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.<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"><h3>Why PFAS analysis matters</h3>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.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.<h3>Addressing the challenges of PFAS identification</h3>Successful non-targeted PFAS analysis requires overcoming several analytical hurdles:<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><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1759">Thermo Scientific Compound Discoverer 3.5 software </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">HRAM Orbitrap MS</a> data with an expanded suite of PFAS-specific libraries and databases.In a single workflow, users can:<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">mzCloud</a>, NIST 2023, and in silico 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>With these resources combined, the software enables confident annotation of more than 40,000 PFAS compounds!<h3>New tools for confident, streamlined PFAS identification</h3>Compound Discoverer software introduces several new features designed to simplify and accelerate PFAS NTA workflows:<ul><li>Apply PFAS Scoring Node: Automatically calculates PFAS-specific parameters and a new PFAS Score—no R scripting required.</li><li>PFAS Plot: A visual tool that uses mass-defect metrics to highlight probable PFAS in complex datasets, following methods from Kaufman et al. (2022).</li><li>Expanded suite of libraries and databases: Combines mzCloud, NIST, in silico, PFAS fragment databases, and a new neutral loss database for greater confidence.</li><li>Advanced data visualization: Tools like Kendrick Mass Defect and Molecular Network plots reveal homologous PFAS series and related compound families.</li><li>Built-in statistical analysis tools: Compare PFAS composition between samples using Principal Component Analysis, Differential analysis and Hierarchical Cluster Analysis.</li></ul><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"><h3>From complexity to clarity</h3>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.<a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/5612">Watch the on-demand webinar</a> to see the full workflow in action and learn how Compound Discoverer software can enhance your PFAS analysis.<ul><li>Visit us on&nbsp;<a target="_blank" rel="noopener noreferrer" href="https://www.linkedin.com/showcase/72003132/admin/dashboard/">LinkedIn:</a>&nbsp;#PFAS #CompoundDiscoverer #SpectralLibraries</li></ul><blockquote><h5>Zoe Julian</h5>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.</blockquote>

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

<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

Software

Article

Product

Application

Academy

Video

A unique company dedicated to serving science helping our customers drive productivity and accelerate results. Through our premier brands – Thermo Scientific, Applied Biosystems, Invitrogen, Fisher Scientific and Unity Lab Services.

Thermo Fisher Scientific

Everything from the world of analytical chemistry in one place. We connect people in solving their problems. At Labrulez you will find all the necessary information easily, quickly and clearly. Stop searching and start finding.

LabRulez

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

KNAUER

KNAUER Eurospher II 100 C8 & C8A (U)HPLC Columns (USP L7)

C8 is a silica gel based stationary phase in reversed phase mode with alternative selectivity compared to C18 phases. 

Classical and hydrophilic/aqueous C8 columns for HPLC and UHPLC (USP L7)C8 is a silica gel based stationary phase in reversed phase mode with alternative selectivity compared to C18 phases. Compared to C18 phases, these columns show less retention due to the lower hydrophobicity.<ul><li>Ultra pure, spherical high performance HPLC phase based on silica gel</li><li>Monomeric C8 (Octyl) modification, standard endcapping, with 10 % carbon content (~ 50 % endcapping)</li></ul><h5>Properties</h5><ul><li>Separation mechanism: Hydrophobic interaction(lower compared to C18 phases)</li></ul><h5>Key features</h5><ul><li>Classical C8 phase, standard 50 % endcapping and 10 % carbon load, outstanding mechanical and chemical stability, suited for analytical as well as semi preparative and preparative applications.</li></ul><h5>Recommended application areas</h5><ul><li>Similar selectivity to C18 phase but less retention due to the lower hydrophobicity; useful for determination of:<ul><li>Water soluble vitamins</li><li>Steroids</li><li>Catecholamines</li><li>Sedatives</li></ul></li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_C8_rp_non_endcapped_3f7727d3fd.webp" alt="KNAUER: C8 RP Non-endcapped" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_C8_rp_non_endcapped_3f7727d3fd.webp 148w," sizes="100vw" width="304" height="321">Hydrophilic C8A phase offers an alternative, more polar selectivity. It is suited for highly polar reversed phase applications with up to 100 % aqueous mobile phases.&nbsp;<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_C8_A_e1f371165e.webp" alt="KNAUER: C8A" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_C8_A_e1f371165e.webp 198w," sizes="100vw" width="473" height="373"><blockquote>Tip: Caution! Even for short term storage all buffer solutions have to be rinsed from the column to prevent crystallization effects.</blockquote>Stationary phases Eurospher II 100 C8 and C8A are provided in these column types: analytical, semi-preparative and preparative.Eurospher II 100 C8 and C8A Analytical Columns are available with these parameters:<ul><li>Column length (mm): 50, 100, 150, 250</li><li>Column ID (mm): 2, 3, 4, 4.6</li><li>Particle size (µm): 2, 3, 5</li></ul>Eurospher II 100 C8 and C8A Semi-Preparative Standard Columns are available with these parameters:<ul><li>Column length (mm): 30, 50, 125, 150, 250</li><li>Column ID (mm): 8, 16</li><li>Particle size (µm): 5, 10</li></ul><h3>KNAUER (U)HPLC Columns</h3>Widest range of HPLC and UHPLC columns for all separation modes and applications.<ul><li>Safe Investment: Robust and reliable columns for attractive price&nbsp;</li><li>Separation Mode: Reversed Phase, HILIC, Normale</li><li>Simplicity: Easy purchase via our online store with fast delivery</li><li>Application: All-rounder for research or highly specialised for high-throughput analysis? We have the best column for your requirements.</li><li>Flexibility: Wide selection of column and particle sizes, for (U)HPLC applications up to 1000 bar</li></ul>KNAUER offers the perfect column for any small molecule separation. No matter which separation mode, molecule and the mobile phase, we offer a suitable (U)HPLC stationary phase. Choose from a wide range of modifications, particle sizes and column dimensions to suit your application. To elongate the lifetime of your column, you can choose to use a suitable precolumn.<figure class="image"><img style="aspect-ratio:787/674;" src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_KNAUER_U_HPLC_C8_and_C8_A_Columns_083c60f401.webp" alt="KNAUER: KNAUER (U)HPLC Columns" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_KNAUER_U_HPLC_C8_and_C8_A_Columns_083c60f401.webp 182w," sizes="100vw" width="787" height="674"></figure>

KNAUER Eurosil Bioselect 300 C8 HPLC Columns (USP L7)

Eurosil Bioselect C8 has been specifically developed for the determination and purification of peptides, proteins, and oligonucleotides up to a molecular weight of approximately 20 kDa.

Analytical wide pore phase for the determination of biomolecules bigger than 5000 DaEurosil Bioselect C8 has been specifically developed for the determination and purification of peptides, proteins, and oligonucleotides up to a molecular weight of approximately 20 kDa.<h5>Properties</h5><ul><li>Ultra pure, spherical wide pore HPLC phase based on wide pore silica gel</li><li>Unpolar, monomeric C8 modification, endcapped</li><li>4.5 % carbon content (conventional endcapping)</li><li>Separation mechanism: hydrophobic interaction</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_C8_hydrosphob_6986c292b4.webp" alt="KNAUER: C8" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_C8_hydrosphob_6986c292b4.webp 144w," sizes="100vw" width="381" height="413"><h5>Recommended application areas</h5><ul><li>Hydrophilic biomolecules, peptides, peptide mapping after enzymatic digestion</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Eurosil_Bioselect_Phases_98acc32d0e.jpg" alt="KNAUER: Eurosil Bioselect - Phases and recommended sizes of molecules" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Eurosil_Bioselect_Phases_98acc32d0e.jpg 245w," sizes="100vw" width="1385" height="778"><h3>KNAUER Wide pore HPLC columns</h3>For enhanced resolution, capacity and recovery of proteins and other biomolecules<ul><li>Material: Silica or polymer particles</li><li>Separation Ranges: 2–100 kDa</li><li>Column Brands: Eurosil Bioselect or AppliChrom®</li><li>Particle sizes: 3–10 µm</li><li>Pore Sizes: 300 or 100–1000 Å</li><li>Specific surface: 90 ± 5 m2/g</li><li>Pore volume: 0.8 ml/g</li><li>Density: 450 g/l</li><li>Applications: Peptides exceeding approximately 50 amino acids and oligonucleotides greater than 25 residues</li><li>Customize: Bioanalysis requires a lot of flexibility in the pore size, the pH range, the temperatures and the solvents used. To achieve this, we cross phase limits.</li></ul><h4>Characteristics&nbsp;</h4><ul><li>Eurosil Bioselect columns are available in 4 modifications, useful for a wide range of separation tasks involving large molecules up to 100 000 Da.</li><li>Strict manufacturing procedures ensure a very narrow pore size distribution.</li><li>Interfering anions or heavy metal ions on the silica surface are not present in Eurosil Bioselect columns because only ultra pure silica gel and high quality chemical modifications are employed.</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Eurosil_Bioselect_85cfc2d201.jpg" alt="KNAUER: Eurosil Bioselect" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Eurosil_Bioselect_85cfc2d201.jpg 222w," sizes="100vw" width="1114" height="782"><h4>Separation mechanism of Eurosil Bioselect</h4>To separate larger molecules, they need to freely access the interior of the pores of the packing material. Therefore, the analytes’ diameter must be smaller than the average pore diameter.&nbsp;For higher molecular weight solutes, the use of lower pore size materials (60-120 Å) as typically used for the analysis of small molecules may result restricted diffusion in the material´s pores and therewith reduced column efficiency.&nbsp;<h4>Modifications</h4>The recommended modification of Eurosil Bioselect depends on the molecular weight of the analytes.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Eurosil_Bioselect_chemical_modifications_1fd5d87f7b.jpg" alt="KNAUER: Eurosil Bioselect - chemical modifications" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Eurosil_Bioselect_chemical_modifications_1fd5d87f7b.jpg 245w," sizes="100vw" width="955" height="505"> &nbsp;

KNAUER Eurospher II 100 C8 & C8A Prep HPLC Columns (USP L7)

Classical and hydrophilic/aqueous C8 columns for HPLC and UHPLC (USP L7)C8 is a silica gel based stationary phase in reversed phase mode with alternative selectivity compared to C18 phases. Compared to C18 phases, these columns show less retention due to the lower hydrophobicity.<ul><li>Ultra pure, spherical high performance HPLC phase based on silica gel</li><li>Monomeric C8 (Octyl) modification, standard endcapping, with 10 % carbon content (~ 50 % endcapping)</li></ul><h5>Properties</h5><ul><li>Separation mechanism: Hydrophobic interaction(lower compared to C18 phases)</li></ul><h5>Key features</h5><ul><li>Classical C8 phase, standard 50 % endcapping and 10 % carbon load, outstanding mechanical and chemical stability, suited for analytical as well as semi preparative and preparative applications.</li></ul><h5>Recommended application areas</h5><ul><li>Similar selectivity to C18 phase but less retention due to the lower hydrophobicity; useful for determination of:<ul><li>Water soluble vitamins</li><li>Steroids</li><li>Catecholamines</li><li>Sedatives</li></ul></li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_C8_rp_non_endcapped_3f7727d3fd.webp" alt="KNAUER: C8 RP Non-endcapped" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_C8_rp_non_endcapped_3f7727d3fd.webp 148w," sizes="100vw" width="304" height="321">Hydrophilic C8A phase offers an alternative, more polar selectivity. It is suited for highly polar reversed phase applications with up to 100 % aqueous mobile phases.&nbsp;<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_C8_A_e1f371165e.webp" alt="KNAUER: C8A" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_C8_A_e1f371165e.webp 198w," sizes="100vw" width="473" height="373"><blockquote>Tip: Caution! Even for short term storage all buffer solutions have to be rinsed from the column to prevent crystallization effects.</blockquote>Stationary phases Eurospher II 100 C8 and C8A are provided in these column types: analytical, semi-preparative and preparative.Eurospher II 100 C8 and C8A Preparative Standard Columns are available with these parameters:<ul><li>Column length (mm): 50, 150, 250</li><li>Column ID (mm): 20, 30, 50</li><li>Particle size (µm): 5, 10, 15</li></ul>Eurospher II 100 C8 and C8A Preparative dAX Columns with dynamic axial compression are available with these parameters:<ul><li>Column length (mm): 50, 150, 250</li><li>Column ID (mm): 25, 30, 50</li><li>Particle size (µm): 5, 10, 15</li></ul><h3>KNAUER Preparative HPLC Columns</h3>The right choice for all dimensions from semi-prep to batch chromatography to SMB.<ul><li>Modifications: Reversed Phase, HILIC, Normale Phase</li><li>Max. Pressure: 400–600 bar</li><li>Column Inner Diameter (ID): 16–50 mm in our shop, more on request</li><li>Column Brands: Eurospher, Eurospher II</li><li>Safe Investment: Robustness and durability based on superior technology</li><li>Application: Easy upscaling, as all materials are available from ultra high performance to preparative scale HPLC. You can also use our upscaling calculator.</li><li>Customizable: Standard flanged hardware &amp; dAX hardware with static or dynamic axial compression available</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Preparative_HPLC_Columns_31bc2477c3.webp" alt="KNAUER: KNAUER Preparative HPLC Columns" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Preparative_HPLC_Columns_31bc2477c3.webp 221w," sizes="100vw" width="1024" height="724"><h5>Preparative HPLC columns can be offered with two different hardware types:</h5><ul><li>Versatile standard flanged hardware for nearly any application task in batch chromatography</li><li>dAX Hardware with static and dynamic axial compression for an extended lifetime and multi-column chromatography like SMB processes</li></ul>

Products from category C8 HPLC columns focused on LC columns distributed by KNAUER - page 1

Related content

News from LabRulezLCMS Library - Week 14, 2026

Oligos Made Easy - Part 9: 2D-LC

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