment.
  • curve a1 represents the response curve corresponding to mode a under 150V pulse voltage
  • curve b1 represents the response corresponding to mode b under 150V pulse voltage
  • curve c1 represents the response curve corresponding to mode c under 150V pulse voltage
  • curve d1 represents the response curve corresponding to mode d under 150V pulse voltage
  • curve e1 represents the response curve corresponding to mode e under 150V pulse voltage.
  • the electroporation process is performed using a pulse voltage of 300V, the number of pulses is 100, and the set pulse width and period.
  • the above five extraction methods are used to perform reverse electroosmosis extraction to obtain the liquid to be tested. , detect the liquid to be tested, and obtain the DPV response curve shown in Figure 6, where curve a2 represents the response curve corresponding to mode a under 300V pulse voltage, curve b2 represents the response curve corresponding to mode b under 300V pulse voltage, and curve c2 represents 300V The response curve corresponding to mode c under pulse voltage.
  • Curve d2 represents the response curve corresponding to mode d under pulse voltage of 300V.
  • Curve e2 represents the response curve corresponding to mode e under pulse voltage of 300V.
  • Figures 5 and 6 show the test results corresponding to the optimal pulse parameters and current values obtained experimentally, and the results of the remaining pulse parameters and current values are not shown. It can be seen from Figure 5 and Figure 6 that under the same pulse voltage, the extraction amount of biochemical substances increases with the increase of extraction time. Higher pulse voltage can obtain more extraction amount. The holes formed by electroporation have an impact on the extraction amount. Significant impact.
  • Figure 7 is a schematic diagram of the extraction efficiency at different extraction times when 150V pulse voltage is used for electroporation treatment, 300V pulse voltage is used for electroporation treatment, and no electroporation treatment is performed.
  • Table 1 it can be seen that when the extraction time reaches At 10 minutes, the penetration-promoting multiples of 150V pulse voltage and 300V pulse voltage can both reach 8 times, indicating that electroporation combined with reverse electroosmosis can effectively improve the transdermal extraction efficiency of macromolecule substances.
  • FIG 8 is a schematic flow chart of a biochemical substance detection method according to the second embodiment. As shown in Figure 8, this application also provides a biochemical substance detection method, which is applied to the biochemical substance detection system described in the above embodiment. The method includes:
  • Step S1 Apply a pulse voltage to the detection area through the first electrode group and the second electrode group to perform electroporation treatment on the detection area;
  • Step S2 Apply a constant current to the detection area through the first electrode group and the second electrode group, so that the biochemical substances are extracted into the electrolyte in the area where the first working electrode and the second working electrode are located;
  • Step S3 Detect the biochemical substance through the first working electrode and/or the second working electrode to determine the concentration of the biochemical substance.
  • the first electrode group further includes a first pair of electrodes, a first reference electrode, a first extraction electrode and a first flexible substrate, a first working electrode, a first pair of electrodes, a first reference electrode and a first flexible substrate.
  • An extraction electrode is disposed on the first flexible substrate;
  • the second electrode group also includes a second counter electrode, a second reference electrode, a second extraction electrode and a second flexible substrate.
  • the second working electrode, the second counter electrode, the second reference electrode and the second extraction electrode are arranged on the first Two flexible substrates;
  • the method includes applying pulse voltage and constant current to the detection area through the first extraction electrode and the second extraction electrode.
  • step S1 detects the biochemical substance through the first working electrode and/or the second working electrode, and determines the concentration of the biochemical substance, including:
  • Differential pulse voltammetry is used to determine the concentration of biochemical substances based on the electrochemical signals generated by the first working electrode and/or the second working electrode.
  • the pulse parameters for applying the pulse voltage include pulse voltage, pulse width, period and number of pulses.
  • the pulse voltage ranges from 100V to 500V
  • the pulse width ranges from 10 ms to 50 ms
  • the period ranges from 100V to 500V. is 0 s -20 s
  • the number of pulses ranges from 20 to 200.
  • the current parameters used to apply a constant current include a current value and an extraction time.
  • the current value ranges from 200uA to 500uA, and the extraction time ranges from 1 min to 10 min.
  • the first biosensing layer and the second biosensing layer are used to detect different biochemical substances, and the biochemical substances include luteinizing hormone and follicle-stimulating hormone.
  • This application also provides a computer-readable storage medium.
  • a computer program is stored on the computer-readable storage medium.
  • the computer program is executed by a processor, the steps of the biochemical substance detection method described in the above embodiments are implemented.

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Abstract

The present application relates to a biochemical substance measurement system and method, and a storage medium. The system comprises a first electrode group, a second electrode group and a treatment device, wherein the first electrode group comprises a first working electrode, a surface of which is modified with a first biological sensing layer; the second electrode group comprises a second working electrode, a surface of which is modified with a second biological sensing layer; and the treatment device is configured to: apply pulse voltages to a measurement area by means of the first electrode group and the second electrode group, so as to perform an electroporation treatment; apply constant currents to the measurement area by means of the first electrode group and the second electrode group, so as to extract a biochemical substance to electrolytes in areas where the first working electrode and the second working electrode are located; and measure the biochemical substance by means of the first working electrode and/or the second working electrode, so as to determine the concentration of the biochemical substance. By means of the technical solution in the present application, an electroporation treatment is first performed on a measurement area, and a biochemical substance is then extracted, such that the transdermal extraction efficiency of the biochemical substance can be improved; and the technical solution is more suitable for the extraction and measurement of a macromolecular substance.

Description

一种生化物质检测系统、方法及存储介质A biochemical substance detection system, method and storage medium 技术领域Technical field
本申请涉及生化物质检测技术领域,具体涉及一种生化物质检测系统、方法及存储介质。This application relates to the technical field of biochemical substance detection, and specifically relates to a biochemical substance detection system, method and storage medium.
背景技术Background technique
人体的正常生命活动离不开体内各种生化物质的刺激和调节作用,一种或几种生化指标的变化会影响到人体的正常运行甚至会影响生命健康。例如,人体的血糖浓度水平与波动是糖代谢异常患者的重要监测和疾病管理指标;促黄体生成素作为性激素检测的重要指标之一,其浓度水平可用于指导诊断和疗效评价等。The normal life activities of the human body are inseparable from the stimulation and regulation of various biochemical substances in the body. Changes in one or several biochemical indicators will affect the normal operation of the human body and even affect life and health. For example, the human body's blood glucose concentration levels and fluctuations are important monitoring and disease management indicators for patients with abnormal glucose metabolism; luteinizing hormone is one of the important indicators for sex hormone detection, and its concentration level can be used to guide diagnosis and efficacy evaluation.
通过透皮提取技术将体内的生化物质提取到体外进行检测,可以避免疼痛和对皮肤的伤害,有效规避了伤口感染的风险,同时对患者体内物质进行频繁采集和连续监测也具有积极的意义。目前,常用的无创提取方式有尿液提取、汗液提取和反电渗提取等,其中反电渗提取的方式更有利于随时监测和及时监测。反电渗提取是在离子导入和反向离子渗透双重作用下,将血液和组织液中的目标分子迁移流动到皮肤表面进行测量,但此方法的提取效率有限,且不利于大分子物质的提取与检测。Through transdermal extraction technology, biochemical substances in the body are extracted outside the body for detection, which can avoid pain and damage to the skin, and effectively avoid the risk of wound infection. At the same time, frequent collection and continuous monitoring of substances in the patient's body are also of positive significance. Currently, commonly used non-invasive extraction methods include urine extraction, sweat extraction and reverse electroosmosis extraction, among which the reverse electroosmosis extraction method is more conducive to anytime and timely monitoring. Reverse electroosmosis extraction uses the dual action of ion introduction and reverse ion osmosis to migrate target molecules in blood and tissue fluid to the skin surface for measurement. However, the extraction efficiency of this method is limited and is not conducive to the extraction and extraction of macromolecular substances. detection.
技术解决方案Technical solutions
针对上述技术问题,本申请提供一种生化物质检测系统、方法及存储介质,先对检测区域进行电穿孔处理再提取生化物质,能够提高生化物质的透皮提取效率,且更适用于大分子物质的提取与检测。In view of the above technical problems, this application provides a biochemical substance detection system, method and storage medium. The detection area is electroporated first and then the biochemical substances are extracted. This can improve the transdermal extraction efficiency of biochemical substances and is more suitable for macromolecular substances. extraction and detection.
本申请提供一种生化物质检测系统,包括第一电极组、第二电极组与处理设备,所述处理设备与所述第一电极组、所述第二电极组连接;The present application provides a biochemical substance detection system, including a first electrode group, a second electrode group and a processing device, and the processing device is connected to the first electrode group and the second electrode group;
所述第一电极组包括表面修饰有第一生物传感层的第一工作电极,所述第二电极组包括表面修饰有第二生物传感层的第二工作电极,所述第一生物传感层与所述第二生物传感层用于检测相同或不同的生化物质;The first electrode group includes a first working electrode with a first biosensing layer modified on its surface, and the second electrode group includes a second working electrode with a second biosensing layer modified on its surface. The sensing layer and the second biosensing layer are used to detect the same or different biochemical substances;
所述处理设备被配置成用于:The processing equipment is configured for:
通过所述第一电极组与所述第二电极组向检测区域施加脉冲电压,以对所述检测区域进行电穿孔处理;Apply a pulse voltage to the detection area through the first electrode group and the second electrode group to perform electroporation treatment on the detection area;
通过所述第一电极组与所述第二电极组向所述检测区域施加恒电流,以使生化物质被提取至所述第一工作电极与所述第二工作电极所在区域的电解液;Apply a constant current to the detection area through the first electrode group and the second electrode group, so that biochemical substances are extracted into the electrolyte in the area where the first working electrode and the second working electrode are located;
通过所述第一工作电极和/或所述第二工作电极对所述生化物质进行检测,确定所述生化物质的浓度。The biochemical substance is detected through the first working electrode and/or the second working electrode to determine the concentration of the biochemical substance.
在一个实施例中,所述第一电极组还包括第一对电极、第一参比电极、第一提取电极及第一柔性基底,所述第一工作电极、第一对电极、第一参比电极及第一提取电极设置在所述第一柔性基底上;In one embodiment, the first electrode group further includes a first pair of electrodes, a first reference electrode, a first extraction electrode and a first flexible substrate, and the first working electrode, the first pair of electrodes, the first reference electrode The specific electrode and the first extraction electrode are arranged on the first flexible substrate;
所述第二电极组还包括第二对电极、第二参比电极、第二提取电极及第二柔性基底,所述第二工作电极、第二对电极、第二参比电极及第二提取电极设置在所述第二柔性基底上;The second electrode group also includes a second counter electrode, a second reference electrode, a second extraction electrode and a second flexible substrate. The second working electrode, the second counter electrode, the second reference electrode and the second extraction electrode Electrodes are provided on the second flexible substrate;
其中,所述第一提取电极和所述第二提取电极用于向所述检测区域施加所述脉冲电压和所述恒电流。Wherein, the first extraction electrode and the second extraction electrode are used to apply the pulse voltage and the constant current to the detection area.
在一个实施例中,所述第一工作电极为片状电极,所述第一对电极与所述第一参比电极环绕所述第一工作电极的外周设置,所述第一提取电极环绕所述第一对电极的外周设置;In one embodiment, the first working electrode is a sheet electrode, the first pair of electrodes and the first reference electrode are arranged around the periphery of the first working electrode, and the first extraction electrode surrounds The peripheral arrangement of the first pair of electrodes;
所述第二工作电极为片状电极,所述第二对电极与所述第二参比电极环绕所述第二工作电极的外周,所述第二提取电极环绕所述第二对电极的外周。The second working electrode is a sheet electrode, the second pair of electrodes and the second reference electrode surround the outer periphery of the second working electrode, and the second extraction electrode surrounds the outer periphery of the second pair of electrodes. .
在一个实施例中,所述第一电极组设有第一支撑环,所述第一支撑环环绕所述第一电极组中的电极的外周设置;In one embodiment, the first electrode group is provided with a first support ring, and the first support ring is arranged around the periphery of the electrodes in the first electrode group;
所述第二电极组设有第二支撑环,所述第二支撑环环绕所述第二电极组中的电极的外周设置。The second electrode group is provided with a second support ring, and the second support ring is arranged around the periphery of the electrodes in the second electrode group.
在一个实施例中,通过所述第一工作电极和/或所述第二工作电极对所述生化物质进行检测,确定所述生化物质的浓度时,所述处理设备被配置成用于:In one embodiment, when the biochemical substance is detected through the first working electrode and/or the second working electrode, and the concentration of the biochemical substance is determined, the processing device is configured to:
采用差分脉冲伏安法,根据所述第一工作电极和/或所述第二工作电极产生的电化学信号,确定所述生化物质的浓度。Differential pulse voltammetry is used to determine the concentration of the biochemical substance based on the electrochemical signals generated by the first working electrode and/or the second working electrode.
在一个实施例中,用于施加所述脉冲电压的脉冲参数包括脉冲电压、脉宽、周期与脉冲个数,所述脉冲电压的范围为100V -500V,所述脉宽的范围为10 ms -50 ms,所述周期的范围为0 s -20 s,所述脉冲个数的范围为20个-200个。In one embodiment, the pulse parameters for applying the pulse voltage include pulse voltage, pulse width, period and number of pulses, the range of the pulse voltage is 100V -500V, and the range of the pulse width is 10 ms - 50 ms, the period ranges from 0 s to 20 s, and the number of pulses ranges from 20 to 200.
在一个实施例中,用于施加所述恒电流的电流参数包括电流值与提取时间,所述电流值的范围为200μA -500μA,所述提取时间的范围为1 min -10 min。In one embodiment, the current parameters used to apply the constant current include a current value and an extraction time. The current value ranges from 200 μA to 500 μA, and the extraction time ranges from 1 min to 10 min.
在一个实施例中,所述第一生物传感层与所述第二生物传感层用于检测不同的生化物质,所述生化物质包括促黄体生成激素与促卵泡素。In one embodiment, the first biosensing layer and the second biosensing layer are used to detect different biochemical substances, and the biochemical substances include luteinizing hormone and follicle-stimulating hormone.
本申请还提供一种生化物质检测方法,应用于如上所述的生化物质检测系统,所述方法包括:This application also provides a biochemical substance detection method, which is applied to the biochemical substance detection system as described above. The method includes:
通过所述第一电极组与所述第二电极组向检测区域施加脉冲电压,以对所述检测区域进行电穿孔处理;Apply a pulse voltage to the detection area through the first electrode group and the second electrode group to perform electroporation treatment on the detection area;
通过所述第一电极组与所述第二电极组向所述检测区域施加恒电流,以使生化物质被提取至所述第一工作电极与所述第二工作电极所在区域的电解液;Apply a constant current to the detection area through the first electrode group and the second electrode group, so that biochemical substances are extracted into the electrolyte in the area where the first working electrode and the second working electrode are located;
通过所述第一工作电极和/或所述第二工作电极对所述生化物质进行检测,确定所述生化物质的浓度。The biochemical substance is detected through the first working electrode and/or the second working electrode to determine the concentration of the biochemical substance.
本申请还提供一种计算机可读存储介质,所述计算机可读存储介质上存储计算机程序,所述计算机程序被处理器执行时实现如上所述的生化物质检测方法的步骤。The present application also provides a computer-readable storage medium. A computer program is stored on the computer-readable storage medium. When the computer program is executed by a processor, the steps of the biochemical substance detection method as described above are implemented.
本申请的生化物质检测系统、方法及存储介质,系统包括第一电极组、第二电极组与处理设备;第一电极组包括表面修饰有第一生物传感层的第一工作电极,第二电极组包括表面修饰有第二生物传感层的第二工作电极;处理设备被配置成用于:通过第一电极组与第二电极组向检测区域施加脉冲电压,进行电穿孔处理;通过第一电极组与第二电极组向检测区域施加恒电流,将生化物质提取至第一工作电极与第二工作电极所在区域的电解液;通过第一工作电极和/或第二工作电极对生化物质进行检测,确定生化物质的浓度。本申请的技术方案,先对检测区域进行电穿孔处理再提取生化物质,能够提高生化物质的透皮提取效率,且更适用于大分子物质的提取与检测。In the biochemical substance detection system, method and storage medium of the present application, the system includes a first electrode group, a second electrode group and processing equipment; the first electrode group includes a first working electrode with a first biosensing layer modified on its surface, and a second The electrode group includes a second working electrode with a second biosensing layer modified on its surface; the processing device is configured to: apply a pulse voltage to the detection area through the first electrode group and the second electrode group to perform electroporation processing; An electrode group and a second electrode group apply a constant current to the detection area to extract biochemical substances into the electrolyte in the area where the first working electrode and the second working electrode are located; the biochemical substances are extracted through the first working electrode and/or the second working electrode. Tests are performed to determine the concentration of biochemical substances. The technical solution of this application first electroporates the detection area and then extracts biochemical substances, which can improve the transdermal extraction efficiency of biochemical substances and is more suitable for the extraction and detection of macromolecular substances.
附图说明Description of the drawings
图1是根据第一实施例示出的生化物质检测系统的结构示意图。Figure 1 is a schematic structural diagram of a biochemical substance detection system according to the first embodiment.
图2是电穿孔通道的示意图。Figure 2 is a schematic diagram of the electroporation channel.
图3是根据第一实施例示出的生化物质检测的一种实验设备的示意图。Figure 3 is a schematic diagram of an experimental device for biochemical substance detection according to the first embodiment.
图4是图3所示的实验设备的剖视图。FIG. 4 is a cross-sectional view of the experimental equipment shown in FIG. 3 .
图5是采用150V脉冲电压和不同提取时间对应的提取液的DPV响应曲线。Figure 5 is the DPV response curve of the extraction solution using 150V pulse voltage and different extraction times.
图6是采用300V脉冲电压和不同提取时间对应的提取液的DPV响应曲线。Figure 6 is the DPV response curve of the extraction solution using 300V pulse voltage and different extraction times.
图7是不同处理方式的提取结果示意图。Figure 7 is a schematic diagram of the extraction results of different processing methods.
图8是根据第二实施例示出的生化物质检测方法的流程示意图。Figure 8 is a schematic flow chart of a biochemical substance detection method according to the second embodiment.
本申请的实施方式Implementation Mode of this Application
以下由特定的具体实施例说明本申请的实施方式,熟悉此技术的人士可由本说明书所揭露的内容轻易地了解本申请的其他优点及功效。The following describes the implementation of the present application through specific embodiments. Those familiar with this technology can easily understand other advantages and effects of the present application from the content disclosed in this specification.
在下述描述中,参考附图,附图描述了本申请的若干实施例。应当理解,还可使用其他实施例,并且可以在不背离本申请的精神和范围的情况下进行机械组成、结构、电气以及操作上的改变。下面的详细描述不应该被认为是限制性的,这里使用的术语仅是为了描述特定实施例,而并非旨在限制本申请。In the following description, reference is made to the accompanying drawings, which illustrate several embodiments of the application. It is to be understood that other embodiments may be utilized and mechanical, structural, electrical, as well as operational changes may be made without departing from the spirit and scope of the present application. The following detailed description should not be considered limiting and the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the application.
虽然在一些实例中术语第一、第二等在本文中用来描述各种元件,但是这些元件不应当被这些术语限制。这些术语仅用来将一个元件与另一个元件进行区分。Although in some instances the terms first, second, etc. are used herein to describe various elements, these elements should not be limited by these terms. These terms are only used to distinguish one element from another element.
再者,如同在本文中所使用的,单数形式“一”、“一个”和“该”旨在也包括复数形式,除非上下文中有相反的指示。应当进一步理解,术语“包含”、“包括”表明存在所述的特征、步骤、操作、元件、组件、项目、种类、和/或组,但不排除一个或多个其他特征、步骤、操作、元件、组件、项目、种类、和/或组的存在、出现或添加。此处使用的术语“或”和“和/或”被解释为包括性的,或意味着任一个或任何组合。因此,“A、B或C”或者“A、B和/或C”意味着“以下任一个:A;B;C;A和B;A和C;B和C;A、B和C”。仅当元件、功能、步骤或操作的组合在某些方式下内在地互相排斥时,才会出现该定义的例外。Furthermore, as used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context indicates otherwise. It should be further understood that the terms "comprising" and "including" indicate the presence of stated features, steps, operations, elements, components, items, categories, and/or groups, but do not exclude one or more other features, steps, operations, The presence, occurrence, or addition of elements, components, items, categories, and/or groups. The terms "or" and "and/or" as used herein are to be construed as inclusive or to mean any one or any combination. Therefore, "A, B or C" or "A, B and/or C" means "any of the following: A; B; C; A and B; A and C; B and C; A, B and C" . Exceptions to this definition occur only when the combination of elements, functions, steps, or operations is inherently mutually exclusive in some manner.
第一实施例First embodiment
图1是根据第一实施例示出的生化物质检测系统的结构示意图。如图1所示,本实施例的生化物质检测系统,包括第一电极组10、第二电极组20与处理设备30,处理设备30与第一电极组10、第二电极组20连接。Figure 1 is a schematic structural diagram of a biochemical substance detection system according to the first embodiment. As shown in FIG. 1 , the biochemical substance detection system of this embodiment includes a first electrode group 10 , a second electrode group 20 and a processing device 30 . The processing device 30 is connected to the first electrode group 10 and the second electrode group 20 .
第一电极组10包括表面修饰有第一生物传感层的第一工作电极11,第二电极组20包括表面修饰有第二生物传感层的第二工作电极21,第一生物传感层与第二生物传感层用于检测相同或不同的生化物质。The first electrode group 10 includes a first working electrode 11 with a first biosensing layer modified on its surface. The second electrode group 20 includes a second working electrode 21 with a second biosensing layer modified on its surface. The first biosensing layer and the second biosensing layer for detecting the same or different biochemical substances.
处理设备30被配置成用于:通过第一电极组10与第二电极组20向检测区域施加脉冲电压,以对检测区域进行电穿孔处理;通过第一电极组10与第二电极组20向检测区域施加恒电流,以使生化物质被提取至第一工作电极11与第二工作电极21所在区域的电解液;通过第一工作电极11和/或第二工作电极21对生化物质进行检测,确定生化物质的浓度。The processing device 30 is configured to: apply a pulse voltage to the detection area through the first electrode group 10 and the second electrode group 20 to perform electroporation treatment on the detection area; A constant current is applied to the detection area so that the biochemical substances are extracted into the electrolyte in the area where the first working electrode 11 and the second working electrode 21 are located; the biochemical substances are detected through the first working electrode 11 and/or the second working electrode 21, Determine the concentration of biochemical substances.
其中,处理设备30为大型或小型电化学工作站,同时能够提供电穿孔所需的脉冲电压,第一电极组10、第二电极组20可采用插接的方式与处理设备30连接。The processing equipment 30 is a large or small electrochemical workstation and can provide the pulse voltage required for electroporation. The first electrode group 10 and the second electrode group 20 can be connected to the processing equipment 30 in a plug-in manner.
第一工作电极11与第二工作电极21可以用于向检测区域施加脉冲电压、施加恒电流及对生化物质进行检测,以简化电极组的结构。或者,向检测区域施加脉冲电压、施加恒电流可以通过第一电极组10、第二电极组20中的其他电极进行,以提高第一工作电极11与第二工作电极21的使用寿命。The first working electrode 11 and the second working electrode 21 can be used to apply pulse voltage and constant current to the detection area and detect biochemical substances to simplify the structure of the electrode group. Alternatively, applying pulse voltage and constant current to the detection area can be performed through other electrodes in the first electrode group 10 and the second electrode group 20 to increase the service life of the first working electrode 11 and the second working electrode 21 .
可选地,通过在第一工作电极11与第二工作电极21修饰相同或不同的生物传感层,实现对相同或不同的生化物质的检测。可选地,第一生物传感层和第二生物传感层用于检测不同的大分子生化物质,大分子生化物质包括促黄体生成激素与促卵泡素。以促黄体生成激素为例,通过在第一工作电极11的表面修饰氧化石墨烯/硫堇/纳米金复合物、促黄体生成激素抗体、牛血清白蛋白作为第一生物传感层,可以实现对促黄体生成激素进行检测。Optionally, the same or different biochemical substances can be detected by modifying the same or different biosensing layers on the first working electrode 11 and the second working electrode 21 . Optionally, the first biosensing layer and the second biosensing layer are used to detect different macromolecular biochemical substances, and the macromolecular biochemical substances include luteinizing hormone and follicle-stimulating hormone. Taking luteinizing hormone as an example, by modifying the surface of the first working electrode 11 with a graphene oxide/thionine/gold nanocomposite, luteinizing hormone antibody, and bovine serum albumin as the first biosensing layer, it can be achieved Testing for luteinizing hormone.
通过第一电极组10与第二电极组20向检测区域施加恒电流,即进行反电渗提取。反电渗提取过程中,在皮肤表面施加电场使皮下组织中的Cl -和Na +在电场作用下分别向正负极移动,形成一个从皮肤表层经过皮下组织再回到皮肤表层的直流电流通道,利用这个离子流可以将皮下组织液中的生化物质携带到皮肤表面进行检测,但由于皮肤屏障的存在,此方式的提取效率有限,且大分子物质由于直径过大也较难通过反电渗的方式提取到皮肤表面。对此,本申请在进行反电渗提取之前,先采用合适的脉冲参数对检测区域进行电穿孔处理,以此提高生化物质的透皮提取效率,且更适用于大分子物质的提取与检测。 A constant current is applied to the detection area through the first electrode group 10 and the second electrode group 20, that is, reverse electroosmosis extraction is performed. During the reverse electroosmosis extraction process, an electric field is applied to the skin surface to cause Cl - and Na + in the subcutaneous tissue to move toward the positive and negative electrodes respectively under the action of the electric field, forming a DC current channel from the skin surface through the subcutaneous tissue and back to the skin surface. , using this ion current, biochemical substances in subcutaneous tissue fluid can be carried to the skin surface for detection. However, due to the existence of the skin barrier, the extraction efficiency of this method is limited, and it is difficult for macromolecular substances to pass through reverse electroosmosis due to their large diameter. extracted to the skin surface. In this regard, before performing reverse electroosmosis extraction, this application first uses appropriate pulse parameters to electroporate the detection area, thereby improving the transdermal extraction efficiency of biochemical substances and making it more suitable for the extraction and detection of macromolecular substances.
请参考图2,电穿孔处理过程中,通过穿过非导电的角质层建立一个穿过皮肤的占优势的电压梯度,若电压梯度超过了屏障击穿电位,就会形成孔洞,使得物质可以穿过皮肤屏障进行传输,例如通过毛囊A、角质细胞间隙B、汗腺C、其他孔道D等通道从皮下转移到皮肤表面。根据使用的脉冲参数,这些通道会在一段时间后重新封闭或恢复原始状态。在皮肤表面形成可逆通道后再进行反电渗提取,能够从组织液和血液中提取出更多的目标物质,有效提高皮下目标物质的提取量,同时,也有利于提高提取效率,并提取出大分子物质,实现大分子物质的检测。Please refer to Figure 2. During the electroporation process, a dominant voltage gradient across the skin is established by passing through the non-conductive stratum corneum. If the voltage gradient exceeds the barrier breakdown potential, holes will be formed, allowing substances to penetrate Transmitted through the skin barrier, for example, from the subcutaneous to the skin surface through channels such as hair follicles A, keratinocyte gaps B, sweat glands C, and other pores D. Depending on the pulse parameters used, these channels will re-close after a period of time or return to their original state. After forming a reversible channel on the skin surface and then performing reverse electroosmosis extraction, more target substances can be extracted from tissue fluid and blood, effectively increasing the extraction amount of subcutaneous target substances. At the same time, it is also conducive to improving extraction efficiency and extracting large amounts of target substances. Molecular substances, realizing the detection of macromolecular substances.
在一个实施例中,第一电极组10还包括第一对电极12、第一参比电极13、第一提取电极14及第一柔性基底(图1中未示出),第一工作电极11、第一对电极12、第一参比电极13及第一提取电极14设置在第一柔性基底上。第二电极组20还包括第二对电极22、第二参比电极23、第二提取电极14及第二柔性基底(图1中未示出),第二工作电极21、第二对电极22、第二参比电极23及第二提取电极14设置在第二柔性基底上。第一柔性基底与第二柔性基底上设有导线与接口,各电极通过导线分别与接口连接,接口用于连接处理设备30,导线在柔性基底上的排布方式包括折线排布、直线排布、曲线排布等多种排布方式。可选地,第一工作电极11与第二工作电极21的材料为Pt/C,其余电极的材料可均为Ag/AgCl。柔性基底可完全贴合体表,能够满足可穿戴、易携带的检测需求。In one embodiment, the first electrode group 10 further includes a first pair of electrodes 12 , a first reference electrode 13 , a first extraction electrode 14 and a first flexible substrate (not shown in FIG. 1 ). The first working electrode 11 , the first pair of electrodes 12, the first reference electrode 13 and the first extraction electrode 14 are arranged on the first flexible substrate. The second electrode group 20 also includes a second counter electrode 22 , a second reference electrode 23 , a second extraction electrode 14 and a second flexible substrate (not shown in FIG. 1 ). The second working electrode 21 and the second counter electrode 22 , the second reference electrode 23 and the second extraction electrode 14 are arranged on the second flexible substrate. Wires and interfaces are provided on the first flexible substrate and the second flexible substrate. Each electrode is connected to the interface through a wire. The interface is used to connect the processing device 30. The arrangement of the wires on the flexible substrate includes a zigzag arrangement and a straight line arrangement. , curve arrangement and other arrangement methods. Optionally, the materials of the first working electrode 11 and the second working electrode 21 are Pt/C, and the materials of the remaining electrodes can be Ag/AgCl. The flexible base can completely fit the body surface and meet the needs of wearable and easy-to-carry testing.
在一个实施例中,第一工作电极11为片状电极,第一对电极12与第一参比电极13环绕第一工作电极11的外周设置,第一提取电极14环绕第一对电极12的外周设置。具体地,第一工作电极11的位于检测区域的部分为圆片状,第一对电极12与第一参比电极13共同环绕于第一工作电极11的圆片状部分的外周,即,环绕第一工作电极11的圆片状部的外周的一圈区域中,部分区域设置第一对电极12,另部分区域设置第一参比电极13,以此减小电极的尺寸。第一提取电极14环绕第一对电极12的外周,即设置在第一电极组10中其他电极的最外侧。In one embodiment, the first working electrode 11 is a sheet electrode, the first pair of electrodes 12 and the first reference electrode 13 are arranged around the periphery of the first working electrode 11 , and the first extraction electrode 14 surrounds the periphery of the first pair of electrodes 12 Peripheral settings. Specifically, the part of the first working electrode 11 located in the detection area is in the shape of a disk, and the first counter electrode 12 and the first reference electrode 13 are commonly surrounding the outer periphery of the disk-shaped part of the first working electrode 11 , that is, surrounding In a circumferential area around the disk-shaped portion of the first working electrode 11, a first pair of electrodes 12 is provided in some areas, and a first reference electrode 13 is provided in another area, thereby reducing the size of the electrodes. The first extraction electrode 14 surrounds the outer periphery of the first pair of electrodes 12 , that is, is disposed on the outermost side of other electrodes in the first electrode group 10 .
第二工作电极21为片状电极,第二对电极22与第二参比电极23环绕第二工作电极21的外周,第二提取电极14环绕第二对电极22的外周。第一电极组10与第二电极组20中的电极的设置方式相同,不再进行赘述。The second working electrode 21 is a sheet electrode, the second counter electrode 22 and the second reference electrode 23 surround the outer periphery of the second working electrode 21 , and the second extraction electrode 14 surrounds the outer periphery of the second counter electrode 22 . The electrodes in the first electrode group 10 and the second electrode group 20 are arranged in the same manner, which will not be described again.
对电极用于与相应的工作电极构成极化回路,以使工作电极有电流通过,参比电极用于在检测生化物质的过程中,提供并保持一个固定的参比电势。第一提取电极14和第二提取电极14用于向检测区域施加脉冲电压以进行电穿孔处理,还用于向检测区域施加恒电流以进行反电渗提取。The counter electrode is used to form a polarization loop with the corresponding working electrode so that current can flow through the working electrode. The reference electrode is used to provide and maintain a fixed reference potential during the detection of biochemical substances. The first extraction electrode 14 and the second extraction electrode 14 are used to apply pulse voltage to the detection area to perform electroporation treatment, and are also used to apply constant current to the detection area to perform reverse electroosmosis extraction.
在一个实施例中,用于施加脉冲电压的脉冲参数包括脉冲电压、脉宽、周期与脉冲个数,可选地,脉冲电压的范围为100V -500V,脉宽的范围为10 ms -50 ms,周期的范围为0 s -20 s,脉冲个数的范围为20个-200个。在一个实施例中,用于施加恒电流的电流参数包括电流值与提取时间,电流值的范围为200μA-500μA,可选地,提取时间的范围为1 min -10 min。优选地,脉冲电压为150 V或300 V、脉冲个数为100个、恒电流300 μA、提取时间为10 min时,提取效率最高。In one embodiment, the pulse parameters for applying the pulse voltage include pulse voltage, pulse width, period and number of pulses. Optionally, the pulse voltage ranges from 100V to 500V, and the pulse width ranges from 10 ms to 50 ms. , the period ranges from 0 s to 20 s, and the number of pulses ranges from 20 to 200. In one embodiment, the current parameters for applying a constant current include a current value and an extraction time. The current value ranges from 200 μA to 500 μA. Alternatively, the extraction time ranges from 1 min to 10 min. Preferably, the extraction efficiency is highest when the pulse voltage is 150 V or 300 V, the number of pulses is 100, the constant current is 300 μA, and the extraction time is 10 minutes.
在一个实施例中,第一电极组10设有第一支撑环(图1中未示出),第一支撑环环绕第一电极组10中的电极的外周设置,即位于第一提取电极14的外周。第二电极组20设有第二支撑环(图1中未示出),第二支撑环环绕第二电极组20中的电极的外周设置,即位于第二提取电极14的外周。第一支撑环与第二支撑环可以是具有一定厚度的环形胶贴,胶贴的一侧与柔性基底贴附,另一侧用于与体表贴附,当第一电极组10与第二电极组20贴附在体表时,通过支撑环可以在电极与体表之间形成用于存储电解液的储液空间,在对检测区域施加脉冲电压、施加恒电流及检测生化物质浓度时,形成电极与体表之间的电通路,同时,提取的生化物质在电解液中扩散以处于工作电极的所在区域,实现检测。In one embodiment, the first electrode group 10 is provided with a first support ring (not shown in FIG. 1 ), which is disposed around the periphery of the electrodes in the first electrode group 10 , that is, located at the first extraction electrode 14 of the periphery. The second electrode group 20 is provided with a second support ring (not shown in FIG. 1 ). The second support ring is arranged around the periphery of the electrodes in the second electrode group 20 , that is, located on the periphery of the second extraction electrode 14 . The first support ring and the second support ring may be annular adhesive tapes with a certain thickness. One side of the adhesive tape is attached to the flexible base, and the other side is used to adhere to the body surface. When the first electrode group 10 and the second When the electrode set 20 is attached to the body surface, a storage space for storing electrolyte can be formed between the electrodes and the body surface through the support ring. When applying pulse voltage and constant current to the detection area and detecting the concentration of biochemical substances, An electrical path is formed between the electrode and the body surface. At the same time, the extracted biochemical substances diffuse in the electrolyte to be in the area of the working electrode to achieve detection.
在一个实施例中,通过第一工作电极11和/或第二工作电极21对生化物质进行检测,确定生化物质的浓度时,处理设备30被配置成用于:In one embodiment, when detecting biochemical substances through the first working electrode 11 and/or the second working electrode 21 and determining the concentration of the biochemical substances, the processing device 30 is configured to:
采用差分脉冲伏安法,根据第一工作电极11和/或第二工作电极21产生的电化学信号,确定生化物质的浓度。Differential pulse voltammetry is used to determine the concentration of biochemical substances based on the electrochemical signals generated by the first working electrode 11 and/or the second working electrode 21 .
其中,采用差分脉冲伏安法,通过处理设备30采集第一工作电极11、第二工作电极21的电化学信号,第一工作电极11、第二工作电极21的电化学信号的变化与待测液中生化物质的浓度成比例关系,从而,处理设备30可以根据第一工作电极11、第二工作电极21的电化学信号的变化,确定待测液中的生化物质的浓度。Among them, differential pulse voltammetry is used to collect electrochemical signals of the first working electrode 11 and the second working electrode 21 through the processing device 30. The changes of the electrochemical signals of the first working electrode 11 and the second working electrode 21 are related to those to be measured. The concentration of biochemical substances in the liquid is proportional. Therefore, the processing device 30 can determine the concentration of the biochemical substances in the liquid to be tested based on changes in the electrochemical signals of the first working electrode 11 and the second working electrode 21 .
通过上述方式,本申请先对检测区域进行电穿孔处理再提取生化物质,能够提高生化物质的透皮提取效率,且更适用于大分子物质的提取与检测。以下通过实验数据对本申请的方案进行说明。Through the above method, this application first electroporates the detection area and then extracts biochemical substances, which can improve the transdermal extraction efficiency of biochemical substances and is more suitable for the extraction and detection of macromolecular substances. The scheme of this application is explained below through experimental data.
请结合图3与图4,一种实验设备包括电极组40、电路板41、电阻触点42、基座43、鼠皮45、海绵47、内容器46、外容器44。其中,电极组40包括第一电极组10与第二电极组20,电路板41与电极组连接并设置有电阻触点42,电阻触点42用于与处理设备30连接,基座43用于支撑电路板41,内容器46用于盛装用于替代组织液的溶液,内容器46中放置海绵47,再将用于模拟体表的鼠皮45覆盖在内容器46的开口处,通过海绵47对鼠皮45进行支撑,内容器46收容于外容器44中,外容器44用于收集从内容器46中流出的液体以避免污染外部环境。Please combine Figure 3 and Figure 4. An experimental equipment includes an electrode group 40, a circuit board 41, a resistance contact 42, a base 43, rat skin 45, a sponge 47, an inner container 46, and an outer container 44. The electrode group 40 includes a first electrode group 10 and a second electrode group 20 . The circuit board 41 is connected to the electrode group and is provided with resistance contacts 42 . The resistance contacts 42 are used to connect to the processing equipment 30 , and the base 43 is used to connect to the processing equipment 30 . The circuit board 41 is supported, and the inner container 46 is used to hold a solution for replacing tissue fluid. A sponge 47 is placed in the inner container 46, and then the rat skin 45 used to simulate the body surface is covered at the opening of the inner container 46, and the sponge 47 is used to The rat skin 45 is used for support, and the inner container 46 is received in the outer container 44. The outer container 44 is used to collect the liquid flowing out of the inner container 46 to avoid contaminating the external environment.
搭建上述实验设备后,进行检测促黄体生成激素的实验。采用150V的脉冲电压、脉冲个数为100个及设定的脉宽、周期进行电穿孔处理,结束电穿孔处理后,采用以下五种提取方式进行反电渗提取,获取待测液:After setting up the above experimental equipment, conduct an experiment to detect luteinizing hormone. Use a pulse voltage of 150V, the number of pulses is 100, and the set pulse width and cycle are used for electroporation processing. After the electroporation processing is completed, the following five extraction methods are used for reverse electroosmosis extraction to obtain the liquid to be tested:
a. 在电穿孔的孔洞封闭后,以300 μA恒电流进行反电渗提取;a. After the electroporation holes are sealed, perform reverse electroosmosis extraction with a constant current of 300 μA;
b. 在电穿孔后,即以300 μA恒电流提取5min;b. After electroporation, extract at a constant current of 300 μA for 5 minutes;
c. 在电穿孔后,即以300 μA恒电流提取10min;c. After electroporation, extract at a constant current of 300 μA for 10 minutes;
d. 在电穿孔后,即以300 μA恒电流提取15min;d. After electroporation, extract at a constant current of 300 μA for 15 minutes;
e. 在电穿孔后,即以300 μA恒电流提取20min。e. After electroporation, extract at a constant current of 300 μA for 20 minutes.
检测上述五种提取方式获得的待测液,得到图5所示的DPV响应曲线,其中,曲线a1表示150V脉冲电压下a方式对应的响应曲线,曲线b1表示150V脉冲电压下b方式对应的响应曲线,曲线c1表示150V脉冲电压下c方式对应的响应曲线,曲线d1表示150V脉冲电压下d方式对应的响应曲线,曲线e1表示150V脉冲电压下e方式对应的响应曲线。Detect the liquid to be tested obtained by the above five extraction methods, and obtain the DPV response curve shown in Figure 5. Among them, curve a1 represents the response curve corresponding to mode a under 150V pulse voltage, and curve b1 represents the response corresponding to mode b under 150V pulse voltage. Curve, curve c1 represents the response curve corresponding to mode c under 150V pulse voltage, curve d1 represents the response curve corresponding to mode d under 150V pulse voltage, and curve e1 represents the response curve corresponding to mode e under 150V pulse voltage.
同样的,采用300V的脉冲电压、脉冲个数为100个及设定的脉宽、周期进行电穿孔处理,结束电穿孔处理后,采用上述五种提取方式进行反电渗提取,获取待测液,检测待测液,得到图6所示的DPV响应曲线,其中,曲线a2表示300V脉冲电压下a方式对应的响应曲线,曲线b2表示300V脉冲电压下b方式对应的响应曲线,曲线c2表示300V脉冲电压下c方式对应的响应曲线,曲线d2表示300V脉冲电压下d方式对应的响应曲线,曲线e2表示300V脉冲电压下e方式对应的响应曲线。Similarly, the electroporation process is performed using a pulse voltage of 300V, the number of pulses is 100, and the set pulse width and period. After the electroporation process is completed, the above five extraction methods are used to perform reverse electroosmosis extraction to obtain the liquid to be tested. , detect the liquid to be tested, and obtain the DPV response curve shown in Figure 6, where curve a2 represents the response curve corresponding to mode a under 300V pulse voltage, curve b2 represents the response curve corresponding to mode b under 300V pulse voltage, and curve c2 represents 300V The response curve corresponding to mode c under pulse voltage. Curve d2 represents the response curve corresponding to mode d under pulse voltage of 300V. Curve e2 represents the response curve corresponding to mode e under pulse voltage of 300V.
图5与图6所示为实验得到的最佳脉冲参数与电流值对应的测试结果,其余脉冲参数与电流值的结果未示出。从图5与图6可以看出,相同脉冲电压下,生化物质的提取量随提取时间的增加而增加,更高的脉冲电压可以获得更多的提取量,电穿孔形成的孔洞对提取量有显著影响。Figures 5 and 6 show the test results corresponding to the optimal pulse parameters and current values obtained experimentally, and the results of the remaining pulse parameters and current values are not shown. It can be seen from Figure 5 and Figure 6 that under the same pulse voltage, the extraction amount of biochemical substances increases with the increase of extraction time. Higher pulse voltage can obtain more extraction amount. The holes formed by electroporation have an impact on the extraction amount. Significant impact.
请参考图7,为分别采用150V脉冲电压进行电穿孔处理、300V脉冲电压进行电穿孔处理、不进行电穿孔处理时,不同提取时间的提取效率示意图,结合表1可以看出,当提取时间达到10 min时,150V脉冲电压与300V脉冲电压的促渗倍数均可达8倍,说明电穿孔结合反电渗的方式能够有效提升大分子物质的透皮提取效率。Please refer to Figure 7, which is a schematic diagram of the extraction efficiency at different extraction times when 150V pulse voltage is used for electroporation treatment, 300V pulse voltage is used for electroporation treatment, and no electroporation treatment is performed. Combined with Table 1, it can be seen that when the extraction time reaches At 10 minutes, the penetration-promoting multiples of 150V pulse voltage and 300V pulse voltage can both reach 8 times, indicating that electroporation combined with reverse electroosmosis can effectively improve the transdermal extraction efficiency of macromolecule substances.
表1. 促渗倍数对比表Table 1. Comparison table of penetration promotion multiples
提取时间/minExtraction time/min 150V脉冲电压150V pulse voltage 300V脉冲电压300V pulse voltage
55 5.35.3 6.46.4
1010 8.08.0 8.18.1
1515 2.52.5 2.72.7
2020 2.32.3 2.62.6
图8是根据第二实施例示出的生化物质检测方法的流程示意图。如图8所示,本申请还提供一种生化物质检测方法,应用于如上实施例所述的生化物质检测系统,方法包括:Figure 8 is a schematic flow chart of a biochemical substance detection method according to the second embodiment. As shown in Figure 8, this application also provides a biochemical substance detection method, which is applied to the biochemical substance detection system described in the above embodiment. The method includes:
步骤S1,通过第一电极组与第二电极组向检测区域施加脉冲电压,以对检测区域进行电穿孔处理;Step S1: Apply a pulse voltage to the detection area through the first electrode group and the second electrode group to perform electroporation treatment on the detection area;
步骤S2,通过第一电极组与第二电极组向检测区域施加恒电流,以使生化物质被提取至第一工作电极与第二工作电极所在区域的电解液;Step S2: Apply a constant current to the detection area through the first electrode group and the second electrode group, so that the biochemical substances are extracted into the electrolyte in the area where the first working electrode and the second working electrode are located;
步骤S3,通过第一工作电极和/或第二工作电极对生化物质进行检测,确定生化物质的浓度。Step S3: Detect the biochemical substance through the first working electrode and/or the second working electrode to determine the concentration of the biochemical substance.
在一个实施例中,第一电极组还包括第一对电极、第一参比电极、第一提取电极及第一柔性基底,第一工作电极、第一对电极、第一参比电极及第一提取电极设置在第一柔性基底上;In one embodiment, the first electrode group further includes a first pair of electrodes, a first reference electrode, a first extraction electrode and a first flexible substrate, a first working electrode, a first pair of electrodes, a first reference electrode and a first flexible substrate. An extraction electrode is disposed on the first flexible substrate;
第二电极组还包括第二对电极、第二参比电极、第二提取电极及第二柔性基底,第二工作电极、第二对电极、第二参比电极及第二提取电极设置在第二柔性基底上;The second electrode group also includes a second counter electrode, a second reference electrode, a second extraction electrode and a second flexible substrate. The second working electrode, the second counter electrode, the second reference electrode and the second extraction electrode are arranged on the first Two flexible substrates;
所述方法包括:通过第一提取电极和第二提取电极向检测区域施加脉冲电压和恒电流。The method includes applying pulse voltage and constant current to the detection area through the first extraction electrode and the second extraction electrode.
在一个实施例中,步骤S1通过第一工作电极和/或第二工作电极对生化物质进行检测,确定生化物质的浓度,包括:In one embodiment, step S1 detects the biochemical substance through the first working electrode and/or the second working electrode, and determines the concentration of the biochemical substance, including:
采用差分脉冲伏安法,根据第一工作电极和/或第二工作电极产生的电化学信号,确定生化物质的浓度。Differential pulse voltammetry is used to determine the concentration of biochemical substances based on the electrochemical signals generated by the first working electrode and/or the second working electrode.
在一个实施例中,用于施加脉冲电压的脉冲参数包括脉冲电压、脉宽、周期与脉冲个数,脉冲电压的范围为100V -500V,脉宽的范围为10 ms -50 ms,周期的范围为0 s -20 s,脉冲个数的范围为20个-200个。In one embodiment, the pulse parameters for applying the pulse voltage include pulse voltage, pulse width, period and number of pulses. The pulse voltage ranges from 100V to 500V, the pulse width ranges from 10 ms to 50 ms, and the period ranges from 100V to 500V. is 0 s -20 s, and the number of pulses ranges from 20 to 200.
在一个实施例中,用于施加恒电流的电流参数包括电流值与提取时间,电流值的范围为200uA -500uA,提取时间的范围为1 min -10 min。In one embodiment, the current parameters used to apply a constant current include a current value and an extraction time. The current value ranges from 200uA to 500uA, and the extraction time ranges from 1 min to 10 min.
在一个实施例中,第一生物传感层与第二生物传感层用于检测不同的生化物质,生化物质包括促黄体生成激素与促卵泡素。In one embodiment, the first biosensing layer and the second biosensing layer are used to detect different biochemical substances, and the biochemical substances include luteinizing hormone and follicle-stimulating hormone.
以上步骤的具体实现过程详见第一实施例的描述,在此不再赘述。The specific implementation process of the above steps can be found in the description of the first embodiment and will not be described again here.
本申请还提供一种计算机可读存储介质,所述计算机可读存储介质上存储计算机程序,所述计算机程序被处理器执行时实现如上实施例所述的生化物质检测方法的步骤。This application also provides a computer-readable storage medium. A computer program is stored on the computer-readable storage medium. When the computer program is executed by a processor, the steps of the biochemical substance detection method described in the above embodiments are implemented.
上述实施例仅例示性说明本申请的原理及其功效,而非用于限制本申请。任何熟悉此技术的人士皆可在不违背本申请的精神及范畴下,对上述实施例进行修饰或改变。因此,举凡所属技术领域中具有通常知识者在未脱离本申请所揭示的精神与技术思想下所完成的一切等效修饰或改变,仍应由本申请的权利要求所涵盖。The above embodiments only illustrate the principles and effects of the present application, but are not used to limit the present application. Anyone familiar with this technology can modify or change the above embodiments without departing from the spirit and scope of the present application. Therefore, all equivalent modifications or changes made by those with ordinary knowledge in the technical field without departing from the spirit and technical ideas disclosed in this application shall still be covered by the claims of this application.

Claims (10)

  1. 一种生化物质检测系统,其中,包括第一电极组、第二电极组与处理设备,所述处理设备与所述第一电极组、所述第二电极组连接;A biochemical substance detection system, which includes a first electrode group, a second electrode group and a processing device, the processing device is connected to the first electrode group and the second electrode group;
    所述第一电极组包括表面修饰有第一生物传感层的第一工作电极,所述第二电极组包括表面修饰有第二生物传感层的第二工作电极,所述第一生物传感层与所述第二生物传感层用于检测相同或不同的生化物质;The first electrode group includes a first working electrode with a first biosensing layer modified on its surface, and the second electrode group includes a second working electrode with a second biosensing layer modified on its surface. The sensing layer and the second biosensing layer are used to detect the same or different biochemical substances;
    所述处理设备被配置成用于:The processing equipment is configured for:
    通过所述第一电极组与所述第二电极组向检测区域施加脉冲电压,以对所述检测区域进行电穿孔处理;Apply a pulse voltage to the detection area through the first electrode group and the second electrode group to perform electroporation treatment on the detection area;
    通过所述第一电极组与所述第二电极组向所述检测区域施加恒电流,以使生化物质被提取至所述第一工作电极与所述第二工作电极所在区域的电解液;Apply a constant current to the detection area through the first electrode group and the second electrode group, so that biochemical substances are extracted into the electrolyte in the area where the first working electrode and the second working electrode are located;
    通过所述第一工作电极和/或所述第二工作电极对所述生化物质进行检测,确定所述生化物质的浓度。The biochemical substance is detected through the first working electrode and/or the second working electrode to determine the concentration of the biochemical substance.
  2. 根据权利要求1所述的生化物质检测系统,其中,所述第一电极组还包括第一对电极、第一参比电极、第一提取电极及第一柔性基底,所述第一工作电极、第一对电极、第一参比电极及第一提取电极设置在所述第一柔性基底上;The biochemical substance detection system according to claim 1, wherein the first electrode group further includes a first pair of electrodes, a first reference electrode, a first extraction electrode and a first flexible substrate, and the first working electrode, A first pair of electrodes, a first reference electrode and a first extraction electrode are provided on the first flexible substrate;
    所述第二电极组还包括第二对电极、第二参比电极、第二提取电极及第二柔性基底,所述第二工作电极、第二对电极、第二参比电极及第二提取电极设置在所述第二柔性基底上;The second electrode group also includes a second counter electrode, a second reference electrode, a second extraction electrode and a second flexible substrate. The second working electrode, the second counter electrode, the second reference electrode and the second extraction electrode Electrodes are provided on the second flexible substrate;
    其中,所述第一提取电极和所述第二提取电极用于向所述检测区域施加所述脉冲电压和所述恒电流。Wherein, the first extraction electrode and the second extraction electrode are used to apply the pulse voltage and the constant current to the detection area.
  3. 根据权利要求2所述的生化物质检测系统,其中,所述第一工作电极为片状电极,所述第一对电极与所述第一参比电极环绕所述第一工作电极的外周设置,所述第一提取电极环绕所述第一对电极的外周设置;The biochemical substance detection system according to claim 2, wherein the first working electrode is a sheet electrode, and the first pair of electrodes and the first reference electrode are arranged around the periphery of the first working electrode, The first extraction electrode is arranged around the periphery of the first pair of electrodes;
    所述第二工作电极为片状电极,所述第二对电极与所述第二参比电极环绕所述第二工作电极的外周,所述第二提取电极环绕所述第二对电极的外周。The second working electrode is a sheet electrode, the second pair of electrodes and the second reference electrode surround the outer periphery of the second working electrode, and the second extraction electrode surrounds the outer periphery of the second pair of electrodes. .
  4. 根据权利要求2所述的生化物质检测系统,其中,所述第一电极组设有第一支撑环,所述第一支撑环环绕所述第一电极组中的电极的外周设置;The biochemical substance detection system according to claim 2, wherein the first electrode group is provided with a first support ring, and the first support ring is arranged around the periphery of the electrodes in the first electrode group;
    所述第二电极组设有第二支撑环,所述第二支撑环环绕所述第二电极组中的电极的外周设置。The second electrode group is provided with a second support ring, and the second support ring is arranged around the periphery of the electrodes in the second electrode group.
  5. 如权利要求1所述的生化物质检测系统,其中,通过所述第一工作电极和/或所述第二工作电极对所述生化物质进行检测,确定所述生化物质的浓度时,所述处理设备被配置成用于:The biochemical substance detection system according to claim 1, wherein the biochemical substance is detected through the first working electrode and/or the second working electrode, and when the concentration of the biochemical substance is determined, the processing The device is configured for:
    采用差分脉冲伏安法,根据所述第一工作电极和/或所述第二工作电极产生的电化学信号,确定所述生化物质的浓度。Differential pulse voltammetry is used to determine the concentration of the biochemical substance based on the electrochemical signals generated by the first working electrode and/or the second working electrode.
  6. 根据权利要求1所述的生化物质检测系统,其中,用于施加所述脉冲电压的脉冲参数包括脉冲电压、脉宽、周期与脉冲个数,所述脉冲电压的范围为100V -500V,所述脉宽的范围为10 ms -50 ms,所述周期的范围为0 s -20 s,所述脉冲个数的范围为20个-200个。The biochemical substance detection system according to claim 1, wherein the pulse parameters for applying the pulse voltage include pulse voltage, pulse width, period and number of pulses, the range of the pulse voltage is 100V-500V, and the The pulse width ranges from 10 ms to 50 ms, the period ranges from 0 s to 20 s, and the pulse number ranges from 20 to 200.
  7. 根据权利要求1所述的生化物质检测系统,其中,用于施加所述恒电流的电流参数包括电流值与提取时间,所述电流值的范围为200μA -500μA,所述提取时间的范围为1 min -10 min。The biochemical substance detection system according to claim 1, wherein the current parameters used to apply the constant current include a current value and an extraction time, the current value ranges from 200 μA to 500 μA, and the extraction time ranges from 1 min -10 min.
  8. 根据权利要求1所述的生化物质检测系统,其中,所述第一生物传感层与所述第二生物传感层用于检测不同的生化物质,所述生化物质包括促黄体生成激素与促卵泡素。The biochemical substance detection system according to claim 1, wherein the first biosensing layer and the second biosensing layer are used to detect different biochemical substances, and the biochemical substances include luteinizing hormone and follicle-stimulating hormone. .
  9. 一种生化物质检测方法,其中,应用于如权利要求1-8中任一项所述的生化物质检测系统,所述方法包括:A biochemical substance detection method, which is applied to the biochemical substance detection system according to any one of claims 1 to 8, and the method includes:
    通过所述第一电极组与所述第二电极组向检测区域施加脉冲电压,以对所述检测区域进行电穿孔处理;Apply a pulse voltage to the detection area through the first electrode group and the second electrode group to perform electroporation treatment on the detection area;
    通过所述第一电极组与所述第二电极组向所述检测区域施加恒电流,以使生化物质被提取至所述第一工作电极与所述第二工作电极所在区域的电解液;Apply a constant current to the detection area through the first electrode group and the second electrode group, so that biochemical substances are extracted into the electrolyte in the area where the first working electrode and the second working electrode are located;
    通过所述第一工作电极和/或所述第二工作电极对所述生化物质进行检测,确定所述生化物质的浓度。The biochemical substance is detected through the first working electrode and/or the second working electrode to determine the concentration of the biochemical substance.
  10. 一种计算机可读存储介质,其中,所述计算机可读存储介质上存储计算机程序,所述计算机程序被处理器执行时实现如权利要求9所述的生化物质检测方法的步骤。A computer-readable storage medium, wherein a computer program is stored on the computer-readable storage medium, and when the computer program is executed by a processor, the steps of the biochemical substance detection method as claimed in claim 9 are implemented.
PCT/CN2023/106207 2023-07-06 2023-07-06 Biochemical substance measurement system and method, and storage medium WO2023179806A2 (en)

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