Nitrogen isotopic labelling

The mechanism outlined above is supported by experimental findings. An intermediate 5 has been isolated, " and has been identified by and N-nuclear magnetic resonance spectroscopy. Side-products have been isolated, which are likely to be formed from intermediate 4. N-isotope labeling experiments have shown that only the nitrogen remote from the phenyl group is eliminated as ammonia. 

What was the solution The reaction had to be carried out with a diazonium ion labeled with one or two atoms of the nitrogen isotope 15N in the presence of large concentrations of unlabeled nitrogen molecules (14N2), quenched before completion, and the remaining diazonium ion analyzed for 15N content if this had decreased relative to the initial value, the dogma would be falsified. 

Thus, upon photolyzing a mixture of two nitrites, one of which contains an isotopically labeled nitrogen, the products should show the original isotopic distribution if mechanism (a) is operative, the original isotopic distribution in unreacted nitrite but scrambling in the nitroso product if (c) is operative, and complete isotopic scrambling if (b) is operative. 

Although use of radio and stable isotope labels involving the trio of covalently-bonded nitrogenous functions in 3 and in 78, provided evidence that isocyano is the precursor of the isothiocyano and formamido groups [30, 81], it remains to be shown that a biosynthetic equivalent of the in vitro chemically-proven fusion process between isocyano and free sulfur (e.g., cf. Introduction) exists in the cells of sponges. In marine biota, various ionic forms of sulfur in a number of oxidation states, as well as organo-polysulfides are known. However, any association with the isonitrile group and a sulfated species has yet to be established. 

In addition to the foregoing examples, periodate oxidation has been applied to isotopically labeled sugars (for the determination of label distribution), 226 -226 to certain branched-chain sugars,227-229 and to some nitrogenous derivatives of the simple sugars.230-236... 

The 2,2 -hydrazonaphthalene doubly labelled with nitrogen-15 for the nitrogen isotope effect experiments and the [l,l -13C2]-2,2 -hydrazonaphthalene required for measuring the carbon isotope effect were synthesized by the reaction sequence shown in Schemes 18 and 19. 

The nitrogen kinetic isotope effect of 1.0197 found using the substrate with the natural abundance of nitrogen isotopes corresponds to an isotope effect of 1.04 for the reaction of the doubly labelled compound. Thus, the nitrogen isotope effects found using two different analytical techniques to measure the isotope effect are in excellent agreement. 

The doubly nitrogen-15 labelled substrate required for determining the nitrogen isotope effect for this reaction was obtained by the reactions shown in Scheme 2340. The series of reactions used in the synthesis of the [4,4 -13C2]-4,4 -dichlorohyrazobenzene is shown in Scheme 2440, and the preparation of the [2-14C]- and the [4-14C]-4,4 -dichlorohyrazobenzene are described in Schemes 25 and 26. 

Work on the molecular structure of benzonitrilio methylide 8/9 has been carried out via Fourier transform infrared (FTIR) smdies on it and five isotope-labeled variants. The nitrile ylides were generated in nitrogen matrices at 12 K either directly, by photolysis of the azirine 7, or indirectly from the azidostyrenes 6 (4). 

Acylcarnitine analysis of dried blood or bile spots is very similar to the analysis of plasma. A small disk (diameter typically 5 mm or less) is punched out of the blood spot and the acylcarnitines extracted by the addition of methanol and known concentrations of isotopically labeled acylcarnitines, which function as internal standards. The extract is dried under a stream of nitrogen, and derivatized by the addition of either n-butanol HC1 or n-methanol HC1. The acylcarnitines are measured as their butyl or methyl esters by MS-MS. The concentrations of the analytes are established by computerized comparison of ion intensities of these analytes to that of the internal standards. 

Back and Mui17 studied the reaction with isotopically labeled nitric oxide to test the proposal that active nitrogen contained vibra-tionally excited molecules capable of dissociating nitric oxide. This reaction was proposed by Winkler and his co-workers133,435... 

The precise nature of the reaction path is ambiguous, because the same products would be obtained regardless of whether an oxygen atom or a chlorine atom were transferred. As they pointed out in their paper, this ambiguity can be resolved by using nitryl chloride with isotopically labeled nitrogen. Pertinent to this question are the calculations of Herschbach, Johnston, Pitzer, and Powell,199 who computed pre-exponential factors based on two alternative activated complexes... 

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