Electrochemical detection traditional methods

Despite being relatively new technology, aptamers have a tremendous potential and can be envisioned to rival antibodies and other traditional recognition elements for molecular detection and recognition, due to their inherent affinity, selectivity, and ease of synthesis. In addition, the combination of aptasensors with electrochemical detection methods has the added advantage of further cost reduction and miniaturization of such systems. 

Electrochemical DNA biosensors are based on the use of nucleic acids or analogues as biorecognition element and electrochemical techniques for the transduction of the physical chemical signal. Two aspects are essential in the development of hybridization biosensors, sensitivity and selectivity. Traditional methods for detecting the hybridization event are too slow and require special preparation. Therefore, there is an enormous interest in developing new hybridization biosensors, and the electrochemical represent a very good alternative [108]. 

Nanosensors for electrochemical detection have been made for years using more traditional fabrication methods, e.g., pulled platinum strings and carbon fibers. Carbon fibers can be purchased with diameters in the low /am range. These can subsequently be etched in an Ar beam until conically shaped tips are produced with tip diameters between 100 and 500 nm [61]. Similarly, a platinum wire can be heated and pulled in order to create tips of similar diameters. Thick film electrodes made by screen printing [62] have also been shown to find application as transducer in microchaimel systems [63]. 

Electrochemical detection has matured considerably in recent years and is routinely used by many laboratories, often for a very specific biomedical application. The most popular applications include acetylcholine, serotonin, catecholamines, thiols and disulfides, phenols, aromatic amines, macrocycUc antibiotics, ascorbic acid, nitro compounds, hydroxylamines, and carbohydrates. As the last century concluded, it is fair to say that many applications for which LC-EC would be an obvious choice are now pursued with LC-MS-MS. This only became practical in the 1990s and is clearly a more general method applicable to a wider variety of substances. In a similar fashion, LC-MS-MS has also largely supplanted LC-F for new bioanalytical methods. Nevertheless, there remain a number of key applications for these more traditional detectors known for their selectivity (and therefore excellent detection limits). 

For detection of carbohydrates in principle, ultraviolet (UV), laser-induced fluorescence, refractive index, electrochemical, amperometric, and mass spec-trometric detection can be used. Mass spectrometry, with its various ionization methods, has traditionally been one of the key techniques for the structural determination of proteins and carbohydrates. Fast-atom bombardment (FAB) and electrospray ionization (ESI) are the two on-line ionization methods used for carbohydrate analysis. The ESI principle has truly revolutionized the modern mass spectrometry of biological molecules, due to its high sensitivity and ability to record large-molecule entities within a relatively smaU-mass scale. 

Hyphenated instruments consist of an inlet that separates and preconcentrates analytes prior to their introduction into an integral detector. A hyphen can be used to separate the two components when the name is written in abbreviated form—for example, GC-MS. The separation module is the GC and the detection module is the MS. As hyphenated instruments became more common, the hyphen often was dropped, and that is the convention used in this book (i.e., GCMS rather than GC-MS). Separations alone, such as in thin layer chromatography, are not instrumental methods in the traditional sense, but mating a separation technique to a detector creates an instrument that is capable of what is called a hyphenated technique. The basis of most separation modules is selective partitioning, a topic discussed in the previous chapter. However, there is a group of hyphenated instruments in which separation is achieved with electrochemical methods, an increasingly important topic in forensic chemistry. 

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