<h3>Two Dimensions, One Solution: Advancing Oligonucleotide Analysis with 2D-LC</h3><h4>Why Consider 2D-LC for Oligonucleotide Analysis?</h4>In oligonucleotide analytics, achieving reliable and precise results is not optional—it is fundamental. These biomolecules are inherently complex, often containing truncated sequences, structural variants, and a wide range of impurities. Such complexity frequently exceeds the separation capabilities of conventional one-dimensional liquid chromatography (1D-LC). As a result, critical components may coelute or remain unresolved, limiting analytical confidence.Two-dimensional liquid chromatography (2D-LC) offers a powerful alternative by significantly enhancing separation performance. By combining two complementary separation mechanisms, 2D-LC improves resolution, sensitivity, and overall analytical reliability. This enables a much deeper understanding of oligonucleotide samples and their compositional intricacies.<blockquote><h4>Previous parts from the Oligos Made Easy series</h4><ul><li>Part 1: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/article/6808">Overview</a></li><li>Part 2: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/article/6846">AEX</a></li><li>Part 3: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/article/6880">IP-RP</a></li><li>Part 4: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/article/6912">HILIC</a></li><li>Part 5: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/article/6944">SEC</a></li><li>Part 6: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/article/6982">Column Navigator</a></li><li>Part 7: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/article/7012">Large Scale AEX</a></li><li>Part 8: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/article/7045">Sepapure oliGO</a></li></ul></blockquote><h4>Moving Beyond the Limits of 1D-LC</h4>For many years, 1D-LC has served as a standard tool for biopharmaceutical characterization. However, its inherent limitations become apparent when dealing with highly complex mixtures. The selectivity of a single chromatographic dimension is restricted, and the lack of orthogonality in retention mechanisms often results in insufficient separation. As sample complexity increases, the resolving power of 1D-LC becomes inadequate.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_1_2_D_LC_Principle_Graphic_by_KNAUER_43e04d32a2.jpg" alt="KNAUER: Figure 1 - 2D-LC Principle (Graphic by KNAUER)." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_1_2_D_LC_Principle_Graphic_by_KNAUER_43e04d32a2.jpg 245w," sizes="100vw" width="1291" height="391">Two-dimensional LC addresses these limitations by coupling two chromatographic separations, which can be either similar or fundamentally different. Each dimension targets different physicochemical properties of analytes, enabling the resolution of compounds that would otherwise overlap. In essence, 2D-LC introduces an additional level of separation capacity, allowing analysts to disentangle coeluting species that remain unresolved in a single dimension.An important practical consideration is that the overall analysis time is governed by the second dimension. Because fractions from the first dimension must be processed sequentially, the second dimension must operate rapidly and efficiently. High-speed separations in the second dimension are therefore essential to maintain system performance and data quality.<h4>Architecture of a 2D-LC System</h4>At its core, a 2D-LC system relies on precise coordination between its components. The setup typically consists of two chromatographic columns, two pumping systems, and a switching valve that enables fraction transfer between dimensions. The sample is introduced through an autosampler and first separated on the primary column. Fractions of interest are then transferred into a loop via a switching valve and subsequently introduced into the second column for further separation.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_2_2_D_LC_System_Configuration_Example_Graphic_by_KNAUER_f5747adf3a.jpg" alt="KNAUER: Figure 2 - 2D LC System Configuration Example (Graphic by KNAUER)." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_2_2_D_LC_System_Configuration_Example_Graphic_by_KNAUER_f5747adf3a.jpg 245w," sizes="100vw" width="859" height="517">The valve plays a crucial role in this process, acting as the interface between both chromatographic dimensions. Its function is to alternately collect fractions from the first dimension and inject them into the second dimension under high-pressure conditions. Various valve configurations can be employed, including ten-port or eight-port valves with dual-loop arrangements, or synchronized six-port valves. Regardless of configuration, the underlying principle remains the same: while one loop is being filled, the other is being flushed into the second dimension.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_3_Schematic_figure_of_ten_port_left_and_eight_port_valve_right_2_position_setup_for_2_D_LC_Graphic_by_KNAUER_9209661ea8.jpg" alt="KNAUER: Figure 3 - Schematic figure of ten-port (left) and eight-port valve (right) 2-position setup for 2D-LC (Graphic by KNAUER)." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_3_Schematic_figure_of_ten_port_left_and_eight_port_valve_right_2_position_setup_for_2_D_LC_Graphic_by_KNAUER_9209661ea8.jpg 245w," sizes="100vw" width="1287" height="406">This alternating process requires precise timing. The second-dimension separation must be completed before the next fraction is ready for transfer. Consequently, synchronization between flow rates, loop volumes, and switching intervals is essential for maintaining chromatographic integrity.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_4_Schematic_figure_KNAUER_eight_port_2_position_valve_for_2_D_LC_Graphic_by_KNAUER_f9a5f06f5f.jpg" alt="KNAUER: Figure 4 - Schematic figure KNAUER eight-port 2 position valve for 2D-LC (Graphic by KNAUER)." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_4_Schematic_figure_KNAUER_eight_port_2_position_valve_for_2_D_LC_Graphic_by_KNAUER_f9a5f06f5f.jpg 245w," sizes="100vw" width="756" height="391"><h4>Applying 2D-LC to Oligonucleotide Analysis</h4>In oligonucleotide workflows, 2D-LC combines two complementary separation strategies to address sample complexity. The first dimension is typically used for bulk separation based on properties such as size, charge, or polarity. Techniques such as size-exclusion chromatography (SEC), anion-exchange chromatography (AEX), ion-pair reversed-phase chromatography (IP-RP), hydrophilic interaction chromatography (HILIC), or mixed-mode chromatography (MMC) are commonly applied. This step reduces sample complexity and can also serve as an online desalting stage, removing salts and buffer components that could interfere with downstream detection, particularly when mass spectrometry is used.Following this initial separation, selected fractions are transferred to the second dimension. This transfer can be performed either selectively or comprehensively, depending on the analytical goal. The second dimension is typically optimized for high-resolution separation, allowing detailed characterization of closely related species such as truncated oligonucleotides, sequence variants, or chemically modified impurities. When coupled with mass spectrometry, this setup enables accurate identification and sequencing.A key advantage of 2D-LC lies in the flexibility of combining different stationary phases and mobile phase conditions.&nbsp;Numerous separation combinations are possible, including:<ul><li>SEC×AEX</li><li>HILIC×IP-RP</li><li>IEC×IP-RP</li><li>IP-RP×IP-RP</li><li>AEX×HIC</li><li>MMC×SEC,</li><li>and even chiral×RP systems.&nbsp;</li></ul>This adaptability allows tailored workflows for specific analytical challenges.<h4>Heart-Cutting vs. Comprehensive 2D-LC</h4>When implementing 2D-LC, two primary transfer strategies are commonly used: heart-cutting and comprehensive transfer. Each approach offers distinct advantages depending on the analytical objective.Heart-cutting 2D-LC focuses on selected regions of interest from the first dimension. Only specific peaks or time windows are transferred to the second dimension for further analysis. This approach allows extended separation time and higher resolution in the second dimension, making it particularly suitable for targeted investigations of known critical compounds. However, because only selected fractions are analyzed, unexpected impurities outside the selected region may remain undetected.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_5_2_D_LC_Single_Heart_Cut_Principle_Graphic_by_KNAUER_b1b400784b.jpg" alt="KNAUER: Figure 5 - 2D-LC Single Heart-Cut Principle (Graphic by KNAUER)." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_5_2_D_LC_Single_Heart_Cut_Principle_Graphic_by_KNAUER_b1b400784b.jpg 245w," sizes="100vw" width="1276" height="319">In contrast, comprehensive 2D-LC transfers the entire eluate from the first dimension into the second dimension in a continuous manner. This approach provides a complete and detailed representation of the sample, making it ideal for research and development, impurity profiling, and regulatory applications. While more demanding in terms of speed and system performance, comprehensive 2D-LC ensures full sample coverage.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_6_Full_Comprehensive_2_D_LC_Principle_Graphic_by_KNAUER_e13da63ef1.jpg" alt="KNAUER: Figure 6 - Full Comprehensive 2D-LC Principle (Graphic by KNAUER)." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_6_Full_Comprehensive_2_D_LC_Principle_Graphic_by_KNAUER_e13da63ef1.jpg 245w," sizes="100vw" width="1327" height="292">In practice, hybrid approaches may also be employed, combining targeted heart-cutting with broader comprehensive analysis to balance resolution and coverage.<h4>Practical Considerations for Method Development</h4>Successful implementation of 2D-LC for oligonucleotide analysis requires careful method design.&nbsp;<h5>1. Column selection&nbsp;</h5>It is a critical starting point, as the combination of stationary phases must provide sufficient orthogonality while maintaining compatibility between mobile phases. Typically, the first dimension is optimized for general separation of oligonucleotide size or class, while the second dimension focuses on sequence-specific resolution and compatibility with mass spectrometry.<h5>2. Mobile phase selection&nbsp;</h5>Differences in pH and organic modifiers can significantly influence separation behavior, but incompatibility between dimensions can compromise performance. Therefore, mobile phases must be designed as a coordinated system. The use of volatile buffers, such as ammonium acetate, is often preferred for MS-based detection.<h5>3. Instrumentation&nbsp;</h5>The switching valve must operate reliably under high pressure, with precise timing to ensure accurate fraction transfer. Loop volumes must be carefully selected to avoid overfilling, as excessive loading can lead to sample loss and distortion due to flow profile effects. As a general guideline, loops should not be filled beyond approximately 70% of their capacity.<h5>4. Method development&nbsp;</h5>Typically begins with optimization of the second dimension, as it dictates overall system timing. Fast separations using short columns and steep gradients are often required to maintain sufficient sampling frequency. Once the second dimension is established, the first dimension can be adjusted to achieve optimal resolution and compatibility.<h4>Final Remarks</h4>Two-dimensional liquid chromatography represents a powerful advancement in oligonucleotide analysis, enabling the resolution of complex mixtures that exceed the capabilities of conventional 1D approaches. By combining orthogonal separation mechanisms, 2D-LC reveals structural details and impurity profiles that would otherwise remain hidden.However, achieving optimal performance requires careful balancing of multiple factors, including column chemistry, mobile phase compatibility, system timing, and data acquisition. When properly implemented, 2D-LC provides a robust and highly informative platform for modern oligonucleotide characterization.<blockquote><ul><li>If you're planning to set up or optimize your oligonucleotide workflow, feel free to contact us at <a href="sales@knauer.net">sales@knauer.net</a>. 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 <a href="kindler@knauer.net">kindler@knauer.net</a></li></ul></blockquote>

Oligos Made Easy - Part 9: 2D-LC

News from LabRulezLCMS Library - Week 14, 2026

<h3>We're excited to announce that <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/999">Clarity Chromatography Software</a> version 10.1.2 is now available!</h3>This minor update expands Clarity’s portfolio with new control modules.<h4>Newly added Instrument control modules</h4><ul><li>Ecom auxiliary modules</li><li>HTA ICF control modules</li><li>Spark pump</li></ul><h4>Updated Instrument control modules</h4><ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/companies/168">Ecom</a> HPLC</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/companies/328">Knauer</a> HPLC</li><li>Spark ICF control modules</li><li>YoungIn Chromass HPLC</li></ul><blockquote>For a complete list of module changes, see the <a target="_blank" rel="noopener noreferrer" href="https://www.dataapex.com/version_history?product=1">Changelog</a>. For more info about exciting new features directly in Clarity, explore what was introduced in the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/article/5622">Clarity 10.1 release highlights</a>.Clarity can control 1000+ instruments; the <a target="_blank" rel="noopener noreferrer" href="https://www.dataapex.com/controls">list can be previewed here</a>.</blockquote><h4>Supported Instruments in Clarity (DataApex) — Quick Guide</h4>Clarity Chromatography Software supports a wide range of chromatographs and peripherals via digital control (DD) and analog acquisition (A/D) using the Colibrick module.<h5>Gas Chromatographs (GC)</h5>Digital integration over LAN/RS232 (many via ICF):<ul><li>Agilent: 5890, 6890, 7890, 8860, 8890, Intuvo 9000</li><li>Shimadzu: GC-2010/2014, Nexis GC-2030</li><li>Thermo Fisher: TRACE 1300/1310</li><li>PerkinElmer: Clarus 400–690</li><li>Also supported: Dani, Ellutia, Young In Chromass, Apix, Fuli, GOW-MAC (selected models)</li></ul><blockquote>Note: Most Agilent 6890–8890 and Intuvo use LAN – ICF.</blockquote><h5>HPLC Systems</h5>Full system/module control:<ul><li>Agilent 1100–1290 Infinity II (ICF)</li><li>Shimadzu LC-10/20 and Nexera LC-40</li><li>PerkinElmer Flexar / NexSAR</li><li>Knauer Azura / BlueShadow / Smartline</li><li>Hitachi Chromaster / Primaide</li><li>Young In Chromass ChroZen series</li><li>Sykam S500–S600, Watrex Streamline, Hanon LC7000–7300</li></ul><h5>HPLC Modules (Pumps, Detectors, Ovens, Fraction Collectors)</h5><ul><li>Pumps: Agilent 1100–1290 families, Knauer, Shimadzu, PerkinElmer, Sykam, SSI, Ecom</li><li>Detectors: VWD/DAD(PDA)/RID/FLD/ELSD across Agilent, Shimadzu, Knauer, PerkinElmer, Young In Chromass, Ecom, etc.</li><li>Column Ovens: Agilent G1316x / G7116x, Shimadzu CTO-20/30/40, Knauer CT2.1, PerkinElmer Flexar</li><li>Fraction Collectors: Knauer FC 6.1, Teledyne FOXY series, Ecom ECF 2096, Waters Fraction Collector II</li></ul><h5>Autosamplers</h5><ul><li>Agilent GC ALS (7673/7683/7693/8697), HPLC ALS modules (G1313A–G7167C)</li><li>Shimadzu AOC-20 / AOC-5000+, SIL-10 / SIL-20 / SIL-40 series</li><li>CTC Analytics PAL, HTA autosamplers, Spark Alias/UHPLC, Knauer AS 6.1L</li><li>PerkinElmer Flexar / NexSAR AS</li></ul><h5>Other Devices</h5><ul><li>GC×GC modulators (e.g., Charles University GC×GC Modulator)</li><li>Valves &amp; switch modules (Agilent, Knauer, Shimadzu, VICI-Valco)</li><li>Degassers, flow monitors, managers, interfaces</li><li>LC-MS / GC-MS: Advion Expression CMS, Microsaic 4500 MiD, Markes BenchTOF (and others)</li><li>Balances (EA extension): Mettler Toledo, Sartorius</li></ul><h5>Analog Acquisition (Colibrick)</h5>A/D support for legacy or analog-output devices:<ul><li>GC: Agilent 5880/4890A, Varian 3xxx/3800/3900, PerkinElmer Clarus series, Thermo TRACE, Shimadzu GC-14/9A, Scion 43x/45x/83xx/85xx, etc.</li><li>HPLC Detectors: Waters 2487/2489, ESA Corona, Hitachi L-series, JASCO FP/UV, Ecom LCD series, many others</li><li>Elemental &amp; Amino Acid Analyzers: Carlo Erba/Costech ECS, Biochrom 20/30+</li></ul>If you are interested in a DEMO VERSION, contact us!

DataApex Clarity Chromatography software 10.1.2 just released!

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Since 1991 DataApex develops and manufactures chromatography data stations. DataApex is constantly striving to provide users with efficient and up-to-date tools for chromatography data processing.

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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.

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N-linked glycosylation is a critically important and complex post-translational modification.
The structure of N-linked glycans can influence the function of glycosylated biotherapeutics, making the characterization of N-glycans an essential part of the therapeutic development process.
In this The Lab Central Premiere, learn about Agilent&#39;s complete offering of glycan sample preparation and analysis consumables to support a comprehensive end-to-end N-glycan workflow solution.
<blockquote>
Presenter: Dr. Ashish Pargaonkar (Lead for Biopharma Workflow Commercialization, Agilent Technologies)
</blockquote>
<blockquote>
Presenter: Ms. Paramjeet Khandpur (Biopharma Workflow Commercialization Lead, Agilent Techologies)
</blockquote>

Agilent Technologies: The Glycan Connection

The Glycan Connection

Learn about Agilent's complete offering of glycan sample preparation and analysis consumables to support a comprehensive end-to-end N-glycan workflow solution.

This online event has been designed for our customers and brings together opinion leaders, top scientists and industry innovators to share their research in the biopharma field.
The technologies presented will be CE and LC-MS.
Agenda:
<h5>10:00–10:10 Welcome and introduction</h5>
<ul>
<li>Steve Lock, SCIEX</li>
</ul>
<h5>10:10–10:35 Use of LC-MS in process analytical testing as part of development of a continuous manufacturing workflow</h5>
<ul>
<li>Daniel Myatt / Ronan Gough, CPI</li>
</ul>
<h5>10:35–11:00 Tips and tricks for biotherapeutics characterization by peptide mapping</h5>
<ul>
<li>Giulia Calloni, SCIEX</li>
</ul>
<h5>11:00–11:25 Developing methods for SEC-MS and IEX-MS for the analysis of biotherapeutics</h5>
<ul>
<li>Sarah Nguyen, Strasbourg University</li>
</ul>
<h5>11:25–11:35 Break</h5>
<h5>11:35–12:00 The power of CZE analysis for characterization of NANOBODY® molecules</h5>
<ul>
<li>Dries Dejaegere, Sanofi</li>
</ul>
<h5>12:00–12:25 Multi-capillary gel electrophoresis to increase throughput for the purity analysis of biotherapeutic proteins</h5>
<ul>
<li>Theresa Merlino, The Janssen Pharmaceutical Companies of Johnson &amp; Johnson</li>
</ul>
<h5>12:25–12:50 A disruptive approach to the characterization of mAb charge variants during biopharmaceutical process development</h5>
<ul>
<li>Greg Adams, FUJIFILM</li>
</ul>
<h5>12:50–13:00 Wrap-up and Q&amp;A</h5>
<blockquote>
Presenter: Steve Lock, PhD (Snr. Marketing and Market Development Manager, Biopharma, EMEAI, SCIEX)
</blockquote>
<blockquote>
Steve obtained his PhD in Physical Organic Chemistry from the University College of Swansea in 1993 and is a chartered chemist and a member of the Royal Society of Chemistry. He has worked for SCIEX in various technical roles for over 20 years and has presented at over 80 international meetings around the world. He currently works closely with customers in the biopharmaceutical industry. Steve is also currently an Adjunct Professor at the University of Pannonia.
</blockquote>
<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/SCIEX_agenda_day_1_6818ee0331/SCIEX-agenda-day-1.jpg" alt="SCIEX - agenda day 1.jpg"/>

SCIEX: SCIEX Biopharma meeting - 3rd edition! Day 1 - Gene therapy, vaccines and oligonucleotides

SCIEX Biopharma meeting (Day 1) - Gene therapy, vaccines and oligonucleotides

Gene therapy, vaccines and oligonucleotides. The technologies presented will be CE and LC-MS.

Day 1 - Gene therapy, vaccines and oligonucleotides

This online event has been designed for our customers and brings together opinion leaders, top scientists and industry innovators to share their research in the biopharma field.
The technologies presented will be CE and LC-MS.
Agenda:
<h5>10:00–10:10 Welcome and introduction</h5>
<ul>
<li>Steve Lock, SCIEX</li>
</ul>
<h5>10:10–10:35 Particle seizing by CE</h5>
<ul>
<li>Hervé Cottet, Montpellier University</li>
</ul>
<h5>10:35–11:00 Application of CE for purity analysis in cell and gene therapy</h5>
<ul>
<li>Ryan Hylands / Luiza Chrojan, Pharmaron</li>
</ul>
<h5>11:00–11:25 Tips and tricks for success in CE analysis</h5>
<ul>
<li>Whitney Smith, SCIEX</li>
</ul>
<h5>11:25–11:35 Break</h5>
<h5>11:35–12:00 RNA payload analysis: development and analytical challenges</h5>
<ul>
<li>Jérémie Parot, SINTEF Industry</li>
</ul>
<h5>12:00–12:25 The use of CE for AAV analytics</h5>
<ul>
<li>Padraig Flaherty, MeiraGTx</li>
</ul>
<h5>12:25–12:50 Oligo analysis by LC-MS</h5>
<ul>
<li>Alexandra Bushby, Labcorp</li>
</ul>
<h5>12:50–13:00 Wrap-up and Q&amp;A</h5>
<blockquote>
Presenter: Steve Lock, PhD (Snr. Marketing and Market Development Manager, Biopharma, EMEAI, SCIEX)
</blockquote>
<blockquote>
Steve obtained his PhD in Physical Organic Chemistry from the University College of Swansea in 1993 and is a chartered chemist and a member of the Royal Society of Chemistry. He has worked for SCIEX in various technical roles for over 20 years and has presented at over 80 international meetings around the world. He currently works closely with customers in the biopharmaceutical industry. Steve is also currently an Adjunct Professor at the University of Pannonia.
</blockquote>
<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/SCIEX_agenda_day_2_b442145213/SCIEX-agenda-day-2.jpg" alt="SCIEX - agenda day 2.jpg"/>

SCIEX: SCIEX Biopharma meeting - 3rd edition! Day 2 - Therapeutic proteins and peptides

SCIEX Biopharma meeting (Day 2) - Therapeutic proteins and peptides

Therapeutic proteins and peptides. The technologies presented will be CE and LC-MS.

Day 2 - Therapeutic proteins and peptides

Oligonucleotide analysis is a critical aspect of molecular biology and therapeutic development, demanding robust and versatile chromatographic techniques for comprehensive characterization.
Several modes of chromatography can be employed to characterize these biomolecules, but when and how to best use each mode can be challenging.
This webinar aims to explore how various chromatographic techniques can be synergistically employed to achieve comprehensive oligonucleotide characterization. Aspects of ion-pair reversed phase chromatography (IP-RPC), hydrophilic interaction liquid chromatography (HILIC), ion exchange chromatography (IEC), and size exclusion chromatography (SEC) will be discussed and how best to employ each mode in oligonucleotide characterization. Several application examples will be utilized to showcase how each of these modes of chromatography can be best utilized to fully resolve and understand the composition of samples containing oligonucleotides.
At the conclusion of the webinar, the audience member will:
<ul>
<li>Understand the strengths and weaknesses of each mode of chromatography in characterizing oligonucleotides.
</li>
<li>Be familiar with key method parameters to adjust when fine-tuning oligonucleotide chromatographic assays
</li>
<li>Gain an appreciation for when to use each mode of chromatography when analyzing oligonucleotides
</li>
</ul>
<blockquote>
Presenter: Cory E. Muraco (Biomolecule Workflows Manager, Merck KGaA, Darmstadt, Germany)
</blockquote>
<blockquote>
Cory joined the company in 2013, starting in the Chemical Standards division, developing and validating analytical methods for testing the purity of certified reference materials using GC and LC methods. In 2022, Cory assumed his present role, where he is tasked with supporting the design, development, and execution of the marketing and R&amp;D strategies around bioanalytical strategy.
</blockquote>
<blockquote>
Presenter: Andrea Krumm (Product Manager, Analytical Columns, Tosoh Bioscience GmbH)
</blockquote>
<blockquote>
Andrea Krumm studied biotechnology before gaining a PhD in Cell and Cancer Biology. After four years as an applications specialist for optical analytical devices used in academic and pharmaceutical research, she joined Tosoh Bioscience GmbH in 2020 as a product manager for analytical columns. Andrea is responsible for gathering customer requirements, application areas and addressing these with new and existing products of the analytical columns line.
</blockquote>

Merck: Exploring Oligonucleotide Diversity: Insights From Multiple Modes of Chromatography

Exploring Oligonucleotide Diversity: Insights From Multiple Modes of Chromatography

How various chromatographic techniques can be synergistically employed to achieve comprehensive oligonucleotide characterization.

Oligonucleotide Diversity: Multiple Modes of Chromatography

In this webinar, our expert speakers will focus on the Karl Fischer titration process, quality assurance, and relevant regulations such as pharmacopeias to improve application accuracy.
It will be of particular interest to people developing Karl Fischer applications or qualifying Karl Fischer titrations, especially lab technicians, lab managers, and quality control or assurance managers.
By attending this webinar, you will gain insight into the following:
<ul>
<li>Overview of the Karl Fischer titration process
</li>
<li>Tips to optimize your Karl Fischer application performance
</li>
<li>How to fulfill Quality Assurance Aspects and get an insight into regulations such as United States Pharmacopeia (USP) and European Pharmacopoeia (Ph. Eur.)
</li>
<li>Understanding of the Official Medicines Control Laboratory (OMCL) guidelines concerning the Karl Fischer oven system
</li>
</ul>
<blockquote>
Presenter: Bettina Straub-Jubb (Global Product Manager, Karl Fischer Titration at the life science business of Merck KGaA, Darmstadt, Germany)
</blockquote>
<blockquote>
Bettina is the Global Franchise Product Manager and lead expert for titration at Merck Life Science KGaA based in Darmstadt, Germany. She has more than 20 years’ experience in the instrumental analysis business, especially with Karl Fischer titration, general titration, reference materials, and diagnostics. Her responsibilities include improving and simplifying titration processes, driving new technologies, and advancing applications and reagents. She always works closely with the QC and application laboratories for titration as well as with the R&amp;D department to develop titration continuously. Bettina is a graduate engineer and holds a degree in Chemical Technology from the University of Applied Sciences, Darmstadt, Germany, and a degree as a Medical Technical Laboratory Assistant from the Medical Technical Academy Esslingen, Germany.
</blockquote>
<blockquote>
Presenter: Thomas Hitz (Application Chemist, Mettler Toledo)
</blockquote>
<blockquote>
Thomas Hitz studied biochemistry at the University of Zurich and completed his PhD in 2003 at the Swiss Federal Institute of Technology Zurich in the Department of Material Sciences and Post Doctorate in 2006. He provides market support at Mettler Toledo and has more than 15 years&#39; experience in application development, sales, and customer training, and as a technical advisor for product development. He provides support for complex automation solutions and is an expert in automated Karl Fischer titration.
</blockquote>

Separation Science: How to Fulfil Regulatory Compliance with Your Karl Fischer Titration Process

How to Fulfil Regulatory Compliance with Your Karl Fischer Titration Process

Karl Fischer titration process, quality assurance, and relevant regulations such as pharmacopeias to improve application accuracy.

Regulatory Compliance & Your Karl Fischer Titration Process

During drug metabolite identification (Met ID) studies collision-induced dissociation (CID) is the most commonly used fragmentation technique for LC-MS experiments.
Many conjugation metabolites are found to have multiple bonding potentials to parent drugs. It is a great challenge to locate the bonding sites by CID due to the information lost by highly selective cleavage on these bonds. 
The novel electron-activated dissociation (EAD) is a recently developed technology on the SCIEX ZenoTOF 7600 system. From a single LC-MS/MS experiment CID and EAD data can be collected, providing information rich fragmentation spectra. The additional information can help in confident localization and identification of metabolites compared to CID alone. These fragments can be crucial to locate the metabolic modification sites, such as conjugations. 
In this study, the application of EAD in drug metabolite profiling is explored. The efficiency in the structural elucidation of conjugation metabolites is compared with that of CID.
Webinar Learning Objectives:
<ul>
<li>Learn how EAD can be advantageous in confident identification and localization of drug metabolites
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<li>Discover the sensitivity of the Zeno trap technology in the identification of the low abundant metabolites
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<li>Explore the capabilities of the Molecule Profiler software in processing EAD and CID data for drug metabolite identification
</li>
</ul>
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Presenter: Ming Yao (Principal Scientist, CPPDB, Bristol-Myers Squibb)
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Ming Yao currently works at Bristol-Myers Squibb (BMS). He received his B.Sc. from Henan University, and his M.Sc. from the Chinese Academy of Preventive Medicine China. After graduation, he worked at the Institute of Occupational Medicine as an associate professor.
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As a visiting Associate Professor, he conducted research for bioactivation of chemicals at Merck as well as the University of Western Ontario. After then, he joined BMS and he was responsible for designing and conducting in vitro and in vivo biotransformation studies for multiple programs to support both development and discovery projects. Most recently, he has also been involved in the development of protein drugs, such as the antibody drug conjugates (ADCs) and biotransformation of biologics.
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Ming has over 28 years of experiences in the pharmaceutical industry and has over 50 publications and 5 book chapters covering new technologies of mass spectrometry and its applications in drug metabolism.
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Presenter: Rahul Baghla (Manager Pharma Applications, SCIEX)
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Rahul Baghla, Manager Pharma Applications is a research scientist with 12+ years of experience in various applications of mass spectrometry delivering in Pharma/Biopharma/CRO industries. He is specialised in implementing new mass spectrometry technologies in bioanalytical/analytical method development/validation, drug discovery, ADME assays and clinical studies.
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He has supported various functions within pharma industry including nutrition, wellness, and oral care based clinical trial studies over the years. Rahul has master’s degree in pharmaceutical analysis, master’s in business administration and a professional diploma in clinical research.
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He is responsible for continuous engagement with pharmaceutical community to identify major analytical challenges, provide solutions and to find significant analytical opportunities for SCIEX.
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SCIEX: Discover how Bristol-Myers Squibb uses electron-activated dissociation (EAD) to confidently identify drug metabolites and localize site of metabolism.

Discover how Bristol-Myers Squibb uses electron-activated dissociation (EAD) to confidently identify drug metabolites and localize site of metabolism.

Learn how EAD can be advantageous in confident identification and localization of drug metabolites.

LC/MS EAD & drug metabolites & localization of metabolism

MALDI Imaging is a powerful tool that can not only provide stunning visuals of tissues but also serve as an important tool in answering biological questions.
Join Dr. Anand Mehta from the Medical University of South Carolina as he gives an overview of his work with imaging N-linked glycans from liver cancer tissues. He will share how this technology is being used to advance clinical research in disease prognosis, biomarker evaluation, and druggable targets.
This LabCast will also give you a front-row seat to see our experts demonstrate these techniques, so you can practice them in your own lab. There will also be an opportunity to ask questions at the end.
This LabCast demonstrates:
<ul>
<li>The principles of MALDI Imaging and how it is used to analyze N-linked glycans
</li>
<li>Sample preparation and data acquisition techniques
</li>
<li>Multiomic and multimodal data analysis strategies
</li>
</ul>
Who should attend:
<ul>
<li>Researchers interested in learning how to use MALDI Imaging to study N-linked glycans in tissues
</li>
<li>Anyone who is interested in learning more about MALDI Imaging and its applications in clinical research
</li>
<li>No prior experience with MALDI Imaging is required.
</li>
</ul>
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Presenter: Anand Mehta (DPhil, Professor, Endowed SmartState Chair in Proteomic Biomarkers, Medical University of South Carolina, Charleston, South Carolina, USA)
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Dr. Anand Mehta is a Professor at the Medical University of South Carolina, where his research work focuses on the glycobiology of liver cancer and the identification and development of glycan-based biomarkers for the early detection of cancer. Together with Dr. Rick Drake and Dr. Peggi Angel at MUSC, Dr. Mehta has pioneered the utilization of MALDI Imaging for the analysis of N-linked glycans utilizing multiple N-glycan-targeting enzymes including PNGase F. To date, Dr. Mehta has authored over 100 research articles on topics including the glycobiology of liver cancer and the identification of cancer biomarkers. Dr. Mehta has also been awarded multiple U.S. patents on liver pathology and glycan-based biomarkers of liver cancer. Dr. Mehta’s current work focuses on the development of high-throughput MALDI Imaging-based platforms for the identification and validation of cancer biomarkers in both tissue and serum samples.
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Presenter: Andrew DelaCourt (Ph.D., Field Application Scientist in Imaging, Bruker, Charleston, South Carolina, USA)
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Dr. Andrew DelaCourt is a Field Application Scientist at Bruker focusing on MALDI Imaging applications. Dr. DelaCourt recently completed his Ph.D. work in the laboratory of Dr. Anand Mehta at the Medical University of South Carolina, where he utilized MALDI Imaging for the identification of N-linked glycan biomarkers of genetic subtypes of hepatocellular carcinoma. Dr. DelaCourt brings his expertise in cancer biology and glycobiology to Bruker in order to further contribute to the application of MALDI Imaging in those areas.
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Presenter: Daniel Castro (Ph.D., Application Scientist in Imaging, Bruker, Billerica, Massachusetts, USA)
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Dr. Daniel Castro is an Application Scientist at Bruker focusing on MALDI Imaging. Dr. Castro completed his Ph.D. in the lab of Jonathan Sweedler at the University of Illinois-Urbana-Champaign, where he focused on high-throughput single-cell and single-organelle measurements as well as accelerating tissue MALDI-FTICR imaging. Dr. Castro, is an expert in high-throughput MALDI measurements.
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Bruker: LabCast: Biomarkers for liver cancer using N-Glycan MALDI Imaging

Biomarkers for liver cancer using N-Glycan MALDI Imaging

The principles of MALDI Imaging and how it is used to analyze N-linked glycans. Sample preparation and data acquisition techniques. Multiomic and multimodal

From applied research to corrosion monitoring to quality control of boiler feed water in the power industry – voltammetry is an established method for highly selective and sensitive analysis of heavy metals. Now, a new generation of mercury-free sensors developed by Metrohm makes this method «green» and hence compatible with increasingly tougher environmental regulations in many countries.
Join Jakub Tymoczko, Application Specialist at Metrohm International Headquarters, in his webinar on November 2 and learn how how you can perform «green voltammetry» in the laboratory as well as in the field for fast determination and speciation of heavy metals ranging from arsenic to zinc.
Key learnings of this webinar:
<ul>
<li>The principle behind the new generation of mercury-free sensors from Metrohm</li>
<li>The inherent benefits of voltammetry for heavy metal analysis such as lower costs, easier to use, faster results than by competing methods, e.g., ICP-MS</li>
<li>Typical application examples from different fields (research, process control, monitoring of drinking water)</li>
<li>Automation possibilities for high throughput analysis of large sample series</li>
</ul>
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Presenter: Jakub Tymoczko (Application Specialist Voltammetry, Metrohm International Headquarters)
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Dr. Jakub Tymoczko has worked for the Competence Center Voltammetry at Metrohm International Headquarters in Switzerland since 2015. In this time, he has been in charge of developing Metrohm&#39;s new generation of mercury-free sensors and methods for heavy metal detection. Jakub has a Master in biochemistry and received his PhD in electrochemistry on the engineering of model electrocatalysts for energy conversion and storage from the Ruhr-University in Bochum.
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Metrohm: Green voltammetry – a new generation of sensors for mercury-free analysis of heavy metals

Green voltammetry – a new generation of sensors for mercury-free analysis of heavy metals

How you can perform «green voltammetry» in the laboratory as well as in the field for fast determination and speciation of heavy metals.

Green voltammetry – mercury-free analysis of heavy metals

In this webinar, we will present end-to-end solutions for biopharma lab workflows with a focus on multiple mass spectrometry- based analysis techniques, ranging from SQ, TQ, TOF and Q-TOF.
A broad portfolio in sample prep automation for protein therapeutics allows for highly reproducible results in the starting point of the analysis. A broad range of consumables and columns will be presented, leading into the subsequent separation via HPLC and detection by mass spectrometry.
The 6545XT AdvanceBio was particularly developed for intact protein analysis, and the ultra-high vacuum and large molecule autotune led to excellent sensitivity in denatured and native applications, allowing quantitation on the intact protein level as well as ADR calculations. Peptide mapping and glycan analysis results, along with the confirmation of oligonucleotide sequences using Q-TOF-based workflows, will be presented.
We will also highlight the ultrahigh performance of the novel 6495 triple quadrupole LC/MS system (G6495D) with MassHunter Workstation 12.1 for comprehensive multiplexed protein quantification. Artificial intelligence (AI)-based SWARM autotune and enhanced system features provide seamless protein quantification method optimization, ultimate analytical sensitivity, and high analytical precision as well as user-friendly instrument care and maintenance.
Lastly, we will show results from oligonucleotides and proteins analyzed on LC/MS single quadrupole and Time of Flight (TOF) instruments, the latest scientific applications, and recent method improvements.
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Presenter: Ian Edwards, Ph.D. (LC/TQ Portfolio Product Manager, Agilent Technologies Inc.)
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Ian holds a PhD from the Centre for Proteome Research at the University of Liverpool. He has spent time as a pre- and post-sales MS applications specialist focused on MS imaging and proteomics, as a field marketing manager for -omics, and as a global pharmaceutical marketing manager. Ian joined Agilent recently as LC/TQ Product Manager.
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Presenter: Russell Burge, Ph.D. (LC/SQ and LC/TOF Portfolio Product Manager, Agilent Technologies Inc.)
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Russell earned a PhD in biochemistry and molecular biology from Scripps Research. He has extensive experience in the R&amp;D of biomolecules and biopharmaceuticals, and the implementation of automated workflows. Russell has acted as a Product Manager for various products in Life Science and Industrial Markets, and he currently manages the LC/MS SQ and LC/MS TOF product portfolios at Agilent.
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Presenter: Christian Klein, Ph.D. (LC/Q-TOF Portfolio Product Manager, Agilent Technologies Inc.)
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With an education in Chemistry, Chris Klein received his Ph.D. in 2005 at the Max-Planck-Institute for Biochemistry in Martinsried, Germany. He joined Agilent Technologies in 2010 as the lead chemist in the LC/Q-TOF R&amp;D team. In 2018 Chris jointed the marketing team as Portfolio Product Manager, responsible for LC/Q-TOF and Ion Mobility.
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Presenter: Róisín Murtagh (Editor, Wiley)
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Róisín Murtagh has been an editor for Wiley Analytical Science since 2021. She holds a science degree from Maynooth University, Ireland, and an MSc in Medicinal Chemistry from the Open University, UK. With a background encompassing over 10 years of experience in analytical science, she has actively contributed to both industry and academic research.
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Wiley: Intelligent LC/MS tools for quantitation and characterization of biopharmaceuticals

Intelligent LC/MS tools for quantitation and characterization of biopharmaceuticals

End-to-end solutions for biopharma lab workflows with a focus on multiple mass spectrometry- based analysis techniques, ranging from SQ, TQ, TOF and Q-TOF.**

LC/MS quantitation & characterization of biopharmaceuticals

The recent success of mRNA vaccines demonstrates the capabilities of nucleic acid-based drugs. These new modalities hold the promise of delivering not only potent vaccines, but also enzyme replacement therapies. However, there is a need for more analytical characterization tools to provide more informationon stability, heterogeneity, drug design and structure-function relationships. Anion-exchange (AEX) and size exclusion (SEC) chromatography techniques are well suited to providing high resolution separations of nucleic acids and their carriers.
AEX and SEC offer comprehensive quality and impurity profiling, but have suffered with previous methods from unsatisfactory recovery and resolution, poor MS compatibility. In addition, column hardware and packing materials have exhibited problematic secondary interactions. We have explored alternative mobile phase conditions in combination with various packing materials and sorbents in parallel with using novel low adsorption hardware allowing for more robust platform method for multiple drug substances and sample types, including AAVs and mRNA.
Learning Objectives Include:
<ul>
<li>Comprehensive overview of both theoretical and practical aspects of modern AEX and SEC separations.
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<li>New solutions and method parameter consideration for design of robust methods for nucleic acid separations.
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<li>Chromatography examples for small to very large nucleic acids
</li>
</ul>
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Presenter: Szabolcs Fekete PhD (R&amp;D Investigator, Waters Corporation)
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Dr. Szabolcs Fekete worked as an analytical R&amp;D scientist in the pharmaceutical industry for 10 years then moved to the University of Geneva and worked as a scientific collaborator for a decade. In April 2021, he joined Waters Corporation and now works as one of the Chemistry department’s lead application and development scientists. . He has contributed to 200 pieces of published work.
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Waters Corporation: New Insights on Anion Exchange and Size Exclusion of Nucleic Acids

New Insights on Anion Exchange and Size Exclusion of Nucleic Acids

We have explored alternative mobile phase conditions in combination with various packing materials and sorbents.