Detector amperometric

This type of detector is used in ion chromatography for the detection of inorganic anions (e.g., S04, PO43 ), some inorganic cations (e.g Ca +, Mg2+), and some ionised organic acids. This is due to the fact that all ions are electrically conducting. Conductivity detectors are based on the conductance of an eluent prior to and during the elution of the analyte from the column. 

This type of detector measures the current generated in an electrochemical cell at a fixed applied potential by the reduction or oxidation and an eluted analyte at the surface of a microelectrode. Sometimes this is called the counterelectrode (usually made of gold, platinum, or a glassy carbon) and auxiliary or working electrode and a reference electrode, usually Ag/AgCl. The mobile phase used will act as the supporting electrolyte for the redox reactions therefore, its composition is restricted to aqueous solvent mixtures. Amperometric detection is used for ions that have pKa values of greater than 7 and thus cannot be detected by conductivity detectors (because the formed products are weakly dissociated). 

The potential required or the reduction or oxidation of the analyte being detected is applied between the auxiliary and the reference electrode. The microelectrode acts as the counterelectrode and functions to protect the potential and to prevent damage to the reference electrode. When a mixture containing ions (electrochemically active) flows through the measuring cell within the instrument, it is partially reduced or oxidised. This in turn produces a cathodic or anodic current that is proportional to the concentration of the analyte. 

---

The use of an amperometric detector is emphasized in this experiment. Hydrodynamic voltammetry (see Chapter 11) is first performed to identify a potential for the oxidation of 4-aminophenol without an appreciable background current due to the oxidation of the mobile phase. The separation is then carried out using a Cjg column and a mobile phase of 50% v/v pH 5, 20 mM acetate buffer with 0.02 M MgCl2, and 50% v/v methanol. The analysis is easily extended to a mixture of 4-aminophenol, ascorbic acid, and catechol, and to the use of a UV detector. 

HPLC DETERMINATION OF PHENOLS WITH PHOTOMETRIC AND AMPEROMETRIC DETECTORS... 

For selective estimation of phenols pollution of environment such chromatographic methods as gas chromatography with flame-ionization detector (ISO method 8165) and high performance liquid chromatography with UV-detector (EPA method 625) is recommended. For determination of phenol, cresols, chlorophenols in environmental samples application of HPLC with amperometric detector is perspective. Phenols and chlorophenols can be easy oxidized and determined with high sensitivity on carbon-glass electrode. 

The comparison of analytical characteristics HPLC methods of determination of phenols with application amperometric and photometric detectors was caiiy out in this work. Experiment was executed with use liquid chromatograph Zvet-Yauza and 100 mm-3mm 150mm-3mm column with Silasorb C18 (5 10 p.m). With amperometric detector phenols were detected in oxidizing regime on glass-cai bon electrodes. With photometric detector phenols were detected at 254 nm. 

The amperometric detector is currently the most widely used electrochemical detector, having the advantages of high sensitivity and very small internal cell volume. Three electrodes are used ... 

Despite their higher sensitivity and relative cheapness compared with ultraviolet detectors, amperometric detectors have a more limited range of applications, being often used for trace analyses where the ultraviolet detector does not have sufficient sensitivity. 

Depending on their conversion efficiency, electrochemical detectors can be divided into two categories those that electrolyze only a negligible fraction (0.1-5%) of the electroactive species passing through the detector (amperometric detectors), and those for which the conversion efficiency approaches 100% (coulo-metric detectors). Unfortunately, the increased conversion efficiency of the analyte is accompanied by a similar increase for the electrolyte (background) reactions, and no lowering of detection limits is reahzed. 

The most widely used amperometric detectors are based on the thin-layer and wall-jet configurations (Figure 3-22). The thin-layer cell relies on a thin layer of solution that flows parallel to the planar electrode surface, which is imbedded in a... 

FIGURE 3-23 Schematic of a carbon-fiber amperometric detector for capillary electrophoresis A, fused silica capillary B, eluent drop C, stainless steel plate RE, reference electrode WE, working electrode, AE, auxiliary electrode. (Reproduced with permission from reference 58.)... 

Example 3-6 Flow analysis of a urine sample at a thin-layer amperometric detector, with a flow rate of 1.25mLmin yielded a limiting current value of 1.6 pA for its unknown uric acid content. A larger current of 2.4 pA was observed for a sample containing 1 x 10 4 M uric acid and flowing at a rate of 0.9 mL min. Calculate the original concentration of uric acid in the sample. 

Figure 1. HPLC analysis of product progression during hydrolysis of 0.25 % polygalacturonate by PGII. Aliquots were withdrawn from the reaction mixture at timed intervals and reactions were stopped by raising the pH of the sample to pH 8.0 by mixing with 1 volume 25 mM Na-phosphate pH 9.5. Gl to G5 indicate the oligogalacturonates with corresponding degree of polymerization. The vertical axis shows the responce of the pulsed amperometric detector and the horizontal axis the elution time. Times of sampling are indicated above the trace.

Figure 5. Selected HPLC elution profile of products obtained after incubation of 0.25% polygalacturonate with PGII, upper trace, and PGII H223A, lower trace, respectively, demonstrating the effect of the mutation on catalysis. G1 to G3 indicate the peaks of the corresponding oligogalacturonates. IS indicates the internal standard, glucuronate. The vertical axis shows the pulsed amperometric detector response while the horizontal axis shows the retention time.

While the terms amperometric detection and coulometric detection have come into use to describe detectors of less than 100% efficiency and 100% efficiency respectively, these terms are actually misnomers. An amperometric detector is any electrochemical detector where current is plotted as a function of time, regardless of the conversion efficiency. A coulometric detector is any electrochemical detector where charge is plotted as a function of time, again regardless of the conversion efficiency. Preferred terminology should be high efficiency and low efficiency detectors to describe the two situations. 

Enzyme linked electrochemical techniques can be carried out in two basic manners. In the first approach the enzyme is immobilized at the electrode. A second approach is to use a hydrodynamic technique, such as flow injection analysis (FIAEC) or liquid chromatography (LCEC), with the enzyme reaction being either off-line or on-line in a reactor prior to the amperometric detector. Hydrodynamic techniques provide a convenient and efficient method for transporting and mixing the substrate and enzyme, subsequent transport of product to the electrode, and rapid sample turnaround. The kinetics of the enzyme system can also be readily studied using hydrodynamic techniques. Immobilizing the enzyme at the electrode provides a simple system which is amenable to in vivo analysis. 

A composite polymer membrane has also been used as an effective amperometric detector for ion exchange chromatography [42] and showed detection limits similar to those obtained with a conductivity detector. An advantage of the amperometric detector based on micro-ITIES over the conductometric detector is that selectively can be tailored by proper choice of the ionophore. For instance, the selectivity of the membrane toward ammonium in the presence of an excess of sodium could be substantially increased by introducing an ammonium-selective ionophore (such as valinomycin) in the gel membrane [42]. 

Because process mixtures are complex, specialized detectors may substitute for separation efficiency. One specialized detector is the array amperometric detector, which allows selective detection of electrochemically active compounds.23 Electrochemical array detectors are discussed in greater detail in Chapter 5. Many pharmaceutical compounds are chiral, so a detector capable of determining optical purity would be extremely useful in monitoring synthetic reactions. A double-beam circular dichroism detector using a laser as the source was used for the selective detection of chiral cobalt compounds.24 The double-beam, single-source construction reduces the limitations of flicker noise. Chemiluminescence of an ozonized mixture was used as the principle for a sulfur-selective detector used to analyze pesticides, proteins, and blood thiols from rat plasma.25 Chemiluminescence using bis (2,4, 6-trichlorophenyl) oxalate was used for the selective detection of catalytically reduced nitrated polycyclic aromatic hydrocarbons from diesel exhaust.26... 

Tabata, S. and Dohi, Y., An assay for oligo-(l—>4) —5(1—>4)-glucantransferase activity in the glycogen debranching enzyme system by using HPLC with a pulsed amperometric detector, Carb. Res., 230,179, 1992. 

For many applications, diode array detection has become routine. A photodiode array was used for simultaneous detection of 100 capillaries in zone electrophoresis and micellar electrokinetic chromatography (MEKC).1516 Deflection of a laser beam by acoustic waves was reported as a means to scan six capillary channels on a microchip.17 The design of a low-noise amperometric detector for capillary electrophoresis has been reported.18... 

Nonspectroscopic detection schemes are generally based on ionisation (e.g. FID, PID, ECD, MS) or thermal, chemical and (electro)chemical effects (e.g. CL, FPD, ECD, coulometry, colorimetry). Thermal detectors generally exhibit a poor selectivity. Electrochemical detectors are based on the principles of capacitance (dielectric constant detector), resistance (conductivity detector), voltage (potentiometric detector) and current (coulometric, polarographic and amperometric detectors) [35]. 

One type of ec detector (the coulometric detector) reacts all of the electroactive solute passing through it. This type has never become very popular (there is only one on the market at the moment). Another type (the amperometric detector) reacts a much smaller quantity of the solute, less than 1%. The currents observed with these detectors are very small (nanoamps), but such currents are not too difficult to measure and the detector has a high sensitivity, considerably higher than that of uv/visible absorbance detectors although not as good as fluorescence detectors. Noise equivalent concentrations of about 10, 0g cm-3 have been obtained in favourable cases. Another advantage of these detectors is that they can be made with a very small internal volume. 

Fig. 2.4j is a simplified diagram of an amperometric detector. Three electrodes are used, called working, auxiliary and reference electrodes (we ae and re). The we is the electrode at which the electroactivity is monitored, and the re, usually a silver-silver chloride electrode, provides a stable and reproducible voltage to which the potential of the we can be referenced. The ae, usually stainless steel, is a current-carrying electrode. 

Occupational air Passage of filtered air through midget impinger containing NaOH conversion of NaCN to sodium formate. Optional ion exchange clean-up. Ion-chromatography/ amperometric detector (HCN) 5-10 ppm 100-109 at 5-20 ppm Dolzine et al. 1982... 

In short, the amperometric detector is presently considered to be the best electrochemical detector having the following distinct advantages, such as ... 

Figure 3.8 Amperometric detectors (a) measure the current that flows between the working electrode, usually a glassy carbon electrode, and a reference electrode, at a fixed voltage, usually close to the discharge potential for the compound. Coulometric detectors (b) are less common and are designed with a porous carbon flow cell so that all the analyte reacts in the cell, the amount of current consumed during the process being proportional to the amount of the substance.

A comparative study of the analysis of aliphatic amines by GC-FID, GC-TSD and HPLC with refractive index detector (RID), using isopropylamine as internal standard, gave good results in all cases. Determination of trimethylamine oxide by HPLC with a pulsed amperometric detector was problematic136. 

---