© Bruker BioSpin 2008; File: braumann_jena_080828.ppt; Version: 2008-08-28; By: Dr. Ulrich Braumann LC-(SPE)-(Cryo)NMR(/MS) as Routine Analysis Technique:

© Bruker BioSpin 2008; File: braumann_jena_080828.ppt; Version: 2008-08-28; By: Dr. Ulrich Braumann

LC-(SPE)-(Cryo)NMR(/MS) as Routine Analysis Technique:Instrumentation and Software

Dr. Ulrich BraumannBruker Biospin, Rheinstetten

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Agenda

• Startup SampleTrack

• Startup Preparation Program PrepGilsonST

• Startup NMR Program TopSpin and ICONNMR

• Setup Orders


Startup instruments and prepare the System


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Prepare SampleTrack PC

• Login to the SampleTrack PC, typically with

- User: samtrack

- passwd: bsts

• Check that the following programsare running

- SampleTrack Webservice

- SampleTrack Communicator

- SampleTrack Client

• If not, start the programs

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• In the SampleTrack Client check go to the page instrument status

• Typically all instruments will show“not responding” as the programs onthe other computers are not started

• Follow the instructions on the nextpages to startup the instrumentprograms and in parallel observe inthis window that the connectionsare established.

• The instrument programs whichcommunicate with SampleTrack are

- Compass Sample Track communication

- ICONNMR

- PrepGilsonST

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• Clean up Sample orders

- Attention: During the next steps the automation programs are started.

- If they detected the barcode of samples which have open orders in SampleTrack, the preparation or NMR/MS measurement may start immediately.

• Therefore it is recommended to search for open orders from previousruns and –if required-delete them.

- Go to the pageSample Orders

- Search forNot yet finished Samples

- Check if this orders shallbe really executed. If notdelete them.

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Prepare PrepGilsonST

• Start PrepGilsonST, and select the“Profiler” application.

- Gilson remain disconnected– This is normal!

- A white bar must indicate aof SampleTrack and PrepGilsonST

- In SampleTrack Client must change from “not responding”to “preparation loop not started”

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Start PrepGilsonST

• Do Preparation/Start preparation Automation

- The program must connect to the liquid handler.

- The LiquidHandler must start reading barcodes.

- In the SampleTrack Client the status must turn to green

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Prepare ICONNMR & SampleJet

• On the TouchScreen of the SampleJet check if the correct mode is selected

- Do not use “manual mode” – switch to the correct shuttle diameter.

- Ensure that you have selected the correct diameter.

- Remove any sample from the magnet and ensure that the shuttle with the correct diameter is in the magnet.

• Open the ICONMR- Configuration

- Load the configuration for the SampleJet automation

- Check settings for locking, shimming, ATMA.

- If shimfile-loading is activated, check that the define shimfile is up-to-date.

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Start ICONNMR

• Start ICONNMR automation, do not define any experiments

• Start the automation procedure

- Select “SampleTrack(SampleJet).

- In SampleTrack the status must turngreen “online waiting for orders”.

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Prepare HyStar & MS Control

• Identify the standard method path ofHyStar (“D:\data” or “D:\methods”).

• From this path open the SampleTable“openaccess.xml” – or create it.

• The SampleTable should have one lineand the method should be thesame as used during the analysis.

• Enter the acquisition window ofHyStar with this line.

- The Acquisition window must openand HyStar must show green status.

- The MS control software must correctlyopen and show green status.

- If you cannot open HyStar/MS control withthe method selected for the measurements,STC will not be able either!

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Prepare HyStar & MS Control

• Start the program SampleTrack Communication.

- Start in online mode = yes

• HyStar will vanish in the background.

• The program must start and show status ready for all parts.

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Finish

• All instruments are now ready to run experiments.

• If the any experiment start immediately

- you have samples in the Liquid handler, NMR sample changer or LC-MS changer which still have open order in SampleTrack.

- Go to page Prepare SampleTrack PC and check if the listed orders are still wanted. Either delete the orders or remove the rack with the corresponding samples from the instruments.


Process Samples


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Setup Orders - manual

• Have the barcoded rack/wellplate with the samples ready.

• In SampleTrack do File/New

- Enter the ID of the tay (upper text like “RCK000183”)

- Enter the geometry identification of the tray.

- Select the sample position in the tray from the displayed layout.

• Select the “supermethod” that defines the complete process from the preparation to the measurment in the NMR and LC/MS

- If required customize the supermethod, for example to disable certain NMR experiments or disable the NMR or MS measurement part completely.

- Note: Disabeling the prepartion is not possible, as this also includes the assigment of the ID of the original sample to the position of the SampleJet tube/LC-MS well.

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Sample Preparation

• Open the door of the hood

- Place the tray with the samples into the liquid handler.

- Ensure that an unused rack with SampleJet tubes and a unused wellplate for MS aliquot is available.

(Note: If a number of samples already were prepared from the source rack, the original destination rack(s) must be available. I.e. you may have to return them from the SampleJet or LC-MS autosampler)

• Close the door of the hood

- The liquid handler must start scanning the barcodes to identify the new samples.

- The liquid handler will look for available destination racks.

- The preparation will start automatically.

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Sample Preparation

• The destination for a sample preparation will be automatically determined by sample track.

• Samples will be prepared symmetrically from source to destination.Sample #1A1, #2A2, #13 B1 etc (i.e. not submitted sample position will remain empty in the destination).

• The destination rack will remain reserved for the source rack.

• As consequence.

- You can remove a not completely filled destination rack from the liquid handler to start the NMR or LC-MS measurements.

- The preparation will continue into another free rack (if a suitable rack is found).

- However the empty spaces in both destinations racks are not available for other source racks!

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NMR Measurement

• Remove samples from the liquid hanlder

- Open the door of the hood.

- Remove the a rack with the SampleJet tubes.

- Close the door of the hood.

Note: If the preparation of one rack is not completely finished, the preparation will pause until the original SampleJet rack is back in the liquid handler!

• Insert the tray into the SampleJet

- Ensure that current shuttle diameter matches the samples tube diameter.

- At the SampleJet touchscreen select “Load rack”.

- Select the position and place the rack into the caroussel.

- Confirm with done.

- The SampleJet starts automatically to read the barcode of the rack

- ICONNMR will start executing the NMR experiments for the found samples.

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LC-MS Measurement

• Remove samples from the liquid hanlder

- Open the door of the hood.

- Remove the a rack with the SampleJet tubes.

- Close the door of the hood.

Note: If the preparation of one rack is not completely finished, the preparation will pause until the original SampleJet rack is back in the liquid handler!

• Insert the tray into the LC-MS autosampler

• Submit the ## name of the wellplate.

- The orders for this wellplates will be take from SampleTrack and will start automatically.

- It is recommended to run 1-3 blank runs before the first real sample, to equilibrate the LC-MS system.

Documents

© Bruker BioSpin 2008; File: braumann_jena_080828.ppt; Version: 2008-08-28; By: Dr. Ulrich Braumann LC-(SPE)-(Cryo)NMR(/MS) as Routine Analysis Technique: