Integration methods for enzymatic analysis response curves which show maxima, minima or inflections Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/9s161863x

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  • Integration methods for enzymatic reactions were applied to transducer response vs. time curves which exhibit maxima, minima or inflections. The difference between the integral of the time dependent response and the initial response, often zero, is related to the initial enzyme activity or substrate concentration. Fluorescence response curves which exhibit a maximum due to pre- and postfilter effects at elevated concentrations of a time dependent fluorescence indicator species result in integrals which may not be unique to one analyte concentration. It was found that augmenting the integral data with the time of the maximum permitted correlation of each integral with a unique analyte concentration or enzyme activity. Integrals of response curves which exhibit minima also correlate with unique analyte concentrations. At increased concentration of analyte, however, the minimum may broaden into a minimum plateau and the time of the minimum may supplement the integral data as a more sensitive measure of analyte concentration. Integrals of enzymatic response curves with inflection points correlate with unique analyte concentrations. Integration methods were applied to both real and ideal systems. Theoretical rate equations were used to generate families of idealized response curves exhibiting maxima, minima and inflections. Curves were generated at either fixed initial substrate concentration with variable enzyme activity or at fixed initial enzyme activity with variable substrate concentration. Difference integrals were calculated and plotted as a function of initial substrate concentration or initial enzyme activity. Enzymatic analyses were also performed using systems which produce families of response curves exhibiting maxima, minima and inflections. Difference integrals were calculated for these curves and related to initial enzyme activity or substrate concentration.
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