• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
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    • 专利摘要:
    The present invention consists of an electrode device for the detection of ascorbic acid, with three electrodes, all the electrodes obtained by screen printing, as well as its manufacturing method and its use in the mentioned detection. (Machine-translation by Google Translate, not legally binding)
    公开号:ES2660516A1
    申请号:ES201631238
    申请日:2016-09-22
    公开日:2018-03-22
    发明作者:Maria Julia ARCOS MARTÍNEZ；Maria Asunción ALONSO LOMILLO；Olga DOMÍNGUEZ RENEDO；Abraham BOTÍN SALDAÑA
    申请人:Universidad de Burgos；
    IPC主号:
    专利说明:

    ELECTRONIC DEVICE FOR DETECTION OF ASCORBIC ACID, MANUFACTURING PROCEDURE AND USE OF SUCH DEVICEDESCRIPTION 5
    OBJECT OF THE INVENTION

    The present invention is framed in the field of electrochemistry applied to chemical analysis. 10

     Thus, the present invention relates to a device consisting of an electrode system for the detection of ascorbic acid, with three electrodes, all electrodes obtained by screen printing, as well as its manufacturing process and conditions of use in the aforementioned detection. .
    BACKGROUND OF THE INVENTION
     twenty
    The development of reproducible and sensitive devices for electrochemical detection of anti-oxidant ascorbic acid, which helps in detoxification and improves iron absorption in the body (PJ O'Connell, C. Gormally, M. Pravda, GG Guilbault, Development of an 25 amperometric L-ascorbic acid (Vitamin C) sensor based on electropolymerised aniline for pharmaceutical and food analysis, Analytica Chimica Acta, 431 (2001) 239), is still being studied today, as evidenced by the more than 10,000 scientific publications which can be found in the different databases. It has also been shown that the concentration of ascorbic acid in biological fluids can be used to assess oxidative stress in human metabolism, which has been related in turn to liver disease, cancer or diabetes (A. Ambrosi, A. Morrin, MR Smyth, AJ Killard, The application of conducting polymer nanopar-ticle electrodes to the sensing of ascorbic acid, Analytical Chimica Acta, 609 (2008) 37).
     5
    Traditionally, spectroscopic, chromatographic and electrochemical methods have been used for the detection of ascorbic acid. The latter techniques are characterized by their simplicity, speed and portability, which also makes possible the detection in situ of the analyte of interest (CA Fuenmayor, S. Benedetti, A. Pellicanò, MS Cosio, S. Mannino, Direct In Situ Determination of Ascorbic Acid in Fruits by Screen-Printed Carbon Elec-trodes Modified with Nylon-6 Nanofibers, Electro-analysis, 26 (2014) 704). However, in the detection of ascorbic acid especially in complex matrices such as food products or biological samples, relatively high working potentials are used for electrochemical oxidation to occur, which is associated with poor selectivity, and electro-20 deactivation. Working principally caused by the accumulation of reaction products on the electrode surface, resulting in a progressive deterioration of electrode sensitivity (PR Roy, MS Saha, T. Okajima, T. Ohsaka, Electrocatalytic oxidation of ascorbic acid by 25 [Fe (CN) 6] 3− / 4− redox couple electrostatically trapped in cationic N, N-dimethylaniline polymer film electropolyme-rized on diamond electrode, Electrochimica Acta, 51 (2006) 4447) (CA Fuenmayor, S. Benedetti, A. Pelli-canò, MS Cosio, S. Mannino, Direct In Situ Determination-30 tion of Ascorbic Acid in Fruits by Screen-Printed Carbon Electrodes Modified with Nylon-6 Nanofibers, Electro-an alysis, 26 (2014) 704.
    These drawbacks have been attempted to minimize by using simple disposable devices manufactured by screen printing. bles manufactured by screen printing. bles manufactured by screen printing. bles manufactured by screen printing. bles manufactured by screen printing. bles manufactured by screen printing. bles manufactured by screen printing. bles manufactured by screen printing. bles manufactured by screen printing. bles manufactured by screen printing.

    Screen printing is a direct printing method, also called penetration printing. The deposition of inks is done layer by layer on a substrate. 25 The quality of the chemical sensors thus manufactured depends, to a large extent, on the materials used.
    By means of screen-printed electrode technology, the miniaturization of the sensors is possible, offering the advantage of being versatile, they can be manufactured with different electrode configurations and with different inks, and have a low cost and can thus lend themselves to mass production Disposable electrodes can also be used for in situ analysis.
    The differential pulse voltammetry technique consists in the application of a potential sweep to the electrode system, on which it periodically superimposes a series of potential impulses. The signal that is recorded is the difference in intensity after and before applying the pulse against the base potential. This differential current measurement eliminates capacitive intensities and background noise, increasing the sensitivity of the analysis. 10

    In screen-printed electrode systems, the reference electrode is usually made up of a conductive paste based on Ag, so it does not act as a true reference but as a pseudo-reference. fifteen

    The electrochemical methods indicated above for the determination of ascorbic acid are poorly selective and do not allow quantification of this compound in complex matrices. The present invention solves the aforementioned disadvantages, and proposes a device that works at a lower potential avoiding interference from other compounds in complex samples. The device has great simplicity, low cost and short analysis time, compared to the rest of the 25 techniques commonly used for its determination, so it is a great advance in its use for this determination in different types of samples such as samples Biological
     30 DESCRIPTION OF THE INVENTION

    The object of the invention is an electrode device, its manufacturing process and use thereof for the detection of ascorbic acid. The problem to be solved is the selective determination of ascorbic acid with simpler techniques, with low cost and in a relatively short analysis time compared to known techniques for the determination of ascorbic acid.

    In view of the foregoing, in one aspect, the present invention relates to an electrode device for the detection of ascorbic acid in 10 different types of liquid samples, obtained by screen printing characterized in that it comprises three electrodes, each electrode. it consists of a contact, a section and an active area, being a working electrode, an auxiliary or counter electrode, and a reference electrode, constituted two electrodes by screen printing of an ink of conductive materials on a plastic sheet of polyester. The conductive tracks constituted by the contacts also called terminals (1.3, 2.3, 3.3) and the sections (1.2, 2.2, 3.2) have been printed with an ink at 20 Ag base, the active areas of the electrodes that constitute the counter electrode ( 3.1) and the working electrode (2.1), have been printed with a carbon conductive ink and the reference one with an Ag / AgCl ink in the active area. 25

    In another aspect, the invention relates to the manufacturing method of the electrode device described above, whose spatial arrangement allows rapid "in situ" analysis of the sample by electrochemical techniques.

    The manufacturing process, screen printing of the device is carried out with the help of a series of screens, also called molds or screen printing patterns 35, which allow defining the different electrodes on the same support. The process of silkscreen construction basically involves several stages consisting of the sequential deposition of the different inks and their subsequent curing. 5

    Four different screens were designed for the manufacture of these devices, corresponding to each level of ink deposition, as follows:
      10
    - The first screen defines the conductive tracks, constituted by the contacts (1.3, 2.3, 3.3) and the sections (1.2, 2.2, 3.2) printed with an ink based on Ag.
     fifteen
    - The second screen defines the shape and position of the reference electrode.

    - The third screen defines the active area of the counter electrode (3.1) and the active area of the working electrode (2.1)

    - The fourth screen is designed for the deposition of an insulating material to prevent contact between the solution and the conductive tracks, constituted by the contacts and the sections.

    The set constitutes a miniaturized sensor whose geometry is optimal for the analysis of small real liquid samples. The device can be introduced into an electrochemical cell in which the sample is in solution.

    To put the device into operation, the contacts or terminals of each of the three electrodes (1.3, 2.3, 3.3) are connected to the output terminals of a potentiostat through electrical connections.

    The present invention relates in another aspect 5 to the use of the aforementioned device comprising the following steps:

    - Modification of the working electrode with gold na-noparticles. The deposition of the na-10 gold noparticles (AuNPs) on the working electrode is carried out electrochemically by deposing a drop of 100 µL of 0.1 mM solution of HAuCl4 prepared in 0.5SO H2SO4, and applying a potenti- 15 cial constant +0.18 V vs Ag / AgCl SPE for 50 seconds.

    - Introduction of the device in the electro-trochemical cell. twenty

    - Adding 5 mL of the sample in the presence of a 100 mM buffer solution of KH2PO4 and 100 mM KCl of pH = 7.
     25
    - Application of a potential of +0.2 V vs Ag / AgClSPE.

    - Carrying out the amperometric measurements recording the intensity in the different screen-printed positive dis-30s.
    DESCRIPTION OF THE FIGURES

    The present specification is complemented, with a set of figures, illustrative of the preferred and never limiting example of the invention.

    Figure 1 shows a scheme of the electrode reaction. 5

    Figure 2 represents an elevation view of the electrodes without insulation.

    Figure 3 represents an elevation view of the 10 insulated electrodes.

    Figure 4 shows a type amperogram obtained in the determination of ascorbic acid.
     fifteen DETAILED EXHIBITION OF THE INVENTION

    In the embodiment detailed herein, the invention relates to an electrode device for the detection of ascorbic acid, with three electrodes (1, 2, 3), all electrodes obtained by screen printing, as well as their manufacturing process, and its use in the cited detection.

    As the screen printing technique involves the use of 25 concrete materials and gives rise to the specific geometric configuration of the device, the exposure of said manufacturing technique also serves as an explanation of the configuration of the device, both of which can be linked, configuration and technique or manufacturing process 30, during the present detailed exposition of the invention.

    The reaction mechanism proceeds according to the scheme shown in Figure 1. Ascorbic acid 35 is oxidized to dehydroascorbic acid, the electrons being assigned to the electrode in which the reaction takes place.

    The invention relates to the device, method of manufacturing and using a disposable three-electrode device (1, 2, 3), screen printed on a polyester support, in particular ethylene polyterephthalate (PET), whose spatial arrangement allows the rapid “in situ” analysis of small sample volumes by electrochemical techniques. 10

    The device consists of three serigraph electrodes (1, 2, 3): reference electrode (1), working electrode (2) and auxiliary electrode (3) or counter electrode. fifteen

    The electrodes have three distinct zones or parts: the contacts (1.3, 2.3, 3.3) or terminals, for connection to a known potentiostat; the active areas (1.1, 2.1, 3.1), in direct contact with the sample to be analyzed and the sections (1.2, 2.2, 3.2), as a union between contacts (1.3, 2.3, 3.3) and the active areas (1.1, 2.1 , 3.1). Being the set of contacts (1.3, 2.3, 3.3) and the sections (1.2, 2.2, 3.2) the conductive routes.
     25
    The process of manufacturing, screen printing, of the device is carried out with the help of a series of screens, molds or screen printing patterns in which, on a porous surface, the scheme of the motif to be printed that will define the device appears. 30

    - The first screen or pattern is used to form, conducting with conductive Ag commercial paste the conductive paths constituted by the contacts (1,3), (2,3), (3,3) and the sections (1,2) , (2,2), (3,2) of the electrodes (1, 2, 3). That is, the electrical conductive base of the device is created. After application this layer should be cured for 30 minutes at 120 ° C.
     5
    - The second screen or pattern is designed to obtain the active area (1.1) of the reference electrode (1). For its formation, using this second screen, a commercial product based on Ag / AgCl is printed on the conductive base in the area of the active area 10 of said electrode, which is subsequently subjected to a healing process under the same conditions, 20 minutes at 120 ° C.

    - The third screen or pattern is designed to print together the active areas of the counter-electrode (3.1) and the working electrode (2.1) using carbon ink.

    - The fourth screen or pattern is designed to print the insulating layer that covers the device, freeing the contacts (1.3, 2.3, 3.3) and active areas (1.1, 2.1, 3.1) of the electrodes, using an insulating ink. The correct formation of this insulating layer requires healing at 80 ° C for 30 minutes. 25

    In this embodiment, the active area of the reference electrode is of rectangular geometry, the circular one and the counter electrode has an arcuate envelope shape of the other two electrodes. The best 30 dimensions of the active area (1.1) of the device reference electrode correspond to 2 mm wide by 4 mm long.

    For the working electrode (2) the best proven dimension of its active area (2.1) corresponds to a circle of 4.5 mm in diameter.

    To put the device into operation, the upper conductive parts (1.3, 2.3, 3.3) of each of the electrodes (1, 2, 3) are connected to the output terminals of a potentiostat through electrical connections .
     10
    100 microliters of a gold salt are deposited in the electrode device, and it is subjected to a 0.18 V potential to allow the modification of the surface of the working electrode with gold nanoparticles. Next, the electrode device 15 is immersed in a cell with the solution to be analyzed and according to the technique selected to carry out the amperometric determination, the electrode device is subjected to a potential difference of 0.2 V, registering the intensity that originates in the circuit due to the electrochemical reactions that the potential disturbance causes, in the analyte ascorbic acid.

    To carry out the determination of a sample ascorbic acid problem, the ascorbic acid sample is added to the cell where the electrode device was introduced in the presence of a phosphate buffer solution of pH = 7.0 and subjected to dissolution at a constant stirring speed. Subsequently, successive additions of standard solutions of ascorbic acid are made and the intensities recorded after each addition are measured.

    The corresponding calibration line is obtained and the concentration of ascorbic acid in the test sample is calculated by extrapolation, according to the standard addition procedure, measuring the concentration value for a value on the abscissa axis of the representation. of y = 0. 5

    The process has a reproducibility of 4.6% (n = 3) calculated as the deviation of the slopes of the calibration lines obtained.
     10
    The detection capacity is 2.3 ± 0.9 µM and the range of measurement concentrations is between 1.9 - 16.6 µM.
     fifteen
    权利要求:
    Claims (7)
    [1]

    1. - Electrode device for the detection of ascorbic acid obtained by screen printing characterized in that it comprises three electrodes (1, 2, 3), each electrode consists of a contact or terminal (1.3, 2.3, 3.3), a section (1.2 , 2.2, 3.2) and an active area (1.1, 2.1, 3.1), being one of reference (1), another working electrode (2), and another auxiliary or counter electrode (3), constituted the terminals and sections with a silver ink, the 10 active areas of the counter electrode and the working electrode with a carbon ink and the active area of the reference electrode with a silver / silver chloride ink.
     fifteen
    [2]
    2. - Electrode device according to claim 1 characterized in that the contacts (1.3, 2.3, 3.3), the sections (1.2, 2.2, 3.2) and the active area of the reference electrode (1,1) are rectangular in shape.
     twenty
    [3]
    3. - Electrode device according to claim 2, characterized in that the active area (1.1) of the reference electrode is rectangular in shape, 2 mm wide and 4 mm long.
     25
    [4]
    4. Electrode device according to claim 1 characterized in that the active area (2.1) of the working electrode (2) is a circle of 4.5 mm in diameter.
     30
    [5]
    5. Electrode device according to claim 1 characterized in that the active area (3.1) of the auxiliary electrode or counter electrode (3) has an arc shape.
     35
    [6]
    6. Process for manufacturing the device according to claim 1, characterized in that it comprises the following steps:
    -Screen printing of a sheet of polyester with 5 silver ink with the shapes of the contacts (1.3, 2.3, 3.3), and the sections (1.2, 2.2, 3.2) of the three electrodes (1, 2, 3).
    -Screen printing of the active surfaces of the working electrode (2.1) and the counter electrode (3. 1) 10 with carbon ink.
    -Screen printing of the active area of the reference electrode (1.1) with Ag / AgCl.
    -Screen printing of an insulator covering the device except the active areas (1.1, 2.1, 3.1) of the electrodes and the contacts (1.2, 2.2, 3.2) of the electrodes.

    [7]
    7. Use of the device according to claim 1 characterized in that it comprises the following steps:
    - connection of the contacts (1.3, 2.3, 3.3) of the electrodes (1, 2, 3) to a potentiostat to apply the appropriate potential to the electrodeodical system 25
    - modification of the working electrode with gold na-noparticles. The deposition of the gold na-noparticles (AuNPs) on the working electrode is carried out electrochemically de-30 placing in the electrode a 100 µL drop of 0.1 mM solution of HAuCl4 prepared in 0.5 M H2SO4, and applying a poten- constant cial of +0.18 V vs Ag / AgCl SPE for 50 seconds. 35
    - introduction of the electrode device in the electrochemical cell.
    - Adding 5 mL of the sample of ascorbic acid in the presence of a 100 mM buffer solution of KH2PO4 and 100 mM of KCl of pH = 7, subjecting the solution to a constant stirring speed.
     10
    - application of a potential of +0.2 V vs Ag / AgClSPE.
    - realization of successive additions of standard solutions of ascorbic acid by measuring the intensities recorded after each addition.
    - representation of the intensities against ascorbic concentrations to build the calibration line. twenty
    - calculation of the concentration of the sample of ascorbic acid problem by measuring on the abscissa axis of the representation the value of the concentration for a value of y = 0 according to the known standard addition method.

     30

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    同族专利:
    公开号 | 公开日
    ES2660516B1|2018-10-26|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    US20070256944A1|2006-04-14|2007-11-08|Tamkang University|Method and electrochemical sensing strip with screen-printed three electrodes for determining concentration of dissolved oxygen in a solution|
    US20110139636A1|2009-12-14|2011-06-16|Lai Rebecca Y|Gold-plated screen-printed electrodes and their use as electrochemical sensors|
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