http://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/modules/methodology/sample_analysis&feed=atom&action=historyTeam:Valencia UPV/Project/modules/methodology/sample analysis - Revision history2024-03-29T04:47:15ZRevision history for this page on the wikiMediaWiki 1.16.5http://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/modules/methodology/sample_analysis&diff=378007&oldid=prevAlquiru at 00:51, 18 October 20142014-10-18T00:51:43Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>As a result a mass spectrum for each compound is obtained, i.e., the pattern of the ion fragments in which that compound breaks down, characterised by their m/z ratio and their relative abundance. This mass spectrum is characteristic for each substance and therefore a very valuable tool for compound identification.</p><br/><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>As a result a mass spectrum for each compound is obtained, i.e., the pattern of the ion fragments in which that compound breaks down, characterised by their m/z ratio and their relative abundance. This mass spectrum is characteristic for each substance and therefore a very valuable tool for compound identification.</p><br/><br/></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/9/9e/VUPV_mass.png"></img></div<del class="diffchange diffchange-inline">><br/</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/9/9e/VUPV_mass.png"></img></div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 3. </span>(Z)-11-Hexadecn-1-ol mass spectrum</p></div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 3. </span>(Z)-11-Hexadecn-1-ol mass spectrum</p></div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Source</span>. NIST Chemistry Webbook</p></div><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Source</span>. NIST Chemistry Webbook</p></div><br/></div></td></tr>
</table>Alquiruhttp://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/modules/methodology/sample_analysis&diff=377954&oldid=prevAlquiru at 00:51, 18 October 20142014-10-18T00:51:17Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/2/22/VUPVGas_chromatography_2.jpg" alt="sample_injector"></img></div><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/2/22/VUPVGas_chromatography_2.jpg" alt="sample_injector"></img></div><br/></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 1. </span>.Gas Chromatography diagram</p></div<del class="diffchange diffchange-inline">><br/</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 1. </span>.Gas Chromatography diagram</p></div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><div align="center"><p style="font-size: 0.8em; width: 70%;">K. Murray/ Wikimedia Commons / CC-BY-SA-3.0.</p></div><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><div align="center"><p style="font-size: 0.8em; width: 70%;">K. Murray/ Wikimedia Commons / CC-BY-SA-3.0.</p></div<ins class="diffchange diffchange-inline">><br/</ins>><br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Example of a chromatogram obtained by GC: each peak corresponds to a different molecule.</p><br/><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Example of a chromatogram obtained by GC: each peak corresponds to a different molecule.</p><br/><br/></div></td></tr>
</table>Alquiruhttp://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/modules/methodology/sample_analysis&diff=377876&oldid=prevAlquiru at 00:50, 18 October 20142014-10-18T00:50:43Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>Mass spectrometry is an analytical technique capable of separating charged ions according to their mass-to-charge ratio (m/z) and measuring their abundance.</p<del class="diffchange diffchange-inline">><br/</del>><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>Mass spectrometry is an analytical technique capable of separating charged ions according to their mass-to-charge ratio (m/z) and measuring their abundance.</p><br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The three basic components of a mass spectrometer are:</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The three basic components of a mass spectrometer are:</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><ul class="method"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><ul class="method"></div></td></tr>
</table>Alquiruhttp://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/modules/methodology/sample_analysis&diff=377579&oldid=prevAlquiru at 00:48, 18 October 20142014-10-18T00:48:30Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules">Modules</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology">Methodology</a> > <a>Sample Analysis GC-MS</a></h3></p><br/></br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules">Modules</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology">Methodology</a> > <a>Sample Analysis GC-MS</a></h3></p><br/></br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><div align="center"><span class="coda"><roja>S</roja>ample <roja>A</roja>nalysis<del class="diffchange diffchange-inline">. </del><roja>G</roja>C-<roja>M</roja>S</span> </div><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><div align="center"><span class="coda"><roja>S</roja>ample <roja>A</roja>nalysis<ins class="diffchange diffchange-inline">: </ins><roja>G</roja>C-<roja>M</roja>S</span> </div><br/></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p class="subpart">The Idea</p><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p class="subpart">The Idea</p><br/></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>You can find the results of the GC-MS analysis <a <del class="diffchange diffchange-inline">hrf</del>=https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis> here<del class="diffchange diffchange-inline">.</del></a<del class="diffchange diffchange-inline">></a></p><br/</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>You can find the results of the GC-MS analysis <a <ins class="diffchange diffchange-inline">href</ins>=<ins class="diffchange diffchange-inline">"</ins>https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis<ins class="diffchange diffchange-inline">" class="normal-link-page"</ins>>here</a></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>To see more details about GC-MS conditions <a class="blue-bold">see Protocol</a>.</p><br/><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>To see more details about GC-MS conditions <a class="blue-bold">see Protocol</a>.</p><br/><br/></div></td></tr>
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</table>Alquiruhttp://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/modules/methodology/sample_analysis&diff=376916&oldid=prevAlquiru at 00:43, 18 October 20142014-10-18T00:43:01Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>When it comes to analysing volatile compounds Gas Chromatography (GC) coupled to Mass spectrometry (MS) is unequivocally the first choice. The combination of <del class="diffchange diffchange-inline">both techniques allows </del>the separation and identification of each single volatile molecule present in the sample.</p><br/><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>When it comes to analysing volatile compounds Gas Chromatography (GC) coupled to Mass spectrometry (MS) is unequivocally the first choice. The combination of the separation <ins class="diffchange diffchange-inline">resolution provided by chromatography with the structural information provided by mass spectrometry allows the quantification </ins>and identification of each single volatile molecule present in the sample.</p><br/><br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"><p>First of all, the sample must be prepared and volatilized (in case the sample is not in gas state). This is not our situation, since we are analysing volatiles extracted by HS-SPME (<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/sample_preparation" class="normal-link-page">see Sample preparation - HS-SPME</a>). Once the sample is in gas state it can be introduced in the gas Chromatograph.</p><br/><br/></del></div></td><td colspan="2"> </td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p class="subpart">GAS CROMATOGRAPHY</p><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p class="subpart">GAS CROMATOGRAPHY</p><br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>As every chromatography technique, <del class="diffchange diffchange-inline">it </del>is based on <del class="diffchange diffchange-inline">the separation </del>of the components of a <del class="diffchange diffchange-inline">mixture, which is called the </del>mobile phase <del class="diffchange diffchange-inline">or eluent, when it is flowing through </del>a stationary phase.</p><br/><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>As every chromatography technique, <ins class="diffchange diffchange-inline">gas chromatography </ins>is based on <ins class="diffchange diffchange-inline">differential partitioning </ins>of the components of a <ins class="diffchange diffchange-inline">sample between a </ins>mobile phase <ins class="diffchange diffchange-inline">that acts as sample carrier (</ins>a <ins class="diffchange diffchange-inline">pure gas such as N2, He or H2 in the case of gas chromatography) and the </ins>stationary phase <ins class="diffchange diffchange-inline">coating the chromatography column</ins>.</p><br/><br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>The molecules <del class="diffchange diffchange-inline">of </del>the mixture are separated by selective retention<del class="diffchange diffchange-inline">, it is to say, if they have more </del>affinity for the mobile phase<del class="diffchange diffchange-inline">, they </del>will flow faster <del class="diffchange diffchange-inline">while if they have </del>higher affinity for the stationary phase<del class="diffchange diffchange-inline">, they </del>will <del class="diffchange diffchange-inline">be more retained by it. Therefore molecules will last different times flowing </del>through the <del class="diffchange diffchange-inline">stationary phase, it is called </del>retention time.</p><br/><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>The molecules <ins class="diffchange diffchange-inline">in </ins>the mixture are separated <ins class="diffchange diffchange-inline">as they flow through the column </ins>by selective retention<ins class="diffchange diffchange-inline">. Molecules with higher </ins>affinity for the mobile phase will flow faster <ins class="diffchange diffchange-inline">and elute the column first, whereas those with </ins>higher affinity for the stationary phase will <ins class="diffchange diffchange-inline">take longer to pass </ins>through the <ins class="diffchange diffchange-inline">system. The </ins>retention time <ins class="diffchange diffchange-inline">of a particular substance (the time it takes to pass through the column) depends on the type of column used, its length, and set temperature</ins>. </p><br/><br/></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/4/47/VUPVGas_chromatography_1.jpg" alt="analytes"></img></div><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/4/47/VUPVGas_chromatography_1.jpg" alt="analytes"></img></div><br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p><del class="diffchange diffchange-inline">In the case of Gas chromatography, the mobile phase is composed of pure gases that act as carriers, N2, He or H. The stationary phase is a capillary column with a inner hollow where the mobile phase flows through. </del>The <del class="diffchange diffchange-inline">inner surface </del>of the <del class="diffchange diffchange-inline">column </del>is <del class="diffchange diffchange-inline">coated with </del>the <del class="diffchange diffchange-inline">stationary material. The column is inside an oven </del>where <del class="diffchange diffchange-inline">the temperature </del>is <del class="diffchange diffchange-inline">raised in order to increase </del>the <del class="diffchange diffchange-inline">volatility of the analytes, decreasing the analysis time without losing resolution.</p><br/><br/></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>The <ins class="diffchange diffchange-inline">last part </ins>of the <ins class="diffchange diffchange-inline">chromatograph </ins>is the <ins class="diffchange diffchange-inline">detector </ins>where <ins class="diffchange diffchange-inline">a signal </ins>is <ins class="diffchange diffchange-inline">registered as </ins>the <ins class="diffchange diffchange-inline">compounds elute from </ins>the column. <ins class="diffchange diffchange-inline">In GC-MS </ins>the <ins class="diffchange diffchange-inline">mass spectrometer acts as </ins>detector.</p><br/><br/></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"><p>The retention characteristics of </del>the column <del class="diffchange diffchange-inline">depend on its length, material and the temperature of the oven</del>.<del class="diffchange diffchange-inline"></p><br/><br/></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"><p>As the analytes flow through the column and become separated, they arrive at </del>the detector <del class="diffchange diffchange-inline">where they will be identified</del>.</p><br/><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/2/22/VUPVGas_chromatography_2.jpg" alt="sample_injector"></img></div><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/2/22/VUPVGas_chromatography_2.jpg" alt="sample_injector"></img></div><br/></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"><div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 1. </span>.Gas Chromatography diagram</p></div><br/></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div align="center"><p style="font-size: 0.8em; width: 70%;">K. Murray/ Wikimedia Commons / CC-BY-SA-3.0.</p></div><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div align="center"><p style="font-size: 0.8em; width: 70%;">K. Murray/ Wikimedia Commons / CC-BY-SA-3.0.</p></div><br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>Example of <ins class="diffchange diffchange-inline">a chromatogram obtained </ins>by GC: <ins class="diffchange diffchange-inline">each peak corresponds to a different molecule.</ins></p><br/><br/></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>Example of <del class="diffchange diffchange-inline">different molecules separated </del>by GC:</p><br/><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/b/bf/Gas_chromatography_3_2.jpg" alt="molecules_gc"></img></div><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/b/bf/Gas_chromatography_3_2.jpg" alt="molecules_gc"></img></div><br/></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 2. </span>.Chromatogram obtained by Gas chromatography</p></div><br/><br/></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br/><p class="subpart">MASS SPECTROMETRY</p><br/><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br/><p class="subpart">MASS SPECTROMETRY</p><br/><br/></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p><del class="diffchange diffchange-inline">Once the analytes arrive at the detector, they can be identified by mass </del>spectrometry<del class="diffchange diffchange-inline">. This </del>technique <del class="diffchange diffchange-inline">works by ionizing the analytes coming from the GC </del>and measuring <del class="diffchange diffchange-inline">the </del>abundance <del class="diffchange diffchange-inline">of the formed ions</del>.</p><br/><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p><ins class="diffchange diffchange-inline">Mass </ins>spectrometry <ins class="diffchange diffchange-inline">is an analytical </ins>technique <ins class="diffchange diffchange-inline">capable of separating charged ions according to their mass-to-charge ratio (m/z) </ins>and measuring <ins class="diffchange diffchange-inline">their </ins>abundance.</p><br/><br/<ins class="diffchange diffchange-inline">></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>The three basic components of a mass spectrometer are:</p></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><ul class="method"</ins>></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><<del class="diffchange diffchange-inline">p</del>>The <del class="diffchange diffchange-inline">first step is to ionize the analytes. There are many methods but the one we used was Electron Ionization (EI). This basically works by bombing the molecules with high-energy electrons (70eV)</del>, <del class="diffchange diffchange-inline">which break the molecules into charged fragments of a range of different masses, which are characteristic for each compound</del>.</<del class="diffchange diffchange-inline">p</del>><<del class="diffchange diffchange-inline">br</del>/><<del class="diffchange diffchange-inline">br</del>/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> </ins><<ins class="diffchange diffchange-inline">li</ins>>The <ins class="diffchange diffchange-inline">ion source</ins>, <ins class="diffchange diffchange-inline">where ionization takes place</ins>. </<ins class="diffchange diffchange-inline">li</ins>></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> <li>The mass analyser.</ins></<ins class="diffchange diffchange-inline">li</ins>></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> <li>The detector. </ins></<ins class="diffchange diffchange-inline">li</ins>></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p><del class="diffchange diffchange-inline">Then, </del>the <del class="diffchange diffchange-inline">resulting fragments can be analysed according </del>to <del class="diffchange diffchange-inline">their mass/charge </del>(<del class="diffchange diffchange-inline">m/z</del>) <del class="diffchange diffchange-inline">ratio. There are many different </del>mass <del class="diffchange diffchange-inline">analysers; the quadrupole </del>analyser <del class="diffchange diffchange-inline">is the chosen one in this case. It consists of four cylindrical rods</del>, <del class="diffchange diffchange-inline">two of them having positive electric potential while the other two are negatively charged. A radio frequency voltage is applied between the rod pairs creating </del>an <del class="diffchange diffchange-inline">oscillating electric field. Only the ions with a given m/z will maintain its trajectory and cross the quadrupole to reach the </del>detector<del class="diffchange diffchange-inline">, while the rest will be deflected</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"></ul></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p><ins class="diffchange diffchange-inline">MS systems differ in </ins>the <ins class="diffchange diffchange-inline">methods used </ins>to <ins class="diffchange diffchange-inline">generate and separate the ions. Our MS system employs electron ionization </ins>(<ins class="diffchange diffchange-inline">EI</ins>)<ins class="diffchange diffchange-inline">, a quadrupole </ins>mass analyser, <ins class="diffchange diffchange-inline">and </ins>an <ins class="diffchange diffchange-inline">electron multiplier </ins>detector.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></p><br/><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></p><br/><br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p><del class="diffchange diffchange-inline">Finally</del>, <del class="diffchange diffchange-inline">a detector can define </del>the <del class="diffchange diffchange-inline">amount </del>of <del class="diffchange diffchange-inline">ions with </del>a <del class="diffchange diffchange-inline">given </del>m/z. <del class="diffchange diffchange-inline">There are also many types </del>of <del class="diffchange diffchange-inline">detectors. In this case</del>, <del class="diffchange diffchange-inline">we used </del>an <del class="diffchange diffchange-inline">electron multiplier</del>. <del class="diffchange diffchange-inline">These detectors can amplify </del>the <del class="diffchange diffchange-inline">signal of </del>a given <del class="diffchange diffchange-inline">ion into an electronic current</del>, <del class="diffchange diffchange-inline">like </del>a <del class="diffchange diffchange-inline">cascade</del>. The more quantity of ions that arrive, the greater the electron current produced. Therefore, the system is capable of quantifying the arriving ions by measuring the produced electric signal.</p><br/><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p><ins class="diffchange diffchange-inline"><b>How it works</b></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"></p><br/></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><ul class="method"></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> <li><b>Ionization:</b> </ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">The different compounds in the sample mixture enter the ionization source as they elute form the column. There</ins>, <ins class="diffchange diffchange-inline">they are bombarded with high-energy electrons (70eV), which break </ins>the <ins class="diffchange diffchange-inline">molecules into charged fragments </ins>of a <ins class="diffchange diffchange-inline">range of different masses, which are characteristic for each compound.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"></li><br/></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> <li><b>Analysis:</b> </ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">The resulting fragments are separated according to their </ins>m/z <ins class="diffchange diffchange-inline">ratio in the quadrupole analyser</ins>. <ins class="diffchange diffchange-inline">A quadrupole consists </ins>of <ins class="diffchange diffchange-inline">four cylindrical rods</ins>, <ins class="diffchange diffchange-inline">two of them having positive electric potential while the other two are negatively charged. A radio frequency voltage is applied between the rod pairs creating </ins>an <ins class="diffchange diffchange-inline">oscillating electric field</ins>. <ins class="diffchange diffchange-inline">Only </ins>the <ins class="diffchange diffchange-inline">ions with </ins>a given <ins class="diffchange diffchange-inline">m/z will maintain its trajectory and cross the quadrupole to reach the detector</ins>, <ins class="diffchange diffchange-inline">while the rest will be deflected. The voltage applied can be continuously changed (full scan) to monitor </ins>a <ins class="diffchange diffchange-inline">range of m/z values, or it can be set to monitor only specific m/z ions (single ion monitoring mode, SIM)</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"></li><br/></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"> <li><b>Detection:</b></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">The ions that cross the analyzer reached the detector which converts ions to electric currency</ins>. The more quantity of ions that arrive, the greater the electron current produced. Therefore, the system is capable of quantifying the arriving ions by measuring the produced electric signal<ins class="diffchange diffchange-inline">.</ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"></li><br/></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"></ul></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p>As a result a mass spectrum for each compound is obtained, i.e., the pattern of the ion fragments in which that compound breaks down, characterised by their m/z ratio and their relative abundance. This mass spectrum is characteristic for each substance and therefore a very valuable tool for compound identification</ins>.</p><br/><br/></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/9/9e/VUPV_mass.png"></img></div><br/></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 3. </span>(Z)-11-Hexadecn-1-ol mass spectrum</p></div></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Source</span>. NIST Chemistry Webbook</p></div><br/></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"><p>After the entire analysis process, the pheromone analysis results were obtained (<a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis" class="normal-link-page">see Results: pheromone analysis</a>)</p><br/><br/></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"><p>You can find the results of the GC-MS analysis <a hrf=https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis> here.</a></a></p><br/></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>To see more details about GC-MS conditions <a class="blue-bold">see Protocol</a>.</p><br/><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>To see more details about GC-MS conditions <a class="blue-bold">see Protocol</a>.</p><br/><br/></div></td></tr>
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</table>Alquiruhttp://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/modules/methodology/sample_analysis&diff=332077&oldid=prevIvllobel at 16:31, 17 October 20142014-10-17T16:31:21Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules">Modules</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology">Methodology</a> > <a>Sample Analysis GC-MS</a></h3></p><br/></br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules">Modules</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology">Methodology</a> > <a>Sample Analysis GC-MS</a></h3></p><br/></br></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><div align="center"><span class="coda"><roja>S</roja>ample <roja>A</roja>nalysis. <roja>G</roja>C - <roja>M</roja>S</span> </div><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><div align="center"><span class="coda"><roja>S</roja>ample <roja>A</roja>nalysis. <roja>G</roja>C-<roja>M</roja>S</span> </div><br/></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p class="subpart">The Idea</p><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p class="subpart">The Idea</p><br/></div></td></tr>
</table>Ivllobelhttp://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/modules/methodology/sample_analysis&diff=332054&oldid=prevIvllobel at 16:31, 17 October 20142014-10-17T16:31:04Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules">Modules</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology">Methodology</a> > <a>Sample Analysis <del class="diffchange diffchange-inline">CMSP</del></a></h3></p><br/></br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules">Modules</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology">Methodology</a> > <a>Sample Analysis <ins class="diffchange diffchange-inline">GC-MS</ins></a></h3></p><br/></br></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><div align="center"><span class="coda"><roja>S</roja>ample <roja>A</roja>nalysis. <roja>G</roja><del class="diffchange diffchange-inline">as</del><roja><del class="diffchange diffchange-inline">C</del></roja><del class="diffchange diffchange-inline">MSP</del></span> </div><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><div align="center"><span class="coda"><roja>S</roja>ample <roja>A</roja>nalysis. <roja>G</roja><ins class="diffchange diffchange-inline">C - </ins><roja><ins class="diffchange diffchange-inline">M</ins></roja><ins class="diffchange diffchange-inline">S</ins></span> </div><br/></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p class="subpart">The Idea</p><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p class="subpart">The Idea</p><br/></div></td></tr>
</table>Ivllobelhttp://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/modules/methodology/sample_analysis&diff=267059&oldid=prevIvllobel at 08:42, 16 October 20142014-10-16T08:42:20Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/sample_preparation"><strong>&larr; Go to Sample Preparation</strong></a></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/sample_preparation"><strong>&larr; Go to Sample Preparation</strong></a></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a class="button-content" id="goto-middle" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology"><strong>Go back to Methodology</strong></a></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a class="button-content" id="goto-middle" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology"><strong>Go back to Methodology</strong></a></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/<del class="diffchange diffchange-inline">biosafety</del>"><strong>Go to <del class="diffchange diffchange-inline">Biosafety </del>&rarr;</strong></a></div></br></br></br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/<ins class="diffchange diffchange-inline">methodology/dynamic_headspace</ins>"><strong>Go to <ins class="diffchange diffchange-inline">Sampling Technique </ins>&rarr;</strong></a></div></br></br></br></div></td></tr>
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</table>Ivllobelhttp://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/modules/methodology/sample_analysis&diff=247530&oldid=prevIvllobel at 15:12, 15 October 20142014-10-15T15:12:23Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>When it comes to analysing volatile compounds Gas Chromatography (GC) coupled to Mass spectrometry (MS) is unequivocally the first choice. The combination of both techniques allows the separation and identification of each single volatile molecule present in the sample.</p><br/><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>When it comes to analysing volatile compounds Gas Chromatography (GC) coupled to Mass spectrometry (MS) is unequivocally the first choice. The combination of both techniques allows the separation and identification of each single volatile molecule present in the sample.</p><br/><br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>First of all, the sample must be prepared and volatilized (in case the sample is not in gas state). This is not our situation, since we are analysing volatiles extracted by HS-SPME (<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/sample_preparation" class="<del class="diffchange diffchange-inline">blue</del>-<del class="diffchange diffchange-inline">bold</del>">see Sample preparation - HS-SPME</a>). Once the sample is in gas state it can be introduced in the gas Chromatograph.</p><br/><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>First of all, the sample must be prepared and volatilized (in case the sample is not in gas state). This is not our situation, since we are analysing volatiles extracted by HS-SPME (<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/sample_preparation" class="<ins class="diffchange diffchange-inline">normal</ins>-<ins class="diffchange diffchange-inline">link-page</ins>">see Sample preparation - HS-SPME</a>). Once the sample is in gas state it can be introduced in the gas Chromatograph.</p><br/><br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p class="subpart">GAS CROMATOGRAPHY</p><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p class="subpart">GAS CROMATOGRAPHY</p><br/></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Finally, a detector can define the amount of ions with a given m/z. There are also many types of detectors. In this case, we used an electron multiplier. These detectors can amplify the signal of a given ion into an electronic current, like a cascade. The more quantity of ions that arrive, the greater the electron current produced. Therefore, the system is capable of quantifying the arriving ions by measuring the produced electric signal.</p><br/><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Finally, a detector can define the amount of ions with a given m/z. There are also many types of detectors. In this case, we used an electron multiplier. These detectors can amplify the signal of a given ion into an electronic current, like a cascade. The more quantity of ions that arrive, the greater the electron current produced. Therefore, the system is capable of quantifying the arriving ions by measuring the produced electric signal.</p><br/><br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>After the entire analysis process, the pheromone analysis results were obtained (<a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis" class="<del class="diffchange diffchange-inline">blue</del>-<del class="diffchange diffchange-inline">bold</del>">see Results: pheromone analysis</a>)</p><br/><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>After the entire analysis process, the pheromone analysis results were obtained (<a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis" class="<ins class="diffchange diffchange-inline">normal</ins>-<ins class="diffchange diffchange-inline">link-page</ins>">see Results: pheromone analysis</a>)</p><br/><br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>To see more details about GC-MS conditions <a class="blue-bold">see Protocol</a>.</p><br/><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>To see more details about GC-MS conditions <a class="blue-bold">see Protocol</a>.</p><br/><br/></div></td></tr>
</table>Ivllobelhttp://2014.igem.org/wiki/index.php?title=Team:Valencia_UPV/Project/modules/methodology/sample_analysis&diff=247519&oldid=prevIvllobel at 15:11, 15 October 20142014-10-15T15:11:58Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>When it comes to analysing volatile compounds Gas Chromatography (GC) coupled to Mass spectrometry (MS) is unequivocally the first choice. The combination of both techniques allows the separation and identification of each single volatile molecule present in the sample.</p><br/><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>When it comes to analysing volatile compounds Gas Chromatography (GC) coupled to Mass spectrometry (MS) is unequivocally the first choice. The combination of both techniques allows the separation and identification of each single volatile molecule present in the sample.</p><br/><br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>First of all, the sample must be prepared and volatilized (in case the sample is not in gas state). This is not our situation, since we are analysing volatiles extracted by HS-SPME (<a class="blue-bold">see Sample preparation - HS-SPME</a>). Once the sample is in gas state it can be introduced in the gas Chromatograph.</p><br/><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>First of all, the sample must be prepared and volatilized (in case the sample is not in gas state). This is not our situation, since we are analysing volatiles extracted by HS-SPME (<a <ins class="diffchange diffchange-inline">href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/sample_preparation" </ins>class="blue-bold">see Sample preparation - HS-SPME</a>). Once the sample is in gas state it can be introduced in the gas Chromatograph.</p><br/><br/></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p class="subpart">GAS CROMATOGRAPHY</p><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p class="subpart">GAS CROMATOGRAPHY</p><br/></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>Once the <del class="diffchange diffchange-inline">gas molecules </del>arrive at the detector, they can be identified by mass spectrometry. This technique works by ionizing the <del class="diffchange diffchange-inline">gas molecules </del>coming from the GC and measuring the <del class="diffchange diffchange-inline">relative </del>abundance of the formed ions.</p><br/><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>Once the <ins class="diffchange diffchange-inline">analytes </ins>arrive at the detector, they can be identified by mass spectrometry. This technique works by ionizing the <ins class="diffchange diffchange-inline">analytes </ins>coming from the GC and measuring the abundance of the formed ions.</p><br/><br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>The first step is to ionize the <del class="diffchange diffchange-inline">gas molecules</del>. There are many methods but the one used was Electron Ionization (EI). This basically works by bombing the molecules with high-energy electrons <del class="diffchange diffchange-inline">until they </del>break into fragments of different <del class="diffchange diffchange-inline">charge and mass</del>.</p><br/><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>The first step is to ionize the <ins class="diffchange diffchange-inline">analytes</ins>. There are many methods but the one <ins class="diffchange diffchange-inline">we </ins>used was Electron Ionization (EI). This basically works by bombing the molecules with high-energy electrons <ins class="diffchange diffchange-inline">(70eV), which </ins>break <ins class="diffchange diffchange-inline">the molecules </ins>into <ins class="diffchange diffchange-inline">charged </ins>fragments <ins class="diffchange diffchange-inline">of a range </ins>of different <ins class="diffchange diffchange-inline">masses, which are characteristic for each compound</ins>.</p><br/><br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>Then, the resulting fragments <del class="diffchange diffchange-inline">must </del>be analysed according to their mass/charge (m/z) ratio. There are many different mass analysers; the quadrupole analyser is the chosen one in this case. It consists of four cylindrical rods, two of them having positive electric potential while the other two are negatively charged. A radio frequency voltage is applied between the rod pairs creating an oscillating electric field. Only the ions with a given m/z will maintain its trajectory and cross the quadrupole to <del class="diffchange diffchange-inline">be detected</del>, the rest will be deflected.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>Then, the resulting fragments <ins class="diffchange diffchange-inline">can </ins>be analysed according to their mass/charge (m/z) ratio. There are many different mass analysers; the quadrupole analyser is the chosen one in this case. It consists of four cylindrical rods, two of them having positive electric potential while the other two are negatively charged. A radio frequency voltage is applied between the rod pairs creating an oscillating electric field. Only the ions with a given m/z will maintain its trajectory and cross the quadrupole to <ins class="diffchange diffchange-inline">reach the detector</ins>, <ins class="diffchange diffchange-inline">while </ins>the rest will be deflected.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></p><br/><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></p><br/><br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>Finally, a detector can define the amount of ions with a given m/<del class="diffchange diffchange-inline">r</del>. There are also many types of detectors. In this case, an electron multiplier <del class="diffchange diffchange-inline">was used</del>. These detectors can amplify the signal of a given ion into an electronic current, like a cascade. The more quantity of ions that arrive, the greater the electron current. Therefore, the system is capable of quantifying the arriving ions by measuring the produced electric signal.</p><br/><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>Finally, a detector can define the amount of ions with a given m/<ins class="diffchange diffchange-inline">z</ins>. There are also many types of detectors. In this case, <ins class="diffchange diffchange-inline">we used </ins>an electron multiplier. These detectors can amplify the signal of a given ion into an electronic current, like a cascade. The more quantity of ions that arrive, the greater the electron current <ins class="diffchange diffchange-inline">produced</ins>. Therefore, the system is capable of quantifying the arriving ions by measuring the produced electric signal.</p><br/><br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>After <del class="diffchange diffchange-inline">all </del>the analysis <del class="diffchange diffchange-inline">project</del>, the pheromone analysis results were obtained (<a class="blue-bold">see Results: pheromone analysis</a>)</p><br/><br/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>After the <ins class="diffchange diffchange-inline">entire </ins>analysis <ins class="diffchange diffchange-inline">process</ins>, the pheromone analysis results were obtained (<a <ins class="diffchange diffchange-inline">href="https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis" </ins>class="blue-bold">see Results: pheromone analysis</a>)</p><br/><br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>To see more details about GC-MS conditions <a class="blue-bold">see Protocol</a>.</p><br/><br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>To see more details about GC-MS conditions <a class="blue-bold">see Protocol</a>.</p><br/><br/></div></td></tr>
</table>Ivllobel