Method for detecting the amount of NO (nitric oxide) produced by a test subject and device implementing said method

Abstract

In the method of the present application, the physiological state of the subject can be tracked by studying the NO release profile of the subject, wherein said NO release profile is obtained by measuring the subject's skin by the sensing element during a predetermined sequence of activities.

Description

Technical Field

[0001] The present invention relates to a method and apparatus for detecting the physiological state and / or physiopathological state of a dead or living human, animal or plant subject, for example a subject. The present invention also relates to a standalone apparatus for measuring NO, the purpose of which is to determine the physiological or physiopathological state of a subject, for example to diagnose and / or prevent the occurrence of a pathology associated with the molecule and / or to monitor the efficacy of a treatment. Background Art

[0002] Nitric oxide is a gas that acts as an intercellular messenger. NO plays a crucial role in preventing the development and progression of certain cardiovascular diseases, neurodegenerative diseases, pulmonary hypertension, and tumors. Related cardiovascular diseases include hypercholesterolemia, hypertension, and diabetes. Most cardiovascular diseases (cerebrovascular, coronary, and lower limb ischemia) may be caused by endothelial dysfunction, which is associated with arteriosclerosis and can lead to thrombosis and ischemic lesions.

[0003] The cardioprotective effects of NO include, inter alia, regulating tonicity and vascular tone, inhibiting platelet aggregation, leukocyte adhesion, and the proliferation of smooth muscle fibroblasts. NO is also implicated in the development of bronchial inflammation. In particular, it has been measured that the concentration of NO in the air exhaled by subjects suffering from asthma is higher than that in the air exhaled by subjects who do not suffer from asthma. It has also been observed that, depending on the concentration of NO, NO is associated with the appearance or regression of tumors. It has also been observed that NO is associated with the pathology of Alzheimer's disease. All diseases affected by nitric oxide are long-term diseases, the annual cost of treating these long-term diseases is increasing year by year, and there is a need for tools to prevent and predict the onset of these diseases.

[0004] In physiology, nitric oxide is an excellent indicator of muscle growth and / or discomfort, making it an excellent tool for monitoring physical training in athletes and anyone engaging in physical activity. Therefore, by measuring nitric oxide production, injuries caused by overtraining can be avoided and / or NO uptake can be promoted, thereby promoting muscle growth and improving athletic performance. This applies to both humans and animals. In the context of cardiovascular disease, existing devices and preventive and prognostic tools have limitations. They can only indirectly measure NO in resting patients or directly, several hours after the pathological condition is observed. In all cases, measurements can only be performed in a clinical setting. Summary of the Invention

[0005] According to the present invention, a device is proposed that enables direct, continuous, and immediate measurement of NO in a biofluid, such as sweat, on the epidermis (e.g., skin) of a patient or mammalian subject, either in daily life or during a hospital visit, under varying pressure, humidity, and temperature conditions, optionally over several days and under all environmental conditions. This device makes it possible to detect and infer the development of physiological or physiological-pathological states, such as the risk of developing a pathology, or to monitor treatment.

[0006] A subject of the present invention is a method for detecting the amount of NO produced by a dead or living subject, in particular a human, animal or plant subject, in a predetermined sequence of activity states, characterized in that a study area of ​​the epidermis of the subject is selected and the production of NO dissolved in a biological fluid originating from the epidermis is directly and continuously tracked in this study area by a device formed by a first part, which is carried by the study area and is sealedly fixed to the study area, the first part being connected to a sensing element that detects NO by means of an electrochemical sensor, the method being further characterized in that the electrochemical sensor sends a signal due to the presence of an energy generator associated with the sensing element, and reading the signal of the electrochemical sensor makes it possible to achieve an ideal detection.

[0007] The expression "NO in biological fluids" is understood to mean that NO is dissolved in biological fluids.

[0008] The term "epidermis" is to be understood as the surface layer of plant tissue which forms the protective covering of the hollow parts of plants or the surface layer of the skin of humans and animals.

[0009] The expression "biological fluid originating from the epidermis" is understood to mean any fluid produced by a subject and excreted via or by the subject's epidermis. Such biological fluids are, for example, secretions in plants or sweat in humans and animals.

[0010] The expression "sealed" should be understood to mean that gases, liquids, and microorganisms (e.g., bacteria or viruses) outside the study area cannot enter the study area. The sealing of the contact between the first part and the study area ensures that the detected NO originates from the biological liquid produced in the study area and not from external flows.

[0011] The term "sequential" is understood to refer to a temporal order, ie a time interval.

[0012] The expression "predetermined activity state" should be understood to mean that the subject is in a state such as performing muscle movement, sleeping, sitting, running, still, or even dead.

[0013] According to one embodiment, the method makes it possible to detect at least one parameter related to a physiological state or a pathology.

[0014] According to one embodiment, the first portion comprises a fibrous body in order to transport the biological fluid from the investigation region to the sensing element by means of capillary forces.

[0015] According to one embodiment, the first portion further comprises a filter configured to filter the biological liquid at the inlet of the sensing element in order to avoid falsifying the detection of NO due to interfering elements contained in the biological liquid.

[0016] According to one embodiment, the filter is a eugenol type membrane.

[0017] According to some embodiments, the fiber body may be a woven material, as well as a non-woven material such as cotton.

[0018] In an alternative form of the method, at least one electrochemical sensor is used which provides a signal by electrochemical measurement using a biological fluid, in particular sweat or exudate, produced by the subject in the area of ​​study as an electrolyte between two working electrodes carried by an insulating planar support.

[0019] According to one embodiment, the insulating planar support comprises a material chosen from elastomers such as polydimethylsiloxane (PDMS), polyimide, epoxy and parylene.

[0020] In an alternative form of the method according to the invention, provision can be made for a reference electrode to be connected to both working electrodes.

[0021] According to one embodiment, the reference electrode is a silver chloride (AgCl) electrode.

[0022] It may also be provided that the sensing element comprises a plurality of similar electrochemical sensors, the signals of which are combined to improve the output signal.

[0023] Provision can be made for the pattern of the electrodes relative to their support to follow a Hilbert curve in order to improve the power of the output signal per unit area of ​​said support.

[0024] According to one embodiment, the pattern of the electrodes relative to their support may follow another type of curve selected from the Peano curve, the Sierpiński curve, the Moore curve and the Lebesgue curve, also in order to improve the intensity of the output signal per unit area of ​​the support.

[0025] Provision may be made that the measurement is carried out in line with a hole provided in the planar support, the hole being in line with the conductive pattern of the electrode.

[0026] In an advantageous embodiment, the electrode consists of a metal deposit, in particular a deposit of silver (Ag), gold (Au), platinum (Pt) and platinum black, or a graphene deposit doped with silver (Ag) or gold (Au) nanoparticles, said nanoparticles being functionalized with a NO binder, in particular guanylate cyclase or porphyrin.

[0027] According to one embodiment, the gold metal deposits are produced in clusters or by following a precise pattern, for example a hexagonal pattern.

[0028] For one embodiment of the method according to the invention, it can be provided that the device also includes a second part located above the first part, which second part contains electronic components for receiving the raw measurement values ​​from the electrochemical sensor, converting the raw measurement values ​​into NO concentration and ensuring the transmission of the signal using parameters that may be related to the environment.

[0029] In the method according to the invention, provision can be made for the device to perform and transmit measurements at a frequency that is dependent on the activity state of the object, which is tracked by the gyroscope and / or accelerometer of the second part of the device.

[0030] According to one embodiment, the apparatus comprises a geo-location module.

[0031] The present invention also relates to a detection device for detecting the amount of NO produced by a subject in a sequence of active states, the device comprising a first part and a second part, the first part being intended to be carried by a study area of ​​the subject's epidermis and sealingly fixed to the study area so as to directly and continuously track the production of NO in biological fluid originating from the epidermis, the first part being connected to a sensing element that detects NO through an electrochemical sensor, the second part being configured to send a signal, wherein the signal can be sent due to an energy generator associated with the sensing element, and ideal detection can be achieved by reading the signal.

[0032] Provision may be made for the sensing element to provide a signal obtained by electrochemical measurement using a biological fluid produced by the subject in the investigation area as an electrolyte between two working electrodes carried by an insulating planar support.

[0033] In the above-mentioned alternative, provision can be made for a reference electrode to be connected to the two working electrodes.

[0034] According to one embodiment, the insulating planar support comprises at least one microchannel to guide the biological fluid to the electrochemical sensor.

[0035] It may be provided that the sensing element comprises a plurality of similar electrochemical sensors, the signals of which are combined in order to improve the output signal.

[0036] According to one embodiment, the sensing element comprises a plurality of electrochemical sensors distributed in a plurality of sensing cells, and each sensing cell is configured to detect at least one chemical substance.The sensing element can then detect several different chemical substances.

[0037] According to one embodiment, the insulating planar support comprises a plurality of microchannels, and each channel comprises a sensing unit.

[0038] In the device according to the invention, provision can be made for the pattern of the electrodes relative to their support to follow a Hilbert curve in order to improve the intensity of the output signal per unit area of ​​the support.

[0039] In such an arrangement, the measurements are performed in line with a hole provided in the planar support, which hole is in line with the conductive pattern of the electrode.

[0040] In the device according to the invention, it can be provided that the working electrode consists of a metal deposit, in particular a deposit of silver (Ag), gold (Au), platinum (Pt) and platinum black, or a graphene deposit doped with silver (Ag) or gold (Au) nanoparticles, the nanoparticles being functionalized with a binder for NO, in particular guanylate cyclase or porphyrin.

[0041] According to one embodiment, the first portion comprises a fibrous body in order to transport the biological fluid from the investigation region to the sensing element by means of capillary forces.

[0042] According to one embodiment, the first portion further comprises a filter configured to filter the biological liquid at the inlet of the sensing element in order to avoid falsifying the detection of NO due to interfering elements contained in the biological liquid.

[0043] In the device according to the invention, provision can be made for the second part to be located above the first part, the second part containing electronic components for receiving the raw measured values ​​from the electrochemical sensor, converting the raw measured values ​​into NO concentrations, and ensuring signal transmission, possibly with parameters related to the environment.

[0044] In the device according to the invention, provision can be made for the device to perform and transmit measurements at a frequency that is dependent on the activity state of the object, which is tracked by the gyroscope and / or accelerometer of the second part of the device.

[0045] According to one embodiment, the apparatus comprises a geo-location module. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] In order to make the subject matter of the invention more easily understood, a description will be given below of one embodiment thereof by way of a purely illustrative and non-limiting example shown in the accompanying drawings, in which

[0047] - Figure 1 The external overall view of the detection device according to the present invention is shown in a perspective view;

[0048] - Figure 2 shows an overall view of an object on which a device according to the invention has been placed;

[0049] - Figure 3 Shown Figure 1 Exploded view of the device;

[0050] - Figure 4 Shown with Figure 3 A block diagram corresponding to the operation of the device;

[0051] - Figure 5 Shown is a top view of a planar support with two electrodes positioned according to the Hilbert curve.

[0052] - Figure 6 Figures obtained from healthy subjects are shown. Figure 2 The device according to the present invention is shown;

[0053] - Figure 7 A first arrangement of a fiber body and sensing elements of the device is schematically shown;

[0054] - Figure 8 schematically shows a second arrangement of the fiber body and sensing elements of the device;

[0055] - Figure 9 schematically illustrates an electrochemical sensor comprising a sensing element of three electrodes according to a first embodiment;

[0056] - Figure 10 schematically illustrates an electrochemical sensor comprising a sensing element of three electrodes according to a second embodiment;

[0057] - Figure 11 is a functional schematic diagram of a micro-hydraulic circuit arranged in a sensing element;

[0058] - Figure 12 is a cross-sectional view of a sensing element according to one embodiment. DETAILED DESCRIPTION

[0059] With reference to the accompanying drawings, it can be seen that the detection device according to the present invention is always indicated by 1; the detection device is intended to quantitatively measure NO in healthy human subjects. In the example described, the subject performs physical exercise by using a bicycle with a power of 160W. Figure 6 As shown, the time sequence from the detection of NO at the beginning of the test (point 11) to the end of the test (point 12) is about 500 seconds. Figure 1 , the device 1 is generally in the form of a self-adhesive portion, which in the example takes the form of a self-adhesive pad, which can be placed directly on the skin of a subject. In one embodiment not shown, the self-adhesive portion is a self-adhesive dressing.

[0060] The device according to the present invention comprises a fastening base 3 made of a biocompatible, adhesive, flexible material; this base ensures that the entire device is fixed to the skin. A central portion 4a of the base 3 is a circular recess, within which the first portion of the device is located, thereby allowing the tracking of NO production in the target area of ​​the subject's skin. Thus, the circular recess 4a enables the first portion of the measuring device to be placed directly on the subject's skin 2. Recess 4a may also have another shape, selected from, for example, an oval, triangular, rectangular, square, or polygonal shape.

[0061] The first part comprises a fiber body 4 which is attached to a sensing element 5, such as Figure 7 As shown, the sensing element is suspended, or as Figure 8 As shown, the sensing element is wrapped to form the base of the stack. The fibrous body has the function of transporting the sweat produced in the area of ​​interest to the sensing element 5, where it detects the nitric oxide dissolved therein, and then expelling the sweat once the measurement has been made.

[0062] A filter 29 may be optionally arranged between the fiber body 4 and the inlet of the sensing element 5. The function of the filter 29 is to filter the sweat to prevent certain elements naturally contained therein from interfering with the measurement of NO dissolved in the sweat. These interfering elements are, for example, peroxynitrite (ONOO - ) or hydrogen peroxide (H2O2).

[0063] The sensing element 5 detects NO by means of one or more electrochemical sensors 14, which will be defined below. The sensor sends its information to a converter 6a, which itself provides a processor 6b, which is powered by an energy generator 6d associated with the sensing element 5. The processor 6b provides a radio communication system 6c, which sends the information to a network capable of converting the information into a graph, such as Figure 6 An instrumentation of the type shown in FIG.

[0064] exist Figure 6The portion of the graph that constitutes the measurement of NO produced by the subject during effort is the portion between points 11 and 12 of the graph. Figure 6 The integral of the graph between points 11 and 12 corresponds to a parameter. The value of this parameter can be linked to pathologies such as arteriosclerosis. It can also be used to manage physiological functions, such as monitoring alanine bioavailability. Specifically, the natural precursor of NO in living organisms is the amino acid alanine. The body only produces NO in response to exertion within the limits of its alanine reserves. Therefore, this device can also predict the moment when a subject will no longer be able to control vasodilation, thereby predicting the risk of injury.

[0065] implement Figure 4 All components of the various functions shown are assembled in an embedded electronic system, generally designated 6. Components 4, 5 and 6 form a stack fixed to the skin of the subject by a flexible and watertight envelope of the silicone type, generally designated 7.

[0066] The embedded electronic system of component 6 performs the functions of controlling the components of sensing element 5. The embedded electronic system of component 6 also includes gyroscope and accelerometer units to understand the subject's orientation and movement, as well as the start and end of the subject's activity sequence, and a temperature sensor for measuring skin temperature. Knowing the skin temperature is useful in order to be able to correlate temperature with blood vessel dilation.

[0067] The sensing element of the described examples is electrochemical. Figure 5 The sensing element shown in FIG. 1 comprises an electrochemical sensor 14 and an insulating planar support 10 made of polyimide. The electrochemical sensor comprises two electrodes 8 and 9, which are located on one side of the insulating planar support 10 and between which is the sweat produced by the subject in the area of ​​interest, i.e. in line with the stack 4, 5, 6. Figure 5 In the sensing element shown, four identical units can be seen, each capable of acquiring NO measurements. Advantageously, installing several sensing units allows obtaining a skin map of NO production within the covered area. Figure 7 , the arrangement of the fiber body 4 and the sensing element 5 is Figure 3 Different. An insulating planar support member 10 is placed directly on the skin 2. The side of the support member 10 provided with the electrochemical sensor is on the opposite side of the side in contact with the skin 2. The fibrous body 4 has a portion that contacts the skin and a portion that covers one side of the support member, and the portion covering the support member includes the electrochemical sensor. In other words, the filter 4 spans the skin and the electrochemical sensor. In this embodiment, the fibrous body comprises cotton or a nonwoven material.

[0068] refer to Figure 8, shows a second arrangement of the fiber body 4 and the sensing element 5. The fiber body 4 sandwiches the sensing element. As a result, a portion of the fiber body 4 is placed against the skin. Next, the sensing element 5 is placed on the portion of the fiber body that is close to the skin. The portion of the fiber body 4 not placed against the skin is folded over the sensing element 5, thereby covering the sensor.

[0069] According to the first embodiment, Figure 9 As schematically shown in FIG, electrochemical sensor 14 includes three electrodes: a reference electrode 20, a working electrode 21, and an auxiliary electrode 22. Reference electrode 20 is a silver chloride (AgCl) electrode, auxiliary electrode 22 is a platinum (Pt) electrode, and working electrode 21 is a platinum black electrode. Working electrode 21 is disk-shaped. The disk is partially surrounded by the reference electrode and the auxiliary electrode, which are opposite each other. The electrochemical sensor has dimensions on the order of millimeters.

[0070] according to Figure 10 In the second embodiment shown, the electronic sensor includes a reference electrode 20, a working electrode 21, and an auxiliary electrode 22. The reference electrode 20 is a silver chloride (AgCl) electrode, the auxiliary electrode 22 is a platinum (Pt) electrode, and the working electrode 21 is a platinum black electrode. The working electrode 21 is disk-shaped. The disk is partially surrounded by the reference and auxiliary electrodes. The electrodes are concentrically arranged: the working electrode 21 is partially surrounded by the reference electrode 20, which itself is surrounded by the auxiliary electrode 22. The dimensions of the electrochemical sensor are approximately millimeters.

[0071] Figure 9 or Figure 10 Electrochemical sensors can be used for Figure 11 The micro-hydraulic circuit is schematically shown in FIG.

[0072] exist Figure 11 In the example shown, the fiber body 4 absorbs biological fluid, here sweat, and transports it to three microchannels 15 marked on the planar support 10. These microchannels 15 will each transport the sweat to a sensing unit 16, 17, 18. In the example shown, there is one sensing unit for each microchannel 15. Sensing unit 16 will detect nitric oxide, sensing unit 17 will detect nitrite contained in sweat, and sensing unit 18 will detect hydrogen peroxide contained in sweat. Nitrite is mainly converted into nitric oxide by superoxide (O2 -· ) is produced inside cells by the reaction between NO2- and nitric oxide. Therefore, by detecting NO2-, the NO concentration can be better measured.

[0073] Thus, each sensing cell is used for the detection of a chemical substance. Each sensing cell is powered so that each sensing cell is at an applied potential in order to perform a fixed measurement. Sensing cell 18 is at the redox potential of hydrogen peroxide (oxidizing substance) in order to detect hydrogen peroxide. Processing of the data from sensing cell 18 will give the amount of H2O2. Sensing cell 16 is at the redox potential of NO (oxidizing substance) in order to detect NO. Since the redox potential of H2O2 is lower than the redox potential of NO, sensing cell 16 detects H2O2 and NO. Processing of the data from sensing cell 16 will give the total amount of H2O2 and NO. Sensing cell 17 is at the redox potential of nitrite (oxidizing substance) in order to detect NO2 - When NO2 - When the redox potential of H2O2 is higher than that of NO, unit 17 detects H2O2, NO and NO2 - The data from the sensing unit 17 is processed to give H2O2, NO and NO2 - By subsequent processing of the data generated by the sensing units 16, 17, 18, the amount of each chemical substance, namely NO, H2O2 and NO2 can be determined by the difference. - amount.

[0074] Alternatively, a pulse method could be used and then each sensing unit would be able to detect each species. After processing the data, the amount of each substance present would be able to be determined.

[0075] refer to Figure 12 , the sensing element 5 includes three microchannels 30, 31, 32 marked on the thickness of the insulating planar support 10. Sensing units 16, 17 and 18 are placed on each bottom wall of the microchannels 30, 31. Sensing unit 16 is configured to detect nitric oxide, sensing unit 17 is configured to detect nitrite contained in sweat, and sensing unit 18 is configured to detect hydrogen peroxide contained in sweat. Each sensing unit 16, 17, 18 includes three sensors 14. A filter 29 is placed on top of the insulating planar support. The filter covers the microchannels. Finally, the fiber body 4 is placed on the filter 29.

[0076] Fibrous body 4 absorbs biological fluid, in this case sweat, and transports it to three microchannels 30, 31, and 32 through capillary action. When the sweat discharged from fibrous body 4 reaches the level of the microchannels, it is filtered by filter 29 to remove certain interfering elements, and then transported by microchannels 30, 31, and 32 to at least sensing units 16, 17, and 18. The sensor of sensing unit 16 then detects NO, the sensor of sensing unit 17 detects nitrite, and the sensor of sensing unit 18 detects hydrogen peroxide.

[0077] In an embodiment not shown, when the sensing element comprises a plurality of sensing cells, at least one of which is dedicated to detecting a chemical substance other than NO, such as hydrogen peroxide, the filter 29 may be omitted.

[0078] The current intensities obtainable with the device according to the invention are between the picoampere and milliampere range.

[0079] Although the present invention has been described in conjunction with a number of specific embodiments, it is very obvious that the present invention is not limited thereto in any way and comprises all technical equivalents of the described means and their combinations, provided that they all fall within the scope of the present invention.

[0080] Use of the verb "comprise", "have" or "include" and their conjugated forms does not exclude the presence of elements or steps other than those listed in a claim.

[0081] In the claims, any reference signs placed between parentheses shall not be construed as limiting the claim.

Claims

1. A method for detecting the amount of NO produced by a subject under a predetermined sequence of activity states for non-diagnostic purposes, wherein: a study area of ​​the epidermis (2) of the subject, and the production of NO dissolved in a biological fluid originating from the epidermis is tracked directly and continuously in the study area by means of a device formed by a first part (4), which is located on the study area and is fixed sealingly thereto, the first part (4) being connected to a sensing element (5), wherein the sensing element comprises a plurality of electrochemical sensors distributed in a plurality of sensing cells, wherein each sensing cell is configured to detect at least one chemical substance, wherein a first sensing cell in the plurality of sensing cells is configured to detect NO via a first electrochemical sensor in the plurality of electrochemical sensors, and a second sensing cell in the plurality of sensing cells is configured to detect NO2 via a second electrochemical sensor in the plurality of electrochemical sensors. - , wherein the first electrochemical sensor sends a first signal and the second electrochemical sensor sends a second signal due to the presence of an energy generator associated with the sensing element (5), and reading the signals makes it possible to perform a detection, wherein the first and second electrochemical sensors provide signals by electrochemical measurements using a biological fluid produced by the subject in the investigation area, the biological fluid acting as an electrolyte between two working electrodes located on an insulating planar support, wherein the working electrode of at least one of the first and second electrochemical sensors is composed of a metal deposit of platinum black, The device further comprises a converter, a processor and an instrument, wherein the converter powered by the energy generator converts the signal sent by the first electrochemical sensor and provides the processor, the processor provides a radio communication system, the radio communication system sends information to the instrument which converts the information into a curve graph, the method further comprising detecting at least one parameter associated with the physiological state, wherein the parameter corresponds to the integral of the curve graph between a first point and a second point, wherein the first point is the point at which the test starts and the second point is the point at which the test ends.

2. The method according to claim 1, wherein A reference electrode is connected to the two working electrodes (8, 9).

3. The method according to any one of claims 1 to 2, wherein The measurements are performed in line with a hole (13) provided in the planar support, which hole is in line with the conductive pattern of the electrodes (8, 9).

4. The method according to any one of claims 1 to 2, wherein The device performs and sends measurements at a frequency related to the activity state of the object, which state is tracked by a gyroscope and / or accelerometer module of the second part (6) of the device.

5. The method according to claim 1, wherein The plurality of sensing units include a third sensing unit and the plurality of electrochemical sensors include a third electrochemical sensor, and wherein the third sensing unit is configured to detect H2O2 through the third electrochemical sensor of the plurality of electrochemical sensors, wherein the insulating planar support comprises a plurality of microchannels to guide the biological fluid to the electrochemical sensor and wherein each microchannel comprises a sensing cell, a first microchannel of the plurality of microchannels comprises the first sensing cell, a second microchannel of the plurality of microchannels comprises the second sensing cell and a third microchannel of the plurality of microchannels comprises the third sensing cell, The first sensing unit is at the oxidation-reduction potential of NO to detect NO, and the second sensing unit is at the NO2 - Redox potential to detect NO2 - and the third sensing unit is at the oxidation-reduction potential of H2O2 so as to detect H2O2, The data from the second sensing unit is processed to give H2O2, NO and NO2 - The total amount of NO, H2O2 and NO2 can be determined by the difference by further subsequent processing of the data generated by the first sensing unit, the second sensing unit and the third sensing unit. - amount.

6. A detection device (1) for detecting the amount of NO produced by a subject under a test of a predetermined sequence of activity states, the device comprising a first part (4) intended to be located on a study area of ​​the epidermis (2) of the subject and sealed to the study area so as to directly and continuously track the production of NO in a biological fluid originating from the epidermis, the first part being connected to a sensing element (5), wherein the sensing element comprises a plurality of electrochemical sensors distributed in a plurality of sensing cells, wherein each sensing cell is configured to detect at least one chemical substance, wherein a first sensing cell of the plurality of sensing cells is configured to detect NO via a first electrochemical sensor (14) of the plurality of electrochemical sensors, and a second sensing unit among the plurality of sensing units is configured to detect NO2 through a second electrochemical sensor among the plurality of electrochemical sensors. - , The device comprises a second part (6) configured to transmit a first signal and a second signal, wherein the first signal and the second signal are transmitted due to an energy generator (5d) associated with the sensing element (5), and by reading the first signal and the second signal it is possible to detect the amount of NO generated during the sequence and the amount of NO2 generated during the sequence - The amount, wherein the sensing element provides the first and second signals, the first and second signals being obtained by electrochemical measurements of the first of the plurality of electrochemical sensors and the second of the plurality of electrochemical sensors using a biological fluid produced by a subject within the investigation area as an electrolyte between two working electrodes located on an insulating planar support, wherein the working electrode of at least one of the first and second electrochemical sensors is composed of a metal deposit of platinum black, The detection device further includes a converter, a processor and an instrument, wherein the converter powered by the energy generator is configured to convert the signal sent by the first electrochemical sensor and provide the processor, the processor is configured to provide a radio communication system, the radio communication system is configured to send information to the instrument, the instrument is configured to convert the information into a curve graph, and the detection device is configured to detect at least one parameter related to a physiological state or pathology, wherein the parameter corresponds to the integral of the curve graph between a first point and a second point, wherein the first point is the point at which the test starts and the second point is the point at which the test ends.

7. The device according to claim 6, wherein A reference electrode is connected to the two working electrodes.

8. The device according to any one of claims 6 and 7, wherein The insulating planar support (10) comprises at least one microchannel (15, 30, 31, 33) for guiding biological fluid to the electrochemical sensor (14).

9. The device according to any one of claims 6 and 7, wherein The pattern of the working electrodes (8, 9) relative to their support (10) follows a Hilbert curve in order to improve the intensity of the output signal per unit area of ​​the support.

10. The device according to any one of claims 6 and 7, wherein The measurements are performed in correspondence with a hole provided in the planar support (10) which is in correspondence with the conductive pattern of the electrodes (8, 9).

11. The device according to claim 8, wherein The insulating planar support comprises a plurality of microchannels (30, 31, 32), and each microchannel comprises a sensing unit.

12. The device according to any one of claims 6 and 7, wherein The first portion comprises a fiber body for transporting the biological fluid from the investigation region to the sensing element by means of capillary forces.

13. The device according to claim 12, wherein The first part also comprises a filter (29) configured to filter the biological fluid at the inlet of the sensing element in order to avoid falsifying the detection of NO due to interfering elements contained in the biological fluid.

14. The device according to any one of claims 6 and 7, wherein The second part is located above the first part (4) and contains electronic components for receiving raw measurement values ​​from the electrochemical sensor, for converting the raw measurement values ​​into NO concentration and ensuring the transmission of the signal with possible parameters related to the environment.

15. The device according to any one of claims 6 and 7, wherein The device includes a gyroscope and / or accelerometer module to detect the activity state of the object, and the device is configured to take and transmit measurements at a frequency related to the activity state of the object.

16. The device according to claim 6, wherein The plurality of sensing cells include a third sensing cell and the plurality of electrochemical sensors include a third electrochemical sensor, and wherein the third sensing cell is configured to detect H 2 O 2 through the third electrochemical sensor of the plurality of electrochemical sensors.

17. The device according to claim 6 or 16, wherein Each of the plurality of sensing cells includes three electrochemical sensors.

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