JOURNAL OF MAGNETIC RESONANCE, Series A 123, 119121 (1996)
ARTICLE NO. 0222
Gradient-Selected NOESYA Fourfold Reduction of the Measurement
Time for the NOESY Experiment
RONALD WAGNER
Department of Chemistry, Philipps University Marburg, Hans-Meerwein-Strasse, D-35032 Marburg, Germany
Received July 29, 1996
The NOESY experiment (1, 2) is one of the fundamental
and most often performed 2D NMR experiments for applica-
tions in organic chemistry and in biomolecular research. In the
latter application, overnight and even longer measurements are
often required due to the low concentration of proteins or oligo-
nucleotides. A NOESY experiment for application in organic
chemistry to determine stereochemical assignments typically
runs about two hours, depending on the relaxation and mixing
time (3). This rather long time is not due to the sensitivity of
current NMR instruments but because an eight-step phase cycle
(4) is required to suppress unwanted coherences, such as dou-
ble-quantum contributions, COSY-like signals, and axial peaks.
In recent years, the other standard sequences used in structural
elucidation for organic chemistry, such as COSY (5) or HMQC
(6) have experienced a significant reduction in experiment time
with the introduction of pulsed field gradients (7). Today a
gradient-selected COSY experiment or a HMQC-type inverse
C,H correlation of typically 2050 mg of organic material and
a molecular weight on the order of 500 Da can be performed
in 5 to 10 minutes (3). Compared with this, the corresponding
NOESY measurement time is inconvenient, especially since it
is desirable to perform more than one NOESY experiment using
different mixing times. Within this context, Kock and Griesinger
recently published a FAST NOESY method (8), reducing the
relaxation waiting time. However, their procedure requires an
extra measurement of these relaxation times and special data
processing so that this method does not seem to have gained
wide application in service laboratories.
In this Communication, we wish to demonstrate that by an
extremely easy modification of the standard experiment, the
usual eight-step phase cycle of the experiment can effectively
be reduced to two steps leading to the desired fourfold reduction
in experiment time. We replace the NOESY mixing time tm
simply by a weak pulsed field gradient, which acts during the
entire exchange period as shown in Fig. 1.
For a two-spin system, it can be shown with the product-
operator formalism (9) that the second 907 pulse of the NOESY
sequence creates the desired frequency-labeled z magnetization,
but in addition double- and zero-quantum coherences and anti-
phase coherences leading to COSY-type signals. Despite the
* To whom correspondence should be addressed.
AID JMRA 0982 / 6j13$$$381 10-17-96 15:47:35
AND STEFAN BERGER *
zero-quantum coherences, these can be efficiently suppressed by
the pulsed field gradient in the mixing time, whereas the field
gradient will not affect the frequency-labeled z magnetization
which is the working principle of the NOESY scheme. In addi-
tion, axial peaks which are caused by spins which have relaxed
during t1 and which are made transverse again by the second
907 pulse are also suppressed.
The idea of placing a pulsed field gradient within the mixing
period seems to be straightforward and has indeed been used
in more complicated 3D sequences (10) and in selective 1D
versions of NOE difference spectroscopy (11). It was even
mentioned before the introduction of self-shielded gradient
coils in the first demonstration of the EXSY experiment (12)
and applied for the HOESY sequence (13). Surprisingly, to
the best of our knowledge, it has not been mentioned or demon-
strated for the basic homonuclear 2D NOESY sequence, proba-
bly because the developers of modern NMR need many tran-
sients for the usual protein work. Best results were achieved
when the gradient pulse was on during the entire mixing time.
This is probably due to the integrated gradient strength which
can be reached and to the suppression of stray-field effects
during the mixing time (14). In principle, our method could
be performed using no phase cycle at all; however, in practice,
it seems to be advantageous to use a two-step phase cycle,
similar to many other gradient-selected experiments (3, 7).
As an example, we show in Fig. 2a a standard NOESY
spectrum of a 10% solution of strychnine
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120 COMMUNICATIONS

FIG. 1. Pulse sequence for the gs-NOESY method. w1 , w2 , w3 , phases for the 907 1H NMR RF pulses; w4 , receiver phase; t1 , time increment for
the indirect dimension; tm , NOE mixing time; gz , pulsed field gradient.

in CDCl3 using the eight-step phase cycle, a relaxation time of
2 s, and a mixing time of 250 ms requiring an experiment time
of 83 min. Figure 2b gives as comparison the result of the
gradient-selected method requiring one-fourth of the experi-
mental time, which is identical except for some reduction in
signal-to-noise. This reduction is somewhat counterbalanced
by the slightly higher receiver gain possible with the gradient
method. Contrary to these measurements, a standard NOESY
sequence under the same conditions, using only a two-step
phase cycle for axial peak suppression, revealed the known
COSY-type signal breakthroughs. These artifacts may become
smaller toward longer mixing times; with the gradient method,
however, one can be sure that these unwanted signals will be
suppressed. Our new method will not be necessary for those

FIG. 2. (a) Expansion of the aliphatic region of a standard 2D NOESY measurement on 10% strychnine ( 1) in CDCl3 , using an eight-step phase
cycle, 2K data points in F2 , 256 data points in F1 , recorded with a spectral width of 10 ppm (Bruker AMX-500 NMR spectrometer). (b) Same conditions
as in (a), recorded with the gs-NOESY method of Fig. 1 with a pulsed field gradient of 250 ms length and strength of approximately 0.01 T/m.

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 COMMUNICATIONS
FIG. 2Continued
who must accumulate NOESY spectra with many scans, but
in summary we feel that the gs-NOESY method is a significant
step toward improving the throughput for NOESY spectra in
service laboratories for organic chemistry.
ACKNOWLEDGMENTS
This work has been supported by the Deutsche Forschungsgemeinschaft
and the Fonds der Chemischen Industrie.
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