Nuclear magnetic resonance spectroscopy limitations

Nuclear Magnetic Resonance Spectroscopy. Bmker s database, designed for use with its spectrophotometers, contains 20,000 C-nmr and H-nmr, as weU as a combined nmr-ms database (66). Sadder Laboratories markets a PC-based system that can search its coUection of 30,000 C-nmr spectra by substmcture as weU as by peak assignments and by fiiU spectmm (64). Other databases include one by Varian and a CD-ROM system containing polymer spectra produced by Tsukuba University, Japan. CSEARCH, a system developed at the University of Vieima by Robien, searches a database of almost 16,000 C-nmr. Molecular Design Limited (MDL) has adapted the Robien database to be searched in the MACCS and ISIS graphical display and search environment (63). Projects are under way to link the MDL system with the Sadder Hbrary and its unique search capabiHties. 

GM Clore, MA Robien, AM Gronenborn. Exploring the limits of precision and accuracy of protein structures determined by nuclear magnetic resonance spectroscopy. J Mol Biol 231 82-102, 1993. 

Knight CTG, SD Kimade (1999) Silicon-29 nuclear magnetic resonance spectroscopy detection limits. A a/ Chem 71 265-267. 

Currently, there are no accurate methods available for quantifying the aliphatic bridges in the coal macromolecule. Quantitative nature of the application of infrared (IR) spectroscopy is limited to certain general types of functional groups or bond types. Nuclear magnetic resonance spectroscopy, despite the success of dipolar dephasing techniques to decipher the extent of substitution on carbon atoms, is still inadequate to distinguish distinct structural entities . 

Infrared spectroscopy has been used for quantitatively measuring the amounts of 1,2-, 3,4-, cis-1,4-, and trans-1,4-polymers in the polymerization of 1,3-dienes its use for analysis of isotactic and syndiotactic polymer structures is very limited [Coleman et al., 1978 Tosi and Ciampelli, 1973]. Nuclear magnetic resonance spectroscopy is the most powerful tool for detecting both types of stereoisomerism in polymers. High-resolution proton NMR and especially 13C NMR allow one to obtain considerable detail about the sequence distribution of stereoisomeric units within the polymer chain [Bovey, 1972, 1982 Bovey and Mirau, 1996 Tonelli, 1989 Zambelli and Gatti, 1978],... 

Nuclear magnetic resonance spectroscopy can detect the presence of aldehydo and keto forms of sugars in those rare instances where they occur to the extent of 1% or more in equilibrium, their proportion has thus been determined.16,20,23 24 However, the percentage of the acyclic forms present in equilibrium is usually very small, and is much below the limit of detection by n.m.r. spectroscopy other methods have, therefore, to be used. 

Fig. 3. Calculated NMR lineshapes for equally populated two-site exchange as a function of the dimensionless parameter a = nf rA. The abscissa is the dimensionless relative offset parameter, x = A/// (see Eq. (18)). (a) a = 4 (b) a = 2 (c)a=l (d) a = l/ /2 (e) a = 0.5 (f) a = 0.2. Spectra (a) and (f) are near the slow and fast exchange limits, respectively. Reproduced with permission from R. K. Harris, Nuclear Magnetic Resonance Spectroscopy A Physicochemical View, p. 124, Longman Scientific and Technical, Harlow, 1986.

Most of our structural information comes from x-ray crystallographic analysis of protein crystals and from the use of nuclear magnetic resonance spectroscopy in solution. Each of these techniques has advantages and limitations which makes them suitable for a complementary range of problems. The first protein structure determined at a sufficient resolution to trace the path of the polypeptide chain was that of myoglobin in 1960. Since that time many thousands of structures corresponding to hundreds of different proteins have been determined. The coordinates of the atoms in many protein and nucleic acid structures are available from the Protein Data Bank, which may be accessed via the Internet or World Wide Web (http //www.pdb.bnl.gov). 

Hanna, J.V., Smith, M.E., Stuart, S.N. Healy, P.C. (1992)7. Rhys. Chem., 96, 7560. Harris, R.K. (1983) Nuclear Magnetic Resonance Spectroscopy, Pitman Books Limited, London. 

Nuclear magnetic resonance spectroscopy (NMR) provides reasonably detailed structural information on a molecule and is an extremely useful method for characterization of impurities (see discussions in Section VII and in Chapter 12) however, its use as a quantitative method is limited. 

The structure and identity of such compounds that are of practical relevance as com-plexing agents may be elucidated unequivocally by both one-dimensional and two-dimensional nuclear magnetic resonance spectroscopy of the isotopes H-l, C-13, and P-31. Sufficiently high concentrations also render possible their quantitative analysis [87-91]. However, because of the low sensitivity, especially of the phosphorus nucleus, problems are encountered with the limits of detection in practical applications. 

Lithium metabolism and transport cannot be studied directly, because the lack of useful radioisotopes has limited the metabolic information available. Lithium has five isotopes, three of which have extremely short half lives (0.8,0.2, 10 s). Lithium occurs naturally as a mixture of the two stable isotopes Li (95.58%) and Li (7.42%), which may be determined using Atomic Absorption Spectroscopy, Nuclear Magnetic Resonance Spectroscopy, or Neutron Activation analysis. Under normal circumstances it is impossible to identify isotopes by using AAS, because the spectral resolution of the spectrometer is inadequate. We have previously reported the use of ISAAS in the determination of lithium pharmacokinetics. Briefly, the shift in the spectrum from Li to Li is 0.015 nm which is identical to the separation of the two lines of the spectrum. Thus, the spectrum of natural lithium is a triplet. By measuring the light absorbed from hollow cathode lamps of each lithium isotope, a series of calibration curves is constructed, and the proportion of each isotope in the sample is determined by solution of the appropriate exponential equation. By using a dual-channel atomic absorption spectrometer, the two isotopes may be determined simultaneously. - ... 

Nuclear Magnetic Resonance Spectroscopy. Like IR spectroscopy, NMR spectroscopy requires little sample preparation, and provides extremely detailed information on the composition of many resins. The only limitation is that the sample must be soluble in a deuterated solvent (e.g., deuterated chloroform, tetrahydro-furan, dimethylformamide). Commercial pulse Fourier transform NMR spectrometers with superconducting magnets (field strength 4-14 Tesla) allow routine measurement of high-resolution H- and C-NMR spectra. Two-dimensional NMR techniques and other multipulse techniques (e.g., distortionless enhancement of polarization transfer, DEPT) can also be used [10.16]. These methods are employed to analyze complicated structures. C-NMR spectroscopy is particularly suitable for the qualitative analysis of individual resins in binders, quantiative evaluations are more readily obtained by H-NMR spectroscopy. Comprehensive information on NMR measurements and the assignment of the resonance lines are given in the literature, e.g., for branched polyesters [10.17], alkyd resins [10.18], polyacrylates [10.19], polyurethane elastomers [10.20], fatty acids [10.21], cycloaliphatic diisocyanates [10.22], and epoxy resins [10.23]. 

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