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Old 08-21-2010, 08:15 PM
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Default Watergate

WATERGATE

WATER suppression by GrAdient Tailored Excitation (WATERGATE) is a clever technique used to suppress the water signal in an aqueous sample. It is widely used in many complicated pulse sequences. Unlike presaturation which irradiates the water resonance with a long low power pulse, this method is based on the gradient spin echo technique used also to measure diffusion constants and DOSY spectra. The pulse sequence is shown here:The transmitter frequency is set on the water resonance. A non-selective hard 90° pulse is applied followed by a 1 -2 msec gradient pulse. The gradient pulse dephases all of the resonances. A composite pulse (consisting of 6 hard pulses seperated by a delay, ?) is then applied which acts as a 180° pulse for everything except peaks on resonance (i.e. water) and any peaks at frequencies n/? away from the transmitter, where n is an integer. ? is chosen such that 1/? lies outside of the spectral width (typically several hundred µsec). The second gradient pulse (equal in magnitude, duration and sign, to the first) further dephases the water resonance at the center of the spectrum which was unaffected by the composite pulse but rephases everything else which was inverted by the composite pulse. The gradients and composite pulse act as a gradient spin echo for all but the water. The FID is then collected with the water resonance suppressed by the two dephasing gradients. An example of the application of WATERGATE is shown in the figure below. The top trace shows a standard 500 MHz 1H NMR spectrum of phenylalanine in H2O / D2O scaled to the water peak. The resonances of the phenylalanine are not visible on this scale. The middle trace is the same spectrum as the top trace with the phenylalanine resonances on scale. The huge water resonance is truncated. The bottom trace shows the WATERGATE spectrum. The water signal is greatly suppressed.



Source: University of Ottawa NMR Facility Blog
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[U. of Ottawa NMR Facility Blog] Watergate vs Presaturation
Watergate vs Presaturation Biochemists and protein chemists are often interested in observing the NH protons in their samples. Since the NH protons usually undergo slow chemical exchange with water, it is desirable to run the samples in H2O rather than D2O so the NH protons will not exchange with the deuterium in the solvent which would make them invisible in the 1H NMR spectrum. In practice, a mixture of 10% D2O and 90% H2O is used as a solvent so that a deuterium lock can be established and used while running the spectrum. The very high concentration of water compared to the very low...
nmrlearner News from NMR blogs 0 08-21-2010 08:15 PM



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