Team:TU Darmstadt/Protocols/GC-MS/Results

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Revision as of 12:43, 26 September 2012 by A.Schmidt (Talk | contribs)

Contents

Sample preparation

We used two methods for our TPA analytics. Therefore we used standard sample preparation protocol:

Purge and Trap

  • OP-1 add. 20 µl in 10 ml Water
  • OP-2 add. 20 µl in 10 ml Water
  • OP-K add. 20 µl in 10 ml Water

TPA short

  • OP-1 add. 1ml in vial
  • OP-2 add. 1ml in vial
  • OP-K add. 1ml in vial

Results

TPA Short ATS

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K808007 Operon 2]

The first sample we measured was [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808007 Operon 2]. Simultaneously, it was the proof of principal that our method worked. Here we present the corresponding SCAN, SIM of our sample:

OP2 full spectrum.jpg

Here we measured a biological sample hence we got a lot of peaks in the SCAN (first window). Surprisingly, in the SIM we got a huge peak at 7 minutes RT. We checked the mass spectrum out of the SCAN (third window) and obtained nearly the same fragmentation pattern we measured in the reference. Due to this, we compared the quality of our spectrum:

Op2 database structure.jpg

Here we can see that our spectrum is nearly identical to the reference spectrum of the TPA-methylester.

Op2.jpg

This screen-shot shows our SCAN and SIM (zoomed in). Due to the quantification, we need a peak with an sophisticated shape. Fortunately, the peak is perfect for the integration. After the integration we obtained a peak area of 43113.

Notably,  we are now able to quantify and qualify TPA out of biological sample via GC-MS!

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K808006http://partsregistry.org/wiki/index.php?title=Part:BBa_K808006 Operon 1]

The second sample is [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808006http://partsregistry.org/wiki/index.php?title=Part:BBa_K808006 Operon 1].

OP1 full.jpg

Here you can see the mass spectrum of our molecule as well as the SIM and the SCAN. Hence we are able to reproduce the results from our method (look at the peak at 7 RT)

OP1 mass reference.jpg

Here we illustrate the mass spectra of TPA with the corresponding reference spectrum.

OP1 mass spectrum.jpg

KO

KO mass spectrum.jpg