Formation and metabolism of nitrosamines in vivo, monitored by 14N-stable isotope labelling.

Microsomal metabolism of N-nitrosodimethylamine entails release of molecular nitrogen; the extent is determined by 15N stable isotope labelling and mass-spectrometric isotope ratio measurements. Exhalation of labelled nitrogen by rats treated with 15N-dimethylamine and nitrite or 15N-nitrite alone indicates that nitrogen may arise from nitrite via two pathways: either directly from nitrosation of primary amines or from secondary and tertiary amines with subsequent enzymic N-demethylation. The overall yield of nitrosamine formation, N-demethylation and nitrogen-release represent about 0.3-6% or the administered dose of dimethylamine (1.1 mmol/kg), depending upon the dose of nitrite (0.55-2.2 mmol/kg). 15N-stable isotope labelling and mass-spectrometric isotope ratio measurements are powerful tools for assessment of endogenous nitrosamine formation from nitrite. One hundred nmol of labelled nitrogen are easily detectable in vivo; with further methodological refinement the limit of detection may be lowered by two orders of magnitude.

Amino acid determination by capillary gas chromatography on Chirasil-Val. Enantiomer labelling and nitrogen-selective detection.

Amino acid analysis by enantiomer labelling and capillary gas chromatography on Chirasil-Val is superior to conventional gas chromatography and ion-exchange chromatography with respect to sensitivity, accuracy and speed. Employment of an alkali flame-ionization detector allows the selective detection of amino acids and suppression of background peaks; in addition, the detectability of amino acids is enhanced. Most nitrogen-selective detectors require meticulous adjustment of the operating conditions, but in combination with enantiomer labelling this is less critical. Maximum sensitivity and selectivity of the alkali-bead flame-ionization detector is achieved with a minimal flow of hydrogen. When using it as the carrier gas in capillary gas chromatography, flow control instead of the common pressure regulation is recommended to avoid a continuous fall of the baseline during temperature programming. Accurate flow control is achieved with a micro-aperture. The benefit of nitrogen-selective detection is especially apparent for histidine and arginine.