The impact of pulse duration on composite WATERGATE pulse

As an effective method for solvent suppression, WATERGATE is widely used in high resolution NMR spectroscopy. It is usually composed of a number of pulses separated by constant intervals. However, theoretical and experimental analyses indicate that narrower bandwidth and lower intensities around the secondary suppression points occur in the excitation profile of the composite WATERGATE. The excitation profile distortion is caused by the chemical shift evolution during the RF pulses. The higher the ratio of pulse duration to the inter-pulse delay is, the severer the profile distorts. Therefore, in high magnetic fields, the effect will be serious when WATERGATE is applied to some biological samples whose resonances distribute over a wide range. As can be seen obviously by applying WATERGATE to detect a RNA-protein mixture sample in an 800 MHz spectrometer, the resonances of the imino protons were partially suppressed by showing decreased intensities, though the intended secondary suppression points were set far away from them. In this article, we proposed an optimized WATERGATE that could effectively compensate the chemical shift evolution during the RF pulses, and relieve the excitation profile distortion. The optimized experiment will be a good way to retain the imino signal intensities when WATERGATE is applied to detect the RNA samples in high magnetic field.