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Optimization of Nitric Oxide Chemiluminescence Operating Conditions for Measurement of Plasma Nitrite and Nitrate

¹ Vascular Biology Program, Lawson Health Research Institute, London Health Sciences Centre, London, Ontario, N6B 1B8 Canada
Departments of
² Medical Biophysics and
³ Medicine, University of Western Ontario, London, Ontario, N6A 5C1 Canada

^aaddress correspondence to this author at: Department of Medicine, University of Western Ontario, University Hospital 339 Windermere Rd., London, Ontario, N6A 5A5 Canada; fax 519-663-3789, e-mail dfreeman@uwo.ca

The oxidized nitric oxide (NO) metabolites nitrite (NO₂^-) and nitrate (NO₃^-) have been used as markers of NO synthase activity in several clinical and experimental studies including those investigating sepsis (1)(2)(3)(4), septic shock (5), endotoxemia (6)(7), cardiac transplant (8), hypertension (9), and cancer (10). Although the NO chemiluminescence methodology has been used to measure NO₂^- and NO₃^- (11)(12)(13), there are few reports evaluating the operating conditions of the assay.

The operating efficiency of the NO chemiluminescent system is governed by several variables: driving pressure and flow rate of the carrier gas (14)(15), which determine the degree of mixing in reducing solution and dispersion of NO in the carrier gas stream; chemiluminescent reaction chamber pressure (P_RC); and the selectivity (16), pH (11), temperature (13), and concentration of the reducing agents used to convert NO₂^- and NO₃^- to NO. Yang et al. (13) studied the efficiency of conversion of both NO₂^- and NO₃^- to NO using various reducing agents over a range of operating temperatures; however, no studies on the relationship between carrier gas flow rate and P_RC have been reported.

The present study had three primary objectives: (a) to determine the optimal operating conditions for carrier gas flow rate and P_RC to achieve maximum efficiency of the chemiluminescent response for both NO₂^- and NO₃^- determinations; (b) to determine the detection limit and linearity of NO₂^- and NO₃^- responses on the basis of these optimal operating conditions; and (c) to evaluate the recovery of NO₂^- and NO₃^-from plasma and deproteinized plasma under optimized . . . [Full Text of this Article]

Acknowledgments

References

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