Determination of N-15-labeled ammonium and total nitrogen in plant and fungal systems using mass-spectrometry

A selected ion-monitoring method to measure 15N-labelled ammonia in biological samples was improved to simplify sample handling, to obviate interference from ammonia due to the decomposition of glutamine and to allow the determination of total N. Ammonia is derivatized with pentafluorobenzoylchloride to yield pentafluorobenzamide which is analysed by gas chromatography/mass spectrometry after clean-up using disposable silicic acid columns. The sensitivity achieved when operating in the negative ion-chemical ionization mode was somewhat higher than when using electron-impact ionization. Use of methyl amine as an internal standard improved the accuracy and precision of the measurements. The method was applied to samples taken from an intact ectomycorrhizal system fed with ISN-labelled ammonium and used to determine patterns of N assimilation into ammonium, free amino acids and macromolecular compounds