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Category
- Spectroscopy and Spectrometry » NMR Spectroscopy
Booking Details
Facility Management Team and Location
ishita@chem.iitb.ac.in
suvarn@chem.iitb.ac.in
shutoshk@iitb.ac.in
chandra.volla@chem.iitb.ac.in,
eswar@chem.iitb.ac.in
arnabdutta@chem.iitb.ac.in
Facility Features, Working Principle and Specifications
Facility Description
This facility enables the recording and analysis of a number of conventional 1D and 2D NMR experiments for liquid organic/inorganic small molecule samples as well as 3D measurements for protonated and isotopically labelled biomolecules, below ~15 kilodaltons. These experiments are vital for identification, assignment and structural/dynamic characterization of a wide range of molecules.
NMR spectroscopy is a high resolution analytical method for structure, conformational analysis and dynamics of molecules. NMR-active nuclear spins are excited by the application of radio-frequency pulses of matching frequency. The magnetic field generated by these spins is detected by the same RF coil which is used to excite them, giving rise to the NMR signal (FID). Signal processing results in a frequency domain spectrum which is then analyzed to reveal information about the environment around the spins.
Probe Specifications:
1.Bruker 5 mm SmartProbeTM (standard room-temperature BBO probe) for multinuclear measurements of small molecules. The probe can be tuned to observe 19F or any nucleus in the range from 31P- 199Hg and 17O-109Ag excluding 171Yb-9Be. Simultaneous pulsing on 1H and 19F nuclei is possible, enabling the recording of 19F spectra with 1H decoupling. The probe is equipped with a z-gradient (5 G/A*cm), 2H locking and is capable of carrying out solvent suppression using pre-saturation or pulsed-field gradients.
2. 5mm Triple resonance (TXI) high-resolution room temperature-probe (with actively-shielded 5 G/A*cm z-gradient and 2H lock). The probe can be used for structural and dynamic analysis of doubly (13C/15N) or singly (15N) labelled biomolecules, using 1H detected measurements and 13C/15N decoupling.
Instructions for Registration, Sample Preparation, User Instructions and Precautionary Measures
Internal Users
Users within IIT Bombay can apply using https://rnd.iitb.ac.in/nmr_600_mhz_spectrometer
Please click on this link to see the flow chart of the full procedure: @Link to Flow Chart
The online registration form should be completely filled out by the applicant. For any query contact the facility manager.
After getting approval from the student’s PI along with the committing charges, an appointment will be given as per queue, and will be informed by email/phone.
At least 24 hours prior information should be given to the NMR lab if users want to cancel/postpone the slot.
External Users:
1. External users can come personally or send the filled in registration form along with the Demand draft and a letter from the Guide / Head on the Institute Original Letter Head stating that the analysis is for research purpose, to qualify for academic concession. The letter should be addressed to “Prof. Ishita Sengupta, The Facility-in-charge, 600 MHz NMR Facility, Department of Chemistry, IIT Bombay, Powai, Mumbai – 4000 76.”
2. Samples can be brought along on the date of your appointment for your sample analysis or can also be sent to the following address: “Prof. Ishita Sengupta, The Facility-in-charge, 600 MHz NMR Facility, Room no. 117, Ground Floor, CRNTS, SAIF Building, IIT Bombay, Powai, Mumbai – 400076.”
3.Users can use one registration form for a maximum of three samples.
Inorganic/organic small molecule samples which are highly soluble (minimum concentration 1 mM- 2mM) in typical NMR solvents (deuterated methanol, deuterated DMSO, deuterated acetone, deuterated chloroform or D2O etc.) can be analyzed. The molecular weight and solubility must be clearly stated when sending in samples.
Instructions for preparing NMR samples
- Use the correct quantity
Use the appropriate amount of material for the type of spectrum you're taking.
- Remove solids
Remove any solid particles from the sample. You can filter the sample with cotton wool or membrane filters.
- Use deuterated solvents
Dissolve the sample in a deuterated solvent, such as CDCl3, DMSO-d6, CD3OD, Acetone-d6 etc.
- Prepare the sample in a secondary vial
Dissolve the solid sample in a small vial and then transfer it to the NMR tube.
- Treat the sample
Treat the sample with heat or vortexing to ensure complete dissolution.
- Use clean tubes and caps
Use clean NMR tubes and caps. Avoid contact between the sample and the cap, or use new caps.
- Label the sample
Label the sample tube with the sample name and your group's short-form. You can also attach a flag with the sample name to the sample.
- Use the right tube
Use an NMR tube with a diameter of 5 mm and a minimum length of 17 cm. The tube should not have any glass defects.
- Fill to the correct height
The minimum filling height for an NMR tube is 5 cm (0.6 ml).
Isotopically labelled samples (15N labelled or 13C/15N labelled samples) can be sent in lyophilized form or on ice in the relevant buffer. If sent in lyophilized form, please include the NMR sample buffer. Sample concentrations should be 500 μM or more for 2D experiments and 1 mM or more for 3D measurements. Currently, long experiments are possible only at room temperature, depending upon the stability and solubility of the biomolecular samples.
1. External users are requested to mention in your request letter that “We agree to acknowledge the 600 MHz NMR Central Facility of IIT Bombay in our Publications / Reports / Thesis”. The information on such acknowledgements with appropriate reference should be communicated to the 600 MHz NMR, Central Facility lab via email nmr600ioe@iitb.ac.in. Kindly send the complete publication reference (Journal name / volume no. / page no. / date of issue of the publication, etc.
2. If you are sending samples by post please mention sample details:
a. Concentration (solution) or amount in mg (powder to be dissolved)
b. Sample name
c. deuterated (for locking) and regular solvent in which experiment should be carried out
d. Experiments to be carried out
e. If possible, experimental parameters can also be mentioned (particular scalar coupling constants etc.)
f. Temperature at which the experiment will be carried out
3. Once we receive the requisition letter and advance payment, your appointment will be scheduled as per the queue.
4. The users will be informed about their date and time of slot by e-mail.
5. NMR tube(s) and deuterated and regular solvent(s) should be included or if paid for, should be mentioned
6. We prefer that you or your representative, who knows/understands the sample/material should be present on the day of appointment.
7. All NMR users will be expected to bring/send their own NMR tubes and solvents (deuterated). The facility will charge extra for solvents and NMR tubes.
8. Rates are exclusive of GST. 9% CGST and 9% SGST for users within Maharashtra, and 18% IGST for users outside Maharashtra will be added to the total user charges.
9. 12 hours slots will be overnight slots exclusively. 24hour slots will be on weekends preferably. These will be assigned based on availability and in order of online requests. 12 hour and 24 hour slots will be used to run experiments on a single sample only (multiple measurements on the same sample possible). The longest booking slot is 24 hours at a stretch.
10. Booking should be done via the online portal only. No personal requests will be entertained. Slots will be assigned based on availability and in order of online bookings.
11. For Variable temperature measurements, please contact and confirm before hand for slots before booking – Liquid nitrogen charges extra. Currently, for low temperature measurements, a liquid nitrogen exchanger is available.
Charges for Analytical Services in Different Categories
Applications
NMR experiments:
Small molecules:
- 1D 1H with and without solvent suppression
- 1D NOESY, 1D DOSY, 1D CPMG (for T2 measurement), 1D Inversion Recovery (For T1 measurement)
- 1D 13C, 1D 19F, 1D 31P (coupled and decoupled versions), DEPT90, DEPT135
- 2D - NOESY, TOCSY, COSY, DQF-COSY, HETCOR, HSQC, HMBC
- 1D experiments on other X-nuclei (direct detection) - on request
Biomolecules:
- Standard 15N-1H HSQC, 13C-1H HSQC experiments
- 3D experiments for protein backbone and sidechain assignments (HNCO, HNCA, HN(CO)CA, HN(CA)CO, HNCACB, CBCACONH, CC(CO)NH, TOCSY-HSQC etc.)
- 3D 15N/13C edited NOESY-HSQC experiments for distance restraints
- Dynamics: Backbone (15N) R1, R2, hetNOE experiments
Variable temperature measurements:
For low temperature measurements, a liquid nitrogen exchanger is currently available. Liquid nitrogen charges will be extra. The user needs to inform at least 10 days in advance if they plan to carry out VT experiments, the details of which have to be discussed before proceeding, depending upon the current status.
Sample Details
Non-toxic solid and liquid sample
No