Neurotransmitters, electrochemical detection

Biogenic amines are of great interest to researchers because of their potential roles in several psychiatric and neurological disorders. They include dopamine (DA), noradrenaline (NA), 5-hydroxytryptamine (5-HT, serotonin), histamine, and trace amines such as 2-phenylethylamine (PEA), tyramine, octopamine, phenylethanolamine, and tryptamine (Coutts and Baker, 1982). Although GC assays for DA, NA, and 5-HT are available, HPLC analysis with electrochemical detection has for many years now been the method of choice for analysis of these neurotransmitter amines. 

Carbon nanotube modified electrodes The electrochemical detection of the neurotransmitter dopamine is complicated by the high concentration of biologically coexisting ascorbic acid, which has an oxidation potential lying very close to dopamine s at solid... 

Research into chemically modified electrodes has led to a number of new ways to build chemical selectivity into films that can be coated onto electrode surfaces. Perhaps the simplest example is the use of the polymer Nafion (see Table 13.2) to make selective electrodes for basic research in neurophysiology [88]. Starting with the pioneering investigations by Ralph Adams, electrochemists have become interested in the electrochemical detection of a class of amine-based neurotransmitters in living organisms. The quintessential example of this class of neurotransmitters is the molecule dopamine, which can be electrochemically oxidized via the following redox reaction ... 

Catecholamines, nerve transmitters monitored in brain and heart patients, are separated on C18 using octane sulfonate ion pairing in 6% An/water (pH 3) with added EDTA and phosphate. Detection can be at UV, 270 nm, or by electrochemical detection at +0.72 V for maximum sensitivity. Other tyrosine and tryptophan metabolite neurotransmitters such as serotonin, VMA, and HMA can be analyzed with ion pairing and EC detection. 

Synaptic neurotransmission in brain occurs mostly by exocytic release of vesicles filled with chemical substances (neurotransmitters) at presynaptic terminals. Thus, neurotransmitter release can be detected and studied by measuring efflux of neurotransmitters from synapses by biochemical methods. Various methods have been successfully employed to achieve that, including direct measurements of glutamate release by high-performance liquid chromatography of fluorescent derivatives or by enzyme-based continuous fluorescence assay, measurements of radioactive efflux from nerve terminals preloaded with radioactive neurotransmitters, or detection of neuropeptides by RIA or ELISA. Biochemical detection, however, lacks the sensitivity and temporal resolution afforded by electrophysiological and electrochemical approaches. As a result, it is not possible to measure individual synaptic events and apply quantal analysis to verify the vesicular nature of neurotransmitter release. 

The suprahypothalamic neurotransmitter level can be assessed by a determination of catecholamines in circumscribed brain areas, the technique requires preparation of frozen tissue and isolation of specific nuclei by the micropunch technique. The catecholamines and indolamines can be measured by a radio-enzymatic methods and by a high-pressure liquid chromatography (HPLC) with electrochemical detection. These mechanistic investigations are mostly initiated due to questions arising from the receptor interaction profile of the drug candidate, they may be required to prove that such receptor interactions truly change the functional state of neurotransmitters (functional expression). Mostly, however, the peripheral effects of such neurotransmitter mechanisms (for instance prolactin secretion) are sufficiently distinct. 

Neurotransmitters have also been detected on microchips using electrochemical detection, eliminating the need for on-chip reactions or derivitization. Amperometric detection, a current change when an analyte passes the detection electrodes, was demonstrated on a microchip for the determination of dopamine concentrations in standard solutions [10], The microdevice developed in this... 

Electrochemical detection is one of the most common methods for neurotransmitter monitoring. Many neurotransmitters are... 

Electrochemical detection involves the induction of a change in redox state (electrolysis) by application of an electrical potential to an electrode (71). Compounds that can be readily detected by this means are termed electroactive. Under physiological conditions, these compounds tend to be in their reduced state in the nervous system because of the rich level of antioxidants (e.g., ascorbic acid) and, thus, can be oxidized by application of a positive potential to the electrode. The evolved electrons are detected at the electrode in the form of electrical current. This current is proportional to the number of electroactive molecules at the surface of the electrode, and therefore it is proportional to their concentration in the bulk solution. By implanting an electrode in the extracellular space close to the release site and detecting changes in the local (extracellular) concentration of the neurotransmitter, neurotransmitter release can be monitored. The key advantage of this approach is the high temporal resolution that can be in the millisecond domain. Neurotransmitters that can be detected this way include dopamine, norepinephrine, epinephrine, serotonin, and melatonin. 

S.6.4.2 Reversed-phase ion-pair liquid chromatography. Reversed-phase ion-pair chromatography is an alternative approach for controlling the retention of ionic compounds. This approach is particularly useful for the separation of amines on silica-based columns and it has had a profound effect on the analysis of this class of compounds. In particular, the combination of reversed-phase ion-pair liquid chromatography and electrochemical detection revolutionized the analysis of neurotransmitters in the brain (Tomlinson et al., 1978 and refs, therein). 

Electrochemical Detection Of Neurotransmitters At Structurally Small Electrodes... 

In this chapter, we aim to review the techniques used in developing structurally small electrodes of different geometries, which were then applied to the detection of neurotransmitters. We will also pay special emphasis on the strategies used to minimize electrode fouling during electrochemical detection of neurotransmitters at these electrodes. A comparison of these methods and possible future directions in the development of structurally small electrodes for detection of neurotransmitters will also be presented. 

Electrochemical Detection o Neurotransmitters a Structurally Small Electrodes 319... 

Electrochemical detection is less widely used than UV absorbance or LIE detection, but has been used successfully by Ewing and coworkers to detect neurotransmitters within subcellular structures. " Electrochemical detection is attractive because it provides a lower LOD than UV absorbance, does not require sample derivatization, and in the case of amperometric detection, can be tuned to specific classes of compounds. Furthermore, unlike UV absorbance in which the sensitivity is dependent on the sample volume (i.e., the pathlength), the LOD of electrochemical detection improves when applied to miniaturized CE systems, since in this case sensitivity is related to contact between the analyte and the electrode surface. In fact, as described above, LODs in the zepto-mole range for the subcellular quantification of dopamine have been reported, with the sensitivity of these results making electrochemical detection the rival of many LIE detectors. However, the... 

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