<h3>A Dedicated Solution for Oligonucleotide Analysis</h3>Oligonucleotides have become key players in modern pharmaceutical and biological research, particularly in the development of highly targeted and personalized therapies. As increasing numbers of oligonucleotide-based drugs enter clinical trials or receive regulatory approval, analytical laboratories face growing demands for both throughput and data quality. These molecules present unique chromatographic challenges due to their long, highly polar structures and subtle variations in nucleobase sequence or functionalization.<h3>Sepapure oliGO – Designed for Oligonucleotide Separations</h3>Selecting the optimal chromatographic approach for oligonucleotide analysis can be complex. Many laboratories rely on ion-pair reversed-phase (IP-RP) chromatography because of its speed, robustness, and compatibility with mass spectrometry. However, commonly used ion-pair reagents can degrade conventional silica-based stationary phases.<a target="_blank" rel="noopener noreferrer" href="https://www.knauer.net/columns-oligonucleotide-columns">Sepapure oliGO</a> addresses this limitation through a hybrid silica matrix combined with specialized surface end-capping, ensuring resistance to aggressive IP-RP reagents. Its functionalized surface enables both strong reversed-phase interactions and controlled ionic contributions, effectively creating a mixed-mode separation mechanism. This design results in improved durability compared with standard mixed-mode columns, making Sepapure oliGO a robust choice for routine oligonucleotide workflows.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_1_Sepapure_oli_GO_columns_with_laser_etched_column_information_23f545be81.jpg" alt="KNAUER: Figure 1 - Sepapure oliGO columns with laser-etched column information." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_1_Sepapure_oli_GO_columns_with_laser_etched_column_information_23f545be81.jpg 223w," sizes="100vw" width="1186" height="828"><h3>Method Transfer and Comparative Performance</h3>In collaboration with BianoGMP GmbH, KNAUER’s application team evaluated Sepapure oliGO against a widely used reference column. The study compared oligonucleotides of varying lengths and purities, including a fully thiolated 43-mer DNA. Particular attention was paid to potential interactions between thiolated oligonucleotides and stainless-steel hardware, which have been reported to cause peak tailing.The results demonstrated that direct method transfer between the reference column and Sepapure oliGO was generally feasible without significant adjustment. Chromatographic resolution remained comparable, while retention times were slightly shorter on Sepapure oliGO. Furthermore, the anticipated benefits of expensive bioinert hardware were not confirmed under the tested conditions, suggesting that such effects may primarily influence very fast (U)HPLC separations.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_2_Overlayed_chromatogram_of_crude_product_of_a_43mer_thiolate_oligonucleotide_7ae9613efd.jpg" alt="KNAUER: Figure 2: Overlayed chromatogram of crude product of a 43mer thiolate oligonucleotide. Light blue - reference column and dark blue – Sepapure oliGO." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_2_Overlayed_chromatogram_of_crude_product_of_a_43mer_thiolate_oligonucleotide_7ae9613efd.jpg 245w," sizes="100vw" width="1223" height="453"><h3>Low Backpressure for Reliable Operation</h3>Sepapure oliGO can be applied effectively in both HPLC and (U)HPLC workflows. Its low operating backpressure minimizes risks associated with clogging, excessive system wear, and performance deterioration over time. Typical IP-RP separations using 3 µm particles generated pressures around 200 bar, enabling laboratories without ultra-high-pressure systems to perform high-quality oligonucleotide analysis.<h3>Best Practices for Column Performance</h3>To maintain optimal performance, users should observe recommended operating conditions, including a broad pH range of 1.5–10 and temperatures up to 60 °C. Employing a guard column can prevent coarse synthesis by-products from reaching the analytical column. If pressure increases are observed, backflushing at low flow rates can restore performance.When the system is idle, thorough flushing to remove salts is essential. For storage, the column should be kept in a 1:1 mixture of methanol–water or acetonitrile–water. Gradient transitions are recommended when switching between standard RP eluents and IP-RP reagents. Regeneration procedures developed for RP columns are also suitable for Sepapure oliGO.<h3>Beyond Analytical Separations</h3>For large-scale purification, ion-exchange chromatography (AEX) may offer advantages. Scalable preparative systems are available to support purification workflows, ranging from compact laboratory setups to pilot-scale platforms with flow rates up to 800 mL/min. Bioinert configurations can be integrated where required, allowing laboratories to tailor purification solutions to their specific oligonucleotide applications.<blockquote><ul><li>If you're planning to set up or optimize your oligonucleotide workflow, feel free to contact us at sales@knauer.net. Stay tuned for more exciting insights into the Oligonucleotide world in our “Oligos Made Easy” series.</li><li>For more in-depth discussion or questions, reach out to the author at folmert@knauer.net&nbsp;</li></ul></blockquote>

Oligos Made Easy - Part 8: Sepapure oliGO

Visit the LabRulez booth at <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/events/871">analytica 2026</a> in Munich, 24. - 27. March 2026! You can find us in Hall A1, Booth A1.506. At the booth, you will be able to explore live demonstrations of how the LabRulez portals work and see selected analytical instruments from our pre-owned marketplace section.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Lab_Rulez_at_Analytica_2026_Floor_plan_55e1057e03.jpg" alt="LabRulez at Analytica 2026: Hall A1 Floor Plan and Booth Location" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Lab_Rulez_at_Analytica_2026_Floor_plan_55e1057e03.jpg 245w," sizes="100vw" width="1516" height="881"><h3>Would you like to learn how the LabRulez portals work?</h3>➡︎ <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/">LabRulezLCMS</a> ➡︎ <a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/">LabRulezGCMS</a> ➡︎ <a target="_blank" rel="noopener noreferrer" href="https://icpms.labrulez.com/">LabRulezICPMS</a><ul><li>A unique online solution for your marketing</li><li>A targeted audience from the analytical chemistry community</li><li>Modern technologies and a unique concept</li><li>Design optimized for most smart devices – PC, mobile, and tablet</li><li>Advanced statistical tools fully available to users</li></ul><h3>Services Offered</h3><h4>News + Company Updates</h4><ul><li>Publish news related to your instrumentation, applications, services, events, job openings, and more</li><li>News is shared both on our portals and across our social media channels</li></ul><h4>Library – All Documents in One Place</h4><ul><li>Applications, posters, presentations, technical articles, brochures, specifications</li><li>Fast and advanced search capabilities</li><li>Over 32,000 documents currently available in our database</li></ul><h4>Webinars</h4><ul><li>Upcoming and on-demand webinars from the field of analytical chemistry</li><li>More than 5,500 webinars covering chromatography, mass spectrometry, and spectroscopy</li></ul><h4>Careers</h4><ul><li>Targeted job listings in analytical chemistry</li><li>Clear filtering by specialization or location</li><li>Increased visibility for job postings</li></ul><h4>Companies</h4><ul><li>Company profile with key information and contacts</li><li>Integration of all your activities on our portal: News, Library documents, Webinars, Upcoming Events, Careers, and mentions by partners or customers</li><li>Connection to your Products listed on LabRulez portals</li></ul><h4>Products and Services</h4><ul><li>Comprehensive overview of instrumental techniques and services</li><li>Targeting the analytical chemistry community and customers in the growing online environment</li></ul><h4>Marketplace</h4><ul><li>Buying and selling pre-owned analytical equipment</li><li>We purchase analytical instrumentation of all brands and ages</li><li>Sell your equipment through LabRulez</li><li>Request and purchase complete systems, modules, or spare parts with warranty and service</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Lab_Rulez_at_analytica_2026_bazar_7edf4a0872.jpg" alt="LabRulez at analytica 2026" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Lab_Rulez_at_analytica_2026_bazar_7edf4a0872.jpg 88w," sizes="100vw" width="1600" height="2844"><h3>What Will Be on Display at Our Booth?</h3>Come and explore selected spare parts and modules from our marketplace section. These include both new, tested, and pre-owned items.<h4><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/bazaar">LabRulezLCMS Marketplace</a></h4>Spare parts for LC/MS:<ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/bazaar?category=378">Vacuum turbo pumps for LC/MS</a></li></ul>Spare parts for HPLC:<ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/bazaar?category=397">Degassers for HPLC/UHPLC</a></li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/bazaar?category=368&amp;search=valve">Automatic valves for HPLC</a></li><li>HPLC components for Agilent 1100, 1200, and 1260 Infinity II series</li></ul>Consumables:<ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/bazaar?category=377">HPLC lamps</a></li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/bazaar?category=48">HPLC columns</a></li></ul><h4><a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/bazaar">LabRulezGCMS Marketplace</a></h4>Spare parts for GC/MS:<ul><li><a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/bazaar?category=367&amp;search=turbo">Turbo pumps for GC/MS</a></li></ul>Spare parts for GC:<ul><li><a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/bazaar?category=366&amp;search=EPC">EPC modules</a></li></ul>Sample injection<ul><li><a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/bazaar/432">Autosampler 7693A for GC</a></li></ul>Consumables:<ul><li><a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/bazaar?category=7">GC columns </a>and selected accessories</li></ul>

LabRulez at analytica 2026

News from LabRulezLCMS Library - Week 13, 2026

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LabRulez

Based in Berlin, KNAUER is a medium-sized, owner-managed company that has been serving the sciences since 1962. We develop and manufacture scientific instruments of superior quality for liquid chromatography, including: Analytical HPLC/UHPLC, Preparative HPLC, Fast protein liquid chromatography (FPLC), Multi-column chromatography/Simulated moving bed (SMB) chromatography. High pressure dosing, in-line detection systems, and osmometry are additional business areas.

KNAUER

Determination of nitrosamine content (Bioanalytic)

Our offer includes comprehensive development and validation of methods for determination of nitrosamines in pharmaceutical products. We perform the work according to the criteria and needs of the client based on the EMA note.

Determination of nitrosamines is one of the pharmaceutical industry's top priorities. After the presence of N-nitrosodimethylamine (NDMA) was first reported in an active pharmaceutical ingredient (API), the U.S. Food and Drug Administration (FDA) and the European Medicines Agency (EMA) mandated the identification and control of all APIs and finished products in which there is a risk of nitrosamines.The content of nitrosamines in drug products can be controlled primarily by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) analysis. More than that, a reliable analysis result in our offer consists of a well-chosen sample preparation procedure, a selective and sensitive analytical method, and the experience of the analyst, who selects the best method parameters at the optimization stage.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Determination_of_nitrosamine_content_Bioanalytic_2_ce4f440837.jpg" alt="Bioanalytic: Determination of nitrosamine content." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Determination_of_nitrosamine_content_Bioanalytic_2_ce4f440837.jpg 234w," sizes="100vw" width="1536" height="1024">We have gained experience working with leading pharmaceutical companies. As part of our ongoing work, we have developed analytical methods for small-molecule nitrosamines of 70-150 g/mol. These compounds are mainly formed during manufacturing processes and depend on the presence and type of excipients (e.g. NDBA - N-nitroso-dibutylamine). We have also developed methods for determining molecules with higher nitrosamine drug substance related impurities (NDSRIs). In our work, we focus on achieving the highest possible sensitivity, selectivity and precision. To this end, we individually select the parameters of the methods depending on the composition of the drug and the expected nitrosamines. Due to the above criteria and the required detection limits, we perform nitrosamine impurity determinations using LC-MS/MS kits based on <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?manufacturerItems=52&amp;search=QTRAP+5500%2B">QTRAP 5500+</a> and <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?manufacturerItems=52&amp;search=QTRAP+6500%2B">QTRAP 6500+</a> mass spectrometers (<a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=55">SCIEX</a>).<blockquote>Our offerOur offer includes comprehensive development and validation of methods for determination of nitrosamines in pharmaceutical products. We perform the work according to the criteria and needs of the client based on the EMA note.At the stage of sample preparation, we focus on:<ul><li>obtaining the highest possible recovery of nitrosamines (matrix purification);</li><li>limiting additional synthesis of impurities (selection of appropriate equipment and reagents);</li><li>ensuring the stability of the sample prepared for LC-MS/MS analysis.</li></ul>During the optimization of the LC-MS/MS method, we solve such problems as:<ul><li>selection of an appropriate internal standard;</li><li>separation of the impurity from the API;</li><li>obtaining the expected sensitivity of the analytical method.</li></ul>Quantitative analysis of nitrosamines requires sensitivity, precision and accuracy, which we provide to our customers.</blockquote><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Determination_of_nitrosamine_content_Bioanalytic_1_d629e00e62.jpg" alt="Bioanalytic: Determination of nitrosamine content." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Determination_of_nitrosamine_content_Bioanalytic_1_d629e00e62.jpg 103w," sizes="100vw" width="1130" height="1710">

Identification of the proteins (Bioanalytic)

We perform proteomic analyses on a state-of-the-art high-resolution ZenoTOF 7600 (SCIEX), in which the Zeno ion trap and EAD (Electron-Activated Dissociation) technology together make a powerful combination.

Protein identification is based on experimental measurement of peptide molecular weights and matching the amino acid sequence of the protein under study with protein sequences deposited in libraries (databases). Two approaches are most commonly used in proteomic studies: top-down and bottom-up. The top-down method involves analyzing full protein sequences. Bottom-up analyses, on the other hand, use digestion of proteins with proteolytic enzymes. The peptides obtained by digestion are analyzed using a mass spectrometer. Each analysis leads to a comparison of experimental data with data obtained by theoretical digestion of proteins described in the database.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Identification_of_the_proteins_Bioanalytic_2_6231b73f0d.jpg" alt="Bioanalytic: Identification of the proteins." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Identification_of_the_proteins_Bioanalytic_2_6231b73f0d.jpg 245w," sizes="100vw" width="1536" height="766"><h4>LC-MS/MS</h4>Tandem mass spectrometers are commonly used to determine protein sequences. They allow ion selection of pseudomolecular peptides and then fragmentation and measurement of the masses of the resulting fragments. Fragmentation using collision induced dissociation(CID ) is an invaluable tool in targeted and untargeted MS/MS analysis of small molecules. It is important to note that in proteomic analysis, data obtained using the aforementioned fragmentation method may leave gaps. For example, spectra obtained by CID fragmentation do not allow unambiguous differentiation of some amino acids: leucine from isoleucine (identical molar masses and CID fragmentation) and lysine from glutamine (very similar masses). For this reason, we perform proteomic analyses on a state-of-the-art high-resolution <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?manufacturerItems=52&amp;search=+ZenoTOF+7600">ZenoTOF 7600</a> (SCIEX), in which the Zeno ion trap and EAD (Electron-Activated Dissociation) technology together make a powerful combination. It enables the creation of unique fragments to differentiate problematic amino acids and increase the sensitivity and specificity of the analysis.<blockquote>We perform analyses of:<ul><li>samples from solution;</li><li>samples from a gel;</li><li>samples from a complex matrix (by prior arrangement);</li><li>samples from lyophilizer;</li><li>samples previously prepared by the client.</li></ul>Qualitative analysis of proteins includes:<ul><li>To confirm the presence of a protein, the sample is subjected to digestion and the result obtained from experimental data is compared with theoretical data;</li><li>Comparison of experimental data on the masses of peptides included in the analyzed protein with the protein sequences described in the database;</li><li>Analysis and identification of the protein mixture;</li><li>Identification of post translational modifications (PTMs) (we use the tools: Skyline, Spectronaut).</li><li>Reporting of the result including, but not limited to, confirmation of the coverage of the protein sequence obtained in the test sample with the theoretical protein and the list of modified peptides, location and % of modifications.</li></ul>Comparative analyses include:<ul><li>Determining the composition of mixtures;</li><li>Comparison and profiling of sample series - protein identification and trend analysis.</li></ul>Our offer also includes:<ul><li>Support in the planning and execution of studies (the scope is determined individually);</li><li>Assistance in the development of extraction methods from complex matrices - biological fluids, plant tissue, animal tissues (the scope is determined individually);</li><li>Statistical analysis using tools: MarkerView (PCA), GraphPad Prism, Trends, Sciex OS;</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1294">Quantitative analysis of proteins</a>;</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1296">Analysis and characterization of monoclonal antibodies</a>.</li></ul></blockquote><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Identification_of_the_proteins_Bioanalytic_1_97cf8c94b3.jpg" alt="Bioanalytic: Identification of the proteins." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Identification_of_the_proteins_Bioanalytic_1_97cf8c94b3.jpg 117w," sizes="100vw" width="1500" height="2000">

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LabRulez at analytica 2026

Oligos Made Easy - Part 8: Sepapure oliGO

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