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Since the first report on the ferrocene mediated oxidation of glucose by GOx [69], extensive solution-phase studies have been undertaken in an attempt to elucidate the factors controlling the mediator-enzyme interaction. Although the use of solution-phase mediators is not compatible with a membraneless biocatalytic fuel cell, such studies can help elucidate the relationship between enzyme structure, mediator size, structure and mobility, and mediation thermodynamics and kinetics. For example, comprehensive studies on ferrocene and its derivatives [70] and polypy-ridyl complexes of ruthenium and osmium [71, 72] as mediators of GOx have been undertaken. Ferrocenes have come to the fore as mediators to GOx, surpassing many others, because of factors such as their mediation efficiency, stability in the reduced form, pH independent redox potentials, ease of synthesis, and substitutional versatility. Ferrocenes are also of sufficiently small size to diffuse easily to the active site of GOx. However, solution phase mediation can only be used if the future biocatalytic fuel cell... 

Pure fluids. Carbon dioxide is often the mobile phase of choice for SFC, since it has relatively mild critical parameters, is nontoxic and inexpensive, chemically inert, and is compatible with a wide variety of detectors including the flame ionization detector (FID) used widely in GC and the UV absorbance detector employed frequently in HPLC (7). The usefulness of carbon dioxide as a mobile phase in many instances is somewhat limited, however, because of its nonpolarity (8), and many polar compounds appear to be insoluble in it. For a sample containing polar compounds, pure carbon dioxide may not be the proper mobile phase. The elution of polar compounds is often difficult and the peak shapes for these polar compounds are sometimes poor. This latter difficulty is commonly observed with nonpolar supercritical fluids and may be due to active sites on the stationary phase rather than any inherent deficiency in the fluid itself. 

They should have a polar moiety or a set of polar functional groups that is responsible for the ion recognition process. The rest of the ionophore molecules should contain hydrophobic regions that are compatible with the surrounding membrane matrix. The historical argument that an ionophore molecule must also exhibit a certain mobility within the membrane has been largely disproved by the comparable analytical performance of a number of membrane materials where the ionophore is covalently anchored onto the polymeric backbone [35]. It seems beneficial, however, to at least either have mobile ionophores or mobile ionic sites to guarantee an acceptably low membrane resistance. 

ESR has been useful in studying the influence of dissolved gases on polymer mobility [801]. Stable nitroxyl radicals, such as 2,2,6,6-tetramethylpipe-ridin-l-oxyl (TEMPO) are widely employed as spectroscopic probes for observing binding sites and molecular motion of macromolecules [802]. ESR spectra of the TEMPO free radical in PC film at various temperature and in solution were reported [795]. The TEMPO spin probe method was also used to study diisooctylphthalate (DIOP) plasticiser diffusion in suspension polymerised PVC particles [803]. Similarly, the compatibility Umit of PVAc and di-nonylphthalate (DNP) was studied by means of 2,2-di-n-nonyl-5,5-dimethyl-3-oxazolidinyloxy spin probe ESR measurements and DSC [804] DNP is an effective plasticiser for PVAc for concentrations not exceeding 17 wt.%. According to ESR evidence BBP in PVC forms radicals more easily than DOP [805]. 

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