News from LabRulezLCMS Library - Week 31, 2025

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 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 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 10th November 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!</strong></p><h3><strong>1. Agilent Technologies: Feed Injection at Elevated Pressure Using the Agilent 1290 Infinity III Hybrid Multisampler</strong></h3><p><strong>Optimization of a multipesticide method for the highest sensitivity through feed injection</strong></p><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/te_feed_injection_multipesticide_1290_infinityiii_hybrid_5994_8478en_agilent_f070801ff2.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Modern analytical methods for the determination of pesticide residues are based on multipesticide methods capable of determining hundreds of pesticides in a sample in one analytical LC/MS/MS run. For chromatographic separation of these pesticides, modern separation columns are used. Their dimensions are typically 2.1 × 150 mm, filled with 1.9&nbsp;µm particle size material. This requires that the LC can work at high pressures up to 1300 bar. The Agilent 1290 Infinity III Hybrid Multisampler can operate within this pressure range. In addition, the Hybrid Multisampler can work in two different modes, the classic flow through mode and the Agilent Feed Injection mode. Feed Injection mode helps to prevent peak broadening and splitting effects as they can occur with the classic flow through mode for injection of samples in solvents with high elution strength and higher injection volumes.</p><h4><strong>Experimental</strong></h4><h5><strong>Instrumentation</strong></h5><ul><li>Agilent 1290 Infinity III High-Speed Pump (G7120A)&nbsp;</li><li>Agilent 1290 Infinity III Hybrid Multisampler (G7137B)&nbsp;</li><li>Agilent 1290 Infinity III MCT (G7116B)&nbsp;</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=Ultivo+Triple+Quadrupole+LC%2FMS+"><strong>Agilent Ultivo Triple Quadrupole LC/MS </strong></a>with Agilent Jet Stream source&nbsp;</li></ul><h5><strong>Software&nbsp;</strong></h5><ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=MassHunter+Acquisition+Software"><strong>Agilent MassHunter Acquisition Software</strong></a> (v. 12.2)&nbsp;</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=MassHunter+Qualitative+Analysis"><strong>Agilent MassHunter Qualitative Analysis Software</strong></a> (v. 12.0)&nbsp;</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=MassHunter+Quantitative+Analysis"><strong>Agilent MassHunter Quantitative Analysis Software</strong></a> (v.&nbsp;12.1)&nbsp;</li></ul><h5><strong>Column&nbsp;</strong></h5><ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=InfinityLab+Poroshell+120+Phenyl-Hexyl"><strong>Agilent InfinityLab Poroshell 120 Phenyl-Hexyl</strong></a>, 2.1 × 150 mm, 1.9 µm (part number 699675-912)</li></ul><h4><strong>Results and discussion&nbsp;</strong></h4><p>Calibration curves for a multipesticide method were acquired to demonstrate the possible improvement of peak shape and the lower limits of quantification (LOQ) using Agilent Feed Injection mode in comparison to classical flow through injection mode. This method comprised approximately 120&nbsp;pesticide compounds measured by dynamic MRM. The separation was performed on a 150 mm length column. With the chosen solvents and flow rate, a pressure range above 1000&nbsp;bar was necessary. The calibration curves were acquired for concentrations down to 0.02 ppb (20 ppt), and each calibration point was measured three-fold. The concentrations of the calibration solutions were prepared by dilution using the strong-eluting solvent acetonitrile and injected directly by both injection modes with different injection volumes. An example chromatogram showing the elution pattern for the MRM transitions is displayed in Figure 1.</p><h4><strong>Conclusion&nbsp;</strong></h4><p>This technical overview demonstrates the use of the Agilent Feed Injection mode available with the Agilent 1290 Infinity III Hybrid Multisampler in comparison to the classical flow through injection mode for the analysis of pesticides by a multipesticide method. We demonstrated that the use of Agilent Feed Injection mode improves the obtained peak shape for injection of samples dissolved in high elution strength solvent compared to classical flow through mode. With the improved peak shape obtained by Feed Injection mode, signal to noise ratios were improved, resulting in higher sensitivity. Due to retained peak shape, no manual interaction for peak integration was necessary when using feed injection.</p><h3><strong>2. Shimadzu: Automatic Optimization of Sample Solvent Composition for Achieving Sharp Peak Shapes</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_00938_en_c0f4de1265.pdf"><strong>Full PDF for download</strong></a></li></ul><h4><strong>User Benefits</strong></h4><ul><li>The automatic pretreatment function integrated into the autosampler enables efficient optimization of sample solvent composition, which can be automatically adjusted during analysis.</li><li>The desired composition of sample solvent can be easily specified via the LabSolutionsTM MD interface.</li></ul><p>In LC analysis, the composition of the sample solvent is critical for achieving proper peak shape. If the sample solvent is a stronger eluting solvent than the mobile phase, sample band condensation at the column inlet may be insufficient, leading to sample band broadening. For instance, increasing the organic solvent ratio in the sample solvent to dissolve low-polar compounds may deteriorate the peak shapes of early-eluting compounds in reversed-phase chromatography. Therefore, determining the optimal ratio of organic solvent in the sample solvent is essential. However, manually preparing multiple solvent compositions is extremely time-consuming. The automatic pretreatment function of the autosamplers (<a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=Nexera+X3"><strong>Nexera series</strong></a>), in combination with the method development support <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=LabSolutions+MD"><strong>software LabSolutions MD</strong></a>, enables consecutive analyses using different sample solvent compositions without manual intervention. This allows evaluation of the effect of sample solvent composition on peak shape while significantly reducing the labor involved in optimization. In this study, we demonstrate the automated determination of the optimal sample solvent composition using metoclopramide, a small-molecule drug, as a model compound.</p><h4><strong>Automatic Pretreatment Function</strong></h4><p>The automatic pretreatment function integrated into the autosampler enables drawing and ejecting specified volumes of reagents and solvents from any vial, as well as mixing them within the needle. As an example, Fig. 1 illustrates the procedure for automatically adjusting the water-to-methanol ratio in the sample solvent using this function. The autosamplers (Nexera series) supports up to three rinsing solvents via the multi-rinse function, and in this procedure, methanol and water were supplied from these rinsing lines. By modifying the "methanol eject volume" (2) and “water eject volume” (3) in Fig. 1, sample solvents with desired methanol-to-water ratios can be automatically prepared. In addition, adjusting the “sample drawing volume” (1) allows for not only changing the sample solvent composition but also diluting the sample solution to a specified concentration.</p><h4><strong>Analytical Conditions and Target Compounds</strong></h4><p>A 1000 mg/L solution of metoclopramide, a small-molecule drug, was prepared in 100% methanol as a model sample. The water content in the sample solvent was then varied from 0% to 90% in 10% increments (A 10-fold dilution was applied during automatic sample solvent preparation), and its effect on peak shape was evaluated under the analytical conditions shown in Table 1. By simply specifying the desired water content on the LabSolutions MD interface (red frame in Fig. 2), the autosampler’s automatic pretreatment function adjusts the sample solvent composition during consecutive analyses. This eliminates the need for manual preparation of multiple solvent mixtures, significantly reducing the workload involved in optimizing sample solvent composition for ideal peak shapes.</p><h4><strong>Conclusion</strong></h4><p>Optimizing the composition of the sample solvent is essential for achieving appropriate peak shapes. However, manual preparation of multiple solvent compositions is time-consuming. The automatic pretreatment function of the autosampler, in combination with LabSolutions MD, enables consecutive analyses with automatic variation of the sample solvent composition. This significantly reduces the effort required for optimization. Since the optimal sample solvent composition depends on both the physicochemical properties of the analyte and the mobile phase conditions, optimization is necessary for each sample. These tasks, however, can be greatly reduced by utilizing the automatic pretreatment function.</p><h3><strong>3. Thermo Fisher Scientific: Improving selectivity and sensitivity of lipid mediator analyses by coupling nano-flow chromatography with the Stellar mass spectrometer</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_improving_selectivity_sensitivity_lipid_mediator_analyses_po_en_c38bc37825.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Lipid mediators play crucial roles in regulating inflammation and maintaining tissue homeostasis. However, their quantification in biological samples is difficult due to low concentrations (ppt–ppb range), chemical instability, and structural similarity. Conventional LC-MS/MS methods using high-flow UHPLC and triple quadrupole mass spectrometers require extensive optimization of SRM transitions for each analyte. This study introduces a <strong>nano-flow LC-MS workflow</strong> combining the <strong>Thermo Scientific Vanquish Neo UHPLC system</strong> with the <strong>Thermo Scientific Stellar mass spectrometer</strong>, aiming to improve analytical sensitivity and selectivity while reducing solvent and sample consumption.</p><h4><strong>Experimental</strong></h4><p>Lipid mediator standards (from Cayman Chemical) were analyzed using two chromatographic configurations: a <strong>nano-flow method</strong> (500 nL/min) with the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/398"><strong>Vanquish Neo UHPLC</strong></a> and a <strong>high-flow method</strong> (300 µL/min) using the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/744"><strong>Vanquish Horizon UHPLC</strong></a>. Both setups were coupled either to the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1149"><strong>Stellar MS</strong></a> or the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/404"><strong>TSQ Altis Plus triple quadrupole MS</strong></a>. The Stellar instrument features a <strong>dual-pressure linear ion trap</strong> and <strong>ion routing multipole</strong>, enabling both <strong>HCD (higher-energy collisional dissociation)</strong> and <strong>CID (collision-induced dissociation)</strong> fragmentation. Parallel reaction monitoring (PRM) was employed instead of SRM, allowing simultaneous detection of multiple transitions. Data were processed using <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/77"><strong>Thermo Scientific TraceFinder</strong></a> software with internal standards and calibration curves ranging from 0.0001 ppb to 100 ppb.</p><h4><strong>Results</strong></h4><p>The nano-flow Stellar method achieved superior sensitivity and selectivity compared with the high-flow methods on both Stellar and TSQ Altis Plus systems. Limits of quantitation (LOQ) reached the <strong>low-ppt range</strong>, while maintaining fast analysis times (&lt; 30 min) and requiring less than 20 µL of solvent per sample. Chromatograms of key eicosanoids such as <strong>15(S)-HETE</strong>, <strong>12(S)-HETE</strong>, and <strong>5(S)-HETE</strong> demonstrated higher signal intensity and improved peak shapes under nano-flow conditions. Calibration curve reproducibility and scan density (&gt; 10 scans per peak) confirmed reliable quantitative performance.</p><h4><strong>Conclusion</strong></h4><p>Coupling <strong>nano-flow chromatography</strong> with the <strong>Stellar mass spectrometer</strong> provides a powerful workflow for targeted lipid mediator analysis, significantly enhancing both selectivity and sensitivity compared with conventional high-flow LC-MS/MS. The system’s <strong>PRM capability</strong>, <strong>dual-fragmentation flexibility</strong>, and <strong>low-volume solvent use</strong> make it a robust and cost-effective solution for biological and clinical lipidomics, capable of quantifying trace-level lipid mediators efficiently and reproducibly.</p><h3><strong>4. Waters Corporation: Investigating Drug Metabolism of Methapyrilene Within a Rat Model Using a Data Dependent Acquisition Workflow with the Xevo MRT Mass Spectrometer</strong></h3><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/720009001_en_c11ab3b6b7.pdf"><strong>Full PDF for download</strong></a></li></ul><h5><strong>Benefits&nbsp;</strong></h5><ul><li>Demonstration of the Waters™ Xevo™ MRT Mass Spectrometer (MS) workflow for the analysis of urine extracts from a drug dosed study, involving methapyrilene.</li><li>Data dependent acquisition (DDA) has been used for data collection, providing highly informative fragmentation data to confidently identify drug metabolites of interest.</li><li>Combining a DDA approach with high-throughput chromatography is shown to consistently yield high quality spectra, providing highly confident compound identifications.</li><li>A seamless workflow allowing drug metabolites to be confidently identified is demonstrated with the UNIFI™ Application within the waters_connect™ Software Platform</li></ul><p>The introduction of the Xevo MRT Mass Spectrometer allows for comprehensive drug metabolite identification (Met ID) at high acquisition speed. Using urine samples collected over a time course of six days, the excellent mass accuracy, high mass resolution, and sensitivity are highlighted, which can be achieved for drug metabolism studies. DDA provides highly specific MS/MS information, which assists with structural elucidation. The speed at which DDA data can be acquired with the Xevo MRT Mass Spectrometer can be as fast as 50 and 100 Hz, MS, and MS/MS respectively. Combined with fast chromatographic methods, this fast data acquisition rate allows for highly informative, confident data in a high-throughput fashion</p><h4><strong>Experimental</strong></h4><p>LC-MS Data Acquisition LC-MS data were collected using an <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/191"><strong>ACQUITY™ Premier UPLC™ System</strong></a> coupled to a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1148"><strong>Xevo MRT Mass Spectrometer</strong></a>. The reversed-phase (RP) chromatographic separation consisted of a 5.0 minute method (injection to injection) using a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1966"><strong>CORTECS™ C18</strong></a>, 1.6 µm, 2.1 x 50 mm analytical Column (p/n: 186007114). The LC solvents were comprised of mobile phase A (0.1% formic acid containing 1 mMol ammonium formate) and mobile phase B (95:5 ACN:Water (v/v), 0.1% formic acid, 1 mMol ammonium formate) being ramped from 50% to 99% over 5 minutes using a flow rate of 0.6 mL/minute. The gradient started from 0.1% to 15% (mobile phase B) over 1.5 minutes, 50% at 3 minutes and 99% at 4 minutes. The solvent composition was held at 99% (mobile phase B) for 0.5 minutes and re-equilibration at initial conditions by 5 minutes, ready for the subsequent injection.&nbsp;</p><p>The Xevo MRT Mass Spectrometer was configured with the source settings as outlined in Table 1. MS data were collected using DDA mode of operation, which consisted of scan rates of 20 Hz (MS) and 50 Hz (MS/MS). The number of components selected for MS/MS were eight with a dynamic exclude applied of 10 mDa with a 10 second retention time tolerance.</p><h4><strong>Data Processing&nbsp;</strong></h4><p>The acquired data were processed using the UNIFI Application within the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1544"><strong>waters_connect Software Platform</strong></a>. Data were peak picked and screened against a compound database containing MP metabolites and their associated fragments. A mass tolerance of 1 ppm for both MS and MS/MS were applied.</p><h4><strong>Conclusion&nbsp;</strong></h4><p>LC-MS analysis of urine collected over a time course following administration of the drug methapyrilene, has been demonstrated using a DDA approach with the Xevo MRT Mass Spectrometer. MP undergoes complex biotransformation to provide a variety of metabolites. Utilizing a DDA workflow provides detailed, clean fragment ion spectra for confident structural elucidation. The additional benefit of sub ppm mass accuracy provides definitive identification(s). Integrating fast chromatography allows for high-throughput analyses while maintaining high resolution and mass accuracy.</p>

News from LabRulezLCMS Library - Week 46, 2025

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News from LabRulezLCMS Library - Week 02, 2026

<p><strong>Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 12th January 2026? Check out new documents from the field of liquid phase, especially HPLC and LC/MS techniques!</strong></p><blockquote><p><span style="font-size:20px;">👉 </span><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library"><span style="font-size:20px;"><strong>SEARCH THE LARGEST REPOSITORY OF DOCUMENTS ABOUT LCMS AND RELATED TECHNIQUES</strong></span></a></p></blockquote><blockquote><p><strong>👉 Need info about different analytical techniques?</strong> Peek into <a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/library"><strong>LabRulezGCMS</strong></a> or <a target="_blank" rel="noopener noreferrer" href="https://icpms.labrulez.com/library"><strong>LabRulezICPMS</strong></a> libraries.</p></blockquote><p>This week we bring you application notes by <strong>Agilent Technologies, Shimadzu, Waters Corporation </strong>and poster by<strong> Thermo Fisher Scientific / HPLC!</strong></p><h3><strong>1. Agilent Technologies: B Vitamin Analysis in Dietary Supplements on the Agilent 1290 Infinity III LC System</strong></h3><p><strong>Using ion-pairing reagent and supported by time‑saving automation of routine tasks using the Agilent InfinityLab Assist</strong></p><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/an_b_vitamin_dietary_supplements_1290_infinityiii_lc_5994_8826en_agilent_56b5eb2fb1.pdf"><strong>Full PDF for download</strong></a></li></ul><p>There is a huge variety of (multi)vitamin supplements on the market, with different vitamin combinations and different concentrations available. To ensure that the claimed vitamin contents are met, analysis of those supplements is&nbsp;necessary.&nbsp;</p><p>To ensure constant and good analytical performance, preparation of the analytical system is fundamental. There are different approaches to preparing the system for analysis. Purging the solvent lines from bottle to pump removes possible air bubbles, especially after longer breaks between instrument usage or changing eluent to the ion-pairing and buffer-containing eluent after the weekend. Equilibration can either be done by a calculated time (10–20 column volumes)1 , set as a fixed experimentally determined value, or manually monitored. Using a fixed time for equilibration can lead to unnecessary high solvent consumption, as equilibration may already be established and no monitoring was previously done. In contrast, manual monitoring of equilibration can be time-consuming and error-prone. For example, if there are no written rules regarding key points for equilibration of the column, its determination can be subjective (constant&nbsp;pressure and baseline of e.g. UV absorption).&nbsp;</p><p>As vitamins are grouped according to their water solubility and not their chemical structure, separation behavior can greatly differ as they have different charges at physiological pH. In such cases, ion-pairing reagents such as hexane sulfonic acid can enable the separation of those vitamins while using a reversed phase column. As ion-pairing LC depends on good equilibration to ensure the presence of ion‑pairing reagent within the system, the process of preparing the instrument is an inevitable step. This is essential for good and reproducible analytical results.&nbsp;</p><p>With the Agilent InfinityLab Assist, making your instrument ready is no longer a time-consuming and unpleasant procedure.<sup>1</sup> This application note demonstrates the separation of multiple B vitamins in multivitamin tablets and effervescent tablets using an <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?search=1290+Infinity+III"><strong>Agilent 1290 Infinity III LC System</strong></a>. Furthermore, it is shown how the InfinityLab Assist can help users ensure good chromatographic results, save time, and reduce errors by replacing manual tasks with InfinityLab Assist tasks.&nbsp;</p><h4><strong>Experimental&nbsp;</strong></h4><h5><strong>Equipment&nbsp;</strong></h5><ul><li>Agilent 1290 Infinity III High-Speed Pump (G7120A)&nbsp;</li><li>Agilent 1290 Infinity III Multisampler (G7167B) with Agilent InfinityLab Sample Thermostat (G4761A)&nbsp;</li><li>Agilent 1290 Infinity III Multicolumn Thermostat (G7116B) with Agilent InfinityLab Quick Connect Heat Exchanger (G7116-60015)&nbsp;</li><li>Agilent InfinityLab Level Sensing (G7175A)&nbsp;</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=InfinityLab+Assist"><strong>Agilent InfinityLab Assist</strong></a> (G7180A) with Agilent InfinityLab Assist Control Software, version 2.0&nbsp;</li><li>Agilent 1290 Infinity III Diode Array Detector (G7117B) with Agilent InfinityLab Max-Light Cartridge Cell (10 mm, 1.0 µL, G4212-60008)&nbsp;</li></ul><h5><strong>Software&nbsp;</strong></h5><ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=OpenLab+CDS"><strong>Agilent OpenLab CDS,</strong></a> version 2.8, update 7 (or later versions)&nbsp;</li></ul><h5><strong>Column&nbsp;</strong></h5><ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=10&amp;search=InfinityLab+Poroshell+120+EC-C18"><strong>Agilent InfinityLab Poroshell 120 EC-C18</strong></a>, 2.1&nbsp;mm × 100 mm, 2.7 µm column (p/n 695775-902).</li></ul><h4><strong>Results and discussion&nbsp;</strong></h4><h5><strong>Separation of vitamin B1, B2, B5, B9, and B12&nbsp;</strong></h5><p>To analyze vitamin B1, B2, B5, B6, B7, B9, and B12 in vitamin supplements, different columns, mobile phases, and gradients were evaluated. The selected chromatographic conditions (Table 1) showed the best separation from peak to peak and additional matrix peaks. Hexane sulfonate was needed as an ion-pairing reagent to ensure good peak shapes, as all vitamins investigated, except vitamin B12, are expected to be charged based on their pKa values.3 The method detects the B vitamins investigated in vitamin tablets and effervescent tablets as demonstrated in Figure 1, showing chromatograms of aqueous standards and real samples. As some vitamins are present at higher doses, such as the 12 mg tablet (vitamin&nbsp;B5) compared to the 2.5 µg tablet (B12) in supplement 3 (Figure 1), parallel analysis is rather difficult, and not all vitamins present were detectable in each&nbsp;supplement.</p><h4><strong>Conclusion&nbsp;</strong></h4><p>Using the Agilent 1290 Infinity III LC System setup detection of vitamins B1, B2, B5, B6, B7, B9, and B12 in effervescent tablets and multivitamin supplement matrix, good separation was achieved. Using the Agilent InfinityLab Assist "Make&nbsp;Ready" task, the system was prepared for B vitamin analysis using only one automated task and no manual monitoring. Running this task automatically by scheduling saved more than 20 minutes of working time. Additionally, the time required for medium equilibration monitoring sensitivity was reduced by approximately six minutes, compared to manual equilibration. Furthermore, using the schedulable InfinityLab Assist "Standby" task, the risk of errors regarding instrument standby can be drastically reduced. User-defined settings enable adjustment for different purposes, such as switching of all modules except the sampler thermostat.<sup>2</sup> The ability to remove the used buffer and ion-pairing reagent from the flow path for longer storage by scheduling a standby task also greatly helps to prevent salt precipitation in the system without additional effort. Also, the InfinityLab Assist can clearly help to accelerate checking the instrument status while preparing the instrument, as well as during running&nbsp;samples.<sup>2</sup></p><h3><strong>2. Shimadzu: Analysis of Ammonia as an Impurity in Fuel Cell Grade Hydrogen According to ISO 14687 Grade D Using a Gas Generator</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_01070_en_43a535bf87.pdf"><strong>Full PDF for download</strong></a></li></ul><p>Hydrogen is expected to be a new power and heat source to reduce greenhouse gas emissions. Hydrogen is an easily available substance that can be produced from the electrolysis of water in our daily lives. However, when used in fuel cells, it is necessary to control impurities in hydrogen gas because certain impurities introduced during the manufacturing process deteriorate the electrode and electrolyte membrane, reducing the performance of the fuel cell or shortening its life. When ammonia is used as a carrier of hydrogen gas due to its distribution as a chemical, there is a possibility that ammonia may be introduced into hydrogen. This article presents an example of analysis of ammonia in hydrogen using an ion chromatograph (IC) based on the standard values described in ISO 14687 Grade D.<sup>1)</sup></p><h4><strong>Analysis of Ammonium Ion by IC</strong></h4><p>A <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=1&amp;search=HIC-NS"><strong>non-suppressor IC (HIC-NS)</strong></a> was used to analyze NH<sub>4</sub>. Since the IC can selectively measure ions dissolved in the sample, it is less affected by contaminants than other methods and can measure NH<sub>4</sub> with high accuracy. In addition, by using the same device together with a suppressor IC, it is possible to measure inorganic halogens and formic acid, which are other impurities listed in ISO 14687. Table 1 shows the analytical conditions for NH<sub>4</sub>, Fig. 2 shows the chromatogram of a standard sample at a concentration near the specified value (25 μg/L), and Fig. 3 shows the calibration curve in the range from 2.5 μg/L to 200 μg/L. As shown in Fig. 3, the linearity was greater than 0.9999. Table 2 shows the reproducibility of the sample at 10 μg/L, which is around 1/2 of the specified concentration, repeated 7 times.</p><h4><strong>Conclusion</strong></h4><p>Ammonia, an impurity in hydrogen, was analyzed using a nonsuppressor IC, and it was confirmed that the method met the standard value specified by ISO 14687 Grade D. In addition, when ammonia standard gas near the standard value was mixed into hydrogen using a gas generator, a good recovery rate was obtained, which confirmed the validity of the method including the collection flow of pretreatment.</p><h3><strong>3. Thermo Fisher Scientific / HPLC: High-sensitive multi-attribute analysis of ADCs under native conditions by using an online multiple heart-cut 2D-LC-HRAM mass spectrometry system</strong></h3><ul><li>Poster</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_high_sensitive_multi_attribute_analysis_adcs_native_conditions_po_en_28e7e0b002.pdf"><strong>Full PDF for download</strong></a></li></ul><p>ADCs are a rapidly expanding class of biotherapeutics for cancer treatment. Critical quality attributes such as charge heterogeneity, DAR, and drug distribution are essential for ensuring the safety, stability, and efficacy of ADCs. Strong cation exchange (SCX) and hydrophobic interaction chromatography (HIC) are conventional methods employed for charge variant analysis and DAR determination of ADCs1. However, these methods frequently utilize MS-incompatible salts, limiting their use for MS characterization. Online 2D-LC addresses this by enabling desalting in the second dimension(2D) and offering multi-attribute analysis in one method. Here, multiple heart-cut 2D-LC-HRAM MS methods were developed to achieve high sensitivity for the multi-attribute analysis of polatuzumab vedotin under native conditions.</p><h4><strong>Methods</strong></h4><p>A <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/742"><strong>Thermo Scientific Vanquish 2D UHPLC system</strong></a>, coupled with a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/402"><strong>Thermo Scientific Orbitrap Exploris 240 mass spectrometer</strong></a> with Biopharma option, were utilized to configure the multiple heart-cut 2D-LC-HRAM MS system. HIC-SEC and SCXSEC methods were developed and optimized for the charge variants, drug-to-antibody ratio (DAR) and drug load distribution (DLD) characterization of ADCs under native conditions.</p><h4><strong>Conclusions&nbsp;</strong></h4><p>In this study, an online multiple heart-cut 2D-LC-HRAM MS system was configured and optimized for the multi-attribute analysis of polatuzumab vedotin. This approach provides detailed and comprehensive insights into the charge heterogeneity analysis, DAR, and DLD analysis of polatuzumab vedotin under native conditions.&nbsp;</p><ul><li>The online multiple heart-cut 2D-LC system enables the direct transfer of SCX and HIC fractions to mass spectrometry analysis by utilizing SEC as a desalting step, thereby overcoming the MS-incompatibility buffer in the LC separation.&nbsp;</li><li>The backflush transfer of fractions was employed to enhance peak shape and sensitivity in the 2D SEC separation.&nbsp;</li><li>The 2D SEC-MS analysis enables the characterization of ADCs under native conditions.&nbsp;</li><li>The high sensitivity of the Orbitrap Exploris 240 MS allows for the identification of fractions with low abundance in the 1D chromatography</li></ul><h3><strong>4. Waters Corporation: Automated Multiple Reaction Monitoring (MRM) Method Development for Peptide Drugs Using waters_connect for Quantitation Software</strong></h3><ul><li>Application note</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/720009123_en_1f51e50be8.pdf"><strong>Full PDF for download</strong></a></li></ul><h5><strong>Benefits&nbsp;</strong></h5><ul><li>Rapid evaluation and optimization of MRM conditions freeing up bioanalysts to perform other valuable tasks</li><li>Interactive graphical review of results facilitating informed decision making.&nbsp;</li><li>Elimination of transcription errors via direct transfer of MRM results to the LC-MS method editor.</li></ul><p>LC-MS/MS is the technology of choice for the quantification of drugs, metabolites, and biomarkers in biofluids, typically employing tandem quadrupole mass spectrometry (MS) with detection via MRM. MRM method development for classic small molecule drugs involves the selection of the best precursor and product ion followed by optimization of cone voltages and collision energies. This is achieved by infusion or LC injection of the analyte(s) of interest. For small molecule drug compounds, this process is usually straightforward and lends itself to automation as they largely generate singly charged ions. However, in electrospray mass spectrometry, biomolecules such as peptides, oligonucleotides, and proteins normally form several multiply charged precursor ions, e.g., M+3H3+, M+4H4+. This significantly complicates MRM method development, as each charge state can generate multiple fragment ions, all of which need to be compared both within and across each charge state to choose the best transition or transitions for analyte measurement.&nbsp;</p><p>The MRM Optimization tool with the waters_connect for Quantification Software has been specifically designed to address the challenge of MRM method development for multiply charged biomolecules. The Optimization tool facilitates the automatic acquisition of data from multiple precursor and product ion and collision energy combinations across several charge states, with the results reviewed via an interactive graphical interface. Here, the use of this optimization is illustrated, using the GLP-1 peptide, semaglutide, as an example.</p><h4><strong>Experimental</strong></h4><h5><strong>Mass Spectrometry</strong></h5><p>Mass spectrometry was performed on a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/2100"><strong>Waters Xevo™ TQ Absolute XR Mass Spectrometer</strong></a>, operated in positive ion electrospray ionization mode (ESI). Nitrogen was used as the nebulizer gas and argon as the collision gas. The cone voltage, collision energy, and MS/MS transitions were adjusted and evaluated using the waters_connect MRM Optimization tool.</p><h5><strong>Informatics&nbsp;</strong></h5><p>The mass spectrometer was controlled and resulting data evaluated using <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1544"><strong>waters_connect for Quantitation</strong></a> Application Manager version 1.9.</p><h4><strong>Conclusion&nbsp;</strong></h4><p>The number of candidate biomolecule drugs, peptides, oligonucleotides, and proteins in pharmaceutical discovery and development is steadily increasing. These biomolecules require the development of quantitative LC-MS/MS assays to support discovery, preclinical, and human DMPK studies. Unlike small molecules, these biomolecules form multiply charged ions in the MS source, making MRM method development more challenging, as multiple charge state–product ion combinations must be evaluated and compared. The Optimization tool in waters_connect for Quantitation Software provides an automated, fast platform for the optimization and evaluation of the precursor ion → product ion pairs formed at each charge state, simplifying the process of LCMS/MS method development for biomolecules.</p>

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