<div><p><strong>Air bubbles in an HPLC system can disrupt pumping reliability, cause baseline noise, and result in poor result reproducibility and valve function. To prevent these, proper mobile phase degassing is essential.&nbsp;</strong></p><p>Degassing in HPLC not only ensures stable pumping but also improves sensitivity and accuracy by preventing dissolved oxygen from forming bubbles that interfere with detectors such as UV-VIS, fluorescence, refractive index, or electrochemical. Effective HPLC solvent degassing is therefore a critical step for stable, accurate, and reproducible chromatography.</p></div><div><div><div><div><h3><strong>What is HPLC Solvent Degassing and Why is it Necessary?</strong></h3></div></div></div></div><div><p>When solvents such as water, buffers, acetonitrile, and methanol contact the atmosphere, nitrogen and oxygen dissolve into them. When mixed, their combined air content can exceed the solubility of the mixture, creating a supersaturated condition that leads to outgassing and bubble formation. These bubbles disrupt <a target="_blank" rel="noopener noreferrer" href="https://www.phenomenex.com/techniques/hplc">HPLC</a> performance by causing unstable baselines, bad peak shape, pump cavitation, retention time shifting, and loss of sensitivity.</p><p>In pumps, air can stop flowing or cause erratic retention times, while in optical detectors, bubbles scatter light and generate noise or false signals. Although it is not necessary to remove all dissolved air, degassing in HPLC lowers gas levels below supersaturation and prevents these issues. Low-pressure mixing systems are especially prone to bubble formation, whereas high-pressure mixing reduces outgassing because solvents blend under elevated pressure.</p></div><div><div><div><div><h3><strong>Types of Degassing Techniques in HPLC</strong></h3></div></div></div></div><div><p>Degassing the mobile phase was once a major concern in liquid chromatography with outgassing problems being among the most common issues during the first decades of its use. The introduction of inline degassers in HPLC in the past 15–20 years has significantly reduced the occurrence of these problems, to the point that many users today have little experience with them. However, while inline degassing prevents most solvent outgassing issues, it does not eliminate all problems caused by dissolved air.</p></div><div><div><div><div><h4><strong>Inline Degassing</strong></h4></div></div></div></div><div><p>Inline degassing works by passing the mobile phase through a gas-permeable membrane within a vacuum chamber, allowing dissolved gases to escape while retaining the liquid. Different techniques include:</p><ul><li><strong>Vacuum degasser:</strong> Inline vacuum degassers, now standard in most modern HPLC systems, have greatly reduced bubble-related pump issues by efficiently removing dissolved gases. Solvents pass through porous tubing within a vacuum chamber, where gases diffuse out through micropores to waste, while the degassed liquid continues flowing onto the pump.</li><li><strong>Membrane degasser:</strong> Over the years, various polymer materials have been tried with varying success for degassers. Modern degassers use a thin, highly gas-permeable membrane, making the degassing process very efficient.</li></ul></div><div><div><div><div><h5><strong>Advantages</strong></h5></div></div></div></div><div><p>Inline degassers are highly reliable and effective at reducing dissolved gases in the mobile phase to acceptable levels and are now standard in most HPLC systems. It is usually automatic and trouble-free. They require little to no maintenance, making them convenient and reliable for routine use.</p></div><div><div><div><div><h5><strong>Limitations</strong></h5></div></div></div></div><div><p>Despite their effectiveness, inline degassers do not remove all dissolved air from the mobile phase. The residual air can still cause problems in certain sensitive applications. Membrane replacement, vacuum pump failure, or damaged tubing can also cause issues, so proper maintenance is essential to avoid costly repairs. Additionally, microbial growth in the buffer can block membrane pores and contaminate the entire HPLC system. To prevent microbial growth, contamination, or blockages, it is best not to store the degasser with water or buffer in the tubing and to flush with 30% phosphoric acid followed by water if contamination occurs.</p></div><div><div><div><div><h4><strong>Offline Degassing</strong></h4></div></div></div></div><div><p>Offline degassing involves removing dissolved air from the mobile phase before it enters the HPLC system. The techniques include:</p><ul><li><strong>Helium sparging:</strong> This method requires a solvent inlet filter to prevent particulates and improve gas dispersion, but the degassed solvent can reabsorb air once in the system. To reduce bubble formation, slight back pressure should be maintained, often using a backpressure regulator after the detector. Helium purging can remove <strong>up to 80% of dissolved air</strong>, requiring equal helium volume for organic–aqueous phases. However, the flow rate should be reduced over time to avoid losing volatile components.</li><li><strong>Vacuum filtration:</strong> Vacuum degassing removes <strong>over 60%</strong> of dissolved air from the mobile phase and is performed through batch-wise treatment. It can also be done during filtration, before the chromatographic process with a 0.45 or 0.22 μm membrane filter, though this method is less effective.</li><li><strong>Sonication:</strong> Ultrasonic baths alone are not effective for degassing, as they remove<strong> only about 20-30% of dissolved gases</strong>, while at least 50% removal is needed. However, when combined with helium sparging or vacuum degassing, sonication becomes a <strong>useful complementary method</strong>. Since ultrasonic baths are common in laboratories, they are best applied alongside other degassing techniques.</li></ul></div><div><div><div><div><h5><strong>Advantages</strong></h5></div></div></div></div><div><p>An offline degasser offers the advantage of using a 2 µm solvent inlet filter to prevent particulates from entering the mobile phase while ensuring efficient gas dispersion. It is also cost-effective and flexible, as it relies on standard laboratory equipment and can handle large solvent volumes in a batch.</p></div><div><div><div><div><h5><strong>Limitations</strong></h5></div></div></div></div><div><p>A major drawback of offline helium sparging is that the mobile phase can gradually reabsorb air once placed in the HPLC system, leading to signal loss and changing sensitivity. Additionally, helium sparging can be expensive as it requires specialized gas cylinders.</p><h3><strong>Inline vs Offline Degassing: Side-by-Side Comparison</strong></h3><h5><strong>Timing:</strong></h5><ul><li><strong>Offline Degassing: </strong>It is usually done before the mobile phase enters the HPLC run.</li><li><strong>Inline Degassing: </strong>It is a continuous process that occurs during an HPLC run.</li></ul><h5><strong>Degassing Efficiency:</strong></h5><ul><li><strong>Offline Degassing:&nbsp;</strong>The efficiency can be variable. For instance, vacuum/sonication removes 60–70% of dissolved air, sonication alone removes <strong>20–30%</strong> of air and helium sparging removes 80% of air.</li><li><strong>Inline Degassing:&nbsp;</strong>It does not remove 100% of dissolved gas, but it removes most of it. However, it prevents bubble formation.</li></ul><h5><strong>Risk of Re-dissolving:</strong></h5><ul><li><strong>Offline Degassing:&nbsp;</strong>The risk is high as solvents can reabsorb air when exposed to the atmosphere.</li><li><strong>Inline Degassing:&nbsp;</strong>The risk is low as the continuous vacuum keeps new gas from dissolving.</li></ul><h5><strong>Cost:</strong></h5><ul><li><strong>Offline Degassing:&nbsp;</strong>Low initial investment and requires only basic lab equipment. <strong>Helium</strong> sparging can be expensive as it requires specialized gas cylinders.</li><li><strong>Inline Degassing:&nbsp;</strong>Higher initial cost (dedicated degasser unit and vacuum pump).</li></ul><h5><strong>Ease of Use:</strong></h5><ul><li><strong>Offline Degassing:&nbsp;</strong>Manual setup each time which is time-consuming.</li><li><strong>Inline Degassing:&nbsp;</strong>Automated after installation.</li></ul><h5><strong>Impact on Solvent Composition:</strong></h5><ul><li><strong>Offline Degassing:&nbsp;</strong>Some loss of volatiles is possible such as evaporation during sparging.</li><li><strong>Inline Degassing:&nbsp;</strong>Solvent composition remains unchanged.</li></ul><h5><strong>Effect on Chromatographic Stability:</strong></h5><ul><li><strong>Offline Degassing:&nbsp;</strong>Improves initial stability but may allow bubble re-formation later.</li><li><strong>Inline Degassing:&nbsp;</strong>Very stable baselines and retention. Bubble-related problems are virtually eliminated.</li></ul><div><div><div><div><h3><strong>When to Choose Offline vs Inline Degassing</strong></h3></div></div></div></div><div><p>The choice between an offline and inline HPLC degasser depends on the situation, including factors like routine use, troubleshooting needs, and method sensitivity.</p><ol><li>Offline degassing is suitable for occasional use, limited equipment setups, or when routine inline degassing is not available. While inline degassing is preferred for routine analyses requiring consistent performance and high sensitivity.</li><li>Inline degassers have a higher initial cost but save labor. An offline degasser uses low-cost labware and is cheap to set up, though helium or other gases add to the consumable cost.</li><li>If mobile phases contain volatile organics or labile salts, avoid aggressive offline sparging/vacuum (which can evaporate components). Inline vacuum degassing is gentler on the solvent composition.</li><li>Oxygen in the mobile phase is particularly problematic for sensitive methods, such as low-wavelength UV or reductive electrochemical detection. While techniques like helium sparging and inline degassing effectively remove oxygen, remember that offline-sparged solvents are susceptible to reabsorbing air over time, which can lead to signal loss and subtle changes in sensitivity.</li></ol></div><div><div><div><div><h3><strong>Troubleshooting Degassing Issues in HPLC</strong></h3></div></div></div></div><div><h4><strong>1) Cycling of output (pressure or baseline)</strong></h4><p>Cycling patterns in pressure or the chromatogram baseline often indicate poor compressibility at the pump heads. This may result from insufficient degassing efficiency, where adding a second degassing chamber can help. It can also cause air to enter the system due to improperly connected tubing to or from the HPLC solvent degasser. Also, improper connections of the tubing on the HPLC solvent degasser can also allow air to enter the system, causing fluctuations in pressure or baseline.</p><h4><strong>2) Baseline noise</strong></h4><p>Noise on the baseline of a chromatogram may indicate problems with the gas flow system. High gas flow rates can cause short-term noise, while baseline drift (long-term fluctuations) may be due to contaminants being flushed out from a partially clogged gas line. Additionally, an unstable gas flow output can also contribute to baseline noise.</p><h4><strong>3) Leakages</strong></h4><p>Leaks often occur due to incompatibility between solvents and degasser materials. For instance, tetrahydrofuran (THF) can dry and crack degasser membranes. Additionally, improperly seated tubing connections in or out of the degasser can allow leaks to develop.</p><h4><strong>4) Blockages</strong></h4><p>Blockages create pressure ripples as the pump struggles to draw solvent, sometimes causing cavitation. They may result from algae growth, crystallization of buffer salts, or clogged inlet filters. Troubleshooting involves disconnecting outlet tubing to check for continuous solvent flow, while prevention requires filtering solvents, using clean solutions, and flushing lines regularly, especially after system shutdowns.</p><h4><strong>5) General considerations</strong></h4><p>Degasser issues are often linked with the mobile phase quality. Using poorly prepared or contaminated solvents can lead to persistent problems. Maintaining clean, filtered, and fresh solvents helps ensure smooth operation and minimizes downtime.</p></div></div>

Degassing Techniques in HPLC: Inline vs Offline

Everything to know about osmolality

<p><strong>Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 13th July 2026? Check out new documents from the field of liquid phase, especially HPLC and LC/MS techniques!</strong></p><blockquote><p><span style="font-size:20px;">👉 </span><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library"><span style="font-size:20px;"><strong>SEARCH THE LARGEST REPOSITORY OF DOCUMENTS ABOUT LCMS AND RELATED TECHNIQUES</strong></span></a></p></blockquote><blockquote><p><strong>👉 Need info about different analytical techniques?</strong> Peek into <a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/library"><strong>LabRulezGCMS</strong></a> or <a target="_blank" rel="noopener noreferrer" href="https://icpms.labrulez.com/library"><strong>LabRulezICPMS</strong></a> libraries.</p></blockquote><p>This week we bring you posters by <strong>Agilent Technologies / ASMS, Shimadzu / ASMS, Thermo Fisher Scientific / HPLC Symposium </strong>and<strong> Waters Corporation / HPLC Symposium </strong>and technical note by<strong> KNAUER!</strong></p><h3><strong>1. Agilent Technologies / ASMS: Intact Protein Analysis in Plasma Using Automated 2D-LC/TOF with Multiple Heart-Cutting</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_intact_protein_mhc_2d_lc_tof_wp_679_asms_2026_en_agilent_edb54f4854.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Direct intact and top-down protein analysis enables comprehensive characterization of proteoforms, post-translational modifications, and fragments that are often lost in bottom-up workflows.1 However, intact mass analysis in complex biological matrices such as whole plasma is challenged by extreme dynamic range and ion suppression, commonly necessitating affinity-based enrichment.&nbsp;</p><p>Here, we demonstrate a non-affinity, orthogonal separation strategy using a 2D-LC/TOF workflow with multi-heart-cutting for native intact protein analysis directly in plasma. Therapeutic monoclonal antibodies (mAbs) trastuzumab and antibody-drug conjugate T-DM1, were used as model systems to evaluate performance.&nbsp;</p><p>The workflow employs an Agilent 1290 Infinity III 2D-LC system for orthogonal separations and precise multiple heart-cutting, coupled to a high-resolution, high-sensitivity Agilent 6230C Accurate Mass TOF LC/MS System.</p><h4><strong>Experimental</strong></h4><h5><strong>Instrumentation&nbsp;</strong></h5><ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=1290+Infinity+III+2D-LC"><strong>Agilent 1290 Infinity III 2D-LC system </strong></a>comprised the following modules:&nbsp;<ul><li>1290 Infinity III Bio Multisampler with Thermostat (G7137A)&nbsp;</li><li>1290 Infinity III Multicolumn Thermostat (G7116B)&nbsp;</li><li>1290 Infinity III Diode Array Detector (G7117B) with Max-Light cartridge cell (G7117-60020)&nbsp;</li><li>1260 Infinity III Bio Flexible Pump (G7131C)&nbsp;</li><li>1290 Infinity III Bio High-speed Pump (G7132A)&nbsp;</li><li>1290 Infinity Valve Drive (G1170A) with 2D-LC ASM Valve (G5643B)&nbsp;</li><li>Two 1290 Infinity III Valve Drives (G1170A) with multiple heart-cutting valves (G5642-64000) equipped with 40 µL Bio loops&nbsp;</li><li>1290 Infinity III Valve Drive (G1170A) with a 2- position/6-port valve (G4231A) (used as a diverter valve)&nbsp;</li></ul></li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=6230C"><strong>Agilent 6230C Accurate Mass TOF LC/MS System</strong></a></li></ul><h5><strong>LC/MS Analysis&nbsp;</strong></h5><p>Data acquisition and analysis were carried out using <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=MassHunter+acquisition"><strong>MassHunter acquisition</strong></a> (12.2), <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=BioConfirm"><strong>BioConfirm</strong></a> (12.1) and <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=MassHunter+Qualitative+Analysis"><strong>MassHunter Qualitative Analysis </strong></a>(13.0) software.</p><h4><strong>Conclusions</strong></h4><ul><li>Agilent 2D-LC enables direct native MS analysis of intact proteins from plasma without affinity-based enrichment.&nbsp;</li><li>UV-guided and multiple heart-cutting simplify 2D-LC method development and provide high-resolution isolation of native proteins from complex biological matrices such as plasma.&nbsp;</li><li>The Agilent 6230C Accurate Mass TOF delivers high sensitivity and excellent mass accuracy, enabling confident intact protein characterization under native conditions.</li></ul><h3><strong>2. KNAUER: Mind the pressure in preparative up-scaling – effects of particle size and solvent composition on back pressure</strong></h3><ul><li>Technical note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/VTN_0051_knauer_mind_the_pressure_in_preparative_up_scaling_effects_of_particle_size_and_solvent_comp_12e49884d3.pdf"><strong>Full PDF for download</strong></a></li></ul><p>The back pressure of a chromatographic system is generated by the system components, the capillaries, the column, the solvent, and the applied flow rate. Each system has a maximum pressure which it can sustain, and therefore the pressure must be monitored as it is a limiting factor during method development. The back pressure generated by a column is dependent on the particle size and its length. The development of a preparative method is ideally performed on an analytical scale and after optimization the method is then scaled up to preparative scale.</p><p>As preparative HPLC systems normally have lower maximum pressures, it is important to keep the generated pressure in the range of the later preparative system.&nbsp;</p><p>The larger the particles of the column material are, the lower will be the generated back pressure. With increasing particle size, the resolution is decreasing at the same time. Depending on the separation task, the resolution can be a critical point, and it is important to find the right compromise between resolution and backpressure. Additionally, larger particles are significantly less price intensive, which is another factor to be considered especially for large scale purifications. Often solvent mixtures of water with acetonitrile, methanol or ethanol are used for reverse phased methods. Depending on the organic solvent composition, the generated back pressure differs significantly which could be reduced by changing the solvent composition slightly or the organic solvent if possible. The back pressure generated by the solvent will not or only slightly change during the up-scaling process.&nbsp;</p><p>Here, the influence of particle size, flow rate and solvent composition on the back pressure was evaluated. Further, a practical scale-up was performed to evaluate the values obtained on analytical scale to those in larger scale.</p><h4><strong>User Tips:&nbsp;</strong></h4><ul><li>What is the finale scale you want to operate later?&nbsp;</li><li>What are the pressure limitations of the preparative system?&nbsp;</li><li>Be aware of the solvent back pressure&nbsp;</li><li>Consider particle size not only for best separation&nbsp;</li><li>Consider the flow rate</li></ul><h4><strong>MATERIAL AND METHODS&nbsp;</strong></h4><ul><li><strong>Pump: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/2372"><strong>AZURA® P 6.1LHPG</strong></a>, 10 ml, sst&nbsp;</li><li><strong>Autosampler: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/2375"><strong>AZURA AS 6.1L</strong></a></li><li><strong>Detector:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/2423"><strong>UV AZURA DAD 2.1L&nbsp;</strong></a></li><li><strong>Flow cell: </strong>Pressure proof 10 mm, 10 µl, 300 bar&nbsp;</li><li><strong>Thermostat:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/2376"><strong>CT2.1&nbsp;</strong></a></li><li><strong>Columns:&nbsp;</strong><ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1633"><strong>Eurospher II 100-5 C18, 150 x 4.6 mm&nbsp;</strong></a></li><li>Eurospher II 100-10 C18, 150 x 4.6 mm&nbsp;</li><li>Eurospher II 100-15 C18, 150 x 4.6 mm&nbsp;</li><li>Eurospher II 100-20/45 C18, 150 x 4.6 mm &nbsp;</li></ul></li><li><strong>Software&nbsp;</strong><ul><li>ClarityChrom 10.1&nbsp;</li><li>ClarityChrom 10.1 PDA extension</li></ul></li></ul><h4><strong>CONCLUSION&nbsp;</strong></h4><p>The back pressures of four C18 150 x 4.6 mm columns, which differed only in particle size were measured and compared at different flow rates and solvent compositions. As expected, the 5 µm column showed the highest and the 20/45 µm column the lowest back pressure at all tested conditions. The pressure curves of ethanol and methanol gradients were similar, with a maximum pressure at approximately 50% solvent. The pressure curve of the acetonitrile gradient differed from the other two, showing the highest pressure at a lower organic solvent content of approximately 35%, after which the pressure dropped. Ethanol showed the highest pressure, followed by methanol and then acetonitrile. Determining the column back pressure at an analytical scale is an important tool during preparative method development. Ideally, if a linear scale up from the analytical to the preparative scale is conducted, the column back pressure should stay the same. This helps to predict if the method can be scaled up to the desired size in terms of back pressure.</p><h3><strong>3. Shimadzu / ASMS: Enhanced Quantification of Emerging UV Filter Contaminants in Drinking Water Using Optimized LC-MS/MS Technique</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/TP_121_Enhanced_Quantification_of_Emerging_UV_Filter_Contaminants_in_Drinking_Water_Using_Optimized_LC_MSMS_Technique_e733c64828.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Ultraviolet (UV) filter chemicals from sunscreens and personal care products are increasingly recognized as emerging contaminants of concern in drinking water1 . Their persistence and reported biological effects highlight the need for sensitive and reliable monitoring approaches2 . The objectives were to optimize quantitation to achieve ppt level detection without offline extraction and assess the occurrence of these compounds in multiple drinking water matrices. In this study, 20 UV filters were analyzed using a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=LCMS-8060RX"><strong>Shimadzu LCMS8060RX triple quadrupole mass spectrometer.</strong></a></p><h4><strong>Conclusion</strong></h4><p>A highly sensitive, precise, and robust direct injection LC–MS/MS method enables sub-ppt to high-ppb quantification of 20 UV filters in drinking water. Overall, the highly sensitive and robust Shimadzu LCMS-8060RX Triple Quadrupole Mass Spectrometer combined with this extraction-free workflow provides a simple, reliable, and high-throughput approach for quantifying emerging UV filters at ng/L concentrations in drinking water.</p><h3><strong>4. Thermo Fisher Scientific / HPLC Symposium: Comparison of biocompatible and inert UHPLC systems for LC-UV quantitation of large RNA</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_004752_hplc_vanquish_amplify_metal_sensitive_analyte_analysis_po004752_na_en_ff99cfde49.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Accurate LC-UV analysis of large RNA and oligonucleotide therapeutics requires highly inert chromatographic flow paths because interactions between the negatively charged phosphodiester backbone and metallic surfaces can cause analyte loss, peak broadening, and reduced quantitative sensitivity, particularly at low concentration.&nbsp;</p><p>Here, an inert UHPLC system – Vanquish Amplify system, incorporating inert-coated flow path components was evaluated against a biocompatible UHPLC configuration using MP35N alloy capillaries and additionally against a commercially available inert UHPLC platform. Large RNA fragments of 500 and 5000 nucleotides were analyzed by IP-RPLC to assess the difference between these systems. To further investigate flow path interactions, adenosine 5′- (α,β-methylene) diphosphate (AMPcP), a metal-sensitive analyte, was analyzed alongside adenosine, a noninteracting reference compound.</p><h5><strong>Material and Methods</strong></h5><ul><li><strong>Instrumentation: </strong>Thermo Scientific Vanquish Amplify UHPLC system</li><li><strong>Column:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=3&amp;search=SurePac+Oligo"><strong>Thermo Scientific SurePac Oligo RP MDi HPLC column</strong></a>,<br>2.5 µm 2.1 X 50 mm (P/N: 43712-052132</li><li><strong>Data Analysis: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/76"><strong>The Thermo Scientific Chromeleon Software 7.3.2 MUc Chromatography Data System (CDS)</strong></a> was used for all data acquisition and analysis.</li></ul><h4><strong>Conclusions&nbsp;</strong></h4><p>Using AMPcP with the Vanquish Amplify system demonstrated up to a twofold increase in sensitivity compared to systems with MP35N fluidics under conditions where metal interactions are most pronounced&nbsp;</p><p>For large RNA, the Vanquish Amplify UHPLC system delivered 1.25–1.35× higher signal intensity compared to systems with MP35N fluidics, enabling reduced sample consumption and improved quantitative reliability.&nbsp;</p><p>The Vanquish Amplify UHPLC system demonstrated comparable system inertness under the tested conditions for larger RNA fragments (5,000 nt) and slightly better system inertness for smaller fragments (500 nt) in this study than the MaxPeak HPS containing system, based on peak height and area comparisons using an external UV detector</p><h3><strong>5. Waters Corporation / HPLC Symposium: Development and Transfer of Quantitative Analytical Methods using Universal Detectors</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/720009493_8cbe4c3cc2.pdf"><strong>Full PDF for download</strong></a></li></ul><div><div><section><div><div><div><div><div><div><p>Universal detectors such as <strong>Charged Aerosol Detection (CAD)</strong> and <strong>Evaporative Light Scattering Detection (ELSD)</strong> provide valuable alternatives to traditional LC detectors including UV, fluorescence, and mass spectrometry, particularly for analytes lacking chromophores. Although both detectors are suitable for quantitative analysis, they rely on different detection principles—CAD measures the electrical charge transferred to aerosol particles, whereas ELSD detects light scattered by dried analyte particles. Because both detectors exhibit non-linear responses, quantitative methods typically require calibration models such as log-log linear or quadratic regression rather than conventional linear calibration. The poster discusses key considerations for developing robust quantitative methods and highlights the challenges associated with transferring methods between different universal detection platforms.</p><p>To demonstrate detector transferability, the authors migrated a <strong>HILIC method for sugar analysis in fruit juice</strong> from ELSD to CAD. The separation was performed on a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/306"><strong>Waters Arc HPLC System</strong></a> equipped with an <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=5&amp;search=XBridge+BEH+Amide"><strong>XBridge BEH Amide</strong></a><strong> (4.6 × 150 mm, 2.5 μm)</strong> column using a water/acetonitrile gradient containing triethylamine. While the chromatographic conditions remained unchanged, CAD detector parameters were optimized. Both detectors produced highly comparable quantitative performance across a calibration range of <strong>50–1000 μg/mL</strong>, achieving calibration coefficients (R²) above <strong>0.995</strong>, area RSD values below <strong>1.5%</strong>, and nearly identical concentrations of fructose, sorbitol, glucose, and sucrose in an apple juice sample. These results demonstrate that quantitative HILIC methods can be successfully transferred between ELSD and CAD with only detector-specific optimization.</p><p>The poster also evaluates method transfer between different CAD platforms using the <strong>USP Deoxycholic Acid assay</strong>. The method was implemented on a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/198"><strong>Waters ACQUITY UPLC H-Class PLUS System</strong></a> with an <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1823"><strong>XBridge BEH C18</strong></a><strong> (4.6 × 150 mm, 3.5 μm)</strong> column and compared against a competing LC/CAD system. After scaling the method according to <strong>USP Chapter &lt;621&gt;</strong>, both systems met all USP system suitability requirements, including repeatability and signal-to-noise criteria. Quantitative results for deoxycholic acid and related impurities were virtually identical, confirming that validated CAD methods can be reliably transferred between different vendors' LC/CAD platforms without compromising analytical performance.</p><p>Overall, the study demonstrates that both ELSD and CAD are robust universal detection techniques for quantitative liquid chromatography. While method transfer between identical detector types is generally straightforward, successful migration between ELSD and CAD—or between different CAD systems—requires careful optimization of detector parameters while preserving chromatographic conditions. The presented workflows illustrate that with appropriate calibration and detector tuning, laboratories can achieve equivalent sensitivity, precision, and quantitative accuracy, facilitating method standardization and transfer across analytical platforms.</p></div></div></div></div></div></div></section></div></div>

News from LabRulezLCMS Library - Week 29, 2026

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

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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.

Phenomenex

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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Tosoh Bioscience

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Waters Corporation

<p><strong>An online Thermo Scientific Proteome Discoverer crosslinking training workshop delivered via Microsoft Teams, designed to get novice/intermediate users up to speed with the PD 3.0 (SP1) software package.</strong></p>
<p>Two interactive on-line sessions, featuring hands-on exercises, discussion of features, parameters and optimised settings.</p>


Thermo Scientific: Proteome Discoverer XlinkX Workshop

Proteome Discoverer XlinkX Workshop

An online Proteome Discoverer crosslinking training workshop delivered, designed to get novice/intermediate users up to speed with the PD 3.0

<p><strong>CHROMATOGRAPHY OF BIOSEPARATIONS 101 WEBINAR SERIES</strong></p>
<p><strong>This webinar series will cover the foundations of chromatography for the separation and analysis of peptides and proteins.  Understanding the fundamentals of bioseparations will enable you to develop methods more efficiently.  At the end of this series, you will be up to date on cutting edge principles and innovations surrounding bioseparations.</strong></p>
<p>Who should attend? </p>
<ul>
<li>Those interested in a refresher on foundational principles and tips for bioseparations  </li>
<li>Those revisiting which columns and mobile phases are optimal for which workflows </li>
<li>Those looking for an overview of industry advancements and new offerings</li>
</ul>
<blockquote>
<p><strong>Presenter:</strong> Bill Warren (Principal Product Manager – Chemistry Technology Center, Waters Corporation)</p>
</blockquote>
<blockquote>
<p>Bill received his Masters in Microbiology and Immunology from University of Louisville School of Medicine. In 1986, he joined Waters Corporation as a senior applications chemist involved with the research and development of HPLC, FPLC, and Capillary Electrophoresis applications for the analysis and purification of proteins, peptides, and nucleic acids. Following four years as a field based technical sales representative for Waters, Bill enjoyed a tenure as Manager, Core Product Evaluation Laboratory within the Waters Worldwide Marketing Organization. Bill currently is a Principal Bioseparation Columns Product Manager whose responsibilities involve assisting in the development and commercialization of innovative reagents and chromatography-based technologies for the separation, purification, and characterization of proteins and peptides.</p>
</blockquote>
<blockquote>
<p><strong>HOST:</strong> Zerene Manaloto (Chemistry Products Marketing Manager, Waters Corporation)</p>
</blockquote>
<blockquote>
<p>Zerene is a Chemistry Product Marketing Manager within the Americas Field Marketing team. She holds a Chemical Engineering Bachelor’s degree from Stevens Institute of Technology. Early in her career, Zerene was a research scientist at Bristol-Myers Squibb in Process Research &amp; Development involved in preparative scale purification and glass plant operations to support IND and clinical supplies of drug candidates. Prior to joining Waters, Zerene was in consumable field sales, supporting key accounts and customers within the pharmaceutical and clinical industries.</p>
</blockquote>


Waters Corporation: CHROMATOGRAPHY OF BIOSEPARATIONS 101 WEBINAR SERIES

CHROMATOGRAPHY CONCEPTS FOR PEPTIDES AND PROTEINS

This webinar series will cover the foundations of chromatography for the separation and analysis of peptides and proteins.

CHROMATOGRAPHY CONCEPTS FOR PEPTIDES AND PROTEINS 

<p><strong>Let your polar pesticides be our problem!</strong></p>
<p><strong>Polar pesticides, the most widely used group of agrochemicals in the world, are few of the most challenging compounds to be determined by analytical methods. So, laboratories, tasked with detecting and characterizing polar pesticides in diverse samples, had to work with complex individual or group methods with time-consuming derivatizations.</strong></p>
<p>This is exactly what we aim to address through this exclusive Deminar “Let’s Talk Polar Pesticides”.</p>
<p>During this Deminar, our speakers will take you through the various aspects of the Polar pesticides problem, from regulatory guidelines to workflow solutions, live from Agilent Lab.</p>
<p>The topics address the following aspects for the Analysis of Polar Pesticides in Food:</p>
<ul>
<li><strong>FSSAI/EU guidelines and DG/SANTE</strong></li>
<li><strong>Specific analytical challenges to the analysis of Polar pesticides</strong></li>
<li><strong>Detailed workflow solution using Agilent LC-MS/MS</strong></li>
<li><strong>Sensitivity and matrix-based linearity in accordance with EU norms</strong></li>
<li><strong>Virtual Demo from Agilent Lab including actual sample preparation (with recovery experiments at LOQ level) and the final data as acquired in Agilent COE on actual samples</strong></li>
</ul>
<p>What You will Learn</p>
<ul>
<li>LCMS based analysis of Polar Pesticides without derivatization.</li>
<li>Learn how to adreess peak tailing and sensitivity issue</li>
<li>Analysis of difficult food matrices such as cereals and tea.</li>
</ul>
<p>Who should attend?</p>
<p>Lab directors, managers and mass spectrometry analysts performing small molecule analysis in food </p>
<blockquote>
<p><strong>Presenter:</strong> Vikrant Goel (Application Scientist, Agilent Technologies)</p>
</blockquote>
<blockquote>
<p>Vikrant comes with an extensive 19 years of experience working in QA/QC, R&amp;D and Analytical roles and an expert in various LCMS technologies. His solution-based approach has been well accepted among LCMS fraternity from Pharma and Applied market segments, since 2004. He holds an M.tech in Biotechnology, PG Dip in Bioinformatics &amp; B. Pharm. He joined Agilent in 2010, as a Product specialist for LC/MS, after having worked with ILS Bioservices &amp; LabIndia Instruments. His passion towards solution, brought him back to the Application role, 6 years ago.</p>
</blockquote>


Agilent Technologies: Let's Talk Polar Pesticides

Let's Talk Polar Pesticides

Our speakers will take you through the various aspects of the Polar pesticides problem, from regulatory guidelines to workflow solutions, live from Agilent Lab.

<ul>
<li><p><strong>Receive a demonstration of how non-denaturing SEC-Native MS can be leveraged to streamline mAb characterization, including methods for the digestion of mAbs</strong></p>
</li>
<li><p><strong>Listen in to a column selection discussion, which includes pore volume and column dimensions to ensure monomer and aggregate resolution, and optimize mobile phases for complex characterizations</strong></p>
</li>
<li><p><strong>Learn about SEC combined with nHRMS, and how to select a SEC column</strong></p>
</li>
</ul>
<p>Who Should Attend:</p>
<p>Any scientists wanting to learn how to combine their size exclusion chromatography and High-resolution Mass Spec for improvements in their mAb characertizations. </p>
<blockquote>
<p><strong>Presenter:</strong> Lorne Nelson (Senior Scientist - Field Applications, Phenomenex)</p>
</blockquote>


Phenomenex: SEC-nHRMS for the Characterization of mAbs

SEC-nHRMS for the Characterization of mAbs

Receive a demonstration of how non-denaturing SEC-Native MS can be leveraged to streamline mAb characterization, including methods for the digestion of mAbs

<p><strong>In this webinar we shall take a look at the contents of the long awaited GAMP®5 Version 2 release to examine what has truly changed since the Version 1 release, with special focus on what might impact regulated laboratories in the Pharma, BioPharma, and Biotech industries.</strong></p>
<p>The audience should learn what the major changes are for Version 2 and how they might impact their current regulated lab operations so they can make pro-active updates to their current compliance related procedures and policies.</p>
<p>By attending this webinar you will:</p>
<ul>
<li><strong>Understand which sections of GAMP®5 Version 2 have been updated since Version 1.</strong></li>
<li><strong>Understand which updated sections may have an impact on your current business processes.</strong></li>
<li><strong>Understand the trends of the industry regarding the evolution of compliance activities related to evolving technologies and business practices.</strong></li>
</ul>
<p>Who should attend:</p>
<ul>
<li>Lab Managers</li>
<li>Lab SMEs</li>
<li>Validation Engineers</li>
<li>IT Professionals engaged in Validation/regulated activities</li>
<li>QA professionals</li>
<li>Senior Lab Management</li>
</ul>
<blockquote>
<p><strong>Presenter:</strong> Michael Korbel (Global Sr. Product Manager, Agilent Technologies, Inc.)</p>
</blockquote>
<blockquote>
<p>Michael has 25 years of experience in regulated environments in the Pharmaceutical and Bio-Tech industries. Half of Michael&#39;s career has been spent on the customer side of the industry in both IT and Laboratory positions; supporting, qualifying, and validating regulated computerized systems and instruments. The other half has been spent providing consultative compliance services from the vendor side of the industry. His experience includes validating laboratory informatics systems, as well as instrument qualification, preventive maintenance, and relocation services.</p>
</blockquote>
<blockquote>
<p>Michael has held various IT and Lab positions at MannKind Corporation and Purdue Pharma, as well as consulted for Pfizer, JnJ, TEVA, and Merck. Michael has also held various positions at Waters Corporation, PerkinElmer OneSource, and Agilent Technologies. He currently serves as the Senior Product Manager of Compliance Consulting services on the global Marketing team at Agilent Technologies. Michael holds a B.S. in Biology, with minors in Chemistry and Environmental Science.</p>
</blockquote>


Pixabay/Gerd Altmann: Computerized System Validation: GAMP 5 2nd Edition, What You Need to Know

Computerized System Validation: GAMP 5 2nd Edition, What You Need to Know

We shall take a look at the contents of the long awaited GAMP 5 Version 2 release to examine what has truly changed since the Version 1 release...

System Validation: GAMP 5 2nd Edition, What You Need to Know

<p>Key learning objectives</p>
<ul>
<li><p><strong>Guidance to making Adjustments to chromatographic methods</strong></p>
</li>
<li><p><strong>Column Phases, Particle Sizes, Internal diameter, Flow rate</strong></p>
</li>
<li><p><strong>Composition, Injection volume, Dwell volume</strong></p>
</li>
<li><p><strong>System suitability</strong></p>
</li>
</ul>
<blockquote>
<p><strong>Presenter:</strong> Dr.Heiko Behr (Senior Business Development Manager, Europe)</p>
</blockquote>


Phenomenex: Harmonisation of Ph.Eur. 2.2.46 and USP 621

Harmonisation of Ph.Eur. 2.2.46 and USP 621

Guidance to making Adjustments to chromatographic methods...

<p>Key learning objectives</p>
<ul>
<li><p><strong>Reversed Phase-Ion Pair LC for Oligos - Challenges and Optimization</strong></p>
</li>
<li><p><strong>Rapid Isolation of Oligo Therapeutics from Biological Samples</strong></p>
</li>
<li><p><strong>High-throughput, RPC Purification</strong></p>
</li>
</ul>
<blockquote>
<p><strong>Presenter:</strong> Mahesh Thondawada (Business Development Manager Biopharma/Biologic)</p>
</blockquote>


Analytical BioSolutions for Therapeutic Oligonucleotides

Reversed Phase-Ion Pair LC for Oligos - Challenges and Optimization...

<p><strong>In today&#39;s fast-paced digital world, the ability to make data-driven decisions rapidly is pivotal to the laboratories, in order to boost productivity, shorten response time, release analysis results quickly, and plan for future expansions. However, as crucial as this need may be, it has been especially difficult in many QC and research laboratories due to the lack of a system that provides a synopsis of Lab Performance statistically.</strong></p>
<p>In this webinar, you will witness the power of <strong>LabSolutions BiX</strong> - <strong>the comprehensive analytics platform</strong> - which can provide users with accurate and timely laboratory analytics, from a laboratory, at home, or from a remote location around the world. Whether you are using Chromatography, Spectroscopy, or Mass Spectrometry instruments, LabSolutions BiX possess the capability to capture, process, and analyze all the data from respective software, including Shimadzu&#39;s unique network platform LabSolutions CS, and showcase the analyzed output into various Analytics dashboard reports.</p>
<p>Join us to learn how you can achieve &quot;Lab-at-a-glance&quot; easily using LabSolutions BiX, which enables the monitoring of the Live Status of laboratory activities, the efficiency of laboratory users, instrument utilization, and other new features in this latest version which will be unveiled during the webinar.</p>
<p>Learning Objectives:</p>
<ul>
<li><strong>How to get the insights of Lab Perfomance in graphical visualization with Live Dashboard</strong></li>
<li><strong>How to monitor Instrument utilization and user performance to enhance the productivity</strong></li>
<li><strong>Familiarize on “Audit Dashboard” for quick review &amp; release of batches</strong></li>
<li><strong>Understand how to leverage LabSolutions BiX unique features for your laboratory</strong></li>
</ul>
<blockquote>
<p><strong>Presenter:</strong> Dr Kanimozhi Sivarajan (Assistant Informatics Manager, Shimadzu (Asia Pacific) Pte Ltd)</p>
</blockquote>
<blockquote>
<p>Dr Kanimozhi is leading the Informatics sales &amp; marketing division for the APAC region, providing Laboratory Informatics Solutions such as LabSolutions CS, LIMS and other QMS applications. She is supporting customers with implementation of laboratory automation solutions and is responsible for providing consultation on streamlining Lab Workflow, Regulatory Compliance requirements for Electronic Data Management and Lab Practices/Analysis.</p>
</blockquote>
<blockquote>
<p><strong>Guest Speaker:</strong> Mr Akbar Kamal (CIO - Digital Transformation &amp; Data Analytics, Zim Laboratories Ltd)</p>
</blockquote>
<blockquote>
<p>Mr Akbar Kamal is the CIO at Zim Laboratories Ltd. His responsibilities include understanding user requirements for integrating 7 major software systems relating to various business systems for quicker &amp; better interpretation of data resulting in better inter-company decision making. He works with Hybrid Cloud (Azure) &amp; Chromium-based HDMI dashboards for automated reporting and has good hands on coding experience with C, SQL &amp; Automation Scripting.</p>
</blockquote>


Shimadzu: Introducing the latest version Lab-at-a-glance powered by LabSolutions BiX

Introducing the latest version Lab-at-a-glance powered by LabSolutions BiX

LabSolutions BiX enables the monitoring of the Live Status of laboratory activities, the efficiency of laboratory users, instrument utilization ...

The latest version Lab-at-a-glance by LabSolutions BiX

<p><strong>Design–build–test–learn (DBTL) cycles are commonly used in biotechnology and in natural product discovery as they provide a systemic and efficient framework for the metabolic engineering of microbes.</strong></p>
<p>A novel platform that automates multiomics analyses of microbes could enable faster and better DBLT cycles for your research.</p>
<p>In this webinar, the development and validation of the high-throughput, automated platform using model microbial organisms is described, as well as the recent work in applying the platform to cell factory and natural products engineering.</p>
<p>Attend this webinar to:</p>
<ul>
<li><strong>Learn how automation can enable better and faster DBLT cycles in the life sciences</strong></li>
<li><strong>Evaluate the pros and cons of taking an automated and digital approach to life science workflows</strong></li>
<li><strong>Discover the next frontiers of technology that will have an impact in biotechnology</strong></li>
</ul>
<blockquote>
<p><strong>Presenter:</strong> Dr. Douglas McCloskey (Independent Consultant )</p>
</blockquote>


Technology Networks: Advances in Multiomics Screening of Microbes in Biotechnology

Advances in Multiomics Screening of Microbes in Biotechnology

In this webinar, the development and validation of the high-throughput, automated platform using model microbial organisms is described.

Multiomics Screening of Microbes in Biotechnology

<p><strong>Christian Rohrer, Director Sales and Marketing at Tosoh Bioscience GmbH, will present the next edition of Tosoh Tuesday, a webinar series held by Chromatography Experts.</strong></p>
<p>On Tuesday, February 14, Andrea Krumm, Product Manager Analytical Columns at Tosoh Bioscience, will present how powerful Hydrophobic Interaction Chromatography is in analyzing proteins.</p>
<p>You will learn how:</p>
<ul>
<li><p><strong>HIC works</strong></p>
</li>
<li><p><strong>HIC methods are developed</strong></p>
</li>
<li><p><strong>HIC is applied to analyze various protein modifications</strong></p>
</li>
</ul>
<blockquote>
<p><strong>Presenter:</strong> Christian Rohrer (Director Sales and Marketing at Tosoh Bioscience GmbH)</p>
</blockquote>


Analytical BioSolutions for Therapeutic Oligonucleotides

Computerized System Validation: GAMP 5 2nd Edition, What You Need to Know

Proteome Discoverer XlinkX Workshop

Advances in Multiomics Screening of Microbes in Biotechnology

HIC for protein analysis: why, what, and how?

Introducing the latest version Lab-at-a-glance powered by LabSolutions BiX

CHROMATOGRAPHY CONCEPTS FOR PEPTIDES AND PROTEINS

Let's Talk Polar Pesticides

Harmonisation of Ph.Eur. 2.2.46 and USP 621

SEC-nHRMS for the Characterization of mAbs

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