Impact of mobile and stationary phases on siRNA duplex stability in liquid chromatography

<p><strong>Welcome to the Column Coach, a tool to help you with reversed-phase column selectivity. This tool uses the Neue Test to help compare column hydrophobicity and selectivity, based on a set of standard conditions using the same test mixtures.</strong></p><p><strong>For more details about the Neue Test, please refer to:</strong></p><ul><li><a target="_blank" rel="noopener noreferrer" href="http://onlinelibrary.wiley.com/doi/10.1002/jssc.200390025/full">Neue, U. D.; Van Tran, K.; Iraneta, P. C.; Alden, B. A. Characterization of HPLC Packings. J. Sep. Sci. 2003, 26, 174-186.</a></li><li><a target="_blank" rel="noopener noreferrer" href="https://www.sciencedirect.com/science/article/pii/S0021967399004343">Neue, U. D.; Alden, B. A.; Walter, T. H. Universal Procedure for the Assessment of the Reproducibility and the Classification of Silica-based Reversed-phase Packings II. Classification of Reversed-phase Packings. J. Chromatogr. A. 1999, 849, 101-116.</a></li></ul><h4><strong>How to read the graph?</strong></h4><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_Corporation_Column_Coach_76473457ee.jpg" alt="Waters Corporation: Column Coach" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_Corporation_Column_Coach_76473457ee.jpg 184w," sizes="100vw" width="1000" height="850"></p><p>All trademarks are the property of their respective owners. Specifications were obtained from manufacturer's public literature. Waters does not endorse or represent the accuracy or reliability of any manufacturers' information, and presentation of this information does not imply any relationship between Waters and any manufacturer. Your use of this information is at your sole risk. This tool and its contents are provided and distributed on an 'as is' basis without warranties of any kind either express or implied, including without limitation, implied warranties or merchantability of fitness for a particular purpose.</p><h4><strong>You can search for the right column for your application by:</strong></h4><ul><li><strong>Compound Class</strong> - Want to find a column for a specific class of compounds? Select a compound class from the drop down. Next select a specific sub-compound class and the recommended Waters columns for that given sub-compound class will be provided.</li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_Corporation_Column_Coach_Search_by_Compound_Class_ac76b423b9.jpg" alt="Waters Corporation: Column Coach - Search by Compound Class" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_Corporation_Column_Coach_Search_by_Compound_Class_ac76b423b9.jpg 245w," sizes="100vw" width="1458" height="666"></p><ul><li><strong>USP Designation</strong> - Our quality mission is to ensure that the Waters' columns you use today are the most reproducible and reliable LC columns available. In this section, you will find all Waters columns that are registered according to classifications prescribed in the United States Pharmacopeia (USP).</li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_Corporation_Column_Coach_Search_by_USP_Designation_1a8588c591.jpg" alt="Waters Corporation: Column Coach - Search by USP Designation" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_Corporation_Column_Coach_Search_by_USP_Designation_1a8588c591.jpg 190w," sizes="100vw" width="1464" height="1207"></p><ul><li><strong>Manufacturer of your Existing Column</strong></li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_Corporation_Column_Coach_Search_by_Manufacturer_of_your_Existing_Column_cebd6ae8c0.jpg" alt="Waters Corporation: Column Coach - Search by Manufacturer of your Existing Column" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_Corporation_Column_Coach_Search_by_Manufacturer_of_your_Existing_Column_cebd6ae8c0.jpg 210w," sizes="100vw" width="1446" height="1073"></p><ul><li><strong>Selectivity Chart</strong></li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_Corporation_Column_Coach_Search_by_Selectivity_Chart_5883ba13ab.jpg" alt="Waters Corporation: Column Coach - Search by Selectivity Chart" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_Corporation_Column_Coach_Search_by_Selectivity_Chart_5883ba13ab.jpg 245w," sizes="100vw" width="1478" height="904"></p>

Waters HPLC Column Coach

Development and thorough evaluation of a multi-omics sample preparation workflow for comprehensive LC-MS/MS-based metabolomics, lipidomics and proteomics datasets

<p><strong>Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 11th August 2025? 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 </strong>and <strong>Thermo Fisher Scientific / ASMS </strong>and application notes by <strong>Shimadzu </strong>and<strong> Waters Corporation!</strong></p><h3><strong>1. Agilent Technologies / ASMS: Fully Automated Sensitive Quantitation of PFAS in Seafood for Regulatory Screening Using Triple Quadrupole LC/MS&nbsp;</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/ps_6495d_pal3_pfas_asms_2025_mp_240_en_agilent_234d300c8d.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Per- and polyfluoroalkyl substances (PFAS) are extensively found in marine life, posing health risks through seafood consumption. Seafood is one of the regulated matrices by the U.S. FDA, EU, EURL POPs, and AOAC. Regulatory and standard-making bodies like the U.S. FDA, EFSA, EURL POPs, and AOAC have set stringent guidelines for PFAS levels in seafood. Detecting trace PFAS level is challenging due to complex sample preparation like QuEChERS extraction followed by SPE cleanup, and evaporation/reconstitution. The manual steps can be labor-intensive and error-prone, affecting accuracy and reliability. This study developed a fully automated workflow for PFAS quantitation in seafood – shrimp.</p><h4><strong>Experimental</strong></h4><h5><strong>Instrumentation&nbsp;</strong></h5><p>An integrated 160 cm <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=+PAL3"><strong>PAL3 Series 2 RTC autosampler</strong></a> coupled with a<a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=+6495D+LC%2FMS"><strong> 6495D LC/TQ</strong></a> (Figure 1) was employed. The PAL3 platform performed automated sample preparation, while TQ data acquisition and analysis were conducted in parallel. LC system comprised of two modules; an Agilent 1290 Infinity II high-speed pump and an Agilent 1290 Infinity II multicolumn thermostat. The study utilized the following tools and modules: two PAL park stations with three liquid syringe tools, a dilutor tool, a micro-SPE tool, an LC/MS tool, a vortex mixer, a centrifuge, a dilutor multi, a tray cooler (for 2/10/20 mL vials), tray holders with rack R60 (for 10/20 mL vials), a micro-SPE tray (for 2 mL vials and micro-SPE cartridges), a solvent module, a fast wash module, and an LC injection valve. The details of various consumables used for this study are provided elsewhere.1</p><h4><strong>Conclusions</strong></h4><ul><li>The integrated system allows sample preparation and data analysis to run in parallel, offering a streamlined workflow and improving productivity for routine laboratory operations.&nbsp;</li><li>The automated workflow significantly reduces manual intervention, minimizing human error and enhancing the precision of the analysis.&nbsp;</li><li>The integration of automated sample preparation techniques with the highly sensitive 6495D LC/TQ ensures consistent and reproducible results, which are critical for regulatory compliance.&nbsp;</li><li>The workflow demonstrated excellent analytical performance meeting the stringent regulatory requirements and recommendations for PFAS in seafood matrices set by the U.S. FDA, EU, EURL POPs, and AOAC.</li></ul><h3><strong>2. Shimadzu: Determination of PFASs in Food Contact Material by </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=LCMS-8050"><strong>LCMS-8050</strong></a></h3><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/an_03_lcmsms_961_en_34db9cd4cb.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Due to the unique physical and chemical properties of per- and polyfluoroalkyl substances (PFASs), such as reducing surface tension, good stability, hydrophobicity, and hydrophilic, so it is very widely used in the field of food contact materials. It is mainly used for the production of plastic film, PVC food packaging, waterproof, and oil-proof coating on the surface of food packaging (such as waterproof oil paper products) and non-stick cooker coating, etc. Research has shown that PFASs will migrate from food contact materials to food, and then contaminate the food that comes into contact with it, entering the human body. PFASs are very stable, difficult to be metabolized and biodegraded in the body, and exhibit the characteristics of persistence, accumulation, and long-distance migration. It can be accumulated and amplified in the food chain and enriched in the organisms, endangering human health.&nbsp;</p><p>Refer to the sample preparation method in GB 31604.35-2016, determination of both Perfluorooctane Sulfonates (PFOS) and Perfluorooctanoic Acid (PFOA), an analytical method for the determination of 17 perfluorinated compounds in food contact materials was built.</p><h4><strong>Analysis Conditions</strong></h4><h5><strong>System: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=Nexera+XS"><strong>Nexera XS&nbsp;</strong></a></h5><ul><li>Column : Shim-pack GIST C18-AQ HP (2.1 mm I.D.×100 mm, 1.9 μm)</li><li>Temperature : 30 ˚C&nbsp;</li><li>Injection volume : 5 µL&nbsp;</li><li>Mobile phases : A-5 mM ammonium acetate in Water B-ACN&nbsp;</li><li>Flow rate : 0.3 mL/min&nbsp;</li><li>Elution mode : Gradient elution&nbsp;</li><li>Time program (%B) : 15% (0 min) → 15% (2.5 min) →98% (10-12 min) → 15% (12.1-15 min)&nbsp;</li></ul><h5><strong>System: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=LCMS-8050"><strong>LCMS-8050</strong></a><strong> (ESI Negative)&nbsp;</strong></h5><ul><li>Nebulizing gas : 3 L/min&nbsp;</li><li>Drying gas : 10 L/min&nbsp;</li><li>Heating gas : 10 L/min&nbsp;</li><li>DL temp. : 250 ˚C&nbsp;</li><li>Heat block temp. : 400 ˚C&nbsp;</li><li>Interface temp. : 300 ˚C</li></ul><h4><strong>Conclusion&nbsp;</strong></h4><p>Utilizing the LCMS-8050 system for quantitative analysis of PFASs in Food Contact Material revealed that the method can accurately determine PFASs levels within a broad concentration range of 0.05 to 5 ng/mL. This approach exhibits several notable advantages, including excellent repeatability, stability, and reliability.</p><h3><strong>3. Thermo Fisher Scientific / ASMS: Leveraging advanced mass spectrometry technology with </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1959"><strong>Orbitrap Astral Zoom MS</strong></a><strong> for in depth immunopeptidome profiling&nbsp;</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_414_advanced_mass_spec_technology_orbitrap_astral_zoom_po414_asms2025_en_3ca6419434.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Immunopeptidomics is the study of the peptides presented by MHC molecules on the surface of cells. These MHC peptides have major implications for many areas of research, including immunotherapy and personalized medicine. Mass spectrometry allows for direct immunopeptidomics analysis, enabling simultaneous identification and quantification of thousands of MHC peptides in a single run. The new <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1959"><strong>Orbitrap Astral Zoom MS </strong></a>has enabled new levels of sensitivity and selectivity to provide deeper insights into the immunopeptidome.</p><h4><strong>Materials and methods</strong></h4><h5><strong>LC-MS/MS method&nbsp;</strong></h5><p>Peptides were separated on a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/398"><strong>Vanquish Neo UHPLC System</strong></a> using Aurora Ultimate column (25 cmx 75µm). Total run time was 72 min. Thermo Scientific EASY-Spray Ion Source was used coupled to the Thermo Scientific FAIMS Pro Duo interface. Peptides were analyzed by the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1959"><strong>Orbitrap Astral Zoom MS</strong></a>.</p><h5><strong>Data Processing&nbsp;</strong></h5><p>The data analysis was performed using PEAKS Studio 12 with DeepNovo Peptidome workflow for database search and de novo peptides identification. Spectra were searched against UniProt human database (20,607 sequences) with no-enzyme option. The sequence motif and binding properties of 9-mer peptides were analyzed using MHCMotifDecon 1.2 and NetMHCpan 4.0.</p><h4><strong>Conclusions&nbsp;</strong></h4><ul><li>Orbitrap Astral Zoom MS equipped with FAIMS Pro Duo interface and coupled to Vanquish Neo UHPLC system provides high sensitivity and the dynamic range necessary for deeper coverage of immunopeptidomic samples</li><li>The low input application mode on Orbitrap Astral Zoom MS offers the sensitivity necessary to analyze low levels of material equivalent to samples ectracted from tissue biopsy samples</li><li>Loading capacity is not limited by MS allowing analyses from ultra low (1e5) to high (1e8) loads of sample&nbsp;</li><li>FAIMS enables the analysis of singly and multiply charged ions enhancing the immunopeptidome coverage&nbsp;</li><li>AGC modulates injection times for each peptide, producing high quality spectra across a wide range of loading levels. This spectral quality is crucial for downstream analysis and confident identification and quantitation of neoantigens</li></ul><h3><strong>4. Waters Corporation: Optimization of Detector Parameters to Improve Sensitivity using the </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/717"><strong>Alliance™ iS HPLC System </strong></a><strong>with PDA Detector</strong></h3><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/720008901_en_70df6fce93.pdf"><strong>Full PDF for download</strong></a></li></ul><h4><strong>Benefits&nbsp;</strong></h4><ul><li>The Alliance iS HPLC System with PDA Detector allows the user to easily modify PDA settings to obtain optimal performance, including signal-to-noise</li><li>The Alliance iS HPLC System with PDA Detector has a range of parameters, including variable slit width and wide resolution range for method optimization</li><li>Optimization of the PDA settings produced a S/N ratio increase of 7x</li></ul><p>Default PDA instrument method settings are frequently used for running HPLC methods. When running a method for the first time, these default settings are a good starting point and often produce suitable chromatography. However, detector parameters may need to be modified to obtain optimal separations. In this study, the USP method for organic impurities in ibuprofen tablets was used to show how the Alliance iS HPLC System with PDA Detector parameters may be optimized to improve sensitivity. The USP S/N ratio was used to define sensitivity. The method is a reversed-phase isocratic method with UV detection (254 nm). The method’s system suitability criteria includes a S/N ratio of not less than (NLT) 10 for a 5-ppm solution of ibuprofen.&nbsp;</p><p>The Alliance iS HPLC System with PDA Detector settings that were evaluated and optimized for their impact on sensitivity included data rate, filter time constant, slit width, resolution, and absorbance compensation.&nbsp;</p><ul><li><strong>Data rate:</strong> The data rate defines the rate that the detector collects data, measured in hertz (Hz). The data rate should be set to yield reproducible peak area and retention times. Data rates that are too low will result in poorly defined peaks; high data rates may result in increased noise. Generally, the data rate is set to collect 25-50 points across the narrowest peak in the chromatogram. The data rate of the Alliance iS HPLC Systém with PDA Detector has settable values of 1–160 Hz. The default data rate is 10 Hz.&nbsp;</li><li><strong>Filter time constant: </strong>The filter time constant is a noise filter that filters out high frequency noise. Faster time constants produce narrower peaks and remove less baseline noise. Slower time constants result in broader peaks and decreased baseline noise. The Alliance iS HPLC System with PDA Detector has the following filter time constant options: no filter, slow, normal, fast, and custom. The default selection is normal.</li><li><strong>Slit width:</strong> Slit width is an added feature available in the Alliance iS HPLC System with PDA Detector. The slit width determines the amount of light that reaches the photodiode array sensor which impacts resolution. Smaller slit widths result in improved resolution. Larger slit widths produce less noise and increased sensitivity, but at the cost of lower resolution. The Alliance iS HPLC System with PDA Detector has a variable slit, with available slit width sizes of 35 µm, 50 µm, 100 µm, and 150 µm. The detector’s default slit width is 50 µm.</li><li><strong>Resolution:</strong> The resolution setting determines bandwidth, which is the number of diode responses that are averaged when calculating the absorbance at a specified wavelength. Larger values result in less noise and a better S/N ratio, but they also produce decreased spectral resolution and can impact linearity. Resolution on the Alliance iS HPLC System with PDA Detector is settable over a range of 1<span>–</span>20 nm. The default value is 4 nm.&nbsp;</li><li><span style="font-family:&quot;Arial&quot;,sans-serif;"><strong>A</strong></span><strong>bsorbance compensation</strong>: The absorbance compensation feature of the Alliance iS HPLC System with PDA Detector provides a way to reduce non-wavelength dependent noise. This is achieved by collecting absorbance data over a user-specified wavelength range where there is little or no absorption and calculating the average absorbance. This average absorbance is then subtracted from the absorbance value. The Alliance iS HPLC System with PDA Detector default setting for absorbance compensation is ‘off’.</li></ul><p>Using the USP method, each of these detector settings were individually evaluated and optimized. A method was created using the optimized settings and the results were compared to those obtained using default values.</p><h4><strong>Experimental</strong></h4><h5><strong>LC Conditions&nbsp;</strong></h5><ul><li><strong>LC system: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/717"><strong>Alliance iS HPLC System</strong></a> with PDA Detector&nbsp;</li><li><strong>Detection:</strong> PDA Detector with 10 mm flow cell</li><li><strong>Column: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1823"><strong>XBridge™ BEH™ C18</strong></a>, 250 x 4.6 mm; 5 µm (p/n: 186003117)</li></ul><h5><strong>Data Management&nbsp;</strong></h5><ul><li><strong>Chromatography data system:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/627"><strong>Empower™ Chromatography Data System (CDS)</strong></a></li></ul><h4><strong>Conclusion</strong></h4><p>While default PDA instrument method settings may be suitable for routine HPLC analysis, they may need to be adjusted to produce optimal separations. This study demonstrated how the Alliance iS HPLC System with PDA Detector allows the user to easily modify PDA settings to obtain optimal separations. Using the USP method for organic impurities in ibuprofen tablets, the data rate, filter time constant, slit width, resolution, and absorbance compensation settings were optimized. The optimized settings produced a 7-fold increase in sensitivity over that obtained using default values.</p>

News from LabRulezLCMS Library - Week 33, 2025

<p><strong>Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 15th September 2025? 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 application notes by <strong>Agilent Technologies, Shimadzu </strong>and <strong>Waters Corporation </strong>and<strong> </strong>poster by <strong>Thermo Fisher Scientific / HPLC Symposium!</strong></p><h3><strong>1. Agilent Technologies: A Direct Aqueous Injection Method for Contaminants in Drinking and Nonpotable Water</strong></h3><p><strong>Analysis of acrylamide, haloacetic acids, and β-estradiol in water with an Agilent 6495D LC/TQ</strong></p><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/an_aquesous_injection_method_water_6495d_lc_tq_5994_7973en_agilent_1_24e3a63a15.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Recent regulations, such as EU 2020/218, have included the addition of new emerging contaminants in drinking water and raw waters, which supply water-treatment works. Some of these new contaminants to be analyzed include acrylamide, HAAs, and β-estradiol. However, acrylamide and HAAs can be difficult to analyze due to their high polarity and high interferences from hard water samples. Additionally, β-estradiol requires a very low level of detection (1 ng/L) compared to the other compounds (acrylamide 0.1 µg/L and HAAs 60&nbsp;µg/L). The analysis uses a&nbsp;1290 Infinity II LC coupled to a&nbsp;6495D LC/TQ for analysis of water samples.</p><h4><strong>Experimental</strong></h4><h5><strong>Instrumentation&nbsp;</strong></h5><p>Chromatographic separation was performed using an <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=InfinityLab+Poroshell+120+Aq-C18"><strong>Agilent&nbsp;InfinityLab Poroshell 120 Aq-C18</strong></a>, 3.0 × 150&nbsp;mm, 2.7&nbsp;µm (part&nbsp;number 693675-742) installed on the&nbsp;1290&nbsp;Infinity II LC system. The individual modules of the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=1290+Infinity+II+LC"><strong>1290 Infinity II LC system</strong></a>&nbsp;included:</p><ul><li>Agilent 1290 Infinity II high‑speed pump (G7120A)&nbsp;</li><li>Agilent 1290 Infinity II autosampler (G7167B)&nbsp;</li><li>Agilent 1290 Infinity II multicolumn thermostat (G7116B)&nbsp;</li></ul><p>The <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=6495+LC%2FTQ"><strong>6495 LC/TQ</strong></a> with an Agilent&nbsp;Jet Stream (AJS) electrospray ion source was operated in dynamic multiple reaction monitoring (dMRM) mode. This mode allows more MRM transitions if future development is required for additional compounds. The LC/TQ autotune was performed in both unit and wide modes. All data acquisition and processing were performed using Agilent&nbsp;MassHunter software (version&nbsp;12).</p><h4><strong>Conclusion&nbsp;</strong></h4><p>This application note describes a highly sensitive and reproducible method for the fast and reliable quantitation of the listed compounds in water by direct injection. The dMRM method was created and optimized using Agilent MassHunter software and allows the addition of more MRM transitions if future development is required for additional compounds. An Agilent 1290 Infinity II LC coupled to an Agilent 6495D LC/TQ was used for the analysis. The 12-minute LC gradient method using an Agilent Aq-C18 column offered good chromatographic separation and RT distribution of all targets. The LC/TQ data acquisition was in dMRM mode with fast polarity switching for the most efficient use of instrument cycle time. The method performance was verified based on requirements for calibration curve linearity, instrument LOD, recovery, and precision. The results demonstrate the ability of the quantitative analytical method for emerging contaminants in water by direct injection.</p><h3><strong>2. Shimadzu: Short-Chain Oligonucleotide Analysis Using Supercritical Fluid Chromatography</strong></h3><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/ap_aplnote100_en_23b7bffc7e.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Oligonucleotide therapeutics are drugs in which oligonucleotide is the active ingredient. Oligonucleotide therapeutics are manufactured through chemical synthesis, but impurities are produced during the process, so measurement and characterization of the impurities are required. Currently, reversed-phase ion-pair chromatography is often used for this analysis, but it is difficult to separate some impurities that are structurally similar to the target product. Therefore, a separation method with selectivity different from existing methods is required.&nbsp;</p><p>SFC is an analytical method that employs carbon dioxide as the mobile phase at temperatures and pressures above the respective critical points. Compared to liquids, carbon dioxide is reported to have high resolution and special selectivity as a component of SFC mobile phase because of its low viscosity and high diffusivity.&nbsp;</p><p>On the other hand, since the mobile phase containing carbon dioxide has low polarity, SFC was considered unsuitable for the analysis of highly polar compounds. However, in recent years, the composition of the modifier mixed with carbon dioxide and the choice of stationary phase have made it possible to analyze highly polar compounds,such as peptides and nucleotides2).&nbsp;</p><p>In this study, we evaluated the applicability of SFC to oligonucleotide analysis. Single-strand oligonucleotides of around 20-mer are often used as oligonucleotide therapeutics, but due to the extremely high polarity of the molecules and the difficulty of analysis, we first used 4-mer oligonucleotides for evaluation and investigated their basic retention behaviors.</p><h4><strong>Experimental</strong></h4><ul><li><strong>System:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=LCMS-9030"><strong>LCMS-9030</strong></a></li></ul><h4><strong>Conclusions&nbsp;</strong></h4><p>To evaluate the applicability of SFC for oligonucleotide analysis, we developed a separation method using 4-mer oligonucleotides with varying PS contents as model compounds and investigated their basic retention characteristics. First, we identified conditions that allowed oligonucleotides to elute, finding that a methanol-water mixture containing 50 mmol/L ammonium formate enabled elution and peak detection. Using these conditions, we analyzed T4 with varying PS contents on 12 columns, finding good separation with the Diol II column. To apply this method to more polar sequences, we re-evaluated modifier conditions and found that adding aminoethanol improved peak shape. Applying the optimized method to 14 oligonucleotide sequences showed that sequences with two or fewer G or C bases were suitable for this method. Additionally, higher-polarity sequences showed stronger retention, while oligonucleotides with varying PS contents showed weaker retention with higher polarity, indicating unique retention behavior for PS modifications. The high selectivity for sequences with different PS contents suggests that SFC can be applied to separating impurities with a PS linkage replaced by a PO linkage that can be included in PS-modified oligonucleotides in the synthesis process.&nbsp;</p><p>In the future study, the applicability of SFC to longer oligonucleotidesthan 4-mer will be evaluated.</p><h3><strong>3. Thermo Fisher Scientific / HPLC Symposium: Sensitive LC-MS method for the quantitative analysis of semaglutide and liraglutide in human plasma</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_sensitive_lc_ms_method_quantitative_analysis_semaglutide_po_en_592e7dcc4a.pdf"><strong>Full PDF for download</strong></a></li></ul><p>GLP-1 analogs have emerged as pivotal therapeutic agents in the management of type 2 diabetes and obesity. These analogs mimic the physiological actions of endogenous GLP-1, including enhancing glucose-dependent insulin secretion, suppressing glucagon release, delaying gastric emptying, and promoting satiety. Given their significant clinical utilities, accurate detection and quantification of GLP-1 analogs in human plasma are essential for pharmacokinetic studies, therapeutic monitoring, and ensuring patient safety. This study aims to develop a sensitive LC-MS method for the detection and quantitation of semaglutide and liraglutide in human plasma.</p><h4><strong>Materials and methods</strong></h4><h5><strong>Mass Spectrometry</strong></h5><p><span>&nbsp;</span>See Table 2 for detailed MS source and scan settings for the analysis GLP-1 analogs on <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/404"><strong>TSQ Altis Plus MS</strong></a>.&nbsp;</p><h5><strong>Data Analysis&nbsp;</strong></h5><p><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/74"><strong>Thermo Scientific Enterprise </strong></a>compliance ready Chromeleon CDS 7.3.2 software was used for all instrument control, data acquisition, processing, and reporting.</p><h4><strong>Conclusions&nbsp;</strong></h4><p>We demonstrated a comprehensive LCMS solution for highly sensitive quantitation of GLP-1 analogs in human plasma using the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/744"><strong>Vanquish UHPLC</strong></a> coupled to TSQ Altis Plus MS. The solution enables:&nbsp;</p><ul><li>Robust quantitation of semaglutide and liraglutide with a LOQ of 0.1 ng/mL&nbsp;</li><li>Excellent precision and accuracy across 3 orders of LDR for the analysis of GLP-1 analogs&nbsp;</li><li>Minimal carryover observied with post-injection signals remaining below 10% of the LOQ for semaglutide and below 14% for liraglutide following the highest concentration injection (100 ng/mL)</li></ul><h3><strong>4. Waters Corporation: Method Migration of a Reversed Phase TFAAcetonitrile Gradient Method From a Binary to a Quaternary System: Impact of Mixing</strong></h3><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/720008689_en_51b4c42ae8.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Solvent mixing is critical to obtaining optimal gradient LC separations. The most commonly available pump mixing designs for reversed phase gradient HPLC separations are high pressure (typically binary) and low pressure (quaternary) systems. High pressure systems use independent pumps to deliver different solvents. Mixing occurs at high pressures with solvent composition controlled by the flow rates of the pump heads. In low pressure systems, solvent composition is controlled by a gradient proportioning valve. Each solvent is delivered in packets (based on the gradient specified in the method) which are mixed as they go through the pump head. High pressure systems have reduced dwell volumes and generally have improved compositional accuracy (leading to better retention time precision) over low pressure systems. Both high- and low-pressure systems are subject to mobile phase composition fluctuations. Mixers of various volumes and designs are utilized to help minimize these composition fluctuations by reducing baseline noise and oscillations in mobile phase composition.&nbsp;</p><p>The use of certain mobile phase modifiers is known to impact chromatographic baselines. Trifluoroacetic acid (TFA) is a commonly used ion-pairing reagent used in combination with acetonitrile for many gradient reversed phase applications. TFA absorbs strongly at wavelengths below 250 nm. Additionally, TFA is slightly retained on reversed phase columns which results in fluctuations in TFA concentration as the acetonitrile gradient passes through the column. In combination, these factors result in significant baseline disturbances (ripples) when TFAacetonitrile gradients are used at low wavelengths. These baseline ripples may make peak integration difficult and can impact the sensitivity along with peak area precision and retention time precision of the method.&nbsp;</p><p>Most modern HPLC systems typically include a standard mixer in the pump design. Additionally, different mixers may be available to improve mixing performance for specific applications. The effect that mixers have on chromatographic performance was demonstrated during the migration of a reversed phase TFA-acetonitrile gradient method from a binary system to the quaternary Alliance iS HPLC System.</p><h4><strong>Experimental</strong></h4><ul><li><strong>LC systems: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/717"><strong>Alliance iS HPLC System</strong></a> Comparable Binary HPLC system (System X)&nbsp;</li><li><strong>Detection: </strong>UV Detector with 10 mm HPLC flow cell</li><li><strong>Column: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1823"><strong>XBridge™ C18 Column</strong></a>, 5 µm 4.6 x 250 mm (p/n: 186003117)</li></ul><h5><strong>Data Management&nbsp;</strong></h5><ul><li><strong>Chromatography data system: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/627"><strong>Empower™ 3.8.0.1</strong></a></li></ul><h4><strong>Conclusion&nbsp;</strong></h4><p>The differences in mixing performance between binary and quaternary HPLC Systems may impact method migration. Mixing performance is particularly important when migrating gradient TFA-acetonitrile methods at low wavelengths, as these methods are known to produce significant baseline noise (ripple). In this study, effect of mixers on chromatographic performance was demonstrated through the migration of the USP Tryptophan Organic Impurities method from a binary system to the quaternary Alliance iS HPLC System. The results show that the characteristic baseline ripple associated with TFA-Acetonitrile gradients at low wavelengths is impacted by the choice of mixer. The Alliance iS HPLC System with the 690 µL Ti diffusion bonded mixer displayed a significant reduction of baseline noise when compared against both the binary and quaternary systems equipped with their respective standard mixers. This resulted in easier peak integration, improved peak area and retention time reproducibility, and increased method sensitivity.</p>

News from LabRulezLCMS Library - Week 38, 2025

<p><strong>A stability-indicating RP-HPLC method was developed and validated for impurity analysis of the cystic fibrosis drug Elexacaftor using a Quality-by-Design (QbD) approach. The method achieved effective separation of six known impurities with appropriate resolution and peak shape under optimized chromatographic conditions.</strong></p><p>Validation according to ICH guidelines confirmed the method’s robustness, linearity, accuracy, specificity, and precision. Stress testing under hydrolytic, oxidative, thermal, and photolytic conditions demonstrated suitability for degradation studies, supporting both process development and routine quality control of bulk drug substances.</p><blockquote><h4><strong>The original article</strong></h4><h4><a target="_blank" rel="noopener noreferrer" href="https://pubs.acs.org/doi/10.1021/acsomega.4c10834"><strong>Quality-by-Design-Driven RP-HPLC Method Development and Validation for Impurity Analysis of Elexacaftor, a Cystic Fibrosis Drug, with LC-MS/MS-Based Degradant Identification</strong></a></h4><p>Jayaprakash Kanijam Raghupathi, Divya Kumar Vemuri, Syed Mastan Ali, Vishnuvardhana Kishore Polisetti, Rambabu Gundla, Leela Prasad Kowtharapu, Dittakavi Ramachandran*, and Naresh Kumar Katari*</p><p>ACS Omega 2025, 10, 39, 44885–44894</p><p><a target="_blank" rel="noopener noreferrer" href="https://doi.org/10.1021/acsomega.4c10834">https://doi.org/10.1021/acsomega.4c10834</a></p><h6>licensed under <a target="_blank" rel="noopener noreferrer" href="https://creativecommons.org/licenses/by/4.0/"><strong>CC-BY 4.0</strong></a></h6></blockquote><p><strong>Selected sections from the article follow. Formats and hyperlinks were adapted from the original.</strong></p><p>Elexacaftor is a drug that is used to treat fibrosis (TCF). When treating patients with cystic fibrosis (CF) who have the F508del mutation or other genetic mutations that react well with these medications, it is specifically used in conjunction with two other medications, ivacaftor and tezacaftor. The cystic fibrosis transmembrane conductance regulator, or CFTR, modulator, is Elexacaftor. Due to genetic abnormalities, people with CF have abnormal CFTR protein function, which is improved by this treatment. Based on clinical research, individuals with CF should expect a much better quality of life, reduced pulmonary exacerbations, and improved lung function when using Elexacaftor, ivacaftor, and tezacaftor together as opposed to prior treatments. After receiving FDA approval, Elexacaftor was marketed in the USA under the brand name Trikafta in October 2019. <a href="javascript:void(0);">(1−5)</a> Since then, it has emerged as a key component of the CF treatment plan, especially for those with specific mutations. It is taken orally as tablets or granules, usually once a day, along with a meal high in fat. Headache, upper respiratory tract infection, abdominal pain, diarrhea, rash, and elevated liver enzymes are among the frequently reported side effects. Generally speaking, these side effects are controllable and can be well watched during therapy.</p><p>To ensure the quality and safety of ELX, it is required to develop analytical methods for the identification and estimation of impurities. The currently available literature reveals that the ELX drug combination with ivacaftor and tezacaftor is used very effectively. So, most of the analytical methods related to the estimation of three drugs simultaneously quantitatively by using different analytical techniques, i.e., by high-performance liquid chromatography and liquid chromatography–mass spectroscopy. Several liquid chromatography-ultraviolet (LC-UV) methods <a href="javascript:void(0);">(6,7)</a> and liquid chromatography-tandem mass spectrometry (LC-MS/MS) <a href="javascript:void(0);">(8−15)</a> methods are available to simultaneously measure the concentration of different combinations of IVA/TEZ/ELX with metabolites in plasma or other biological fluids. However, we did not find the literature for the analysis of process-related impurities and degradation impurities. The methods available in the literature do not discuss the process-related impurities and the degradation impurities of the ELX drug substance; hence, a stability-indicating LC method was developed for identifying and estimating the process-related impurities and degradation impurities of the ELX drug substance. In this paper, we report on a new analytical method to the estimation of ELX and its six process-related impurities quantitatively by using an HPLC instrument. This new LC method is sensitive enough to detect impurities as low as 0.02%, which ensures there is no harm or risk to the patients from impurities; however, existing methods in the literature do not discuss the sensitivity of the potential impurities, which can harm the patients.</p><p>The method has been validated according to current international guidelines; this method is specific and rugged for analysis. An experimental design constructed according to Quality by Design (QbD) can identify the individual effects as well as interactions of the critical method variables, such as organic ratio, pH of the buffer or mobile phase, flow rate, column oven temperature, and gradient composition. <a href="javascript:void(0);">(16−18)</a> It is a systematic approach to analytical method development that is risk-based, which helps to create robust and reliable analytical methods. The current robustness method was studied by utilizing the QbD-based Design of Experiments (DoE) with Design-Expert software.</p><h4><strong>2. Materials and Methods</strong></h4><h5><strong>2.2. Instrumentation and Software</strong></h5><p><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/210"><strong>Waters Alliance e2695 HPLC</strong></a> with a quaternary pump was used for method development and analysis (34 Maple St, Milford, Massachusetts 01757, USA). Empower 3 software (Waters Corporations, USA) was used for data collection and processing. Mettler Toledo microbalance was used, on which weighing was done on an MX5 microbalance (Mettler Toledo, Columbus, Ohio, USA). A SevenMulti pH meter (Mettler Toledo Columbus, Ohio, USA) was used for estimation of the pH.</p><h5><strong>2.3. HPLC Instrumentation and Conditions</strong></h5><p>Waters Alliance’s e2695 LC system with the 2998 photodiode array (PDA) detector was used for method development, method validation, and forced degradation investigations. Using Empower software, data were gathered and processed. Using a Waters 2998 PDA detector, the peak homogeneity was investigated.</p><h5><strong>2.3.1. Chromatographic Method Conditions</strong></h5><p>The analysis of Elexacaftor employed the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1823"><strong>XBridge Shield RP18</strong></a>, 150 mm × 4.6 mm, 3.5 μm column with 10 mm KH<sub>2</sub>PO<sub>4</sub> in water, pH adjusted to 3.2 with phosphoric acid buffer, and showed positive and encouraging results. Furthermore, extensive trials were made on an XBridge shield RP column with various flow rate and gradient conditions. The optimum separation of all impurities and ELX was achieved on XBridge Shield RP18, 150 × 4.6 mm, 3.5 μm, with mobile phase A as buffer:methanol (98:2, v/v) and mobile phase B as acetonitrile:2-propanol:water (70:20:10, v/v/v), flow rate 0.7 mL/min; column temperature 27 °C; with binary gradient time (min)/B concentration (%): 0/5, 20/50, 60/85, 80/85, 82/5 and 90/5. All peaks were separated with good resolution and peak shape under these conditions and are shown in <a target="_blank" rel="noopener noreferrer" href="https://pubs.acs.org/doi/10.1021/acsomega.4c10834#fig2">Figure 2</a>. The diluent was a 40:60 v/v solution of water and acetonitrile that was also used to wash the needles.</p><h4><strong>3. Results and Discussion</strong></h4><h5><strong>3.2. Optimized Chromatographic Conditions</strong></h5><p>Based on the method developmental trials, it was found that Elexacaftor and related substances were not retained and separated as per the requirement in any of the screened conditions except the trial with XBridge Shield RP 18, 150 mm × 4.6 mm, 3.5 μm, mobile phase A as buffer:methanol (98:2, v/v) and mobile phase B as acetonitrile:2-propanol:water (70:20:10, v/v/v), flow rate 0.7 mL/min; column temperature 27 °C; with binary gradient time (min)/B conc (%): 0/5, 20/50, 60/85, 80/85, 82/5, and 90/5. All seven peaks were separated with good resolution and peak shape in this condition. The details of the finalized method conditions are mentioned in <a target="_blank" rel="noopener noreferrer" href="https://pubs.acs.org/doi/10.1021/acsomega.4c10834#tbl2">Table 2</a> for quantitating ELX and its related substances by using HPLC.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/ACS_Omega_2025_10_39_44885_44894_Table_2_Analysis_of_Optimized_HPLC_Method_Conditions_for_Elexacaftor_and_Its_Impurities_5fc5c933c5.jpg" alt="ACS Omega 2025, 10, 39, 44885–44894: Table 2. Analysis of Optimized HPLC Method Conditions for Elexacaftor and Its Impurities" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_ACS_Omega_2025_10_39_44885_44894_Table_2_Analysis_of_Optimized_HPLC_Method_Conditions_for_Elexacaftor_and_Its_Impurities_5fc5c933c5.jpg 225w," sizes="100vw" width="1046" height="726"></p><p>With the above chromatographic conditions, ELX and its related substances (Imp-A to Imp-F) got separated from each other with a resolution of more than 2.0, as shown in <a target="_blank" rel="noopener noreferrer" href="https://pubs.acs.org/doi/10.1021/acsomega.4c10834#fig3">Figure 3</a>.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/ACS_Omega_2025_10_39_44885_44894_Figure_3_Overlay_chromatogram_of_degradation_studies_b71797ad37.jpg" alt="ACS Omega 2025, 10, 39, 44885–44894: Figure 3. Overlay chromatogram of degradation studies." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_ACS_Omega_2025_10_39_44885_44894_Figure_3_Overlay_chromatogram_of_degradation_studies_b71797ad37.jpg 245w," sizes="100vw" width="2096" height="790"></p><h5><strong>3.3. Method Validation Results</strong></h5><p>According to ICH requirements, <a href="javascript:void(0);">(19−23)</a> the created approach was validated, and the results are shown in <a target="_blank" rel="noopener noreferrer" href="https://pubs.acs.org/doi/10.1021/acsomega.4c10834#tbl3">Table 3</a>. In the presence of its process contaminants, the devised HPLC technique for ELX’s specificity was assessed. ELX and Imp-A-Imp-F are well separated from one another. Utilizing a photodiode array detector and HPLC, the analysis was completed. All of the peaks of ELX and its impurities were shown to be pure when the peak purity tool was employed on them, indicating the method’s specificity because no additional impurities were detected coeluting at the same retention period.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/ACS_Omega_2025_10_39_44885_44894_Table_3_Method_Validation_Summary_of_the_Estimation_of_Elexacaftor_and_Its_Impurities_Part_1_ffb1d7d057.jpg" alt="ACS Omega 2025, 10, 39, 44885–44894: Table 3. Method Validation Summary of the Estimation of Elexacaftor and Its Impurities_Part 1" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_ACS_Omega_2025_10_39_44885_44894_Table_3_Method_Validation_Summary_of_the_Estimation_of_Elexacaftor_and_Its_Impurities_Part_1_ffb1d7d057.jpg 168w," sizes="100vw" width="1042" height="969"></p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/ACS_Omega_2025_10_39_44885_44894_Table_3_Method_Validation_Summary_of_the_Estimation_of_Elexacaftor_and_Its_Impurities_Part_2_75f03ca18f.jpg" alt="ACS Omega 2025, 10, 39, 44885–44894: Table 3. Method Validation Summary of the Estimation of Elexacaftor and Its Impurities_Part 2" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_ACS_Omega_2025_10_39_44885_44894_Table_3_Method_Validation_Summary_of_the_Estimation_of_Elexacaftor_and_Its_Impurities_Part_2_75f03ca18f.jpg 178w," sizes="100vw" width="1058" height="924"></p><h4><strong>4. Conclusions</strong></h4><p>For the measurement of Elexacaftor and its related substances (imp-A to imp-F and any other impurities), the suggested technique was optimized to give a sharp peak with a minimal tailing factor. This approach was found to be specific, accurate, linear, and economical. The approach offers a number of benefits, including cost-effectiveness, robustness, and increased sensitivity. It was the first time we developed a method to estimate the content of Elexacaftor drug alone in process development. The current technique has been demonstrated to be efficient in terms of cost and may be utilized to measure Elexacaftor and its related compounds in the bulk. ICH rules were followed in the method’s validation.</p>

Quality-by-Design-Driven RP-HPLC Method Development and Validation for Impurity Analysis of Elexacaftor, a Cystic Fibrosis Drug, with LC-MS/MS-Based Degradant Identification

<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 17th 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> Shimadzu / ASMS </strong>and application note by<strong> Waters Corporation!</strong></p><h3><strong>1. Agilent Technologies / ASMS: Quantitation of N-Nitroso Duloxetine N-Nitrosamine Drug Substance Related Impurities (NDSRI) in API and Formulation by 6475 LC/TQ</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_drug_impurities_lc_tq_tp_402_asms_2026_en_agilent_b933b9bddf.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Nitrosamine impurities are nitroso derivatives where the nitroso group is attached to an amine. The active pharmaceutical ingredient (API) derived from a class of nitrosamines are known as N-nitrosamine drugsubstance-related impurities (NDSRI). Nitrosamine impurities in APIs and drug products pose safety concerns, even in small quantities, and thus are a major concern for drug makers [1].&nbsp;</p><p>Analytical methods must be sufficiently sensitive to quantify the nitrosamine of interest at levels as low as 10% of the specification limit and, where feasible, down to ≤5% of the specification limit, for the determination of the limit of detection. LC/MS/MS is an inherently selective and highly sensitive analytical technique that is well suited for the identification and quantification of trace-level impurities at extremely low concentrations. Consequently, LC/MS/MS has been widely adopted within the pharmaceutical industry, particularly for nitrosamine analysis [2-3].&nbsp;</p><p>Therefore, manufacturers must develop and qualify a dedicated analytical method that is appropriate for the specific sample matrix and targeted NDSRIs. An LC/MS/MS method based on multiple reaction monitoring (MRM) was developed for the detection and quantification of the N-nitroso duloxetine NDSRI impurity over a concentration range of 0.5 to 40 ppb (absolute standard concentration), using an atmospheric pressure chemical ionization (APCI) source on an Agilent 6475A LC/TQ instrument.</p><h4><strong>Experimental</strong></h4><ul><li><strong>MS:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=6475"><strong>6475 Triple Quadrupole LC/MS System</strong></a></li><li><strong>Column:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=Poroshell+HPH+C18"><strong>Poroshell HPH C18</strong></a> (4.6*150mm, 2.7µm)</li></ul><h4><strong>Conclusions</strong></h4><ul><li>A highly sensitive and robust MRM LC/MS/MS method was developed for the quantitation of N-nitroso duloxetine in duloxetine hydrochloride API and finished formulation using the Agilent 6475A LC/TQ.&nbsp;</li><li>The optimized chromatographic conditions provided effective separation of the analyte from the API, placebo, and formulation matrices, ensuring reliable quantitation.&nbsp;</li><li>The calibration curve was linear over the concentration range of 0.5-40 ng/mL, with 1/x weighting. R and R2 values were observed for both to be greater than 0.99.&nbsp;</li><li>Recovery studies demonstrated efficient sample extraction, with recoveries ranging from 92-99% in API samples and 109-116% in pellet formulations at the LOQ, specification, and 200% specification levels.&nbsp;</li><li>The method also showed excellent reproducibility across all evaluated concentration levels, confirming its suitability for routine analysis.</li></ul><h3><strong>2. Shimadzu / ASMS: Non-Targeted Analysis of Per- and Polyfluoroalkyl Substances in Environmental Extracts by Microflow LC-QToF Coupled with Multi-spray ESI</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/TP_562_Non_Targeted_Analysis_of_Per_and_Polyfluoroalkyl_Substances_in_Environmental_Extracts_by_Microflow_LC_Q_To_F_Coupled_with_Multi_spray_ESI_c2a7e1d5bf.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Targeted screening methods are well established for monitoring Perand polyfluoroalkyl substances (PFAS) in environmental samples. However, PFAS comprise a large and evolving class of compounds, creating a need to screen existing, emerging, and transformation products across diverse matrices. Non-targeted Analysis (NTA) allows for the confirmation of suspected PFAS in the environment, but the sample characterization is often limited by analytical sensitivity. Microflow (0.5 – 40 µL/min) coupled with a multi-nozzle emitter enhances sensitivity by improving desolvation. When this is coupled to a high mass accuracy and resolution Quadrupole Time-ofFlight Mass Spectrometer (QToF), capable of data-dependent and independent MS/MS, PFAS characterization of unknown analytes is possible. In this study, various environmental extracts were processed and screened using NTA for PFAS compounds.</p><h4><strong>Methods</strong></h4><p>A <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=Nexera+Mikros"><strong>Shimadzu Nexera Mikros LC</strong> </a>was used to introduce the sample to a Newomics DuoESI source with an M3 emitter on a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=9050"><strong>LCMS-9050 QToF MS</strong></a> (Fig. 1). A direct injection on an octadecyl column without delay column was performed. Direct injection was chosen over trapand-elute configuration to minimize selectivity bias caused by the trap column.&nbsp;</p><h4><strong>Conclusion</strong></h4><p>With the increased sensitivity of microflow LC-QToF coupled with multi-spray ESI with DIA acquisition, Insight Explore aided in NTA PFAS analysis. When paired with Insight Discovery and Profiler, a streamlined batch data processing workflow with statistical analysis was shown to effectively handle complex environmental matrices and their analyses.</p><h3><strong>3. Thermo Fisher Scientific / ASMS: Robust and sensitive quantitation of 18 steroids in human serum using Thermo Scientific TSQ Certis Triple Quadrupole mass spectrometer</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_004707_lsms_18_steroids_tsq_certis_clinical_po004707_asms2026_na_en_244c52437b.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Accurate quantification of endogenous steroids in human serum is essential for biomonitoring physiological functions such as hormone regulation, reproduction, and immune response in clinical research laboratories. Liquid chromatography–tandem mass spectrometry (LC–MS/MS) is the gold standard for steroid analysis due to its superior sensitivity, selectivity, and multiplexing capability compared to immunoassays. However, steroid measurements remain challenging because of low endogenous concentrations and complex serum matrices. Here, we describe a robust and sensitive LC–MS/MS method for the simultaneous quantification of 18 steroids in human serum using the Thermo Scientific TSQ Certis triple quadrupole mass spectrometer (Figure 1). The method leverages advanced ion source design and fast acquisition to deliver enhanced sensitivity, stability and throughput suitable for high-volume laboratory workflows.</p><h4><strong>Materials and methods</strong></h4><h5><strong>Liquid chromatograph – mass spectrometry&nbsp;</strong></h5><p>Samples were analyzed on a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/744"><strong>Thermo Scientific Vanquish Horizon UHPLC system</strong></a> coupled to the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/2207"><strong>TSQ Certis MS</strong></a><strong> </strong>operated in selected reaction monitoring (SRM) mode. The mobile phase composition, analytical column details, LC gradient, and overlaid extracted ion chromatograms (EICs) are shown in Figure 3.&nbsp;</p><h5><strong>Data analysis&nbsp;</strong></h5><p>Data were acquired and processed using <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/77"><strong>Thermo Scientific TraceFinder software</strong></a> (v 5.2).</p><h4><strong>Conclusions&nbsp;</strong></h4><p>A sensitive and robust LC–MS/MS method was developed for simultaneous quantification of 18 steroids in human serum using the Vanquish Horizon UHPLC and TSQ Certis MS with OptaMax Plus ion source. The method demonstrated excellent linearity, accuracy, and reproducibility across a wide dynamic range. Optimization of MS conditions, particularly vaporizer temperature, significantly improved sensitivity. Overall, the workflow delivers high sensitivity, selectivity, and long-term stability, providing a reliable solution for high-throughput steroid analysis in clinical and research settings.</p><h3><strong>4. Waters Corporation: Alkaline-hydrolysis Method for PFAS in Textiles and Textiles Products by Xevo™ TQ-S micro with ACQUITY™ UPLC™ H-Class Plus System</strong></h3><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/720009443_en_0ab4914d72.pdf"><strong>Full PDF for download</strong></a></li></ul><div><div><section><div><div><div><div><div><div><h5><strong>Benefits&nbsp;</strong></h5><ul><li>Alkaline hydrolysis method (EN 17681-1:2025) used for preparing textiles, coated textiles and textile products for PFAS analysis allowed the inclusion of side-chain fluorinated polymers&nbsp;</li><li>Extraction method provided significant advantages in handling complex textile matrices by creating clear extracts free from dispersions, settleable solids and particulates&nbsp;</li><li>Protected the high-value LC-MS/MS components from potential damage&nbsp;</li><li>The method improved analytical sensitivity, reproducibility and accuracy, enabled cost-effective and timeefficient PFAS analysis maintained high analytical quality even for challenging high-matrix samples</li></ul><p>Many PFAS are linked to health concerns because of their ability to interfere with the body’s hormone systems, acting as endocrine disruptors also, linked to other toxic effects. Since textiles come into direct and prolonged contact with the skin, there is an increased risk of human exposure—especially for babies and children.4 In addition, PFAS pose significant environmental concerns. These “forever chemicals” do not readily break down and can be released during manufacturing, washing and disposal, allowing them to enter water pathways and persist in the environment. As a result, testing for PFAS has become increasingly important in the textile industry to demonstrate supply chain transparency, build consumer trust and protect brand reputation.</p><p>Synthetic fluorinated chemicals are categorized into two main groups - non-polymeric PFAS and polymeric PFAS, known for their use as water and grease repellent, heat resistant finishes in textiles, upholstery, leather, apparel, carpet and other industrial applications such as packaging, firefighting foams, non-stick cookware, etc. (Figure 1). PFAS are fluorinated substances contain at least one fully fluorinated methyl or methylene carbon atom (without any H/Cl/Br/I atom attached to it), i.e. with a few noted exceptions, any chemical with at least a perfluorinated methyl group or a perfluorinated methylene group.2 PFAS are classified as long chain and short chain based on their perfluorinated carbon chain length. Short chain fluorinated polymers encompasses a diverse range of chemistries, categorized according to their generic chemical and repeating units structures. Few examples are: derivatives of fluorotelomers, perfluoropolyethers with varying carbon chain lengths and acrylates, epoxylates, urethane, silicones of repeated units based structures.3 Polyfluoroalkyl substances such as fluorotelomer alcohols (FTOHs) contain C–H bonds, rendering them more susceptible to degradation. The ethoxylates in fluorotelomers-methacrylate break down to FTOH whereas polymeric fluorinated urethanes, acrylates break down to PFCAs (non-polymeric and polymeric side chain fluorinated polymers). In contrast, perfluoroalkyl substances such as PFOS and PFOA are extremely persistent due to their fully fluorinated carbon chains.&nbsp;</p><p>In alkaline hydrolysis methods, methanolic aqueous sodium hydroxide is used to break down precursor PFAS such as FTOHs, branched side chain PFAS. This process releases terminal perfluorinated carboxylic acids (PFCAs), which can then be measured. However, the efficiency of hydrolysis varies among different PFAS compounds and the presence of interfering substances such as organic matter or other chemicals in the sample matrix can inhibit the cleavage and detection of these analytes.</p><h4><strong>Experimental</strong></h4><ul><li><strong>LC system:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/198"><strong>ACQUITY UPLC H-Class Plus System</strong></a> and SMFTN fitted with PFAS kit</li><li><strong>Column(s):</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1823"><strong>XBridge™ BEH™ C18 5 µm Column</strong></a>, 4.6 x 100 mm&nbsp;</li><li><strong>MS system: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/209"><strong>Xevo TQ-S micro Mass Spectrometer</strong></a></li></ul><h4><strong>Conclusion&nbsp;</strong></h4><p>By strictly following the critical steps of the alkaline hydrolysis method using methanolic aqueous sodium hydroxide (NaOH) both freely extractable PFAS and those previously bound within side-chain fluorinated polymers such as esters linked to fluorotelomer alcohols (n:2 FTOHs, where n = 4, 6, 8, 10, 12), can be effectively extracted. This approach also safeguards the LC-MS/MS instrumentation from blockages, damage, or contamination by preventing polymer accumulation and matrix interferences.&nbsp;</p><p>The analytical method using the ACQUITY UPLC H-Class Plus System and Xevo TQ-S micro LC-MS/MS System was found to be sensitive, reproducible and accurate, enabling cost-effective and time-efficient PFAS analysis in chemically processed textiles, coated textiles and textile products whilst maintaining high analytical quality even for challenging high-matrix samples.</p></div></div></div></div></div></div></section></div></div>

News from LabRulezLCMS Library - Week 34, 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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Waters Corporation, the world's leading specialty measurement company, has pioneered chromatography, mass spectrometry and thermal analysis innovations serving the life, materials and food sciences for nearly 60 years.

Waters Corporation (Europe)

2026 AOAC INTERNATIONAL Annual Meeting & Exposition

Bruker at the 7th European Symposium on Single Cell Proteomics

Shimadzu at 2026 AOAC Annual Meeting & Exposition

ICP-MS for Hybrid Photoelectrode Characterization

Modern Methods for Faster Microbial Analysis

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