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

From 2012.igem.org

Revision as of 13:52, 26 September 2012 by S jager (Talk | contribs)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)

1 Sample preparation
- 1.1 Purge and Trap
- 1.2 TPA short
2 Results
- 2.1 TPA Short ATS
  - 2.1.1 [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808007 Operon 2]
  - 2.1.2 [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808006http://partsregistry.org/wiki/index.php?title=Part:BBa_K808006 Operon 1]
- 2.2 Control sample

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:

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:

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

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].

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)

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

After integration of the peak ta 7 RT we obtained an area of 38700.

Control sample

The third sample is our control sample. Notably, the bacteria culture of this sample exposed an 0.85 higher OD than the samples from Operon 1 and 2 thus it will be difficult to compare.

Here we see the SCAN (first window), the SIM (second window) and the third is the mass spectra of the corresponding peak at 7 RT.