<p><strong>Per- and polyfluoroalkyl substances (PFAS) have become a major global concern due to their persistence, bioaccumulation, and potential health impacts. With thousands of known PFAS compounds, and new variants continuing to emerge, identifying and quantifying them across complex environmental and biological samples is an ongoing analytical challenge.</strong></p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Thermo_Fisher_Scientific_From_Complexity_to_Clarity_Smarter_PFAS_Identification_Workflows_in_Compound_Discoverer_Software_2_8e83167891.jpg" alt="Thermo Fisher Scientific: From Complexity to Clarity: Smarter PFAS Identification Workflows in Compound Discoverer Software" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Thermo_Fisher_Scientific_From_Complexity_to_Clarity_Smarter_PFAS_Identification_Workflows_in_Compound_Discoverer_Software_2_8e83167891.jpg 245w," sizes="100vw" width="1024" height="574"></p><h3><strong>Why PFAS analysis matters</strong></h3><p>PFAS are used in everything from firefighting foams to non-stick coatings, yet their chemical stability means they persist in the environment and accumulate in living organisms. Regulators and scientists around the world are racing to better understand their distribution, fate, and toxicity.</p><p>But with tens of thousands of PFAS, many without available reference standards, traditional targeted analysis alone isn’t enough. Analysts increasingly rely on non-targeted analysis (NTA) workflows powered by high-resolution accurate-mass (HRAM) mass spectrometry and advanced software to uncover the full picture.</p><h3><strong>Addressing the challenges of PFAS identification</strong></h3><p>Successful non-targeted PFAS analysis requires overcoming several analytical hurdles:</p><ol><li>Acquiring high-quality MS¹ and MS² data with ultra-high mass resolution and accuracy.</li><li>Detecting low-abundance PFAS hidden within thousands of background compounds.</li><li>Leveraging comprehensive spectral libraries and databases for confident identification.</li><li>Applying data filtering and scoring tools to assign confidence levels to annotations.</li><li>Using visualization and statistical tools to interpret and compare complex datasets.</li></ol><p><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1759"><strong>Thermo Scientific Compound Discoverer 3.5 software </strong></a>brings together all the components needed for robust PFAS identification—integrating <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products?category=22&amp;manufacturerItems=3"><strong>HRAM Orbitrap MS</strong></a> data with an expanded suite of PFAS-specific libraries and databases.</p><p><strong>In a single workflow, users can:</strong></p><ul><li>Detect and annotate compounds automatically from LC-MS data.</li><li>Determine possible elemental formulas using isotope pattern and accurate mass information.</li><li>Compare MS² spectra against <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1764"><strong>mzCloud</strong></a>, NIST 2023, and <i>in silico</i> PFAS libraries.</li><li>Cross-reference annotations with the EPA PFAS Structure List and the NIST Suspect List of PFAS.</li><li>Use PFAS-specific fragment and neutral loss databases to strengthen identification confidence.</li></ul><p>With these resources combined, the software enables confident annotation of more than<strong> 40,000 PFAS </strong>compounds!</p><h3><strong>New tools for confident, streamlined PFAS identification</strong></h3><p>Compound Discoverer software introduces several new features designed to simplify and accelerate PFAS NTA workflows:</p><ul><li><strong>Apply PFAS Scoring Node:</strong> Automatically calculates PFAS-specific parameters and a new PFAS Score—no R scripting required.</li><li><strong>PFAS Plot:</strong> A visual tool that uses mass-defect metrics to highlight probable PFAS in complex datasets, following methods from Kaufman et al. (2022).</li><li><strong>Expanded suite of libraries and databases:</strong> Combines mzCloud, NIST, <i>in silico</i>, PFAS fragment databases, and a new neutral loss database for greater confidence.</li><li><strong>Advanced data visualization:</strong> Tools like Kendrick Mass Defect and Molecular Network plots reveal homologous PFAS series and related compound families.</li><li><strong>Built-in statistical analysis tools</strong>: Compare PFAS composition between samples using Principal Component Analysis, Differential analysis and Hierarchical Cluster Analysis.</li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Thermo_Fisher_Scientific_From_Complexity_to_Clarity_Smarter_PFAS_Identification_Workflows_in_Compound_Discoverer_Software_ed32c7f7c1.jpg" alt="Thermo Fisher Scientific From Complexity to Clarity Smarter PFAS Identification Workflows in Compound Discoverer Software" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Thermo_Fisher_Scientific_From_Complexity_to_Clarity_Smarter_PFAS_Identification_Workflows_in_Compound_Discoverer_Software_ed32c7f7c1.jpg 245w," sizes="100vw" width="800" height="395"></p><h3><strong>From complexity to clarity</strong></h3><p>By uniting Orbitrap HRAM data with the most comprehensive PFAS libraries available, Compound Discoverer 3.5 software streamlines non-targeted PFAS analysis—helping laboratories move from data complexity to confident chemical clarity.</p><p><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/5612"><strong>Watch the on-demand webinar</strong></a> to see the full workflow in action and learn how Compound Discoverer software can enhance your PFAS analysis.</p><ul><li><strong>Visit us on&nbsp;</strong><a target="_blank" rel="noopener noreferrer" href="https://www.linkedin.com/showcase/72003132/admin/dashboard/"><strong><u>LinkedIn:</u></strong></a>&nbsp;#PFAS #CompoundDiscoverer #SpectralLibraries</li></ul><blockquote><h5><strong>Zoe Julian</strong></h5><p>Zoe Julian is a Product Marketing Manager for Chromatography and Mass Spectrometry software at Thermo Fisher Scientific. With over 12 years' experience of working with chromatography and mass spectrometry techniques with a background in GC and GC-MS(/MS) organic environmental analysis, she has helped many laboratories and industries improve their workflows through a unique understanding of their challenges. Transitioning to a focus on Chromeleon CDS she has shown customers how software solutions can improve their productivity and support their day-to-day operations.</p></blockquote>

From Complexity to Clarity: Smarter PFAS Identification Workflows in Compound Discoverer Software

<h3><strong>Two Dimensions, One Solution: Advancing Oligonucleotide Analysis with 2D-LC</strong></h3><h4><strong>Why Consider 2D-LC for Oligonucleotide Analysis?</strong></h4><p>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.</p><p><strong>Two-dimensional liquid chromatography (2D-LC) </strong>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.</p><blockquote><h4><strong>Previous parts from the Oligos Made Easy</strong> <strong>series</strong></h4><ul><li><strong>Part 1: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/article/6808"><strong>Overview</strong></a></li><li><strong>Part 2: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/article/6846"><strong>AEX</strong></a></li><li><strong>Part 3: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/article/6880"><strong>IP-RP</strong></a></li><li><strong>Part 4: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/article/6912"><strong>HILIC</strong></a></li><li><strong>Part 5: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/article/6944"><strong>SEC</strong></a></li><li><strong>Part 6: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/article/6982"><strong>Column Navigator</strong></a></li><li><strong>Part 7: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/article/7012"><strong>Large Scale AEX</strong></a></li><li><strong>Part 8: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/article/7045"><strong>Sepapure oliGO</strong></a></li></ul></blockquote><h4><strong>Moving Beyond the Limits of 1D-LC</strong></h4><p>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.</p><p><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"></p><p>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.</p><p>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.</p><h4><strong>Architecture of a 2D-LC System</strong></h4><p>At its core, a 2D-LC system relies on precise coordination between its components. The <strong>setup typically consists</strong> of <strong>two chromatographic columns</strong>, <strong>two pumping systems</strong>, and a <strong>switching valve</strong> 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.</p><p><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"></p><p>The <strong>valve plays a crucial role</strong> in this process, acting as the interface between both chromatographic dimensions. Its function is to alternately <strong>collect fractions from the first dimension and inject them into the second dimension</strong> 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: <strong>while one loop is being filled, the other is being flushed into the second dimension</strong>.</p><p><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"></p><p>This alternating process requires <strong>precise timing</strong>. 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.</p><p><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"></p><h4><strong>Applying 2D-LC to Oligonucleotide Analysis</strong></h4><p>In oligonucleotide workflows, 2D-LC combines two complementary separation strategies to address sample complexity. The <strong>first dimension</strong> is typically used for <strong>bulk separation</strong> based on properties such as <strong>size, charge, or polarity</strong>. Techniques such as <strong>size-exclusion chromatography</strong> (SEC), <strong>anion-exchange chromatography</strong> (AEX), <strong>ion-pair reversed-phase chromatography </strong>(IP-RP), <strong>hydrophilic interaction chromatography</strong> (HILIC), or <strong>mixed-mode chromatography</strong> (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.</p><p>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.</p><p>A <strong>key advantage of 2D-LC</strong> lies in the <strong>flexibility of combining different stationary phases and mobile phase</strong> conditions.&nbsp;</p><p>Numerous separation combinations are possible, including:</p><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><p>This adaptability allows tailored workflows for specific analytical challenges.</p><h4><strong>Heart-Cutting vs. Comprehensive 2D-LC</strong></h4><p>When implementing 2D-LC, two primary transfer strategies are commonly used: <strong>heart-cutting</strong> and <strong>comprehensive transfer</strong>. Each approach offers distinct advantages depending on the analytical objective.</p><p><strong>Heart-cutting 2D-LC</strong> 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.</p><p><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"></p><p>In contrast, <strong>comprehensive 2D-LC</strong> 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.</p><p><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"></p><p>In practice, <strong>hybrid approaches may also be employed</strong>, combining targeted heart-cutting with broader comprehensive analysis to balance resolution and coverage.</p><h4><strong>Practical Considerations for Method Development</strong></h4><p>Successful implementation of 2D-LC for oligonucleotide analysis requires careful method design.&nbsp;</p><h5><strong>1. Column selection</strong>&nbsp;</h5><p>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.</p><h5><strong>2. Mobile phase selection&nbsp;</strong></h5><p>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.</p><h5><strong>3. Instrumentation&nbsp;</strong></h5><p>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.</p><h5><strong>4. Method development&nbsp;</strong></h5><p>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.</p><h4><strong>Final Remarks</strong></h4><p>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.</p><p>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.</p><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"><strong>sales@knauer.net</strong></a>. Stay tuned for more exciting insights into the Oligonucleotide world in our “<strong>Oligos Made Easy</strong>” series.</li><li>For more in-depth discussion or questions, reach out to the author at <a href="kindler@knauer.net"><strong>kindler@knauer.net</strong></a></li></ul></blockquote>

Oligos Made Easy - Part 9: 2D-LC

News from LabRulezLCMS Library - Week 14, 2026

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Waters ACQUITY APC Columns

Waters ACQUITY Advanced Polymer Chromatography (APC) Columns with novel capabilities for chromatographic efficiency, speed and resolution that cannot be matched by conventional GPC methods.

<h4><strong>ACQUITY APC AQ BEH Column</strong></h4><p>ACQUITY APC AQ columns are designed to provide highly efficient size-based separations for water soluble polymer characterization. The sorbent uses patented Ethylene Bridge Hybrid (BEH) technology to provide a mechanical strong packed bed that resists shinking and swelling for easy mobile phase solvent switching. The expected molecular weight range for this column is approximatley 1, 000 - 30, 000 g/mole.</p><h4><strong>Specifications</strong></h4><ul><li><strong>Separation Mode:</strong> SEC/GPC</li><li><strong>Particle Substrate: </strong>Hybrid</li><li><strong>pH Range Min:</strong> 1 pH</li><li><strong>pH Range Max: </strong>8 pH</li><li><strong>Endcapped: </strong>No</li><li><strong>Silanol Activity: </strong>Medium</li><li><strong>Molecular Weight Range Min:</strong> 1000</li><li><strong>Molecular Weight Range Max:</strong> 30000</li><li><strong>Particle Shape: </strong>Spherical</li><li><strong>Particle Size:</strong> 1,7 µm &nbsp;/ 2.5 µm&nbsp;</li><li><strong>Endfitting Type: </strong>Parker-style</li><li><strong>System:</strong> APC</li><li><strong>Particle Technology:</strong> BEH</li><li><strong>Technique: </strong>SEC</li></ul><h4><strong>ACQUITY APC XT BEH Column</strong></h4><p>ACQUITY APC XT columns are optimized to provide highly efficient size-based separations for organic solvent soluble polymer characterization. The sorbent uses patented Ethylene Bridge Hybrid (BEH) technology to provide a mechanical strong packed bed that resists shinking and swelling for easy mobile phase solvent switching.The expected molecular weight range for this column is approximatley 1,000 - 30,000 g/mole.</p><h4><strong>Specifications</strong></h4><ul><li><strong>Separation Mode:</strong> SEC/GPC</li><li><strong>Particle Substrate: </strong>Hybrid</li><li><strong>pH Range Min:</strong> 1 pH</li><li><strong>pH Range Max: </strong>12 pH</li><li><strong>Endcapped: </strong>No</li><li><strong>Silanol Activity: </strong>Low</li><li><strong>Molecular Weight Range Min:</strong> 1000</li><li><strong>Molecular Weight Range Max:</strong> 30000</li><li><strong>Particle Shape: </strong>Spherical</li><li><strong>Particle Size:</strong> 1,7 µm &nbsp;/ 2.5 µm&nbsp;</li><li><strong>Endfitting Type: </strong>Parker-style</li><li><strong>System:</strong> APC</li><li><strong>Particle Technology:</strong> BEH</li><li><strong>Technique: </strong>GPC</li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_ACQUITY_APC_AQ_BEH_Column_0c0d837b73.jpeg" alt="Waters: ACQUITY APC Column" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_ACQUITY_APC_AQ_BEH_Column_0c0d837b73.jpeg 234w," sizes="100vw" width="660" height="440"></p><h3><strong>ACQUITY APC Columns</strong></h3><h4><strong>Achieve superior speed and resolution for polymer characterization</strong></h4><p>Achieving superior performance and reliability with high resolution polymer characterization can be a challenge. <strong>ACQUITY Advanced Polymer Chromatography (APC) Columns</strong> are designed for polymer scientists like you, with novel capabilities for chromatographic efficiency, speed and resolution that cannot be matched by conventional GPC methods. With these columns, you can quantify and characterize polymer samples with confidence and accuracy while maximizing productivity.</p><p>The <strong>Ethylene Bridged Hybrid (BEH) particles</strong> used in ACQUITY APC Columns enable rapid solvent switching and the ability to use multiple conditions for the same bank of columns. Conventional polymeric stationary phases are dedicated to a single set of analytical conditions and solvent because the chromatographic bed is vulnerable to shrinking and swelling. By eliminating this undesirable behavior, ACQUITY APC Columns allow for maximum robustness and method development flexibility.</p><h4><strong>Overview</strong></h4><ul><li>Eliminate the shrinking and swelling that often occurs with polymeric stationary phases upon solvent switching with a stable chromatographic column bed&nbsp;</li><li>Achieve unmatched separation speed, resolution, and chromatographic efficiency from optimized pore volumes and particle size</li><li>Meet workflow requirements with 1.7 and 2.5 μm columns for organic and aqueous size-based separation</li><li>Achieve industry-leading column-to-column reproducibility with exceptionally reliable columns from a tightly controlled column manufacturing process</li><li><strong>Recommended Use: </strong>For rapid and accurate chromatographic characterization of synthetic polymer and macromolecular species.&nbsp;</li></ul><h4><strong>ACQUITY APC Column Featured Technologies</strong></h4><h5><strong>BEH Technology</strong></h5><ul><li>Rugged hybrid organic/inorganic base particle</li><li>Improved peak shape for bases- low silanol activity</li><li>Wider operating pH range for method development flexibility</li><li>Mechanical strength for maximum column efficiency at UHPLC operating pressures</li></ul><h4><strong>ACQUITY APC Column Characteristics</strong></h4><h5><strong>ACQUITY APC XT Columns</strong></h5><ul><li>Designed to provide high efficiency sized-based separations for organic soluble polymer characterization</li><li>Uses BEH particle technology</li><li>Stable column bed that resists shrinking and swelling for easy mobile-phase solvent switching</li><li>Features an extended temperature range up to 90 °C</li><li>Designed for organic solvent-based separations</li></ul><h5><strong>ACQUITY APC AQ Columns</strong></h5><ul><li>Designed to provide high efficiency sized-based separations for water soluble polymer characterization</li><li>Uses BEH particle technology</li><li>Robust separations up to 45 °C</li><li>Designed for aqueous-soluble polymer separations</li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_ACQUITY_APC_Columns_2268ceb5f5.jpeg" alt="Waters: ACQUITY APC Columns" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_ACQUITY_APC_Columns_2268ceb5f5.jpeg 234w," sizes="100vw" width="1024" height="682"></p><h4><strong>Unprecedented capacity for rapid solvent switching</strong></h4><p>Use multiple conditions for the same column bank with rapid solvent switching using the rigid hybrid particles in ACQUITY APC Chromatography Columns. Conventional polymeric phases are often dedicated to single analysis conditions because the chromatographic bed is vulnerable to shrinking and swelling that can be potentially harmful to the column. By eliminating this constraint, ACQUITY APC Chromatography Columns provide more freedom for method development and deliver advanced polymer characterization for a diverse range of samples.</p><h4><strong>Reduce calibration time by a factor of 3-5x and increase resolution</strong></h4><p>With long conventional Gel Permeation Chromatography (GPC) run times of up to one hour or more, analyses of standards and samples can be quite time-consuming. In addition to long GPC run times, the large extra-column volume of conventional GPC systems can result in peak bandspreading, reducing the resolution and thus accuracy of the calibration points. Combined with the lower dispersion ACQUITY APC System, small particle ACQUITY APC Columns deliver less bandspreading and narrower, sharper peaks compared to the conventional GPC system. Additionally, robust sub-3-µm APC Column Technology supports higher flow rate and backpressures, thereby improving resolution.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_A_comparison_of_run_times_and_resolution_for_polystyrene_standards_Mp_100_K_10_K_and_1_K_on_a_conventional_GPC_system_and_the_ACQUITY_APC_System_37cfb15ed5.jpeg" alt="Waters: A comparison of run times and resolution for polystyrene standards (Mp: 100K, 10K, and 1K) on a conventional GPC system and the ACQUITY APC System. " srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_A_comparison_of_run_times_and_resolution_for_polystyrene_standards_Mp_100_K_10_K_and_1_K_on_a_conventional_GPC_system_and_the_ACQUITY_APC_System_37cfb15ed5.jpeg 234w," sizes="100vw" width="1024" height="682"></p><h4><strong>Ensure reproducible column performance</strong></h4><p>Reproducible column performance is one of the most critical factors for achieving long-term method reliability during polymer characterization. Each step of our column manufacturing process is monitored and tightly controlled to maintain unmatched product reproducibility. Every test, from raw material characterization to final batch analysis, is designed to ensure that ACQUITY APC Columns perform the same from column-to-column, batch-to-batch, and year-to-year.</p>

Waters SEC Columns

Get unmatched batch-to-batch reproducibility with the wide range of Waters Size-Exclusion Chromatography (SEC) Columns and standards for protein and peptide characterization.

<h4><strong>Size-based separations of peptides, proteins, nucleic acids, LNPs, and viral vectors</strong></h4><p>Get unmatched batch-to-batch reproducibility with the wide range of <strong>Waters Size-Exclusion Chromatography (SEC) Columns</strong> and standards for protein and peptide characterization.</p><p><strong>SEC</strong> is a powerful technique for aggregate and fragment analysis in the <strong>research, development</strong>, and <strong>manufacturing of biotherapeutic proteins</strong>, such as <strong>insulin</strong> and <strong>monoclonal antibodies</strong> (mAb). With SEC, also known as gel filtration, you can separate proteins based on their size (hydrodynamic radii versus absolute molecular weight) in solution, with larger sized species eluting before smaller proteins.</p><p>The <strong>pore size and volume</strong> of the SEC particles used in the packed column drives this <strong>isocratic based technique</strong>, so the column you choose matters. As the molecular weight of the proteins of interest increases, SEC columns with comparatively larger pore size particles are selected.</p><h4><strong>Overview</strong></h4><ul><li>Gain efficiency with this “platform based” method for <strong>proteins ranging from 10,000 to 650,000 Daltons</strong>, leveraging <strong>MaxPeak Premier Technology</strong> coupled with <strong>BEH-PEO Particle Technology&nbsp;</strong> &nbsp;&nbsp;</li><li><strong>Minimize non-desired secondary interactions </strong>for improved separation reproducibility and results with Ethylene Bridged Hybrid (BEH) SEC particles</li><li>Ensure reliable <strong>column-to-column and batch-to-batch performance</strong> with columns quality control tested with relevant peptide, protein, and mAb samples &nbsp;</li><li>Achieve reliable <strong>high throughput analysis on UPLC</strong> systems with our <strong>1.7 µm particle size</strong> offering and reproducible separations on any <strong>HPLC, UHPLC, or UPLC system</strong> to address different application needs with <strong>2.5 µm&nbsp;</strong></li><li><strong>Extend analytical column life</strong> and<strong> lower cost per analyses </strong>with SEC Guards&nbsp;</li><li>Confirm on-site SEC column and LC system performance with available BEH125, BEH200, BEH450, and mAb Size Variant Standards &nbsp;&nbsp;</li><li><strong>Manufactured in a cGMP, ISO 9001</strong> certified plant using stringent manufacturing protocols and ultra-pure reagents</li></ul><p><strong>Recommended Use: </strong>For size-based peptide, protein, and viral vector analyses.</p><h4><strong>System Dispersion Specifications for Column Recommendations</strong></h4><h5><strong>LC System Dispersions &lt;20 µL (UPLC)</strong></h5><ul><li>ACQUITY Premier Protein SEC 250 Å Guards and Columns</li><li>ACQUITY UPLC Protein BEH SEC 125 Å, 200 Å, and 450 Å Guards and Columns</li><li>1.7 µm guards and columns for low dispersion UPLC systems</li></ul><h5><strong>LC System Dispersions &gt;20 µL – ≤35 µL (UHPLC)</strong></h5><ul><li>XBridge Premier Protein SEC 250 Å Guards and Columns</li><li>XBridge Protein BEH SEC 125 Å, 200 Å, and 450 Å Guards and Columns</li><li>BioResolve mAb SEC Guards and Columns</li><li>2.5 µm guards and columns for UHPLC systems</li></ul><h5><strong>LC System Dispersions &gt;35 µL (HPLC)</strong></h5><ul><li>XBridge Protein BEH SEC 125 Å, 200 Å, and 450 Å Guards and Columns</li><li>XBridge Premier Protein SEC 250 Å Guards and Columns</li><li>BioResolve mAb SEC Guards and Columns</li><li>BioSuite SEC Guards and Columns</li><li>3.5 µm guards and columns for HPLC systems</li></ul><h4><strong>GTxResolve SEC Columns</strong></h4><h5><strong>GTxResolve Premier BEH SEC 450 Å Columns</strong></h5><ul><li>2.5 µm particle size columns for UHPLC and UPLC separations</li><li>Lower secondary interactions and lower noise suitable for platform methods</li><li>Low adsorption hardware necessary for analysis of biologics samples</li><li>450 Å pore size with up to 50% higher resolving power reducing need for sample consumption, resulting in faster analyses<ul><li><a target="_blank" rel="noopener noreferrer" href="https://www.waters.com/nextgen/us/en/products/columns/GTxResolve-sec-columns.html"><strong>Explore GTxResolve SEC Columns</strong></a></li></ul></li></ul><h5><strong>GTxResolve Premier SEC 1000 Å Columns</strong></h5><ul><li>3 µm columns for UHPLC and UPLC separations</li><li>1000 Å columns with high efficiency for aggregate analysis of GTx drugs with diameter of 80-200 nm</li><li>Low secondary interaction &amp; improved compatibility with universally applied detector, such as MALS</li><li>Low adsorption hardware necessary for analysis of biologics samples<ul><li><a target="_blank" rel="noopener noreferrer" href="https://www.waters.com/nextgen/us/en/products/columns/GTxResolve-sec-columns.html"><strong>Explore GTxResolve SEC Columns</strong></a></li></ul></li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_G_Tx_Resolve_SEC_Columns_1b8741a62b.jpg" alt="Waters: GTxResolve SEC Columns" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_G_Tx_Resolve_SEC_Columns_1b8741a62b.jpg 234w," sizes="100vw" width="1024" height="682"></p><h4><strong>Protein SEC Columns</strong></h4><h5><strong>ACQUITY Premier Protein SEC 250 Å Guards and Columns</strong></h5><ul><li>Sub-2-μm particle columns for UPLC separations</li><li>250 Å pore size</li><li>High throughput &amp; resolution SEC separations of biomolecules ranging from 10,000 to 650,000 Daltons on low dispersion UPLC Systems</li><li>Available SEC Guard Columns to extend analytical SEC Column life</li><li>Significantly reduce non-desired ionic and hydrophobic secondary interaction for desired SEC platform methods with MaxPeak Premier Technology and BEH-PEO SEC particle technology<ul><li><a target="_blank" rel="noopener noreferrer" href="https://www.waters.com/nextgen/us/en/products/columns/acquity-premier-columns.html"><strong>Explore ACQUITY Premier Protein BEH SEC Guards and Columns</strong></a></li></ul></li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_ACQUITY_Premier_Columns_featuring_Max_Peak_HPS_Technology_11a541350e.jpg" alt="Waters: ACQUITY Premier Columns featuring MaxPeak HPS Technology" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_ACQUITY_Premier_Columns_featuring_Max_Peak_HPS_Technology_11a541350e.jpg 234w," sizes="100vw" width="1024" height="682"></p><h5><strong>XBridge Premier Protein SEC 250 Å Guards and Columns</strong></h5><ul><li>2.5 μm particle columns for HPLC and UHPLC separations</li><li>250 Å pore size</li><li>High resolution SEC separations of biomolecules ranging from 10,000 to 650,000 Daltons on any LC system</li><li>Available SEC Guard Columns to extend analytical SEC Column life</li><li>Significantly reduce non-desired ionic and hydrophobic secondary interaction for desired SEC platform methods with MaxPeak Premier Technology and BEH-PEO SEC particle technology</li><li>Specifically designed and QC tested with NIST-based, mAb Size Variant Standard to help ensure batch-to-batch consistency<ul><li><a target="_blank" rel="noopener noreferrer" href="https://www.waters.com/waters/global/Waters-XBridge-Premier-Columns/nav.htm"><strong>Explore XBridge Premier Protein BEH SEC Columns</strong></a></li></ul></li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_X_Bridge_Premier_Columns_0230c90278.jpg" alt="Waters: XBridge Premier Columns" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_X_Bridge_Premier_Columns_0230c90278.jpg 193w," sizes="100vw" width="950" height="768"></p><h5><strong>BioResolve mAb SEC Guards and Columns</strong></h5><ul><li>2.5 μm particle columns for UPLC, UHPLC, and HPLC separations</li><li>200 Å pore size</li><li>Obtain reproducible, accurate quantitation of monoclonal antibodies (mAbs), associated high molecular weight aggregates (&gt;300,000 Da), and lower molecular weight fragments (e.g., &lt;100,000 Da)</li><li>Each packed column is tested with NIST-based, mAb size variant standard to help ensure batch-to-batch and column-to-column performance consistency<ul><li><a target="_blank" rel="noopener noreferrer" href="https://www.waters.com/waters/global/BioResolve-SEC-mAb-Consumables-%7C-SEC-Guards/nav.htm"><strong>Explore BioResolve mAb SEC Guards and Columns</strong></a></li></ul></li></ul><h5><strong>ACQUITY UPLC Protein BEH SEC Guards and Columns</strong></h5><ul><li>Sub-2-μm particle columns for UPLC separations (2.5 μm for 450 Å)</li><li>Pore size options include: 125 Å, 200 Å, and 450 Å</li><li>Available SEC Guard Columns to extend analytical SEC Column life</li><li>Specifically designed and QC tested with BEH125, BEH200, or BEH450 protein standards to help ensure batch-to-batch consistency</li><li>10x faster than traditional HPLC-based SEC<ul><li><a target="_blank" rel="noopener noreferrer" href="https://www.waters.com/nextgen/us/en/products/columns/acquity-uplc-columns.html"><strong>Explore ACQUITY UPLC Protein BEH SEC Guards and Columns</strong></a></li></ul></li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_Acquity_UPLC_kolony_1dc5c25d3e/Waters-Acquity-UPLC-kolony.jpg" alt="Waters: Acquity UPLC columns" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_Acquity_UPLC_kolony_1dc5c25d3e/thumbnail_Waters-Acquity-UPLC-kolony.jpg 245w," sizes="100vw" width="807" height="407"></p><h5><strong>XBridge Protein BEH SEC Columns</strong></h5><ul><li>2.5 μm particle columns for use on any UPLC, UHPLC, or HPLC system</li><li>Pore size options include: 125 Å, 200 Å, and 450 Å</li><li>Available SEC Guard Columns to extend analytical SEC Column life</li><li>Specifically designed and QC tested with BEH125, BEH200, or BEH450 protein standards to help ensure batch-to-batch consistency<ul><li><a target="_blank" rel="noopener noreferrer" href="https://www.waters.com/waters/global/XBridge-BEH-Columns-for-High-pH-Chromatography/nav.htm"><strong>Explore XBridge Protein BEH SEC Columns</strong></a></li></ul></li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_X_Bridge_Columns_b0b55de501/Waters-XBridge-Columns.jpg" alt="Waters: XBridge Columns" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_X_Bridge_Columns_b0b55de501/thumbnail_Waters-XBridge-Columns.jpg 203w," sizes="100vw" width="900" height="691"></p><h4><strong>Peptide SEC Columns</strong></h4><h5><strong>BioSuite SEC Columns</strong></h5><ul><li>Silica-based diol SEC columns</li><li>Molecular weight exclusion limits are determined primarily by the selected pore size</li><li>Available in analytical and semi-preparative column sizes<ul><li><a target="_blank" rel="noopener noreferrer" href="https://www.waters.com/waters/global/BioSuite-Columns/nav.htm"><strong>Explore BioSuite SEC Columns</strong></a></li></ul></li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_Bio_Suite_Columns_bf2bb8249f.jpg" alt="Waters: BioSuite Columns" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_Bio_Suite_Columns_bf2bb8249f.jpg 184w," sizes="100vw" width="307" height="260"></p><h5><strong>Protein-Pak SEC Columns</strong></h5><ul><li>Based on a 10 μm diol-bonded silica</li><li>Resolve proteins that differ in molecular weight by a factor of two</li><li>Distinguish proteins differing by as little as 15% in molecular weight<ul><li><a target="_blank" rel="noopener noreferrer" href="https://www.waters.com/waters/global/Protein-Pak-HPLC-and-UPLC-Columns/nav.htm"><strong>Explore Protein-Pak SEC Columns</strong></a></li></ul></li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_Protein_Pak_Columns_be6a669326.jpg" alt="Waters: Protein-Pak Columns" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_Protein_Pak_Columns_be6a669326.jpg 245w," sizes="100vw" width="500" height="315"></p><h4><strong>Choose the right SEC Column for your lab</strong></h4><p>SEC is ideal for aggregate and fragment analysis in the research, development, and manufacturing of biotherapeutic peptides, proteins, and viral vectors, but choosing the right column for your analysis is important. Use the following criteria to choose a column that provides reliable, high quality SEC analyses of various biomolecules, including insulin, monoclonal antibodies, and adeno associated viruses (AAVs).</p><h5><strong>What to consider when choosing an SEC Column:</strong></h5><ul><li>What is the molecular weight of your sample?</li><li>What is the dispersion of your LC system?</li><li>Are you separating components &lt;2 fold difference in molecular weight (MW)?</li><li>Are you separating components &gt;2 fold difference in MW?</li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_Choose_the_right_Size_exclusion_chromatography_SEC_column_for_your_analysis_with_Waters_SEC_columns_selection_guide_b81f674f65.jpg" alt="Waters: Choose the right Size-exclusion chromatography (SEC) column for your analysis with Waters SEC columns selection guide." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_Choose_the_right_Size_exclusion_chromatography_SEC_column_for_your_analysis_with_Waters_SEC_columns_selection_guide_b81f674f65.jpg 158w," sizes="100vw" width="1024" height="1013"></p><h4><strong>Addressing the complexity of SEC Separations</strong></h4><p>Experience the advancements in SEC-based technologies to address non-desired specific ionic and hydrophobic interactions for more reliable results with ACQUITY Premier Protein SEC and XBridge Premier Protein SEC Columns and Guards.</p><h5><strong>With MaxPeak Premier SEC Columns, you can:</strong></h5><ul><li>Reproducibly resolve injected sample components that are less than 2x different in size for challenging applications</li><li>Use a ‘platform SEC method’ to reduce method development time and cost</li><li>Ensure batch-to-batch and column-to-column performance</li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_Classes_of_therapeutic_proteins_separated_on_an_ACQUITY_Premier_Protein_SEC_Column_d3f74a8ea4.jpg" alt="Waters: Classes of therapeutic proteins separated on an ACQUITY Premier Protein SEC Column." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_Classes_of_therapeutic_proteins_separated_on_an_ACQUITY_Premier_Protein_SEC_Column_d3f74a8ea4.jpg 234w," sizes="100vw" width="1024" height="682"></p>

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