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xmlns:xlink="http://www.w3.org/1999/xlink"><use xlink:href="#icon-services-invitrometabolism"></use></svg> <h1>In vitro Metabolism</h1> </div> </div> </div> </div> <main class="wp-block-group main main--page is-layout-flow wp-block-group-is-layout-flow"> <div class="wp-block-group main__content is-layout-constrained wp-block-group-is-layout-constrained"> <div id="ez-toc-container" class="ez-toc-v2_0_69_1 counter-flat ez-toc-counter ez-toc-grey ez-toc-container-direction"> <div class="ez-toc-title-container"> <p class="ez-toc-title" style="cursor:inherit">Table of Contents</p> <span class="ez-toc-title-toggle"></span></div> <nav><ul class='ez-toc-list ez-toc-list-level-1 ' ><li class='ez-toc-page-1'><a class="ez-toc-link ez-toc-heading-1" href="#Our_services_for_in_vitro_Metabolism_research" title="Our services for in vitro Metabolism research">Our services for in vitro Metabolism research</a></li><li class='ez-toc-page-1'><a class="ez-toc-link ez-toc-heading-2" href="#Metabolic_Stability" title="Metabolic Stability">Metabolic Stability</a></li><li class='ez-toc-page-1'><a class="ez-toc-link ez-toc-heading-3" href="#Metabolite_Identification_and_Profiling" title="Metabolite Identification and Profiling">Metabolite Identification and Profiling</a></li><li class='ez-toc-page-1'><a class="ez-toc-link ez-toc-heading-4" href="#Phenotyping_of_Metabolizing_Enzymes" title="Phenotyping of Metabolizing Enzymes">Phenotyping of Metabolizing Enzymes</a></li><li class='ez-toc-page-1'><a class="ez-toc-link ez-toc-heading-5" href="#Non-CYP_Mediated_Metabolism" title="Non-CYP Mediated Metabolism">Non-CYP Mediated Metabolism</a></li><li class='ez-toc-page-1'><a class="ez-toc-link ez-toc-heading-6" href="#Extrahepatic_Metabolism" title="Extrahepatic Metabolism">Extrahepatic Metabolism</a></li><li class='ez-toc-page-1'><a class="ez-toc-link ez-toc-heading-7" href="#Metabolite_Production" title="Metabolite Production">Metabolite Production</a></li><li class='ez-toc-page-1'><a class="ez-toc-link ez-toc-heading-8" href="#Reactive_Metabolites" title="Reactive Metabolites">Reactive Metabolites</a></li><li class='ez-toc-page-1'><a class="ez-toc-link ez-toc-heading-9" href="#Acyl_Glucuronide_Reactivity" title="Acyl Glucuronide Reactivity">Acyl Glucuronide Reactivity</a></li></ul></nav></div> <div class="entry-content wp-block-post-content is-layout-flow wp-block-post-content-is-layout-flow"> <h2 class="wp-block-heading"><span class="ez-toc-section" id="Our_services_for_in_vitro_Metabolism_research"></span>Our services for <em>in vitro</em> Metabolism research<span class="ez-toc-section-end"></span></h2> <p class="has-medium-font-size">Our core expertise and the origin of our services lie in drug metabolism studies, which are available for all drug modalities, e.g., small molecules, peptides, oligonucleotides and ADCs.</p> <p class="has-medium-font-size">We offer high quality <em>in vitro</em> metabolism services to evaluate the metabolic fate of your compound in the liver and in extrahepatic tissues, such as intestine, kidney, lung or skin. Assays are optimized for metabolite identification and profiling or measuring <em>in vitro</em> clearance.</p> <p class="has-medium-font-size">In vitro metabolism assays are performed using various available enzyme source, from multi-species hepatocytes to subcellular fractions and recombinant enzymes, with high quality analytical UPLC/HR-MS-instrumentation. Studies for screening reactive drug metabolites and evaluation of acyl-glucuronide conjugate reactivity are included, as well as elucidation of the enzymes involved in the metabolism of your compound (CYP/UGT/SULT/NAT/FMO/MAO/CE/ALDH/AOX/XO/AKR/CR). Also <em>in vitro</em> – <em>in vivo</em> extrapolation modelling is available.</p> <div style="height:10px" aria-hidden="true" class="wp-block-spacer"></div> <hr class="wp-block-separator has-text-color has-symeres-font-color has-alpha-channel-opacity has-symeres-font-background-color has-background is-style-wide" /> <div style="height:10px" aria-hidden="true" class="wp-block-spacer"></div> <h2 class="wp-block-heading"><span class="ez-toc-section" id="Metabolic_Stability"></span>Metabolic Stability<span class="ez-toc-section-end"></span></h2> <ul class="wp-block-list"> <li class="has-medium-font-size"><strong><em>In vitro</em> metabolism in hepatocytes, liver microsomes, S9, recombinant enzymes or extrahepatic tissues, a co-culture model for low clearance compounds</strong></li> <li class="has-medium-font-size"><strong>Half-life, <em>in vitro</em> clearance</strong></li> <li class="has-medium-font-size"><strong><em>In vivo</em> extrapolation (to predict hepatic clearance & extraction ratio)</strong></li> </ul> <p class="has-medium-font-size">Measuring the metabolic stability of a compound <em>in vitro</em> provides an estimate regarding its stability, and thus elimination rate by metabolism in the body. Hepatic clearance is the most important parameter when assessing metabolic stability, and the most common approaches for its prediction are <em>in vitro</em> assays using liver microsomes or hepatocytes; the former focusing mainly on oxidative (CYP-mediated) metabolism, and the latter also on conjugative metabolism. For low clearance compounds, co-culture or sandwich models can be applied for investigating metabolic stability over several days.</p> <p class="has-medium-font-size">When measuring the disappearance rate of a compound under linear velocity conditions <em>in vitro</em>, the results can be further extrapolated to <em>in vivo</em> hepatic clearance and extraction ratio, and also to evaluate the effect of hepatic first-pass metabolism to total oral bioavailability. When using UPLC/high-resolution-MS for sample analysis, also metabolite identification can be easily delivered from the same analytical data.</p> <p></p> <div class="wp-block-buttons is-layout-flex wp-block-buttons-is-layout-flex"> <div class="wp-block-button has-custom-font-size is-style-outline has-gibson-font-family is-style-outline--8" style="font-size:18px;font-style:normal;font-weight:500;text-decoration:none;text-transform:none"><a class="wp-block-button__link has-symeres-white-color has-symeres-secondary-background-color has-text-color has-background wp-element-button" href="https://admeshop.com/collections/in-vitro-metabolism" style="border-radius:100px" target="_blank" rel="noopener"><strong>Request a quote via our Admeshop</strong></a></div> </div> <p></p> <section class="downloads downloadable-content-block"> <div class="downloads__list"> <form class="downloads"> <ul> <li> <input id="download-file-Admescope-Metabolic-stability-factsheet.pdf" name="download" data-filetitle="Admescope Metabolic stability factsheet" data-filename="Admescope-Metabolic-stability-factsheet.pdf" data-download="4864" name="download-file" type="checkbox"> <label for="download-file-Admescope-Metabolic-stability-factsheet.pdf"> Admescope Metabolic stability factsheet <span> <svg class="downloads-unchecked"><use xlink:href="#downloads-checkbox-default"></use></svg> <svg class="downloads-checked"><use xlink:href="#downloads-checkbox-checked"></use></svg> </span> </label> </li> <li> <input id="download-file-Admescope-Low-clearance-metabolism-factsheet.pdf" name="download" data-filetitle="Admescope Low clearance metabolism factsheet" data-filename="Admescope-Low-clearance-metabolism-factsheet.pdf" data-download="4863" name="download-file" type="checkbox"> <label for="download-file-Admescope-Low-clearance-metabolism-factsheet.pdf"> Admescope Low clearance metabolism factsheet <span> <svg class="downloads-unchecked"><use xlink:href="#downloads-checkbox-default"></use></svg> <svg class="downloads-checked"><use xlink:href="#downloads-checkbox-checked"></use></svg> </span> </label> </li> </ul> </form> </div> </section> <h2 class="wp-block-heading"><span class="ez-toc-section" id="Metabolite_Identification_and_Profiling"></span>Metabolite Identification and Profiling<span class="ez-toc-section-end"></span></h2> <figure class="wp-block-image alignright size-full"><img fetchpriority="high" decoding="async" width="336" height="386" src="https://www.admescope.com/wp-content/uploads/sites/2/2023/05/in-vitro-metabolism-1.png" alt="Admescope services for in vitro Metabolism research" class="wp-image-847" srcset="https://admescope.com/wp-content/uploads/sites/2/2023/05/in-vitro-metabolism-1.png 336w, https://admescope.com/wp-content/uploads/sites/2/2023/05/in-vitro-metabolism-1-261x300.png 261w" sizes="(max-width: 336px) 100vw, 336px" /></figure> <ul class="wp-block-list"> <li class="has-medium-font-size"><strong><em>In vitro</em> metabolism in hepatocytes, liver microsomes, S9, recombinant enzymes or extrahepatic tissues</strong></li> <li class="has-medium-font-size"><strong>Small drug-like compounds, peptides and antibody-drug conjugates</strong></li> <li class="has-medium-font-size"><strong>Metabolic soft-spots</strong></li> <li class="has-medium-font-size"><strong>Cross-species comparison</strong></li> <li class="has-medium-font-size"><strong>UPLC with Q-TOF-MS or QE-Orbitrap-MS and UV detection</strong></li> <li class="has-medium-font-size"><strong>UPLC with on-line or off-line radiodetection</strong></li> <li class="has-medium-font-size"><strong>Isolation of metabolites</strong></li> </ul> <p class="has-medium-font-size">Metabolite identification and profiling are relevant in several stages of drug discovery & development. Optimization of metabolic clearance requires information on the main metabolically labile “soft-spots” and their further chemical modification. Selection of suitable animal species for toxicity studies is facilitated by the prediction of species with most similar metabolite profiles with respect to human (first <em>in vitro</em> and then <em>in vivo</em>). The correct evaluation of metabolic enzymes involved in clearance also requires identification of the metabolites, and eventually the metabolic clearance routes in human <em>in vivo</em> has to be characterized in a detailed level.</p> <p class="has-medium-font-size">For <em>in vitro</em> this typically requires studies with enzyme sources such as liver (or extrahepatic) microsomes, S9 fraction, hepatocytes, and recombinant enzymes, while for <em>in vivo</em> the plasma and excreta are the major sample types. Analytically the UPLC/high-resolution-MS/MS techniques are the major tool for metabolite identification in all study types, supported partially by additional detection technologies such as UV/PDA, or radiodetection for the studies where 3H or 14C-labeled compounds are used. Upscaled production and isolation of the metabolite is also a valuable tool for both qualitative and quantitative purposes. For peptides, the metabolism (catabolism) is most often based on amide/peptide-bond hydrolysis reactions, which are efficiently identified by UPLC/high-resolution MS techniques. Besides small molecules and peptides, we offer metabolite <a href="https://admescope.com/services/biologics/#ADC_Characterization">identification services for antibody-drug conjugates (ADCs)</a> as part of our portfolio for biologics.</p> <p></p> <div class="wp-block-buttons is-layout-flex wp-block-buttons-is-layout-flex"> <div class="wp-block-button has-custom-font-size is-style-outline has-gibson-font-family is-style-outline--9" style="font-size:18px;font-style:normal;font-weight:500;text-decoration:none;text-transform:none"><a class="wp-block-button__link has-symeres-white-color has-symeres-secondary-background-color has-text-color has-background wp-element-button" href="https://admeshop.com/collections/in-vitro-metabolism" style="border-radius:100px" target="_blank" rel="noopener"><strong>Request a quote via our Admeshop</strong></a></div> </div> <p></p> <section class="downloads downloadable-content-block"> <div class="downloads__list"> <form class="downloads"> <ul> <li> <input id="download-file-Admescope-Metabolite-identification-factsheet.pdf" name="download" data-filetitle="Admescope Metabolite identification factsheet" data-filename="Admescope-Metabolite-identification-factsheet.pdf" data-download="4865" name="download-file" type="checkbox"> <label for="download-file-Admescope-Metabolite-identification-factsheet.pdf"> Admescope Metabolite identification factsheet <span> <svg class="downloads-unchecked"><use xlink:href="#downloads-checkbox-default"></use></svg> <svg class="downloads-checked"><use xlink:href="#downloads-checkbox-checked"></use></svg> </span> </label> </li> <li> <input id="download-file-Admescope-Peptide-metabolite-identification-factsheet.pdf" name="download" data-filetitle="Admescope Peptide metabolite identification factsheet" data-filename="Admescope-Peptide-metabolite-identification-factsheet.pdf" data-download="4866" name="download-file" type="checkbox"> <label for="download-file-Admescope-Peptide-metabolite-identification-factsheet.pdf"> Admescope Peptide metabolite identification factsheet <span> <svg class="downloads-unchecked"><use xlink:href="#downloads-checkbox-default"></use></svg> <svg class="downloads-checked"><use xlink:href="#downloads-checkbox-checked"></use></svg> </span> </label> </li> </ul> </form> </div> </section> <h2 class="wp-block-heading"><span class="ez-toc-section" id="Phenotyping_of_Metabolizing_Enzymes"></span>Phenotyping of Metabolizing Enzymes<span class="ez-toc-section-end"></span></h2> <ul class="wp-block-list"> <li class="has-medium-font-size"><strong>Liver microsomes and CYP-selective inhibitors</strong></li> <li class="has-medium-font-size"><strong>Recombinant CYP, FMO, MAO, CE, AOX, UGT, SULT and NAT</strong></li> <li class="has-medium-font-size"><strong>UPLC with Q-TOF-MS or QE-Orbitrap-MS</strong></li> </ul> <figure class="wp-block-image alignleft size-full"><img decoding="async" width="336" height="386" src="https://www.admescope.com/wp-content/uploads/sites/2/2023/05/In-vitro-Toxicology-1.png" alt="Work with phenotyping of Metabolizing Enzymes" class="wp-image-959" srcset="https://admescope.com/wp-content/uploads/sites/2/2023/05/In-vitro-Toxicology-1.png 336w, https://admescope.com/wp-content/uploads/sites/2/2023/05/In-vitro-Toxicology-1-261x300.png 261w" sizes="(max-width: 336px) 100vw, 336px" /></figure> <p class="has-medium-font-size">Enzyme phenotyping provides information on the enzymes involved in the metabolism of a new chemical entity, which is needed to evaluate the drug interaction vulnerability of the compound as a “victim-drug”. The phenotyping of metabolizing enzymes is also needed to evaluate the risk for highly variable clearance between individuals or populations due to the involvement of enzymes with high degree of polymorphism. A thorough evaluation of metabolizing enzymes also requires metabolite identification.</p> <p class="has-medium-font-size">The most common approach for enzyme identification is the use of recombinant enzymes and extrapolation of the results based on the <em>in vivo</em> protein levels, and the use of liver microsomal incubations together with cytochrome P450 (CYP)-selective inhibitors under linear metabolite formation kinetics. In both methods, monitoring of metabolite formation is clearly more reliable than monitoring substrate depletion only, although the latter is often used for high throughput evaluation. The clever use of HR-MS technology allows metabolite monitoring later on from the very same LC/MS data, even if only substrate depletion was analyzed in the first place. For more thorough evaluation of <em>in vivo</em> role of each enzyme, evaluation of kinetic parameters Vmax and Km with different enzymes would be advisable.</p> <p></p> <div class="wp-block-buttons is-layout-flex wp-block-buttons-is-layout-flex"> <div class="wp-block-button has-custom-font-size is-style-outline has-gibson-font-family is-style-outline--10" style="font-size:18px;font-style:normal;font-weight:500;text-decoration:none;text-transform:none"><a class="wp-block-button__link has-symeres-white-color has-symeres-secondary-background-color has-text-color has-background wp-element-button" href="https://admeshop.com/collections/in-vitro-metabolism" style="border-radius:100px" target="_blank" rel="noopener"><strong>Request a quote via our Admeshop</strong></a></div> </div> <p></p> <section class="downloads downloadable-content-block"> <div class="downloads__list"> <form class="downloads"> <ul> <li> <input id="download-file-Admescope-CYP-phenotyping-factsheet.pdf" name="download" data-filetitle="Admescope CYP phenotyping factsheet" data-filename="Admescope-CYP-phenotyping-factsheet.pdf" data-download="4868" name="download-file" type="checkbox"> <label for="download-file-Admescope-CYP-phenotyping-factsheet.pdf"> Admescope CYP phenotyping factsheet <span> <svg class="downloads-unchecked"><use xlink:href="#downloads-checkbox-default"></use></svg> <svg class="downloads-checked"><use xlink:href="#downloads-checkbox-checked"></use></svg> </span> </label> </li> <li> <input id="download-file-Admescope-Low-clearance-CYP-phenotyping-factsheet.pdf" name="download" data-filetitle="Admescope Low clearance CYP phenotyping factsheet" data-filename="Admescope-Low-clearance-CYP-phenotyping-factsheet.pdf" data-download="4862" name="download-file" type="checkbox"> <label for="download-file-Admescope-Low-clearance-CYP-phenotyping-factsheet.pdf"> Admescope Low clearance CYP phenotyping factsheet <span> <svg class="downloads-unchecked"><use xlink:href="#downloads-checkbox-default"></use></svg> <svg class="downloads-checked"><use xlink:href="#downloads-checkbox-checked"></use></svg> </span> </label> </li> <li> <input id="download-file-Admescope-UGT-phenotyping-factsheet.pdf" name="download" data-filetitle="Admescope UGT phenotyping factsheet" data-filename="Admescope-UGT-phenotyping-factsheet.pdf" data-download="4869" name="download-file" type="checkbox"> <label for="download-file-Admescope-UGT-phenotyping-factsheet.pdf"> Admescope UGT phenotyping factsheet <span> <svg class="downloads-unchecked"><use xlink:href="#downloads-checkbox-default"></use></svg> <svg class="downloads-checked"><use xlink:href="#downloads-checkbox-checked"></use></svg> </span> </label> </li> </ul> </form> </div> </section> <h2 class="wp-block-heading"><span class="ez-toc-section" id="Non-CYP_Mediated_Metabolism"></span>Non-CYP Mediated Metabolism<span class="ez-toc-section-end"></span></h2> <ul class="wp-block-list"> <li class="has-medium-font-size"><strong>Metabolic stability and metabolite identification</strong></li> <li class="has-medium-font-size"><strong>Phenotyping of metabolizing enzymes</strong></li> <li class="has-medium-font-size"><strong>UGT, SULT, NAT, FMO, MAO, CE, ALDH, AOX, XO, AKR, CR</strong></li> <li class="has-medium-font-size"><strong>Recombinant enzymes, liver microsomes, S9, cytosol, and hepatocytes or extrahepatic tissues</strong></li> </ul> <p class="has-medium-font-size">If CYP-focused enzyme phenotyping assays do not show clear results, non-CYP mediated metabolism should be studied and the enzymes involved in the metabolic processes identified. This can be done by using recombinant enzymes, such as UDP glucuronosyltransferases (UGT), sulfotransferases (SULT), N-acetyltransferases (NAT), flavin containing mono-oxygenases (FMO), monoamine-oxidases (MAO), carboxylesterases (CE), or aldehyde dehydrogenase (ALDH). Metabolism via aldehyde oxidase (AOX), xanthine oxidase (XO), aldo-keto reductase (AKR) and carbonyl reductase (CR) can be investigated by using human liver cytosol with enzyme-selective inhibitors.</p> <p></p> <div class="wp-block-buttons is-layout-flex wp-block-buttons-is-layout-flex"> <div class="wp-block-button has-custom-font-size is-style-outline has-gibson-font-family is-style-outline--11" style="font-size:18px;font-style:normal;font-weight:500;text-decoration:none;text-transform:none"><a class="wp-block-button__link has-symeres-white-color has-symeres-secondary-background-color has-text-color has-background wp-element-button" href="https://admeshop.com/collections/in-vitro-metabolism" style="border-radius:100px" target="_blank" rel="noopener"><strong>Request a quote via our Admeshop</strong></a></div> </div> <p></p> <div style="height:10px" aria-hidden="true" class="wp-block-spacer"></div> <hr class="wp-block-separator has-text-color has-symeres-font-color has-alpha-channel-opacity has-symeres-font-background-color has-background is-style-wide" /> <div style="height:10px" aria-hidden="true" class="wp-block-spacer"></div> <h2 class="wp-block-heading"><span class="ez-toc-section" id="Extrahepatic_Metabolism"></span>Extrahepatic Metabolism<span class="ez-toc-section-end"></span></h2> <figure class="wp-block-image alignright size-full"><img decoding="async" width="336" height="386" src="https://www.admescope.com/wp-content/uploads/sites/2/2023/05/in-vitro-metabolism-3.png" alt="Admescope lab team member working with Extrahepatic Metabolism" class="wp-image-857" srcset="https://admescope.com/wp-content/uploads/sites/2/2023/05/in-vitro-metabolism-3.png 336w, https://admescope.com/wp-content/uploads/sites/2/2023/05/in-vitro-metabolism-3-261x300.png 261w" sizes="(max-width: 336px) 100vw, 336px" /></figure> <ul class="wp-block-list"> <li class="has-medium-font-size"><strong><em>In vitro</em> metabolism in intestine, kidney, skin, plasma</strong></li> <li class="has-medium-font-size"><strong>Microsomes, S9 and homogenates</strong></li> </ul> <p class="has-medium-font-size">Although the liver is the major drug metabolizing organ in the body, other tissues, such as intestine, kidney and skin may also play a significant role in the metabolism of xenobiotic compounds. The role of each organ varies depending on the administration route and distribution of the compound between the organs. The abundance of metabolizing enzymes (CYPs and conjugative enzymes) also varies in each organ/tissue even with qualitative differences. From the experimental point of view, different tissue subfractions are routinely used together with modern analytical techniques. For more thorough evaluation of <em>in vivo</em> role of each organ, evaluation of kinetic parameters Vmax and Km with organ-dependent preparates would be advisable.</p> <p></p> <div class="wp-block-buttons is-content-justification-left is-layout-flex wp-container-core-buttons-is-layout-12 wp-block-buttons-is-layout-flex"> <div class="wp-block-button has-custom-font-size is-style-outline has-gibson-font-family is-style-outline--12" style="font-size:18px;font-style:normal;font-weight:500;text-decoration:none;text-transform:none"><a class="wp-block-button__link has-symeres-white-color has-symeres-secondary-background-color has-text-color has-background wp-element-button" href="https://admeshop.com/collections/in-vitro-metabolism" style="border-radius:100px" target="_blank" rel="noopener"><strong>Request a quote via our Admeshop</strong></a></div> </div> <p></p> <div style="height:10px" aria-hidden="true" class="wp-block-spacer"></div> <hr class="wp-block-separator has-text-color has-symeres-font-color has-alpha-channel-opacity has-symeres-font-background-color has-background is-style-wide" /> <div style="height:10px" aria-hidden="true" class="wp-block-spacer"></div> <h2 class="wp-block-heading"><span class="ez-toc-section" id="Metabolite_Production"></span>Metabolite Production<span class="ez-toc-section-end"></span></h2> <ul class="wp-block-list"> <li class="has-medium-font-size"><strong>Production using recombinant enzymes, liver microsomes or biomimetic catalysts</strong></li> </ul> <p class="has-medium-font-size">When confirmation of structure of the drug metabolite or its quantification is needed, we have a selection of biological and biomimetic tools available to upscale the metabolite production, followed by isolation using preparative chromatography.</p> <div style="height:10px" aria-hidden="true" class="wp-block-spacer"></div> <hr class="wp-block-separator has-text-color has-symeres-font-color has-alpha-channel-opacity has-symeres-font-background-color has-background is-style-wide" /> <div style="height:10px" aria-hidden="true" class="wp-block-spacer"></div> <h2 class="wp-block-heading"><span class="ez-toc-section" id="Reactive_Metabolites"></span>Reactive Metabolites<span class="ez-toc-section-end"></span></h2> <figure class="wp-block-image alignleft size-full"><img loading="lazy" decoding="async" width="337" height="386" src="https://www.admescope.com/wp-content/uploads/sites/2/2023/05/in-vitro-metabolism-4.png" alt="Admescope lab team members working with Reactive Metabolites" class="wp-image-860" srcset="https://admescope.com/wp-content/uploads/sites/2/2023/05/in-vitro-metabolism-4.png 337w, https://admescope.com/wp-content/uploads/sites/2/2023/05/in-vitro-metabolism-4-262x300.png 262w" sizes="auto, (max-width: 337px) 100vw, 337px" /></figure> <ul class="wp-block-list"> <li class="has-medium-font-size"><strong>GSH, KCN, semicarbazide trapping with</strong> <strong>UPLC/HR-MS detection</strong></li> <li class="has-medium-font-size"><strong>Covalent binding with radiolabeled compounds</strong></li> </ul> <p class="has-medium-font-size">One of the reasons behind unexpected drug induced toxicity can be chemically reactive metabolites that bind covalently to protein or DNA. Due to their high reactivity and short life<del>–</del>span, these metabolites cannot be analyzed directly as such, but have to be first stabilized by trapping agents, such as S-glutathione (GSH), cyanide (KCN) or semicarbazide (SCA). Of these, GSH is clearly the most used trapping agent, although KCN and SCA may be used for trapping certain types of reactive metabolites that are not trapped by GSH, such as iminium ions or aldehydes. As in any analysis of unknown drug metabolites, the use of liquid chromatography with high-resolution mass spectrometry delivers the highest quality data with the shortest time consumption also in our reactive metabolite screening assay.</p> <p class="has-medium-font-size">In addition to screening, we can also perform a more comprehensive quantitative evaluation of reactive metabolite formation by utilizing a radiolabeled compound and measuring the radioactivity of the reactive metabolites covalently bound to proteins within the <em>in vitro</em> metabolic system. </p> <p></p> <div class="wp-block-buttons is-layout-flex wp-block-buttons-is-layout-flex"> <div class="wp-block-button has-custom-font-size is-style-outline has-gibson-font-family is-style-outline--13" style="font-size:18px;font-style:normal;font-weight:500;text-decoration:none;text-transform:none"><a class="wp-block-button__link has-symeres-white-color has-symeres-secondary-background-color has-text-color has-background wp-element-button" href="https://admeshop.com/collections/in-vitro-metabolism" style="border-radius:100px" target="_blank" rel="noopener"><strong>Request a quote via our Admeshop</strong></a></div> </div> <p></p> <section class="downloads downloadable-content-block"> <div class="downloads__list"> <form class="downloads"> <ul> <li> <input id="download-file-Admescope-Reactive-metabolites-factsheet.pdf" name="download" data-filetitle="Admescope Reactive metabolites factsheet" data-filename="Admescope-Reactive-metabolites-factsheet.pdf" data-download="4875" name="download-file" type="checkbox"> <label for="download-file-Admescope-Reactive-metabolites-factsheet.pdf"> Admescope Reactive metabolites factsheet <span> <svg class="downloads-unchecked"><use xlink:href="#downloads-checkbox-default"></use></svg> <svg class="downloads-checked"><use xlink:href="#downloads-checkbox-checked"></use></svg> </span> </label> </li> <li> <input id="download-file-Admescope-Covalent-binding-factsheet.pdf" name="download" data-filetitle="Admescope Covalent binding factsheet" data-filename="Admescope-Covalent-binding-factsheet.pdf" data-download="4874" name="download-file" type="checkbox"> <label for="download-file-Admescope-Covalent-binding-factsheet.pdf"> Admescope Covalent binding factsheet <span> <svg class="downloads-unchecked"><use xlink:href="#downloads-checkbox-default"></use></svg> <svg class="downloads-checked"><use xlink:href="#downloads-checkbox-checked"></use></svg> </span> </label> </li> </ul> </form> </div> </section> <h2 class="wp-block-heading"><span class="ez-toc-section" id="Acyl_Glucuronide_Reactivity"></span>Acyl Glucuronide Reactivity<span class="ez-toc-section-end"></span></h2> <ul class="wp-block-list"> <li class="has-medium-font-size"><strong>Stability and reactivity of acyl glucuronide metabolites</strong></li> <li class="has-medium-font-size"><strong>No need for synthesized metabolite</strong></li> <li class="has-medium-font-size"><strong>UPLC/HR-MS detection</strong></li> </ul> <p class="has-medium-font-size">Although glucuronide conjugation is most often considered as a detoxifying metabolic route, acyl glucuronide conjugates, formed in the presence of carboxylic acid group, may cause toxicity by covalent binding to proteins. Liability for toxicity of an acyl glucuronide metabolite may be evaluated by measuring the rate of acyl-migration, i.e., formation of isomeric acyl glucuronides and eventually formation of reactive aldehyde isomer. This method is a valuable tool for assessing acyl glucuronide reactivity without the need for synthetized glucuronide metabolites. In addition to acyl glucuronide conjugates, the formation of even more reactive Acyl CoA conjugates or other related conjugates (glycine, taurine, carnitine) may be investigated by our methods.</p> <p></p> <div class="wp-block-buttons is-layout-flex wp-block-buttons-is-layout-flex"> <div class="wp-block-button has-custom-font-size is-style-outline has-gibson-font-family is-style-outline--14" style="font-size:18px;font-style:normal;font-weight:500;text-decoration:none;text-transform:none"><a class="wp-block-button__link has-symeres-white-color has-symeres-secondary-background-color has-text-color has-background wp-element-button" href="https://admeshop.com/collections/in-vitro-metabolism" style="border-radius:100px" target="_blank" rel="noopener"><strong>Request a quote via our Admeshop</strong></a></div> </div> <p></p> <section class="downloads downloadable-content-block"> <div class="downloads__list"> <form class="downloads"> <ul> <li> <input id="download-file-Admescope-Acyl-glucuronide-reactivity-factsheet.pdf" name="download" data-filetitle="Admescope Acyl glucuronide reactivity factsheet" data-filename="Admescope-Acyl-glucuronide-reactivity-factsheet.pdf" data-download="4873" name="download-file" type="checkbox"> <label for="download-file-Admescope-Acyl-glucuronide-reactivity-factsheet.pdf"> Admescope Acyl glucuronide reactivity factsheet <span> <svg class="downloads-unchecked"><use xlink:href="#downloads-checkbox-default"></use></svg> <svg class="downloads-checked"><use xlink:href="#downloads-checkbox-checked"></use></svg> </span> </label> </li> </ul> </form> </div> </section> <p></p> </div></div> <section class="block block--primary"> <div class="block__content block__content--right"> <div class="block__text"> <h2>Feel free to contact us!</h2> <p><strong>Curious to know more about what can we do for you? 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name="email" type="email" placeholder="Email" required> </div> </div> <div class="form__row"> <div class="form__col"> <label for="agreement"> <input id="agreement" name="agreement" type="checkbox" required> <div>By downloading, I agree to the <a href='/privacy-policy/'>privacy policy</a></div> </label> </div> </div> <div class="form__row"> <div class="form__col"> <input type="text" name="downloaded_files" value="" style="opacity: 0;height: 0;overflow: hidden;"> <button type="submit" class="button button--primary">Download file(s)</button> <input type="hidden" name="action" value="symeresdownloads"> </div> </div> </form> </div> </section><script> jQuery( document ).ready(function() { jQuery(function ($) { let downloads = {}; $('body').on('click', '#downloads-close', function(){ $('.downloads__label').css('display', 'flex'); $('.downloads__content').css('display', 'none'); }); $('body').on('click', '.downloads__label', function(){ $('.downloads__label').css('display', 'none'); $('.downloads__content').css('display', 'flex'); }); $('body').on('change', 'form.downloads', function(){ let downloadsList = ''; let filenames = {}; $('input:checkbox[name=download]').each(function(){ filenames[$(this).data('filename')] = $(this).data('filetitle'); if (this.checked) { downloads[$(this).data('download')] = $(this).data('filename'); } else { if ($(this).data('download') in downloads) { delete downloads[$(this).data('download')]; } if (localStorage.getItem('downloads')) { let downloadsFromLocalStorage = JSON.parse(localStorage.getItem('downloads')); if ($(this).data('download') in downloadsFromLocalStorage) { delete downloadsFromLocalStorage[$(this).data('download')]; localStorage.setItem('downloads', JSON.stringify(downloadsFromLocalStorage)); } } } $('input[name="downloaded_files"]').val(Object.values(filenames).join(',')); }); // set filenames + titles in localStorage if (Object.keys(filenames).length) { let filenamesJson = JSON.stringify(filenames); if (localStorage.getItem('filenames')) { let currentFilenames = JSON.parse(localStorage.getItem('filenames')); filenames = { ...currentFilenames, ...filenames }; filenamesJson = JSON.stringify(filenames); } localStorage.setItem('filenames', filenamesJson); } if (Object.keys(downloads).length) { $('.downloads__aside').css('display', 'block'); let downloadsJson = JSON.stringify(downloads); if (localStorage.getItem('downloads')) { let currentDownloads = JSON.parse(localStorage.getItem('downloads')); downloads = { ...currentDownloads, ...downloads }; downloadsJson = JSON.stringify(downloads); } localStorage.setItem('downloads', downloadsJson); } else { if (!localStorage.getItem('downloads')) { localStorage.removeItem('downloads'); } } Object.keys(downloads).forEach(key => { let label = $('.downloads li label[for="download-file-' + downloads[key] + '"]').text(); if (localStorage.getItem('downloads')) { let downloadsFromLocalStorage = JSON.parse(localStorage.getItem('downloads')); if (localStorage.getItem('filenames')) { let filenames = JSON.parse(localStorage.getItem('filenames')); if (downloadsFromLocalStorage[key] in filenames) { label = filenames[downloadsFromLocalStorage[key]]; } } if (label === '') { label = $('.downloads li label[for="download-file-' + downloadsFromLocalStorage[key] + '"]').text(); } } downloadsList += '<li><button data-download="' + key + '"><svg class="downloads__delete"><use xlink:href="#icon-times"></use></svg></button><span><svg class="downloads__icon"><use xlink:href="#icon-download-file"></use></svg></span>' + label.trim() + '</li>'; }); $('.downloads__label span').text(Object.keys(downloads).length); $('#selected-files ul').html(downloadsList); // if (Object.keys(downloads).length) { //if (localStorage.getItem('downloads')) { // $('.downloads__aside').css('display', 'block'); // } else { // $('.downloads__aside').css('display', 'none'); // } }); if (localStorage.getItem('downloads')) { let downloadsFromLocalStorage = JSON.parse(localStorage.getItem('downloads')); let downloadsList = ''; $("input[name='downloaded_files']").val(Object.values(downloadsFromLocalStorage).join(',')); Object.keys(downloadsFromLocalStorage).forEach(key => { $(".downloads ul").find("[data-filename='" + downloadsFromLocalStorage[key] + "']").prop('checked', true); let label = ''; if (localStorage.getItem('filenames')) { let filenames = JSON.parse(localStorage.getItem('filenames')); if (downloadsFromLocalStorage[key] in filenames) { label = filenames[downloadsFromLocalStorage[key]]; } } if (label === '') { label = $('.downloads li label[for="download-file-' + downloadsFromLocalStorage[key] + '"]').text(); } downloadsList += '<li><button data-download="' + key + '"><svg class="downloads__delete"><use xlink:href="#icon-times"></use></svg></button><span><svg class="downloads__icon"><use xlink:href="#icon-download-file"></use></svg></span>' + label.trim() + '</li>'; }); $('.downloads__label span').text(Object.keys(downloadsFromLocalStorage).length); $('#selected-files ul').html(downloadsList); $('.downloads__aside').css('display', 'block'); } else { $('.downloads__aside').css('display', 'none'); } $('body').on('click', '#remove-all-files', function(){ localStorage.removeItem('downloads'); $('#selected-files div ul li').remove(); $('.downloads__aside').css('display', 'none'); location.reload(); }); $('body').on('click', '#selected-files button', function(){ $('form.downloads input[data-download=' + $(this).data('download') + ']').prop('checked', false).trigger('change'); if (localStorage.getItem('downloads')) { let downloadsLocalObj = JSON.parse(localStorage.getItem('downloads')); if ($(this).data('download') in downloadsLocalObj) { delete downloadsLocalObj[$(this).data('download')]; localStorage.setItem('downloads', JSON.stringify(downloadsLocalObj)); //reload sidebar html let downloadsList = ''; Object.keys(downloadsLocalObj).forEach(key => { $(".downloads ul").find("[data-filename='" + downloadsLocalObj[key] + "']").prop('checked', true); let label = ''; if (localStorage.getItem('filenames')) { let filenames = JSON.parse(localStorage.getItem('filenames')); if (downloadsLocalObj[key] in filenames) { label = filenames[downloadsLocalObj[key]]; } } if (label === '') { label = $('.downloads li label[for="download-file-' + downloadsLocalObj[key] + '"]').text(); } downloadsList += '<li><button data-download="' + key + '"><svg class="downloads__delete"><use xlink:href="#icon-times"></use></svg></button><span><svg class="downloads__icon"><use xlink:href="#icon-download-file"></use></svg></span>' + label.trim() + '</li>'; }); $('.downloads__label span').text(Object.keys(downloadsLocalObj).length); $('#selected-files ul').html(downloadsList); $('.downloads__aside').css('display', 'block'); } } }); $('#symeresdownloads').on('submit', function (e) { e.preventDefault(); if (localStorage.getItem('downloads')) { downloads = JSON.parse(localStorage.getItem('downloads')); $("input[name='downloaded_files']").val(''); $("input[name='downloaded_files']").val(Object.values(downloads).join(',')); } var symeresdownloads = $('#symeresdownloads'); var email = $("#symeresdownloads :input[name='email']").val(); var firstName = $("#symeresdownloads :input[name='first-name']").val(); var lastName = $("#symeresdownloads :input[name='last-name']").val(); var company = $("#symeresdownloads :input[name='company']").val(); var action = $("#symeresdownloads :input[name='action']").val(); downloads['email'] = email; downloads['first-name'] = firstName; downloads['last-name'] = lastName; downloads['company'] = company; downloads['action'] = action; runAjaxCall(symeresdownloads, downloads); // Clean up the downloads object delete downloads['email']; delete downloads['first-name']; delete downloads['last-name']; delete downloads['company']; delete downloads['action']; e.preventDefault(); }); function runAjaxCall(symeresdownloads, downloads) { var array = Object.keys(downloads).map((key) => [String(key), downloads[key]]); $.ajax({ url: symeresdownloads.attr('action'), data: downloads, // form data type: symeresdownloads.attr('method'), // POST beforeSend: function (xhr) { jQuery('#symeresdownloads button').hide(); jQuery('<p class="download-status">Your file(s) are being processed.</p>').insertAfter('#symeresdownloads button'); }, success: function (data) { let error = false; if (data.includes('address') || data.includes('download')) { if (!data.includes('Thank you')) { error = true; jQuery('.download-status').remove(); jQuery('.downloads__content').append('<div class="status-msg" style="font-weight:500; color: #FFF;">' + data + '</div>') } } if (data.includes('Thank you')) { error = false; jQuery('.download-status').remove(); jQuery('.downloads__content').append('<div class="status-msg" style="font-weight:500; color: #FFF;">' + data + '</div>') } setTimeout(function(){ jQuery('.status-msg').remove(); if (error) { jQuery('input#email').css('border-color', '#d63637'); location.reload(); } if (!error) { localStorage.removeItem('downloads'); $('.downloads__aside').css('display', 'none'); location.reload(); } }, 5000); } }); } if ($('.downloads__label span').text() == '0') { $('.downloads__aside').css('display', 'none'); } $('body').on('change', '.downloads__label span', function(){ if ($('.downloads__label span').text() == '0') { $('.downloads__aside').css('display', 'none'); } }); }); }); </script><script id="wp-block-template-skip-link-js-after"> ( function() { var skipLinkTarget = document.querySelector( 'main' ), sibling, skipLinkTargetID, skipLink; // Early exit if a skip-link target can't be located. if ( ! skipLinkTarget ) { return; } /* * Get the site wrapper. * The skip-link will be injected in the beginning of it. */ sibling = document.querySelector( '.wp-site-blocks' ); // Early exit if the root element was not found. if ( ! sibling ) { 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