A. BASIC OPERATIONS

A .I Locking and Shimming the Sample

  1. Insert the sample into the magnet, spin the sample at 20 ±2 Hz and keep the temperature constant (300 K).
  2. Read the shim matrix file for the solvent you use:

    3.     RSH S05CDCL3 « (i.e. for chloroform, see table 1 for other solvents)
  3. Press "LOCK POWER" and turn the wheel to set the correct value for your solvent. Then press "FIELD" and turn the wheel to center the lock signal on the graphics display. (Start with the default settings from table 2.)
  4. Press the "AUTO LOCK" button and wait until its diode and the "LOCK GAIN" diode stop flashing.
  5. Adjust "LOCK PHASE" to maximize the position of the lock signal on the screen. If the lock signal leaves the screen, then decrease "LOCK GAIN" to let it reappear.
  6. Manual shimming:

    7.     Adjust the "Z" and "Z2" shims for a maximum position of the lock signal.
  7. Press the "STDBY" button to deactivate the wheel.
  8. For overnight experiments only:

    9.     Press "AUTO SHIM", "Z", "Z2", "AUTO SHIM".
Notes:

A.II Routine 1H and 13C Experiments:
  1. Read the parameters for the experiment

    2. RJ JH5CDCL3«         (for a 1H experiment with a chloroform-d solution.
    PJ « «                            See table 1 for other solvents.)
    II «
  2. Type: ZG « (wait until NS scans have been acquired)

    3. (If the FID is clipped horizontally, decrease RG and repeat ZG«)
  3. Save the FID: WR filename.extn «
  4. Manipulate the FID:         13C: Set LB = 2, Type EM«

    5.                                        (1H: Set LB = -1, GB = .25, type GM«
                                             You may skip this step for 1H, but not for 13C)
  5. To zero-fill the 1H FID, type: SI « 32 «
  6. Type: FT «

    7. Type: APK «
    (If the spectrum has a rolling baseline, decrease RG and repeat from step 2.)
  7. Type: ABS INT1.001 «
  8. Enter the Manual Processing Mode, if necessary.(EP «)
  9. Select what to plot:

    10. Plot the spectrum only:     PX «
    Plot the integral only:        PXI INT1.001 «
    Plot both:               ;          PXB INT1.001 «
  10. Eject the paper:     NP «
Notes:

A.III    Steps for Manual Processing of NMR spectra

Type "EP «" to enter the expansion and processing mode.

Phase correction (in the EP mode):

  1. Move the curser to the first strong signal in the spectrum. Press "P".
  2. Press the vertical display (+) button, until the noise covers half a box of the grid.
  3. Turn knob C to properly adjust the phase of the signal at the cursor position.

    4. (If you reach the end of the range for knob C, then press Ctrl-C to reverse it.)
  4. Move the spectrum to the last strong signal in the spectrum (knob A).
  5. Turn knob D to properly adjust the phase of this signal. Do not change C anymore!

    6. (If you reach the end of the range for knob D, then press Ctrl-D to reverse it.)
  6. Press "M" to store the phase correction into the memory.
Calibration of the ppm scale(still in the EP mode):
  1. Find the solvent signal or the TMS signal, if present.
  2. If all the values on the screen are displayed in Hertz, press ":".
  3. Place the cursor exactly on top of the signal and press "G".
  4. Type the correct chemical shift value for this signal (i.e. "7.25p" for chloroform-d).
Integration of the spectrum (still in EP mode):
  1. Press "I" to enter the integration routine. Begin with the signal at LOWEST field.
  2. Place the cursor to the left of the signal.
  3. Press "Z" to mark the beginning of the integral.
  4. Move the cursor to the right of the signal and press "Z" to mark the end of the integral.
  5. Repeat steps 2 to 4 for each signal of interest.
  6. You can normalize the integrals, if you move the cursor onto the last data point of an integral, type "A" and enter a number to define the area of this integral.
  7. After defining the last integral press "E".
  8. Press "«" to accept the displayed filename or change it. This is the file which you will use to print out the integrals with PXB or PXI. Remember the name!
Defining the plotting parameters (still in EP mode):
  1. To define the plotting range press "F" and enter low and high field limits in ppm.
  2. To define the minimum intensity for 'peak picking' place the cursor on top of the smallest signal to be picked. Press "M" and re-enter the displayed value.

    3. Terminate with: "P" to pick positive peaks only (for normal 1H or 13C)
                       or "N" to pick negative peaks only
                       or "«" to pick positive AND negative peaks (i.e. for DEPT)
  3. Choose:
    1. To define the height for one particular peak place the cursor on top of it and press "CY" without return. Enter the height of the signal and then the length of the plot in centimeters.
    2. To define the height for the tallest peak in the region visible on the screen press "Y" and enter the corresponding values.
  4. The rest of the parameters have to be entered outside of EP. Press "«" to exit.
  5. Set MAXX = 25, X0 = 0, Y0 = 0, CX = 25. Set MAXY = 14 for 1H and = 17 for 13C. If you have NOT defined the height of the signals inside EP, set CY = 12.
  6. Enter "PEN «" to define the colors for spectra, ppm-scale, integral, etc. Use numbers 1 - 7 according to the position of the pens in the plotter's caroussel.
  7. Enter "DPO «" to define the 'digital plotter options'. Answer all the questions so that you will receive the desired output of your spectra.

    8. Notes:     -if you want to print the integration, the 'offset' has to be at least 2.5 cm.
                   -with CX >20 the parameters have to be printed in the upper left corner.

A. IV    Automated Procedures

a) Locking and Shimming:
 

  1. Adjust the "Field":    4310 for DMSO         4400 for Methanol

    2.                                 4310 for Acetone        4680 for Chloroform
                                    4490 for D2O             4680 for Benzene
                                    4800 for Pyridine
  2. Type: RSH S05solvent « (use the proper filename for your solvent)
  3. Type: LOCK «
  4. Select the shimming method:

    5.     TU4  «      for quick shimming
        SH4  «      for accurate shimming
  5. Wait until the normal spectrum returns on the screen and the LCD shows the message "CONVERGED" or "READY TO MEASURE".
b) Automated Measurements:
  1. Read the job parameter for your solvent:

    2. i.e. RJ JH5CDCL3 «
  2. Type: PJ « «
  3. Type: II «
  4. Type: SINO « 4 « Y «
  5. Type: AU ROUTINE.AU «

    6. The program requests a filename to save the FID. DO NOT enter any extension!
    The program will stop, when a sufficient signal-to-noise ratio has been achieved,
    but may be interrupted any time by pressing CTRL-H.
    Make sure that the plotter has loaded paper and that there are still some pages left in the tray. Use a black pen in position 1 and a colored pen in position 2.
  6. Choose:

    7.     EXE PLOTH.EXE «    for a full 1H spectrum with integration.
        EXE PLOTC.EXE «    for a full 13C spectrum.
        EXE PLOTHEXP.EXE «     for expansions of a 1H spectrum.
        EXE PLOTCEXP.EXE «     for expansions of a 13C spectrum.
    Each program will ask for the filename of the FID.
    The file must have the extension .001, but do not enter the extension.

A.V Switching between NORMAL and REVERSE Operation

Please ask the NMR specialist before you do this!

- NORMAL to REVERSE

  1. Type "PO « PR « 0"
  2. Open the metal door at the right side of the AM-360 console.

    3. Pull out the "NORMAL MODE / INVERSE MODE" plug (upper left corner).
    Turn the plug upside down and plug it back in.
    (The upper label shows the current mode.)
  3. At the preamplifier housing (box on the floor next to the magnet) switch the following cables:

    4. a) Connect the cable labelled "Preamp.2" with the cable "F1IN" using a BNC connector.
    b) Plug the BNC cable "F2IN" into the plug "Transm. F1" at the side of the box.
  4. Type "PR « H1"
The NORMAL 1H and 13C experiments are NOT possible in reverse mode!
You need special microprograms to detect 1H.

- REVERSE to NORMAL

  1. Type "PO « PR « 0"
  2. Open the metal door at the right side of the AM-360 console.

    3. Turn the plug "NORMAL / REVERSE", so that the label "NORMAL" is on top.
  3. Plug the cable "F1IN" into the "TRANSM. F1" port on the preamplifier housing box.
  4. Plug the cable "F2IN" into the "DECOUPLER IN" port.
  5. Plug the cable "Preamp.2" into the "PREAMP.2-SELECTIVE" port.
  6. Type "PR « 11"

A.VI Leaving the Spectrometer:
 
  1. Put the D2O sample into the magnet.
  2. Type: EXE FINISH «
  3. Verify the settings of the Temperature unit:

    4.         Temperature: 300 K
            Heater Power Limiter: 6 to 7
  4. Please remove used tissues, used plotter paper and plotter pens from the console.
  5. Remove your samples as soon as possible from the spectrometer. Unclaimed sample tubes will be discarded after one month.
  6. Write your experiments or other activity at the spectrometer, their duration and the status of the spectrometer into the log book.
  7. Please report any kind of problems and irregularities.
  8. The door to G-14 has to be locked all the time, not just during the night!
  9. The easiest way to do this is to lock it immideately after you enter the lab, but also check it when you leave the lab, please.
 

(C)1998    Jürgen Schulte