(These instructions assume you are familiar with basic Macintosh procedures, if you are not, see a Macintosh manual or the tutorial which is on most Macintosh computers. Throughout the following, MB means mouse button.)
Open the standard H1 setup file (your lab folder should have a H1 setup file, if it does not, find one in the setup filesgn folder). Collect and process a 1D spectrum (for directions see Acquisition and Processing of 1D H-1 Spectra). Zoom on a peak which you wish to irradiate. Move the cursor to the center of the peak, hold down the shift key and click the MB. The decoupler frequency will be set to the position of the cursor that appears (ie. in the center of the peak). Move the cursor to the Dashboard Button at the top left of the MacNMR window and click the MB to open the Dashboard window. Record the F2 offset value (this is the distance from the center of the spectrum to the decoupler frequency). Zoom on any other peaks which you wish to irradiate during the NOE experiment; determine and record their F2 offsets as above. Also zoom on one or two baseline areas which contain no peak intensity and record a F2 offset from this(these) region(s). Look at the Dashboard window and record the RCVR gain value. Move the cursor to File at the top of the screen, hold down the MB and drag to close.
Open the 1dnoesetup file located in the setup filesgn folder. Open the Dashboard window and set the RCVR gain to the value which you recorded above. Open the Console window and then open the AIR window. Set the value next to Spinner to 0 by double clicking the MB on the value and then entering 0. The rate value should go to 0. Open the LOCK window and increase the Lock RX value so that the lock level is between 80 and 100. Set the F2 offset to the value recorded above for the broadest peak which you wish to irradiate. Save the 1dnoesetup file (make sure to use the same file name - 1dnoesetup and write over the old file). Execute the Zero and Go command to collect 4 scans, process the spectrum and look at the saturated peak. One would like the saturated peak intensity to be reduced 75% to 95% compared to the peak amplitude in the absence of saturation. If the amount of saturation is unsatisfactory, open the Console window and then open the FREQ window. Find the Decoupler Gain value in the DECOUPLER (F2) section of the Console window. Increase the value if you need more complete saturation, decrease the value if you need less saturation. Broad peaks require larger values than narrow peaks, a value between 23 and 35 should be appropriate. Exit the Console window.
Move the cursor to the Script Icon at the top of the screen (it is to the right of the Windows menu item at the top of the screen). Hold the MB down and drag to Launch Script Editor, release the MB. Move the cursor to File, hold down the MB and drag to Open Script, release the MB. When the dialog box appears, open the scripts folder in MacNMR-GN and then open 1D NOE Collection.
The script may need to be edited in four places. Make sure that line 3 is the following:
set my_file to "GN-300 NMR Mac:MacNMRGN:setup filesgn:1dnoesetup"
Edit line 4 to be the following:
set my_target_file to "GN-300 NMR Mac:MacNMRGN:Your lab:yourfile"
where Your lab is your lab folder name (ie. Harman labgn) and yourfile is the name of the file into which the data will be saved.
Move to the line that begins with:
set irradiation_offset_list to {
Delete the values in this list and enter the F2 offset values that you recorded above.
Move to the line that begins with:
Set NMR Parameter name "Scans 2D" value
and make sure the numerical value following the word value is set to the number of F2 offset values which you entered for the irradiation_offset_list.
Move the cursor to Check Syntax and click the MB. Close all windows that are open except the two script windows. Quit all applications (including MacNMR) except the Script Editor. Make sure the 1D NOE collection window is active and move the cursor to File, hold down the MB and drag to Save, release the MB. Move the cursor to Run in the 1D NOE collection window and click the MB. If a dialog box appears requesting you to select a file, move the cursor to the Cancel box in the dialog box and click the MB. The MacNMR window that appears on the screen must remain the top (active) window in order for script execution to occur. If you use the standard 1dnoesetup file and the standard 1D NOE collection script you can use the following formula to determine the approximate length of your experiment:
(20 sec.)x(value of Scans 2D)x16 = expt. time
After acquisition is complete, close any MacNMR files that are open and close the Script Editor. Open the NOE file that you collected, your file contains multiple spectra. Move the cursor near the slider and arrows located at the bottom left area of the MacNMR window just above Record. You can choose different spectra by putting the cursor on one of the arrows and clicking the MB or putting the cursor on the slider, holding down the MB and dragging.
Select a FID collected when the decoupler was set to a baseline position (this will be refered to as the control spectrum). Process this FID and adjust the phase. Move the cursor to Commands at the top of the screen, hold down the option key, hold down the MB and drag to nDFT, then release the MB. The Multi-dimensional Filter and Transform Settings window will appear. Move the cursor to the Load button at the bottom of the window and click the MB. Go to the setup filesgn folder and open the template file named 1dnoeprocess. Move the cursor to the box to the left of Revert to saved and click the MB (make sure an X appears in the box). Move the cursor to the Apply settings button and click the MB. Adjust the phase of the control spectrum and then repeat the nDFT command. All of the FID's in the file should now be processed and the phase should be properly adjusted. Save the file to a new name.
The NOE difference spectra can be obtained by using the Add/Subtract routine and subtracting the control spectrum from the spectra obtained with peak saturation.The Add/Subtract routine can be found under the Option menu item. See Chapter 5 of the MacNMR manual for Add/Subtract instructions. The difference spectra may be saved and the Peak Pick routine (found under the Option menu item, see MacNMR manual, Chapter 5, or Acquisition and Processing of 1D H-1 Spectra) may be used to obtain peak amplitudes. The NOE value is equal to the difference spectrum peak amplitude divided by the corresponding control spectrum amplitude. NOE's are usually expressed as decimal numbers or percents (ie. 0.010 = 10%).