<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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Waters Corporation

<ul>
<li>Wait what?! Who knew!? There are so many things that a mass spec can do.
</li>
<li>I&#39;ve used it for years, yes this may be true.
</li>
<li>I&#39;ve processed some samples, more than a few.
</li>
<li>But between me and you, I had no idea all the things it could do!
</li>
<li>From sample prep to separation, you just won&#39;t believe…
</li>
<li>All of the tips and tricks we have up our sleeve.
</li>
<li>So, take a few minutes, about 15 should do.
</li>
<li>And learn all the things your mass spec can do.
</li>
</ul>
Thinking of developing a new method utilizing MS? Selecting the best analytical method begins with a decision on how to best separate the analytes of interest. This choice will affect sample preparation and sample introduction, so it is important to get it right early. In this webinar we will review analyte structural properties that make the compound more amenable to either liquid chromatography or to gas chromatography.
<blockquote>
Presenter: Karen Yannell (LC/MS Application Engineer, Agilent Technologies, Inc.)
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<blockquote>
Karen E. Yannell is a LC/MS Application Engineer at Agilent Technologies. She is an expert in small molecule high throughput analysis by triple quadrupole (TQ) and quadrupole time-of-flight (QTOF) mass spectrometry for applied markets. Her recent work has focused on developing routine Q-TOF workflows for both expert and non-expert users alike. Previously, Karen worked on forensic, metabolomics, and biomarker discovery applications with ambient ionization sources. She received her PhD in Analytical Chemistry from Purdue University where studied under the advisement of Prof. R. Graham Cooks. Outside of the laboratory, she enjoys cooking unique dishes and hiking along the beach with her dog.
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<blockquote>
Presenter: David Weil, PhD (Senior Applications Scientist, Agilent Technologies, Inc.)
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<blockquote>
David Weil is a senior applications scientist in Agilent located in Wood Dale with over 30 years of experience using mass spectrometry. Prior to joining Agilent in 2004, he worked for 14 years at 3M in the Corporate Research Analytical Laboratories (St.Paul, MN) and 4 years at Nicolet/Extrel doing ultra-high resolution mass spectrometry. David received his Ph.D. in Physical Chemistry from the University of Minnesota and Post Doc with Dr Charles Wilkins at University of California-Riverside developing laser desorption FTMS and ion-molecule reactions. David has three grown sons and live with his wife Naomi just blocks from Wrigley Field and Lake Michigan.
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<blockquote>
Presenter: Matt Curtis, PhD (GC/MS Applications Scientist, Agilent Technologies, Inc.)
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Matt joined Agilent in 2013 as a GC/MS applications scientist in the Santa Clara, CA Center of Excellence. His primary responsibility is focused on providing instrument demos for customer interested in each technology Agilent offers; the GC/MS single quadrupole, triple quadrupole and the quadrupole Time-of-Flight. The variety of different samples submitted to the demo lab provide a unique knowledge of optimizing parameters for different applications to provide the best quality data from the instrument.
</blockquote>

Agilent Technologies: Selecting your Separation: Are my Compounds Better for GC or LC

Selecting your Separation: Are my Compounds Better for GC or LC

We will review analyte structural properties that make the compound more amenable to either liquid chromatography or to gas chromatography.

Separation selection: Are my Compounds Better for GC or LC

TBD
<blockquote>
Presenter: Mark Sartain, Ph.D. (LC/MS Applications Scientist, Agilent Technologies, Inc.)
</blockquote>
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Mark has developed a diverse array of targeted and nontargeted mass-spectrometry based metabolomics and lipidomics applications since joining Agilent in 2012, with a focus on complete hardware/software workflows. Mark began utilizing mass spectrometry in his graduate work to characterize glycoproteins, and later established a novel LC/MS strategy to comprehensively profile and identify complex lipids of Mycobacterium tuberculosis in his postdoctoral fellowship at Colorado State University. He continued designing a wide range of MS-based techniques to measure metabolites and lipids for diverse projects as a Research Scientist at the Institute of Systems Biology (Seattle, WA) prior to joining Agilent.
</blockquote>

Agilent Technologies: LC/TQ Solutions to Give Your Lab Multi-omic Biological Insights

LC/TQ Solutions to Give Your Lab Multi-omic Biological Insights

LC/TQ to Give Your Lab Multi-omic Biological Insights

Many illicit and prescribed medications are reported to impair drivers control of their vehicles, increasing the potential for road traffic accidents.
Analytical testing to support DUID legislation requires the quantitation of large panels of drugs exhibiting a wide range of chemical properties. To achieve a simple workflow to detect all relevant molecules optimally and at their specified concentrations presents some analytical challenges.
This presentation will detail a fast, simple, and generic sample preparation method for whole blood and its application to DUID samples. The developed sample preparation method in combination with UPLC-MS/MS will be presented and the benefits of this protocol for laboratories will be discussed.
Learning objectives from the webinar – what you will learn: 
<ul>
<li>A background to driving under the influence – what it is, the importance, legislation, and analytical requirements.</li>
<li>Sample preparation protocol for whole blood samples and its application to DUID samples.</li>
<li>How a simple generic method could be beneficial to forensic toxicology laboratories involved in DUID analysis.</li>
</ul>
<blockquote>
Presenter: Emily Lee (Application Scientist, Forensics, Research &amp; Development, Waters Corporation, Waters Corporation)
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<blockquote>
Emily Lee is an Application Scientist at Waters Corporation. Emily is part of a global team dedicated to the development of applications and analytical solutions for Forensics and Toxicology. Emily has gained experience in the use of various Waters mass spectrometry technologies for the analysis of forensic and toxicologically relevant analytes. Prior to working at Waters, Emily has experience working in forensic toxicology laboratories, with high sample turnover and fast turnaround times. Emily was involved in many aspects of day-to-day laboratory operations including sample extraction, analysis, method development and validation, analytical troubleshooting, and reporting. Emily also gained a MSc in Forensic Science (Toxicology) at the University of Huddersfield.
</blockquote>

Waters Corporation: A Fast, Simple and Generic Sample Preparation Protocol for Whole Blood and its Application to DUID Samples

A Fast, Simple and Generic Sample Preparation Protocol for Whole Blood and its Application to DUID Samples

This presentation will detail a fast, simple, and generic sample preparation method for whole blood and its application to DUID samples.

Sample Preparation Protocol for Whole Blood and DUID Samples

Purification is critical to many scientific workflows. Compounds of interest are rarely found to be the right purity in their native state. Join Waters and Virscidian for this 6-part webinar series to learn the basics of purification and application of purification principles for more advanced workflows.
<h3>PART 4 | Using the Waters Fraction Manager Analytical (WFMA) to Achieve UPLC Scale Purification</h3>
Analytical scale purification provides for routine and robust isolation of your investigatory compounds. Challenges with UPLC purification include peak widths of 1-3 seconds and collecting the 6 to 50 µL peak volume. The Waters Fraction Manager Analytical (WFMA) addresses these challenges. Learn more about analytical scale purification with Waters. 
Learning Objectives: 
<ul>
<li>Understand analytical scale purification</li>
<li>Implement best practices to collect small volume fractions</li>
</ul>
<blockquote>
Presenter: Andrew Leightner (Market Development Manager – Americas Field Marketing, Waters Corporation)
</blockquote>
<blockquote>
Andy Leightner has over 10 years of experience serving the pharma market from contract research organizations. He specialized in method development and validation leveraging LC, GC, and mass spectrometry to meet complex client needs. He supports pharmaceutical and biopharmaceutical discovery and development in the Americas for Waters Corporation. Andy received a bachelors degree in biology from the University of Wisconsin-Madison.
</blockquote>

Waters Corporation: Using the Waters Fraction Manager Analytical (WFMA) to Achieve UPLC Scale Purification

Using the Waters Fraction Manager Analytical (WFMA) to Achieve UPLC Scale Purification

Learn more about analytical scale purification with Waters.

Waters Fraction Manager Analytical UPLC Scale Purification

Introducing midia-PASEF®, a groundbreaking DIA scan mode utilizing overlapping quadrupole windows for comprehensive ion analysis.
In this webinar, you’ll discover how midia-PASEF offers precise precursor m/z determination, automated processing with the MIDIAID pipeline and supports library-free DIA analyses. You’ll also learn how you can rapidly identify over 40 unique peptides per second for de novo sequencing with highly specific MS/MS spectra and discover the real-world applications of this technology within the proteomics field.
Attend this webinar to: 
<ul>
<li>Learn how midia-PASEF® maximizes data quality, enabling fast peptide identification and library-free DIA analyses</li>
<li>Explore the automated MIDIAID pipeline that streamlines processing with peak detection algorithms and machine learning</li>
<li>Discover how you can obtain specific MS/MS spectra for de novo sequencing and seamless searching</li>
<li>See real-world applications: Gain insights into midiaPASEF’s uses in proteomics research, from characterizing the complex immunopeptidome to advancing targeted therapies</li>
</ul>
<blockquote>
Presenter: Dr. Stefan Tenzer (Professor for Immunopeptidomics, University Medical Center, Mainz)
</blockquote>

Bruker Daltonics: Maximizing Information Content in Data-Independent Acquisition Using midia-PASEF

Maximizing Information Content in Data-Independent Acquisition Using midia-PASEF

How midia-PASEF offers precise precursor m/z determination, automated processing with the MIDIAID pipeline and supports library-free DIA analyses.

Data-Independent Acquisition Using midia-PASEF

L-asparaginase (ASNase) is an enzyme-drug that is approved for the treatment of pediatric acute lymphoblastic leukemia (ALL) but is too toxic for adults. To elucidate potentially targetable mechanisms of toxicity with the ultimate goal of decreasing ASNase toxicity and extending its use to adults, we performed multi-omic profiling of the response to ASNase in adult (16-week-old) mice. We administered a range of sub-toxic and toxic doses of ASNase to healthy mice, collected whole blood samples, processed them to plasma, and used a novel sample preparation with just 20 µL mouse plasma as input. We analyzed the extracts using a multi-omic, bioanalytical LC/MS platform capable of measuring 500+ metabolites, 750+ lipids, and 375 peptides.
A Bio-LC was used with three different separations for metabolites, lipids, and peptides. A fast, sensitive, robust, 6495 LC/TQ MS system was used with comprehensive databases for pathway analysis. The results suggested that ASNase induces multi-modal cell death as evidenced by metabolite-, lipid-, and peptide/protein-level changes that occurred in a dose-dependent manner. 
Future studies will test the hypothesis that inhibiting those modes of cell death will decrease ASNase toxicity while preserving anticancer activity. Overall, our findings provide proof-of-concept that multi-omic analysis of drug mechanisms at the protein-, lipid-, and metabolite-levels can unveil toxicity-associated pathways that can be targeted to potentially improve a drug’s therapeutic index.
<blockquote>
Presenter: Ian Edwards (LC/TQ Product Manager, Agilent Technologies, Inc.)
</blockquote>
<blockquote>
Ian holds a Ph.D. 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.
</blockquote>
<blockquote>
Presenter: Philip Lorenzi, Ph.D. (Associate Professor, Department of Bioinformatics and Computational Biology, The University of Texas MD Anderson Cancer Center)
</blockquote>
<blockquote>
I’m an Associate Professor in the Department of Bioinformatics and Computational Biology at The University of Texas MD Anderson Cancer Center. My laboratory focuses on the development of metabolic therapies for the treatment of cancer (with an emphasis on nutrient metabolism), and I’m also Director of the Proteomics Core Facility and Director of the Metabolomics Core Facility. I’ve co-authored 71 peer-reviewed publications. I co-wrote and was Co-Investigator on a successfully completed $4.5M CPRIT Core Facility Award, managed staff and the budget, and oversaw development of bioanalytical services using current equipment and protocols that we have optimized thoroughly. We’ve developed novel mass spectrometry-based methods for: 1) increasing the accuracy of glutamine and glutamate measurements by minimizing in-source conversion to pyroglutamate; 2) normalizing data based on the DNA content of cell- and tissue-based samples; 3) measuring amino acids from as little as 2 µL of mouse plasma or whole blood; 4) detection of over 2,000 annotated metabolites and lipids per biological sample (including just 2 mg of tissue); 5) stable isotope tracer analyses in vitro and in vivo including methods for measuring energy metabolism; 6) determining the extent and position of 13C incorporation into specific amino acids; and 7) diagnosing and correcting batch effects in metabolomic data. Our multidisciplinary team provides full service contract research in a state-of-the art facility.
</blockquote>

Agilent Technologies: Multi-Omic, Mass Spectrometry-Based Elucidation of Mech. of L-asparaginase Toxicity in a Mouse Model

Multi-Omic, Mass Spectrometry-Based Elucidation of Mech. of L-asparaginase Toxicity in a Mouse Model

We analyzed the extracts using a multi-omic, bioanalytical LC/MS platform capable of measuring 500+ metabolites, 750+ lipids, and 375 peptides.

Multi-Omic, Mass Spectrometry-Based Elucidation of Mech

High performance liquid chromatography (HPLC) is a commonly used technique in the pharmaceutical research and manufacturing sectors to determine the concentration and purity of active pharmaceutical ingredients (APIs) or to measure process related residuals and formulation components. In biopharmaceuticals manufacturing or formulation, polysorbates or poloxamers are commonly present either as process related residuals or as formulation components.
The detection and quantitation of these compounds can be a challenge due to their lack of a chromophore which makes traditional UV detection impossible and interference from proteinaceous APIs. This presentation reviews the development and validation strategies of a robust method for the quantitation of poloxamer P188 in biopharmaceuticals. The method uses charged aerosol detection (CAD) on the Thermo Vanquish Flex HPLC system and has suitable sensitivity to quantitate poloxamer P188 down to 3 µg/mL in both AAV and mAb APIs. 
The method is specific in that it is capable of quantitating poloxamer P188 in common formulations and demonstrates non-interference from polysorbate-80 and PEI.
Join our webinar on Tuesday, July 11th at 15:00 CEST (14:00 GMT, 06:00 PDT, 09:00 EDT) to hear about the work of our guest speaker, Adam Punke, from PPD, Part of Thermo Fisher Scientific. In his presentation, he will share his experience of the development and validation strategies of a robust method for the quantitation of poloxamer P188 in biopharmaceuticals.
<h5>Learning points:</h5>
<ul>
<li>What makes charged aerosol detection so powerful</li>
<li>What benefits it offers</li>
<li>What applications are ideal for charged aerosol detection?</li>
</ul>
<h5>Who should attend:</h5>
<ul>
<li>Laboratory managers and directors</li>
<li>Technology leaders</li>
<li>Principal scientists</li>
<li>Laboratory analysts</li>
</ul>
<blockquote>
Presenter: Adam Punke (Guest Speaker, Sr. Research Scientist, PPD, Part of Thermo Fisher Scientific)
</blockquote>
<blockquote>
Presenter: Chris Tuczemskyi (Moderator, Product Marketing Manager, Thermo Fisher Scientific)
</blockquote>
<blockquote>
Chris Tuczemskyi is a chromatography specialist at Thermo Fisher Scientific, based in Cambridge, UK. After spending a number of years building up his experience with chromatography instruments in a water testing laboratory, he joined Thermo Fisher Scientific in 2014 as a chromatography service engineer for Unity Lab Services. Here he developed his passion for supporting customers using HPLC and in 2018 moved into the application specialist team. More recently, in 2022 he made the move to Product Marketing to help ensure we meet and exceed our customers expectation. In 2009, Chris graduated from the University of Essex in Biochemistry (Bsc Hons).
</blockquote>
<blockquote>
Presenter: Ian Acworth (Analytical Instruments Group, Thermo Fisher Scientific)
</blockquote>
<blockquote>
Ian Acworth is part of the marketing team at Thermo Fisher Scientific, based near Boston, Massachusetts. He directed application chemistry development for several HPLC companies while ensuring a strong focus on customer needs and support. He was part of the team that developed and supported the Charged Aerosol Detector. Ian completed his D. Phil in biochemistry at Oxford University in 1986 and completed his postdoctoral studies in neuroscience at MIT in 1989.
</blockquote>
<blockquote>
Presenter: Frank Steiner (Senior Manager of Product Applications, Thermo Fisher Scientific)
</blockquote>
<blockquote>
Frank Steiner is Senior Manager of Product Applications and coordinates HPLC based scientific collaborations in Thermo Fisher Scientific as scientific advisor. He joined Dionex Corporation in 2005, now a part of Thermo Fisher Scientific, and had been manager in various HPLC marketing functions. Frank received his PhD degree in chemistry in 1995 from Saarland University in Saarbruecken, Germany where he became assistant professor in 1997 and associate professor in 2003, after his postdoc at the nuclear research center in Saclay, France in 1996. Frank published more than 30 publications in refereed journals and more than 10 text book chapters on HPLC topics.
</blockquote>
<blockquote>
Presenter: Katherine Lovejoy (Product Application Specialist, Thermo Fisher Scientific)
</blockquote>
<blockquote>
Katherine Lovejoy, Ph.D., has been an application chemist focusing on charged aerosol detection at the Thermo Fisher Scientific HPLC factory site since 2016 and has a Ph.D. in inorganic chemistry from the Massachusetts Institute of Technology. She has experience as a chemist reviewer at the FDA and as a scientist at Los Alamos National Laboratory.
</blockquote>

Thermo Scientific: Quantitation of Poloxamer P188 in Biopharmaceutical Formulations Using Charged Aerosol Detection

Quantitation of Poloxamer P188 in Biopharmaceutical Formulations Using Charged Aerosol Detection

We will share his experience of the development and validation strategies of a robust method for the quantitation of poloxamer P188 in biopharmaceuticals.

Quantitation of Poloxamer P188 in Biopharmaceutical by CAD

How about a 20 minute coffee break along with a bite of Empower? Here you go!
Empower custom fields and Excel in Empower LMS - not a conflict of interest?
Excel Compliance is designed as the solution for customers operating in a regulated environment to use excel spreadsheet with confidence. It provides full audit trail that is compliant to regulatory requirements for electronic records and signatures. 
<ul>
<li>Mitigate the compliance risk of traditional uncontrolled spreadsheets and documents
</li>
<li>Enhanced compliance features such as:
<ul>
<li>authorized access control
</li>
<li>automated version control
</li>
<li>document level and cell by cell audit trails
</li>
</ul>
</li>
<li>Centralize and secure laboratory spreadsheets and documents for use in a regulated environment
</li>
</ul>
<blockquote>
Presenter: Ralf Schröder (Informatics Product Marketing Europe·Waters GesmbH)
</blockquote>

Waters Corporation: Excel and Compliance in Empower LMS

Excel and Compliance in Empower LMS

Empower custom fields and Excel in Empower LMS

For therapeutic antibodies and emerging gene-based medicines, light scattering techniques provide multi-attribute quantification (MAQ) and hence play a critical role in product and process development as well as quality control.
Size-exclusion chromatography with multi-angle light scattering (SEC-MALS) is employed throughout the development cycle of the protein therapeutics for bioanalytical characterization.
In addition, SEC-MALS offers a platform method for quantifying multiple quality attributes of AAV-delivered gene therapy products including titer, payload and aggregate content as well as extended biophysical characterization. In the webinar, we share some case studies using XBridge Premier BEH SEC columns combined with light scattering as a platform approach applied to these different pharmaceutical analytes.
<blockquote>
Presenter: Leo Liu, Ph.D. (Application Scientist, Waters-Wyatt Technology)
</blockquote>

Waters Corporation: Successful SEC-MALS analysis of proteins and AAVs

Successful SEC-MALS analysis of proteins and AAVs

Case studies using XBridge Premier BEH SEC columns combined with light scattering as a platform approach applied to these different pharmaceutical analytes.

In the world of protein formulations, just one poorly behaved excipient can ruin your day… and your dataset. The wrong excipient can promote unfolding or aggregation, so having a big toolbox handy to test how a formulation behaves can save the day. Uncle’s unique combination of fluorescence, SLS, DLS and wide variety of applications means the right tool for the job is always handy – thermal stability, isothermal experiments, kD &amp; B22, G22 and more. Uncle even has a tool for those unruly excipients that throw a wrench in your data by interfering with your protein’s intrinsic fluorescence. Uncle’s full-spectrum fluorescence detection sees fluorescence issues, lets you diagnose what’s going on and helps you solve the problem by changing up your analysis.&nbsp;This talk will look at the many tools Uncle can use to screen formulations for their impact on protein stability, and how Uncle helps you keep an eye out for any nasty surprises that oddball excipients can throw your way.<blockquote>Presenter:&nbsp;Andre Mueller, PhD (Marketing Manager, Biologic Solutions, Unchained Labs)Andre Mueller, PhD, is the Product Manager for Aunty and Honeybun at Unchained Labs, where he supports the development of biologic and gene therapy drugs. He specializes in fluorescence, static and dynamic light scattering, focusing on biophysical characterization of proteins and viral vectors. His research background spans from structural biology to plant physiology, with experience in labs across Germany, Sweden, Denmark, and the USA. Andre earned his PhD at the Carlsberg Laboratory and completed a postdoctoral fellowship at Stanford University.Moderator: Kevin Lance, PhD (Marketing Manager, Analytical Instruments, Unchained Labs)</blockquote>

Unchained Labs: Build up your protein formulation toolbox with Uncle

Build up your protein formulation toolbox with Uncle

Uncle helps spot excipient issues early with tools like fluorescence, SLS, and DLS—keeping your protein formulations stable and your data clean.