Isotopic labeling, mixed

Both Jlic and Jun couplings have been observed by Sott et alP in the spectra of isotopically labelled mixed complexes formed by the lithium salt of acetonitrile (LiCH2CN) and the chiral lithium amides, Li-([Pg.168]

When a molecule adsorbs to a surface, it can remain intact or it may dissociate. Dissociative chemisorption is conmion for many types of molecules, particularly if all of the electrons in the molecule are tied up so that there are no electrons available for bonding to the surface without dissociation. Often, a molecule will dissociate upon adsorption, and then recombine and desorb intact when the sample is heated. In this case, dissociative chemisorption can be detected with TPD by employing isotopically labelled molecules. If mixing occurs during the adsorption/desorption sequence, it indicates that the mitial adsorption was dissociative. 

Mechanism I was ruled out by an isotopic labeling experiment. The mixed anhydride of salicylic acid and acetic acid is an intermediate if nucleophilic catalysis occurs by mechanism 1. This molecule is known to hydrolyze in water with about 25% incorporation of solvent water into the salicylic acid. 

An important group of analytical methods is based on measurements of the change in isotopic ratio when active and non-active isotopes are mixed. In the simplest case, a known amount w1 of labelled analyte of known specific activity at is added to the sample. After isotopic mixing has been established sufficient of the analyte is separated (nor normally 100%) to allow the new specific activity a2 to be measured. Measurements of activity and the amount of the analyte separated are thus required. Subsequently the amount w2 of analyte in the sample may be calculated from equation (10.17). 

The first mode of the high resolution C-NMR of adsorbed molecules was recently reviewed Q-3) and the NMR parameters were thoroughly discussed. In this work we emphasize the study of the state of adsorbed molecules, their mobility on the surface, the identification of the surface active sites in presence of adsorbed molecules and finally the study of catalytic transformations. As an illustration we report the study of 1- and 2-butene molecules adsorbed on zeolites and on mixed tin-antimony oxides (4>3). Another application of this technique consists in the in-situ identification of products when a complex reaction such as the conversion of methanol, of ethanol (6 7) or of ethylene (8) is run on a highly acidic and shape-selective zeolite. When the conversion of methanol-ethylene mixtures (9) is considered, isotopic labeling proves to be a powerful technique to discriminate between the possible reaction pathways of ethylene. 

Mass spectrometry. Reaction of OH to form an ion, HS04, which can be measured by mass spectrometry was first demonstrated by Eisele and Tanner (1991). Figure 11.45 is a schematic diagram of this approach (Tanner et al., 1997). Air is sampled through an inlet system described in detail by Eisele et al. (1997) and mixed with isotopically labeled 34SOz, forming H24S04 via reactions discussed in Chapter 8.C.2 ... 

In some applications like newborn screening and filter paper blood spots, the internal standard that is labeled cannot be mixed with blood. It can only be present in the extraction solvents. Therefore, only the extracted metabolites can be quantitatively measured. I have denoted a term called pseudo-isotope dilution to account for the differences between traditional isotope dilution and the technique commonly used in newborn screening by MS/MS. A special analysis is capable using this technique, however, in terms of an extraction efficiency experiment. With isotope-labeled standards you can perform an experiment whereby a traditional isotope-dilution technique (internal standard added to liquid blood and spotted) is compared to pseudo-isotope dilution techniques (internal standard is added to the extraction matrix). The ratio of the results of these two analysis (pseudo/traditional) is the extraction efficiency. 

Because the characterization of support-bound intermediates is difficult (see below), solid-phase reactions are most conveniently monitored by cleaving the intermediates from the support and analyzing them in solution. Depending on the loading, 5-20 mg of support will usually deliver sufficient material for analysis by HPLC, LC-MS, and NMR, and enable assessment of the outcome of a reaction. Analytical tools that are particularly well suited for the rapid analysis of small samples resulting from solid-phase synthesis include MALDI-TOF MS [3-5], ion-spray MS [6-8], and tandem MS [9]. MALDI-TOF MS can even be used to analyze the product cleaved from a single bead [5], and is therefore well suited to the identification of products synthesized by the mix-and-split method (Section 1.2). The analysis and quantification of small amounts of product can be further facilitated by using supports with two linkers, which enable either release of the desired product or release of the product covalently bound to a dye [10-13], to an isotopic label [11], or to a sensitizer for mass spectrometry [6,14,15] (e.g., product-linker-dye- analytical linker -Pol). 

The quantitative determination of amperozide in plasma using MALDI-MS was demonstrated (Jespersen et al., 1995). This study required addition of a stable-isotope-labeled amperozide as an internal standard and a typical liquid-liquid extraction procedure with dry down and reconstitution. The sample was then mixed with the matrix and analyzed. Linearity was achieved over a range of 2.5-40 [iM. Other examples have been demonstrated (Duncan et al., 1993) which also required the use of stable-isotope analogs as internal standards. These examples also utilized TOF instruments with nitrogen lasers. 

Fig. 10 Mixed isotope photoaffinity labeling strategy for determination of the labeling site. After binding and photocrosslinking the target enzyme with a 1 1 mixture of the light (Do) and heavy (D7) isotopically labeled A/BPs, the construct is purified and tryptically digested. LC-MS/MS analysis of the peptide pool allows discrimination between the labeled and unlabeled peptides. The modified fragment can easily be retrieved and identified by searching for the isotope signature...

Penfold et al. [62] have also used neutron reflectivity to study the adsorption (structure and composition) of the mixed anionic/nonionic surfactants of SDS and C12E6 at the hydrophilic silica-solution interface. This is rather different case to the cationic/nonionic mixtures, as the anionic SDS has no affinity for the anionic silica surface in the absence of the Ci2E6. The neutron reflectivity measurements, made by changing the isotopic labelling of the two surfactants and the solvent, show that SDS is coadsorbed at the interface in the presence of the Ci2E6 nonionic surfactant. The variations in the adsorbed amount, composition, and the structure of the adsorbed bilayer reflect the very different affinities of the two surfactants for the surface. This is shown in Fig. 7, where the adsorbed amount and composition is plotted as a function of the solution composition. 

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