700 MHz NMR checklist
Check in for the NMR 700mHz
Make sure the system is accessible, e.g. not being used or under maintenance
Check your belongings on your body and remove anything with ferromagnetic properties such as electronic devices and keep them away from the probe
Go to the Sample Jet and change the operating mode to 5mm shuttle, then load the rack of NMR tubes.
Login to the computer
Start topspin software
Load sample by typing in “sx 101” (or if in rack 2: “sx 201”, etc)
Click on the blank document icon
- Name it: initials-title-date
- Experiment: zgesgp (wipes out the water peak)
- Set solvent
- Check the “getprosol” box
Lock the sample by typing “lock d2o”
- The signal should be at around 75% after you lock it. If it is too low, go to BSMS, click “power” and use the scroll on the mouse to get it to around 75%, then click “standby” on BSMS to save the change.
Shim the sample automatically by typing “topshim”
- If the signal is not at 75%, adjust it in the same way as before. Also you can adjust the shimming manually by going to the BSMS and clicking on “z1”, “z2”, “x”, “y”, “xy”, etc. dimensions and using the scroll on the mouse. Don’t forget to click “standby” after any changes.
Autotune the sample by typing “atma”. To manually autotune, type “atmm”.
Next you need to find the 90° pulse.
- Type P1 and enter “8.0” in the pop-up window
- Type “pulprog” and choose “zg”
- Type “ns 1” (number of scans=1), then type “ds 1” (dummy scans), then type “rg 1” (receiver gain).
- Run the sample by typing “zg”
- To transform FID into spectrum, type “efp”
- Type “apk” to autophase
- Not type “p1”, hit enter and try different numbers (usually between 30-44), type “zg” and “efp” in order and see what number (p1) minimizes the peak the most.
- Once you have found the p1 number that minimizes the peak, divide that number by 4. That number is your 90° pulse.
- Type “edprosol” and set the p1 you found into both pulse widths and hit enter in each one
- Click the “copy to solvent” buton and select all relevant
- Click the “copy to probe” and select all relevant again. A popup window will ask you which cryoprobe you want to use. Choose the one that matches the cryoprobe that is in the top left corner.
- Save then “select all relevant”. Click on “yes” and “ok” whenever it asks you and make sure that you calculate all pulses when it asks. Then close out.
To run IconNMR, type “iconnmr” and choose the automation option and enter the password
Go to the holder that your sample is in. For example if your sample is in the 1st rack and is in the first slot in that rack, then go to A1-101.
Type in the name of your experiment, click on the “No.” space to activate that box, choose your solvent, and choose your experiment.
- Experiment for HSQC: C13HSQCS1SP2 (metab)
- Experiment for HMBC: C-13 HMBC
- Experiment for HSQC-TOCSY: SL-hsqcetf3gpml. Metab
Click on the paramters tab and choose “edit all acquisitions parameters”
- In the TD (data points) for the H dimension enter a number that is between 1024 and 2048 although it’s usually 2048. In the C dimension enter a number that is between 64 and 256 although it’s usually 64.
- In the NS (number of scans), type 32 although it can be 64 and even 128 depending on what you want.
- Type d1 and hit enter and input 1.5 for the relaxation time. *If you are doing the HSQC-TOCSY experiment then make sure you also change the d8 (delay time) to whatever number corresponds to the right hand panel
- Type in “rga” to check the automatic receiver gain
- to automation tab and make sure that the AUNM is set to au-zgonly.
- Then click on the button that says “return to iconNMR”
Copy to the samples as needed. (“edit as needed” )
Highlight all of the samples and click submit
Click “Start” and check the “lock/shim has already been completed box” and make sure that you are starting at your sample (for example: 101).