Slow injection analysis

There are many potential advantages to kinetic methods of analysis, perhaps the most important of which is the ability to use chemical reactions that are slow to reach equilibrium. In this chapter we examine three techniques that rely on measurements made while the analytical system is under kinetic rather than thermodynamic control chemical kinetic techniques, in which the rate of a chemical reaction is measured radiochemical techniques, in which a radioactive element s rate of nuclear decay is measured and flow injection analysis, in which the analyte is injected into a continuously flowing carrier stream, where its mixing and reaction with reagents in the stream are controlled by the kinetic processes of convection and diffusion. 

Since mass spectrometry is a rapid analytical technique, the sample throughput will be -20 per hour when using FAB/LSIMS and >60 per hour when using flow injection analysis with APCI or ESI. The rate-limiting step for FAB/LSIMS is the time (1 to 2 min) required to introduce the direct insertion probe through the vacuum interlock. During LC/MS and LC/MS/MS, the slow step is the time required for chromatographic separation. 

There have been several approaches to overcome the traditionally slow SEC separations, which are caused by the diffusion processes in SEC columns. Most of them are column-related (see High-Speed SEC Columns, Small Particle Technology, and Smaller SEC Column Dimensions ) one utilizes the column void volume (cf. Overlaid Injections ), while another replaces separation with simplified sample preparation (see Flow Injection Analysis ). Cloning existing methods and instrumentation is also reviewed with respect to the potential time gain (see Cloning of SEC Systems ). Benefits and limitations of each method are summarized in Table 1. 

In 1985, mono-segmented flow analysis was proposed [64] as a means of achieving extended sample incubation times without excessive sample dispersion. The sample was inserted between two air bubbles into an unsegmented carrier stream therefore the innovation combined the favourable characteristics of both segmented and unsegmented flow systems. Further development revealed other potential applications, especially with regard to relatively slow chemical reactions, flow titrations, sample introduction to atomic absorption spectrometers, liquid-liquid extraction and multi-site detection (Chapters 7 and 8). This innovation was also referred to as segmental flow injection analysis [65]. 

Flow-Injection analysis is suitable for use with chemical systems with very different reaction rates. Thus, fast reactions are usually treated by normal FIA, whereas slow processes are generally dealt with by other modes such as zone trapping [64] or stopped-flow techniques [61]. 

Recently a decreased level of CE activity has been noticed with a shift of attention towards other separation techniques such as electrochromatography. CE is apparently not more frequently used partly because of early instrumental problems associated with lower sensitivity, sample injection, and lack of precision and reliability compared with HPLC. CE has slumped in many application areas with relatively few accepted routine methods and few manufacturers in the market place. While the slow acceptance of electrokinetic separations in polymer analysis has been attributed to conservatism [905], it is more likely that as yet no unique information has been generated in this area or eventually only the same information has been gathered in a more efficient manner than by conventional means. The applications of CE have recently been reviewed [949,950] metal ion determination by CE was specifically addressed by Pacakova et al. [951]. 

As described for equilibria before, separation techniques like filter binding assays or pull down methods can be applied to separate educts and products of slow reactions too. Here modern HPLC systems with autosampler and time programmed injection offer a convenient approach for the analysis of reactions in the time scale of minutes and hours. 

The primary objective of integration plot analysis is to analyze the data on influx of the test substrate from the circulating blood to the retina (i.e., blood-to-retina direction) across the BRB after intravenous administration of the test substrate. The advantage of this approach is that it allows reliable determination of the retinal uptake (i.e., clearance) of the test substrate which has a slow permeability across the BRB [28], On the other hand, due to the intravenous injection, interference by endogenous substrates and plasma-protein binding of the test substrate can produce an unseemingly low estimate of the retinal uptake. 

Chlorine was injected periodically from a cylinder containing 5% CI2 gas in dry nitrogen. The gas mixture was sparged into the system through two 30 mm fritter glass discs of medium porosity. Chlorine dissipation rates were found to be slow and chlorine levels could be maintained reasonably constant ( 5%) by injecting fresh gas at about 12 hour intervals. Flow rates and injection times were established by analysis of chamber contents. 

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