Doubly isotopes

Fig. 8. An overview of the zero order energy levels of the doubly isotopically tagged helix. The isotope shifts are Sa (13C=I80) and Sb (l3C=l80) while Aa and Ab are diagonal anharmonicities. The shaded areas represent the helix one- and two-exciton bands that become perturbed by the isotopes. The solid (bra evolution) and dashed (ket evolution) arrows represent one of the Liouville paths contributing to the echo.

Willadsen and Eggerer (75) have studied the stereochemistry of the enzyme acetyl CoA acetyltransferase, a key enzyme in both the terminal step in C-3 oxidation of fatty acids and the initial step in the biosynthesis of terpenes and steroids. The enzyme, when incubated separately with (2S)-[2-2Hi,2-3Hi]aceto-acetyl CoA and the (2R) isomer gave two moles of acetyl CoA as depicted in Scheme 17. Eggerer et al. (76) utilized the enzyme enoyl CoA hydratase to convert properly labeled crotonyl CoA, via syn addition, to the doubly isotopically labeled 3-hydroxyacyl CoA derivatives needed in this study. A discussion of this unique type of hydration has been presented by Rose (9). The labeled... 

The oxidations of formic acid by Co(III) and V(V) are straightforward, being first-order with respect to both oxidant and substrate and acid-inverse and slightly acid-catalysed respectively. The primary kinetic isotope effects are l.Sj (25°C)forCo(IU)and4.1 (61.5 C°)for V(V). The low value for Co(lII) is analogous to those for Co(IIl) oxidations of secondary alcohols, formaldehyde and m-nitrobenzaldehyde vide supra). A djo/ h20 for the Co(III) oxidation is about 1.0, which is curiously high for an acid-inverse reaction . The mechanisms clearly parallel those for oxidation of alcohols (p. 376) where Rj and R2 become doubly bonded oxygen. 

The structures of doubly bonded compounds containing tin readily lend themselves to study by multinuclear NMR spectroscopy, as tin has two NMR active isotopes ll7Sn and ll9Sn. The chemical shifts of doubly bonded tin species in the 119Sn NMR spectrum cover a broad range from 6 = -150 to +835 (Table VI) however, the majority of the signals occur at low field S = 400 or above. 

A special, isotope-labeled case of the azide-tetrazole equilibrium was studied by Cmoch et al. <2000JP0480>, and the results are shown in Scheme 23. 2-Chloro-3-nitropyridine 86 was treated with potassium azide containing a doubly labeled (15NN15N) azide anion. The authors detected formation of two differently labeled tetrazolopyridines the 2,4- 87 and the 1,3-labeled 88 derivatives. 

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. 

The reaction was second order in acid and first order in substrate, so both rearrangements and the disproportionation reaction proceed via the doubly-protonated hydrazobenzene intermediate formed in a rapid pre-equilibrium step. The nitrogen and carbon-13 kinetic isotope effects were measured to learn whether the slow step of each reaction was concerted or stepwise. The nitrogen and carbon-13 kinetic isotope effects were measured using whole-molecule isotope ratio mass spectrometry of the trifluoroacetyl derivatives of the amine products and by isotope ratio mass spectrometry on the nitrogen and carbon dioxide gases produced from the products. The carbon-12/carbon-14 isotope... 

The isotope effects for transfer of hydrogen were 1.79 for transfer from OH to N and 2.86 for transfer from CH to ruthenium. The isotope effect for transfer of the doubly labelled material d% 2-propanol) was 4.88, within the experimental error. If the hydrogen atoms would be transferred in separate... 

Although the multiple isotope method is most frequently used with stable isotopes (for example studies of oxygen KIE s in biophosphates used 1SN at a remote nitro group, or 13C on a remote carboxy group, as reporting isotopes), the technique is not restricted to stable isotopes radioisotopes have been used as reporting sites for stable isotopes. In a practical sense this is the only method that allows the measurement of isotope effects for elements that have only one stable isotope (e.g. fluorine and phosphorus). In these cases doubly radiolabeled material is used (see Section 7.4). 

Figure 5. Normal modes for vibration of tetrahedral [Cr04] (chromate). There are four distinct vibrational frequencies, including one doubly-degenerate vibration (E symmetry) and two triply-degenerate vibrations (F2 symmetry), for a total of nine vibrational modes. Arrows show the characteristic motions of each atom during vibration, and the length of each arrow is proportional to the magnitude of atomic motion. Only F2 modes involve motion of the central chromium atom, and as a result their vibrational frequencies are affected by Cr-isotope substitution. The normal modes shown here were calculated with an ab initio quantum mechanical model, using hybrid Hartree-Fock/Density Functional Theory (B3LYP) and the 6-31G(d) basis set—other ab initio and empirical force-field models give very similar results.

Traditionally thermal ionization mass spectrometry was the instrument of choice for the isotopic analysis of metals because thermal ionization produced an ion beam with a very small kinetic energy spread ( 0.5 eV). Therefore only a magnetic mass analyzer is needed to resolve one isotope from another. Moreover, ionization of unwanted material, such as atmospheric contaminates, hydrocarbons from pump oil, or production of doubly ionized particles is almost non existent, thus background counts are minimized and signal-to-noise ratio is maximized. 

Doubly-charged ions exist because the potential of second ionization of many metals is relatively low with respect to the plasma thermal energy. For instance, 11.9 eV are needed to remove two electrons, in contrast with 6.1 eV for one electron, from a calcium atom. At 8000K, a little less than 0.1% of Ca would be in the Ca + form and overlap with Mg+ isotopes. Likewise, Ba is rather easily formed and overlaps with Zn+ isotopes. Isobaric interferences with doubly-charged ions are easily identified as odd-mass atoms will produce peaks at half masses, such as Ca at mass 21.5. 

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