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class="content"><article class="post-15505 post type-post status-publish format-standard has-post-thumbnail category-gc-ms tag-food-contaminants tag-mosh-moah entry" aria-label="Headspace GC-MS Analysis: Ideal for Rancidity in Nuts"><header class="entry-header"><h1 class="entry-title">Headspace GC-MS Analysis: Ideal for Rancidity in Nuts</h1> <span class="post-info-author"> By <a href="https://www.thermofisher.com/blog/analyteguru/tf-author/petra-gerhards/" title="Posts by Petra Gerhards" rel="author">Petra Gerhards, </a> </span> <span class="post-info-date"> 04.24.2023 </span> </header><div class="entry-content"> <p><em>Editor’s note: Contributing to this blog post were </em><em>Timm Sperling, Technical Sales Specialist, </em><em>Analytical Instruments Group, Dreieich Germany; a</em><em>nd Dr. Thomas Stegemann,<strong> </strong>Sr. Field Application Scientist GC, GC-MS, Dreieich Germany.</em></p> <figure class="wp-block-gallery alignright has-nested-images columns-default is-cropped wp-block-gallery-1 is-layout-flex wp-block-gallery-is-layout-flex"> <figure class="wp-block-image size-large"><img decoding="async" width="1000" height="588" data-id="10496" src="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2023/06/Detecting-rancid-off-flavors-in-nuts.jpg" alt="Walnuts" class="wp-image-10496" srcset="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2023/06/Detecting-rancid-off-flavors-in-nuts.jpg 1000w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2023/06/Detecting-rancid-off-flavors-in-nuts-300x176.jpg 300w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2023/06/Detecting-rancid-off-flavors-in-nuts-768x452.jpg 768w" sizes="(max-width: 1000px) 100vw, 1000px" /></figure> </figure> <p class="has-text-align-left">Nuts are some of the most prized foods from among nature’s bounty. Nuts are endowed with healthy, polyunsaturated and monounsaturated fats. But what makes nuts so healthy is also what makes them perishable — the unsaturated fats. Nuts have a longer shelf life than our fresh fruits and vegetables, but they are susceptible to rancidity over a period of time. Rancidity takes place when the fat inside nuts <a href="https://food.ndtv.com/food-drinks/rancidity-in-nuts-how-to-tell-when-nuts-are-stale-or-rancid-2023870" target="_blank" rel="noreferrer noopener">becomes oxidized</a>.</p> <p>Rancidification is the process of complete or incomplete <a href="https://en.wikipedia.org/wiki/Autoxidation" target="_blank" rel="noreferrer noopener">autoxidation</a> or <a href="https://en.wikipedia.org/wiki/Hydrolysis" target="_blank" rel="noreferrer noopener">hydrolysis</a> of <a href="https://en.wikipedia.org/wiki/Fat" target="_blank" rel="noreferrer noopener">fats</a> and oils when exposed to air, light, moisture, or bacterial action, producing short-chain <a href="https://en.wikipedia.org/wiki/Aldehydes" target="_blank" rel="noreferrer noopener">aldehydes</a>, <a href="https://en.wikipedia.org/wiki/Ketones" target="_blank" rel="noreferrer noopener">ketones</a> and free fatty acids.</p> <p>For this blog post, fresh walnuts were measured and compared to rancid walnuts that were past their expiration date. The analysis was performed comparing a <a href="https://www.thermofisher.com/de/en/home/industrial/mass-spectrometry/gas-chromatography-mass-spectrometry-gc-ms/gc-ms-systems/single-quadrupole-gc-ms.html?gclid=EAIaIQobChMIkri_veTx_QIVP4xoCR1bDAHmEAAYASAAEgJ6W_D_BwE&cid=E.23CMD.DL103.12921.01&ef_id=EAIaIQobChMIkri_veTx_QIVP4xoCR1bDAHmEAAYASAAEgJ6W_D_BwE:G:s&s_kwcid=AL!3652!3!634270944213!p!!g!!isq%207610" target="_blank" rel="noreferrer noopener">Thermo Scientific <sup>TM</sup> ISQ 7610</a> and a <a href="https://www.thermofisher.com/de/en/home/industrial/mass-spectrometry/gas-chromatography-mass-spectrometry-gc-ms/gc-ms-systems/high-resolution-accurate-mass-gc-ms.html?gclid=EAIaIQobChMIlIeS7eTx_QIVB5vVCh0JOgUaEAAYASAAEgLvLvD_BwE&cid=E.23CMD.DL103.12921.01&ef_id=EAIaIQobChMIlIeS7eTx_QIVB5vVCh0JOgUaEAAYASAAEgLvLvD_BwE:G:s&s_kwcid=AL!3652!3!634270944228!p!!g!!exploris%20gc" target="_blank" rel="noreferrer noopener">Thermo Scientific Orbitrap™ Exploris</a>.</p> <h3 class="wp-block-heading" id="toc-hId-1155121396"><strong>Which technology can be used to detect rancid off-flavors?</strong></h3> <p>The sample introduction is done using static headspace technology with a Thermo Scientific <a href="https://www.thermofisher.com/order/catalog/product/25118200" target="_blank" rel="noreferrer noopener">TriPlus™ 500  Headspace Autosampler</a> on the ISQ 7610 and a Thermo Scientific <a href="https://www.thermofisher.com/order/catalog/product/1R77010-2001?gclid=EAIaIQobChMIwKfdh-Xx_QIVGMrVCh3Ehg4_EAAYASAAEgLDdvD_BwE&cid=E.23CMD.DL103.12921.01&ef_id=EAIaIQobChMIwKfdh-Xx_QIVGMrVCh3Ehg4_EAAYASAAEgLDdvD_BwE:G:s&s_kwcid=AL!3652!3!612652239553!p!!g!!triplus%20rsh" target="_blank" rel="noreferrer noopener">TriPlus RSH</a> autosampler on the Orbitrap Exploris. Figure 1 shows the different walnut samples in headspace vials.</p> <figure class="wp-block-image size-large"><img decoding="async" width="1024" height="417" src="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Walnut-samples-in-headspace-vials-2-1024x417.jpg" alt="" class="wp-image-16430" srcset="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Walnut-samples-in-headspace-vials-2-1024x417.jpg 1024w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Walnut-samples-in-headspace-vials-2-300x122.jpg 300w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Walnut-samples-in-headspace-vials-2-768x313.jpg 768w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Walnut-samples-in-headspace-vials-2.jpg 1153w" sizes="(max-width: 1024px) 100vw, 1024px" /></figure> <p><em>Figure 1: Walnut samples in headspace vials.</em></p> <p>The separation was performed on a <a href="https://www.thermofisher.com/order/catalog/product/MI-148000-0001?gclid=EAIaIQobChMIiv7TouXx_QIVRafVCh3MsQzZEAAYASAAEgI5T_D_BwE&cid=E.23CMD.DL103.12921.01&ef_id=EAIaIQobChMIiv7TouXx_QIVRafVCh3MsQzZEAAYASAAEgI5T_D_BwE:G:s&s_kwcid=AL!3652!3!443892198157!p!!g!!trace%201600" target="_blank" rel="noreferrer noopener">Thermo Scientific TRACE™ 1610</a> gas chromatograph. Detection depends on the amount of sensitivity you would like. For this blog post, an ISQ 7610 was used and an Orbitrap Exploris.</p> <figure class="wp-block-image size-full is-resized"><img loading="lazy" decoding="async" width="935" height="341" src="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/042323-Table.jpg" alt="" class="wp-image-16458" style="width:849px;height:auto" srcset="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/042323-Table.jpg 935w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/042323-Table-300x109.jpg 300w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/042323-Table-768x280.jpg 768w" sizes="(max-width: 935px) 100vw, 935px" /></figure> <h3 class="wp-block-heading" id="toc-hId--1364838538"><strong>Sample measurements</strong></h3> <p>The goal here was to find relevant substances for fresh and rancid nuts. In addition, a comparison to the Orbitrap Exploris with a 60k resolution was performed. Before every series on both instruments a blank was run. Figure 2 shows the measured samples and the order, there were measured in on the ISQ 7610.</p> <figure class="wp-block-gallery has-nested-images columns-default is-cropped wp-block-gallery-2 is-layout-flex wp-block-gallery-is-layout-flex"> <figure class="wp-block-image size-full"><img loading="lazy" decoding="async" width="678" height="373" data-id="16447" src="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Nut-samples.png" alt="" class="wp-image-16447" srcset="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Nut-samples.png 678w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Nut-samples-300x165.png 300w" sizes="(max-width: 678px) 100vw, 678px" /></figure> </figure> <p><em>Figure 2: Nut samples.</em></p> <p>Figure 3 shows a comparison of all four nut samples in total-ion-chromatogram (TIC) and on the right side a comparison of a fresh sample and an expired sample.</p> <figure class="wp-block-image size-full is-resized"><img loading="lazy" decoding="async" width="756" height="425" src="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Figure-3-TIC-of-walnut-samples.png" alt="" class="wp-image-16433" style="width:858px;height:auto" srcset="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Figure-3-TIC-of-walnut-samples.png 756w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Figure-3-TIC-of-walnut-samples-300x169.png 300w" sizes="(max-width: 756px) 100vw, 756px" /></figure> <p><em>Figure 3: TIC of all samples, blank and fresh/rancid sample.</em></p> <p>Already the first glance of the TIC shows the differences of the samples. There are characteristic peaks for fresh, as well as rancid sample. The TIC on the right also visualizes the differences. In the rancid sample, <a href="https://www.thermofisher.com/order/catalog/product/de/de/CHROMELEON7" target="_blank" rel="noreferrer noopener">Thermo Scientific  Chromeleon</a>™ CDS identified 53 peaks in total via NIST library search. Some examples of off-flavors for rancid are shown in Figure 4. As expected, those were ketones and ketals.</p> <figure class="wp-block-image size-full is-resized"><img loading="lazy" decoding="async" width="757" height="403" src="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Pantonol-Pentanal-Hexanal-and-Octenal.png" alt="" class="wp-image-16437" style="width:842px;height:auto" srcset="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Pantonol-Pentanal-Hexanal-and-Octenal.png 757w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Pantonol-Pentanal-Hexanal-and-Octenal-300x160.png 300w" sizes="(max-width: 757px) 100vw, 757px" /></figure> <p><em>Figure 4: 1=1-Pantanol; 2=Pentanal; 3=Hexanal; 4=2-Octenal.</em></p> <p> Figure 5 shows all 53 identified compounds on the ISQ.</p> <figure class="wp-block-image size-full is-resized"><img loading="lazy" decoding="async" width="844" height="475" src="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/NIST-Library-Seaerch-Results.png" alt="" class="wp-image-16438" style="width:857px;height:auto" srcset="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/NIST-Library-Seaerch-Results.png 844w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/NIST-Library-Seaerch-Results-300x169.png 300w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/NIST-Library-Seaerch-Results-768x432.png 768w" sizes="(max-width: 844px) 100vw, 844px" /></figure> <p><em>Figure 5: NIST library search results.</em></p> <p>The same samples were now measured on an Orbitrap Exploris in triplicate. Figure 6 shows the TIC on the Orbitrap Exploris. Using Chromeleon CDS for interpretation of the results, only 19 compounds could be identified.</p> <figure class="wp-block-image size-full is-resized"><img loading="lazy" decoding="async" width="605" height="504" src="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/TIC-on-the-Orbitrap-Exploris.png" alt="" class="wp-image-16439" style="width:840px;height:auto" srcset="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/TIC-on-the-Orbitrap-Exploris.png 605w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/TIC-on-the-Orbitrap-Exploris-300x250.png 300w" sizes="(max-width: 605px) 100vw, 605px" /></figure> <p><em>Figure 6: TIC on the Orbitrap Exploris.</em></p> <p>Now with Orbitrap HRMS data, it is possible to use a special software for Deconvolution call <a href="https://www.thermofisher.com/de/en/home/industrial/mass-spectrometry/liquid-chromatography-mass-spectrometry-lc-ms/lc-ms-software/multi-omics-data-analysis/compound-discoverer-software.html?gclid=EAIaIQobChMIkuGZ-uXx_QIV-5BoCR1IlwPpEAAYASAAEgKiU_D_BwE&cid=E.23CMD.DL103.12911.01&ef_id=EAIaIQobChMIkuGZ-uXx_QIV-5BoCR1IlwPpEAAYASAAEgKiU_D_BwE:G:s&s_kwcid=AL!3652!3!569181149181!p!!g!!compound%20discoverer" target="_blank" rel="noreferrer noopener">Thermo Scientific <sup>TM</sup> Compound Discoverer</a> (CD). Deconvolution is a key to success to really take a deeper dive into data. In addition, CD allows a statistical evaluation of the data.</p> <p>Figure 7 shows the CD workflow for the interpretation of off-flavors in walnuts.</p> <figure class="wp-block-image size-full is-resized"><img loading="lazy" decoding="async" width="390" height="434" src="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/CD-Workflow.png" alt="" class="wp-image-16440" style="width:487px;height:auto" srcset="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/CD-Workflow.png 390w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/CD-Workflow-270x300.png 270w" sizes="(max-width: 390px) 100vw, 390px" /></figure> <p><em>Figure 7: CD workflow.</em></p> <p>Figure 8 shows the identification of isopropyl alcohol after deconvolution.</p> <figure class="wp-block-image size-full is-resized"><img loading="lazy" decoding="async" width="683" height="337" src="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Overlay-and-ID-of-Isopropyl-Alcohol.png" alt="" class="wp-image-16443" style="width:841px;height:auto" srcset="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Overlay-and-ID-of-Isopropyl-Alcohol.png 683w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Overlay-and-ID-of-Isopropyl-Alcohol-300x148.png 300w" sizes="(max-width: 683px) 100vw, 683px" /></figure> <p><em>Figure 8: Overlay and identification of isopropyl alcohol.</em></p> <p>Since CD allows a statistical comparison of data, the upper left part shows an overlay of all samples. This shows clearly the isoproyl alcohol is only contained in rancid walnuts (orange chromatograms). It was identified with a total score of 96.3 percent.  The Orbitrap Exploris shows in general very low delta ppm`s which leads into a high accuracy for the identified compound.</p> <p>In addition to markers for rancid nuts, there are also markers for fresh nuts. In Figure 9 is an example shown of an overlay, where you can see a marker, that only shows up in fresh nuts.</p> <figure class="wp-block-image size-full is-resized"><img loading="lazy" decoding="async" width="605" height="253" src="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Marker-For-Fresh-Nuts-1-Heptene-and-5-methyl.png" alt="" class="wp-image-16444" style="width:879px;height:auto" srcset="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Marker-For-Fresh-Nuts-1-Heptene-and-5-methyl.png 605w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Marker-For-Fresh-Nuts-1-Heptene-and-5-methyl-300x125.png 300w" sizes="(max-width: 605px) 100vw, 605px" /></figure> <p><em>Figure 9: Marker for fresh nuts 1-Heptene, 5-methyl.</em></p> <p>In Figure 10, a principal component analysis (PCA) is shown. Here the total amount of compounds, without background, are shown. This immediately shows the differences for fresh and rancid walnuts.</p> <figure class="wp-block-image size-full is-resized"><img loading="lazy" decoding="async" width="440" height="348" src="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/PCA.png" alt="" class="wp-image-16445" style="width:809px;height:auto" srcset="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/PCA.png 440w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/PCA-300x237.png 300w" sizes="(max-width: 440px) 100vw, 440px" /></figure> <p><em>Figure 10: PCA.</em> </p> <p>A differential analysis is another way to look at the data. Figure 11 shows such a comparison in a volcano plot. </p> <p>What does a volcano plot show?</p> <ul class="wp-block-list"> <li>What is inside the squares is relevant</li> <li>In the pink field you see the relevant compound for fresh walnuts</li> <li>In the green field you see the relevant compounds for rancid walnuts</li> <li>The P value describes the statistical relevance</li> <li>The Log Fold shows the amount, concentration</li> <li>Below the squares, the background is shown</li> </ul> <figure class="wp-block-image size-full is-resized"><img loading="lazy" decoding="async" width="605" height="470" src="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Volcano-Plot.png" alt="" class="wp-image-16446" style="width:834px;height:auto" srcset="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Volcano-Plot.png 605w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2024/01/Volcano-Plot-300x233.png 300w" sizes="(max-width: 605px) 100vw, 605px" /></figure> <p><em>Figure 11: Volcano plot.</em></p> <p>Using the Orbitrap Exploris 265 compounds could be identified after deconvolution. This shows the power of HRMS in combination with CD.</p> <h3 class="wp-block-heading" id="toc-hId-410168824"><strong>Conclusions</strong></h3> <p>As this example shows, GC-MS, in combination with headspace, is the ideal combination to determine rancid off-flavors in walnuts. Also, it is possible to detect markers for fresh walnuts.</p> <p>Both technologies, single quadruple and HRMS, showed differences in the sample. Depending on the required information and the budget, it can be a good decision to use Orbitrap technology. As the example has shown, in combination with deconvolution, it is possible to generate much more information on an Orbitrap compared to a single quadrupole. </p> <p>Using the full power of CD, a statistical comparison allows an easy overview of compounds that are considered as rancid off-flavors. This is a fully automated integrated workflow. Hundreds of samples can be compared at the same time. Depending on the depth of statistic that is required, a lot more information can be pulled out of CD. An example with more insights to CD is linked below.</p> <p>In general, GC-MS delivers proper results to determine off-flavors in all kinds of different matrices.</p> <h3 class="wp-block-heading" id="toc-hId-1297672505"><strong>Related information</strong></h3> <p>Analyte Guru blog: <a href="https://www.thermofisher.com/blog/analyteguru/do-all-your-yogurts-taste-the-same-if-not-you-need-a-gc-orbitrap/" target="_blank" rel="noreferrer noopener">Do all Your Yogurts Taste the Same?</a></p> <p>To learn more, attend the on-demand webinar: <a href="https://event.on24.com/wcc/r/4007980/0A915BC3194513861235E0507C0C2B8D?partnerref=AnalyteGuru" target="_blank" rel="noreferrer noopener">Off-flavor analysis in food</a></p> </div></article><section class="author-box"><h4 class="author-box-title">Petra Gerhards</h4><div class="author-box-content" itemprop="description">Petra Gerhards, Dipl-Ing, is Regional Marketing Manager of GC and GC-MS for EMEA at Thermo Fisher Scientific. She has more than 29 years of experience in the fields of GC-MS, SPE and LC-MS. Since joining the regional team she has contributed to workflow solutions combining vials and closures with SPE solutions, GC-MS and LC-MS. She works with KOL's on data for regional specific marketing campaigns, organizes in-house seminars and works on customer specific solutions. Her main expertise is in the field of doping and drugs-of-abuse analysis.</div></section><div class="adjacent-entry-pagination pagination"><div class="pagination-previous alignleft"><i class="fa fa-icon fa-chevron-left"></i> <a href="https://www.thermofisher.com/blog/analyteguru/what-does-ai-really-know-about-hplc-technology/" rel="prev">What Does AI Really Know About HPLC Technology?</a></div><div class="pagination-next alignright"><a href="https://www.thermofisher.com/blog/analyteguru/aav-vectors-for-gene-therapy-top-questions-answered/" rel="next">AAV Vectors for Gene Therapy: Top Questions Answered</a> <i class="fa fa-icon fa-chevron-right"></i></div></div></main></div></div><footer class="site-footer"><div class="wrap"><div class="creds"><p class="footer-links"><a href="https://corporate.thermofisher.com/en/footer/privacy-statement.html" target="_blank" rel="noopener noreferrer">Privacy Statement</a><a href="https://corporate.thermofisher.com/en/footer/terms-and-conditions.html" target="_blank" rel="noopener noreferrer">Terms & Conditions</a><a href="/blog/locations/">Locations</a><a href="/blog/sitemap/">Sitemap</a></p><p>© 2024 <a href="https://www.thermofisher.com">Thermo Fisher Scientific</a>. 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