HMQC/HSQC experiment

The other common inverse-detection method, heteronuclear multiple quan-turn coherence (HMQC) relies on multiple-quantum coherence transitions during the pulse sequence. Due to the multiple-quantum coherence transitions it is more laborious to theoretically follow the course of magnetization, and the cross peak will be broader in the Fi dimension due to the /hh evolution. Unlike HSQC, HMQC can also be optimized for Jch couplings. This heteronuclear multiple bond correlation experiment, or HMBC, ° ° has lower sensitivity than HMQC/HSQC experiments, and the Jch correlations can appear as artefacts in the spectrum. However, the cross peak volume should follow the concentration of analyte, so with proper method validation HMQC and HMBC should also be applicable for quantification. 

The 3D HMQC-NOESY-HMQC (HSQC) experiment together with 4D -edited... 

NOESY-HMQC (HSQC) experiment (b) 4D i C/i W-edited and i C/ C-edited NOESY experiments. Colour notation is identical with Fig. 5.17. 

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. 

The most powerful techniques of all are undoubtedly the 2-D proton-carbon experiments (Hetero-nuclear Multiple Quantum Coherence///eteronuclear Single Quantum Coherence, or HMQC/HSQC and //ctcronuclcar Multiple Bond Correlation, or HMBC) as they provide an opportunity to dovetail proton and carbon NMR data directly. 

The HSQC experiment is based on single rather than multiple quantum coherence during the evolution time, t. The contemporary multiplicity-edited gradient HSQC pulse sequence is shown in Fig. 10.15. Relative to the much simpler HMQC pulse sequence, the HSQC... 

Because of the favorable cross-peak multiplet fine-structure, the HSQC experiment offers superior spectral resolution over the HMQC (heteronuclear multiple quantum coherence) experiment [13, 14], On the other hand, the HMQC experiment works with fewer pulses and is thus less prone to pulse imperfections. The real advantage of the HSQC experiment is for measurements of samples at natural isotopic abundance and without the use of pulsed field gradients, since the HSQC experiment lends itself to purging with a spin-lock pulse. Spin-lock purging in the HMQC experiment... 

This suppression scheme has been shown to work well together with HMQC experiments of small molecules at natural abundance. Even cleaner spectra are obtained, if the BIRD sequence is combined with HSQC experiments already containing a spin-lock purge pulse. Drawbacks of the BIRD pulse scheme are the fact that the relaxation delay between scans cannot be chosen freely anymore and that complete suppression of all C-bound proton signals is impossible, if they have different relaxation times. Furthermore, the BIRD pulse scheme is not applicable to molecules in the slow motional regime, since negative NOEs between the inverted proton spins and the non-inverted C-bound proton spins would reduce the magnetization of the latter. 

Comparison of the results of the one-dimensional gradient supported 31P/15N 1H -se-HSQC experiment with phase-cycled HSQC and HMQC experiments gave relative S/N-ratios of 0.75 0.92 1 which was, under consideration of the suppression of one of the two possible coherence transfer pathways by the field gradients and the longer duration of the pulse sequence, interpreted in terms of a very good performance.25 The main benefits of the PFG-es-HSQC sequence were seen, however, in the excellent level of artefact suppression which allowed one to observe correlations via very small couplings even in cases where the active isotopomer is present in low natural abundance and its lines are normally obscured by residual parent signals (Fig. 3). 

In contrast to the basic "C detected experiment, and as a consequence of the final H detection, the 2D spectra obtained with HMQC or HSQC have a projection onto the F2 axis which corresponds to the normal H spectrum and includes all chemical shifts and all Jfi, couplings. The latter may give rise to rather broad cross peaks for extensively coupled protons. The projection onto the Fl axis corresponds to a normal C spectrum but with the quaternary carbons missing. With HMQC, but not with HSQC, cross peaks are additionally split in Fl by "J couplings. The HMQC and the HSQC experiment are usually performed in phase-sensitive mode, which, after proper phasing in both dimensions, allow peaks to be displayed in pure absorption. 

Arguably, the most important experiment for the study of humic materials is the HMQC or HSQC experiment. The HSQC or HMQC experiment detects... 

Later we will see how these couplings can be exploited in experiments that enhance the sensitivity of 13C spectra (INEPT), measure the number of hydrogens attached to each carbon (APT and DEPT), and correlate 13C chemical shifts with H chemical shifts using a second dimension (2D-HETCOR, -HMQC, -HSQC, and -HMBC). But for detecting a simple 13C spectrum, we need a way to suppress these 13C- H couplings so we can observe a single line (singlet) for each 13 C resonance. 

ID ll spectrum, and crosspeaks are arranged symmetrically around the diagonal. There is only one radio frequency channel in a homonuclear experiment, the H channel, so the center of the spectral window (set by the exact frequency of pulses and of the reference frequency in the receiver) is the same in If and F (Varian tof, Bruker ol). The spectral widths should be set to the same value in both dimensions, leading to a square data matrix. Heteronuclear experiments have no diagonal, and two separate radio frequency channels are used (transmitter for F2, decoupler for F ) with two independently set spectral windows (Varian tof and dof, sw, and swl, Bruker ol and o2, sw(If), and sw(I )). Heteronuclear experiments can be further subdivided into direct (HETCOR) and inverse (HSQC, HMQC, HMBC) experiments. Direct experiments detect the X nucleus (e.g., 13C) in the directly detected dimension (Ff) using a direct probe (13C coil on the inside, closest to the sample, H coil on the outside), and inverse experiments detect XH in the To dimension using an inverse probe (XH coil on the inside, 13C coil outside). 

Is the probe direct, or inverse The former is good for direct observation with or without INEPT enhancement. The latter will give poor signal-to-noise in direct experiments since the sample does not fill the coil space, but is much preferred for indirect detection via, for example, a heteronnclear mnltiple qnantum coherence (HMQC) or heteronnclear single qnantum coherence (HSQC) experiment. 

With suitable modification of the pulse sequences the para-hydrogen effect can be used to enhance the signals in many NMR experiments, for example, COSY, HMQC, HSQC, EXSY and Overhauser spectroscopy. ... 

Similar to the HSQC experiment, multiple quantum coherences can be used to correlate protons with Q-coupled heteronuclei. The information content of the Heteronuclear Multiple Quantum Correlation (HMQC) experiment (56) is equivalent to the HSQC, but the sensitivity can be improved in certain cases. Additionally, by proper tuning of delays and phase cycling, it can be transformed into the heteronuclear multiple bond correlation experiment (57-59), which results in correlations between J- and J-coupled nuclei. 

A very efficient suppression of parent resonances can be achieved using the T filter. This, however, requires a rather careful tuning of the relaxation delay T (see Figure 8). If the jump and return inversion pulse is employed, the pulse sequence can be regarded as a selective version of the BIRD experiment [57-59]. Obviously, multiple-frequency selective inversion pulses may be necessary in the case of complex proton spectra. Usually the /-BIRD HMQC experiment gives cleaner spectra as compared with equivalent heteronuclear singlequantum coherence (HSQC) experiments, presumably because of fewer 180° pulses which are frequently a source of various artefacts. 

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