Multiple quantum detection

A H(detected)- C shift correlation spectrum (conmion acronym HMQC, for heteronuclear multiple quantum coherence, but sometimes also called COSY) is a rapid way to assign peaks from protonated carbons, once the hydrogen peaks are identified. With changes in pulse timings, this can also become the HMBC (l eteronuclear multiple bond coimectivity) experiment, where the correlations are made via the... 

An alternative way of acquiring the data is to observe the signal. These experiments are referred to as reverse- or inverse-detected experiments, in particular the inverse HETCOR experiment is referred to as a heteronuclear multiple quantum coherence (HMQC) spectmm. The ampHtude of the H nuclei is modulated by the coupled frequencies of the C nuclei in the evolution time. The principal difficulty with this experiment is that the C nuclei must be decoupled from the H spectmm. Techniques used to do this are called GARP and WALTZ sequences. The information is the same as that of the standard HETCOR except that the F and F axes have been switched. The obvious advantage to this experiment is the significant increase in sensitivity that occurs by observing H rather than C. 

HC HMQC (heteronuclear multiple quantum coherence) and HC HSQC (heteronuclear single quantum coherence) are the acronyms of the pulse sequences used for inverse carbon-proton shift correlations. These sensitive inverse experiments detect one-bond carbon-proton connectivities within some minutes instead of some hours as required for CH COSY as demonstrated by an HC HSQC experiment with a-pinene in Fig. 2.15. 

HMQC Heteronuclear multiple quantum coherence, e.g. inverse CH correlation via one-bond carbon proton-coupling, same format and information as described for ( C detected) CH COSY but much more sensitive (therefore less time-consuming) because of H detection... 

Phase cycling is widely employed in multipulse NMR experiments. It is also required in quadrature detection. Phase cycling is used to prevent the introduction of constant voltage generated by the electronics into the signal of the sample, to suppress artifact peaks, to correct pulse imperfections, and to select particular responses in 2D or multiple-quantum spectra. 

H-Detected Heteronuclear Multiple-Quantum Coherence (HMQC) Spectra... 

HMQC //ctcronuclear multiple quantum coherence. A proton-detected, 2-D technique that correlates protons to the carbons they are directly attached to. 

The first of the proton-detected experiments is the Heteronuclear Multiple Quantum Correlation HMQC experiment of Bax, Griffey and Hawkins reported in 1983, which was first demonstrated using 1H-15N heteronuclear shift correlation [42]. The version that has come into wide-spread usage, particularly among the natural products community, is that of Bax and Subramanian reported in 1986 [43]. A more contemporary gradient-enhanced version of the experiment is shown in Fig. 10.14 [44],... 

Fig. 10.14. Gradient-enhanced HMQC pulse sequence described in 1991 by Hurd and John derived from the earlier non-gradient experiment of Bax and Subramanian. For 1H-13C heteronuclear shift correlation, the gradient ratio, G1 G2 G3 should be 2 2 1 or a comparable ratio. The pulses sequence creates heteronuclear multiple quantum of orders zero and two with the application of the 90° 13C pulse. The multiple quantum coherence evolves during the first half of ti. The 180° proton pulse midway through the evolution period decouples proton chemical shift evolution and interchanges the zero and double quantum coherence terms. Antiphase proton magnetization is created by the second 90° 13C pulse that is refocused during the interval A prior to detection and the application of broadband X-decoupling.

This scheme is applied in the so-called X-half-filter technique (Fig. 17.4a, c), with the only difference of an additional 90 (X) pulse with constant phase [16, 17]. Instead of generating heteronuclear multiple quantum coherence 2 Iy Sy, which cannot readily be detected (in case of selecting the 1H-X pairs), one now always ends up with proton antiphase coherence, but with the same phase alternation ... 

Inverse-detected experiments have had the greatest effect in making 15N NMR experiments feasible for small samples. These experiments take advantage of the higher sensitivity of NMR to facilitate the observation of insensitive nuclei like 13C and 15N. The H-13C heteronuclear multiple quantum coherence (HMQC) and the related heteronuclear multiple-bond correlation (HMBC) experiments are important in contemporary natural products... 

HMBC ( -Detected Multiple-bond Heteronuclear Multiple Quantum Coherence Spectrum) NMR of 26a allowed one to show that the isopropyl group (Ha, Hfc) is connected to the olefmic carbon (Cc), whereas methyl (IF) and methylene (H-C, H- ) groups are bonded to another olefmic carbon (Cd) and fullerene carbon (Ch) through quaternary carbon (Ce) shown in 26a and 26b (Scheme 8). 

Heterocorrelations can be detected both in direct and reverse modes. In the latter mode, dramatic enhancements of sensitivity can be achieved owing to the larger sensitivity of protons with respect to heteronuclei. In the most common heterocorrelation pulse sequences for reverse detection, called heteronuclear multiple quantum coherence (HMQC) (Fig. 8.2G) [25,26], H-I3C MQ (multiple quantum) coherence is generated by first applying a 90° pulse on protons and, after a time t chosen equal to 1/2 J[j, by applying a 90° pulse on carbon (Fig. 8.19). 

Fig. 8.19. Vector representation of a H-13C HMQC experiment. The first 90° pulse along y rotates the equilibrium magnetization of the proton spin, /H, from the z axis to the x axis. After a time /d = 1/2/Hx, the antiphase coherence 2/J1/ t (see Appendix IX) is at its maximum. A 90° pulse on carbon along y then transforms the antiphase coherence into a MQ (multiple quantum) coherence (the 2/J1/ component is shown). During t the MQ evolves (with a 180 refocusing pulse on proton in the middle), until a further 90 pulse on carbon along x transforms the —2/ / component (shown at its maximum for clarity) into a 2/ /f antiphase coherence. After the time fd, in-phase coherence of the proton spin develops. The latter is detected during h. Its initial intensity is modulated by the carbon Larmor frequency during t (if proton refocusing has been used), thus originating a proton-carbon cross peak.

In order to carry out complete structural elucidation of unknown compounds (especially for complex molecules), the RF probe should enable a variety of heteronuclear NMR techniques to be performed. In particular, inverse detection H-15N and 1H-13C experiments such as heteronuclear multiple quantum coherence (HMQC) [29,30] and heteronuclear single quantum coherence (HSQC) [31] find almost ubiquitous application in myriad research environments. Although the microliter-scale probes described above feature both heteronuclear and homonuclear capabilities, no commerical product is... 

Great progress was made by Marzilli143 on using -detected multiple-quantum NMR, in the structure determination of d(TGGT). 

Figure 4. Two-dimensional heteronuclear multiple quantum coherence spectrum of a medium sample taken from a culture of HeLa cells which had been grown for 48 hours in the presence of 2 mM L[2-15N]glutamine. In this type of experiment the, 5N label (FI axis) is detected indirectly via spin-coupled protons (F2 axis). The peaks labeled in the contour plot arise from alanine (Ala), glutamate (Clu), glutamine (Gin), glycine (Gly), aspartate (Asp), and pyrollidone carboxylic acid (Pyr). From Street etal., 1993, with permission.

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