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class="breadcrumb-current-title">New Dimensions in Isotope Ratio Analysis: Multiply Substituted Isotopologues</span></div></div></header><div class="site-inner"><div class="content-sidebar-wrap"><main class="content"><article class="post-15901 post type-post status-publish format-standard has-post-thumbnail category-isotope-ratio-ms category-metabolomics tag-isotope-ratio-ms tag-orbitrap entry" aria-label="New Dimensions in Isotope Ratio Analysis: Multiply Substituted Isotopologues"><header class="entry-header"><h1 class="entry-title">New Dimensions in Isotope Ratio Analysis: Multiply Substituted Isotopologues</h1> <span class="post-info-author"> By <a href="https://www.thermofisher.com/blog/analyteguru/tf-author/issaku-kohl/" title="Posts by Issaku Kohl" rel="author">Issaku Kohl, </a> </span> <span class="post-info-date"> 10.13.2023 </span> </header><div class="entry-content"> <p><em>This is the second in a series on Isotope analysis using Orbitrap technology.</em></p> <p>Multiply substituted isotopologues? It may sound complicated, but it’s remarkably simple and extremely useful. And even better, it’s easy to measure with the Thermo Scientific Orbitrap Exploris Isotope Solutions.</p> <h3 class="wp-block-heading" id="h-what-is-a-multiply-substituted-isotopologue"><strong>What is a multiply substituted isotopologue?</strong></h3> <p>Take a molecule, e.g., CO<sub>2</sub>. The most common variety of CO<sub>2</sub> is <sup>12</sup>C<sup>16</sup>O<sup>16</sup>O. But in rare cases, the <sup>12</sup>C could be replaced by a <sup>13</sup>C and the <sup>16</sup>O could be replaced by <sup>17</sup>O or <sup>18</sup>O. Where two or more of these isotopes have been swapped, this is called a multiply substituted isotopologue.</p> <h3 class="wp-block-heading" id="h-why-do-we-care-about-multiply-substituted-isotopologues"><strong>Why do we care about</strong> <strong>multiply substituted isotopologues?</strong></h3> <p>Multiply substituted isotopologues give us important insights into the conditions and processes forming that molecule. For example, the carbonate multiple substituted isotopologue (<sup>13</sup>C<sup>18</sup>O<sup>16</sup>O, aka Δ47) has been widely used as a paleo-thermometer that is independent of variations in the oxygen isotopic composition of the source water. Similarly, multiply substituted isotopologues of methane have been used to identify the process of methane production in carbon capture and storage sites (e.g., <a href="https://www.nature.com/articles/s41586-021-04153-3" target="_blank" rel="noreferrer noopener">Tyne et al. 2021</a>).  Multiply substituted isotopologues can provide important insights in biochemical applications, using isotope labeling to decipher metabolic pathways.</p> <figure class="wp-block-image size-large"><img decoding="async" width="1024" height="614" src="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2023/10/HubbleBrochureCover-TR-orby-swirl-blog-2-Final-1024x614.jpg" alt="" class="wp-image-15904" srcset="https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2023/10/HubbleBrochureCover-TR-orby-swirl-blog-2-Final-1024x614.jpg 1024w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2023/10/HubbleBrochureCover-TR-orby-swirl-blog-2-Final-300x180.jpg 300w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2023/10/HubbleBrochureCover-TR-orby-swirl-blog-2-Final-768x461.jpg 768w, https://www.thermofisher.com/blog/analyteguru/wp-content/uploads/sites/25/2023/10/HubbleBrochureCover-TR-orby-swirl-blog-2-Final.jpg 1200w" sizes="(max-width: 1024px) 100vw, 1024px" /></figure> <h3 class="wp-block-heading" id="h-why-are-multiply-substituted-isotopologues-traditionally-tricky-to-analyze"><strong>Why are multiply substituted isotopologues traditionally tricky to analyze?</strong></h3> <p>Because of the difference in abundance between multiply substituted isotopologues and the major species, multiply substituted isotopologues have traditionally been tricky to measure. They require instrumentation that is sensitive enough to detect the minor isotope beams and also has the resolution to detect isobaric species, e.g., <sup>13</sup>C<sup>18</sup>O<sup>16</sup>O from <sup>12</sup>C<sup>18</sup>O<sup>17</sup>O.</p> <h3 class="wp-block-heading" id="h-what-is-game-changing-about-the-orbitrap-exploris-isotope-solutions"><strong>What is game changing about the Orbitrap Exploris Isotope Solutions?</strong></h3> <p>The Orbitrap Exploris Isotope Solutions has a mass resolution of up to 480,000 (at 200 <em>m</em>/<em>z</em>), allowing separation of isotopologues that are far beyond the reach of even the highest resolution magnetic sector mass spectrometer.  What’s more, because it is possible to limit the mass range entering the analyzer, the major species can be excluded (so-called “noM0” methodology), allowing a greater number of counts on the multiply substituted isotopologues. This means that the sensitivity of Orbitrap Exploris Isotope Solutions is adaptable for your application and the multiply substituted isotopologues of interest.</p> <p>For more information, visit: <a href="https://www.thermofisher.com/order/catalog/product/IQLAAMGAATFARBMBNP" target="_blank" rel="noreferrer noopener">thermofisher.com/orbitrap-for-isotopes</a>.</p> <p>Bulk isotopic composition of multiply substituted isotopologues is just the tip of the iceberg in terms of the advantage of the Orbitrap Exploris Isotope Solutions. Read Part 3 of this series to find out about how this technology can be used to uncover site-specific isotope information.</p> <p><strong>Related blog posts in this three-part series</strong></p> <p>Part 1: <a href="https://www.thermofisher.com/blog/analyteguru/boost-your-productivity-with-the-isotopic-analysis-of-intact-molecules/" target="_blank" rel="noreferrer noopener">Boost Your Productivity with the Isotopic Analysis of Intact Molecules</a></p> <p>Part 3: <a href="https://www.thermofisher.com/blog/analyteguru/new-dimensions-in-isotope-ratio-analysis-site-specificity/" target="_blank" rel="noreferrer noopener">New Dimensions in Isotope Ratio Analysis: Site Specificity</a></p> </div></article><section class="author-box"><h4 class="author-box-title">Issaku Kohl</h4><div class="author-box-content" itemprop="description"></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/biopharma-boost-how-2d-lc-went-from-expert-only-to-routine/" rel="prev">Biopharma Boost: How 2D-LC Went From ‘Expert Only’ to Routine</a></div><div class="pagination-next alignright"><a href="https://www.thermofisher.com/blog/analyteguru/upgrading-your-liquid-chromatography-systems/" rel="next">Upgrading Your Lab’s LC Systems? 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