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The selective ge-1D HSQC-NOESY experiment allows to obtain high-quality 1D HSQC-NOESY spectra from which homonuclear 1H-1H NOEs can be elucidated starting from a selected carbon. Thus, in the first part of this experiment, magnetization is transferred from the selected carbon to the directly-bonded protons via a selective HSQC block. After this, the in-phase magnetization of these protons is allowed to evolve by a conventional NOESY block under the dipolar coupling effect. The same principles described here are also applied to the selective ge-1D HMQC-NOESY experiment.REQUIREMENTS
Easy implementation on AVANCE spectrometers equipped with pulsed field gradients, selective excitation using shaped pulses and inverse probehead.VERSIONS
The basic pulse sequence of the selective ge-1D HSQC-NOESY experiment is exactly the same as the conventional ge-2D HSQC-NOESY pulse train in which the following modifications have been included ( 95JMRA32-114 and 97MRC199 ):EXPERIMENTAL DETAILS
- One of the two 90º 13C pulses in the HSQC block is made selective on a specific resonance. In order to improve the selectivity, proton decoupling is simultaneously applied during the long selective 13C pulse.
- The variable evolution period of the 2D version is fixed to a minimum delay (3 microseconds).
- Gradients are usually applied for coherence selection purposes in natural abundance samples to obtain efficient suppression of undesired 1H-12C magnetization. In theory, half of the signal is lost with compared to conventional phase-cycled experiment.
- Carbon decoupling during proton acquisition is optional. In the coupled version, the two satellites of the directly-attached protons will appear as in-phase multiplet. In the decoupled version, these satellites collapse in a single signal.
The selective ge-1D HSQC-NOESY experiment can be run with minor changes from a predefined parameter set. The HSQC block is optimized as discussed in the selective ge-1D HSQC experiment and the NOESY mixing time is optimized as usual. Important parameters to consider are:SPECTRASelectivity of the selective excitation 13C pulse: the user must define the offset, the shape, the duration and the power level needed for a defined excitation profile. Optimization of the J-coupling delay in the HSQC block as a function of 1/(4*J(CH)), in order to get in-phase magnetization of the directly-bonded protons. Optimization of the NOESY mixing period as a function of the sample under study.
The selective ge-1D HSQC-NOESY experiment affords a simple 1D 13C-edited NOESY spectrum in which the protons directly-bonded to the selected carbon appear as a large doublet due to 1JCH and the protons J-coupled with them appear as in-phase multiplets. The use of gradients allows to obtain a clean, artefact-free spectrum in a short time in which perfect suppression of undesired 1H-12C magnetization is achieved with a single two-step phase cycle (the selective 90º carbon pulse and the receiver are usually inverted on alternated scans).RELATED TOPICS
Selective ge-1D HSQC-ROESY experiment
Selective ge-1D HMQC-NOESY experiment
Selective ge-1D HMQC-ROESY experiment