Amperometry

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Amperometry in chemistry is detection of ions in a solution based on electric current or changes in electric current.

Contents

Amperometry is used in electrophysiology to study vesicle release events using a carbon fiber electrode. Unlike patch clamp techniques, the electrode used for amperometry is not inserted into or attached to the cell, but brought in close proximity of the cell. The measurements from the electrode originate from an oxidizing reaction of a vesicle cargo released into the medium. Another technique used to measure vesicle release is capacitive measurements.

History

Electrochemical or amperometric detection as it was first used in ion chromatography was single-potential or DC amperometry, useful for certain electrochemically active ions such as cyanide, sulfite, and iodide. The development of pulsed amperometric detection (PAD) for analytes that fouled electrode surfaces when detected eventually helped create a new category of ion chromatography for the determination of carbohydrates. Another advancement, known as integrated amperometry, has increased the sensitivity for other electrochemically active species, such as amines and many compounds that contain reduced sulfur groups, that are sometimes weakly detected by PAD. [1]

It was established that neurotransmitters could be electrochemically detected by placing a carbon electrode into tissue and recording the current from oxidizing neurotransmitters. [2] One of the first measurements was made using an implanted carbon fiber electrode in the neostriatum of rats. [3] Further work was done in chromaffin cells to investigate catecholamine release from large dense core vesicles. [4] [5]

Detection methods

Single-potential amperometry

Any analyte that can be oxidized or reduced is a candidate for amperometric detection. The simplest form of amperometric detection is single-potential, or direct current (DC), amperometry. A voltage (potential) is applied between two electrodes positioned in the column effluent. The measured current changes as an electroactive analyte is oxidized at the anode or reduced at the cathode. Single-potential amperometry has been used to detect weak acid anions, such as cyanide and sulfide, which are problematic by conductometric methods. Another, possibly more important advantage of amperometry over other detection methods for these and other ions, such as iodide, sulfite, and hydrazine, is specificity. The applied potential can be adjusted to maximize the response for the analyte of interest while minimizing the response for interfering analytes [6]

Pulsed amperometry (pulsed amperometric detection, PAD)

An extension of single-potential amperometry is pulsed amperometry, most commonly used for analytes that tend to foul electrodes. Analytes that foul electrodes reduce the signal with each analysis and necessitate cleaning of the electrode. In pulsed amperometric detection (PAD), a working potential is applied for a short time (usually a few hundred milliseconds), followed by higher or lower potentials that are used for cleaning the electrode. The current is measured only while the working potential is applied, then sequential current measurements are processed by the detector to produce a smooth output. PAD is most often used for detection of carbohydrates after an anion exchange separation, but further development of related techniques show promise for amines, reduced sulfur species, and other electroactive compounds.

Principle

In order to record vesicle fusion, a carbon fiber electrode is brought close to the cell. The electrode is held at a positive potential, and when the cargo from a fused vesicle is near the electrode, oxidation of the cargo transfers electrons to the electrode. This causes a spike, the size of which can be used to estimate the number of vesicles, and the frequency gives information about the release probability. [7]

Related Research Articles

Electrochemistry Branch of chemistry

Electrochemistry is the branch of physical chemistry concerned with the relationship between electrical potential difference, as a measurable and quantitative phenomenon, and identifiable chemical change, with the potential difference as an outcome of a particular chemical change, or vice versa. These reactions involve electrons moving via an electronically-conducting phase between electrodes separated by an ionically conducting and electronically insulating electrolyte.

Electrophysiology Study of the electrical properties of biological cells and tissues.

Electrophysiology is the branch of physiology that studies the electrical properties of biological cells and tissues. It involves measurements of voltage changes or electric current or manipulations on a wide variety of scales from single ion channel proteins to whole organs like the heart. In neuroscience, it includes measurements of the electrical activity of neurons, and, in particular, action potential activity. Recordings of large-scale electric signals from the nervous system, such as electroencephalography, may also be referred to as electrophysiological recordings. They are useful for electrodiagnosis and monitoring.

A biosensor is an analytical device, used for the detection of a chemical substance, that combines a biological component with a physicochemical detector. The sensitive biological element, e.g. tissue, microorganisms, organelles, cell receptors, enzymes, antibodies, nucleic acids, etc., is a biologically derived material or biomimetic component that interacts with, binds with, or recognizes the analyte under study. The biologically sensitive elements can also be created by biological engineering. The transducer or the detector element, which transforms one signal into another one, works in a physicochemical way: optical, piezoelectric, electrochemical, electrochemiluminescence etc., resulting from the interaction of the analyte with the biological element, to easily measure and quantify. The biosensor reader device connects with the associated electronics or signal processors that are primarily responsible for the display of the results in a user-friendly way. This sometimes accounts for the most expensive part of the sensor device, however it is possible to generate a user friendly display that includes transducer and sensitive element. The readers are usually custom-designed and manufactured to suit the different working principles of biosensors.

Cyclic voltammetry

Cyclic voltammetry (CV) is a type of potentiodynamic electrochemical measurement. In a cyclic voltammetry experiment, the working electrode potential is ramped linearly versus time. Unlike in linear sweep voltammetry, after the set potential is reached in a CV experiment, the working electrode's potential is ramped in the opposite direction to return to the initial potential. These cycles of ramps in potential may be repeated as many times as needed. The current at the working electrode is plotted versus the applied voltage to give the cyclic voltammogram trace. Cyclic voltammetry is generally used to study the electrochemical properties of an analyte in solution or of a molecule that is adsorbed onto the electrode.

Amperometric titration refers to a class of titrations in which the equivalence point is determined through measurement of the electric current produced by the titration reaction. It is a form of quantitative analysis.

Voltammetry Set of electroanalytical methods

Voltammetry is a category of electroanalytical methods used in analytical chemistry and various industrial processes. In voltammetry, information about an analyte is obtained by measuring the current as the potential is varied. The analytical data for a voltammetric experiment comes in the form of a voltammogram which plots the current produced by the analyte versus the potential of the working electrode.

Chronoamperometry

Chronoamperometry is an electrochemical technique in which the potential of the working electrode is stepped and the resulting current from faradaic processes occurring at the electrode is monitored as a function of time. The functional relationship between current response and time is measured after applying single or double potential step to the working electrode of the electrochemical system. Limited information about the identity of the electrolyzed species can be obtained from the ratio of the peak oxidation current versus the peak reduction current. However, as with all pulsed techniques, chronoamperometry generates high charging currents, which decay exponentially with time as any RC circuit. The Faradaic current - which is due to electron transfer events and is most often the current component of interest - decays as described in the Cottrell equation. In most electrochemical cells this decay is much slower than the charging decay-cells with no supporting electrolyte are notable exceptions. Most commonly a three electrode system is used. Since the current is integrated over relatively longer time intervals, chronoamperometry gives a better signal to noise ratio in comparison to other amperometric techniques.

Flow battery Type of electrochemical cell

A flow battery, or redox flow battery, is a type of electrochemical cell where chemical energy is provided by two chemical components dissolved in liquids that are pumped through the system on separate sides of a membrane. Ion exchange occurs through the membrane while both liquids circulate in their own respective space. Cell voltage is chemically determined by the Nernst equation and ranges, in practical applications, from 1.0 to 2.43 volts.

Flame ionization detector Type of gas detector used in gas chromatography

A flame ionization detector (FID) is a scientific instrument that measures analytes in a gas stream. It is frequently used as a detector in gas chromatography. The measurement of ion per unit time make this a mass sensitive instrument. Standalone FIDs can also be used in applications such as landfill gas monitoring, fugitive emissions monitoring and internal combustion engine emissions measurement in stationary or portable instruments.

Potentiometric titration is a technique similar to direct titration of a redox reaction. It is a useful means of characterizing an acid. No indicator is used; instead the potential is measured across the analyte, typically an electrolyte solution. To do this, two electrodes are used, an indicator electrode and a reference electrode. Reference electrodes generally used are hydrogen electrodes, calomel electrodes, and silver chloride electrodes. The indicator electrode forms an electrochemical half cell with the interested ions in the test solution. The reference electrode forms the other half cell.

Electroanalytical methods are a class of techniques in analytical chemistry which study an analyte by measuring the potential (volts) and/or current (amperes) in an electrochemical cell containing the analyte. These methods can be broken down into several categories depending on which aspects of the cell are controlled and which are measured. The three main categories are potentiometry, coulometry, and voltammetry.

Electrochemiluminescence Emission of light from electrochemical reactions

Electrochemiluminescence or electrogenerated chemiluminescence (ECL) is a kind of luminescence produced during electrochemical reactions in solutions. In electrogenerated chemiluminescence, electrochemically generated intermediates undergo a highly exergonic reaction to produce an electronically excited state that then emits light upon relaxation to a lower-level state. This wavelength of the emitted photon of light corresponds to the energy gap between these two states. ECL excitation can be caused by energetic electron transfer (redox) reactions of electrogenerated species. Such luminescence excitation is a form of chemiluminescence where one/all reactants are produced electrochemically on the electrodes.

Electrochemical gas sensors are gas detectors that measure the concentration of a target gas by oxidizing or reducing the target gas at an electrode and measuring the resulting current.

Scanning electrochemical microscopy (SECM) is a technique within the broader class of scanning probe microscopy (SPM) that is used to measure the local electrochemical behavior of liquid/solid, liquid/gas and liquid/liquid interfaces. Initial characterization of the technique was credited to University of Texas electrochemist, Allen J. Bard, in 1989. Since then, the theoretical underpinnings have matured to allow widespread use of the technique in chemistry, biology and materials science. Spatially resolved electrochemical signals can be acquired by measuring the current at an ultramicroelectrode (UME) tip as a function of precise tip position over a substrate region of interest. Interpretation of the SECM signal is based on the concept of diffusion-limited current. Two-dimensional raster scan information can be compiled to generate images of surface reactivity and chemical kinetics.

A biotransducer is the recognition-transduction component of a biosensor system. It consists of two intimately coupled parts; a bio-recognition layer and a physicochemical transducer, which acting together converts a biochemical signal to an electronic or optical signal. The bio-recognition layer typically contains an enzyme or another binding protein such as antibody. However, oligonucleotide sequences, sub-cellular fragments such as organelles and receptor carrying fragments, single whole cells, small numbers of cells on synthetic scaffolds, or thin slices of animal or plant tissues, may also comprise the bio-recognition layer. It gives the biosensor selectivity and specificity. The physicochemical transducer is typically in intimate and controlled contact with the recognition layer. As a result of the presence and biochemical action of the analyte, a physico-chemical change is produced within the biorecognition layer that is measured by the physicochemical transducer producing a signal that is proportionate to the concentration of the analyte. The physicochemical transducer may be electrochemical, optical, electronic, gravimetric, pyroelectric or piezoelectric. Based on the type of biotransducer, biosensors can be classified as shown to the right.

Fast-scan cyclic voltammetry

Fast-scan cyclic voltammetry (FSCV) is cyclic voltammetry with a very high scan rate (up to 1×106 V·s−1). Application of high scan rate allows rapid acquisition of a voltammogram within several milliseconds and ensures high temporal resolution of this electroanalytical technique. An acquisition rate of 10 Hz is routinely employed.

Electrochemical stripping analysis

Electrochemical stripping analysis is a set of analytical chemistry methods based on voltammetry or potentiometry that are used for quantitative determination of ions in solution. Stripping voltammetry have been employed for analysis of organic molecules as well as metal ions. Carbon paste, glassy carbon paste, and glassy carbon electrodes when modified are termed as chemically modified electrodes and have been employed for the analysis of organic and inorganic compounds.

Semi-solid flow battery

A semi-solid flow battery is a type of flow battery using solid battery active materials or involving solid species in the energy carrying fluid. A research team in MIT proposed this concept using lithium-ion battery materials. In such a system, both positive (cathode) and negative electrode (anode) consist of active material particles with carbon black suspended in liquid electrolyte. Active material suspensions are stored in two energy storage tanks. The suspensions are pumped into the electrochemical reaction cell when charging and discharging. This design takes advantage of both the designing flexibility of flow batteries and the high energy density active materials of lithium-ion batteries.

Single-Entity Electrochemistry (SEE) refers to the electroanalysis of an individual unit of interest. A unique feature of SEE is that it unifies multiple different branches of electrochemistry. Single-Entity Electrochemistry pushes the bounds of the field as it can measure entities on a scale of 100 microns to angstroms. Single-Entity Electrochemistry is important because it gives the ability to view how a single molecule, or cell, or "thing" affects the bulk response, and thus the chemistry that might have gone unknown otherwise. The ability to monitor the movement of one electron or ion from one unit to another is valuable, as many vital reactions and mechanisms undergo this process. Electrochemistry is well suited for this measurement due to its incredible sensitivity. Single-Entity Electrochemistry can be used to investigate nanoparticles, wires, vesicles, nanobubbles, nanotubes, cells, and viruses, and other small molecules and ions. Single-entity electrochemistry has been successfully used to determine the size distribution of particles as well as the number of particles present inside a vesicle or other similar structures

Screen-printed electrodes (SPEs) are electrochemical measurement devices that are manufactured by printing different types of ink on plastic or ceramic substrates, allowing quick in-situ analysis with high reproducibility, sensitivity and accuracy. The composition of the different inks used in the manufacture of the electrode determines its selectivity and sensitivity. This fact allows the analyst to design the most optimal device according to its purpose.

References

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  2. Kissinger PT, Hart JB, Adams RN (May 1973). "Voltammetry in brain tissue--a new neurophysiological measurement". Brain Research. 55 (1): 209–13. doi:10.1016/0006-8993(73)90503-9. PMID   4145914.
  3. Gonon F, Cespuglio R, Ponchon JL, et al. (April 1978). "In vivo continuous electrochemical determination of dopamine release in rat neostriatum". Comptes Rendus de l'Académie des Sciences, Série D (in French). 286 (16): 1203–6. PMID   96981.
  4. Leszczyszyn DJ, Jankowski JA, Viveros OH, Diliberto EJ, Near JA, Wightman RM (September 1990). "Nicotinic receptor-mediated catecholamine secretion from individual chromaffin cells. Chemical evidence for exocytosis". The Journal of Biological Chemistry. 265 (25): 14736–7. doi: 10.1016/S0021-9258(18)77173-1 . PMID   2394692.
  5. Wightman RM, Jankowski JA, Kennedy RT, et al. (December 1991). "Temporally resolved catecholamine spikes correspond to single vesicle release from individual chromaffin cells". Proceedings of the National Academy of Sciences of the United States of America. 88 (23): 10754–8. Bibcode:1991PNAS...8810754W. doi: 10.1073/pnas.88.23.10754 . PMC   53009 . PMID   1961743.
  6. Settle, F. (Ed.). (1997). Handbook of Instrumental Techniques for Analytical Chemistry (1 ed.). Prentice Hall.
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