Graduate Thesis Or Dissertation
 

Development of automated flow injection apparatus and a novel flow cell for chemi- and bioluminescence determinations

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  • A practical and versatile instrument has been developed both to optimize the chemistry of bio- and chemiluminescence reactions and to measure ultra-trace quantities of associated analytes. The instrument consists of a pneumatic flow injection system linked to a detection cell and a signal processing and readout system. The disk-like cell is defined by a pair of transparent polycarbonate plates and an "0-ring" seal. The lower plate is fitted with a reflective surface. Relative to conventional cells, this design affords a high light collection efficiency due to the large volume element viewed by an end-on photomultiplier detector. Rapid mixing of reagents within the cell is brought about by injection through concentric ports of a commercial burner assembly at a point immediately below the detector. The signal processing and readout system is interfaced to an IBM compatible personal computer and appropriate software was written to automate the instrument and to acquire, store and manipulate luminescence data. With this instrumentation, the chemistry of marine bacteria biolumin-escence was optimized for the determination of cis-11-hexadecenal and, ostensibly, for both the quantification of aldehyde insect pheromones and potential use in the control of insect pests. With the optimized conditions, cis- 11-hexadecenal was determined to 7 fmol. This value is more than an order of magnitude lower than detection limits for aldehyde pheromones reported in the literature. In this research, the less ideal substrates undecanal and heptanal were determined to 570 fmol and 65 pmol, respectively. Marine bacteria bioluminescence was used to quantify several epoxide analytes derivatized to aldehydes. 1,2-epoxyhexadecane and 1,2- epoxytetradecane were determined to 55 and 51 fmol, respectively. 1,2- epoxyoctane and cis-7,8-epoxy-2-methyloctadecane were determined to 100 and 3 pmol, respectively. The latter compound is the sex pheromone of the gypsy moth (Lymantria dispar), a well-known and serious agricultural pest. Epoxides have not been quantified previously with either chemi- or bioluminescence. The instrument was modified for use with corrosive solutions and for possible interfacing with a high performance liquid chromatograph. Lophine chemiluminescence was optimized for the analysis of Cr(VI) samples. With the optimized conditions, aqueous solutions of Cr(VI) were determined to 50 μg /L. A plausible explanation is offered for the dependence of lophine chemiluminescence on the concentration of the chromium species.
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