<h3 id="h-expedited-screening-improves-outcomes-for-individuals-with-drug-toxicity" data-uw-rm-heading="level" role="heading" aria-level="2"><strong>Expedited screening improves outcomes for individuals with drug toxicity</strong></h3><p><strong>In the field of clinical research and toxicology, the detection and quantification of drugs in human samples with suspected drug overdose or acute drug toxicity are critical. This blog post explores the potential of paper spray mass spectrometry (PS-MS) such as </strong><a target="_blank" rel="noreferrer noopener" aria-label="Thermo Scientific VeriSpray PaperSpray ion source - open in a new tab" data-uw-rm-ext-link="" uw-rm-external-link-id="https://www.thermofisher.com/us/en/home/industrial/mass-spectrometry/liquid-chromatography-mass-spectrometry-lc-ms/lc-ms-accessories/verispray-paperspray-ion-source-mass-spectrometry.html$thermoscientificverispraypapersprayionsource"><a target="_blank" rel="noopener noreferrer" href="https://www.thermofisher.com/us/en/home/industrial/mass-spectrometry/liquid-chromatography-mass-spectrometry-lc-ms/lc-ms-accessories/verispray-paperspray-ion-source-mass-spectrometry.html"><strong>Thermo Scientific VeriSpray PaperSpray ion source</strong></a></a><strong>, for faster and more efficient drug toxicity screening in emergency room applications.</strong></p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Thermo_Fisher_Scientific_Collaboration_Advances_POC_Solutions_Using_Paper_Spray_Mass_Spec_for_Drug_Tox_Screening_0531bbdb29.jpg" alt="Thermo Fisher Scientific: Collaboration Advances POC Solutions Using PaperSpray Mass Spec for Drug Tox Screening." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Thermo_Fisher_Scientific_Collaboration_Advances_POC_Solutions_Using_Paper_Spray_Mass_Spec_for_Drug_Tox_Screening_0531bbdb29.jpg 234w," sizes="100vw" width="1200" height="800"></p><article aria-label="Collaboration Advances POC Solutions Using PaperSpray Mass Spec for Drug Tox Screening"><div><p>Researchers from <a target="_blank" rel="noreferrer noopener" aria-label="King’s College London - open in a new tab" data-uw-rm-ext-link="" uw-rm-external-link-id="https://www.kcl.ac.uk/forensics$king’scollegelondon"><a target="_blank" rel="noopener noreferrer" href="https://www.kcl.ac.uk/forensics"><strong>King’s College London</strong></a></a> (KCL) are collaborating with <strong>Guy’s and St Thomas’ NHS Foundation Trust</strong> and leveraging the capabilities of <strong>paper spray-mass spectrometry (PS-MS) </strong>to analyze blood samples from individuals with acute drug toxicity in the emergency room. The aim is to <strong>develop routine methods for detecting and quantifying drugs</strong>, ultimately improving health outcomes for individuals in need of immediate care and making drug toxicity screening at the bedside a possibility in the future.</p><p>KCL researchers are conducting extensive testing and optimization of PS-MS technology. Preliminary results shared at <strong>The International Association of Forensic Toxicologists (TIAFT) 2023</strong> annual meeting in Rome have shown promising linearity across various concentrations for a number of cathinones. The goal is to refine the process and enhance accuracy, with the potential implementation of PS-MS into clinical practice.</p><h3 id="h-advantages-of-verispray-paperspray-mass-spectrometry"><strong>Advantages of VeriSpray PaperSpray mass spectrometry</strong></h3><p><strong>PS-MS with the</strong> <a target="_blank" rel="noreferrer noopener" aria-label="Thermo Scientific™ TSQ Altis™ Plus Triple Quadrupole - open in a new tab" data-uw-rm-ext-link="" uw-rm-external-link-id="https://www.thermofisher.com/order/catalog/product/tsq03-10002$thermoscientific™tsqaltis™plustriplequadrupole"><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/404"><strong>Thermo Scientific™ TSQ Altis™ Plus Triple Quadrupole</strong></a></a> mass spectrometer offers several advantages over traditional orthogonal techniques. It enables rapid analysis, eliminating the need for lengthy sample preparation and chromatography-based separation. Additionally, the technique provides quantitative or screening results, is easy to use, reduces the risk of cross-contamination, and has barcoded tracking for LIMS compatibility with robotic automated systems such as Hamilton. Plus, it minimizes the use of chemicals and solvents, making it more environmentally friendly.</p><p>PS-MS offers a fast (2 minutes or less) turnaround time for high-throughput screening, enabling quick analysis of single samples from individuals and identify the presence of drugs or substances. The system’s ease of use and ability to process 240 samples unattended make it efficient and time-saving. The aim is to make drug toxicity screening at the bedside a reality, reducing waiting times for results and improving overall care.</p><h3 id="h-great-potential-for-poc-toxicity-screening"><strong>Great potential for POC toxicity screening</strong></h3><p>The collaboration between KCL, Guy’s and St Thomas’ NHS Foundation Trust, and research on PS-MS holds great potential for improving point-of-care drug toxicity screening. PS-MS offers a rapid, efficient, and accurate alternative to traditional methods, enabling faster and more efficient care for individuals presenting with acute drug toxicity. By implementing this technology, hospitals can reduce waiting times and ultimately improve outcomes in emergency settings. To learn more, <a target="_blank" rel="noopener noreferrer" href="https://www.linkedin.com/posts/ema-ru%C5%BEi%C4%87-5ba70b133_paper-spray-mass-spectrometry-simplifies-activity-7196531937463750656-0FgR?utm_source=share&amp;utm_medium=member_desktop">download the recent case study</a>.</p><blockquote><h3 id="h-related-information"><strong>Related information</strong></h3><ol><li><a target="_blank" rel="noreferrer noopener" aria-label="Thermofisher.com/verispray - open in a new tab" data-uw-rm-ext-link="" uw-rm-external-link-id="https://www.thermofisher.com/us/en/home/industrial/mass-spectrometry/liquid-chromatography-mass-spectrometry-lc-ms/lc-ms-accessories/verispray-paperspray-ion-source-mass-spectrometry.html$thermofisher.com/verispray"><a target="_blank" rel="noopener noreferrer" href="https://www.thermofisher.com/us/en/home/industrial/mass-spectrometry/liquid-chromatography-mass-spectrometry-lc-ms/lc-ms-accessories/verispray-paperspray-ion-source-mass-spectrometry.html">Thermofisher.com/verispray</a></a></li><li><a target="_blank" rel="noreferrer noopener" aria-label="Toxicology in the emergency department: what’s new? - open in a new tab" data-uw-rm-ext-link="" uw-rm-external-link-id="https://pubmed.ncbi.nlm.nih.gov/36193928/#:~:text=intentional%20and%20accidental%20drug%20overdose,morbidity%20and%20prolonged%20hospital%20admissions.$toxicologyintheemergencydepartment:what’snew?"><a target="_blank" rel="noopener noreferrer" href="https://pubmed.ncbi.nlm.nih.gov/36193928/#:~:text=Intentional%20and%20accidental%20drug%20overdose,morbidity%20and%20prolonged%20hospital%20admissions.">Toxicology in the emergency department: what’s new?</a></a></li><li><a target="_blank" rel="noreferrer noopener" aria-label="Thermo Fisher Scientific collaboration offers potential for faster, more efficient analysis of samples from drug toxicity patients - open in a new tab" data-uw-rm-ext-link="" uw-rm-external-link-id="https://www.kcl.ac.uk/news/thermo-fisher-scientific-collaboration-offers-potential-for-faster-more-efficient-analysis-of-samples-from-drug-toxicity-patients$thermofisherscientificcollaborationofferspotentialforfaster,moreefficientanalysisofsamplesfromdrugtoxicitypatients"><a target="_blank" rel="noopener noreferrer" href="https://www.kcl.ac.uk/news/thermo-fisher-scientific-collaboration-offers-potential-for-faster-more-efficient-analysis-of-samples-from-drug-toxicity-patients">Thermo Fisher Scientific collaboration offers potential for faster, more efficient analysis of samples from drug toxicity patients</a></a></li><li><a target="_blank" rel="noreferrer noopener" aria-label="Paper spray mass spectrometry simplifies and expedites turnaround of drug toxicity screening: Thermo Fisher Scientific Case Study - open in a new tab" data-uw-rm-ext-link="" uw-rm-external-link-id="https://view.highspot.com/viewer/6644c7d165ad350931b0517c$paperspraymassspectrometrysimplifiesandexpeditesturnaroundofdrugtoxicityscreening:thermofisherscientificcasestudy"><a target="_blank" rel="noopener noreferrer" href="https://view.highspot.com/viewer/6644c7d165ad350931b0517c">Paper spray mass spectrometry simplifies and expedites turnaround of drug toxicity screening: Thermo Fisher Scientific Case Study</a></a></li></ol></blockquote></div></article><section><blockquote><h4>Courtney Patterson</h4><div itemprop="description">Courtney Patterson is an Application Scientist at Thermo Fisher Scientific, specializing in the development of LC-MS applications for forensic and clinical toxicology using targeted and untargeted approaches to quantify and confirm drugs of abuse.</div></blockquote></section>

Collaboration Advances POC Solutions Using PaperSpray Mass Spec for Drug Tox Screening

Hydrophilic interaction liquid chromatography: An efficient tool for assessing thorium interaction with phosphorylated biomimetic peptides

Quantitative Assessment of a Novel Device Designed for Patient-centric Sampling of Dried Plasma Using Targeted Proteomics

<p><strong>Introduced nearly 80 years ago, collision-based ion-activation and dissociation is the most commonly used type of fragmentation commercially available today. In principle, this technique is quite straightforward: apply energy to accelerate the particles of interest until they collide with an inert gas to induce fragmentation. Within these parameters, there are many different sub-categories of the technique. Here we will discuss three collision-based techniques that occur on Thermo Scientific mass spectrometers.</strong></p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Thermo_Fisher_Scientific_Freedom_to_Fragment_Introduction_The_importance_of_fragmentation_in_mass_spectrometry_1_c8f70d1281.jpg" alt="Thermo Fisher Scientific Freedom to Fragment Introduction" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Thermo_Fisher_Scientific_Freedom_to_Fragment_Introduction_The_importance_of_fragmentation_in_mass_spectrometry_1_c8f70d1281.jpg 245w," sizes="100vw" width="1024" height="512"></p><h3><strong>Higher-Energy Collisional Dissociation (HCD)</strong></h3><ol><li><strong>Process:</strong> HCD is a beam-based technique that occurs in the HCD cell or ion routing multipole, where the ion beam is directed into a potential well. Excited molecules collide with collision gas, causing fragmentation. This gas is typically nitrogen and is fast and efficient, but indiscriminate in technique. To ensure a higher energy collision, the potential energy is dropped (typically from the quadrupole into the collision cell) by many 10s of eV. This ensures the ions have high energy as they enter the cell with nitrogen and collide with enough energy for the fragmentation to occur. In peptides and proteins, HCD favors b- and y-ion formation. [Figure 1]</li><li><strong>Applications:</strong> HCD is commonly used for general proteomics and metabolomics.</li><li><strong>Considerations: </strong>When using Thermo Scientific mass spectrometers for HCD fragmentation, we generally use normalized collisional energy (NCE) instead of directly putting in the eV of potential well “drop”. The calibration routine determines the optimal voltage for a series of ions in the calibration solution.&nbsp; Then the routine scales these absolute voltages to a “normalized collision energy” value of 30. As such, an NCE value of 30 should work well across a range of <i>m/z</i> values and charge states. To see the actual applied value, one can check the scan header of the spectrum easily in our Thermo Scientific Freestyle software.</li></ol><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Thermo_Fisher_Scientific_Collision_Based_Ion_activation_and_Dissociation_92107a9524.jpg" alt="Thermo Fisher Scientific: Collision-Based Ion-activation and Dissociation: Figure 1: Location of peptide/protein backbone cleavage and its nomenclature. Representative here is a 4-amino acid peptide with side chains labeled R1-4." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Thermo_Fisher_Scientific_Collision_Based_Ion_activation_and_Dissociation_92107a9524.jpg 245w," sizes="100vw" width="1200" height="582"></p><h3><strong>Collision-Induced Dissociation (CID) (ion trap specific version)</strong></h3><ol><li><strong>Process: </strong>CID, as it occurs in Thermo Fisher Scientific’s dual linear ion trap technology, is a resonance-based technique where molecules are excited by a specific frequency.&nbsp; These excited molecules will start running into He molecules at a faster rate.&nbsp; These collisions slowly heat the resonant molecules until they fragment. Once fragmentation occurs, the product ions at new m/z values are no longer excited by the frequency and will cool back down to the center of the trap. In peptides and proteins, CID favors b- and y-ion formation.</li><li><strong>Applications: </strong>CID is great for small molecules and peptides and helps fragment oligonucleotides, especially when the q-value is adjusted.&nbsp; It also fragments with higher efficiency, which makes it well-suited for MS3 and higher workflows.</li><li><strong>Considerations: </strong>Because CID is resonance-based, there is a compromise outlined in the Matthieu equation. With certain voltages and frequencies applied, only certain m/z values can be trapped. Thus, we have something called the 1/3rd rule. This rule approximates the equation and states that any fragment created that is &lt; 1/3 the m/z of the precursor will not be trapped. This rule makes CID less desirable as a primary fragmentation technique, thus more used as the pinch hitter when needed for alternate fragmentation after HCD is shown inefficient for a specific molecule. Furthermore, CID is a slow heating process.&nbsp;As such, activation/fragmentation takes a few milliseconds, whereas HCD is effectively instantaneous.&nbsp;</li></ol><p>Recent developments in oligonucleotide analysis have shown great strides in using CID for fragmentation of many types, including tRNA, snRNA and digested mRNA. To gain the most impact from these experiments, scientists can adjust the q-value applied within the CID resonance. This is usually set to 0.25; however, for oligonucleotides, lowering that value to 0.2-0.15 can help prevent unwanted fragmentation from occurring and specify backbone cleavage that can be more easily characterized.</p><h3><strong>Source Induced Dissociation (SID)</strong></h3><ul><li><strong>Process: </strong>SID is a collisional-induced activation (and if desired, dissociation) technique that is induced in the source region of mass spectrometers. By applying more voltage in the source region, ions are drawn into the mass spectrometer with more energy. Within the source region, there are still atmospheric inert gases that have been drawn in with the analytes of interest. With the additional energy, collisions with these inert gases can occur. Since SID occurs before any mass analyzer, it applies to all ions that enter the mass spectrometer.</li><li><strong>Applications: </strong>SID is most commonly used as a desolvation technique for large native molecules. By applying SID energy in the source, molecules generate vibrational energy, and non-covalent, low-energy bonds will be broken only.</li><li><strong>Analogy: </strong>SID could be compared to shaking off an umbrella after coming in from the rain. The jostle doesn’t cause the whole umbrella to fall apart, but it does cause the water droplets to fall off. This is a little like proteins getting desolvated from SID.</li><li><strong>Considerations: </strong>SID can be a destructive, dissociative process as well, so if that is not the goal, it is important to monitor the energy applied to the analyte. For example, it has been common practice in top-down proteomics to use a small (10-15 eV) setting for SID to aid in desolvation. However, I have successfully taken pure protein (or well chromatographically separated protein) and applied much higher SID (&gt;50 V) to induce dissociation. The resulting fragmentation pattern can yield surprisingly good results. However, this is not common practice, especially not for coeluting species, as the fragmentation would occur on all proteins eluting.</li></ul><p>Another consideration for SID is its use in native MS. This is often vital for good detection of native proteins like antibodies. Often, we will employ a stronger SID in these cases, anywhere from 60-120 V, depending on the size and complexity of the protein/protein complex, to ensure proper desolvation and thus detection.</p><blockquote><p><strong>Learn more about </strong><a target="_blank" rel="noopener noreferrer" href="https://www.thermofisher.com/content/lifetech/global/en/home/industrial/mass-spectrometry/liquid-chromatography-mass-spectrometry-lc-ms/fragmentation-solutions.html"><strong>collisional induced dissociation (CID) and higher energy collisional induced dissociation (HCD)</strong></a></p><p><strong>Visit us on&nbsp;</strong><a target="_blank" rel="noopener noreferrer" href="https://www.linkedin.com/showcase/72003132/admin/dashboard/"><strong>LinkedIn:</strong></a>&nbsp;#CID #HCD #fragmentation</p></blockquote><section><blockquote><h4><strong>Amanda Lee</strong></h4><p>Amanda Lee PhD, Product Marketing Manager</p></blockquote></section>

Collision-Based Ion-activation and Dissociation

<p><strong>Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 15th September 2025? Check out new documents from the field of liquid phase, especially HPLC and LC/MS techniques!</strong></p><blockquote><p><span style="font-size:20px;">👉 </span><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library"><span style="font-size:20px;"><strong>SEARCH THE LARGEST REPOSITORY OF DOCUMENTS ABOUT LCMS AND RELATED TECHNIQUES</strong></span></a></p></blockquote><blockquote><p><strong>👉 Need info about different analytical techniques?</strong> Peek into <a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/library"><strong>LabRulezGCMS</strong></a> or <a target="_blank" rel="noopener noreferrer" href="https://icpms.labrulez.com/library"><strong>LabRulezICPMS</strong></a> libraries.</p></blockquote><p>This week we bring you application notes by <strong>Agilent Technologies, Shimadzu </strong>and <strong>Waters Corporation </strong>and<strong> </strong>poster by <strong>Thermo Fisher Scientific / HPLC Symposium!</strong></p><h3><strong>1. Agilent Technologies: A Direct Aqueous Injection Method for Contaminants in Drinking and Nonpotable Water</strong></h3><p><strong>Analysis of acrylamide, haloacetic acids, and β-estradiol in water with an Agilent 6495D LC/TQ</strong></p><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/an_aquesous_injection_method_water_6495d_lc_tq_5994_7973en_agilent_1_24e3a63a15.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Recent regulations, such as EU 2020/218, have included the addition of new emerging contaminants in drinking water and raw waters, which supply water-treatment works. Some of these new contaminants to be analyzed include acrylamide, HAAs, and β-estradiol. However, acrylamide and HAAs can be difficult to analyze due to their high polarity and high interferences from hard water samples. Additionally, β-estradiol requires a very low level of detection (1 ng/L) compared to the other compounds (acrylamide 0.1 µg/L and HAAs 60&nbsp;µg/L). The analysis uses a&nbsp;1290 Infinity II LC coupled to a&nbsp;6495D LC/TQ for analysis of water samples.</p><h4><strong>Experimental</strong></h4><h5><strong>Instrumentation&nbsp;</strong></h5><p>Chromatographic separation was performed using an <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=InfinityLab+Poroshell+120+Aq-C18"><strong>Agilent&nbsp;InfinityLab Poroshell 120 Aq-C18</strong></a>, 3.0 × 150&nbsp;mm, 2.7&nbsp;µm (part&nbsp;number 693675-742) installed on the&nbsp;1290&nbsp;Infinity II LC system. The individual modules of the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=1290+Infinity+II+LC"><strong>1290 Infinity II LC system</strong></a>&nbsp;included:</p><ul><li>Agilent 1290 Infinity II high‑speed pump (G7120A)&nbsp;</li><li>Agilent 1290 Infinity II autosampler (G7167B)&nbsp;</li><li>Agilent 1290 Infinity II multicolumn thermostat (G7116B)&nbsp;</li></ul><p>The <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=6495+LC%2FTQ"><strong>6495 LC/TQ</strong></a> with an Agilent&nbsp;Jet Stream (AJS) electrospray ion source was operated in dynamic multiple reaction monitoring (dMRM) mode. This mode allows more MRM transitions if future development is required for additional compounds. The LC/TQ autotune was performed in both unit and wide modes. All data acquisition and processing were performed using Agilent&nbsp;MassHunter software (version&nbsp;12).</p><h4><strong>Conclusion&nbsp;</strong></h4><p>This application note describes a highly sensitive and reproducible method for the fast and reliable quantitation of the listed compounds in water by direct injection. The dMRM method was created and optimized using Agilent MassHunter software and allows the addition of more MRM transitions if future development is required for additional compounds. An Agilent 1290 Infinity II LC coupled to an Agilent 6495D LC/TQ was used for the analysis. The 12-minute LC gradient method using an Agilent Aq-C18 column offered good chromatographic separation and RT distribution of all targets. The LC/TQ data acquisition was in dMRM mode with fast polarity switching for the most efficient use of instrument cycle time. The method performance was verified based on requirements for calibration curve linearity, instrument LOD, recovery, and precision. The results demonstrate the ability of the quantitative analytical method for emerging contaminants in water by direct injection.</p><h3><strong>2. Shimadzu: Short-Chain Oligonucleotide Analysis Using Supercritical Fluid Chromatography</strong></h3><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/ap_aplnote100_en_23b7bffc7e.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Oligonucleotide therapeutics are drugs in which oligonucleotide is the active ingredient. Oligonucleotide therapeutics are manufactured through chemical synthesis, but impurities are produced during the process, so measurement and characterization of the impurities are required. Currently, reversed-phase ion-pair chromatography is often used for this analysis, but it is difficult to separate some impurities that are structurally similar to the target product. Therefore, a separation method with selectivity different from existing methods is required.&nbsp;</p><p>SFC is an analytical method that employs carbon dioxide as the mobile phase at temperatures and pressures above the respective critical points. Compared to liquids, carbon dioxide is reported to have high resolution and special selectivity as a component of SFC mobile phase because of its low viscosity and high diffusivity.&nbsp;</p><p>On the other hand, since the mobile phase containing carbon dioxide has low polarity, SFC was considered unsuitable for the analysis of highly polar compounds. However, in recent years, the composition of the modifier mixed with carbon dioxide and the choice of stationary phase have made it possible to analyze highly polar compounds,such as peptides and nucleotides2).&nbsp;</p><p>In this study, we evaluated the applicability of SFC to oligonucleotide analysis. Single-strand oligonucleotides of around 20-mer are often used as oligonucleotide therapeutics, but due to the extremely high polarity of the molecules and the difficulty of analysis, we first used 4-mer oligonucleotides for evaluation and investigated their basic retention behaviors.</p><h4><strong>Experimental</strong></h4><ul><li><strong>System:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=LCMS-9030"><strong>LCMS-9030</strong></a></li></ul><h4><strong>Conclusions&nbsp;</strong></h4><p>To evaluate the applicability of SFC for oligonucleotide analysis, we developed a separation method using 4-mer oligonucleotides with varying PS contents as model compounds and investigated their basic retention characteristics. First, we identified conditions that allowed oligonucleotides to elute, finding that a methanol-water mixture containing 50 mmol/L ammonium formate enabled elution and peak detection. Using these conditions, we analyzed T4 with varying PS contents on 12 columns, finding good separation with the Diol II column. To apply this method to more polar sequences, we re-evaluated modifier conditions and found that adding aminoethanol improved peak shape. Applying the optimized method to 14 oligonucleotide sequences showed that sequences with two or fewer G or C bases were suitable for this method. Additionally, higher-polarity sequences showed stronger retention, while oligonucleotides with varying PS contents showed weaker retention with higher polarity, indicating unique retention behavior for PS modifications. The high selectivity for sequences with different PS contents suggests that SFC can be applied to separating impurities with a PS linkage replaced by a PO linkage that can be included in PS-modified oligonucleotides in the synthesis process.&nbsp;</p><p>In the future study, the applicability of SFC to longer oligonucleotidesthan 4-mer will be evaluated.</p><h3><strong>3. Thermo Fisher Scientific / HPLC Symposium: Sensitive LC-MS method for the quantitative analysis of semaglutide and liraglutide in human plasma</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_sensitive_lc_ms_method_quantitative_analysis_semaglutide_po_en_592e7dcc4a.pdf"><strong>Full PDF for download</strong></a></li></ul><p>GLP-1 analogs have emerged as pivotal therapeutic agents in the management of type 2 diabetes and obesity. These analogs mimic the physiological actions of endogenous GLP-1, including enhancing glucose-dependent insulin secretion, suppressing glucagon release, delaying gastric emptying, and promoting satiety. Given their significant clinical utilities, accurate detection and quantification of GLP-1 analogs in human plasma are essential for pharmacokinetic studies, therapeutic monitoring, and ensuring patient safety. This study aims to develop a sensitive LC-MS method for the detection and quantitation of semaglutide and liraglutide in human plasma.</p><h4><strong>Materials and methods</strong></h4><h5><strong>Mass Spectrometry</strong></h5><p><span>&nbsp;</span>See Table 2 for detailed MS source and scan settings for the analysis GLP-1 analogs on <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/404"><strong>TSQ Altis Plus MS</strong></a>.&nbsp;</p><h5><strong>Data Analysis&nbsp;</strong></h5><p><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/74"><strong>Thermo Scientific Enterprise </strong></a>compliance ready Chromeleon CDS 7.3.2 software was used for all instrument control, data acquisition, processing, and reporting.</p><h4><strong>Conclusions&nbsp;</strong></h4><p>We demonstrated a comprehensive LCMS solution for highly sensitive quantitation of GLP-1 analogs in human plasma using the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/744"><strong>Vanquish UHPLC</strong></a> coupled to TSQ Altis Plus MS. The solution enables:&nbsp;</p><ul><li>Robust quantitation of semaglutide and liraglutide with a LOQ of 0.1 ng/mL&nbsp;</li><li>Excellent precision and accuracy across 3 orders of LDR for the analysis of GLP-1 analogs&nbsp;</li><li>Minimal carryover observied with post-injection signals remaining below 10% of the LOQ for semaglutide and below 14% for liraglutide following the highest concentration injection (100 ng/mL)</li></ul><h3><strong>4. Waters Corporation: Method Migration of a Reversed Phase TFAAcetonitrile Gradient Method From a Binary to a Quaternary System: Impact of Mixing</strong></h3><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/720008689_en_51b4c42ae8.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Solvent mixing is critical to obtaining optimal gradient LC separations. The most commonly available pump mixing designs for reversed phase gradient HPLC separations are high pressure (typically binary) and low pressure (quaternary) systems. High pressure systems use independent pumps to deliver different solvents. Mixing occurs at high pressures with solvent composition controlled by the flow rates of the pump heads. In low pressure systems, solvent composition is controlled by a gradient proportioning valve. Each solvent is delivered in packets (based on the gradient specified in the method) which are mixed as they go through the pump head. High pressure systems have reduced dwell volumes and generally have improved compositional accuracy (leading to better retention time precision) over low pressure systems. Both high- and low-pressure systems are subject to mobile phase composition fluctuations. Mixers of various volumes and designs are utilized to help minimize these composition fluctuations by reducing baseline noise and oscillations in mobile phase composition.&nbsp;</p><p>The use of certain mobile phase modifiers is known to impact chromatographic baselines. Trifluoroacetic acid (TFA) is a commonly used ion-pairing reagent used in combination with acetonitrile for many gradient reversed phase applications. TFA absorbs strongly at wavelengths below 250 nm. Additionally, TFA is slightly retained on reversed phase columns which results in fluctuations in TFA concentration as the acetonitrile gradient passes through the column. In combination, these factors result in significant baseline disturbances (ripples) when TFAacetonitrile gradients are used at low wavelengths. These baseline ripples may make peak integration difficult and can impact the sensitivity along with peak area precision and retention time precision of the method.&nbsp;</p><p>Most modern HPLC systems typically include a standard mixer in the pump design. Additionally, different mixers may be available to improve mixing performance for specific applications. The effect that mixers have on chromatographic performance was demonstrated during the migration of a reversed phase TFA-acetonitrile gradient method from a binary system to the quaternary Alliance iS HPLC System.</p><h4><strong>Experimental</strong></h4><ul><li><strong>LC systems: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/717"><strong>Alliance iS HPLC System</strong></a> Comparable Binary HPLC system (System X)&nbsp;</li><li><strong>Detection: </strong>UV Detector with 10 mm HPLC flow cell</li><li><strong>Column: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1823"><strong>XBridge™ C18 Column</strong></a>, 5 µm 4.6 x 250 mm (p/n: 186003117)</li></ul><h5><strong>Data Management&nbsp;</strong></h5><ul><li><strong>Chromatography data system: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/627"><strong>Empower™ 3.8.0.1</strong></a></li></ul><h4><strong>Conclusion&nbsp;</strong></h4><p>The differences in mixing performance between binary and quaternary HPLC Systems may impact method migration. Mixing performance is particularly important when migrating gradient TFA-acetonitrile methods at low wavelengths, as these methods are known to produce significant baseline noise (ripple). In this study, effect of mixers on chromatographic performance was demonstrated through the migration of the USP Tryptophan Organic Impurities method from a binary system to the quaternary Alliance iS HPLC System. The results show that the characteristic baseline ripple associated with TFA-Acetonitrile gradients at low wavelengths is impacted by the choice of mixer. The Alliance iS HPLC System with the 690 µL Ti diffusion bonded mixer displayed a significant reduction of baseline noise when compared against both the binary and quaternary systems equipped with their respective standard mixers. This resulted in easier peak integration, improved peak area and retention time reproducibility, and increased method sensitivity.</p>

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