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- Title
Nitric oxide isotope tracing at biological trace levels.
- Authors
Mills, Lauren; Feelisch, Martin; Grisham, Matthew B.; Rodriguez, Juan
- Abstract
The many roles of nitric oxide (NO) in biology and its tremendous therapeutic applications have drawn much attention to this molecule; yet little is still known on the chemical mechanisms that enable NO to achieve these roles in living systems. Here we describe new methodology, based on isotope labeling, for identifying and quantifying biochemical sources and targets of NO, in tissues and in biological fluids at biological trace levels. To detect 14NO and the stable isotope 15NO we have adapted a chemical assay, designed originally for chemiluminescence detection, to take advantage of mass spectrometric detection. Using a standard Sievers reaction vessel, coupled to an HP GCDI800A gas chromatograph / mass spectrometer via cryogenic focusing in a 1 ml loop, we were able to reach detection limits of 10 pmol and 50 pmol for 14NO and 15NO isotopes, respectively. The difference in detectability currently arises from oxygen contamination contributing to the background at 15NO, due to the limited mass selectivity of our instrument. A mass spectrometer upgrade, now under way, is expected to improve our sensitivity and selectivity manifold to where the limit of detection for both 14NO and 15NO isotopes should reach the single pmol range. At these levels our mass spectrometric approach should greatly expand the analytical capabilities available to investigators in the field of NO Chemical Biology.
- Subjects
ISOTOPES; NITRIC oxide; BIOLOGY; THERAPEUTICS; OXYGEN; MASS spectrometers
- Publication
FASEB Journal, 2007, Vol 21, Issue 5, pA623
- ISSN
0892-6638
- Publication type
Article
- DOI
10.1096/fasebj.21.5.a623-c