Solving the retention time repeatability problem of hydrophilic interaction liquid chromatography

Profiling of free fatty acids in wort using an isolator column-assisted LC-MS/MS approach

News from LabRulezLCMS Library - Week 10, 2026

<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

<p><strong>Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 20th 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 / ASMS </strong>and technical note by<strong> KNAUER!</strong></p><h3><strong>1. Agilent Technologies / ASMS: Comprehensive characterization of oligonucleotides and related impurities using advanced LC/Q-TOF analytical workflows</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_oligonucleotide_lc_q_tof_mp_536_asms_2026_en_agilent_b896ffee63.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Therapeutic oligonucleotides are synthetic, short nucleic acid sequences designed to modulate gene expression by targeting RNA or DNA with high sequence specificity. However, their inherent structural heterogeneity makes impurity profiling essential, as during GMP manufacturing, impurities like phosphorothioate to phosphodiester, 5′ and 3′ truncations or extensions, depurination, and abasic species can directly impact both product safety and efficacy. Antisense oligonucleotides (ASO) like Nusinersen (7127.171 g/mol) are indicated for the treatment of spinal muscular atrophy (SMA). Their closely related variants coelute with the main peak, complicating their characterization in conventional one-dimensional LC/MS workflows. To address this challenge, a 2D LC/MS strategy was implemented using the Agilent 1290 Infinity III Bio LC system with multiple heart cutting, coupled to the 6545XT AdvanceBio LC/Q-TOF and MassHunter BioConfirm software. The first-dimension separation uses an ion pairing buffer-based method to achieve higher chromatographic peak resolution, while the second dimension provides MS compatible desalting, enabling enhanced mass spectrometric sensitivity and improved detection of low-level impurities.&nbsp;</p><h4><strong>Instrumentation</strong></h4><p>1290 Infinity III Bio 2D-LC &amp; <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=6545XT"><strong>6545XT Q-TOF</strong></a> includes:&nbsp;</p><ul><li>1290 Infinity III Bio Flexible Pump (G7131A)&nbsp;</li><li>1290 Infinity III Bio High-Speed Pump (G7132A)&nbsp;</li><li>1290 Infinity III Bio Multisampler (G7137A)&nbsp;</li><li>1290 Infinity III Multicolumn Thermostat (G7116B)&nbsp;</li><li>1290 Infinity III Variable Wavelength Detector (G7114B)&nbsp;</li><li>1290 Infinity III Diode Array Detector (G7117B)&nbsp;</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=2D-LC"><strong>2D-LC System</strong></a> with MHC set up</li></ul><h4><strong>Conclusions</strong></h4><ul><li>A 2D-LC/MS workflow enabled effective separation and identification of coeluting impurities.&nbsp;</li><li>HiRes sampling with multi-injection improved resolution and throughput while preserving 1D separation for MScompatible desalting.&nbsp;</li><li>6545XT AdvanceBio LC/Q-TOF delivered accurate mass and MS/MS data for low-abundance impurity detection.&nbsp;</li><li>MassHunter BioConfirm software enabled automated sequence confirmation and impurity identification.&nbsp;</li><li>This integrated platform offers a robust solution for oligonucleotide characterization in therapeutic development.</li></ul><h3><strong>2. KNAUER: Smart Monitoring &amp; Optimization Solutions for SMB Processes: In-Line and Off-Line Tools for SMB Systems</strong></h3><ul><li>Technical note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/VTN_0048_knauer_in_line_and_off_line_tools_for_smb_systems_50a5f1c1cf.pdf"><strong>Full PDF for download</strong></a></li></ul><p>In liquid chromatography, the purification of target compounds is classically carried out using single column batch chromatography. More efficient approaches include semi – continuous systems with two to three columns and continuous multicolumn systems. The most widely used of the latter is the simulated moving bed (SMB) chromatography which enables the continuous separation of binary or pseudo-binary mixtures. SMB purification is used in various areas such as petrochemical, pharmaceutical or food industry for the production of fine chemicals in different quantities and purities ranging from gram to tons per day. SMB systems vary significantly in size from small lab scale systems operating columns with 8 mm inner diameter (ID) to industrial systems operating columns of onemeter ID or larger.</p><p>Ideally, an SMB process is developed in a smaller lab scale system to reduce solvent and sample consumption compared to development at production scale. Once the separation parameters are established, the SMB process allows for accurate up scaling from lab to production scale. During method development but also production, different tools can be used to monitor and control the process which can be distinguished between in-line and off-line measures. The most crucial parameter, in addition to measuring the pressure, is monitoring the flow rates of the pumps, outlets and/or zones accurately with additional flowmeters. Normally, SMB systems are not equipped with detectors, but UV or RI detectors can be integrated to monitor process stability.&nbsp;</p><p>A standard off-line tool is used to analyse samples taken for one switch or cycle at the raffinate and extract outlets. The resulting chromatograms are used to check, if the separation is working. Further, samples can be taken during the process allowing the depiction of concentration profiles in the different zones. These tools will be described in more detail in the following sections.</p><h4><strong>MATERIAL AND METHODS</strong></h4><ul><li><strong>AZURA Lab SMB system</strong>: SMB, biocompatible, 30 ml/min zone1, 4 ml/min feed, 10 – 130 bar&nbsp;</li><li><strong>AZURA Lab SMB system: </strong>SMB, stainless steel, 30 ml/min zone1, 4 ml/min feed, 10 – 130 bar&nbsp;</li><li><strong>AZURA Pilot SMB system, standard: </strong>SMB, stainless steel, 250 ml/min zone1, 40 ml/min feed, 2 – 100 bar&nbsp;</li><li><strong>AZURA Pilot SMB system, high flow:</strong> SMB, stainless steel, 400 ml/min zone1, 100 ml/min feed, 2 – 60 bar</li><li><strong>Refractive index detector:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/2429"><strong>AZURA® RID2.1L HighFlow detector</strong></a> 100 ml/min&nbsp;</li><li><strong>UV detector: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/2428"><strong>AZURA®UVD 2.1S detector</strong></a>, w/o flow cell</li><li><strong>UV detector: </strong>AZURA®UVD2.1S UV detector, fiber optics w/o flow cell</li></ul><h4><strong>CONCLUSION&nbsp;</strong></h4><p>SMB is a continuous separation process that can run for several hours or days without interruption. Therefore, it is important to monitor the process and to be able to adapt to changes during the run. Different tools are available that can be used for process monitoring and method development. These tools can be divided into two main categories: in-line and off-line tools. In-line tools monitor parameters such as pressure and flow rate directly during the run, whereas off-line tools take a sample, which is then analyzed by an HPLC system. The tools described can be added to the KNAUER SMB systems depending on the customer’s needs. The more parameters that are measured, the better the process can be monitored. This is especially helpful during SMB method development.</p><h3><strong>3. Shimadzu / ASMS: Accelerating drug screening in forensic hair analysis by applying high-speed polarity switching in HR LC-MS/MS</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/TP_180_Accelerating_drug_screening_in_forensic_hair_analysis_by_applying_high_speed_polarity_switching_in_HR_LC_MSMS_168f8cc0b2.pdf"><strong>Full PDF for download</strong></a></li></ul><p>In 2023, the Society of Hair Testing (SoHT) published updated consensus guidelines for the forensic analysis of drugs of abuse in hair, defining recommended targets across eight major drug classes: opiates, cocaine, amphetamines, cannabinoids, opioids, methadone, buprenorphine, and ketamine. Legal drivers, accepted use cases, and degree of prescription differ between US and Europe but in both jurisdictions, mass spectrometry is widely applied to support legally defensible conclusions. The challenge for any drug testing in hair samples, whether it is in forensic and safeguarding contexts, workplace testing or a judicial setting is the need to develop methods capable of detecting expanded search panels with the increased prevalence of new psychoactive substances (NPS) and non-medical prescription drug use. The approach applied in this work considered a LC/QTOF analysis using DDAMS/MS with rapid polarity switching with a novel data processing application, Insight Profiler. The software was designed to support a single click approach to automate data processing of complex data sets in non-targeted analysis from feature detection to compound identification.</p><h4><strong>Materials and Methods</strong></h4><h5><strong>Reversed phase LC Separation: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=Nexera+X2"><strong>Nexera X2&nbsp;</strong></a></h5><ul><li>Shim-pack Velox Biphenyl (100 x 2.1 mm, 2.7 µm) column; column temp. 40 ºC, flow rate: 0.3 mL/min, 17 min total analysis time. Methanol:water gradient with 2 mM ammonium formate and 0.002% formic acid&nbsp;</li></ul><h5><strong>High resolution QTOF analysis: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=LCMS-9050"><strong>LCMS-9050&nbsp;</strong></a></h5><ul><li>MS scan m/z 100-1000, 100 msec scan time. Polarity switching time 800 msec.&nbsp;</li><li>MS/MS DDA: 4 scans in positive ion mode (m/z 40-1000, 33 msec), 2 scans in negative ion mode (m/z 40-1050, 33 msec) CE 5-55V (1.998 sec total cycle time).</li></ul><h5><strong>Data processing</strong></h5><p><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=Insight+Profiler"><strong>Insight Profiler</strong></a>, feature detection threshold set to low, compound identification search lists included NIST, HighResNPS, Shimadzu Forensic Toxicology database.</p><h4><strong>Conclusions&nbsp;</strong></h4><ul><li>The targeted DDA-MS/MS polarity switching approach successfully identified a significant number of unknown compounds in the samples not routinely targeted by triple quadrupole analysis.&nbsp;</li><li>The method using DDA-MS/MS was easily configured to support an expanded DoA target panel to meet the needs of forensic toxicology screening.&nbsp;</li><li>Non-targeted analysis using Insight Profiler accelerated data analysis by aligning identified compounds from the same hair donor showing direction of change for pharmaceutical and illicit compounds detected.</li></ul><h3><strong>4. Thermo Fisher Scientific / HPLC Symposium: Enhancing semi-volatile analyte detection in HPLC-CAD using temperature coupling mode for optimized evaporation control</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_004766_hplc_vanquish_cad_fatty_acid_quantification_po004766_na_en_4a0868c2cb.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Polysorbates are widely used in biopharmaceutical formulations but can degrade or contain FAs impurities that must be accurately measured to ensure product quality and safety. Charged Aerosol Detection is commonly used for FA analysis, though its performance depends strongly on EvapT. Higher EvapT improves S/N ratios but can reduce sensitivity for semi-volatile compounds like FAs, while lower EvapT has the opposite effect. The <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/2176"><strong>Thermo Scientific Vanquish Charged Aerosol Detector P</strong></a> series introduces a Temperature Coupling Mode that links EvapT with the charging detection module (CDM) temperature, overcoming limitations of earlier systems with fixed CDM temperature of 40 ºC. This coupling improves response and measurement reliability for semi-volatile species, enabling better optimization of FA analysis<sup>1</sup>.</p><h4><strong>Conclusions&nbsp;</strong></h4><ul><li>Advanced temperature control in CAD improved method sensitivity up to ~47% for semi-volatile fatty acids.&nbsp;</li><li>The best overall S/N was achieved at 25.0 °C EvapT with Temperature Coupling Mode enabled.&nbsp;</li><li>The optimized CAD approach supports more sensitive FA impurities and degradants quantification in pharmaceutical and biopharmaceutical formulations.</li></ul><h3><strong>5. Waters Corporation / ASMS: High mass and spatial resolution MS imaging of soft fruit sample cross-sections using desorption electrospray ionization (DESI)</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/720009492_282619e388.pdf"><strong>Full PDF for download</strong></a></li></ul><div><div><section><div><div><div><div><div><div><p>This poster demonstrates the use of <strong>desorption electrospray ionization mass spectrometry imaging (DESI MSI)</strong> for visualizing the spatial distribution of metabolites in high-water-content soft fruits, specifically strawberries and raspberries. The authors developed a simple direct-transfer workflow to overcome the challenges associated with imaging delicate fruit tissues and compared three transfer substrates: plain glass microscope slides, nitrocellulose membranes, and Hamamatsu Poropare plates. The goal was to determine which surface best preserves metabolite localization while enabling high-quality mass spectrometry imaging.</p><p>Fresh fruit cross-sections were transferred to each substrate by direct contact for 10 seconds before analysis using a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=5&amp;search=DESI+XS"><strong>Waters DESI XS ion source</strong></a> coupled to a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/2433"><strong>Waters Xevo™ MRT P10 high-resolution mass spectrometer</strong> </a>operating in negative electrospray ionization mode. MS imaging was performed with a <strong>50 µm pixel size</strong>, using a <strong>95:5 methanol/water</strong> spray solvent, while sucrose (m/z 341.1089) served as the lock mass for accurate mass calibration. High mass resolution enabled tentative assignment of metabolites such as citric acid, asparagine, malic acid, glutamic acid, palmitic acid, stearic acid, and sucrose.</p><p>The comparison of transfer substrates revealed significant differences in metabolite localization and image quality. Glass slides produced the strongest metabolite signals but also exhibited considerable metabolite delocalization during sample transfer, reducing spatial accuracy. Nitrocellulose membranes transferred a broader range of metabolites and provided complementary chemical information, although they also showed greater analyte spreading and suffered physical damage after a single DESI acquisition. In contrast, <strong>Poropare plates</strong> minimized metabolite delocalization and preserved fine structural details within the fruit cross-sections, despite producing somewhat lower signal intensities than glass slides.</p><p>The study concludes that direct transfer combined with DESI MSI is an effective approach for imaging soft fruit tissues that are otherwise difficult to analyze because of their high water content. Among the evaluated substrates, <strong>Hamamatsu Poropare plates</strong> provided the best balance between spatial resolution and preservation of metabolite localization, making them particularly suitable for high-resolution metabolomic imaging, while high mass accuracy facilitated confident tentative metabolite identification.</p></div></div></div></div></div></div></section></div></div>

News from LabRulezLCMS Library - Week 30, 2026

<p><strong>Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 10th August 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, Thermo Fisher Scientific / ASMS </strong>and<strong> Waters Corporation / ASMS </strong>and application notes by<strong> Metrohm and Shimadzu!</strong></p><h3><strong>1. Agilent Technologies / ASMS: Impurity Monitoring of GLP-1 Therapeutic Peptide RetatrutideUsing a 2D-LC/TOF System</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/glp_1_impurity_monitoring_2d_lc_tof_asms_2026_mp_388_agilent_eadce3f244.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Retatrutide is a next-generation triple agonist (GLP1/GIP/glucagon) and an emerging candidate among multivalent GLP1–based therapeutics. Its structural complexity and diverse impurity profile challenge conventional one-dimensional LC methods, which often lack sufficient resolving power to separate closely related impurities, sequence variants, and synthetic byproducts.&nbsp;</p><p>In this study, we demonstrate an Agilent multiple heart-cutting 2D-LC platform that enables rapid online solvent exchange and enhanced chromatographic selectivity, allowing targeted isolation of impurity peaks from a TFA-based first-dimension separation. When coupled with the Agilent 6230C Accurate Mass TOF LC/MS system, the workflow delivers fast, sensitive, and accurate mass profiling under MS-friendly second-dimension conditions, providing a robust solution for comprehensive impurity characterization of GLP-1 peptides such as Retatrutide.</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?instrumentationItems=39&amp;mainauthorItems=10&amp;search=1290+infinity+III+2D-LC"><strong>Agilent 1290 Infinity III 2D-LC system</strong></a><strong> comprised the following modules:&nbsp;</strong><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></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><h4><strong>Conclusions</strong></h4><p>Agilent 2D-LC with multiple heart-cutting effectively bridges TFA-driven chromatographic performance with FA-compatible MS detection, overcoming ion suppression limitations.&nbsp;</p><p>The integrated 2D-LC/Accurate Mass TOF workflow delivers confident impurity identification and robust characterization, supporting peptide analysis in both QA/QC and development environments.</p><h3><strong>2. Metrohm: Vinylene carbonate water determination via coulometric Karl Fischer titration</strong></h3><p><strong>Water determination in vinylene carbonate by Karl Fischer coulometric titration made simple with alcohol-free reagents</strong></p><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/an_k_076_download_8658665940.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Vinylene carbonate (vc) is commonly used as a battery electrolyte additive in lithium-ion batteries. Its popularity is due to the positive impact it has on anode cyclability and reduction of irreversible capacity. Small amounts of moisture in Li-ion batteries can significantly decrease performance and lifetime. Therefore, moisture levels in electrolyte solutions and their additives are controlled for use in manufacturing. Until recent years, accurate moisture quantification in vinylene carbonate samples was difficult due to side reactions. With the commercial development of alcohol-free coulometric Karl Fischer reagents, accurate determination is now much more accessible to end-users.</p><h4><strong>EXPERIMENTAL</strong></h4><p>This analysis was performed using an <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=220&amp;search=852+titrando"><strong>852 Titrando</strong></a> (2.852.0060) instrument, equipped with the electrodes listed in Table 1.</p><h4><strong>CONCLUSION&nbsp;</strong></h4><p>During battery manufacturing, control of all moisture present is important to guarantee adequate performance in the finished product. More easily quantifying the moisture contribution from additives such as vinylene carbonate makes this a much simpler task. The introduction of alcohol-free Karl Fischer reagents accomplishes this effectively, allowing the generation of accurate and repeatable moisture data—even for a difficult sample matrix.</p><h3><strong>3. Shimadzu: Non-Targeted Analysis of Metabolites in Alcoholic Beverages Using LabSolutions Insight Profiler</strong></h3><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/an_01_01241_en_57b43edb57.pdf"><strong>Full PDF for download</strong></a></li></ul><h5><strong>User Benefits</strong></h5><ul><li>Using an LC-QTOFMS system with LabSolutions Insight Profiler non-targeted analysis software enables comprehensive comparative analysis of metabolites in food samples.&nbsp;</li><li>Principal component analysis and volcano plots can be used to detect characteristic features in each sample.&nbsp;</li><li>A variety of functions, including MS/MS spectral library searching, enable easy qualitative analysis of components.</li></ul><p>Metabolomics, a technique that analyzes metabolites in living organisms, is increasingly being applied in the food industry for product development and quality control. A particularly powerful approach is non-targeted metabolomics, which aims to analyze not only known metabolites but also unknown compounds without prior assumptions to reveal unexpected chemical differences between samples. Quadrupole time-of-flight mass spectrometry (QTOFMS), with its accurate-mass capability, is well suited for this purpose. However, the large volume of generated data creates challenges for feature identification and data comparison.&nbsp;</p><p>This article presents a case study in which LabSolutions Insight Profiler, a software platform for non-targeted analysis, was used to characterize compounds in beer and beer-like beverages. Peak detection, retention-time alignment, and library searching were performed across multiple datasets. Multivariate analysisrevealed compound profiles that likely reflect the differences in raw materials and manufacturing processes, demonstrating the utility of the approach for beverage characterization.</p><h4><strong>LabSolutions Insight Profiler&nbsp;</strong></h4><p><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=LabSolutions+Insight+Profiler"><strong>LabSolutions Insight Profiler </strong></a>is software for batch processing of LC-QTOFMS data that enables non-targeted analysis through an intuitive workflow. It readily supports easy spectral feature detection, alignment, statistical processing, filtering, list screening, MS/MS spectra-based library searching, principal component analysis (PCA), displaying volcano or box plots, and more. Fig. 1 shows the data analysis workflow.</p><h4><strong>Analysis Conditions&nbsp;</strong></h4><p><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=nexera+X3"><strong>Nexera X3 UHPLC</strong></a> and<strong> </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=9030"><strong>LCMS-9030 systems</strong></a> were used as the analytical instruments. The LC method included in the LC/MS/MS Method Package for Primary Metabolites was used as the method. The data-dependent acquisition (DDA) mode was used to simultaneously acquire precursor m/z and MS/MS data. Table 2 shows the analysis conditions.</p><h4><strong>Conclusion&nbsp;</strong></h4><p>In this study, compounds in various beer and beer-like beverages were analyzed by non-targeted analysis using an LCMS-9030 quadrupole time-of-flight mass spectrometer system. Analyzing compounds characteristic of each sample with LabSolutions Insight Profiler software confirmed that the resulting profiles reflected differences in raw materials and manufacturing methods.&nbsp;</p><p>LabSolutions Insight Profiler enables efficient non-targeted analysis by processing LC-QTOFMS data with an intuitive workflow. This workflow allows for objective evaluation of the taste, quality, and nutritional value of food products. Furthermore, this method is applicable not only to the analysis of compounds in beverages but also to non-targeted analysis of other types of samples.</p><h3><strong>4. Thermo Fisher Scientific / ASMS: High throughput targeted proteomic workflows with Adaptive Retention Time on the </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1959"><strong>Orbitrap Astral Zoom MS</strong></a></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_004699_lsms_art_astral_zoom_proteomics_po004699_asms2026_na_en_78767efdc9.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Advances in MS instrumentation have accelerated the identification of biomarkers, but validating and translating these discoveries into assays for clinical use remains challenging and typically requires robust and accurate targeted approaches. While these often rely on predetermined reference target retention time (RT) windows, analytical column degradation or replacement, aging solvents, sample matrix effects, etc. can cause peptide RT shifts between injections and across longer-term analyses. To accommodate this, targeted MS experiments generally schedule wide target RT windows (&gt;1 min) or rely on heavy labeled peptides to trigger acquisition of endogenous peptides. Both greatly limit the number of quantifiable targets per assay. To overcome this, we utilize adaptive RT (aRT) alignment for live correction of RTs together with the speed and sensitivity of the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1959"><strong>Orbitrap Astral Zoom MS</strong></a>. This allows reproducible tracking of &gt;2000 targets with narrow RT windows, high throughputs, such as 300 samples per day (SPD), and hybrid workflows combining targeting, of up to 1000 targets, and discovery.</p><h4><strong>Materials and methods&nbsp;</strong></h4><h5><strong>LC-MS Experiments&nbsp;</strong></h5><p>Experiments were performed using Thermo Scientific Pierce HeLa protein digest resuspended in 0.1% TFA, or individual or pooled human plasma samples. Peptides were separated using a 15 cm Thermo Scientific PepMap analytical columns (150 µm inner diameter) and the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/398"><strong>Thermo Scientific Vanquish Neo UHPLC system</strong></a> at throughputs of 60 or 300 SPD. The liquid chromatography (LC) system was interfaced with the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1959"><strong>Orbitrap Astral Zoom MS.</strong></a> Reference runs using MS1 and/or data-independent-acquisition (DIA) scans were acquired to obtain reference aRT spectra, and to determine target RTs. For aRT alignment, methods consisting of aRT MS1 and/or DIA scans, as well as targeted MS2 scans with RT correction were optimized. For hybrid DIA workflows, we used aRT MS1 scans for reference acquisition and alignment, combined with narrow window DIA and a targeted MS2 experiment. Observed RT shifts were caused by natural column degradation, sample matrix effects or when indicated induced by variation in gradient flow rate and buffer composition.&nbsp;</p><h5><strong>Data analysis&nbsp;</strong></h5><p>Reference DIA data was analyzed using Spectronaut. Targets lists were then generated from the search output by selecting varying number of targets evenly distributed across the RT dimension. Target RTs for smaller scale assays were manually validated and exported from Skyline, while target RTs for large scale assays were exported from Spectronaut. For alignment runs, DIA and PRM data were analyzed using Spectronaut and Skyline respectively.</p><h4><strong>Conclusions&nbsp;</strong></h4><ul><li>Adaptive RT alignment on Orbitrap Astral Zoom MS shows robustness against various sources of RT shifts and has high target recovery even for thousands of targets&nbsp;</li><li>aRT enables the use of target acquisition windows &lt;15 s for robust targeting at high throughput&nbsp;</li><li>aRT Hybrid DIA allows targeting of up to 1000 targets with minimal cost to DIA depth</li></ul><h3><strong>5. Waters Corporation / ASMS: BOOSTING SENSITIVITY IN TOP-DOWN PROTEIN STUDIES USING CYCLIC IMS-ENABLED WIDEBAND ENHANCEMENT</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/720009471_51f2e8b546.pdf"><strong>Full PDF for download</strong></a></li></ul><div><div><section><div><div><div><div><div><div><p>The poster evaluates <strong>Wideband Enhancement (WBE)</strong> as a way to improve sensitivity in top-down protein mass spectrometry. In top-down workflows, intact proteins are fragmented directly, but the signal is distributed across multiple charge states, isotope envelopes, and numerous product ions, which can limit MS/MS sensitivity. WBE addresses this problem by synchronizing ion packets leaving the ion mobility region with the time-of-flight pusher, thereby increasing the number of ions that reach the detector within a selected <i>m/z</i> range.</p><p>Experiments were performed using a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/2434"><strong>Waters Cyclic IMS P20 Mass Spectrometer</strong></a> equipped with an <strong>electron capture dissociation cell</strong>. Samples were introduced by direct infusion in positive electrospray ionization mode. Two proteins were studied under denaturing conditions: bovine ubiquitin, approximately 8.5 kDa, and bovine carbonic anhydrase II, approximately 29 kDa. Single-pass and bypass ion mobility measurements were carried out with and without WBE, using both <strong>collision-induced dissociation (CID)</strong> and <strong>electron capture dissociation (ECD)</strong>. Data were acquired in <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/635"><strong>MassLynx</strong></a><strong> v4.2</strong> and processed using <strong>DriftScope</strong> and the <strong>waters_connect UNIFI peptide-mapping workflow</strong>.</p><p>For ubiquitin, WBE substantially increased the intensity of CID- and ECD-derived fragment ions. Improvements of up to tenfold were observed across the c- and z-ion series, while bypass ion mobility mode produced the largest overall sensitivity gain. The results indicate that WBE can enhance fragment detection while retaining the benefits of cyclic ion mobility separation and without sacrificing mass-spectral resolution.</p><p>The larger carbonic anhydrase II protein also showed marked improvements. CID fragment signals increased by up to tenfold for lower-<i>m/z</i> ions and about eightfold for higher-<i>m/z</i> fragments. Because ECD is generally less efficient than CID, the benefit of WBE was especially evident in ECD experiments: sequence coverage increased from <strong>54% without WBE to 70% with WBE</strong> in bypass mode. The authors conclude that combining WBE with complementary CID and ECD fragmentation on the Cyclic IMS P20 enables more sensitive and comprehensive top-down protein characterization.</p></div></div></div></div></div></div></section></div></div>

News from LabRulezLCMS Library - Week 33, 2026

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A Hands‑On Spectroscopy Workshop in Agilent Wood Dale Center of Excellence

PFAS Seminar Series

Shimadzu at 2026 AOAC Annual Meeting & Exposition

ICP-MS for Hybrid Photoelectrode Characterization

Addressing LC Method Optimization - Scaling For Efficiency, Throughput And Reproducibility

Proteoform Studio software, top down made simple