Pulsed coulometric detection

Figure 27.16B shows the method known as pulsed coulometric detection (PCD). In this case, the current is integrated over a longer period and the time period is an integral number of 16.7-ms segments with typical total integration times of greater than 200 ms. The use of this type of waveform eliminates the most common electrical interference (60 Hz sinusoidal) encountered in pulsed electrochemical detection, and thereby increases the detection limits for most compounds. 

Iodide in urine [39] and catecholamines [40,41] are example of analytes recently detected electrochemically and studied under IPC conditions. Pulsed amperometric detection on a gold electrode was used to detect etimicin [42] and gentamicin [43] in commercial samples, thus avoiding tedious pre-column derivatization. Heterocyclic aromatic amines in soup cubes [44] were determined by a coulometric electrode array detector, and the coulometric detection of a quinone-bearing drug candidate [45] allowed the study of electrochemical properties. 

An ECD measures the current generated by electroactive analytes in the HPLC eluent between electrodes in the flow cell. It offers sensitive detection (pg levels) of catecholamines, neurotransmitters, sugars, glycoproteins, and compounds containing phenolic, hydroxyl, amino, diazo, or nitro functional groups. The detector can be the amperometric, pulsed-amperometric, or coulometric type, with the electrodes made from vitreous or glassy carbon, silver, gold, or platinum, operated in the oxidative or reductive mode. Manufacturers include BSA, ESA, and Shimadzu. 

All of the fat-soluble vitamins, including provitamin carotenoids, exhibit some form of electrochemical activity. Both amperometry and coulometry have been applied to electrochemical detection. In amperometric detectors, only a small proportion (usually <20%) of the electroactive solute is reduced or oxidized at the surface of a glassy carbon or similar nonporous electrode in coulometric detectors, the solute is completely reduced or oxidized within the pores of a graphite electrode. The operation of an electrochemical detector requires a semiaqueous or alcoholic mobile phase to support the electrolyte needed to conduct a current. This restricts its use to reverse-phase HPLC (but not NARP) unless the electrolyte is added postcolumn. Electrochemical detection is incompatible with NARP chromatography, because the mobile phase is insufficiently polar to dissolve the electrolyte. A stringent requirement for electrochemical detection is that the solvent delivery system be virtually pulse-free. 

Although tocopherols and tocotrienols can be detected by UV absorbance at 280 nm, fluorescence detection (excitation 294 nm and emission 326 nm), as shown in Figure 11.3, has proven to be a much more sensitive method. Electrochemical detection such as pulsed amperometric and coulometric (Uspitasari-Nienaber, 2002) has also proven to be sensitive and potentially valuable for the quantitative analysis of tocopherols and Tocotrienols (Abidi, 2000), especially for tocol analysis in blood and serum samples. HPLC mass detectors such as flame-ionization detectors, evaporative light-scattering detectors, and charged aerosol detectors have proven to be valuable for the quantitative analysis of many types of lipids, but because tocols have... 

Several coulometric and pulse techniques are used in electroanalytical chemistry. Rather low detection limits can be achieved, and kinetic and transport parameters can be deduced with the help of these fast and reliable techniques. Since nowadays the pulse sequences are controlled and the data are collected and analyzed using computers, different pulse programs can easily be realized. Details of a wide variety of coulometric and pulse techniques, instrumentation and applications can be found in the following literature controlled current coulometry [6], techniques, apparatus and analytical applications of controlled potential coulometry [7], coulostatic pulse techniques [8], normal pulse voltammetry [9], differential pulse voltammetry [9], and square-wave voltammetry [10]. 

Any of the methods of detection used in liquid chromatography can be used in IC, though some are more useful than others. If the eluent does not affect the detector the need for a suppressor disappears. Common means of detection in IC are ultraviolet (UV) absorption, including indirect absorption electrochemical, especially amperometric and pulsed amperometric and postcolumn derivatization. Detectors atomic absorption spectrometry, chemiluminescence, fluorescence, atomic spectroscopic, refractive index, electrochemical (besides conductivity) including amperometric, coulometric, potentiometric, polaro-graphic, pulsed amperometric, inductively coupled plasma emission spectrometry, ion-selective electrode, inductively coupled plasma mass spectrometry, bulk acoustic wave sensor, and evaporative light-scattering detection. 

Bakker et al. reported this method improving the limit of detection during a thin-layer coulometric analysis [12-14]. An ionophore was doped enhancing the ion-selective ability of a thin-layer membrane. Using a thin-layer coulometry coupling with double-pulse technique, the analysis of nitrate (NOsOi potassium (K" ), Calcium (Ca" ) were successfully undertaken in the scale of micromolar (pM). Furthermore, the advanced compensation of non-faradic charging... 

The amperometric detection uses less than 10% of the analyte in the flow cell, unlike the coulometric detector, and can be operated in a pulsed mode (cydic voltammetry, with a gold working electrode) in addition to the constant potential mode. The pulsed mode helps cleaning the working electrode. 

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