Electrochemical detector system

D. Perrett, Comparative studies on different electrochemical detector systems in Drug Determination in Therapeutic and Forensic Contexts, Methodological Surveys in Biochemistry, E. Reid and I.D. Wilson, (eds). Plenum, New York, 1984, 14, 111-121. 

Aminosalicylic acid in assessment of reactive oxygen species formation by in vitro Fenton and ozonation reactions and by in vivo ozone-exposure experiments in rats revealed oxidation products as follows salicylic acid, by deamination 2,3-dihydro-xybenzoic acid and 2,5-dihydroxybenzoic acid, from radical or enzymatic hydroxylation 5-amino-2 - hydroxy-N,W-bis(3 - carboxy- 4- hydroxyphenyl)-1,4-benzoquinonediimine, a condensation product of oxidised 5-aminosalicylic acid and 5-amino-2,3,4,6-tetrahydroxybenzoic acid, attributed to hydroxyl radical attack without deamination, identified by high-pressure liquid chromatography electrochemical detector system analysis and by gas chromatography-mass spectrometry analysis of trimethyl silyl derivatives (Kumarathasan et al. 2001). 

The major application of porous, three-dimensional electrodes has been in metal removal from dilute process liquors (Chapter 7) although inorganic and organic dectrosynthesis applications continue to be explored (see, for example, the Dow-Huron cell for production in Chapter S). Many battery and l el cell electrodes utilize an active material which is (or is supported by) a porous, three-dimensional matrix (Chapter 11), while miniature porous electrodes have found use in electrochemical detector systems for high-pressure liquid chromatography analysis (Chapter 12). 

The most common detectors in HPLC are ultraviolet, fluorescence, electrochemical detector and diffractometer. However, despite all improvements of these techniques it seems necessary to have a more selectivity and sensitivity detector for the purposes of the medical analysis. It should be therefore improvements to couple analytical techniques like infrared IR, MS, nuclear magnetic resonance (NMR), inductively coupled plasma-MS (ICP-MS) or biospecific detectors to the LC-system and many efforts have been made in this field. 

Describe clearly the challenges of interfacing electrochemical detectors to capillary electrophoresis separation systems. How can these challenges be overcome ... 

Instead of immobilizing the antibody onto the transducer, it is possible to use a bare (amperometric or potentiometric) electrode for probing enzyme immunoassay reactions (42). In this case, the content of the immunoassay reaction vessel is injected to an appropriate flow system containing an electrochemical detector, or the electrode can be inserted into the reaction vessel. Remarkably low (femtomolar) detection limits have been reported in connection with the use of the alkaline phosphatase label (43,44). This enzyme catalyzes the hydrolysis of phosphate esters to liberate easily oxidizable phenolic products. 

HPLC has been recommended as a cleanup and fiactionation procedure for food samples prior to analysis by GC/ECD (Gillespie and Walters 1986). The advantages over the AOAC-recommended Florisil colunrn are that it is faster, requires less solvent, and gives better resolution. HPLC coupled with various detectors MS, MS/MS, UV/electrochemical detector, or UV/polarographic detection has been tested as a rapid, simplified separation and detection system to replace GC (Betowski and Jones 1988 Clark et al. 1985 Koen and Huber 1970). Recoveries, detection limits, and precisions were generally good, but further work is needed before the techniques are adopted for general use. 

HPAEC analyses were carried out to determine the oligomeric products released from various pectic substrates after depolymerization by the PL isoenzymes. Action pattern analyses for the concerted action of PL isoenzymes utilized 68% esterified pectin as substrate. One-ml reaction mixtures in a buffer system as detailed in section 2.2. comprising 0.5% (w/v) substrate and 5 U of enzyme were incubated for 30 s to 18 h, and then thermoinactivated. Samples of 750 pi were applied to a Carbopac PA-1 (Dionex) column before the carbohydrates were eluted over a period of 70 min using a gradient of 0.2 M KOH, 0.05 M K-acetate to 0.2 M KOH, 0.7 M K-acetate. Detection employed a Pulsed Electrochemical Detector (PED, Dionex) in the integrated amperometry mode according to the manufacturer s recommendations. 

High-performance liquid chromatography (HPLC) with a micellar mobile phase or with a selective pre-column or reaction detection system has also been used to determine alkylenebis(dithiocarbamaes). ° Zineb and mancozeb residues in feed were determined by ion-pair HPLC with ultraviolet (UV) detection at 272 nm. These compounds were converted to water-soluble sodium salts with ethylenediaminetetra-acetic acid (EDTA) and sodium hydroxide. The extracts were ion-pair methylated with tetrabuthylammonium hydrogensulfate (ion-pair reagent) in a chloroform-hexane solvent mixture at pH 6.5-8.S. The use of an electrochemical detector has also been reported. ... 

Brunt, K., Electrochemical detectors for high-performance liquid chromatography and flow analysis systems, Trace Analysis, Vol. 1, Lawrence, J. F., Ed., Academic Press, New York, 1981, 47-120. 

Bergstrom et al. [63] used HPLC for determination of penicillamine in body fluids. Proteins were precipitated from plasma and hemolyzed blood with trichloroacetic acid and metaphosphoric acid, respectively, and, after centrifugation, the supernatant solution was injected into the HPLC system via a 20-pL loop valve. Urine samples were directly injected after dilution with 0.4 M citric acid. Two columns (5 cm x 0.41 cm and 30 cm x 0.41 cm) packed with Zipax SCX (30 pm) were used as the guard and analytical columns, respectively. The mobile phase (2.5 mL/min) was deoxygenated 0.03 M citric acid-0.01 M Na2HP04 buffer, and use was made of an electrochemical detector equipped with a three-electrode thin-layer cell. The method was selective and sensitive for mercapto-compounds. Recoveries of penicillamine averaged 101% from plasma and 107% from urine, with coefficients of variation equal to 3.68 and 4.25%, respectively. The limits of detection for penicillamine were 0.5 pm and 3 pm in plasma and in urine, respectively. This method is selective and sensitive for sulfhydryl compounds. 

Aniline, methyl aniline, 1-naphthylamine, and diphenylamine at trace levels were determined using this technique and electrochemical detection. Two electrochemical detectors (a thin-layer, dual glassy-carbon electrode cell and a dual porous electrode system) were compared. The electrochemical behavior of the compounds was investigated using hydrodynamic and cyclic voltammetry. Detection limits of 15 and 1.5nmol/l were achieved using colourimetric and amperometric cells, respectively, when using an in-line preconcentration step. 

The Hewlett-Packard HP 1049A Electrochemical Detector cell has a unique system for working electrode replacement the working electrode itself (an 8 mm diameter disc) can be removed from its holder, allowing for economic working electrode replacement in case the working electrode surface cannot be restored. 

Due to close proximity of the electrodes in the Hewlett-Packard HP 1049A Electrochemical Detector cell, little conductance is already sufficient for proper functioning of the electrode system. 

The carrier stream is merged with a reagent stream to obtain a chemical reaction between the sample and the reagent. The total stream then flows through a detector (Fig. 1.1 (b)). Although spectrophotometry is the commonly used detector system in this application, other types of detectors have been used, namely fluorometric, atomic absorption emission spectroscopy and electrochemical, e.g. ion selective electrodes. 

Model RR/066 351 and 352 pumps models 750/16 variable-wavelength UV monitor detector 750/11 variable filter UV detector, MPD 880S multiwave plasma detector, 750/14 mass detector 750/350/06 electrochemical detector refractive index detector HPLC columns column heaters, autosamplers, pre-columns derivatization systems, solvent degassers, preparative HPLC systems... 

Trojanowicz, M., Electrochemical Detectors in Automated Analytical Systems , in Modem Techniques in Electroanalysis, Vanysek P. (Ed.), Wiley-Interscience, New York, 1996, pp. 187-239. This chapter contains a fairly thorough discussion of the possible arrangements of electrodes within flow systems, and for a variety of applications. 

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