How to acquire a 2D spectrum on the DPX300:

1. Sample setup:
• Insert, lock and shim your sample as described in the 1D guide .
• Stop spinning by clicking on the [Spin ON/OFF] buttton in the BSMS window (so that the yellow light is off)
  


2. 1D reference spectrum
   
• Define a dataset with new and take a 1D proton spectrum.
• optional: take 1D ¹³C or DEPT spectrum.


3. Set up 2D spectrum:
1. Type new to define a new data set.
2. Type rpar experiment all to read the parameter file for your experiment.
   Click here for list of available 2D experiments:
   An updated listing can also be found next to the spectrometer or by typing rpar *.hf all .
3. In most cases, you are ready to run. Depending on your sample concentration or unusual chemical shifts you may have to adjust some parameters:
   
   • sw, 1 sw: spectral width in the direct and indirect dimension to include all peaks in your 1D spectra
   • o1p, o2p: frequency offset for proton and carbon (or other hetero nucleus).
   • ns: number of scans per increment. Increasing improves signal to noise. If your sample is very dilute (solvent signal higher than sample signal) you will have to increase ns.
   • 1 td: number of increments. Increase improves signal to noise and resolution in the indirect dimension.
   • NOESY only: d8 NOE mixing time, may depend on size of molecule
   • TOCSY only: D9 set to 10 ms for COSY peaks only, 60-80 ms for relay peaks
   • Use expt to calculate the time required for the experiment. Total time will depend on both ns and 1 td
   • all these parameters can also be accessed by typing eda
   • use ased to get a list of only the acquisition parameters used by the experimennt
4. Type rga to adjust the reveiver gain of your sample.You can check the receiver gain by typing rg.
5. Start the experiment by typing zg.
6. Multiple experiments can be set up for use with multizg: Define the next experiment by repeating new, rpar and rga again as (same sample) described above. Keep the experiment name, but increase the number by one. Repeat this until you have defined all experiments.
   Then return to your first data set by typing re expno, where expno stands for the experiment number of your first data set.
   Start acquisition by typing multizg and enter the number of experiments you have defined.
   


7. Other useful acquisition commands :
• halt halt the current acquisition prior to finishing all experiments and save the fid.
• acqu or a switch to the acquisition display window to display the fid.
• rser 1 read the first fid and switches to the 1D spectrum. [2D] gets you back to the 2D spectrum.
4. Processing, manipulation and plotting:

This can be done both directly at the spectrometer or the external data station

1. Type edc2. In the window, define your proton spectrum as 2^nd data set. If applicable, define the ¹³C spectrum as 3^rd data set.
2. getref will copy 2^nd and 3^rd data sets as projection and calibrate 2D to match.
3. Type xfb to process the two dimensional spectrum. This can already be done while the acquisition is still in progress to
4. Type abs2, then abs1 to perform a baseline correction in each dimension.
6. Adjust the threshold to see all signals by keeping the left mouse button pressed and moving the mouse up and down
7. Zoom region: Click on the spectrum with the left mouse button, then define the box with the middle mouse button. Click to zoom the region defined by the box.
8. [+/-] Switch between display of positive, negative and both
9. Limits Define the plot limits and contour levels. You will be asked to enter your limits in ppm and the number of contour levels to be drawn, default is your current display
10. setti set plot title. This command calls up a text editor
11. view gives a preview of the plot
12. plot send the plot to the printer.


5. Finishing:
• ej eject your sample, insert standard
• ij to lower down standard
• turn spinning on by clicking [Spin ON/OFF] in BSMS window
• type finish
• Finish your log book entry