ISC 2026 - Day 2

<h4>What if you could compress 179 days of grueling laboratory workflow into just 14 hours?</h4>In this episode of ‪ChromatographyTalk‬, ‪Organomation‬ General Manager David Oliva sits down with analytical chemist Laura Penabad to discuss her groundbreaking new research in biocatalysis and high-throughput screening. Evolving engineered enzymes typically requires checking thousands of variants—a massive bottleneck when traditional Liquid Chromatography-Mass Spectrometry (LC-MS) runs take minutes per sample. Laura reveals how her team built a modular screening platform that shatters this limitation, running samples 128 times faster than standard methods without sacrificing analytical accuracy.Laura breaks down the "orchestra" of technologies making this possible, including coupling nanoliter droplet microfluidics with Cyclic Ion Mobility-Mass Spectrometry (cIM-MS) to separate complex chemical isomers in milliseconds. She also shares the chaotic but triumphant behind-the-scenes stories of her PhD journey—from presenting a "comically bad" early poster at ASMS to recruiting an undergraduate mechanical engineer to build custom automation software from scratch.If you are an early-career researcher, a mass spec enthusiast, or curious about the future of synthetic biology and machine learning in chemistry, this episode is packed with invaluable intuition and advice.<h3>Video Transcription</h3><h4>From Months to Hours: Droplet Ion Mobility–MS for Biocatalysis Screening</h4>High-throughput screening is central to directed enzyme evolution, but conventional analytical workflows can create a difficult compromise: extremely fast screens often provide limited structural information, while techniques such as LC-MS offer much richer chemical insight at the cost of throughput. In a recent interview, analytical chemist Laura described how her work with droplet-based cyclic ion mobility mass spectrometry is helping bridge that gap—providing rapid analysis while retaining the ability to distinguish structurally similar products.Her research grew from an initially modest proof of concept into a screening platform capable of analyzing a 96-well plate in roughly ten minutes. The resulting workflow was approximately 128 times faster than a conventional three-minute LC-MS analysis, opening new possibilities for directed evolution campaigns in which thousands of enzyme variants need to be evaluated.<h4>Directed evolution needs better analytical screening</h4>Biocatalysis takes advantage of enzymes as highly selective catalysts capable of performing reactions under relatively mild conditions. Directed evolution extends those capabilities by deliberately modifying enzyme sequences, expressing large numbers of variants, screening their activity, and using the best-performing mutants as starting points for subsequent rounds of evolution.The process can be repeated multiple times, progressively steering an enzyme toward improved activity, altered substrate specificity, or even chemistry that does not occur naturally.But the effectiveness of directed evolution depends heavily on the screening step. The more variants that can be analyzed—and the more useful chemical information that can be collected for each one—the more effectively researchers can explore the enormous sequence space available to an enzyme.For Laura, this created a clear analytical challenge: develop a screening platform that was substantially faster than conventional chromatographic analysis without sacrificing the structural discrimination needed for chemically similar products.<h4>From an imperfect first experiment to a working platform</h4>The project did not begin with a polished high-throughput workflow.Laura recalled looking back at her 2023 ASMS poster, where the early system was still working with standards rather than true isomeric samples. Droplet introduction was slow—around six seconds per droplet—and the signals were far from the ideal reproducible, square, flat-topped profiles expected from replicate droplets.Nevertheless, simply producing a droplet signal represented an important first milestone.The decisive change came when the project stopped being treated merely as a demonstration that droplet ion mobility–MS could work and instead became a deliberate effort to build a robust screening platform.Laura approached the problem in modules. First came reliable droplet introduction. Then the ion mobility separation method. Quantification became another independent component. Matrix effects were introduced later, gradually making the system resemble the conditions expected with real screening samples.Each layer was tested before another was added.This stepwise strategy became one of the central principles of the project: rather than attempting to optimize the entire system simultaneously, individual components were developed, challenged, and then assembled into an increasingly realistic workflow.<h4>Finding the middle ground between speed and structural information</h4>One of the biggest analytical problems in high-throughput screening is distinguishing products with the same nominal or exact mass.Constitutional isomers may contain the same atoms and therefore generate the same mass-to-charge ratio, while differing only in how functional groups are arranged within the molecular structure. Very rapid optical or mass-based screens may therefore struggle to distinguish them.Traditional LC-MS can solve many of these problems through chromatographic separation, but a several-minute analysis rapidly becomes impractical when thousands of enzyme variants must be screened.Ion mobility offered Laura’s team another option.Rather than separating compounds on a chromatographic timescale, ion mobility can differentiate ions according to their gas-phase behavior on a much shorter timescale. In the workflow described in the interview, this created a useful compromise: not the kilohertz throughput possible with some simple screening technologies, but dramatically faster structural differentiation than conventional LC-MS.The goal was therefore not simply to achieve the fastest possible screen, but to find a middle-throughput analytical regime with enough resolving power to answer chemically meaningful questions.<h4>A 96-well plate in about ten minutes</h4>That strategy eventually produced one of the most striking results of the project.The droplet cyclic ion mobility–MS platform processed a 96-well plate in roughly ten minutes, while maintaining quantitative results comparable with the conventional LC-MS workflow used for comparison. According to the interview, this represented an approximately 128-fold increase in analysis speed relative to a three-minute LC-MS run.Laura described the successful plate experiment less as a surprise than as confirmation that the modular development strategy had worked.Before reaching that point, the workflow had already been challenged through a sequence of increasingly demanding experiments: droplet formation, ion mobility separation, calibration, matrix-containing calibration, and cross-testing analytes at different concentrations. By the time the complete plate was analyzed, each individual part of the analytical system had already been tested under progressively more realistic conditions.<h4>What does that speed mean for directed evolution?</h4>The impact becomes even clearer when the workflow is considered at the scale of an entire directed-evolution campaign.Laura’s manuscript estimated that a five-round campaign involving 5,000 variants would require approximately 179 days using the conventional LC-MS workflow but only about 14 hours using the new platform.Compressing months of analytical work into less than a day has consequences beyond laboratory productivity.It allows researchers to screen more enzyme variants and therefore sample a larger portion of sequence space. But the resulting datasets may also become increasingly valuable for computational approaches.Rather than retaining information only about the best-performing enzyme variant, researchers can preserve data describing variants that performed poorly as well as those that succeeded. When genotype and phenotype information remain linked through the well-plate workflow, these larger datasets can potentially provide richer input for machine-learning models designed to guide subsequent rounds of enzyme evolution.Faster analysis therefore does more than accelerate an existing experiment—it can change the scale and design of the experiment itself.<h4>Balancing ion mobility resolution and signal</h4>Increasing cyclic ion mobility separation time can improve resolution, but it also introduces an important trade-off: additional passes can reduce signal.Laura optimized this balance experimentally by continuously infusing mixtures containing both isomers and progressively adjusting the ion mobility conditions. After tuning parameters such as wave height and velocity, increasing the separation to three passes provided baseline resolution at the high analyte concentrations used during method development.<blockquote>Concentration, however, also mattered.</blockquote>At higher ion populations, space-charge effects could broaden peaks and reduce the baseline separation. This meant that the optimal number of passes could not be selected independently of the concentration range required by the final assay.Her broader advice for optimization is therefore application-driven: decide first what the analytical method actually needs to accomplish.More speed will almost always require sacrificing something—resolution, signal intensity, quantitative precision, or another performance parameter. The key is to identify which compromise the application can tolerate and then optimize the system around that requirement.<h4>Solving droplet carryover with a practical compromise</h4>Droplet-based workflows present their own engineering challenges, including carryover between neighboring samples.In Laura’s system, analytes showed little tendency to partition into the fluorinated oil separating aqueous droplets within the Teflon tubing. The more troublesome carryover occurred closer to electrospray ionization, where a small amount of sample could remain associated with the interface and appear when the next droplet arrived.Rather than adding excessive engineering complexity to an already elaborate platform, the researchers adopted a pragmatic solution: a sacrificial wash droplet between analytical samples.It is an example of a recurring theme throughout the project. The objective was not to construct the theoretically most sophisticated droplet system, but to create a screening workflow that remained sufficiently robust, reproducible, and practical for large numbers of samples.<h4>High-throughput analysis also requires high-throughput data processing</h4>Accelerating the mass spectrometry experiment created another bottleneck: the data.The instrument produced a continuous intensity-versus-time trace rather than a neatly organized well-plate dataset. With small experiments, Laura could manually identify individual droplets and process the results in spreadsheets. That approach quickly became unrealistic as sample numbers increased.The solution emerged through collaboration with an undergraduate mentee, Aiden, who helped develop a dedicated software tool for processing the continuous traces.Laura defined how the software should behave and how the analytical information should be organized, while Aiden developed the computational implementation. The workflow itself was subsequently adjusted to make automated data processing more reliable—for example, by introducing marker columns that allowed the software to recognize the end of each plate row.The resulting tool can accept the acquired traces together with information about plate dimensions and replicates and automatically return parameters including peak identity, duration, and the corresponding well position. A later version also generated an automatically populated heat map.The experience illustrates how improving analytical throughput often requires simultaneous development of instrumentation, experimental design, and informatics. Making the measurement faster is useful only if the resulting data can be processed at a comparable speed.<h4>Mentoring became part of the scientific process</h4>The software project also reflects another theme Laura emphasized throughout the interview: the value of mentoring.Several younger researchers contributed directly to the development of the final platform. Laura described mentees not simply as additional hands in the laboratory but as scientists capable of proposing their own hypotheses and experimental ideas.Working with undergraduate researchers also influenced the way she organized experiments.Because their available laboratory time was limited, each task needed to have a clear purpose. That encouraged a more disciplined approach to experimental planning: instead of performing experiments simply because they could be done, the team had to ask which experiment would most efficiently answer the next important question.In that sense, mentoring became another mechanism for improving the research itself.<h4>The next step: continuous, automated screening</h4>Despite the dramatic increase in analytical speed, the current workflow still includes manual operations such as loading tubing and connecting the sample stream to the mass spectrometer.The next engineering challenge is therefore automation.Laura described ongoing work using the Venturi effect to pull samples directly from wells into the droplet system. Building on previous work in the laboratory with 96-well plates, she has already tested this approach with as many as 2,000 samples.The long-term direction is clear: continuous and robust droplet generation, automated handling of thousands of samples, rapid ion mobility separation, mass spectrometric detection, and automated reconstruction of the resulting data into plate-based outputs.Combining those elements could move the system closer to the type of large-scale screening required for increasingly ambitious biocatalysis campaigns.<h4>Learning mass spectrometry one problem at a time</h4>Laura also reflected on the broader experience of developing the platform during her PhD.Ion mobility and mass spectrometry initially appeared intimidating, particularly because they combine concepts from analytical chemistry, physical chemistry, separations, instrumentation, and electrical engineering. What eventually changed was not simply learning more facts, but developing the ability to move repeatedly between theory and experiment.An unexpected result sends the researcher back to the underlying theory. A better understanding suggests a modified experiment. That experiment creates another observation, gradually building intuition about how the system behaves.Her advice to early-career scientists is therefore to accept that this process takes time.Advanced analytical techniques may initially appear overwhelming, but understanding develops incrementally through repeated interaction between theory, experimentation, failure, and refinement. Equally important, she emphasized the role of the supportive ion mobility–mass spectrometry community in making a technically demanding field more approachable for young researchers.From the first inconsistent droplets to a platform capable of processing thousands of samples, that iterative mindset ultimately shaped the entire project.<blockquote>This text has been automatically transcribed from a video presentation using AI technology. It may contain inaccuracies and is not guaranteed to be 100% correct.</blockquote><blockquote><h4>Concentrating on Chromatography Podcast</h4>Dive into the frontiers of chromatography, mass spectrometry, and sample preparation with host David Oliva. Each episode features candid conversations with leading researchers, industry innovators, and passionate scientists who are shaping the future of analytical chemistry. From decoding PFAS detection challenges to exploring the latest in AI-assisted liquid chromatography, this show uncovers practical workflows, sustainability breakthroughs, and the real-world impact of separation science. Whether you’re a chromatographer, lab professional, or researcher you'll discover inspiring content!You can find Concentrating on Chromatography Podcast in podcast apps:<ul><li><a target="_blank" rel="noopener noreferrer" href="https://open.spotify.com/show/1wZg67rCjoGju4yhqxI3tL">Spotify</a></li><li><a target="_blank" rel="noopener noreferrer" href="https://podcasts.apple.com/tz/podcast/concentrating-on-chromatography/id1788429185">Apple podcast</a></li><li><a target="_blank" rel="noopener noreferrer" href="https://podscan.fm/podcasts/concentrating-on-chromatography">podscan</a></li></ul>and on <a target="_blank" rel="noopener noreferrer" href="https://www.youtube.com/@ChromatographyTalk/videos">YouTube channel</a></blockquote>

Droplet Cyclic Ion Mobility Mass Spec: Compressing Months of Enzyme Screening Into Just 14 Hours

ISC 2026 - Day 3

Article

Events

Video

Scientists

Interview

The International Symposium on Chromatography (ISC) represents the oldest conference series focussing on separation science. ISC symposia have been organised since 1956 in each even year. ISC is internationally recognised as one of the premier meeting series for discussion of all modes of chromatography and separation science with a broad coverage of techniques and applications.

International Symposium on Chromatography - (ISC)

Organomation, founded in 1959, designs and manufactures high-quality nitrogen evaporators and extraction systems. Known for innovation and durability, their lab instruments are used globally for efficient sample preparation with strong customer support.

Organomation

KNAUER Eurospher II 100 C18A (U)HPLC Columns (USP L1)

C18A phases can be operated under 100 % aqueous conditions without destroying the stationary phase, what is not recommended for classical C18 phases

C18A columns for polar reversed phase applications with 100 % aqueous mobile phase (USP L1)C18 stationary phase for the analysis of more polar and hydrophilic compounds in reversed phase mode (USP L1). Silica gel based columns with C18 modification with hydrophilic endcapping. Because of their modification, C18A phases can be operated under 100 % aqueous conditions without destroying the stationary phase, what is not recommended for classical C18 phases caused by their hydrophobicity.<h5>Properties</h5><ul><li>Separation mechanism: Hydrophobic and polar interaction</li><li>Ultra pure, spherical high performance HPLC phase based on silica gel</li><li>Polar, monomeric C18 (Octadecyl) modification, polar endcapping, with 10 % carbon content (~ 50 % endcapping).</li></ul><h5>Key features</h5><ul><li>C18 A phase with alternative, more polar selectivity, suited for the use with pure aqueous mobile phases, 50 % endcapping and resulting 10 % carbon content, 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>Polar endcapped C18 phase for water soluble and polar analytes, 100 % aqueous eluents for analysis of very polar compounds:<ul><li>basic pharmaceutical ingredients</li><li>water soluble vitamins</li><li>catecholamines</li><li>as well as organic acids.</li></ul></li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_C18_A_6f4b0c3063.webp" alt="KNAUER: C18A" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_C18_A_6f4b0c3063.webp 245w," sizes="100vw" width="690" height="428"><blockquote>Tip: With an endcapping the influence of nonderivatized silanol groups can be minimized. The endcapping reagent is generally a smaller silane than used for derivatisation. This treatment reduces the unwanted interaction of polar or charged analytes (acids, bases ect.) because the amount of available silanol groups is reduced.</blockquote>Stationary phase Eurospher II 100 C18A is provided in these column types: analytical, semi-preparative and preparative.Eurospher II 100 C18A 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 C18A 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 Eurospher II 100 C18H (U)HPLC Columns (USP L1)

C18H stationary phase with a special, high efficiency or polymeric unpolar endcapping style. Suited for the use of mobile phase in an extended pH range from typically pH 1–12. 

Hydrophobic / pH stable C18H Columns for more unpolar analytes in Reversed Phase (USP L1)C18H stationary phase with a special, high efficiency or polymeric unpolar endcapping style. Because of the high carbon content and the protection of the Silica gel these materials are also suited for the use of mobile phase in an extended pH range from typically pH 1–12.While a high endcapped C18H phase is characterized mostly by the high carbon content. A polymeric endcapped phase also provides an increased steric selectivity.<h5>Properties</h5><ul><li>Separation mechanism: Hydrophobic interaction</li><li>Ultra pure, spherical high performance HPLC phase based on silica gel</li><li>Nonpolar, monomeric C18 (Octadecyl) modification, high efficiency endcapping with 17 % carbon content (~ 99 % endcapping)</li></ul><h5>Key features</h5><ul><li>Nonpolar C18 phase with high efficiency endcapping and 17 % carbon content, resulting in higher pH stability in the range of 1–12, 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>Recommended alternative to Kromasil 100 C18, for acidic, basic and neutral analytes in reversed phase mode with extended pH range.</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_C18_H_c5531f7d87.webp" alt="KNAUER: C18H" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_C18_H_c5531f7d87.webp 226w," sizes="100vw" width="687" height="475"><blockquote>Tip: All analytical KNAUER columns can be used in reversed flow direction for example for cleaning. Please refer to our Column care and use documents on the website for further information about storage conditions and cleaning procedures.</blockquote>Stationary phase Eurospher II 100 C18H is provided in these column types: analytical, semi-preparative and preparative.Eurospher II 100 C18H 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 C18H 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.<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">

KNAUER Eurospher II 100 C18P (U)HPLC Columns (USP L1)

KNAUER´s most unpolar C18P phase with excellent shape selectivity. It features polymeric, high efficiency endcapping of 99 % and 20 % carbon content, exceptional pH stability in the range of 1–12.

C18P columns with trifunctional modifications (USP L1): Hydrophobic and stearic interactionsKNAUER´s most unpolar C18P phase with excellent shape selectivity. It features polymeric, high efficiency endcapping of 99 % and 20 % carbon content, exceptional pH stability in the range of 1–12, and outstanding mechanical and chemical stability.<h5>Properties</h5><ul><li>Separation mechanism: Hydrophobic and stearic interaction</li><li>Ultra pure, spherical high performance HPLC phase based on silica gel</li><li>Unpolar, trifunctional C18 (Octadecyl) modification, high efficiency endcapping, with 20 % carbon content (~ 99 % endcapping)&nbsp;</li></ul><h5>Key features</h5><ul><li>KNAUER´s most unpolar C18 phase, polymeric, high efficiency endcapping of 99 % and 20 % carbon content, higher pH stability in the range of 1-12, 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>Alternative selectivity to standard C18; stationary phase in Eurospher II C18 family with the highest carbon load; fully endcapped; excellent shape selectivity and pH stability.</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_C18_P_f093126fa9.webp" alt="KNAUER: C18P" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_C18_P_f093126fa9.webp 245w," sizes="100vw" width="278" height="174"><blockquote>Tip: Not all C18 phases are comparable! When replacing an existing column, always have a close look at the characteristics like carbon load, pore size and specific surface area.</blockquote>Stationary phase Eurospher II 100 C18P is provided in these column types: analytical, semi-preparative and preparative.Eurospher II 100 C18P 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 C18P 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 Eurospher II 100 C4 (U)HPLC Columns (USP L26)

C4 stationary phases (USP L26) are often used in reversed phase mode for different application areas. 

C4 columns for Reversed Phase in HPLC and UHPLC (USP L26)C4 stationary phases (USP L26) are often used in reversed phase mode for different application areas.&nbsp;They are suitable for:<ul><li>Hydrophobic Interaction Chromatography (HIC mode)</li><li>Analysis of biomolecules like proteins and peptides</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_C4_3f3adfbdb4.webp" alt="KNAUER: C4" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_C4_3f3adfbdb4.webp 78w," sizes="100vw" width="200" height="400"><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.<img 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">

KNAUER Eurospher II 100 CN (U)HPLC Columns for NP & RP (USP L10)

CN is one of the most versatile stationary phases that can be used in reversed phase mode as well as in normal phase mode. 

Cyano or CN for Normal Phase and Reversed Phase (USP L10)CN is one of the most versatile stationary phases that can be used in reversed phase mode as well as in normal phase mode. The cyano modification supports the π-π interaction as well as polar and hydrophobic interactions with low polarity solvents. These phases are preferred for compounds with double bonds and aromatic groups. Cyano bonded phases show unique selectivity for polar compounds and are more suitable than bare silica for normal phase (NP) gradient separations. The cyano functional group is a strong dipole that can interact with other dipoles or induce dipoles on solutes. These phases also exhibit moderate hydrophobicity due to the alkyl linker.Due to the quick equilibration time of the CN bonded phase, it is the best choice for gradient elution in normal phase mode. CN phases can be used as a less polar alternative to Si phases.The material is typically preferred for:<ul><li>Compounds with double bonds</li><li>Cyclic antidepressants</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_CN_4fff7a1e28.webp" alt="KNAUER: CN" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_CN_4fff7a1e28.webp 130w," sizes="100vw" width="358" height="428"><blockquote>Stationary phase Eurospher II 100 CN is provided in these column types: analytical, semi-preparative and preparative.</blockquote>Eurospher II 100 CN Analytical Columns are available with these parameters:<ul><li>Column length (mm): 50, 100, 150, 250</li><li>Column ID (mm): 3, 4, 4.6</li><li>Particle size (µm): 3, 5</li></ul>Eurospher II 100 CN 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.<img 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">

KNAUER Eurospher II 100 DNPH (U)HPLC Columns (USP L1)

Silica gel based columns with C18 modification with special endcapping, designed for DNPH analysis.

DNPH columns for polar reversed phase applications with 100 % aqueous mobile phase (USP L1)C18 stationary phase for the analysis of more polar and hydrophilic compounds in reversed phase mode (USP L1). Silica gel based columns with C18 modification with special endcapping, designed for DNPH analysis. Because of their modification, DNPH phases can be operated under 100 % aqueous conditions without destroying the stationary phase, what is not recommended for classical C18 phases caused by their hydrophobicity.These phases are ideal for the analysis of:<ul><li>DNPH-derivates of aldehydes and ketones</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_DNPH_7020417932.webp" alt="KNAUER: DNPH" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_DNPH_7020417932.webp 245w," sizes="100vw" width="690" height="428"><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.<img 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">

KNAUER Eurospher II 100 Diol (U)HPLC Columns for NP (USP L20)

Diol packing material can be universally used in different polar modes (NP, RP and HILIC). Diol bonded phases are a versatile alternative to unbonded silica for normal phase separations.

Diol columns for Normal Phase in HPLC and UHPLC (USP L20)Diol packing material can be universally used in different polar modes (NP, RP and HILIC). Diol bonded phases are a versatile alternative to unbonded silica for normal phase separations with shorter equilibration times.The hydroxyl groups provide good selectivity without excessive retention, caused by the weaker H-bonding of the diol layer than of silanols. Some diol bonded phases have been developed specifically for applications in the HILIC mode.<h5>Properties</h5><ul><li>Separation mechanism: Hydrophilic interaction</li><li>Ultra pure, spherical high performance HPLC phase based on silica gel</li><li>Diol, no endcapping, with 5 % carbon content&nbsp;</li></ul><h5>Key features</h5><ul><li>High-class HPLC phase perfectly suited to take on routine analyses as well as the most ambitious chromatography tasks, classical Diol phase without endcapping and 5 % carbon content, outstanding mechanical and chemical stability.</li></ul><h5>Recommended application areas</h5><ul><li>Alternative to the silica packing with shorter equilibration time and comparable selectivity; due to the lower activity of these packings it can be also used for SEC-applications.</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Diol_26fe64c2b9.webp" alt="KNAUER: Diol" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Diol_26fe64c2b9.webp 156w," sizes="100vw" width="426" height="425"><blockquote>Tip: The most important column parameters for evaluation are retention time, peak symmetry and theoretical plates. Retention time to prove the reproducibility of measurements. Peak symmetry to estimate how good the column bed is packed (peak symmetry value of 1.0 is optimal). Theoretical plate numbers to determine the effectiveness of the HPLC column.</blockquote>Stationary phase Eurospher II 100 Diol is provided in these column types: analytical, semi-preparative and preparative.Eurospher II 100 Diol Analytical Columns are available with these parameters:<ul><li>Column length (mm): 50, 100, 150, 250</li><li>Column ID (mm): 3, 4, 4.6</li><li>Particle size (µm): 3, 5</li></ul>Eurospher II 100 Diol 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.<img 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">

KNAUER Eurospher II 100 Phenyl (U)HPLC Columns for RP (USP L11)

Phenyl stationary phases (USP L11) in reversed phase mode with alternative selectivity for aromatic and moderately π-π interaction polar analytes

Phenyl columns for Reversed Phase in HPLC and UHPLC &nbsp;(USP L11)Phenyl stationary phases (USP L11) in reversed phase mode with alternative selectivity for:<ul><li>Aromatic and moderately π-π interaction polar analytes</li><li>Mixtures with varying polarity and aromaticity</li></ul>The polarity is similar to C8 modification but with different selectivity for polar aromatics, polycyclic aromatic hydrocarbons, fatty acids etc. Phenyl phases with a classical slightly hydrophobic endcapping or without endcapping.<h5>Properties</h5><ul><li>Separation mechanism: π-π interactions</li><li>Ultra pure, spherical high performance HPLC phase based on silica gel</li><li>Phenyl modification (Phenylpropyl), 12 % carbon content</li></ul><h5>Key features</h5><ul><li>Classical phenyl phase with 12 % 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>Alternative selectivity for aromatic and mode rately pi-pi interaction polar analytes or mixtures with varying polarity and aromaticity.</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Phenyl_df9b20c77a.webp" alt="KNAUER: Phenyl" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Phenyl_df9b20c77a.webp 90w," sizes="100vw" width="306" height="528"><blockquote>Tip: For UHPLC columns like Eurospher II 2 μm particle size, samples and eluents should be filtered through a 0.2 μm instead of a 0.45 μm membrane filter. Frits at column inand outlet have smaller pores than classical HPLC columns and block more easily.</blockquote>Stationary phase Eurospher II 100 Phenyl is provided in these column types: analytical, semi-preparative and preparative.Eurospher II 100 Phenyl 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 Phenyl 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.<img 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">

KNAUER Eurospher II 100 HILIC (U)HPLC Columns

Based on ultra-pure Silica gel, zwitterionic HILIC phases are ideal for the analysis of polar and ionizable compounds. Stronger interaction of more polar compounds with stationary aqueous layer.

Zwitterionic and polar columns for HILIC applicationsBased on ultra-pure Silica gel, zwitterionic HILIC phases are ideal for the analysis of polar and ionizable compounds. Stronger interaction of more polar compounds with stationary aqueous layer.Faster elution of nonpolar compounds is possible due to minor hydrophobic interactions. Retention increases with the charge of the analyte.<h5>Recommended for the analysis of hydrophilic compounds like:</h5><ul><li>Polar organic acids and bases</li><li>Natural polar compounds</li><li>Nucleosides</li><li>Oligonucleotides</li><li>Amino acids</li><li>Peptides</li><li>Water soluble vitamins</li></ul><h5>Properties</h5><ul><li>Separation mechanism: Hydrophilic and weak electrostatic interactions</li><li>Ultra pure, spherical high performance HPLC phase based on silica gel</li><li>Zwitterionic modification: Ammonium – sulfonic acid</li><li>7 % carbon content</li></ul><h5>Key features</h5><ul><li>Modern HILIC phase with zwitterionic modification on the basis of ammonium- sulfonic acid. Outstanding mechanical and chemical stability.</li></ul><h5>Recommended application areas</h5><ul><li>Especially suited for the separation of hydrophilic, polar and ionic analytes which are poorly retained on reversed phase columns; behavior is the other way round on Eurospher II HILIC compared to RP which makes it an ideal tool to enhance chromatographic separations for these molecules.</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_HILIC_b9c3bffc20.webp" alt="KNAUER: HILIC" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_HILIC_b9c3bffc20.webp 147w," sizes="100vw" width="439" height="465"><blockquote>Tip: HILIC applications are well-suited for coupling with MS detection. The high organic content of the applied mobile phases are qualified well for evaporation in the MS´s ionization source.</blockquote>Stationary phase Eurospher II 100 HILIC is provided in these column types: analyticalEurospher II 100 HILIC Analytical Columns are available with these parameters:<ul><li>Column length (mm): 50, 100, 150, 250</li><li>Column ID (mm): 3, 4, 4.6</li><li>Particle size (µm): 3, 5</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.<img 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">

KNAUER Eurospher II 100 NH2 (U)HPLC Columns for NP & HILIC (USP L8)

Amino (NH2) bonded silica gel phases can be used in the normal phase (NP) and HILIC mode, but also for reversed phase (RP) mode.

Amino columns for Normal Phase and for HILIC applications (USP L8)Amino (NH2) bonded silica gel phases can be used in the normal phase (NP) and HILIC mode, but also for reversed phase (RP) mode. They show alternative normal phase selectivity to unbonded silica, especially for aromatics. Furthermore, the selectivity is influenced by hydrogen bond interactions with the analyte. Amino columns can also be used in the HILIC mode for carbohydrate analysis and for other polar compounds. Their weak anion-exchange properties can be used in the analysis of anions and organic acids.The amino group is bonded to the silica support via a propyl linker.<h5>Properties</h5><ul><li>Separation mechanism: Hydrophilic and ionic interactions</li><li>Ultra pure, spherical high performance HPLC phase based on silica gel</li><li>Amino modification (Aminopropyl)</li><li>4 % carbon content (without endcapping)</li></ul><h5>Key features</h5><ul><li>Multi mode column for RP, NP, HILIC and IC, Aminopropyl modification without endcapping, outstanding mechanical and chemical stability.</li></ul><h5>Recommended application areas</h5><ul><li>Most flexible phase in the Eurospher II family; can be used in three modes: normal phase, reversed phase and ion chromatography mode (weak anion exchanger); different selectivity to the silica packing; in reversed phase mode mainly used for analysis of carbohydrates.&nbsp;</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_NH_2_HILIC_NP_fbaa6027c0.webp" alt="KNAUER: NH2" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_NH_2_HILIC_NP_fbaa6027c0.webp 107w," sizes="100vw" width="290" height="421"><blockquote>Tip: A precolumn can help to protect your analytical column. Especially when working with highly matrix afflicted samples it is highly recommended to use precolumns.</blockquote>Stationary phase Eurospher II 100 NH2 is provided in these column types: analytical, semi-preparative and preparative.Eurospher II 100 NH2 Analytical Columns are available with these parameters:<ul><li>Column length (mm): 50, 100, 150, 250</li><li>Column ID (mm): 3, 4, 4.6</li><li>Particle size (µm): 3, 5</li></ul>Eurospher II 100 NH2 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.<img 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">

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