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Updated: Aug 29, 2026

Analytical Techniques for Assaying Nitric Oxide Bioactivity
Published on: June 18, 2012
A gas chromatographic method for the indirect determination of hydroxylamine in pharmaceutical preparations:
J P Guzowski1, C Golanoski, E R Montgomery
1Bristol-Myers Squibb, Pharmaceutical Research Institute, 1 Squibb Drive, New Brunswick, NJ 08903, USA. john.guzowski@bms.com
Abstract:
A simple, sensitive, and selective headspace gas-chromatographic method has been developed for measuring hydroxylamine (HA) in a variety of sample matrices including pharmaceutical formulations. This procedure relies on converting HA into nitrous oxide (N2O), which is a single-step reaction that is carried out directly in a heated headspace vial. The gaseous products are then analyzed by headspace capillary gas chromatography. Several detection strategies were evaluated and electron capture provided the best sensitivity (4 parts-per-billion (ppb)) while the mass selective and thermal conductivity values were higher (14 ppb and 1.4 parts-per-million (ppm), respectively). The method's linear dynamic range spans two to four decades with a run-to-run precision that was better than 5% R.S.D. (n=7). The reagent concentrations (oxidant, buffer) strongly impact the N2O signal and the greatest response was obtained for solutions that contained equimolar amounts of reactants. HA was efficiently (98%) recovered from a sample matrix that contained only the active pharmaceutical ingredient (API) but the recovery was lower (83%) when excipients were present.
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