Water sample assay development, example

Biosensors based on optical fibers as transduction element have been recently reviewed by Wolfbeis [97]. Optical biosensors based on miniattaized SPR or on evanescent field monitoring are not as often found in miniaturized biosensors, especially in comparison to miniaturized electrochemical transducers, yet. Two examples will be given here a miniaturized SPR biosensor by Cullen and co-workers [98] and an evanescent based microchip biosensor by Borchers and co-workers [99]. The best-known SPR biosensor is the BIAcore device from Pharmacia Company, Sweden. It has been on the market for over a decade and is routinely used for hybridization kinetic analyses, specificity analyses, etc. Cullen and co-workers have incorporated a commercially available miniaturized SPR transducer into a field analyzer and developed a competition and inhibition assay for an estrogenic compound in water samples that function as endocrine disrupting compounds (EDCs). 

The sample workup necessary for pesticide residue analysis will vary with each combination of analyte and antibody, each of which may have a different tolerance for the matrix and other factors. The effects of these factors must be considered as with the development of any other analytical technique. Matrix effects for one ELISA system are summarized in Figure 4. While the effect of the matrix on the antibodies in Figure 4 is different for each antibody-solvent-matrix combination, the competitive ELISA standard curves for most of these combinations are similar when expressed as percent of the appropriate control. Some systems may not require extensive adjustment, but this must be tested with each individual system. For example, our molinate assay performs equally well in a variety of water types at high concentrations of molinate (Figure 5). The small difference seen between the buffer and water standard curves in Figure 5 was eliminated by the addition of small amounts of concentrated buffer to water samples to equalize them to the buffer composition. 

Immunoassays offer much potential for rapid screening and quantitative analysis of pesticides in food and environmental samples. However, despite this potential, the field is still dominated by conventional analytical approaches based upon chromatographic and spectrometric methods. We examine some technical barriers to more widespread adoption and utilization of immunoassays, including method development time, amount of information delivered and inexplicable sources of error. Examples are provided for paraquat in relation to exposure assessment in farmworkers and food residue analyses molinate in relation to low-level detection in surface waters and bentazon in relation to specificity and sensitivity requirements built in to the immunizing antigen. A comparison of enzyme-linked immunosorbent assay (ELISA) results with those obtained from conventional methods will illustrate technical implementation barriers and suggest ways to overcome them. 

Different types of organisms such as daphnia, mussels, algae, and fish have been extensively incorporated in toxicity tests for water assessment systems [65], Most of these assays are developed as test systems with few as laboratory-based sensor systems. Membranes with their active enzyme system have also been implemented in the development of toxicity kits and sensors. An example is the MitoScan Kit (Harvard BioScience, Inc., Holliston, MA), which uses fragmented inner mitochondrial membrane vesicles isolated from beef heart (EPA, 2005 [9]). The submito-chondrial particles contain complexes of enzymes responsible for electron transport and oxidative phosphorylation. When specific toxins are in the sample, the enzyme reactions are slowed or inhibited, and these are monitored spectophotometrically at 340 mn. This is still in a bioassay test kit format but may be developed to optical sensor system. 

The need for maximum sample throughput and minimal human interaction within analytical procedures has provided considerable impetus to the development of integrated systems. SPE-LC in-tube SPME followed by ultraviolet (UV) or MS detection and membrane introduction mass spectrometry (MIMS) have both been used to this end. Submersible MIMS systems capable of extended underwater deployment down to 200 m and with a mass range of up to 200 amu have recently come onto the market. Elow injection coupled with MIMS allows fast, near-real-time determination of, for example, phenols in water. Derivatization of the phenols with acetic anhydride can be used to enhance both the selectivity and sensitivity of this method. Other online derivatization procedures are under development with a view to increasing the scope for rapid determination of highly polar compounds that have previously proved difficult to analyze. Large volume injection techniques and developments in enzyme-linked immunosorbent assay (ELISA) technologies... 

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