<div><p><strong>Understanding how to read the resulting data of high-performance liquid chromatography (HPLC) helps turn raw detector output into confident decisions about purity, identity, and assay results. When you know how to read HPLC peaks and baselines, you can quickly spot problems, verify method performance, and support regulatory‑ready reports.</strong></p></div><div><div><div><div><h3><strong>What is an HPLC Chromatogram?</strong></h3></div></div></div></div><div><p>An HPLC chromatogram is a plot of detector signal versus time that shows how each compound in your sample moves through the column and reaches the detector. Every peak corresponds to a different component, with the position on the time axis giving its retention time and the peak size relating to concentration.</p><p>In routine work, you read retention time to confirm identity and rely on peak area or height for concentration, so learning how to read an HPLC chromatogram is essential for everyday assay workflows.</p></div><div><div><div><div><h3><strong>Why Is It Important to Read HPLC Chromatograms Correctly?</strong></h3><p>Reading HPLC results correctly matters because retention time and peak response are key parameters for separating, identifying, and quantifying compounds in complex mixtures. Misreading the baseline or peak shape can distort area integration, which affects assay accuracy and impurity quantitation.</p><p>Good <a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/resources/knowledge-center/hplc-knowledge-center/what-are-chromatograms"><strong>chromatogram</strong></a> interpretation also helps you detect method drift early, keep system suitability within limits, and avoid costly repeat injections or failed batches.</p></div></div></div></div><div><div><div><div><h3><strong>Key Components of an HPLC Chromatogram</strong></h3></div></div></div></div><div><p>Before you interpret a run, you must recognize the fundamental parts of a chromatogram. Each visual element gives you specific data about the separation and the analytes present in your sample.</p><h4><strong>Baseline</strong></h4><p>The baseline is the detector signal when no analyte peak is eluting, and a stable, flat baseline makes integration and quantification reliable. More detailed descriptions of baseline noise and drift are covered in the troubleshooting section below. In a nutshell, it is enough to note that any instability can affect how confidently you integrate and compare peaks.</p><h4><strong>Peaks</strong></h4><p>Peaks represent eluting analytes and appear as Gaussian‑like shapes rising above the baseline. Symmetric peaks with clear separation are easier to integrate and give more reliable responses for quantitative work.</p><h4><strong>Retention Time</strong></h4><p>Retention time, <i>t</i><sub>R</sub>, is the time from injection until the apex of a peak, reflecting how long an analyte spends in the column compared with the mobile phase. The solvent front or void peak corresponds to the dead time, <i>t</i><sub>0</sub>, which is the passage of unretained material through the system.</p><h4><strong>Peak Area and Height</strong></h4><p>Peak area is generally more reliable than peak height for quantitative analysis, because it better reflects the total amount of analyte passing the detector. In validated methods, you relate peak area or, in some cases, peak height to concentration through calibration curves and internal standards.</p><h4><strong>Resolution</strong></h4><p>Resolution describes how well two peaks are separated and depends on efficiency, selectivity, and retention. Adequate resolution means peaks do not overlap significantly, which improves identification confidence and quantification for closely eluting impurities.</p><h4><strong>Dead Time (t₀)/Void Time</strong></h4><p>Dead time, <i>t</i><sub>0</sub>, or hold‑up time, <i>t</i><sub>M</sub> (or void time), is the time taken by unretained species or the mobile phase front to pass through the column and reach the detector. You use <i>t</i><sub>0</sub> as the reference for calculating capacity factor and for assessing how strongly analytes interact with the stationary phase.</p></div><div><div><div><div><h3><strong>Step‑by‑Step Guide to Reading an HPLC Chromatogram</strong></h3></div></div></div></div><div><p>Interpreting your data requires a systematic approach to guarantee you capture all critical details. Follow this sequential process to evaluate your chromatographic results confidently and accurately.</p><h4><strong>Identify the Baseline Stability</strong></h4><p>Start by checking if the baseline is flat or if you see random noise and slow drift across the run. Significant drift or high noise suggests issues with temperature control, solvent quality, or detector stability, which must be addressed before trusting quantitation.</p><h4><strong>Locate and Count Peaks</strong></h4><p>Next, scan the chromatogram to locate all distinct peaks above the baseline and note their relative sizes. Counting peaks versus expectations from standards or sample composition helps you detect co‑elution, extra peaks, or missing analytes.</p><h4><strong>Determine Retention Time</strong></h4><p>Measure retention time, <i>t</i><sub>R</sub>, for each peak and record these values alongside known standards or system-suitability data. Consistent <i>t</i><sub>R</sub> values within defined windows support correct identification, while larger shifts can indicate changes in temperature, mobile phase composition, or flow rate.</p><h4><strong>Calculate Capacity Factor (k') Using t<sub>M</sub> and t<sub>R</sub></strong></h4><p>Capacity factor, <i>K'</i> (or retention factor), describes how much longer an analyte is retained compared with unretained material and is calculated from the chromatogram. Using the dead time, <i>t</i><sub>M</sub>, (void peak) and analyte retention time, <i>t</i><sub>R</sub>, you apply<strong> </strong><i><strong>k</strong></i><strong> = (</strong><i><strong>t</strong></i><strong><sub>R</sub> – </strong><i><strong>t</strong></i><strong><sub>M</sub>)/</strong><i><strong>t</strong></i><strong><sub>M</sub></strong> to assess whether the analyte is sufficiently retained under the chosen conditions.</p><p>Knowing how to analyze HPLC results with capacity factor helps you judge if peaks elute too close to <i>t</i><sub>0</sub>, or spend an appropriate time interacting with the column, which guides method refinement.</p><h4><strong>Analyze Peak Area for Quantification (H3)</strong></h4><p>For quantification, integrate each peak to obtain the area and compare it with calibration standards or internal standards to calculate concentration. Reliable peak area measurements depend on good baseline definition and stable detector response, so always verify baseline quality before finalizing your measurements.</p><h4><strong>Evaluate Peak Shape and Resolution</strong></h4><p>Assess whether peaks are symmetric or show tailing, fronting, or broadening, and examine spacing between adjacent peaks for resolution. Changes in peak shape across the chromatogram can reveal extra‑column effects, packed bed issues, or interaction with active sites on the stationary phase.</p><p>If you see persistent problems, review your <a target="_blank" rel="noopener noreferrer" href="https://labrulez.com/en-us/lcms/products?category=48&amp;manufacturerItems=50"><strong><u>HPLC column</u></strong></a> choice, sample preparation, and gradient or isocratic settings to refine separation.</p></div><div><div><div><div><h3><strong>Common Problems in HPLC Chromatograms and How to Identify Them</strong></h3><p>Even well-developed methods can degrade over time and produce visual cues that signal system or chemistry faults. Recognizing these characteristic peak and baseline distortions is your first step in troubleshooting.</p><h4><strong>Peak Tailing</strong></h4><p><a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/resources/knowledge-center/hplc-knowledge-center/how-to-reduce-peak-tailing-in-hplc"><strong><u>Peak tailing</u></strong></a> appears when the right side of a peak stretches out into a long tail instead of dropping back to the baseline symmetrically. On the chromatogram, the trailing edge slopes slowly and the peak looks skewed to the left, which makes integration boundaries less obvious and can hint at dispersion or secondary interactions. You can reduce tailing by minimizing extra‑column volume and checking column packing and active sites.</p><h4><strong>Peak Fronting</strong></h4><p><a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/resources/knowledge-center/hplc-knowledge-center/peak-fronting"><strong><u>Peak fronting</u></strong></a> shows a steep leading edge and a compressed front portion of the peak, so the apex seems pulled forward relative to the rest of the peak profile. Visually, the peak looks like it is leaning in the direction of flow and the front overlaps more strongly with neighboring peaks, which can distort apparent purity for fast‑eluting components.</p><p>Consider lowering sample load or injection volume and matching diluent strength to the mobile phase, and when fronting persists, consult Phenomenex resources on <a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/resources/knowledge-center/gc-knowledge-center/peak-shape-problems-tailing-peaks"><strong><u>peak-shape problems</u></strong></a>.</p><h4><strong>Noise and Drift</strong></h4><p>Baseline noise appears as small, rapid fluctuations around the baseline that make it harder to see low‑level peaks. Baseline drift shows up as a slow rise or fall of the baseline across the run, so the line no longer stays flat; this can cause apparent changes in peak height and shifts in where you draw integration baselines. Simple checks like improving solvent degassing, stabilizing temperature, and cleaning detector flow cells help reduce noise and drift.</p><h4><strong>Co‑Elution</strong></h4><p>Co‑elution is easy to spot when two peaks do not fully separate and instead form a shoulder or merged profile on the chromatogram.You may see one peak with a plateau or a visible kink where two components overlap, and the valley between them does not return to the baseline, which complicates assignments for closely eluting impurities. To resolve co‑elution, adjust selectivity by changing mobile phase composition, gradient profile, or column chemistry.</p><h4><strong>Broad Peaks</strong></h4><p>Broad peaks have a noticeably wider base and lower height compared with well‑focused peaks of similar are. They often span more time units on the x‑axis and show gentle slopes on both sides, indicating band spreading that reduces resolution and makes small shoulders harder to detect. Narrowing broad peaks usually involves reducing extra‑column volumes and confirming column performance and particle size suitability.</p><h4><strong>Retention-Time Shifting</strong></h4><p>Retention-time shifting is seen when all peaks move earlier or later in the chromatogram compared with previous runs, while their shapes remain similar. When you overlay chromatograms, consistent shifts in peak positions along the time axis, often on the order of a few percent, indicate changes in conditions such as temperature or mobile phase that must be tracked carefully. Fixing retention time shifts starts with stabilizing temperature and verifying mobile phase composition and flow rate settings across runs.</p></div></div></div></div><div><div><div><div><h3><strong>Tips for Accurate HPLC Chromatogram Interpretation</strong></h3></div></div></div></div><div><p>To read HPLC chromatograms accurately, maintain a stable temperature, use well‑degassed mobile phases, and keep detector flow cells and optics clean. Regularly check capacity factor and selectivity values against method expectations, and verify that resolution is adequate for critical pairs before accepting results.</p><p>When retention times drift or peak shapes degrade, review your <a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/resources/knowledge-center/gc-knowledge-center/understanding-retention-time"><strong><u>retention-time</u></strong></a> trends alongside system maintenance logs so you can correct problems early.</p></div><div><div><div><div><h3><strong>FAQs on Reading HPLC Chromatograms</strong></h3></div></div></div></div><div><h4><strong>How Do You Calculate Capacity Factor (k') from a Chromatogram?</strong></h4><p>Capacity factor, <i>k</i>, is calculated using the analyte retention time, [Equation], and the dead time, <i>t</i><sub>R</sub>, measured from the unretained peak. Apply <i>k</i> = (<i>t</i><sub>R</sub> – <i>t</i><sub>M</sub>)/<i>t</i><sub>M</sub>, so higher k values indicate stronger interactions with the stationary phase and longer retention.</p><h4><strong>How Is Selectivity (α) Calculated Using Chromatogram Data?</strong></h4><p>Selectivity (α) describes how well two components are separated and is defined as the ratio of their retention factors, α = <i>k</i><sub>2</sub>/<i>k</i><sub>1</sub>, for peaks, where <i>k</i><sub>2</sub> &gt; <i>k</i><sub>1</sub>. Values higher than 1.2 show separation, and larger α values correspond to better selectivity between the peaks.</p><h4><strong>What Does Peak Area Indicate in HPLC?</strong></h4><p>Peak area reflects the total amount of analyte that reaches the detector and is generally more reliable than peak height for quantitative work. Build calibration curves that relate peak area to concentration, then apply them only when the baseline and detector response remain stable across all injections.</p><h4><strong>How Is Retention Time Used in HPLC Analysis?</strong></h4><p>Retention time is used as a qualitative index because each compound elutes at a characteristic time when conditions are held constant. Compare sample retention times with those of standards and then pair that information with peak area or height to confirm identity and calculate concentration in routine analyses.</p></div>

How to Read an HPLC Chromatogram: Step‑by‑Step Guide to Interpreting Results

<p><i><span><strong>Are you currently using single-column batch chromatography to isolate and purify target compounds, but struggling with high solvent demand, low product concentration, or limited throughput? Simulated Moving Bed chromatography offers an alternative approach that can address many of these limitations through continuous multi-column separation.</strong></span></i></p><h4><span><strong>What is Simulated Moving Bed Chromatography (SMB)?</strong></span></h4><p><span>Simulated Moving Bed (SMB) chromatography is a continuous multi-column separation technique designed to make more efficient use of the stationary phase while minimizing solvent consumption. It is particularly well suited to the purification of binary and pseudo-binary mixtures, including enantiomers, sugars, petrochemical products, and fine chemicals.</span></p><h4><span><strong>Conventional Batch Chromatography</strong></span></h4><p><span>In traditional batch chromatography, the sample is introduced onto a single chromatographic column and transported through the stationary phase by the mobile phase. Individual compounds interact with the stationary phase to different extents and therefore travel through the column at different rates:</span></p><ul><li><span>Weakly retained compounds migrate more rapidly.</span></li><li><span>Strongly retained compounds move through the column more slowly.</span></li></ul><p><span>The separated substances consequently leave the column at different retention times and can be collected as individual fractions (Fig. 1). Before a new sample is normally introduced, the previous sample must be completely eluted and the column prepared for the next separation.</span></p><p><span>Although single-column batch chromatography is versatile and comparatively easy to operate, several disadvantages become increasingly important at production scale. The stationary phase is not continuously involved in useful separation, which can limit productivity. Other drawbacks include substantial solvent consumption, diluted product fractions, repeated column regeneration, and potential loss of valuable product.</span></p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_1_Single_column_separation_A_fast_eluting_component_B_retaining_component_cda8ef66ef.jpg" alt="KNAUER: Figure 1: Single column separation; A – fast eluting component, B – retaining component" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_1_Single_column_separation_A_fast_eluting_component_B_retaining_component_cda8ef66ef.jpg 76w," sizes="100vw" width="394" height="800"></p><h4><span><strong>From True Moving Bed to SMB</strong></span></h4><p><span>The principle behind SMB originates from the theoretical <strong>True Moving Bed (TMB)</strong> process. In an ideal TMB system, the liquid mobile phase and the solid stationary phase continuously travel in opposite directions. This counter-current arrangement takes advantage of differences in the affinity of individual compounds toward the two phases.</span></p><p><span>Consider a binary mixture containing components A and B. Component A interacts more strongly with the stationary phase, whereas component B is less strongly retained and preferentially follows the liquid mobile phase. Under appropriate operating conditions, the two substances effectively migrate in opposite directions and can be collected continuously through separate outlets (Fig. 2).</span></p><p><span>While this concept is highly efficient from a theoretical perspective, continuously moving a packed solid chromatographic adsorbent is technically difficult to realize.</span></p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_2_True_Moving_Bed_concept_e1eac66028.jpg" alt="KNAUER: Figure 2: True Moving Bed concept" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_2_True_Moving_Bed_concept_e1eac66028.jpg 186w," sizes="100vw" width="954" height="800"></p><p><span>Four process ports divide the system into four functional zones (Zone I–IV):</span></p><ol><li><span>Eluent (desorbent) inlet</span></li><li><span>Extract outlet</span></li><li><span>Feed inlet</span></li><li><span>Raffinate outlet</span></li></ol><p><span>Feed and eluent are introduced continuously, while two product streams are simultaneously withdrawn:</span></p><ul><li><span><strong>Extract</strong> – the strongly retained, later-eluting component</span></li><li><span><strong>Raffinate</strong> – the less strongly retained, earlier-eluting component</span></li></ul><h4><span><strong>The Four Functional Zones</strong></span></h4><p><span>Each of the four zones has a defined purpose and operates at its own internal flow rate (VI–VIV):</span></p><ul><li><span><strong>Zone I</strong> regenerates the stationary phase.</span></li><li><span><strong>Zone II</strong> prevents the weakly retained component from entering the extract stream.</span></li><li><span><strong>Zone III</strong> prevents the strongly retained component from contaminating the raffinate.</span></li><li><span><strong>Zone IV</strong> enables recovery and internal reuse of the remaining solvent.</span></li></ul><h4><span><strong>How is the TMB Principle Implemented in SMB?</strong></span></h4><p><a target="_blank" rel="noopener noreferrer" href="https://www.knauer.net/smb"><span>SMB</span></a><span> eliminates the need to physically move the stationary phase. Instead, the adsorbent remains fixed inside the chromatographic columns, while the positions of the liquid inlet and outlet ports are periodically shifted.</span></p><p><span>From the perspective of these moving connection points, the stationary phase appears to travel in the opposite direction to the liquid phase. SMB therefore reproduces the counter-current behavior of a TMB system without physically transporting the packed adsorbent (Fig. 3).</span></p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_3_Transfer_TMB_concept_to_SMB_concept_e4b8905269.jpg" alt="KNAUER: Figure 3: Transfer TMB concept to SMB concept" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_3_Transfer_TMB_concept_to_SMB_concept_e4b8905269.jpg 245w," sizes="100vw" width="1372" height="800"></p><p><span>A typical SMB setup consists of several chromatographic columns arranged in a closed loop. A standard configuration often uses eight columns, although the number can be adapted according to the application.</span></p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_4_SMB_concept_fb005cdfa1.jpg" alt="KNAUER: Figure 4: SMB concept" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_4_SMB_concept_fb005cdfa1.jpg 195w," sizes="100vw" width="1001" height="800"></p><p><span>After a predefined switching time, all inlet and outlet positions move forward by one column in the direction of the mobile-phase flow. The columns themselves stay in place, but their functional roles within the process change.</span></p><p><span>By repeatedly shifting the ports, the system creates the effect of a stationary phase moving counter-currently to the mobile phase. With eight columns, eight switching steps are required to complete one full SMB cycle.</span></p><p><span>Following an initial transition period, the process reaches a cyclic steady state. Concentrations continue to fluctuate during individual switching intervals, but the overall concentration profile repeats after every complete cycle.</span></p><p><span>Increasing the number of columns and reducing the size of individual switching steps allows the SMB system to reproduce ideal TMB behavior more closely. Systems containing fewer columns exhibit more pronounced discrete switching effects and can require more sophisticated dynamic modeling during process development.</span></p><h3><span><strong>What are the Main Advantages of SMB?</strong></span></h3><h4><span><strong>1. Continuous Operation and Higher Productivity</strong></span></h4><p><span>The key benefit of SMB chromatography is continuous operation. Feed and eluent enter the system continuously, while purified extract and raffinate streams are removed at the same time.</span></p><p><span>As a result, SMB systems can operate for prolonged periods—from several days to even several weeks—with very limited interruption.</span></p><p><span>Because several chromatographic columns perform different separation functions simultaneously, the stationary phase is utilized much more effectively than in conventional batch chromatography. This can translate into higher productivity, improved process efficiency, and a continuous supply of purified product.</span></p><h4><span><strong>2. Reduced Solvent Consumption</strong></span></h4><p><span>Conventional batch chromatography can require substantial volumes of mobile phase to achieve separation and product recovery. The collected fractions are often highly diluted, creating an additional need for downstream concentration.</span></p><p><span>SMB maintains concentration profiles within the circulating system and uses the mobile phase more efficiently. Partial internal recycling of solvent is an inherent part of the process.</span></p><p><span>Lower solvent consumption can lead to:</span></p><ul><li><span>Reduced raw-material costs</span></li><li><span>Lower volumes of liquid waste</span></li><li><span>Reduced energy requirements for solvent evaporation</span></li><li><span>More concentrated product streams and less downstream concentration</span></li></ul><h4><span><strong>3. High Recovery and Reproducible Product Quality</strong></span></h4><p><span>Once the process reaches cyclic steady state, SMB can continuously generate product streams with stable and reproducible composition.</span></p><p><span>Additional advantages can include high product recovery, lower product dilution, more efficient use of the stationary phase, and straightforward integration into continuous manufacturing strategies.</span></p><h4><span><strong>What are the Differences Between Batch LC and SMB?</strong></span></h4><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_differences_between_batch_LC_and_SMB_eba3897f65.jpg" alt="KNAUER: Differences between batch LC and SMB" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_differences_between_batch_LC_and_SMB_eba3897f65.jpg 245w," sizes="100vw" width="1061" height="638"></p><h4><span><strong>Where is SMB Used?</strong></span></h4><p><span>SMB technology is established across several industries and can be applied to purification processes ranging from gram-scale production to several tons per day (Fig. 5).</span></p><p><span>Typical and emerging applications include:</span></p><ul><li><span>Sugar purification</span></li><li><span>Petrochemical separations</span></li><li><span>Chiral separations</span></li><li><span>Fine chemical production</span></li><li><span>Pharmaceutical intermediates</span></li><li><span>Selected biomolecule separations</span></li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Figure_5_SMB_application_areas_7109ff703f.jpg" alt="KNAUER: Figure 5: SMB application areas; author's illustration" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Figure_5_SMB_application_areas_7109ff703f.jpg 245w," sizes="100vw" width="1500" height="512"></p><h4><span><strong>Limitations of SMB</strong></span></h4><p><span>Although SMB can provide substantial advantages, it is not necessarily the ideal solution for every chromatographic separation.</span></p><p><span>The instrumentation and process configuration are more complex than conventional single-column chromatography. Several pumps, valves, columns, and liquid pathways must be controlled precisely. Successful method development therefore requires a solid understanding of adsorption equilibria, mass-transfer effects, and fluid dynamics.</span></p><p><span>Classical four-zone SMB is primarily intended for binary and pseudo-binary separations. More complicated mixtures may require modified SMB configurations, additional purification steps, or multiple coupled SMB systems.</span></p><p><span>SMB is generally most economically attractive when relatively large quantities of a consistent feed material are processed continuously over extended periods.</span></p><p><span>Batch chromatography can remain the better option for small-scale production, frequent changes between products, complex multicomponent mixtures, or applications that depend on flexible gradient separations.</span></p><h4><span><strong>Conclusion</strong></span></h4><p><span>Simulated Moving Bed chromatography converts the theoretical counter-current principle of True Moving Bed separation into a practical continuous chromatographic process. Instead of physically transporting the stationary phase, SMB periodically changes the positions of the feed, eluent, extract, and raffinate connections. This reproduces the effect of counter-current movement between stationary and mobile phases.</span></p><p><span>Compared with conventional batch chromatography, SMB can provide higher productivity, more efficient stationary-phase utilization, substantially lower solvent consumption, and more concentrated product streams. Continuous operation makes the technology particularly attractive for demanding large-scale binary separations, including the purification of pharmaceutical enantiomers.</span></p><p><span>Although successful SMB implementation requires accurate process data, careful method development, and precise control of operating conditions, the technology demonstrates how an idealized chromatographic principle can be transformed into a robust and efficient industrial separation strategy.</span></p>

Simulated Moving Bed Chromatography: Efficient Continuous Separation

News from LabRulezLCMS Library - Week 44, 2026

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Phenomenex is a global technology leader committed to developing novel analytical chemistry solutions that solve the separation and purification challenges of researchers in industrial, government and academic laboratories.

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

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Waters SELECT SERIES Cyclic IMS

The SELECT SERIES Cyclic IMS combines novel cyclic ion mobility separation with state-of-the-art, high performance, time-of-flight mass spectrometry.

<div><div id="title-c6692d35e8" data-locator="title-with-text"><h3><strong>Endless possibilities. Limitless research.</strong></h3></div></div><div><div id="text-ba9d3d94da" data-locator="text"><p><strong>Developed through focused collaboration, the SELECT SERIES Cyclic IMS combines novel cyclic ion mobility separation with state-of-the-art, high performance, time-of-flight mass spectrometry, enabling leading researchers to unlock the potential in scientific discovery.</strong></p><p>The SELECT SERIES Cyclic IMS enables selection of ions by both mass and mobility, a unique functionality of <strong>Cyclic Ion Mobility (cIM)</strong> that provides unparalleled flexibility and capabilities for research studies in both academia and industry. With ion mobility and mass spectrometry, you can separate by shape as well as&nbsp;<i>m/z</i>&nbsp;for separation of isomers, measure the <strong>Collisional Cross Section (CCS)</strong> of an ion, and achieve significantly cleaner mass spectral data.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_SELECT_SERIES_Cyclic_IMS_Novel_Cyclic_Ion_Mobility_c_IM_Technology_13abf6dd9b.jpg" alt="Waters SELECT SERIES Cyclic IMS: Novel Cyclic Ion Mobility (cIM) Technology." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_SELECT_SERIES_Cyclic_IMS_Novel_Cyclic_Ion_Mobility_c_IM_Technology_13abf6dd9b.jpg 234w," sizes="100vw" width="1024" height="682"></p><h3><strong>Overview</strong></h3><div data-locator="section-container-body"><div><div><div data-locator="layoutcontainer-"><div><div id="text-26eab4b89b" data-locator="text"><ul><li>Combine <strong>cyclic ion mobility (cIM) separation</strong> with <strong>ToF mass spectrometry</strong></li><li>Separate more than ever before with ultimate ion mobility resolution for complete confidence</li><li>Perform <strong>unique IMS<sup>n</sup> experiments</strong> for increased selectivity and advanced characterization</li><li>Enable <strong>pre-mobility mass selection</strong> and <strong>mobility selection of precursor and product ions</strong></li><li>Achieve increased experimental flexibility for solving complex problems with three distinct fragmentation regions</li></ul></div></div><div><div id="text-36ff553b5f" data-locator="text"><h3><strong>Recommended Use</strong></h3><p>For identifying and rapidly monitoring co-eluting, isomeric metabolites in chemical reactions</p><blockquote><p class="cke-user_style-important"><strong>Video: </strong><a target="_blank" rel="noopener noreferrer" href="https://www.youtube.com/watch?v=nauwVH0L91o"><strong>Waters SELECT SERIES Cyclic IMS</strong></a></p></blockquote><div><div id="text-8469d7776e" data-locator="text"><h3 data-locator="modal-title"><strong>Discover more with ultimate ion mobility resolution</strong></h3><p>The instrument design and control software enable a range of experiments, including acquisition of Tof data, Single pass ion mobility, and the ability to scale mobility resolution to match the complexity of a sample. Ion mobility resolution increases with the length of the mobility device and therefore with the number of passes.</p><figure class="image"><img style="aspect-ratio:1024/682;" src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_SELECT_SERIES_Cyclic_IMS_IMS_resolution_increases_with_the_number_of_passes_around_the_c_IM_device_121719dbe0.jpg" alt="Waters SELECT SERIES Cyclic IMS: IMS resolution increases with the number of passes around the cIM device. Demonstrated here by the separation of reverse sequence peptides SDGRG and GRGDS. The Arrival Time Distributions (ATDs) obtained after 100 passes around the device are inset." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_SELECT_SERIES_Cyclic_IMS_IMS_resolution_increases_with_the_number_of_passes_around_the_c_IM_device_121719dbe0.jpg 234w," sizes="100vw" width="1024" height="682"></figure><div><div id="text-bb2fb9f91c" data-locator="text"><h3 data-locator="modal-title"><strong>Unlock the potential in scientific discovery</strong></h3><p>The unique instrument design of the SELECT SERIES Cyclic IMS&nbsp;facilitates advanced ion mobility experiments like IMS<sup>n</sup>&nbsp;isolation. IMS<sup>n</sup>&nbsp;isolation&nbsp;extends the resolution of a specific mobility range,&nbsp;<i>e.g.,</i>&nbsp;ions of a specific species. Eject ions (without acquisition) outside of the desired mobility range, leaving the remaining species in the cIM to continue the separation process. This “trimming” approach can be undertaken numerous times and can be used, for example, to increase specificity of the selected ions or to prevent “wrap-around” in the cIM where high mobility ions catch up with low mobility ions in multi-pass experiments. Multiple cyclic sequences of different numbers of passes can be defined in one, multi-pass LC-MS method, increasing productivity by reducing the need for multiple injections of the same sample.</p></div></div><figure class="image"><img style="aspect-ratio:1024/578;" src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_SELECT_SERIES_Cyclic_IMS_Investigation_into_acyl_glucuronides_via_infusion_f3bf2c67e2.jpg" alt="Waters SELECT SERIES Cyclic IMS: Investigation into acyl glucuronides via infusion. Slicing enables further IMS separation of target metabolite (blue region) of interest and other ions can be ejected. Increasing the IMS resolution reveals an additional 4-O acyl. In collaboration with Imperial College London – Ian D. Wilson." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_SELECT_SERIES_Cyclic_IMS_Investigation_into_acyl_glucuronides_via_infusion_f3bf2c67e2.jpg 245w," sizes="100vw" width="1024" height="578"></figure><div><div id="text-78a4dc4d5e" data-locator="text"><h3 data-locator="modal-title"><strong>Undertake advanced structural characterization</strong></h3><p><strong>IMS<sup>n</sup>&nbsp;activation</strong>&nbsp;is ideal for <strong>advanced structural characterization</strong>. During mobility separation, excise a segment of the mobility separated species and send the ions to the pre array storage device. Unwanted ions are ejected, and the stored ions are then re-injected into the array under collision induced activation/dis-sociation conditions, as required, and subsequent mobility separation performed. The product ions can then be sent to the&nbsp;Tof<i>&nbsp;</i>for detection or can undergo further rounds of mobility selection providing the unique IMS<sup>n</sup>&nbsp;capability.</p></div></div><div><div id="text-079d02ceb8" data-locator="text"><h3 data-locator="modal-title"><strong>Imaging mass spectrometry with DESI XS and Cyclic IMS</strong></h3><p>The unique combination of&nbsp;<a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?instrumentationItems=26&amp;mainauthorItems=5&amp;search=DESI+XS"><strong>DESI XS</strong></a><strong>&nbsp;</strong>with SELECT SERIES Cyclic IMS provides ultimate levels of specificity for <strong>MS imaging</strong> approaches. DESI XS produces a visual map of the spatial distribution of a wide range of analytes, for example small molecule drugs, metabolites, and lipids across the surface of a tissue sample, and the unique capabilities and modes of acquisition of Cyclic IMS enable comprehensive in-depth experiments to be performed.</p></div></div><div><div id="text-673d117887" data-locator="text"><h3 data-locator="modal-title"><strong>Increased sensitivity provides confidence in what you find</strong></h3><p>To complement the novel <strong>Cyclic Ion Mobility</strong> capability and for complete confidence in your data, the&nbsp;SELECT SERIES&nbsp;Cyclic IMS is equipped with the latest <strong>ToF technology</strong>. The XS transfer device maintains the fidelity of the ion mobility separation while simultaneously conditioning the ion beam for the absolute best time-of-flight performance. The new offset oa-Tof with dual gain detection system <strong>provides improved sensitivity, resolution &gt;100,000 (FWHM), reliable accurate exact mass measurements</strong> (low ppm), and <strong>improved dynamic range</strong>.</p><figure class="image"><img style="aspect-ratio:1024/682;" src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Mass_spectrum_highlighting_resolution_of_100_000_FWHM_on_the_7_charge_state_of_monomeric_9ff8f53204.jpg" alt="Waters SELECT SERIES Cyclic IMS: Mass spectrum highlighting resolution of >100,000 (FWHM) on the 7+ charge state of monomeric." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Mass_spectrum_highlighting_resolution_of_100_000_FWHM_on_the_7_charge_state_of_monomeric_9ff8f53204.jpg 234w," sizes="100vw" width="1024" height="682"></figure></div></div></div></div></div></div></div></div></div></div></div></div>

Waters Cyclic IMS P20 Mass Spectrometer

Cyclic IMS P20 combines high-resolution ion mobility, native MS, and imaging capabilities for detailed structural characterization of complex biological samples.

<p><strong>The Waters Cyclic IMS P20 Mass Spectrometer is a next-generation ion mobility-enabled mass spectrometry platform designed to reveal molecular structure with greater confidence by combining high-resolution cyclic ion mobility separation with accurate mass analysis. The system resolves co-eluting, isomeric, and isobaric species, providing deeper structural insight into complex biological samples.</strong></p><p>Built for advanced research applications, the Cyclic IMS P20 supports proteomics, metabolomics, lipidomics, native mass spectrometry, and mass spectrometry imaging workflows. Multi-pass ion mobility separations, Wideband Enhancement (WBE), and advanced fragmentation technologies provide exceptional analytical depth and sensitivity.</p><p>The platform supports <strong>MALDI XS and DESI XS imaging</strong> workflows, enabling high-confidence spatial analysis and molecular visualization. Additional capabilities including CIU, HDX, ECD, and SID make the system ideally suited for investigating biomolecular structure, stability, dynamics, and intermolecular interactions.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_Cyclic_IMS_P20_Mass_Spectrometer_with_MALDI_XS_and_DESI_XS_1a6ac479a1.jpg" alt="Waters: Cyclic IMS P20 Mass Spectrometer with MALDI XS and DESI XS" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_Cyclic_IMS_P20_Mass_Spectrometer_with_MALDI_XS_and_DESI_XS_1a6ac479a1.jpg 234w," sizes="100vw" width="1024" height="682"></p><h3><strong>Key Features</strong></h3><ul><li>High-resolution Cyclic Ion Mobility Spectrometry</li><li>Separation of isomeric, isobaric, and co-eluting species</li><li>Multi-pass ion mobility separation</li><li>Wideband Enhancement (WBE) technology</li><li>Up to 10× improvement in MS/MS sensitivity</li><li>Advanced native MS capabilities</li><li>Dynamic Field Declustering (DFD)</li><li>Quadrupole mass selection up to 32,000 m/z</li><li>Extended mass range beyond 100,000 m/z</li><li>Electron Capture Dissociation (ECD)</li><li>Surface-Induced Dissociation (SID)</li><li>Integrated Collision-Induced Unfolding (CIU)</li><li>Hydrogen-Deuterium Exchange (HDX) workflows</li><li>MALDI XS and DESI XS imaging support</li><li>Automated CCS calibration</li><li>Intuitive software environment</li><li>Modular and future-proof platform</li></ul><h3><strong>Technical Specifications</strong></h3><h4><strong>Technology</strong></h4><ul><li>Cyclic Ion Mobility Spectrometry (Cyclic IMS)</li><li>Orthogonal Acceleration Time-of-Flight (oaTOF)</li><li>Multi-pass ion mobility separation</li><li>IMSⁿ experiments</li></ul><h4><strong>Advanced Capabilities</strong></h4><ul><li>Wideband Enhancement (WBE)</li><li>Dynamic Field Declustering (DFD)</li><li>Collision-Induced Unfolding (CIU)</li><li>Electron Capture Dissociation (ECD)</li><li>Surface-Induced Dissociation (SID)</li><li>Hydrogen-Deuterium Exchange (HDX)</li></ul><h4><strong>Mass Range</strong></h4><ul><li>Quadrupole selection up to 32,000 m/z</li><li>Detection beyond 100,000 m/z</li></ul><h4><strong>Imaging Applications</strong></h4><ul><li>MALDI XS</li><li>DESI XS</li><li>High-resolution molecular imaging</li><li>Isomeric lipid separation</li></ul><h4><strong>Software</strong></h4><ul><li>Automated CCS calibration</li><li>Advanced method development</li><li>IMS data processing and interpretation</li><li>HDX and CIU workflow support</li></ul><h4><strong>Typical Applications</strong></h4><ul><li>Proteomics</li><li>Native mass spectrometry</li><li>Structural biology</li><li>Lipidomics</li><li>Metabolomics</li><li>Biotherapeutic characterization</li><li>GLP-1 research</li><li>Protein complex analysis</li><li>Molecular imaging</li><li>Forensics</li><li>Environmental analysis</li></ul>

Waters SYNAPT XS High Resolution Mass Spectrometer

The SYNAPT XS Ion Mobility Time-of-Flight Mass Spectrometer provides ultimate flexibility, offering greater freedom of analytical choice to support scientific creativity and technical success for any application.

<div><div><h4><strong>Ultimate flexibility for deeper discovery</strong></h4></div></div><div><div><p><strong>In the arena of scientific discovery, the slow pace of advancement and escalating costs of research can prove to be a challenge. Unlike competitor systems with restricted inlet options, scan function limitations, or requiring multiple platforms, only Waters offers an all-encompassing high-performing LC-MS solution, the SYNAPT XS mass spectrometer, for ultimate flexibility and greater freedom of analytical choice – supporting scientific creativity and technical success for any application.</strong></p><ul><li><strong>Novel technology building blocks deliver cutting-edge analytical performance</strong></li><li><strong>SONAR</strong> and <strong>HDMSE</strong> provide a unique toolkit for interrogating complex mixtures</li><li><strong>Ion mobility</strong> multiplies peak capacity and selectivity of analysis</li><li><strong>CCS measurements</strong> increase security of compound identification</li><li><strong>Deeper discovery in Mass Spectrometry Imaging</strong> with<strong> </strong><a target="_blank" rel="noopener noreferrer" href="https://www.waters.com/waters/Full-Spectrum-Molecular-Imaging-/nav.htm?cid=135070661"><strong>Full Spectrum Molecular Imaging</strong></a> enabled by <a target="_blank" rel="noopener noreferrer" href="https://www.waters.com/waters/DESI-XS---Desorption-Electrospray-Ionization-/nav.htm?cid=135047466"><strong>DESI XS</strong></a> and MALDI</li></ul><h4><strong>Overview</strong></h4><div><div><div><div><ul><li><strong>Serve the broadest range of applications </strong>with the most extensive range of chromatograph inlets for ultimate flexibility</li><li><strong>Attain fragment ion information for every detectable component</strong> for unambiguous confirmation of compound identity with DIA modes <strong>SONAR</strong>, MSE, and <strong>HDMSE</strong></li><li><strong>Generate rich fragmentation data</strong> from highly complex samples in all acquisition modes including DDA</li><li>Simplify and streamline the mass spectral imaging workflow and deliver multi-layered, information-rich data with multiple imaging techniques and accompanying High Definition Imaging (HDI) Software</li><li><strong>Achieve automated higher order structure determination </strong>thorough confident identification, intact mass analysis, and best-in-class reproducibility</li><li><strong>Enhance MS/MS possibilities</strong> using complementary fragmentation techniques with high resolution and accurate mass measurements</li></ul></div></div><div><div><h5><strong>Recommended Use</strong></h5><p>For deeper discovery by aligning complimentary workflows on a single <strong>Ion mobility</strong> enabled time-of-flight mass spectrometer with the flexibility of multiple ion sources and acquisition modes.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_SYNAPT_XS_High_Resolution_Mass_Spectrometer_4_696d77ce2b.jpg" alt="Waters SYNAPT XS High Resolution Mass Spectrometer: The Waters SYNAPT XS offers flexibility and freedom of analytical choice with an extensive range of chromatographic inlets." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_SYNAPT_XS_High_Resolution_Mass_Spectrometer_4_696d77ce2b.jpg 234w," sizes="100vw" width="1024" height="682"></p><div><div><h3><strong>Cutting edge performance with novel technology</strong></h3><p>With technology building blocks inherited from the <strong>Waters Advanced Mass Spectrometry</strong> <strong>SELECT SERIES</strong>, advanced innovation is built-in, ensuring scientists using this platform are at the cutting edge of mass spectrometry while retaining the ease of use and established end-to-end workflows for which the <strong>SYNAPT </strong>is well known.</p><ul><li><strong>StepWave&nbsp;XS -&nbsp;</strong>Redesigned segmented quadrupole transfer optics provide enhanced sensitivity for challenging compounds while further improving levels of analytical robustness.</li><li><strong>Extended ToF -&nbsp;</strong>Delivers UPLC-compatible mass resolution, matrix-tolerant dynamic range, and quantitative results for the most complex samples, while offering superior specifications to previous SYNAPT models.</li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_SYNAPT_XS_High_Resolution_Mass_Spectrometer_9f19abb382.jpg" alt="Waters SYNAPT XS High Resolution Mass Spectrometer: SYNAPT XS schematic." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_SYNAPT_XS_High_Resolution_Mass_Spectrometer_9f19abb382.jpg 245w," sizes="100vw" width="1024" height="546"></p></div></div><div><div><h3><strong>Interrogate complex mixtures with a unique toolkit</strong></h3><p>A complete analytical strategy requires the combination of appropriate complementary techniques to see the bigger picture. With both <strong>SONAR</strong> and IMS-based <strong>data independent acquisition</strong> (DIA) modes of operation on the SYNAPT XS, the discovery analyst is in a unique position to interrogate complex mixtures by complementary mechanisms. Both types of acquisition increase analytical <strong>peak capacity</strong>, providing ‘clean and clear’ fragmentation data, but based on different molecular properties. This provides a truly unique investigative toolbox for the in-depth interrogation of complex mixtures.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_SYNAPT_XS_High_Resolution_Mass_Spectrometer_2_9179e575a4.jpg" alt="Waters SYNAPT XS High Resolution Mass Spectrometer: During each scan the quadrupole separates co-eluting precursor masses by scanning the mass range, and transiting them in sequence. Fragment ions from different precursors can then be recorded separately and assigned with confidence." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_SYNAPT_XS_High_Resolution_Mass_Spectrometer_2_9179e575a4.jpg 245w," sizes="100vw" width="1024" height="593"></p></div></div><div><div><h3><strong>Discover more with ion mobility and CSS measurement</strong></h3><p>Conventional mass spectrometers separate components based on m/z. The SYNAPT XS also allows separation of molecules according to<strong> their size, shape, and charge during ion mobility experiments as a function of their collisional cross section</strong> (CCS). As well as ion mobility offering an additional dimension of resolution, multiplying <strong>peak capacity</strong>, and <strong>selectivity of analysis</strong> the measurement of CCS offers additional molecular identifiers. The measured CCS of an ion can be used to help confirm an ion’s identity or investigate its structure. The inclusion of ion mobility significantly increases the extent and confidence with which a scientist can profile complex mixtures and complex molecules – dramatically enhancing sample definition.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_SYNAPT_XS_High_Resolution_Mass_Spectrometer_3_2240bff527.jpg" alt="Waters SYNAPT XS High Resolution Mass Spectrometer: Retention and drift time principle ion mobility enabled data-independent LC-HDMSE analysis (LC-DIA-IM-MS)." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_SYNAPT_XS_High_Resolution_Mass_Spectrometer_3_2240bff527.jpg 204w," sizes="100vw" width="1024" height="784"></p></div></div><div><div><h3><strong>Achieve ultra-confident structural characterization of components</strong></h3><p>Time aligned parallel (TAP) fragmentation is an acquisition mode unique to the T-Wave IMS design. It permits users to take the advantage of the <strong>TriWave configuration</strong>, which allows the pre-IMS T-Wave and post-IMS T-Wave to operate as two separate collision cells. The resultant CID-IMS-CID instrumental operation helps to provide ultra-confident structural characterization of components. <strong>TAP fragmentation</strong> provides a distinct advantage for building a complete structure, through superior fragment ion coverage, sensitivity, and accuracy compared to traditional MSⁿ or MS/MS techniques.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_SYNAPT_XS_High_Resolution_Mass_Spectrometer_5_4dd33f1ab5.jpg" alt="Waters SYNAPT XS High Resolution Mass Spectrometer: Schematic showing Time Aligned Parallel Fragmentation. " srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_SYNAPT_XS_High_Resolution_Mass_Spectrometer_5_4dd33f1ab5.jpg 234w," sizes="100vw" width="1024" height="682"></p></div></div></div></div></div></div></div></div>

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