NOESY

Set up appropriate 1H parameters. Normally should acquire 1D 1H spectrum first.

pw = 1H pulse, should be 90º pulse

tpwr = high power for 1H; should be set according to pw, or is correlated to pw.

nt = number of scans, should be multiple of 16, but 8 is acceptable

ni = number of t1 increments, points in t1 dimension, at least 200, 256 is
preferable

np = number of points in t2 dimension, must be even number, 2048 is standard

d1 = relaxation delay; normally should be longer than in gCOSY to decrease t1 noise, and make ROEs between different types of protons more equivalent, so 2.0-2.5 seconds is normal, but aromatic (or vinyl or methyl) protons usually need 3.0-3.5 seconds for better spectra

mixN = mixing time; the larger the mixing time the more NOEs will be observed, but the less difference in intensity by distance. In addition, the mixing time is correlated to the size of the molecule, small molecule, long mixing time; generally 200-300 ms (so mixN = .2 or mixN = .3) for MW ~400-2000 for longer mixing time 100-150 ms for short mixing time; 300-500 ms for long mixing time for MW < ~400


Steps:

1) Acquire 1 scan of 1H 1D experiment. Set cursors ~0.5 ppm beyond last proton resonance on both sides of spectrum, type command:
movesw

2) Re-acquire 1H 1D experiment (this is only required for 1H 1D on side of 2D spectrum)

3) Move parameters to another experiment. To move parameters from experiment 1 to experiment 3 type command:
mp(1,3)

4) Change to experiment 3, type command:
jexp3

5) Set NOESY parameters with setup macro, type command:
NOESY

6) Spin will likely turn off, if it does not, open Acqi window and manually turn it off. Lock Level should not drop more than ~5 %-10 %. If it does, you should reshim non-spin shims- X1, Y1, XZ, and YZ, and on higher field (500/600) X2Y2 and XY

7) Set the number of scans (nt) to appropriate value, generally 8 (it is somewhat preferable for nt to be a multiple of 16 but nt = 8 is acceptable; nt might need to be increased for sufficient signal-to-noise), type:
nt=8

8) Set the number of points in t1 dimension (second dimension) (ni), generally 200-400 are suggested, the greater the points, the better the resolution, and the less t1 noise observed, type:
ni=256

9) Set d1 time, normally should be longer than in gCOSY to decrease t1 noise, and make NOEs between different types of protons more equivalent, so 2.0-2.5 seconds is normal, but aromatic (or vinyl or methyl) protons usually need 3.0-3.5 seconds for better spectra

10) Set the mixing time (parameter = mixN); the larger the mixing time the more NOEs will be observed, but the less difference in intensity by distance. In addition, the mixing time is correlated to the size of the molecule, small
molecule, long mixing time; generally 200-300 ms (so mixN = .2 or mixN = .3) for MW ~400-2000 for longer mixing time 100-150 ms for short mixing time; 300-500 ms for long mixing time for MW < ~400

11) Start acquisition, type command:
go

12) To process data with VNMR type: processnoesy

13) If spectrum has yellow streaks and purple streaks horizontally across the spectrum, then the spectrum should be phased; set cursor on peak, type:
ds
then manually phase, be careful to only click once on spectrum during phasing, only rp (or rp1 phase correct should be changed)
redraw 2D type:
dpn10
If spectrum still has streaks in horizontal direction more phasing is required including lp (or lp1) phase correct. If spectrum has yellow/purple streaks in vertical direction, then the other dimension needs to be phased, so switch the axes, type:
trace = 'f1' (or 'f2', depending upon which axis is currently the x-axis, then type:
dpn10

14) Spectrum should be appropriately referenced already, but you should confirm this; if necessary re-reference by putting cursor on appropriate diagonal peak and type (assuming CDCl3):
rl(7.26p)
rl1(7.26p)
dp10

15) Adjust vertical scale with vs +20% and vs -20% menu buttons or with middle mouse button or manually changing the parameter, vs2d, so vs2d = 100 (the lower the number, the less noise displayed), then redraw 2D with dp10 command

16) or Print with full rectangle, type:
full
dp10
pcon(10,1.2) page

17) Save data, type:
svf('filename')