EVALUATION OF NEW INSTRUMENTS AND KINETIC METHODS FOR DETERMINATIONS OF ANALYTES OF CLINICAL INTEREST
Abstract
This thesis consists of two parts. Part I describes a kinetic method for the determination of acetoacetate in serum and urine. The determination involves the reaction of acetoacetate with glycine and nitroprusside and the monitoring of the pseudo zero-order absorbance versus time curve for 60 seconds and/or the pseudo first-order absorbance versus time curve for longer periods. Results for standard additions to serum and urine are presented for two sets of reaction conditions. The effects of acetone and creatinine as interferences are also described. Part I also incorporates a statistical study of a proposed reaction sequence model by a nonlinear regression technique. Rate constants and a molar absorptivity for the unstable product are estimated and curves generated using these coefficients are compared with experimental data. Part II concerns the evaluation of two instruments, an automated sampler/stopped-flow mixing spectrophotometer and a vidicon-based spectrometer for use in the determination of the substrates cholesterol and glucose and the activity of the enzyme lactate dehydrogenase. The unique sampling and data processing abilities of the automated stopped-flow spectrophotometer are described for enzymatic equilibrium and kinetic cholesterol determinations. The vidicon-based spectrometer is demonstrated to be practical for kinetic determinations of glucose and lactate dehydrogenase. The unique multiwavelength capabilities of the vidicon spectrometer are also demonstrated by application to dual wavelength glucose determinations.
Degree
Ph.D.
Subject Area
Analytical chemistry
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