News from LabRulezLCMS Library - Week 32, 2025

<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 25th 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 poster by <strong>Agilent Technologies / ASMS, </strong>brochure by<strong> Shimadzu, </strong>other document by<strong> Thermo Fisher Scientific </strong>and application note by <strong>Waters Corporation!</strong></p><h3><strong>1. Agilent Technologies / ASMS: Highly Sensitive Quantitation of N-Nitroso Timolol Impurity in Timolol API using the Agilent </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=6495D+LC%2FTQ"><strong>6495D LC/TQ</strong></a></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/ps_6495d_nitrosamines_asms_2025_mp_548_en_agilent_7c9b0b32ef.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Nitrosamines are of concern as most of them have been reported to be potent mutagens in rodents and are potential carcinogens. Therefore, it is important to control these impurities at or below the stipulated specification limits. API-derived complex nitrosamines are known as nitrosamine drug substance-related impurities (NDSRIs). Multiple recalls of pharmaceutical drug products due to the presence of NDSRIs have drawn the attention of regulators and manufacturers. By considering the complexity of the global pharmaceutical supply chain, manufacturers must be more diligent to protect consumers by screening their APIs and drug products for the presence of NDSRIs. NDSRIs can be formed from both secondary and tertiary amines. Secondary amines can easily undergo nitrosation in the presence of trace amounts of acid. Timolol is beta-blocker class of drug used for hypertension. N-nitroso timolol is an API specific nitrosamine impurity. For most NDSRIs, analytical methods are not yet available, so manufacturers will need to develop, qualify, and validate a method that is suitable for the sample matrix and specific for the nitrosamines in question.</p><p>The analytical method developed must be sensitive enough to quantify the nitrosamines of interest down to a level that corresponds to 10% of specification limit.</p><p>In this poster we highlight a highly specific, sensitive, and reproducible method that was developed for the quantitation of N-nitroso Timolol impurity in Timolol drug substance using the 6495D triple quadrupole LC/MS (LC/TQ) coupled to the 1290 Infinity II LC.</p><h4><strong>Experimental</strong></h4><h5><strong>LC configurations and parameters:</strong></h5><ul><li><strong>Instrument:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=6495D+LC%2FTQ"><strong>Agilent 6495D LC/TQ</strong></a> coupled to <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</strong></a></li><li><strong>Analytical Column: </strong>Agilent Infinity Lab Poroshell PhenylHexyl, 150 × 3.0 mm, 2.7 µm, p/n, 693975-312</li></ul><h4><strong>Conclusions</strong></h4><p>A highly sensitive and robust multiple reaction monitoring (MRM) method was developed utilizing the Agilent 6495D LC/TQ to quantify N-nitroso timolol impurity in timolol API. &nbsp;</p><ul><li>The chromatographic method provided desirable separation between the impurity and the API to avoid interference.&nbsp;</li><li>The developed method demonstrated excellent linearity over the range of 5 to 1500 pg/mL (0.5 to 150 ppb with 100 mg/mL API load) with an R2 value greater than 0.9990&nbsp;</li><li>The LOD and LOQ values achieved were 2 and 5 pg/mL respectively for targeted NDSRI&nbsp;</li><li>The method provided recovery between 90 and 110%, which is within the acceptance criteria</li></ul><h3><strong>2. Shimadzu: Liquid Chromatograph Mass Spectrometer </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=LCMS-8065XE"><strong>LCMS-8065XE&nbsp;</strong></a></h3><ul><li>Brochure</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/c146_e500_526bbea8e9.pdf"><strong>Full PDF for download</strong></a></li></ul><h5><strong>Shimadzu LCMS-8065XE: Next-Generation Triple-Quadrupole Performance</strong></h5><p>Shimadzu introduces the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=LCMS-8065XE"><strong>LCMS-8065XE</strong></a>, a triple-quadrupole liquid chromatograph mass spectrometer designed to set new standards in sensitivity, speed, and robustness. Built on Shimadzu’s philosophy of “Contributing to Society Through Science and Technology,” the system integrates advanced hardware with Analytical Intelligence software, ensuring both expert-level optimization and user-friendly operation. This balance of evolved, efficient, and exact capabilities makes the LCMS-8065XE a powerful solution for modern laboratories.</p><h5><strong>Breakthrough Ionization and Detection Technologies</strong></h5><p>At the heart of the system are innovations such as the <strong>StreamFocus ESI probe</strong> and IonFocus technology, which enhance ion intake, reduce gas consumption, and eliminate contamination without compromising sensitivity. Combined with the refined <strong>UFsweeper IV collision cell,</strong> the instrument achieves exceptional signal transmission and robust long-term stability. Demonstrated by ultra-trace PFAS detection at parts-per-trillion levels, the LCMS-8065XE ensures reliable results even under continuous high-throughput conditions.</p><h5><strong>Ultra-Fast Mass Spectrometry with UF Technologies</strong></h5><p>Shimadzu’s proprietary UF technologies—including UF-Qarray II, UF-Lens II, and UF-MRM—deliver unmatched speed and sensitivity. Capable of 5 msec polarity switching, up to 555 MRM channels per second, and scanning at 30,000 u/sec, the system supports complex multi-component analyses with precision. Intelligent maintenance features, such as quick desolvation line replacement without breaking vacuum, further minimize downtime and maximize laboratory productivity.</p><h5><strong>Smart Software, Sustainability, and Seamless Integration</strong></h5><p>The LCMS-8065XE is complemented by Peakintelligence AI algorithms, LabSolutions Connect MRM, and LabSolutions Insight, enabling automated tuning, optimized methods, and reliable peak integration with reduced manual review time. Sustainability is also central: ecology mode reduces power consumption by up to 31%, while integration with Shimadzu’s Nexera UHPLC system improves throughput and reduces resource use. Features like FlowPilot smart flow control and automatic bubble detection ensure uninterrupted performance, extending system and column lifetimes while cutting operational costs.</p><h3><strong>3. Thermo Fisher Scientific: Accucore columns Resolve all your impurities without feeling the pressure</strong></h3><ul><li>Other document</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/fl_002099_ccs_lc_columns_accucore_C18_HILIC_phenyl_fl002099_na_en_db7abbabd7.pdf"><strong>Full PDF for download</strong></a></li></ul><p><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products?category=48&amp;manufacturerItems=3&amp;search=accucore"><strong>Thermo Scientific™ Accucore™ columns</strong> </a>bring UHPLC-level performance to both HPLC and UHPLC systems, even when pressure capabilities are limited. With <strong>14 distinct chemistries</strong>, they provide an extensive toolbox for successful method development and robust method transfer. Built on decades of innovation in stationary phase design, Accucore columns ensure reliable performance across diverse analytical challenges.</p><h5><strong>Comprehensive Chemistry Portfolio for Any Separation Need</strong></h5><p>The Accucore family includes multiple column chemistries tailored to different selectivity requirements. <strong>Hydrophobic options like </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/427"><strong>C18</strong></a><strong>, </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/428"><strong>C8</strong></a><strong>, and </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1990"><strong>C4</strong></a><strong> </strong>deliver classic reversed-phase separations, while <strong>polar-modified C18</strong> (<a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/430"><strong>Polar Premium</strong></a><strong>, </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/429"><strong>aQ</strong></a>) enhances retention of acidic and basic compounds. <strong>Phenyl-based phases</strong> such as <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/424"><strong>Biphenyl</strong></a><strong>, </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/431"><strong>Phenyl-Hexyl</strong></a><strong>, and </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1985"><strong>PFP</strong></a> provide orthogonal selectivity for aromatic and polar analytes, and <strong>specialty hydrophobic columns (</strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/426"><strong>RP-MS</strong></a><strong>, </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1988"><strong>C30</strong></a>) expand capabilities for structural isomer separation. For highly polar compounds, <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1987"><strong>HILIC</strong></a> and <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1989"><strong>Amide-HILIC</strong></a> chemistries enable reliable retention and resolution.</p><h5><strong>Optimized for Speed, Sensitivity, and Versatility</strong></h5><p>Engineered with advanced particle design, Accucore columns offer rapid separations with high resolution and reproducibility. Their versatility makes them suitable for a wide range of applications, from pharmaceutical impurity profiling to food and environmental analysis. By offering both routine and specialty phases, the columns allow analysts to fine-tune selectivity and maximize productivity without compromising robustness.</p><h5><strong>Trusted Innovation and Seamless Integration</strong></h5><p>As part of Thermo Fisher Scientific’s portfolio, Accucore columns are backed by extensive expertise in HPLC method development and transfer. Their design supports high-throughput workflows, smooth integration into existing systems, and consistent results across laboratories. Whether the need is for routine testing or complex separations, Accucore columns deliver dependable performance with the flexibility to resolve impurities efficiently and confidently.</p><h3><strong>4. Waters Corporation: Temperature Control Using Waters LC Purification Systems with Timberline's Column and Mobile Phase Heaters: Optimizing Preparative Performance&nbsp;</strong></h3><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/720008874_en_6514d1a04e.pdf"><strong>Full PDF for download</strong></a></li></ul><h4><strong>Benefits&nbsp;</strong></h4><ul><li>Minimizing method redevelopment at the prep scale improves process efficiency and reproducibility, ensuring predictable and reliable outcomes for isolating small and large molecules, such as oligonucleotides and peptides.&nbsp;</li><li>Elevated temperature reduces system backpressure and enables one to consider using of sub-5 µm <strong>Waters™ preparative Optimum Bed Density (OBD™) columns </strong>to increase productivity through faster separations, increased loading capacity, lower solvent consumption, shorter fraction dry-down times, and reduced waste disposal costs.&nbsp;</li><li>Enabling the substitution of acetonitrile with methanol provides a more environmentally sustainable and economically efficient purification process.</li></ul><p>Although temperature control is widely implemented in analytical HPLC to improve separations and ensure consistent and reliable outcomes, temperature is seldom controlled or used as a parameter for manipulating chromatography at the preparative scale for two reasons. First, high flow rate separations occur at the temperature of the incoming solvent and efficient heating of the mobile phase entering the column is challenging at these high flow rates. Second, large diameter columns cannot be effectively heated from the outside simply by using insulation, heated blankets or ovens. Ideally, analytical separations developed with temperature control should be scaled to prep at the same temperature to eliminate variability in the chromatography which can make the target identification and isolation ambiguous or lead to product loss.&nbsp;</p><p>When used with the Waters AutoPurification System, Timberline’s column and mobile phase heaters provide precise and consistent temperature control for preparative separations, allowing easier scale-up from UPLC/UHPLC to preparative methods. This integrated approach minimizes the need for method redevelopment, improves reproducibility, enhances hydrophobic compound solubility and boosts operational efficiency, which is particularly valuable in small molecule isolations and biological applications such as oligonucleotide and peptide purification. Temperature control also enhances flexibility in purification workflows, supporting green chemistry initiatives by enabling the substitution of acetonitrile with methanol. Elevated temperature reduces system backpressure, allowing the usage of sub-5 µm particles in larger-dimension preparative columns (e.g., 10 mm ID), which may increase loading capacity, minimize run-times, reduce solvent consumption, shorten dry-down times and lower waste disposal costs. This technical brief illustrates that comparable purification results can be achieved using methanol with sub-5 µm 10 mm ID columns, an approach that is both environmentally sustainable and economically viable. It also illustrates the precision and stability of temperature control across various flow rates and mobile phase compositions, showcasing the reliability of this solution.</p><h4><strong>Conclusion&nbsp;</strong></h4><p><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=5&amp;search=Timberline%E2%80%99s+TL-105+D+column"><strong>Timberline’s TL-105 D column</strong></a> oven configured with the Waters AutoPurification System provides precise and consistent temperature control when scaling from analytical to preparative methods. This significantly reduces the need for method redevelopment at the prep scale, enhances chromatographic efficiency and reproducibility and delivers predictable and reliable outcomes for various applications including small and large molecules such as oligonucleotides and peptides isolations.</p><p>This combined solution supports the growing demand for green chemistry by enabling the substitution of acetonitrile with methanol. Enhanced temperature control also lowers system backpressure, allowing the use of smaller particle size, larger dimension Waters preparative OBD columns. This results in faster separations, increased loading capacity, reduced solvent consumption, shorter fraction dry-down times, and lower waste disposal costs. These benefits contribute to a more environmentally sustainable and economically efficient purification process.</p>

News from LabRulezLCMS Library - Week 35, 2025

<p><strong>Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 1st 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 note by <strong>Agilent Technologies, </strong>posters by<strong> Shimadzu </strong>and <strong>Waters Corporation </strong>and<strong> </strong>other document by<strong> Thermo Fisher Scientific!</strong></p><h3><strong>1. Agilent Technologies: PFAS Analysis in Human Plasma Using the Agilent Ultivo Triple Quadrupole LC/MS&nbsp;</strong></h3><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/an_pfas_ultivo_tq_lcms_5994_8487en_agilent_2968eb7f88.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Perfluoroalkyl and polyfluoroalkyl substances (PFAS) are a group of synthetic compounds that have been widely used in various industrial and consumer applications, including cosmetics and personal care products. Due to their widespread use, persistence in the environment, and known bioaccumulation with adverse effects, PFAS are a significant concern for human health. For example, PFAS have been linked to several adverse health effects, including cardiometabolic diseases,<sup>1</sup> reproductive effects,<sup>2</sup> and increased risk of certain cancers.<sup>3</sup> PFAS exposure has been shown to lead to alterations in lipid and energy metabolism, contributing to metabolic dysfunction.<sup>4,5</sup> It is also known that PFAS accumulate in liver and other tissues, contributing to age‑related morbidities.<sup>6</sup> To increase the understanding the molecular basis of these effects, analysis of PFAS in blood plasma has been combined with lipidomics to identify the lipid biomarkers associated with metabolic risks.<sup>7</sup> Further, the PFAS concentrations in serum have also been shown to differ by gender<sup>7,8 </sup>and ethnicity.<sup>9</sup> Considering these and other findings, monitoring PFAS levels in plasma is important to understand their health impacts and develop strategies for their mitigation. This application note validates an LC/MS/MS method for targeted quantification of fourteen PFAS in plasma using the Agilent Ultivo Triple Quadrupole (TQ) LC/MS. For the method presented, the Ultivo LC/TQ offers performances on par with mid- to high-end TQ systems in a much smaller footprint. The protein-precipitation-based sample preparation method is fast and effective for plasma samples. Method performance in terms of sensitivity, linearity, carryover, accuracy, and precision are described. Stability of targeted PFAS in quality control (QC) samples and analysis of real human plasma samples are also discussed.</p><h4><strong>Experimental</strong></h4><h5><strong>Instrumentation and LC/MS/MS method&nbsp;</strong></h5><p>Chromatographic conditions were optimized for analyte separation and retention time for each of the PFAS as determined by analyzing the labeled and non-labeled standards. PFAS were separated by reversed-phase LC on an Agilent InfinityLab Poroshell 120 EC‑C18, 2.1 × 50&nbsp;mm, 2.7 μm (part number 699775-902) and detected in negative electrospray ionization (ESI) mode using an <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=1290+Infinity%C2%A0II+LC"><strong>Agilent 1290 Infinity&nbsp;II LC</strong></a> system coupled with the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=Ultivo+LC%2FTQ"><strong>Ultivo LC/TQ</strong></a>. The liquid chromatograph was equipped with a binary pump (part number G7120A), column compartment (part number G7116B), and Agilent 1290 Infinity II Multisampler (part&nbsp;number G7167B). For each experiment, 5 μL of extract was injected. The LC parameters are listed in Table 2. The Ultivo LC/TQ was operated in dynamic multiple reaction monitoring (dMRM) mode. The dMRM parameters and retention times were optimized by analyzing each unlabeled PFAS standard. The MS parameters are listed in Table&nbsp;3. The Agilent <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=MassHunter+Acquisition+software"><strong>MassHunter Acquisition software</strong></a> dMRM parameters per PFAS are in Table 4. Agilent <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=MassHunter+Quantitative+Analysis+software"><strong>MassHunter Quantitative Analysis software </strong></a>was used for data processing.</p><h4><strong>Conclusion&nbsp;</strong></h4><p>The quantitative method for the analysis of 14 PFAS in human plasma samples that is presented here is simple and robust, uses relatively small plasma volumes, and relies on fast and effective protein precipitation, facilitating its large-scale application. The method's sensitivity, linearity, carryover, recovery, and precision make it scalable to population-based studies and biomonitoring. The compact, stackable Ultivo LC/TQ and MassHunter software, in combination with columns and consumables from Agilent Technologies, offer a fit-for-purpose, end-to-end solution to enable widespread adoption in both routine monitoring and research. The method has already been used in combination with plasma lipidomics to characterize the relationship of circulating PFAS and lipidomic profiles.7 These types of studies promise to reveal the molecular basis of the adverse effect of PFAS exposure on human health&nbsp;outcomes.</p><h3><strong>2. Shimadzu / ASMS: One System, Multiple Solutions: Analysis of PFAS &amp; Cyanotoxins in Water Adhering to EPA 537.1, 544, and 545&nbsp;</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/ASMS_25_WP_134_197d4ffd22.pdf"><strong>Full PDF for download</strong></a></li><li>PFAS are persistent synthetic chemicals found in consumer and industrial products, while cyanotoxins are harmful substances produced by cyanobacteria in water bodies.&nbsp;</li><li>EPA Methods 537.1 and 533 are approved by EPA for PFAS analysis in drinking water; Methods 544 and 545 target cyanotoxins like microcystins, nodularin, cylindrospermopsin, and anatoxin-a.&nbsp;</li><li>A single Shimadzu <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=LCMS-8060RX"><strong>LCMS-8060RX</strong></a> system with automatic method switching enables efficient, accurate analysis of PFAS and cyanotoxins, reducing equipment needs and turnaround time.</li></ul><h4><strong>2. Methods</strong></h4><p>Figure 1 illustrates the LC-MS system’s method-switching capability, enabling seamless transitions between EPA Methods 537.1, 544, and 545 on a single platform. For EPA Method 537.1 (PFAS analysis), the flow path includes the delay column to remove background PFAS. In contrast, for EPA Methods 544 and 545 (cyanotoxin analysis), the delay column is bypassed to prevent unnecessary contaminations, ensuring accurate results. This automation enhances workflow efficiency and analytical flexibility.&nbsp;</p><p>To prevent mobile phase contamination when switching between PFAS and cyanotoxin analysis, a simple five-minute rinse (the flow path indicated by the red lines in Figure 2) with the appropriate mobile phase is shown to be sufficient.</p><h4><strong>4. Conclusion</strong></h4><p>This study shows that PFAS and cyanotoxins can be accurately measured using one triple quadrupole mass spectrometer with automatic method switching. A quick five-minute rinse ensures clean transitions between methods, minimizing downtime and manual work. This streamlined setup reduces equipment costs, boosts efficiency, and supports high-throughput environmental testing, including rapid response to events like harmful algal blooms.&nbsp;</p><h3><strong>3. Thermo Fisher Scientific: Nestlé develops an </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/401"><strong>Orbitrap Exploris 480 mass spectrometer</strong></a><strong> method for wide-scope analysis of pesticides and natural toxins in foods</strong></h3><ul><li>Other document (Case study)</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/cs_003406_lsms_nestle_orbitrap_exploris_480_case_study_cs003406_na_en_790e517c7d.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Nestlé is committed to ensuring the highest standards of food quality and safety through rigorous research and innovation. Located in Lausanne, Switzerland, Nestlé Research plays a pivotal role in developing analytical methods for monitoring chemical contaminants in food products. These robust methodologies are implemented across Nestlé's global laboratory network to screen, identify, and ensure compliance with food safety standards. In response to the challenges posed by climate change, evolving regulations, and shifting consumer preferences, Nestlé Research has developed a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/421"><strong>QuEChERS</strong></a>-based liquid chromatography high-resolution mass spectrometry (LC-HRMS) workflow. This innovative approach allows for the simultaneous analysis of over 1,100 contaminants, including pesticide residues, mycotoxins, and plant toxins, across various food matrices such as cereals, fruits, and vegetables. The method's sensitivity and low quantification limits ensure compliance with regulatory standards while facilitating the exploration of additional compounds to preemptively address future food safety risks.</p><h4><strong>Technology selection&nbsp;</strong></h4><p>Next, Nestlé Research scientists aimed to identify the most suitable technology to address the challenges associated with broad-scope contaminant analysis. The versatility and high selectivity of LC-HRMS for screening a broad range of compounds offered a solution. Theoretically, in full-scan mode, HRMS instruments can monitor an unlimited number of molecules simultaneously, opening up avenues for continuous extension of the method’s scope. The high-quality, high-resolution accuratemass (HRAM) data provided by the Thermo Scientific™ Orbitrap™ mass analyzer can help differentiate between compounds with similar masses and isobaric interferences from coextracted matrix components, reducing the chance of false positives. Highresolution mass spectrometry (HRMS) systems offer the capability to perform nontargeted or retrospective analyses by leveraging existing data and potentially identify novel contaminants.&nbsp;</p><p>Due to its sensitivity and HRAM data quality, the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/401"><strong>Thermo Scientific™ Orbitrap Exploris™ 480 mass spectrometer</strong></a> was evaluated and ultimately selected for workflow development. The developed method uses the full-scan MS/data-dependent MS2 (FS-ddMS2 ) acquisition mode, which combines a full scan with MS2 scans on the targets of interest measured in the full scan.&nbsp;</p><h4><strong>Necessary compromises&nbsp;</strong></h4><p>Though HRMS technology theoretically offers an unlimited scope of analysis, the true scope of analysis remains limited by the sample preparation method, chromatographic conditions, and MS ionization efficiency. According to Bessaire, “We needed to compromise on sample preparation and extraction, the LC method—the column and the gradient—and MS ionization mode because not all organic molecules can be analyzed using a single ionization mode. The scope of compounds that we dreamt about at the beginning was not the same as what we had at the end.”&nbsp;</p><p>With these compromises in mind, a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/421"><strong>QuEChERS-based sample preparation</strong></a> followed by dispersive solid-phase extraction (dSPE) cleanup was chosen due to its effectiveness for a wide range of sample types and analyte chemistries. LC method development was carried out on the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/744"><strong>Thermo Scientific™ Vanquish™ Horizon UPHLC system</strong></a>. A 22 minute LC run using acidic mobile phases on a 15 cm reversed phase (RP) C18 column with a smooth gradient slope was chosen to retain a broad range of chemicals with various polarities, while separating a maximum number of isomers and isobaric compounds within an acceptable run time. The generic sample preparation approach has advantages as Dr. Flavia Nagy (-Badoud), Scientist, Nestlé Research noted, “With the generic multiresidue method, we can reduce the use of solvents and work towards greener chemistry.”</p><h4><strong>In-house HRAM mass spectral library maximizes compound identification&nbsp;</strong></h4><p>To maximize the method’s capability to unequivocally confirm the presence or absence of numerous compounds from a single injection, the group developed an in-house HRAM mass spectral library populated in <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1760"><strong>Thermo Scientific™ mzVault™ software</strong></a>. Creating the library was time-consuming and involved injection and analysis of individual standard solutions for each compound. Dr. Nicolas Christinat, Scientist, Chemical Contaminants, Nestlé Research described the library creation process, “We relied on analytical standards for each molecule. That’s a lot of individual injections and we acquired multiple parameters for each, including different fragments at different collision energies, at different polarities, and for different adducts. All this information was stored in our mzVault spectral library, and we can use that information to build targeted methods that use the most relevant collision energy, most abundant ions, and the best polarity. Bessaire added, “Our internal mass spectral library was key to maximizing the identification capabilities of our method.”</p><h4><strong>Orbitrap Exploris 480 mass spectrometer essential capabilities&nbsp;</strong></h4><p>The Orbitrap Exploris 480 mass spectrometer platform provides capabilities essential to developing methods that provide reliable identification and quantification.&nbsp;</p><h5><strong>Sensitivity&nbsp;</strong></h5><p>First, the platform offers good sensitivity compared to other HRMS instruments. Aurélien Desmarchelier, Scientist, Nestlé Research explained, “We need state-of-the-art instruments when we develop new methods. We evaluated the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/402"><strong>Orbitrap Exploris 240 </strong></a>and Orbitrap Exploris 480 mass spectrometers. The Orbitrap Exploris 480 system’s data pointed to a bit more sensitivity and that’s why we selected and are recommending it to the laboratories that will be implementing the method, so we are aligned.” Dr. Nagy (-Badoud) added, “Contaminant analyses are always at low levels—parts per billion normally—so it is very important that we have a sensitive instrument. That’s mostly why in the past HRMS was not considered and contaminant analysis was done by triple quadrupole instruments with targeted approaches. One of the most important criteria was that we reached the required sensitivity.”</p><h3><strong>4. Waters Corporation / ASMS: Analysis of Amino Acids in Plant-Based Proteins and Pet Foods – Modification of AOAC 2018.06 to Fit for Novel Foods and Ingredients</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/720008914_e210a444bc.pdf"><strong>Full PDF for download</strong></a></li></ul><p>The AOAC International Official Method 2018.06 specifies an analytical procedure for the analysis of amino acids (AA) in infant formula, adult nutrition and dairy products. This procedure is based on the pre-column derivatization with 6-aminoquinoline succinic carbamate (AQC) and reversed-phase ultra-performance liquid chromatography with UV detection.</p><p>When applied to pet foods, a limitation was observed in the target AA list. Specifically, <strong>hydroxyproline (HyPro)</strong>, common in pet foods, exhibited poor retention on the column, making it unmeasurable. HyPro is a marker for potential food adulteration using hydrolyzed animal products to artificially boost protein levels in plant-based proteins. It is also an indicator for animal-derived ingredients such as gelatin and collagen, which are critical in halal and kosher food testing.</p><p>Therefore, it is necessary to develop an analytical method capable of effectively separating and detecting HyPro alongside other amino acids.</p><h4><strong>Experimental</strong></h4><h5><strong>LC Conditions</strong></h5><ul><li><strong>System</strong>: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/191"><strong>ACQUITY Premier System</strong></a> (BSM) with PDA detector</li><li><strong>Detection</strong>: UV (260 nm)</li><li><strong>Tubing</strong>: 0.004 mm ID, 10.5 in (Waters p/n 430001784) between column and detector</li><li><strong>Software</strong>: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/627"><strong>Empower CDS</strong></a></li><li><strong>Columns</strong>:<ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1952"><strong>ACQUITY UPLC BEH C18</strong></a>, 1.7 μm, 2.1 mm × 150 mm (Waters p/n 186002353)</li><li>AccQ∙Tag Ultra C18, 1.7 μm, 2.1 mm × 150 mm (Waters p/n 186009954)</li></ul></li></ul><h4><strong>Conclusion</strong></h4><ul><li>Excellent separation resolution for all amino acids with high repeatability and intermediate reproducibility in RT</li><li>Amino acids in six pet foods and four common plant-based proteins successfully analyzed</li><li>Modified AOAC Method provides a reliable and practical solution for AA analysis in pet foods and plant-based proteins</li></ul>

News from LabRulezLCMS Library - Week 36, 2025

<p><strong>Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 8th 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</strong> and<strong> Shimadzu </strong>and<strong> </strong>posters by <strong>Thermo Fisher Scientific / HPLC Symposium </strong>and <strong>Waters Corporation!</strong></p><h3><strong>1. Agilent Technologies: Automated Solid Phase Extraction of PFAS from Aqueous Samples</strong></h3><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/an_1633_automated_spe_wax_carbon_s_5994_8289en_agilent_1fc425207e.pdf"><strong>Full PDF for download</strong></a></li></ul><p>In this study, three types of dual-phase SPE cartridges were evaluated based on the quality control requirements for aqueous matrices as specified in US EPA Method 1633 using automated SPE. The cartridges contained PFAS WAX and Carbon S for PFAS. In the first configuration, a 200 mg bed of PFAS WAX was layered on top of a 50&nbsp;mg bed of Carbon S with the sorbents separated by a frit. In the second configuration, 200 mg of PFAS WAX was mixed with 50 mg of Carbon S before cartridge packing. In the third configuration, 200 mg of PFAS WAX and 10&nbsp;mg of Carbon S were mixed before cartridge packing. All three configurations were evaluated under identical protocols using automated SPE and were compared to results obtained using the manual addition of carbon and separate WAX extraction. The blended sorbent containing 200 mg of PFAS WAX and 10 mg of Carbon S was selected for validation and further evaluation of several environmental matrices including groundwater from residential private wells, effluent from a wastewater storage lagoon, and a landfill monitoring&nbsp;well.</p><h4><strong>Experimental</strong></h4><h5><strong>Equipment and materials</strong></h5><p>The three Agilent dual-phase cartridges used in the study are listed in Table 1. As a performance benchmark, a single‑phase <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?search=Bond+Elut+PFAS+WAX"><strong>Bond Elut PFAS WAX sorbent</strong></a> cartridge (Agilent, part number 5610‑2151) was used with either Carbon S bulk (Agilent, part number 5610-2095) or Superclean ENVI-Carb (Millipore Sigma, part number 57210-U).</p><p>Samples were extracted using the PromoChrom eight‑channel automated SPE extractors with upside‑down shaker racks (part number SPE-03 with MOD004), anticlogging frits (part&nbsp;number CF-06), high-capacity inline filters (part&nbsp;number&nbsp;F-HC-30), and anticlogging tips (part&nbsp;number F-T-4). Sample analysis was performed using an <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?search=1290+Infinity+II+LC"><strong>Agilent 1290 Infinity II LC</strong></a> system consisting of an Agilent 1290 Infinity&nbsp;II high-speed pump (G7120A), an Agilent 1290 Infinity II multisampler (G7167B), and an Agilent 1290 Infinity II multicolumn thermostat (G7116B). The LC system was modified for PFAS analysis using the Agilent <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?search=InfinityLab+PFC%E2%80%91free+HPLC+conversion+kit"><strong>InfinityLab PFC‑free HPLC conversion kit </strong></a>(part number 5004-0006). The LC system was coupled to an <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?search=6495C"><strong>Agilent 6495C triple quadrupole LC/MS</strong></a> equipped with an Agilent Jet Stream technology ion source. Agilent <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?search=MassHunter+Workstation"><strong>MassHunter Workstation software</strong></a> was used for data acquisition (version 10.1) and <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?search=Quantitative+Analysis+"><strong>Quantitative Analysis </strong></a>for QQQ (version 12.1) was used for data analysis. Instrumental conditions were determined based on detailed multiparametric optimization experiments developed in‑house.&nbsp;</p><h4><strong>Conclusion&nbsp;</strong></h4><p>The adoption of automated solid phase extraction (SPE) for PFAS analysis in environmental matrices significantly enhances laboratory efficiency and reduces the potential for human error. Our evaluation of five extraction protocols revealed that the blended sorbent containing 200 mg of PFAS WAX and 10 mg of Carbon S provided the best performance in terms of recovery, matrix interference reduction, and fewer outliers. This configuration was successfully applied to various environmental samples, demonstrating its versatility and reliability. The findings support the use of automated SPE with optimized sorbent cartridges as a robust method for PFAS analysis, aligning with the quality control metrics of US EPA Method 1633. Future work should focus on further refining automated SPE techniques and exploring their application to other environmental matrices.</p><h3><strong>2. Shimadzu: Automated Preparation of Calibration Curves Using the Autosampler's Dilution Function</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_00914_en_089a8c34fa.pdf"><strong>Full PDF for download</strong></a></li></ul><h5><strong>User Benefits</strong></h5><ul><li>The auto-dilution function equipped in the autosampler eliminates the need for manual dilution during calibration curve creation and sample preparation.</li><li>Samples can be automatically diluted to create a calibration curve by specifying the desired dilution factor in the batch table.</li><li>Because the same method file can be used across different dilution factors, only minor adjustments are needed when changing dilution conditions.&nbsp;</li></ul><p>In LC quantification, the dilution ofstandard and sample solutions is typically performed manually using pipettes, which is a laborintensive and time-consuming process. Recent advancements have called for automation to reduce labor and improve efficiency. By automating the dilution process, workflows can be streamlined, leading to increased productivity. The Nexera series autosamplers are equipped with an automatic pretreatment function that enables automated dilution. Using the auto-dilution function, samples can be automatically prepared at any desired dilution factor and directly introduced into the analytical column. This article presents a method to automate the creation of calibration curves and subsequent quantification workflows using the autosampler’s auto-dilution function.</p><h4><strong>Automated Dilution Analysis of Organic Acid Standard Solution</strong></h4><p>Automated dilution analysis was conducted using a 1000 mg/L mixed standard solution of ten organic acids. The analytical conditions are shown in Table 2 (pretreatment program : Table 1). 1.0 mL polypropylene vials were used as mixing vials, and septum-sealed vials were used for the undiluted standard solution. Rinse solution (ultrapure water) was used as the diluent. Fig. 3 shows the chromatogram of the 100-fold diluted organic acid mixed standard solution (concentration after auto-dilution : 10 mg/L for each organic acid). The <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=Nexera+Organic+Acid+Analysis+System"><strong>Nexera Organic Acid Analysis System</strong></a> (pH-buffered electric conductivity detection) was employed for this analysis.</p><h4><strong>Conclusion</strong></h4><p>By utilizing the auto-dilution function in the autosampler, the standard undiluted solution for calibration curve creation can be automatically diluted at any desired dilution ratio. This eliminates the need for manual sample dilution and streamlines the calibration curve creation workflow. Additionally, since the autodilution function can also be applied to sample solutions, only the preparation of undiluted standard and sample solutions is required, allowing for a fully automated process from calibration curve creation to quantitation. As a result, the reduction in pretreatment work contributes to improved work efficiency and productivity.</p><h3><strong>3. Thermo Fisher Scientific / HPLC Symposium: Wide-ranging polynucleotide separation capabilities using Reversed Phase particles with variable pore geometry</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_wide_ranging_polynucleotide_separation_capabilities_po_en_1fdaba9e08.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Polynucleotides such as DNA and RNA serve many uses in the areas of basic research, industrial manufacturing, therapeutics to up/down regulate cellular functions, and as therapeutic and vaccine intermediates (i.e., as an instructional template to produce proteins that 1) supplement/replace ineffective proteins or 2) function as antigens). CRISPR technology using guide RNA (gRNA, ~100mer) to directly modify cellular chromosomes was recently leveraged to create a customized treatment within 6-months for an infant with CPS1 deficiency1. This approach has provided long-term, life-saving enzyme production negating the need for daily treatments. Associated DNA and RNA such as gRNA and mRNA (1000+mer) require detailed characterization to ensure safety and efficacy (e.g., of sufficient quantity and fidelity in delivery vehicles such as lipid nanoparticles). Here, we present a RP particle with complex pore geometry and the effects of mobile phase and temperature for separating DNA/RNA with therapeutic benefits (siRNA-gRNA-mRNA:~ 20-100- 1000+mer). Semi-prep column separations, common for sample recovery for subsequent analyses or sample enrichment for in vitro or in vivo use, is also shown.</p><h4><strong>Materials and methods</strong></h4><h5><strong>Test method(s)&nbsp;</strong></h5><p><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=3&amp;search=DNAPac+RP+column"><strong>DNAPac RP columns</strong></a> packed with 4 µm DNAPac RP media include 2.1 × 50 mm (P/N 088924), 2.1 × 50 mm (P/N 088924), and 21.2 × 150 mm (P/N 080922- 1521232). For instruments, mobile phases, gradient conditions, and other chromatographic operating parameters (flow rate, temperature, injection volume, etc.), please reference the figures and associated text. For all analyses, samples were detected by UV absorbance at 260nm. For 21.2x150mm semi-prep separations, a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products?category=40&amp;manufacturerItems=3"><strong>conventional (U)HPLC chromatography system</strong></a> with a binary pump, autosampler and detector without column compartment (ambient temperature separations) was used for all analyses. Ion Pairing Agents (Cation Anion): TEAA – Triethylamine Acetate, HAA – Hexylamine Acetate, HFIP – Hexafluoroisopropanol.&nbsp;</p><h5><strong>Data analysis&nbsp;</strong></h5><p>The <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/76"><strong>Thermo Scientific Chromeleon </strong></a>7.2.10 or 7.3 Chromatography Data Systems (CDS) software was used for data acquisition and analysis.</p><h4><strong>Results&nbsp;</strong></h4><h5><strong>DNAPac RP particle characterization – chemistry and pore structure&nbsp;</strong></h5><p>Figure 1 summarizes the chemical and physical properties of the resin, which is based on polymeric divinylbenzene (top left). Unlike conventional silica particles, the polymeric resin is not susceptible to hydrolytic degradation at neither extreme pH (1-14) nor high temperature (90+oC). The aromatic groups also provide π- π bonding with aromatic nucleotides giving different selectivity from conventional alkyl phases incapable of such interactions. The SEM image visually shows the highly variated porous structure of the particles. Barrett-Joyner-Halenda (BJH) N2 adsorption analysis details the surface area contributions of the different pore sizes. From this analysis it is clear that a continuum of pores from 50Å to 2500Å (suitable for small molecules up to macromolecule adsorption, respectively) contributes to the overall surface area of the particle. Based on the complex geometry and resulting pore structure, we commonly refer to the media as a supermacroporous (SMP) particle.</p><h4><strong>Conclusions&nbsp;</strong></h4><ul><li>High resolution separation of nucleic acids varying in length from 10 to 10,000 nucleotides is achieved using an SMP particle with a variable pore structure.</li><li>Polymeric SMP particles are compatible with different mobile phase IP compositions, high pH and high separation temperature, which are critical parameters for optimizing the separation of polynucleotides&nbsp;</li><li>SMP particles provide high resolution separations in both analytical and semi-prep formats</li></ul><h3><strong>4. Waters Corporation / ASMS: Detection and Characterization of PFAS Molecules Using Inclusion and Exclusion Features of Data Directed Analysis (DDA)</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/720008869_fb6d27e0a0.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Polyfluorinated alkyl substances (PFAS) are bioaccumulative and suspected to contribute to increased cancer risk in humans. Monitoring PFAS in biofluids is possible, but their very low concentrations present analytical challenges.</p><p>Data-Directed Acquisition (DDA) isolates ions that meet defined criteria, fragments them at elevated collision energies, and generates product ion spectra. Because PFAS ions show a characteristic <strong>mass defect</strong> due to fluorine atoms, inclusion lists can enhance their detection. Conversely, exclusion lists derived from background matrix analyses help remove interferences, simplifying ion selection for fragmentation.</p><p>Preliminary tests with mixed small-molecule samples demonstrated the utility of these lists before applying them to PFAS, where inclusion lists contained hundreds of entries and exclusion lists thousands.</p><h4><strong>Methods</strong></h4><ul><li><strong>Instrument:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/202"><strong>SELECT SERIES™ Cyclic™ Ion Mobility Mass Spectrometer</strong></a></li><li><strong>Acquisition:</strong> ToF-ESI negative DDA mode, 100 ms survey/MSMS scan times, collision energy ramps optimized by PFAS class</li><li><strong>LC System:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/191"><strong>ACQUITY Premier </strong></a>with PFAS Kit and Atlantis Premier BEH C18 AX Isolator Column (2.1 × 50 mm, 5 µm)</li><li><strong>Analytical Column:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1952"><strong>ACQUITY UPLC HSS T3 C18</strong></a> (100 × 2.1 mm, 1.8 µm) at 35 °C</li><li><strong>Flow Rate:</strong> 0.3 mL/min; <strong>Injection Volume:</strong> 5 µL; <strong>Autosampler Temp:</strong> 6 °C</li><li><strong>Mobile Phases:</strong><ul><li>A: H₂O:MeOH (95:5) with 2 mM ammonium acetate</li><li>B: MeOH with 2 mM ammonium acetate</li></ul></li><li><strong>Samples:</strong> LCMS QC mixture, native PFAS mixture (PFAC30PAR, Wellington Labs), and human serum</li><li><strong>Software:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/635"><strong>MassLynx™ 4.2 for acquisition</strong></a>, <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1544"><strong>waters_connect™ with PFAS library</strong></a> (130 entries) for processing, Tibco Spotfire® for visualization</li></ul><h4><strong>Conclusion</strong></h4><ul><li>ToF-DDA mode on a cyclic ion mobility mass spectrometer is effective for PFAS analysis.</li><li>Inclusion lists based on PFAS mass defect improve real-time detection and intelligent switching.</li><li>Background-derived exclusion lists (up to 9000 entries) further reduce interferences and enhance specificity.</li></ul>

News from LabRulezLCMS Library - Week 37, 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>Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 22nd 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: Quantitation of N-Nitroso Nebivolol in Nebivolol Tablet Formulation</strong></h3><p><strong>Using an </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=6495D+triple+quadrupole"><strong>Agilent 6495D triple quadrupole LC/MS system</strong></a></p><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/an_nebivolol_tablet_formulation_6495_qqq_lc_ms_system_5994_8053en_agilent_e7f21eb12f.pdf"><strong>Full PDF for download</strong></a></li></ul><p>N-Nitroso Nebivolol is a NDSRI of Nebivolol, a beta-1 adrenergic receptor antagonist commonly used in the treatment of hypertension and heart failure. In the carcinogenic potency categorization published by the FDA, N-Nitroso Nebivolol has been categorized as Class 4, indicating the maximum allowable intake limit for the impurity to be 1,500 ng/day. Nebivolol has maximum daily dosage limit of 40&nbsp;mg for adults.</p><p>Considering a maximum daily dose of 40&nbsp;mg and allowable intake of 1,500&nbsp;ng/day for the nitrosamine impurity in the formulation under study, the specification limit of N-Nitroso Nebivolol has been calculated as 22.5 ng/mL for the 0.6 mg/mL test concentration used during this study.&nbsp;</p><p>This application note demonstrates a highly specific, sensitive, and reproducible method for the quantitation of N-Nitroso Nebivolol in a 20 mg Nebivolol tablet formulation. The method uses a 1290 Infinity II LC coupled to a 6495D triple quadrupole LC/MS/MS. The multiple reaction monitoring (MRM) conditions and ion source parameters were automatically optimized using the optimizer tool available with MassHunter Acquisition software to maximize the sensitivity of the target analyte. The API peaks were diverted to waste with the help of an integrated, software-controlled diverter valve during quantitation of the&nbsp;impurity.</p><h4><strong>Experimental</strong></h4><h5><strong>Equipment&nbsp;</strong></h5><p>The analysis was performed on a <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> coupled to a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=6495D+triple+quadrupole"><strong>6495D triple quadrupole LC/MS/ MS</strong></a>. The LC system was equipped with the following&nbsp;modules: – Agilent 1290 Infinity II high-speed pump (G7120A) – Agilent 1290 Infinity II multisampler (G7167B) – Agilent 1290 Infinity II multicolumn thermostat (G7116B) – Agilent 1290 Infinity II diode array detector (G7117A)</p><h4><strong>Conclusion&nbsp;</strong></h4><p>A highly sensitive and robust MRM method was developed for the quantitation of N-Nitroso Nebivolol in 20&nbsp;mg Nebivolol tablet formulation using an Agilent 6495D triple quadrupole LC/MS system. The chromatographic method developed provided good separation between the analyte and the API. The method showed LOD and LOQ of 2 and 5 pg/mL, respectively. The minimum signal-to-noise ratio at the LOQ level was found to be more than 50:1 (Peak to Peak). The calibration curve from 5 pg/mL to 50&nbsp;ng/mL was found to be linear, with 1/x weighting. R and R2 values were 0.999 and 0.997, respectively. Spike recovery analysis showed the efficiency of sample extraction with a recovery percentage of 116.8% in API, 108.3% in placebo, and 111.8% in tablet formulation at the spiking level of 200 pg/mL. The method was found to be highly reproducible at the LOQ level, while the %RSD value for the area response of seven replicate injections was approximately 3.1%.</p><h3><strong>2. Shimadzu: Efficient Method Development for Synthetic Peptide and Related Impurities</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_00780_en_4d0aa014ea.pdf"><strong>Full PDF for download</strong></a></li></ul><h5><strong>User Benefits</strong></h5><ul><li>LabSolutions MD improves the efficiency of the entire workflow for method development of synthetic peptide and impurities.&nbsp;</li><li>Screening for multiple mobile phases and columns can be automated using mobile phase and column switching valves.&nbsp;</li><li>Molecular weights of peptides and related impurities can be estimated and accurately tracked with LCMS-2050, a single quadrupole mass spectrometer.</li></ul><p>Peptide therapeutics, characterized by specific amino acid sequences crucial to their function, can be synthesized chemically, similar to small molecule drugs. The production of synthetic peptides involves multiple steps, including deprotection, activation, coupling, and the cleavage of the final sequence from the solid support. Impurities, such as those resulting from premature chain termination or missing amino acids, can affect the safety and efficacy of the final product. Therefore, these impurities must be separated by liquid chromatography (LC). In LC analysis, selecting the appropriate mobile phase and column across a wide range of combinations is critical for achieving optimal separation, as it significantly impacts the separation. However, since the separation pattern varies depending on the peptide chain lengths, amino acid compositions, and presence of modifications, optimizing separation for each peptide sequence is time-intensive. This study describes how to efficiently find the best separation conditions for peptides and related impurities utilizing LabSolutions MD, a dedicated software for supporting method development, through both screening and optimization phases.</p><h4><strong>Experimental</strong></h4><ul><li><strong>LC System: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=Nexera+X3"><strong>Nexera X3</strong></a> (Method Scouting System)</li><li><strong>MS System: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=LCMS-2050"><strong>LCMS-2050</strong></a></li></ul><h4><strong>Automated Peak Tracking by LCMS-2050&nbsp;</strong></h4><p>LC chromatograms obtained with a gradient time of 5 min, flow rate of 0.6 mL/min, and initial concentration of 5 % and 15 %, along with molecular weights for FLP and impurities, are shown in Fig. 18. The UV spectra for each impurity are displayed in Fig. 19. The similarity between UV spectra of Met(O2), p.A1del, p.A1_E2del, p.A1_K3del, and p.A1_D5del is greater than 0.99, suggesting that peak tracking based on UV spectra would be challenging. In contrast, LabSolutions MD enables peak tracking based on molecular weights acquired by LCMS-2050, facilitating accurate identification of compounds with similar UV spectra (Fig. 18). The estimated molecular weights for each compound show small mass errors compared to the theoretical values (Table 3), which can be used to confirm the molecular weights of known compounds as well as to approximate the molecular weights of unknown impurities.</p><h4><strong>Conclusion</strong></h4><p>The separation patterns of synthetic peptides vary depending on mobile phase composition, column type, and various LC parameters, such as gradient conditions, column oven temperature, and flow rate. Separation behavior can also differ based on peptide structure, including length, amino acid composition, and the presence of modifications. Therefore, it is necessary to optimize the separation for each peptide sequence individually. However, optimizing analytical conditions through numerous analyses and data processing can be time-consuming. LabSolutions MD automates the entire workflow, including generating analysis schedules, preparing mobile phases, and processing the data, thanks to functionalities such as automated peak tracking, ranking of chromatograms, and design space visualization. This article introduced a case where the optimal separation conditions for synthetic peptides were efficiently identified through screening and optimization phases. LabSolutions MD also offers fully automated gradient optimization by AI algorithm to meet user-defined criteria. For more details, please refer to the application news, “Automatic Optimization of Gradient Conditions by AI Algorithm on Synthetic Peptide and Impurities: 01-00814”.</p><h3><strong>3. Thermo Fisher Scientific / HPLC Symposium: Sensitive cationic lipids impurities analysis with quantitation by charged aerosol detection and simultaneous mass confirmation by MS</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_sensitive_cationic_lipids_impurities_analysis_quantitation_po_en_3dacf61554.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Cationic lipid is a critical component in lipid nanoparticle formulations for delivering nucleic acids such as mRNA, siRNA, antisense oligonucleotides etc. [2] Cationic lipids are amphiphilic molecules that possess a hydrophilic region, a hydrophobic region, and a linker structure connecting the two. [1] Charged aerosol detector (CAD) and evaporative light scattering detectors (ELSD) are preferred for characterizing lipid nanoparticle components, with CAD being the better choice for impurity quantification due to its higher sensitivity [3]. In a previous study, an inverse gradient method was developed to quantify impurities in cationic lipids raw materials.[1] To demonstrate the inverse gradient method on CAD HP, the same cationic lipids -- R-DOTAP, DLin-KC2- DMA and ALC-0315 -- were measured to this experiment as in Application note 003384 [1]. New diverter valve and software features were showcased, along with method’s sensitivity and repeatability.</p><h4><strong>Materials and methods</strong></h4><p>Using the inverse gradient method from Application note 003384 [1], to demonstrate impurity analysis in cationic lipids on the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/2176"><strong>Thermo Scientific Vanquish Charged Aerosol Detector HP (CAD HP)</strong></a> integrated in the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/743"><strong>Thermo Scientific Vanquish Flex Inverse Gradient UHPLC System</strong></a>, with parallel mass confirmation by the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/413"><strong>Thermo Scientific ISQ EM Single Quadrupole Mass Spectrometer</strong></a>.&nbsp;</p><p><strong>Methods:</strong> The workflow uses a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/743"><strong>Vanquish Flex Inverse Gradient LC system</strong></a> with Vanquish CAD HP and <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1970"><strong>Thermo Scientific Hypersil GOLD C8 column</strong></a>.</p><h5><strong>Data Analysis&nbsp;</strong></h5><p><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/76"><strong>Thermo Scientific Chromeleon Chromatography Data System (CDS)</strong></a> 7.2.10 was used for data acquisition and analysis.</p><h4><strong>Conclusions&nbsp;</strong></h4><p>The CAD HP integrates perfectly into the Vanquish Inverse Gradient LC system and offers high sensitivity and excellent repeatability for quantitative cationic lipid impurity analysis, while the ISQ EM MS provides straightforward mass confirmation with high repeatability, all under compliance-ready conditions.</p><h3><strong>4. Waters Corporation: Quantitative Analysis of Four Long-Chain Lysophosphatidylcholines (LPCs) in Dried Blood Spot using Liquid Chromatography Tandem Mass Spectrometry for Clinical Research</strong></h3><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/720008826_en_dc154872b6.pdf"><strong>Full PDF for download</strong></a></li></ul><h5><strong>Benefits</strong></h5><ul><li>Good chromatographic resolution of all forms</li><li>Short extraction time compared to traditional methods for lipid extraction</li><li>Short analysis injection-to-injection 7 minutes</li></ul><p>Historically, very long chain fatty acids have been quantified using direct transesterification and gas chromatography methods which involve time-consuming and labor-intensive sample preparation and analysis. Consequently, simpler LC-MS/MS methods have been reported for the measurement of long-chain LPCs (LPCs generic chemical structure, Figure 1) in DBS. Interferences have been reported using positive ESI, thus researchers have developed methodologies utilizing negative ESI to circumvent isobaric interferences when there is lack of chromatographic resolution. Here a UHPLC-MS/MS method is described using acetate adducts for the reliable analysis of four long-chain LPCs in DBS for clinical research.&nbsp;</p><p>A UHPLC-MS/MS method for quantifying a panel of LPCs (C20:0, C22:0, C24:0, and C26:0-LPCs) was developed. The mobile phase composition consisted of a mixture of water, acetonitrile, and isopropanol with 5 mM ammonium acetate as a modifier. The DBS sample extraction used a working solution prepared with the deuterated LPC internal standards in methanol. The method was developed with an ACQUITY UPLC I-Class PLUS System (Fixed Loop) using an ACQUITY Premier CSH C18 Column, which provided baseline resolution of endogenous isobaric interferences for the saturated forms of the targeted LPCs. The detector was a Xevo TQ-S micro Mass Spectrometer operated in negative ESI mode. This methodology resulted in a total analysis time (injection-to-injection) of 7 minutes.</p><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/199"><strong>ACQUITY UPLC I-Class PLUS FL&nbsp;</strong></a></li><li><strong>System Detection: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/209"><strong>Xevo TQ-S micro Mass Spectrometer&nbsp;</strong></a></li><li><strong>Plate:</strong> Waters 96-well Sample Collection Plate, 350 µL&nbsp;</li><li><strong>Column(s): </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1952"><strong>ACQUITY Premier CSH C18 Column</strong></a>, 2.1 mm x 50 mm, 1.7 µm (p/n: 186009460)</li></ul><h5><strong>MS Conditions&nbsp;</strong></h5><ul><li><strong>MS system:</strong> Xevo TQ-S micro Mass Spectrometer</li></ul><h5><strong>Data Management&nbsp;</strong></h5><ul><li><strong>MS software: </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/635"><strong>MassLynx™ v4.2 Software SCN 1045&nbsp;</strong></a></li><li><strong>Informatics: </strong>TargetLynx™ XS v4.2 Software SCN1045 and QUAN Review</li></ul><h4><strong>Results and Discussion&nbsp;</strong></h4><p>During method development, different types of column chemistry, mobile phase composition, and ESI polarity modes were assessed to optimize chromatographic peak shape, analytical sensitivity and manage interferences. In positive mode, ammonium formate favored the formation of m/z 184 and m/z 104 as daughter ions for all the LPCs. The m/z 184 ion lacks specificity because most phospholipids share this moiety, so the ion m/z 104 was selected as quantifier. Having evaluated conditions in positive mode for the analysis of the four saturated LPCs, it was observed that some existing interferences were not only affecting the parent masses of the analytes but their corresponding internal standard parent ion masses too. Hence, negative ionization using ammonium acetate as modifier was assessed for the detection of LPCs. Acetate adducts [M+CH3COO]- were the most abundant parent ion masses, generated under these conditions for each LPC. The acyl-chain product ions generated in negative ionization reduce the detection of interferences coming from other phosphocholine headgroups. The latter detection conditions were used for the validation of the method presented herein.</p><h4><strong>Conclusion&nbsp;</strong></h4><p>An LC-MS/MS clinical research method for measuring C20:0, C22:0, C24:0, and C26:0 LPCs with the ACQUITY UPLC I-Class PLUS FL System, and the Xevo TQ-S micro Mass Spectrometer that:</p><ul><li>Demonstrates good linearity, with no significant carryover, or matrix effects</li><li>Offers good precision with a total reproducibility and repeatability <span>≤</span>10% CV</li><li>Offers reliability in the identification of LPCs from interfering isobaric compounds Provides good selectivity and analytical sensitivity in negative mode and is relatively fast as approximately 220 samples can be analyzed in 24 hours</li></ul>

News from LabRulezLCMS Library - Week 39, 2025

<p><strong>Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 29th 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 note by <strong>Agilent Technologies </strong>and posters by<strong> Shimadzu / AOAC, Thermo Fisher Scientific </strong>and <strong>Waters Corporation / AOAC!</strong></p><h3><strong>1. Agilent Technologies: Comparison of Dual Sorbent Solid Phase Extraction for PFAS Applications</strong></h3><p><strong>Using dual-phase Agilent </strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=Bond+Elut+PFAS+WAX"><strong>Bond Elut PFAS WAX/ Carbon S SPE</strong></a><strong> cartridges for US EPA Method 1633</strong></p><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/an_1633_wax_carbon_s_spe_comparison_5994_8291en_agilent_8bdc559ad8.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Many laboratories have opted to combine polymeric weak anion exchange (WAX) and graphitized carbon black (GCB) into single solid phase extraction (SPE) cartridges to streamline sample preparation and facilitate automation for the determination of Per- and Polyfluoroalkyl Substances (PFAS) in environmental samples. This modification is acceptable under Section 1.5 of US EPA Method 1633, if the method's quality control metrics are met.1 The Agilent PFAS WAX and Carbon&nbsp;S sorbents are specifically manufactured for PFAS applications and are lot-tested to ensure low PFAS residue and maximum recovery.2,3 Packing these sorbents into standard 6 mL SPE cartridges with low PFAS residue frits makes them ideally suited for automated methods requiring both WAX extraction and carbon matrix reduction. The extraction performance of the dual-phase Agilent Bond Elut PFAS WAX/Carbon&nbsp;S SPE cartridges are comparable to other commercial cartridges. This ensures seamless integration into existing workflows while assuring optimal performance for PFAS applications.</p><h4><strong>Experimental&nbsp;</strong></h4><p>Extraction recoveries using the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=Bond+Elut+PFAS+WAX"><strong>Bond Elut layered 200 mg PFAS WAX (top)/50 mg Carbon S (bottom) SPE cartridges</strong></a> (part number 5610-2237) were compared to two other commercial 200 mg WAX/50 mg GCB dual-phase cartridges (Table 1). Four replicate extractions were performed with each sorbent, except for benchmark B, where only two replicates were extracted. The target compounds included the 40 listed in Method 1633, spiked at a low-level concentration of 4 ng/L in 250 mL of reagent water. Target recoveries were quantified using 24 isotope-labeled extracted internal standards (EIS). Extractions were carried out using automated SPE. Method details were described in a previous application note.4</p><h4><strong>Conclusion&nbsp;</strong></h4><p>Dual-phase Agilent Bond Elut PFAS WAX/Carbon S SPE cartridges are specifically manufactured for PFAS applications, with rigorous lot testing to ensure PFAS cleanliness and recovery. They enable efficient streamlined sample preparation and automation for EPA Method 1633. PFAS target and extracted internal standards recoveries are comparable to other dual-phase SPE cartridges, facilitating easy integration into existing EPA Method 1633 workflows.</p><h3><strong>2. Shimadzu / AOAC: Analysis of Per- and Polyfluoroalkyl Substances (PFAS) in Drinking Water and Milk using LC-MS/MS coupled with Online-SPE Interface</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/aoac_2025_t009_835132523a.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Per- and Polyfluoroalkyl Substances (PFAS) are a chemical group of organic fluorinated compounds, and perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) are representative compounds. They have been used in applications such as water repellents, surface treatment agents, fire extinguishers, and coatings. PFAS are known to be widely present in the environment and wildlife due to their stable structure. Concerns about human exposure through diet have led various countries to conduct investigations into food contamination by PFAS.</p><p>We have examined a quantitative analysis method for 43 PFAS in tap water, drinking water, and milk, and developed a pretreatment and analysis method that realized labor and time savings using LC-MS/MS and automatic pretreatment equipment (Fig.1). For extraction from milk, the QuEChERS method was used with reference to the FDA method.</p><h4><strong>Experimental</strong></h4><ul><li><strong>Automatic pretreatment equipment: </strong>SPL-W100 (AiSTI SCIENCE)</li><li><strong>LC-MS/MS:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=LCMS-8060RX"><strong>LCMS-8060RX</strong></a> (Shimadzu)</li></ul><h4><strong>Conclusion</strong></h4><ul><li>LC-MS/MS methods were developed to analyze 43 PFAS in 11 minutes using automatic pretreatment, saving time and labor.</li><li>We conducted recovery tests by adding samples to tap water, drinking water, and milk, and achieved the LOQ set by EPA regulations and AOAC criteria for all compounds.</li><li>The quantitative values of the four main components in tap water were PFOA 0.00109 <span>μ</span>g/kg, PFNA 0.00044 <span>μ</span>g/kg, PFHxS 0.0004 <span>μ</span>g/kg and PFOS 0.00063 <span>μ</span>g/kg. PFAS was not detected above the detection limit in drinking water and milk.</li></ul><h3><strong>3. Thermo Fisher Scientific: High-Throughput and Highly Selective Quantitative Lipidomics with the Stellar Mass Spectrometer – A Novel Hybrid Nominal Mass Instrument</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_unil_stellar_lipidomics_metsoci_2025_po_en_730b1c19fd.pdf"><strong>Full PDF for download</strong></a></li></ul><p>The increasing prevalence of metabolic disorders underscores the need for robust, high-throughput clinical lipid panels and lipidomics methods to quantify lipid species comprehensively, with high precision and selectivity . Structurally related lipids, with varying fatty acid composition, were associated with distinct metabolic consequences and proven to have different biological roles. Thereby, measuring individual lipid species with highest selectivity possible is crucial for corroboration of clinical utility including the improved risk prediction and diagnostics. We present an advanced methodology leveraging the unique capabilities of the new Stellar mass spectrometer. This nominal mass instrument addresses current limitations in specificity and throughput by enabling fast acquisition parallel reaction monitoring, and revolutionizing lipidomics workflows by delivering unprecedented specificity and quantitation. This system combines a quadrupole and a dual cell linear ion trap that significantly improves the ion accumulation efficiency and scan speed. This allows for the acquisition of full MS2 spectra including multiple transitions, thus improving the annotation specificity by resolving isomeric lipids and structurally similar species. This unique capacity of Stellar MS positions it as a practical solution for translational research, biomarker discovery and clinical lipidomics where rapid, sensitive and selective lipid quantitation is essential.</p><h4><strong>Materials and Methods&nbsp;</strong></h4><h5><strong>Sample Preparation&nbsp;</strong></h5><p>Dried UltimateSPLASH ONE internal standard mixture (100 µL) was reconstituted in 25 µL of NIST human plasma SRM 1950 followed by the addition of 125 µL of isopropanol for lipid extraction. The supernatant was then transferred to LC vials for LC-MS analysis. Linearity assessment: different volumes of Avanti Research UltimateSPLASH ONE internal standard mixture (0.5, 1, 5, 10, 50, 100, and 500 µL) were evaporated to dryness, followed by the reconstitution with 25 µL NIST Plasma and 125 µL of isopropanol for lipid extraction.&nbsp;</p><h5><strong>Data Acquisition&nbsp;</strong></h5><p>Plasma lipid extracts were separated on a ACQUITY Premier BEH Amide column (1.7um, 100 mm x 2.1 mm I.D., Waters) and analyzed using a targeted LCMS lipidomics approach on <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/742"><strong>Thermo Scientific Vanquish Duo UHPLC system</strong></a> coupled to a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1149"><strong>Stellar mass spectrometer</strong></a> (Figure 1) 1 . This method was built from a well-established selected reaction monitoring (SRM) method that we Converted to a parallel reaction monitoring (PRM) method by using only the unique precursors. As there are many isomeric lipids sharing the same mass, converting the method for the Stellar reduced 1,019 (579 positive mode; 440 negative mode) SRM precursors to 524 (233 positive mode; 291 negative mode) PRM precursors.</p><h4><strong>Conclusions&nbsp;</strong></h4><ul><li>By integrating LipidCreator with Stellar MS, we expanded on an already thorough lipid workflow by increasing the number of targeted lipids (&gt;50%) and transitions (&gt;3/molecule) enabling higher confidence in lipid identification&nbsp;</li><li>Multiple transitions per precursor, combined with fast scan speeds, enhanced both the specificity and accuracy of lipid quantification, particularly for isomeric species.&nbsp;</li><li>Employing both HCD and CID simultaneously allowed for more robust characterization of sphingomyelins seen in the 2-3 transitions per species.&nbsp;</li><li>This high throughput, targeted approach addresses key limitations and provides a efficient platform for biomarker discovery&nbsp;</li><li>Also, look for our other poster showing a high-throughput targeted MS3 approach has been optimized on the Stellar MS, for the semi-quantification of TG species in human plasma.</li></ul><h3><strong>4. Waters Corporation / AOAC: From Coop to Carton: A Study of PFAS in Backyard &amp; Store-Bought Eggs Using Automation and LC-MS/MS</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/720009037_6848187630.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Consumers often view backyard chicken eggs as a healthier alternative to store-bought eggs. However, environmental factors such as contaminated soil, water, feed, and bedding may expose backyard chickens to per- and polyfluoroalkyl substances (PFAS). This study set out to compare PFAS levels in eggs from backyard flocks with those in cage-free eggs purchased from grocery stores. The challenge was to analyze whole egg—a dense protein- and fat-rich matrix—efficiently and reproducibly. To address this, an automated sample preparation workflow was developed, reducing analyst involvement, minimizing variability, and ensuring robust results.</p><h4><strong>Methods</strong></h4><p>The workflow combined automated extraction (CEM EDGE) with automated solid-phase extraction (PromoChrom SPE-03) using dual-phase <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/215"><strong>Oasis GCB/WAX cartridges</strong></a> designed for PFAS. Key steps included:</p><ul><li>Homogenizing whole eggs and spiking with internal standards.</li><li>Automated alkaline extraction cycles followed by nitrogen-assisted concentration.</li><li>SPE cleanup with GCB/WAX cartridges to remove matrix interferences.</li><li>Analysis by LC-MS/MS using a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/191"><strong>Waters ACQUITY Premier system</strong></a> coupled to a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/554"><strong>Xevo TQ Absolute mass spectrometer</strong></a>.</li></ul><p>This approach enabled preparation in less than 15 minutes per sample, while the SPE system processed up to eight samples simultaneously in under 70 minutes. Altogether, the workflow provided consistent recoveries across a broad range of PFAS, including EU-regulated compounds such as PFOA, PFOS, PFNA, and PFHxS.</p><h4><strong>Conclusions</strong></h4><p>Automated sample preparation and SPE enabled reproducible, high-throughput analysis of PFAS in whole eggs—a challenging matrix. Results showed that backyard eggs generally contained higher PFAS concentrations and a wider variety of compounds than store-bought eggs, reflecting greater environmental exposure. In contrast, grocery store eggs contained certain PFAS precursors, suggesting possible legacy contamination. The method proved robust for both regulated and emerging PFAS, with recoveries in the acceptable 70–130% range. This workflow offers laboratories a reliable, future-proof solution for PFAS monitoring in foods, helping to ensure consumer safety and regulatory compliance.</p>

News from LabRulezLCMS Library - Week 40, 2025

<p><strong>Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 6th October 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 other document by <strong>Agilent Technologies </strong>and posters by<strong> Shimadzu / AOAC, Thermo Fisher Scientific / ASMS </strong>and <strong>Waters Corporation / AOAC!</strong></p><h3><strong>1. Agilent Technologies: PFAS Food Legislation Overview</strong></h3><ul><li>Other document</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/ebook_pfas_food_legislation_5994_8575en_agilent_dfe5a6222c.pdf"><strong>Full PDF for download</strong></a></li></ul><p>This <strong>Agilent Technologies eBook</strong> provides an overview of <strong>global PFAS (per- and polyfluoroalkyl substances) regulations and analytical methods</strong> for monitoring these compounds in food, beverages, and food-contact materials. PFAS are synthetic chemicals valued for their water- and grease-resistant properties but are highly persistent and bioaccumulative. Their potential health risks—including immune suppression, elevated cholesterol, liver enzyme changes, and developmental effects—have prompted worldwide regulatory action to limit exposure through food and packaging.</p><p>In the <strong>European Union</strong>, <strong>Regulation (EU) 2023/915</strong> establishes maximum allowable levels of PFAS in various food products (e.g., meat, fish, eggs), ranging from 0.2 to 8 µg/kg depending on the matrix. <strong>Recommendation (EU) 2022/1431</strong> encourages member states to collect PFAS occurrence data and harmonize testing methods, while the upcoming <strong>Packaging and Packaging Waste Regulation (PPWR 2025/40)</strong> will, from August 2026, limit PFAS in food packaging to <strong>25 ppb per compound</strong> and <strong>250 ppb total</strong>. Analytical testing across Europe relies primarily on <strong>liquid chromatography coupled with tandem mass spectrometry (</strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?instrumentationItems=32&amp;mainauthorItems=10"><strong>LC/MS/MS</strong></a><strong>)</strong>, which provides the ppb-to-ppt sensitivity needed for regulatory compliance.</p><p>In the <strong>United States</strong>, the framework combines federal, state, and industry standards. The <strong>USDA CLG-PFAS 2.04</strong> method employs <strong>UHPLC/MS/MS</strong> to detect 16 PFAS in animal tissues, while the <strong>FDA C-010.03</strong> method quantifies 30 PFAS in food and feed using <strong>solid-phase extraction (SPE)</strong> followed by <strong>LC/MS/MS</strong>. The <strong>AOAC SMPR 2023.003</strong> standard defines performance criteria for PFAS testing across multiple food categories, ensuring validated and reproducible results. Several U.S. states—including Maine, California, and Washington—have also enacted specific limits for PFAS in meat, milk, and food-contact packaging.</p><p>In <strong>China</strong>, national standard <strong>GB 5009.253-2016</strong> specifies the determination of PFOS and PFOA in animal-derived foods via <strong>isotope-dilution LC/MS/MS</strong>. A new draft revision expands testing to 30 PFAS compounds (including isomers) across broader food matrices such as fish, vegetables, and milk. Chinese authorities have also introduced bans and restrictions on PFOS, PFOA, and PFHxS production and use, along with environmental management and disposal requirements.</p><p>Agilent offers <strong>comprehensive PFAS testing solutions</strong> for food laboratories, including robust <strong>HPLC and LC/MS instrumentation</strong>, validated methodologies aligned with EPA, EU, and ASTM standards, and a curated <strong>PFAS MRM database</strong>. The portfolio also features <strong>Captiva EMR PFAS Food cartridges</strong> pre-tested for background contamination, certified PFAS standards, and data-handling tools such as <strong>Agilent MassHunter</strong> and <strong>SLIMS</strong>. Together, these workflows enable reliable PFAS quantification, reduced contamination risk, and full compliance with international food-safety legislation.</p><h3><strong>2. Shimadzu / AOAC: Reliable and direct quantitative analysis of multi-mycotoxins in animal feed by using Shimadzu LCMS-8050RX</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/aoac_2025_m040_3bceabefcd.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Mycotoxins are toxic compounds produced by moulds that contaminate crops such as cereals, nuts, spices, dried fruits, apples, and coffee especially under warm, humid conditions. These chemically stable toxins can persist through food processing and pose serious health risks to humans and animals, ranging from acute poisoning to cancer and immunosuppressive. Among the many identified toxic compounds, Aflatoxins, Ochratoxin A, Fumonisins, T-2, Zearalenone, and Deoxynivalenol (DON) (Figure 1) are of particular concern. These are the most studied and regulated due to their significant impact on human and animal health, making their testing critically important. While conventional testing methods (HPLC-UV/FLD) are still common, regulatory focus is shifting toward LC-MS/MS testing due to its sensitivity and specificity.</p><p>The European Committee for Standardization (CEN), under the M/520 mandate, is developing standardized methods for this purpose. Mycotoxins are generally extracted via liquid–liquid (for milk, wine, juices) or solid–liquid extraction (for cereals, dried fruits, spices, feed), typically using acidic buffers. In this study, we developed a fast, highthroughput LC-MS/MS method for multi-mycotoxin detection in animal feed using the Shimadzu LCMS8050RX (Figure 2). MRM based quantitative method was developed using Shim-pack GIST C18-AQ HPLC column with an aim to achieve improved peak separation and required sensitivities for the compounds under study. A matrix-matched calibration curve (0.5–50.0 ppb) showed accuracy between 80–120 %, and recoveries (at 2.5 and 5.0 ppb) were 70–120 %, complying with SANTE guidelines[1] .</p><h4><strong>Materials and method</strong></h4><h5><strong>LC-MS/MS analysis</strong></h5><ul><li><strong>UHPLC:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=Nexera+X3"><strong>Nexera X3</strong></a></li><li><strong>MS:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=LCMS-8050RX"><strong>LCMS-8050RX</strong></a></li></ul><h4><strong>Conclusion&nbsp;</strong></h4><ul><li>Average recovery values for multi-mycotoxins in animal feed sample were found to be within the acceptance criteria as per the SANTE/12682/2019 guidelines.&nbsp;</li><li>The results obtained at 2.5 ppb and 5.0 ppb concentration level were accurate, repeatable and reproducible with RSD less than 20 %.</li><li>A simple liquid-liquid extraction method has been successfully developed and validated for the simultaneous quantification of nine mycotoxins in a single run.</li></ul><h3><strong>3. Thermo Fisher Scientific / ASMS: High-throughput proteomics using narrow window DIA on the Orbitrap Astral Zoom MS</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_407_narrow_window_dia_proteomics_po407_asms2025_en_eea95fbc58.pdf"><strong>Full PDF for download</strong></a></li></ul><p>The recently introduced Orbitrap Astral mass spectrometer has significantly expanded the scale and scope of proteomics experiments, advancing discovery and translational research. Building on this innovation, we demonstrate the performance of the next evolution of the platform, the Orbitrap Astral Zoom mass spectrometer, at throughputs ranging from 30 to 300 samples-per-day (SPD) analysing a HeLa digest using narrow window data-independent (nDIA) acquisition. The novel instrument features a higher acquisition rate (270 Hz vs. 200 Hz) achieved through improved ion optics settling times and faster ion transfers, increased sensitivity via pre-accumulation, and deeper coverage through enhanced spectral processing, among other improvements. Comparative analysis with the Orbitrap Astral MS reveals further improvements in peptide and protein group identifications enabling even deeper coverage at high throughput whilst maintaining precise quantitation.</p><h4><strong>Methods</strong></h4><p>HeLa digest standard measured on <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1959"><strong>Orbitrap Astral Zoom MS</strong></a> and Thermo Scientific <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/836"><strong>Orbitrap Astral MS</strong></a> coupled to Thermo Scientific <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/398"><strong>Vanquish Neo UHPLC</strong></a> utilizing 30-300 samples-per-day (SPD) LC-MS methods</p><h4><strong>Conclusions&nbsp;</strong></h4><p>The novel Orbitrap Astral Zoom mass spectrometer is, among its broad and comprehensive range of applications, an excellent instrument for high-throughput proteomics.&nbsp;</p><ul><li>High-throughput nDIA on the Orbitrap Astral Zoom MS generates deep coverage of proteomes</li><li><span>&nbsp;</span>The novel Orbitrap Astral Zoom MS further pushes the boundaries in terms of quantity and quality of identifications&nbsp;</li><li>The Orbitrap Astral Zoom MS enables precise quantitation with full flexibility to focus on MS2-based quantitation</li></ul><h3><strong>4. Waters Corporation / AOAC: Integration of Ultra-short Chain PFAS Into Routine Analysis Methods: Addressing Retention and Confirmatory Ions</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/720009019_f2e0c3e56f.pdf"><strong>Full PDF for download</strong></a></li></ul><p>The poster presents a novel LC-MS/MS workflow enabling simultaneous quantification and confirmation of ultra-short through long-chain PFAS in a single analysis. The work was conducted using a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/191"><strong>Waters ACQUITY Premier LC system</strong></a> equipped with a <strong>PFAS kit</strong>, <strong>Atlantis Premier BEH C18 AX mixed-mode column</strong>, and a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/554"><strong>Xevo TQ Absolute tandem mass spectrometer</strong></a> with <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1544"><strong>waters_connect software</strong></a> for quantitation.</p><p>The study addresses two analytical challenges: (1) the poor chromatographic retention of ultra-short chain PFAS (e.g., TFA, PFPrA) on conventional reverse-phase columns and (2) the absence of confirmatory ions for some short-chain PFAS, which typically produce limited fragment ions. Using the <strong>Atlantis Premier BEH C18 AX column</strong>, which combines reversed-phase and anion-exchange mechanisms, the authors achieved stable retention of C2–C4 PFAS. Optimized pH control using ammonium hydroxide and acetic acid further stabilized retention times of highly polar compounds.</p><p>For mass spectrometric confirmation, the team demonstrated the use of the <strong>m/z 19 fragment transition</strong> as a new confirmatory ion for PFAS that previously lacked one. On the <strong>Xevo TQ Absolute system</strong>, low-mass transmission enhancement significantly increased sensitivity for this ion, allowing confirmatory detection of PFBA and PFPeA even at low (ppt-level) concentrations. Ion ratio stability across calibration curves met common method validation criteria (±30%), demonstrating robustness for routine use.</p><p>The combined method enables the <strong>simultaneous analysis of ultra-short to long-chain PFAS</strong> in a single injection without requiring additional columns or HRMS verification. Results from landfill leachate and surface water samples confirmed the method’s capability to resolve co-eluting interferences and to reliably identify PFAS at environmentally relevant levels. This workflow offers an efficient and practical solution for laboratories transitioning to include ultra-short chain PFAS within standard monitoring methods.</p>

News from LabRulezLCMS Library - Week 41, 2025

<p><strong>Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 13th October 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,</strong> <strong>Shimadzu </strong>and<strong> Waters Corporation </strong>and poster by<strong> Thermo Fisher Scientific / HPLC!</strong></p><h3><strong>1. Agilent Technologies: Expanding Coverage for the Analysis of PFAS in Paper-Based Food Packaging Materials</strong></h3><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/an_pfas_food_contact_materials_6545_lcqtof_5994_8516en_agilent_0d3cbb1f4d.pdf"><strong>Full PDF for download</strong></a></li></ul><p>PFAS are manufactured chemicals used in food packaging for their grease- and waterproof properties. However, PFAS are persistent in the environment and may pose health risks.2 This study presents a workflow to screen for PFAS in FCMs using LC/Q-TOF. The FluoroMatch software suite was used to automate PFAS data annotation of knowns and unknowns as well as generate an interactive visualization dashboard, enhancing analysis accuracy and coverage.&nbsp;</p><p>The FluoroMatch suite is an open-source set of tools designed to streamline the suspect and nontarget screening of PFAS compounds. It automates several processes, including file conversion, chromatographic peak picking, blank feature filtering, PFAS annotation based on precursor and fragment masses, homologous series detection, compound classification (PFAS versus not PFAS), and reporting annotation confidence. The software library contains 15,643&nbsp;PFAS species and associated fragmentation patterns, with the capability to add more.3&nbsp;</p><p>Various FCM samples, including takeaway containers and microwave popcorn bags, were rigorously extracted and analyzed. Suspect screening was employed for PFAS quantitation, while nontargeted workflows enabled the identification and semi-quantitation of PFAS that were lacking&nbsp;standards.</p><h4><strong>Experimental</strong></h4><h5><strong>LC/MS analysis&nbsp;</strong></h5><p>Extracts were analyzed using an <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=1290+Infinity+II+LC"><strong>Agilent 1290 Infinity II LC system</strong></a> coupled with an Agilent <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=6545+LC%2FQ-TOF"><strong>6545 LC/Q-TOF</strong></a>. An Agilent <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=PFC-Free+HPLC+Conversion+kit"><strong>PFC-Free HPLC Conversion kit</strong></a> (part number 5004‑0006) and an Agilent <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=InfinityLab+PFC+delay+column"><strong>InfinityLab PFC delay column</strong></a> (part number 5062‑8100) were used to minimize background PFAS levels. Liquid chromatographic separation was performed on a 1.8&nbsp;µm, 2.1 × 100 mm, Agilent ZORBAX RRHD StableBond SB‑C18 column (part number 858700-902) with a corresponding guard column (Agilent part number 821725-902). The mobile phases were (A) water with 20&nbsp;mM ammonium acetate and (B) methanol. The flow rate was 0.4&nbsp;mL/min. The elution gradient was: 5% B (0 to 0.5&nbsp;minutes), linear increase to 40% B (0.5 to 3 minutes), linear increase to 80% B (3 to 16 minutes), hold 80% B (16 to 18 minutes), linear increase to 100% B (18 to 22 minutes), hold 100% B (20 to 22&nbsp;minutes), decrease to 5% B (22 to 22.5 minutes), followed by a 4 minutes post‑run re‑equilibration. The injection volume was 10 μL, and the column temperature was maintained at 50 °C. For the LC/Q‑TOF, the drying gas was set to 4 L/min at 230 °C, the nebulizer to 20 psi, and the sheath gas to 12 L/min at 375 °C.</p><h4><strong>Results and discussion</strong></h4><p>PFAS identification using <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=FluoroMatch"><strong>FluoroMatch</strong></a> Using FluoroMatch Visualizer, interactive tools for visualizing PFAS data were generated, including mass defect plots, accurate mass versus retention time plots, MS/MS fragmentation plots, annotation tables, and fragment screening. Individual homologous series were selected based on nominal mass and normalized mass defect, allowing for the observation of patterns and identification of outliers. This interactive cross-filtering simplified the evaluation of PFAS features and enhanced confidence in nontargeted results.&nbsp;</p><p>FluoroMatch was used to annotate a homologous series (C3 to C14) of perfluoroalkyl carboxylic acids (PFCAs), including PFHxA (perfluorohexanoic acid), PFHpA (perfluoroheptanoic acid), PFOA (perfluorooctanoic acid), PFNA (perfluorononanoic acid), PFDA (perfluorodecanoic acid), and PFDoDA (perfluorododecanoic acid). This workflow demonstrated that the suspect screening approach successfully identified common PFAS with available standards, while FluoroMatch was able to annotate additional PFAS that were lacking standards.&nbsp;</p><p>Additionally, FluoroMatch annotated C6 and C8 perfluoroalkyl sulfonic acids (PFSAs), which were not identified by suspect screening. This highlights the value of a hybrid workflow that combines traditional suspect screening with nontargeted tools like FluoroMatch, enhancing confidence in the nontargeted results through their complementary nature. Specifically, FluoroMatch allows for homologous series detection to help annotate incomplete or noisy spectra. FluoroMatch Visualizer evaluation helps incorporate additional lines of evidence to annotation like retention time patterns and Kendrick mass defect to aid in annotation.</p><h4><strong>Conclusion&nbsp;</strong></h4><p>This study successfully demonstrates the effectiveness of FluoroMatch software in automating the annotation and visualization of PFAS compounds in food packaging materials. The integration of FluoroMatch Visualizer provides a comprehensive approach to identifying both targeted and nontargeted PFAS compounds. The results highlight the prevalence of PFAS in various paper‑based FCMs, with clamshell to-go boxes showing the highest concentrations, particularly PFOA and PFDA, which exceed European Union regulatory limits. As PFAS can migrate into food, especially at higher temperatures, the pervasiveness of PFAS in FCMs raises concerns about potential exposure through hot meals and microwave heating. The widespread detection of PFAS in clamshell to-go boxes and other FCMs indicates a need for stricter regulations to reduce PFAS use in food packaging to protect consumer&nbsp;health.</p><h3><strong>2. Shimadzu: An Ultra-high Sensitivity Analysis of 29 PFAS Compounds in Drinking Water by Direct Injection</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_01026_en_dcf554dd6f.pdf"><strong>Full PDF for download</strong></a></li></ul><p>PFAS (Per- and Polyfluoroalkyl Substances) are synthetic fluorinated organic compounds. More than a few thousand compounds exist, each with a varied carbon chain length, functional groups, and structural isomers. Because of its useful qualities, such as their ability to repel water and their nonstickiness, PFAS are found in a wide range of consumer and industrial products, including firefighting foams and coatings. Their high stability and persistence raise concerns about their impact on human health and the environment. In recent years, regulations and studies regarding PFAS have been increasing globally. For example, the U.S. Environmental Protection Agency (EPA) has set a Maximum Contaminant Level (MCL) of 4 ng/L for PFOA and PFOS in drinking water. To improve the analysis of PFAS, more sensitive, accurate and faster methods are needed. This application introduces the ultra-high sensitivity analysis of 29 PFAS in drinking water by direct injection using the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=LCMS-8065XE"><strong>LCMS-8065XE</strong></a> (Fig. 1) and addresses the demands of testing laboratories for PFAS quantification.</p><h4><strong>Analytical conditions</strong></h4><p>The HPLC and MS analytical conditions are shown in Table 1 and 2. The test compounds used were the 29 compounds included in methods EPA 533 and 537.1 for drinking water analysis. The <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=LCMS-8065XE"><strong>LCMS8065XE</strong></a> is equipped with a new ESI probe (StreamFocus) and a new collision cell (UFsweeper IV), providing more sensitive and accurate results in PFAS analysis. A delay column was installed between the mixer and autosampler to separate PFAS from the HPLC system and those in the sample. To ease the analysis, same type of column is used as delay and analytical columns.</p><h4><strong>Conclusion&nbsp;</strong></h4><ul><li>29 of PFAS in drinking water were measured with high sensitivity (&lt;1 ng/L) by direct injection method using the LCMS-8065XE.&nbsp;</li><li>Good recovery and reproducibility were obtained in drinking water samples, confirming that this analytical method can deliver accurate quantification for PFASsin drinking water.</li></ul><h3><strong>3. Thermo Fisher Scientific / HPLC: Efficient tandem capillary flow LC-MS with short μPAC columns and a single ionization source</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_efficient_tandem_capillary_flow_lc_ms_short_pac_columns_po_en_f38e078c9a.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Nano-flow tandem direct injection (TDI) workflows increase sample throughput and instrument productivity with flow rates below 1 µL/min but need dual-column ESI interfacing to maintain chromatographic performance. At flow rates above 1 µL/min with capillary flow columns, post-column dispersion impacts performance less, allowing low-dispersion switching valves in the fluidic path. We demonstrate the Vanquish Neo UHPLC system TDI workflow for capillary flow LC with a high throughput short pillar array column. This dual-column, single-ESI emitter setup reduces variability, extends emitter life through efficient salt management, and ensures high reproducibility with dedicated gradient pumps and intelligent method design.</p><h4><strong>Materials and methods</strong></h4><h5><strong>Sample Preparation&nbsp;</strong></h5><p>HeLa cell digests were resuspended in 0.1% TFA and 1% ACN to obtain a 200 ng/µL stock solution, sonicated, diluted, and vortexed before use.</p><h5><strong>LC-MS configuration&nbsp;</strong></h5><p>Samples were analyzed using a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/398"><strong>Vanquish Neo UHPLC</strong></a> instrument that was configured in tandem direct injection mode with an additional pump module and a 10 µL injection loop for maximum throughput. Two µPACM Neo High-Throughput Plus columns were positioned between low-dispersion 6-port switching valves in the column compartment. A 20 µm ID nanoViper line connected the alternating eluents to an ESI emitter with an integrated liquid junction, positioned in a Thermo Scientific EASY-Spray source coupled to a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/402"><strong>Orbitrap Exploris 240 mass spectrometer</strong></a></p><h4><strong>Conclusions&nbsp;</strong></h4><ul><li>Using optimized µPAC Neo HT Plus columns in combination with the Tandem Vanquish Neo UHPLC system heightened the productivity of bottom-up proteomics measurements to 86 - 92% (gradient-dependent)&nbsp;</li><li>At these throughputs, choice of emitter only has a minor effect on identifications, FWHM and TIC. Decreasing the flow rate to 1 uL/min causes peak broadening and increases ionization efficiency which is reflected in higher TICs. In this study, reducing the flow rate only showed a positive effect on protein IDs for the longest gradient (100 SPD).&nbsp;</li><li>With the optimized methods, identifications up to 4713 protein groups for the 180 SPD and 5692 protein groups for the 100 SPD were achieved. The CVs using the same column were between 5-6% and between two columns at around 7-8% (n=6)&nbsp;</li><li>Column reproducibility was showcased for 12 columns across four different production batches. Overall, the ID rate variation between the columns was 1.4%. The median coefficient of variation of the retention times of spiked-in PRTC peptides across the tested 12 columns was 1.8%</li></ul><h3><strong>4. Waters Corporation: Oligo Mapping of sgRNA Digests: Leveraging Xevo MRT Mass Spectrometer Performance and Streamlining Data Analysis</strong></h3><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/720009009_en_f35f22bebc.pdf"><strong>Full PDF for download</strong></a></li></ul><h5><strong>Benefits&nbsp;</strong></h5><ul><li>Confident characterization of sgRNA digestion products with sub-ppm mass accuracy of oligonucleotide precursors, outstanding MS resolution (~100,000), and sensitivity at high acquisition speed</li><li>A compliance-ready informatics workflow featuring the waters_connect MAP Sequence App (ver 1.0) streamlines oligonucleotide mapping of sgRNA digests</li><li>RapiZyme MC1, a new ribonuclease - offers unique cleavage specificity and the opportunity to generate overlapping digestion products, enabling complete (100%) sequence coverage for sgRNAs</li></ul><p>Single guide RNAs (sgRNAs) were first described in 20121 when two RNA molecules essential for CRISPR-Cas9- mediated DNA cleavage – the trans-activating CRISPR RNA (tracrRNA), which serves as a scaffold for the Cas9 nuclease, and the CRISPR RNA (crRNA), responsible for DNA target recognition - were fused into a single RNA construct. This fusion created the sgRNA, a critical component of the CRISPR-Cas9 gene editing system.<sup>1–2</sup> The sgRNA molecule directs the Cas9 nuclease to introduce precise double-stranded breaks in DNA, enabling targeted genetic modifications.&nbsp;</p><p>Since its discovery, which was honored with the Nobel Prize in Chemistry, CRISPR technology has transformed gene editing applications. Beyond basic research, it has been adapted for rapid diagnostic tools, such as COVID19 tests developed during the 2020 global pandemic<sup>3</sup> , and holds promise for therapeutic interventions in genetic diseases, cancer, and infectious diseases. As sgRNAs are typically synthesized via solid-phase oligonucleotide synthesis, their analytical characterization requires confirmation of both the intact molecular weight<sup>4</sup> and the sequence verification to ensure functional accuracy.<sup>5</sup>&nbsp;</p><p>Traditional LC-MS workflows for sgRNA digest oligo mapping have been labor-intensive and time-consuming, relying heavily on manual data analysis and interpretation. Recently<sup>6</sup> , a UPLC-MS and informatics workflow designed for automated sgRNA sequence mapping was presented following enzymatic digestion with a panel of ribonucleases, including RNase T1, hRNAse 4 and two RNase T2 enzymes: RapiZyme MC1 and Cusativin. This application note extends the effectiveness of this workflow (Figure 1) when implemented on a Xevo MRT Mass Spectrometer (Figure 1), while focusing specifically on MC1 digestion of the Waters™ sgRNA LC-MS standard, where complete unique digested product coverage was obtained.</p><h4><strong>Experimental</strong></h4><h5><strong>LC Conditions</strong></h5><ul><li><strong>LC-MS system:</strong> <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1148"><strong>Xevo MRT</strong></a> (multi-reflecting time-of-flight) Mass Spectrometer coupled with <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/191"><strong>ACQUITY™ Premier UPLC (Binary) System&nbsp;</strong></a></li><li><strong>Column: </strong>ACQUITY Premier Oligonucleotide BEH™ C18 Column 300 Å, 1.7 µm, 2.1 x 150 mm, (p/n:186010541)</li></ul><h4>Results and Discussion&nbsp;</h4><p>An informatics workflow (Figure 1) featuring the waters_connect MAP Sequence App (version 1.0) was used to facilitate automated data processing of UPLC-MSE datasets acquired following the enzymatic digestion of an sgRNA standard with RapiZyme MC1.6–7 The informatics processing workflow consisted of three steps:&nbsp;</p><ol><li><strong>In-silico digestion:</strong> The mRNA Cleaver MicroApp was used to generate predicted, in-silico digested oligonucleotide digestion products, based on the target RNA sequence.&nbsp;</li><li><strong>Data processing and mass matching:</strong> The waters_connect MAP Sequence App processed the UPLC-MSE data, matching the predicted neutral monoisotopic masses of digested oligonucleotides to the experimentally acquired MS1 data.&nbsp;</li><li><strong>Sequence coverage visualization: </strong>The resulting sequence coverage for the RNA digest was summarized and visualized using the Coverage Viewer MicroApp.</li></ol><h4><strong>Conclusion&nbsp;</strong></h4><ul><li>An informatics workflow, incorporating the waters_connect MAP Sequence App (ver 1.0) was successfully demonstrated for UPLC-MSE digested oligo mapping analysis of sgRNA digests using the Xevo MRT QTOF Mass Spectrometer.&nbsp;</li><li>Confident peak assessments for analysis of sgRNA digests were achieved with the Xevo MRT QTOF Mass Spectrometer, which delivered excellent sensitivity, high resolution MS (100,000), and mass accuracy of less than 1 ppm.</li><li>Complete sgRNA sequence coverage (100%) was achieved by employing RapiZyme MC1, a novel endonuclease introduced by Waters, which provided unique cleavage specificity and predictable missed cleavages to generate longer, unique, overlapping digestion products.</li><li>Oligonucleotide isomeric sequences resulting from enzymatic digestion were effectively differentiated using the waters_connect CONFIRM Sequence App, which matched elevated energy fragment ions patterns to their respective sequences, allowing unambiguous isomer identification.</li></ul>

News from LabRulezLCMS Library - Week 42, 2025

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PFAS in Food & Food Contact Material