User guide for the NMR spectrometer AVA400Stud
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1 1 User guide for the NMR spectrometer AVA400Stud Dear User! You are using a modern NMR spectrometer, which has been installed in About spectra are measured here per year. Please respect the interests of other users and read the entire manual before using the spectrometer. Nevertheless, nobody is perfect and we are all learning from mistakes. Please report such cases immediately to the NMR department to avoid spectrometer downtime or even damage. Do not waste your time with trying to delete the traces of your activities. You will not be punished! On the contrary, your courage and responsibility can help tremendously in troubleshooting problems caused by your error. I. Safety precautions The superconducting magnet of NMR equipment produces strong magnetic and electromagnetic fields that can interfere with the function of cardiac pacemakers. Users of pacemakers and other implanted ferromagnetic medical devices are not allowed to enter the NMR room! Pregnant women and persons with non-ferromagnetic medical auxiliaries are required to contact the NMR department before using the spectrometer. The force that the magnetic field exerts on a ferromagnetic object (iron) may be very strong. Loose metal objects in the vicinity of the magnet can make sudden movements and be attracted to the magnet. This especially applies to very small ferromagnetic objects, which can be drawn into the magnet. They can damage the probe, are very difficult to remove, and will block the use of the spectrometer for a long time. The magnetic field can erase the magnetic media of USB-sticks, MP3-players, and mobile phones, disable credit and automated teller machine (ATM) cards, and damage analog watches. Please leave such items at the entrance of the room. The wall thickness of an NMR tube is about of 0.4 mm. Do not apply excessive force on when you close, open or insert the NMR tube into the spinner turbine. In rare cases, the superconducting magnet can spontaneously change from the superconducting to the normal state ("quench"). This transition causes an explosive vaporization of cryogenic substances from the magnet. In such case you are obligated to leave the room immediately and to inform the NMR department.
2 2 II. NMR sample preparation and injection 1. Samples for NMR analysis should be prepared in a defect-free 5 mm NMR tube. The tube cap must be on tightly to prevent evaporation of solvent while waiting in the queue. 2. The use of sealed or J. Young NMR tubes is prohibited. 3. Either 7 or 8 inch NMR tubes can be used and the tubes must have a quality according to the fieldstrength (e. e. 507-PP or higher). 4. Samples must be prepared using deuterated solvents. It is prohibited to use acids as the NMR solvent! 5. The optimal height of the liquid in the NMR tube is about 4.5 cm that corresponds to about 0.7 ml of the solvent. 6. The required concentration of an NMR sample depends on the experiment and the sensitivity of the observed nuclei. If possible, increase the concentration to improve the quality of the spectrum, especially for 13 C experiments. 7. The solution must be homogeneous. Precipitation and air bubbles impair the quality of NMR spectra and waste valuable measuring time. Solvent mixtures are avoided whenever possible. 8. NMR tubes must be clean outside. 9. Always wipe the NMR tube with Kleenex before inserting it into the spinner turbine. 10. Spinner turbines can be touched exclusively with clean, grease-free fingers at the upper edge (barcode). 11. The position of the NMR tube in the spinner turbine must be set using the red depth gauge. Slide carefully the NMR tube into the spinner turbine, plug the NMR tube along with the spinner turbine in the depth gauge and slide the tube gently to the bottom of the depth gauge. If you notice that the NMR-tube is loose inside the spinner then try another spinner or put your sample in another NMR-tube. Not correctly adjusted NMR tubes can lead to costly damage of the NMR spectrometer! 12. Place your NMR sample in the next free holder of the sample changer carousel. Please avoid the area of the light sensor to prevent the malfunction of automation. Use the holder number to register the sample in ICON-NMR. Caution: If the arm of the changer is moving or the carousel is rotating it is vorbidden to put a sample into the carousel or to remove a sample from it! Note the different holder numbers of the inner and outer chains of the carousel (see the schema on the sample changer). Experiments are started according to the required measuring time not the holder number (day experiments from 7 to 19 o clock and night experiments from 19 to 7 o clock).
3 3 13. Please always check the proper adjustment of your NMR tube by comparison with other NMR samples (visual control). 14. Each NMR sample must be accompanied with a perforated note of reasonable size providing the username, the sample composition, and the holder number. III. General rules Please respect the interests of your colleagues. You are not the only ones who are in a hurry. ICON-NMR shows the available measurement time for the daytime and nighttime experiments as well as the expected duration of experiments. On the weekdays (from Monday to Friday) the pending queue of nighttime experiments is terminated at 7:30 o clock. The uncompleted experiments have to be resubmitted for the next night! 1. Warning: In case ICON-NMR is failed to start three consecutive NMR samples (incorrect number holder, inhomogeneous solution, too little solvent), the automation will be stopped for safety reasons and further measurements will not be carried out. 2. Please collect the measured samples as soon as possible.
4 4 IV. Referencing of NMR spectra on AVA400Stud 1 H-Spectra: Use the solvent peak as the reference. 13 C-Spectra: Use the solvent peak as the reference. In D 2 O use the SR-value of -6, P- and 19 F-Spectra: The lock signal should be a reasonable size. The SR-value depends on the solvent: Solvent SR-Wert für 31 P SR-Wert für 19 F SR-Wert für 11 B Aceton-d 6-270,95-397,36-103,08 CDCl 3-110, , CD 2 Cl 2-99,48-37, ,21 CD 3 CN - 242,13-457,29-80,27 C 6 D 6-109, , ,92 D 2 O - 116, , ,79 DMSO-d 6-196,95-162,57-43,07 CD 3 OD* - 225,15* - 611,55* - 66,80* *the SR-value is valid only if the methyl resonance is used to lock the spectrometer!
5 5 V. Instructions for ICON-NMR 1. Use the "Change User" to select your work group. 2. Locate the next free place in the sample changer carousel. The colors indicate the following: Yellow: occupied, experiment done. Rot: occupied, experiment in progress. Pink: occupied, waiting to run. Green: free, experiment may be set up. Note: If all the 120 positions are occupied, do the following: a) Starting from the position 1 locate the first yellow position. X b) If the latter position in the carousel is still occupied by an NMR sample, proceed to the next position labeled in yellow in ICON-NMR. Thus, locate the first position which is labeled in yellow in ICON-NMR and is not occupied by an NMR sample in the carousel. c) Delete the corresponding record in ICON-NMR list with "DELETE". d) Place your sample in this position on the carousel. 3. To submit your experiment make a double click on "STATUS" and fill out the form that appears: Holder: The holder number. Type: Prescribed. Status: See item 2. Disk: Prescribed. Name: Generated automatically. Exp. No.: Generated automatically. Solvent: Define from the list. Experiment: Define from the list. 4. Check whether the holder number in ICON-NMR corresponds to the one in the carousel. Submit your experiment using the button "SUBMIT" at the middle of the left side of the screen.
6 VI. NMR experiments and the required concentrations of samples 6 Title of the automation: NS: Sweep width [ppm]: Minimal concentration [mol/l]: Approximate measurement time (without Shiming!): 1 H rau_sprotonlf_16 NS minutes rau_sprotonlf_64 NS minutes 13 C{ 1 H} rau_sc13cpd_256 NS minutes rau_sc13cpd_1k NS 1k minutes Dept135 rau_sdept135_64 NS minutes rau_sdept135_256 NS minutes Dept90 rau_sdept90_64 NS minutes rau_sdept90_256 NS minutes 31 P{ 1 H } rau_sp31cpd_64 NS minutes 31 P rau_sp31_64 NS minutes 19 F{ 1 H } rau_sf19cpd NS minute 19 F rau_sf19 NS minute HSQC rau_shsqcedetgpsw NS 2 Optimized in F2 in F1: 0 165ppm minutes HMBC rau_shmbcgpsw NS 2 Optimized in F2 in F1: 0 220ppm minutes Cosy rau_scosygpsw NS 1 Sweep optimized minutes Noesy rau_snoesygpsw NS 2, d8=1s Sweep optimized minutes Roesy rau_sroesysw NS 8, p15=0.2s Sweep optimized minutes Tocsy rau_stocsysw NS 8, d9=0.08s Sweep optimized minutes
7 7 VII. 2D-Spectra: Each 2D-experiment must be preceded by a 1 H experiment (rau_sprotonlf_16) based on the same set-up file. The later spectrum is required to define automatically the measuring range for the 2D spectrum. Upon the submission of a 2D-experiment ICON-NMR will generate automatically some parameters in the Setup Form. Please do not change these parameters to avoid a malfunction of automation. HSQC and HMBC: The measuring range of the direct dimension F2 (protons) is automatically optimized using the 1 H NMR spectrum (rau_sprotonlf_16). The measuring range of the indirect dimension F1 ( 13 C) is prescribed. COSY, NOESY, ROESY, and TOCSY: The measuring range of both the direct and indirect dimensions is automatically optimized using the 1 H NMR spectrum (rau_sprotonlf_16). Par. : Parameters. If necessary define d8 and d9 for NOESY- and TOCSY experiments. (Note: the values are in seconds!) Orig./Title: Enter your Status (Student, Bachelor, Master), your name, the substance, and the concentration: e.g.: Bachelor, Max Mustermann, TB45 (10mg/0.8ml CDCl3). Pri.: Display of Day- or Nighttime Experiment, otherwise disabled. Time: Display of the measuring time. (Note: add up to 5 minutes for Lock, Shim and the thermostating of the sample. User: Display of your username.
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