SOP for Bruker NMR spectrometer

OBJECTIVE

This SOP describes the procedure for the operation of Bruker 400MHz NMR.

SCOPE

This SOP applies to the procedures to run NMR experiments on the Bruker 400 MHz NMR.

RESPONSIBILITIES

It is the responsibility of the QC Analyst to ensure adherence to this procedure.

It is the responsibility of the QC Manager to follow the instructions of this procedure.

DEFINITIONS

NMR: Nuclear Magnetic Resonance.

DEPT: Distortion less Enhancement by Polarization Transfer

COSY: Correlation Spectroscopy

NOESY: Nuclear Overhauser effect spectroscopy.

PRECAUTIONS

ALWAYS SET DEPTH with a depth gauge!! Failure to set depth will break your sample and/or the instrument.

PROCEDURE

Turning on the TOPSPIN software

• Switch on the PC
• Wait until the login screen appears
• Switch on the Console
• Wait at least 3 minutes
• Login as "NMR User" and the PW is "topspin".
• Start Topspin 3.0.
• Ignore all error messages.
• Type "redhead" in the command line (at the bottom of the screen)
• Ignore the error message.
• The Edhead window opens Click on "exit".
• Channel Routing window opens click on "close".
• Type "ii restarts" in the command line
• The BRUKER 400 MHz NMR is ready to use.

Running the samples

• Select Start

• Click on "Create data set"
• Following window opens

• Enter name and experiment number
• Keep the PROCNO " 1" only
• Select the solvent that you are using from the drop-down menu.
• Experiment: use current parameters.
• Title: Enter a title and press OK
• New window opens

ALSO READ: SOP for Calibration of Water Proof Hand Held TDS Meter

To Read parameters

• Select Proton for 1H NMR, C13CPD for 13C NMR,
• DEPT : First perform C13CPD than DEPT 135p OR first DEPT 90 than
• DEPT135p.
• 2D NMR: COSYGPSW for (H, H COSY), Select HSQCGP for (C H COSY)

1. Click "Read Pars"
2. "rpar" window opens
3. Select all and click OK

Data Acquiring

• Preparation of NMR sample: Prepare the sample in the required solvent and fill it about 6 cm (4 cm to 8 cm, 6 cm is ideal). Insert the sample tube into the spinner.
• Keep the spinner in the sample depth gauge and press the sample tube till it touches the white base. The bottom level should be set to 2 cm from the bottom (White base).

ALWAYS SET DEPTH with a depth gauge!! Failure to set depth will break your sample and/or the instrument.

• Select Turn on sample lift air (ej) from the Acquire main menu.
• Drop the sample tube on the red sample holder (Top of the instrument)
• Select Turn off sample lift air (ij)
• Wait till you hear a tick sound.

Lock

• Click on lock
• The solvents table will open
• Select the solvent and press OK

Tune

• Click Tune on yellow and select Tune/match prob head automatically.
• Wait till the message "atma finished"

Spin

• Spin is required for 1D, for 2D 3D no need to spin
• Turn sample rotation on
• Wait till the red arrow appears on the sample

Shim

• Click on the shim yellow button
• Wait till the top shim completely appears.

Prosol

• click on prosol

Gain

• Click on gain and wait till finish.
• Check the scan time if required change. For 1H NMR set scan in multiples of 4 usually 16 scans.

GO

• To start the scan click GO.
• Click on Acqu from the inner window to see the FID.

Data Process, Saving, and Printing

• Click on Process main menu and select Proc spectrum

• Click the Adjust phase. Using the left button, scroll and adjust the phase. Click save + return.
• Click integrate. Click to define the beginning and end of the integral; press the left button and drag to define. Right-click under integral to select. Adjust selected integral tails to be parallel with the baseline by scrolling bias and then slope.
• To calibrate, click under integral and select calibrate. Click save+ return.
• Click pick peaks. Make sure the defined region is selected. With the left button drag the box to include the top of all peaks to be picked. To add a peak, click and select peak with the cursor and left-click. Click save+ return
• To save: Select Publish from the main menu and save by selecting PDF.
• To print: Go to print on the left hand,
• Options: Select Print with layout-start plot editor
• Press OK.

To turn off the instrument

• First, eject the sample by clicking on the acquire the main menu and selecting Turn on sample lift air(ej).
• Remove the sample and inject again by selecting Turn off sample lift air (ij)
• Click on the left-corner icon (yellow).
• Press terminate TopSpin and press OK.

ABBREVIATION

QC: Quality Control

ANNEXURE

Nil

REVISION HISTORY

Nil

ALSO READ: SOP for Operation of Shimadzu Atomic Absorption Spectrophotometer