🎤Dr. Christopher Reid (Professor of Chemistry and Biochemistry at Bryant University in Rhode Island)In this episode of Concentrating on Chromatography, host Dave Oliva sits down with Chris Reid to explore how chemical biology and LC-MS are being used to tackle one of the most urgent global challenges: antimicrobial resistance.As antibiotic resistance continues to rise—and new drug development struggles to keep pace—researchers are looking beyond traditional approaches. Instead of targeting how bacteria build their cell walls, Chris’s work focuses on how they break them down, opening the door to entirely new antimicrobial strategies.<h4>What You’ll Learn in This Episode:</h4><ul><li>Why antimicrobial resistance is accelerating globally</li><li>What peptidoglycan is and why it’s essential to bacterial survival</li><li>How targeting cell wall degradation differs from traditional antibiotics</li><li>The strategy behind designing small molecule probes and inhibitors</li><li>How LC-MS is used to analyze peptidoglycan fragments and enzyme activity</li><li>Key analytical challenges: complex mixtures, low-abundance species, and structural similarity</li><li>The importance of sample preparation and reproducibility in LC-MS workflows</li><li>How this research could lead to new antibiotic targets</li></ul><h3>Video Transcription</h3><h4>The Growing Challenge of Antibiotic Resistance</h4>According to Reid, antibiotic resistance is increasing across a broad spectrum of bacterial pathogens that were once readily treated with existing therapies. In some cases, clinicians must now rely on only a handful of last-line antibiotics, while resistance is also rising in diseases such as tuberculosis.One of the major obstacles is the limited commercial incentive to develop entirely new antibiotics. Because bacterial resistance can emerge relatively quickly after a drug reaches the market, pharmaceutical companies often favor modifying existing antibiotics or combining them with resistance-modifying agents rather than investing in completely new drug classes. As a result, genuinely novel antibacterial targets remain urgently needed.<h4>Why the Bacterial Cell Wall Remains an Attractive Target</h4>The bacterial cell wall consists primarily of peptidoglycan, a highly cross-linked polymer that provides structural integrity, maintains cell shape, and enables bacterial growth and division.Many widely used antibiotics—including the β-lactam antibiotics such as penicillins—target enzymes responsible for synthesizing or cross-linking peptidoglycan. These pathways have proven highly successful therapeutic targets because peptidoglycan is unique to bacteria and absent from human cells, minimizing unwanted effects on the host.However, most existing antibiotics target only specific stages of peptidoglycan biosynthesis. Reid's research explores another part of this complex pathway: enzymes responsible for degrading the carbohydrate backbone of peptidoglycan during normal bacterial growth and remodeling. These enzymes may represent an underexplored class of antimicrobial targets capable of overcoming existing resistance mechanisms.<h4>Targeting the Glycan Backbone</h4>A key advantage of focusing on the carbohydrate backbone is its remarkable conservation across both Gram-positive and Gram-negative bacteria.Whereas the peptide components of peptidoglycan vary considerably between bacterial species—and frequently acquire mutations associated with antibiotic resistance—the glycan backbone remains highly conserved and tolerates relatively few structural modifications.This biological constraint makes enzymes acting on the glycan backbone particularly attractive targets. Reid's group developed a series of small-molecule probes and inhibitors designed to recognize enzymes known as N-acetylglucosaminidases, which cleave glycosidic bonds within peptidoglycan and generate soluble degradation products.Rather than beginning with purified enzymes, the researchers adopted a top-down phenotypic screening strategy. Candidate compounds were first evaluated for their ability to inhibit bacterial growth before progressively narrowing the list of potential molecular targets through genetic studies, microscopy, enzyme assays, and biochemical characterization. This iterative workflow ultimately identified compounds capable of inhibiting the desired enzyme class while simultaneously providing valuable chemical probes for studying bacterial cell wall biology.<h4>LC-MS as a Central Analytical Tool</h4>Liquid chromatography coupled with mass spectrometry plays a central role throughout the project by enabling detailed characterization of structural changes occurring within peptidoglycan after bacterial cells are treated with candidate inhibitors.Because intact peptidoglycan is an insoluble polymer, sample preparation begins with isolation of the bacterial cell wall through detergent extraction and differential centrifugation. Additional purification steps remove proteins, nucleic acids, and teichoic acids before enzymatic digestion converts the insoluble material into soluble fragments suitable for chromatographic analysis.The resulting digest is analyzed by LC-MS to examine changes in the distribution of peptidoglycan fragments. Chromatographic retention times provide an immediate indication of changes in cross-linking, with larger oligomers such as trimers and tetramers generally eluting later than monomeric fragments. Mass spectrometry then confirms the identity of these fragments by measuring their intact molecular masses, while tandem MS experiments can provide further structural information when required.<h4>Sample Preparation Determines Data Quality</h4>Reid emphasized that sample preparation represents one of the most critical—and challenging—stages of the entire workflow.Because peptidoglycan analysis involves multiple purification, digestion, and reduction steps, even minor procedural differences between samples can introduce analytical artifacts that complicate biological interpretation.Maintaining strict consistency throughout sample preparation is therefore essential. Appropriate controls must accompany every experiment, ensuring that observed chromatographic and mass spectrometric differences truly reflect biological responses rather than experimental variability. The complexity of bacterial cell wall composition further increases the analytical challenge, as subtle differences in amino acid substitutions and low-abundance cross-linked species can complicate LC-MS data interpretation.<h4>Chemical Biology for Future Antibiotic Discovery</h4>Although the identified compounds may eventually serve as starting points for antibiotic development, Reid views them primarily as chemical biology tools.By selectively inhibiting individual enzymes, these probes allow researchers to investigate previously unexplored aspects of bacterial cell wall remodeling that cannot easily be studied using classical genetic approaches alone.The research team is now expanding this strategy by combining their probes with proximity-labeling proteomics and mass spectrometry, aiming to map protein interaction networks associated with bacterial cell wall degradation. Such studies may reveal entirely new antibacterial targets and provide deeper insight into the biological processes governing bacterial growth and division.Ultimately, the work illustrates how integrating synthetic chemistry, microbiology, molecular biology, and advanced LC-MS can accelerate the search for innovative therapeutic strategies against antimicrobial resistance. As existing antibiotics become progressively less effective, multidisciplinary approaches such as these may play an increasingly important role in identifying the next generation of antibacterial targets.<blockquote>This text has been automatically transcribed from a video presentation using AI technology. It may contain inaccuracies and is not guaranteed to be 100% correct.</blockquote><blockquote><h4>Concentrating on Chromatography Podcast</h4>Dive into the frontiers of chromatography, mass spectrometry, and sample preparation with host David Oliva. Each episode features candid conversations with leading researchers, industry innovators, and passionate scientists who are shaping the future of analytical chemistry. From decoding PFAS detection challenges to exploring the latest in AI-assisted liquid chromatography, this show uncovers practical workflows, sustainability breakthroughs, and the real-world impact of separation science. Whether you’re a chromatographer, lab professional, or researcher you'll discover inspiring content!You can find Concentrating on Chromatography Podcast in podcast apps:<ul><li><a target="_blank" rel="noopener noreferrer" href="https://open.spotify.com/show/1wZg67rCjoGju4yhqxI3tL">Spotify</a></li><li><a target="_blank" rel="noopener noreferrer" href="https://podcasts.apple.com/tz/podcast/concentrating-on-chromatography/id1788429185">Apple podcast</a></li><li><a target="_blank" rel="noopener noreferrer" href="https://podscan.fm/podcasts/concentrating-on-chromatography">podscan</a></li></ul>and on <a target="_blank" rel="noopener noreferrer" href="https://www.youtube.com/@ChromatographyTalk/videos">YouTube channel</a></blockquote>

 Fighting Antibiotic Resistance with LC-MS: Targeting Bacterial Cell Walls

This study compares a commercial ELISA assay with a validated UHPLC-MS/MS method for determining total ergot alkaloids in wheat and durum samples. In addition to method performance, the authors investigated cross-reactivity and matrix effects to explain differences between immunoassay and chromatographic results.While ELISA demonstrated good repeatability and is well suited for high-throughput screening, systematic biases were observed due to compound- and matrix-dependent cross-reactivity. The findings support ELISA as an effective screening tool, whereas UHPLC-MS/MS remains the preferred method for accurate quantification and regulatory confirmation of ergot alkaloids.<blockquote><h4>The original article</h4><h4><a target="_blank" rel="noopener noreferrer" href="https://pubs.acs.org/doi/10.1021/acs.jafc.6c05234">Comparative Evaluation of Total Ergot Alkaloids Determination by ELISA and UHPLC-MS/MS in Wheat: Implications of Cross-Reactivity and Matrix Effects</a></h4>Chamali Kodikara*, Dainna Drul, Nandika Bandara, and Sheryl A. Tittlemier*J. Agric. Food Chem. 2026, 74, 25, 20015–20027<a target="_blank" rel="noopener noreferrer" href="https://doi.org/10.1021/acs.jafc.6c05234">https://doi.org/10.1021/acs.jafc.6c05234</a><h6>licensed under <a target="_blank" rel="noopener noreferrer" href="https://creativecommons.org/licenses/by/4.0/">CC-BY 4.0</a></h6></blockquote>Selected sections from the article follow. Formats and hyperlinks were adapted from the original.<div>Ergot alkaloids (EAs) are a structurally diverse group of indole-derived mycotoxins produced primarily by phytopathogenic fungi of the genus Claviceps, most notably Claviceps purpurea. These fungi infect the flowering heads of cereal crops, replacing the developing grain with hardened fungal structures (sclerotia) that contain ergot alkaloids. Cereals such as rye, wheat, barley, oats, and triticale are particularly susceptible, with infection severity strongly influenced by environmental conditions during flowering, including rainfall and humidity. <a href="javascript:void(0);">(1−3)</a> Although improvements in agronomic practices and grain cleaning have reduced the prevalence of visible ergot sclerotia, ergot alkaloids can persist in cereals and cereal-based foods even in the absence of detectable sclerotia. This may occur because sclerotia can fragment during harvesting, handling, cleaning, or milling, producing small particles that are not visually detected but may still contribute ergot alkaloids to grain or processed products, continuing to pose food and feed safety concerns. <a href="javascript:void(0);">(4)</a> <a href="javascript:void(0);">(5)</a> Ergot alkaloids share a common tetracyclic ergoline backbone and occur as structurally related compounds, including ergometrine, ergotamine, ergosine, ergocristine, ergocryptine, and ergocornine, together with their corresponding C-8 epimers. These epimers occur as R- and S-configurations, commonly referred to as -ine and -inine forms, respectively. Epimerization at the C-8 position can occur during fungal metabolism and during sample extraction, storage, and analysis, complicating accurate quantification. <a href="javascript:void(0);">(6,7)</a> Although S-epimers were historically considered biologically inactive, recent evidence indicates that they may contribute to overall exposure and toxicity, supporting their inclusion in analytical and regulatory frameworks. <a href="javascript:void(0);">(3)</a>Dietary exposure to elevated levels of ergot alkaloids has been associated with a range of adverse health effects, collectively referred to as ergotism, including vasoconstriction, ischemia, neurological disturbances, and hallucinations. <a href="javascript:void(0);">(3)</a> These effects arise primarily from interactions with adrenergic, serotonergic, and dopaminergic receptors. <a href="javascript:void(0);">(8−10)</a> In response, regulatory limits for ergot alkaloids in food have been introduced or revised in several jurisdictions. In the European Union, maximum levels have been established for the sum of 12 ergot alkaloids in cereal milling products under Regulation (EU) 2021/1399, which was subsequently consolidated under Regulation (EU) 2023/915. This sum includes ergometrine, ergosine, ergotamine, ergocornine, ergocryptine, and ergocristine together with their corresponding -inine epimers, increasing the demand for reliable analytical methods to support routine monitoring and enforcement. <a href="javascript:void(0);">(1,11)</a>Liquid chromatography-tandem mass spectrometry (LC-MS/MS), particularly ultrahigh performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), is widely regarded as the reference technique for confirmatory determination of ergot alkaloids due to its sensitivity, selectivity, and ability to resolve epimeric forms. Recent methods routinely achieve low microgram-per-kilogram limits of quantification across a range of cereal matrices. <a href="javascript:void(0);">(12−17)</a> However, LC-MS/MS approaches are resource-intensive and typically confined to centralized laboratories, limiting their suitability for high-throughput screening or rapid decision-making in grain-handling and quality-control settings. Consequently, rapid screening methods, most notably enzyme-linked immunosorbent assays (ELISAs), are commonly employed as complementary tools to chromatographic analysis. <a href="javascript:void(0);">(18−21)</a> ELISAs offer reduced analysis time, simplified workflows, and lower per-sample costs, making them attractive for preliminary screening and sample triage. <a href="javascript:void(0);">(22−24)</a>In contrast to other regulated mycotoxins, however, the commercial availability of ELISAs for ergot alkaloids is limited. While multiple kits have been evaluated, performance has varied substantially across assays and matrices. <a href="javascript:void(0);">(1)</a> At present, the Randox Food Diagnostics Ergot Alkaloids ELISA is the only widely marketed microplate ELISA that quantifies the sum of 12 major ergot alkaloids, comprising ergometrine, ergosine, ergotamine, ergocornine, ergocryptine, and ergocristine together with their corresponding -inine epimers, consistent with the analyte scope used for EU maximum levels for ergot alkaloids in specified cereal products. <a href="javascript:void(0);">(25)</a>In addition, cereal matrix effects may influence antibody binding and signal generation, potentially resulting in biased quantification relative to LC-MS/MS reference methods. These limitations are particularly relevant given the compositional variability of ergot alkaloids across cereal species, geographic regions, and growing conditions. <a href="javascript:void(0);">(12)</a> Recent reviews emphasize the need for systematic, matrix-specific evaluation of immunoassay performance before such methods can be confidently integrated into regulatory or industrial monitoring programs. <a href="javascript:void(0);">(1,6,11)</a> However, independent evaluations of commercially available ergot alkaloid ELISAs, particularly those benchmarked against validated UHPLC-MS/MS methods using naturally infested wheat samples, remain limited.In this context, we hypothesized that measurements of total ergot alkaloids obtained using a commercially available ELISA are influenced by matrix effects and antibody cross-reactivity, resulting in systematic differences compared to UHPLC-MS/MS, which provides compound-specific quantification. To test this, the objective of the present study was to conduct a comprehensive performance evaluation of a commercially available ELISA for the detection of ergot alkaloids in wheat, using UHPLC-MS/MS as a reference method. Specific aims included assessment of repeatability and intermediate precision, extraction efficiency and extract stability, the influence of test portion mass, method accuracy relative to UHPLC-MS/MS, and characterization of antibody cross-reactivity toward individual ergot alkaloids in both solvent and wheat matrix. The results provide data-driven guidance on the appropriate use of ELISA-based screening within tiered analytical strategies for regulatory monitoring and food safety decision-making.<h4>2. Materials and Methods</h4><h5>2.3. UHPLC-MS/MS Reference Method</h5>UHPLC-MS/MS analysis of ergot alkaloids was performed using the validated Canadian Grain Commission (CGC) method described by Tittlemier et al. (2015). Briefly, ground grain (10 g) was extracted with 50 mL of acetonitrile/aqueous ammonium carbonate (84:16, v/v; 3.03 mM ammonium carbonate) to provide alkaline extraction conditions, then shaken for 30 min and centrifuged at 2450g for 10 min at room temperature. An aliquot of extract (464 μL) was combined with internal standard solution (dihydroergotamine; 20 μL of 4 ng/μL) and aqueous ammonium carbonate (1536 μL), vortex-mixed, and filtered through a 0.45 μm polytetrafluoroethylene syringe filter before injection into the UHPLC-MS/MS system. As in the validated reference method, dihydroergotamine was added after extraction to correct for postextraction analytical variability; extraction recovery was evaluated during method validation. Chromatographic separation was achieved using a reversed-phase BEH C18 column (2.1 × 100 mm, 1.7 μm; Waters, Milford, MA, USA) with the autosampler maintained at 15 °C and the column at 30 °C. Mobile phases consisted of (A) 3.03 mM aqueous ammonium carbonate and (B) acetonitrile. The gradient program was as follows: initial, 75% A/25% B; 1.00 min, 60% A/40% B; 5.00 min, 40% A/60% B; 8.00 min, 22% A/78% B; 10.50–11.00 min, 10% A/90% B; and 13.00–16.00 min, 75% A/25% B, with a total run time of 16 min. Detection was performed using positive electrospray ionization in multiple reaction monitoring mode on a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/554">Waters Xevo triple quadrupole mass spectrometer</a> (Waters, Milford, MA, USA). Under the validated chromatographic conditions, dihydroergotamine did not coelute with ergotamine, ergotaminine, or the other target ergot alkaloids. Identification and quantification were based on retention-time criteria and compound-specific MRM transitions, thereby minimizing potential isotopic or coelution interference. Precautions to minimize epimerization, including controlled handling of standards and extracts and reduced light exposure, were followed as described in the reference method. UHPLC-MS/MS quantification was performed using analyte-specific, matrix-matched calibration curves prepared according to the validated CGC reference method, with calibration levels of 0.046, 0.2, 1, 2, 10, and 20 μg/kg. Dihydroergotamine was used as the internal standard, and concentrations were calculated from internal-standard-normalized analyte responses with method dilution factors applied. Total ergot alkaloid concentration was calculated as the arithmetic sum of all quantified ergot alkaloids and epimers included in the validated method. <a href="javascript:void(0);">(26)</a><h4>3. Results and Discussion</h4><h5>3.4. Comparison of ELISA and UHPLC-MS/MS Results</h5>For method comparison, UHPLC-MS/MS was used as the chemical reference method, providing compound-specific quantification of individual ergot alkaloids and their epimers. A total ergot alkaloid concentration was calculated as the arithmetic sum of all quantified alkaloids included in the method. This summed value represents a chemically defined total based on compound-specific quantification of each analyte, rather than an immunoreactivity-weighted response governed by antibody affinity. As such, the UHPLC-MS/MS total serves as a chemically defined comparator against which the immunoreactivity-weighted ELISA response can be interpreted. Agreement between the commercial ELISA and the validated UHPLC-MS/MS reference method was evaluated using wheat and durum wheat cargo samples (n = 102) collected over multiple crop years (<a target="_blank" rel="noopener noreferrer" href="https://pubs.acs.org/doi/10.1021/acs.jafc.6c05234#fig5">Figure 5</a>). Linear regression analysis demonstrated a positive association between ELISA and UHPLC-MS/MS results across the full concentration range, indicating that the ELISA was responsive to increasing ergot alkaloid levels. However, the regression slope deviated substantially from unity, with ELISA results systematically lower than UHPLC-MS/MS values, indicating an approximate 2-fold underestimation.&nbsp;<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/J_Agric_Food_Chem_2026_74_25_20015_20027_Figure_5_Linear_regression_analysis_comparing_total_ergot_alkaloid_concentrations_determined_by_ELISA_and_UHPLC_MS_MS_5d7b5c5c61.jpeg" alt="J. Agric. Food Chem. 2026, 74, 25, 20015–20027: Figure 5. Linear regression analysis comparing total ergot alkaloid concentrations determined by ELISA and UHPLC-MS/MS for wheat and durum wheat cargo samples (n = 102). Each point represents the mean of duplicate measurements (extracted from the same test portion) ELISA determinations for an individual cargo sample, plotted against the corresponding UHPLC-MS/MS result. Statistical analysis was performed using linear regression." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_J_Agric_Food_Chem_2026_74_25_20015_20027_Figure_5_Linear_regression_analysis_comparing_total_ergot_alkaloid_concentrations_determined_by_ELISA_and_UHPLC_MS_MS_5d7b5c5c61.jpeg 197w," sizes="100vw" width="992" height="784">Bland-Altman analysis confirmed this proportional bias, showing a consistent negative mean difference between ELISA and UHPLC-MS/MS results across the concentration range (<a target="_blank" rel="noopener noreferrer" href="https://pubs.acs.org/doi/10.1021/acs.jafc.6c05234#fig6">Figure 6</a>). Importantly, the magnitude of disagreement did not increase randomly with concentration; rather, it reflected a systematic offset, indicating that the discrepancy was not driven by analytical imprecision or outliers but by fundamental differences in method response characteristics. This pattern indicates method-level bias rather than random analytical variability, which may arise from combined workflow effects including extraction, matrix response, and immunochemical cross-reactivity. The magnitude of the ELISA-UHPLC-MS/MS bias was substantially greater than the intermediate precision observed for the ELISA quality-control material, supporting the interpretation that the disagreement reflects systematic differences in method response rather than insufficient ELISA repeatability. Such behavior is well documented when immunoassays are compared with chromatographic methods for complex mycotoxin mixtures, including ergot alkaloids and other structurally diverse mycotoxins in cereals and feed. <a href="javascript:void(0);">(28,30,33)</a> These studies consistently report matrix- and profile-dependent bias in ELISA relative to LC-MS/MS, arising from differential antibody cross-reactivity and the summed immunoreactivity response of the assay, because ELISA produces an immunoreactivity-weighted total that depends on antibody affinity toward individual alkaloids and their relative abundance in the sample, whereas UHPLC-MS/MS quantifies compound-specific concentrations of each alkaloid and its epimers using compound-specific transitions and response factors, and total ergot alkaloid concentration is calculated as the sum of these targeted analytes. In contrast, ELISA produces an immunoreactivity-weighted total, determined by antibody affinity toward individual alkaloids and their competitive binding behavior within the assay format. Consequently, agreement between ELISA and UHPLC-MS/MS depends not only on total concentration, but also on the relative distribution of individual alkaloids within each sample. Although differences in extraction solvent composition and pH may influence ergot alkaloid recovery, stability, and epimeric distribution, each method was applied according to its intended analytical workflow. The ELISA was performed using the manufacturer-recommended acidic methanol/water extraction, whereas UHPLC-MS/MS was performed using the validated acetonitrile/aqueous ammonium carbonate extraction procedure. Therefore, the comparison reflects the performance of the complete methods as used in practice, rather than an isolated comparison of extraction solvents. The sequential extraction experiment further showed that repeating the ELISA extraction under the same kit conditions did not substantially increase the ELISA-measured response, supporting the adequacy of the kit extraction within the ELISA workflow while recognizing that it does not directly compare acidic and ammonium carbonate-based extraction conditions. Previous ELISA-LC-MS/MS comparison studies for ergot alkaloids have reported both under- and overestimation depending on assay design and matrix, including differences in alkaloid profile, cereal type, and antibody cross-reactivity. Turner et al. observed substantial overestimation of ergot alkaloids by ELISA relative to HPLC in rye field samples, attributed to strong antibody cross-reactivity toward alkaloids prevalent in rye matrices. Similar profile-dependent bias has also been reported in interlaboratory and method-comparison studies, <a href="javascript:void(0);">(33)</a> demonstrating that this behavior is not unique to a single data set.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/J_Agric_Food_Chem_2026_74_25_20015_20027_Figure_6_Bland_Altman_plot_comparing_ELISA_and_UHPLC_MS_MS_measurements_of_total_ergot_alkaloids_1be40cce50.jpeg" alt="J. Agric. Food Chem. 2026, 74, 25, 20015–20027: Figure 6. Bland-Altman plot comparing ELISA and UHPLC-MS/MS measurements of total ergot alkaloids for wheat and durum wheat cargo samples (n = 102)." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_J_Agric_Food_Chem_2026_74_25_20015_20027_Figure_6_Bland_Altman_plot_comparing_ELISA_and_UHPLC_MS_MS_measurements_of_total_ergot_alkaloids_1be40cce50.jpeg 230w," sizes="100vw" width="1437" height="977">This study provides a comprehensive evaluation of a commercially available ELISA for total ergot alkaloids, using wheat samples as the primary application matrix, and integrates performance metrics with mechanistic insights into method behavior. While the assay demonstrated strong repeatability, calibration stability, and robustness to procedural factors such as test portion mass and extraction conditions, systematic differences relative to UHPLC-MS/MS were consistently observed. By linking these differences to compound-specific antibody cross-reactivity and matrix-dependent signal modulation, this work clarifies that ELISA-derived results reflect an immunochemically weighted response rather than a stoichiometric total. This distinction is critical for the appropriate interpretation of ELISA data and supports its use within tiered analytical strategies as a rapid screening tool, complemented by UHPLC-MS/MS for confirmatory quantification and regulatory decision-making.</div>

Comparative Evaluation of Total Ergot Alkaloids Determination by ELISA and UHPLC-MS/MS in Wheat: Implications of Cross-Reactivity and Matrix Effects

In the week of July 6, the following webinars await you in the field of liquid phase and especially HPLC and LC/MS.<blockquote><h4><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinars">👉 DETAILS AND REGISTRATION IN THE WEBINARS SECTION</a></h4>👉 Search and filter in the largest global database with over 5 500 GC, LC, MS and spectroscopy webinars.</blockquote><h5>1. Waters Corporation: Characterizing Next-Generation Antibody Therapeutics: LC Strategies for Multispecifics, ADCs &amp; AOCs</h5><ul><li>Tu, 7.7.2026 11:00 CEST</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/6317">Registration</a></li></ul>In this webinar, we share a pragmatic characterization roadmap using LC-based methods across intact, subunit, and peptide-mapping workflows.<h5>2. Separation Science: Advances in Antibiotic Extraction from Water: An Automated SPE Approach</h5><ul><li>We, 8.7.2026 09:00 CEST</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/6319">Registration</a></li></ul>Streamline water analysis workflows and gain practical insights for modern environmental monitoring.<h5>3. SCIEX: Analysis of per- and polyfluoroalkyl substances (PFAS) in food substances using LC-MS/MS solutions</h5><ul><li>We, 8.7.2026 15:00 CEST</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/6322">Registration</a></li></ul>Master PFAS analysis in food safety. Learn how SCIEX LC-MS/MS solutions and QTRAP workflows overcome analytical challenges to ensure accurate and compliant result confirmation.<h5>4. Wiley: From liquids to solids: expand what your UV-Vis-NIR Spectrometer can do</h5><ul><li>We, 8.7.2026 16:00 CEST</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/6367">Registration</a></li></ul>Discover how the Agilent Cary UV-Vis-NIR spectrometer, together with Specac accessories, enables reliable analysis of liquids, powders, films, pellets, and highly scattering solid samples.<h5>5. Thermo Fisher Scientific: Advanced tools to maximize your sample throughput with Chromeleon Software and Vanquish UHPLC</h5><ul><li>Th, 9.7.2026 11:00 CEST</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/6326">Registration</a></li></ul>Learn how Chromeleon software and the Vanquish Duo UHPLC system streamline sample analysis, automate routine workflows, increase throughput, and reduce errors through practical demonstrations.<h5>6. Bruker: 4D-Metabolomics™ in Action: Real-World Strategies for Deep Discovery and Confident Annotation</h5><ul><li>Th, 9.7.2026 17:00 CEST</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/6328">Registration</a></li></ul>Explore how next-generation mass spectrometry technologies, ion mobility, and computational tools are helping researchers move from complex datasets to confident insight.<h5>7. Waters Corporation: Cracking the Code on Nitrogen Methods</h5><ul><li>Th, 9.7.2026 17:00 CEST</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/6327">Registration</a></li></ul>Learn the fundamentals of nitrogen analysis for wastewater testing. Explore analytical methods, QA/QC, sample handling, compliance reporting, and troubleshooting for accurate environmental monitoring.<h5>8. SelectScience: Next-generation metabolomics: From annotation to discovery at scale</h5><ul><li>Th, 9.7.2026 17:00 CEST</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/6281">Registration</a></li></ul>Explore how next-generation mass spectrometry, ion mobility, and advanced computational tools accelerate confident small-molecule annotation and discovery in metabolomics.<h5>9. SCIEX: LC-MS/HRMS Approaches for Monitoring Organic Micropollutants in Wastewater and Environmental Systems</h5><ul><li>Fr, 10.7.2026 15:00 CEST</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/6370">Registration</a></li></ul>Discover how advanced LC-MS/HRMS workflows enable trace-level detection of organic micropollutants, suspect and non-target screening, and environmental monitoring of wastewater systems.

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Waters XSelect HSS Cyano (CN) HPLC Columns

Waters XSelect HSS Cyano column is a low coverage, unendcapped, highly stable cyano phase that provides low hydrophobicity for the rapid elution of hydrophobic analytes

The XSelect HSS Cyano column is a low coverage, unendcapped, highly stable cyano phase that provides low hydrophobicity for the rapid elution of hydrophobic analytes as well as alternate selectivity to traditional alkyl chain columns. Packed in ultra-low dispersion hardware, 2.5 µm eXtended Performance [XP] columns enable exceptional separation performance, robustness and throughput for HPLC assays while enabling a seamless transition path toward UPLC adoption.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_X_Bridge_BEH_C18_HPLC_Column_e96c520ca3.jpg" alt="Waters: XSelect HSS Cyano (CN) HPLC Column" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_X_Bridge_BEH_C18_HPLC_Column_e96c520ca3.jpg 234w," sizes="100vw" width="660" height="440"><h4>Specifications</h4><ul><li>Chemistry: Cyano (CN)</li><li>Separation Mode: Reversed Phase</li><li>Particle Substrate: Silica</li><li>pH Range Min: 2 pH</li><li>pH Range Max: 8 pH</li><li>Maximum Pressure: 10000 psi (690 Bar)</li><li>Endcapped: Yes</li><li>Silanol Activity: High</li><li>Particle Shape: Spherical</li><li>Pore Size: 100 Å</li><li>System: UHPLC, UPLC</li><li>Particle Technology: HSS</li><li>USP Classification: L10</li><li>Carbon Load: 5 %</li></ul><h3>XSelect Columns</h3><h4>Achieve unmatched reproducibility with a wide range of selectivity</h4>Quickly and easily develop robust methods compatible with all modern chromatographic detection modes and transfer them from lab to lab with the XSelect Column family.Designed to help you achieve reproducible and highly selective chromatographic separations, the XSelect Column family features two base particles technologies, Charged Surface Hybrid (CSH) and High Strength Silica (HSS). With XSelect Columns, method development scientists like you can achieve a wide range of selectivities across all application areas, without compromising performance.<h4>Overview</h4><ul><li>Designed for selectivity and stability at low pH</li><li>Superior peak shape for basic compounds</li><li>Highest analyte mass loading available</li><li>Excellent batch-to-batch reproducibility</li><li>Ideal for rapid method development</li><li>CSH Column chemistries: C18, Fluoro-Phenyl (PFP), and Phenyl-Hexyl, available in 2.5 XP, 3.5, 5 μm analytical and 10 μm OBD preparative sizes</li><li>HSS Column chemistries: HSS C18, C18 SB, and T3, available in 2.5 XP, 3.5, and 5 μm particle sizes</li><li>Prolonged column lifetimes with optional VanGuard Cartridges specifically designed using XSelect stationary phases</li><li>Recommended Use: For improving selectivity, peak shape, and retentivity in methods development.</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_X_Select_Columns_ff2c4ddb9e/Waters-XSelect-Columns.jpg" alt="Waters: XSelect Columns" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_X_Select_Columns_ff2c4ddb9e/thumbnail_Waters-XSelect-Columns.jpg 242w," sizes="100vw" width="900" height="580">

Waters Nova-Pak Cyano (CN) HPLC Columns

The Nova-Pak Cyano (CN) Column with a small 4 µm particle size of the Nova-Pak bonded phase offers much higher resolution and faster chromatography

Nova-Pak HP CN columns are silica-based, cyano phases that are based on 4 µm particle technology. These columns can be used to achieve unique compound selectivity for both normal-phase and reversed-phase applications.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_Atlantis_d_C18_HPLC_Columns_d7a741df9f.jpg" alt="Waters: Nova-Pak Cyano (CN) HPLC Columns" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_Atlantis_d_C18_HPLC_Columns_d7a741df9f.jpg 234w," sizes="100vw" width="660" height="440"><h4>Specifications</h4><ul><li>Chemistry: Cyano (CN)</li><li>Separation Mode: Reversed Phase</li><li>Particle Substrate: Silica</li><li>pH Range Min: 2 pH</li><li>pH Range Max: 8 pH</li><li>Maximum Pressure: 6000 psi (415 Bar)</li><li>Endcapped: Yes</li><li>Silanol Activity: Medium</li><li>Particle Shape: Spherical</li><li>Pore Size: 60 Å</li><li>System: HPLC</li><li>USP Classification: L10</li><li>Carbon Load: 3 %</li></ul><h3>Nova-Pak Columns</h3><h4>Scale-up from analytical to prep, simplified</h4>Reducing solvent consumption while maintaining resolution for complex mixtures is important, but that’s a challenge with larger particle sizes. With the smaller particle sizes and shorter column lengths in Nova-Pak Columns, those problems are a thing of the past. The bonded phases of Nova-Pak Columns are available in 4 µm and 6 µm particle sizes, offering high resolution and faster, more efficient chromatography over columns packed with larger particle sizes.Nova-Pak Analytical Columns with 4 µm particle size packing offer a range of length options with 75, 100, 150, and 300 mm length steel columns. The semi-preparative Prep Nova-Pak HR Columns are packed with 6 µm particle size packings for unparalleled separation possibilities. The high-efficiency packing of Prep Nova-Pak HR Columns provides faster separations with less solvent for the added advantage of more concentrated fractions—reducing your preparative chromatography cost. To ensure batch-to-batch reproducibility, all Nova-Pak C18, C8, Cyano and Phenyl Columns are packed according to stringent quality control (QC) procedures in Waters cGMP manufacturing facility.<h4>Overview</h4><ul><li>Reduces solvent consumption while maintaining resolution for complex mixtures</li><li>Packed to stringent QC procedures to ensure batch-to-batch reproducibility</li><li>Chemistries include C18, C8, Cyano, and Phenyl stationary phases</li><li>Available in 4 µm analytical and 6 µm semi-preparative particle sizes</li><li>Recommended Use: For higher resolution and faster chromatography.</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_Nova_Pak_Columns_fad066ad02/Waters-Nova-Pak-Columns.jpg" alt="Waters: Nova-Pak Columns" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_Nova_Pak_Columns_fad066ad02/thumbnail_Waters-Nova-Pak-Columns.jpg 245w," sizes="100vw" width="900" height="392">

Waters ACQUITY UPLC HSS CN Column

Waters ACQUITY UPLC HSS CN chemistry is an ultra-performance general purpose propyl cyano bonded phase that provides superior peak shape for bases, increased retention, and extremely long column lifetimes at low pH.

ACQUITY UPLC HSS CN chemistry is an ultra-performance general purpose propyl cyano bonded phase that provides superior peak shape for bases, increased retention, and extremely long column lifetimes at low pH. ACQUITY HSS CN columns are fully scalable and transferable to HPLC separations.<h5>Specifications</h5><ul><li>Chemistry: Cyno (CN)</li><li>Separation Mode: Reversed Phase</li><li>Particle Substrate: Silica</li><li>pH Range Min: 2 pH</li><li>pH Range Max: 8 pH</li><li>Endcapped: No</li><li>Silanol Activity: High</li><li>Particle Shape: Spherical</li><li>Pore Size: 100 Å</li><li>System: UHPLC, UPLC</li><li>Particle Technology: HSS</li><li>USP Classification: L10</li><li>Carbon Load: 5 %</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_ACQUITY_UPLC_BEH_C8_Column_9c78ca994d.jpg" alt="Waters: ACQUITY UPLC HSS CN Column" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_ACQUITY_UPLC_BEH_C8_Column_9c78ca994d.jpg 234w," sizes="100vw" width="660" height="440"><h3>ACQUITY UPLC Columns</h3><h5>Increase speed, sensitivity, and resolution, without compromising data integrity</h5>Increasing sample throughput, achieving unmatched resolution, and improving analyte sensitivity are priorities for your separation goals. With Waters ACQUITY UPLC Columns, you can meet all your method requirements and substantially reduce your analysis time and cost-per-sample.Waters ACQUITY UPLC Columns’ sub-2-µm particle chemistries enable chromatographers to harness the separation efficiency improvements at increased flow rates of ultra-performance liquid chromatography (UPLC) technology, thereby improving analyte resolution and sensitivity as well as business productivity and profitability. &nbsp;<h4>Overview</h4><ul><li>Best-in-class column efficiency for superior separation speed, sensitivity, and resolution</li><li>8 UPLC particles of various substrate technologies, including Ethylene Bridged Hybrid (BEH), High Strength Silica (HSS), and Charged Surface Hybrid (CSH)</li><li>22 stationary phases, including C18, Phenyl-Hexyl, C8, Embedded-Polar, C4, HILIC, Amide, Diol, Cyano, and PFP</li><li>7 application-specific UPLC Column chemistries for SEC, proteins, peptides, oligonucleotides, glycan, and amino acid analysis</li><li>More than 50 different column dimensions and configurations, with internal column diameters ranging from 1.0–4.6 mm and column lengths ranging from 30–300 mm</li><li>Prolonged column lifetimes with optional VanGuard Cartridges specifically designed using ACQUITY stationary phases</li><li>Recommended Use: For improved speed, sensitivity, and resolution in methods development.</li></ul><h4>Particle technology</h4><h5>BEH - Ethylene Bridged Hybrid Particle Technology</h5>ACQUITY UPLC BEH 1.7 µm fully porous columns provide superior analyte peak shape, efficiency, and chemical stability to any separation. ACQUITY UPLC BEH Columns are available in both reversed-phase and HILIC separation modes, with chemistries that provide selectivities for a wide range of compounds. With the ruggedness to operate at extreme pH conditions, ACQUITY UPLC BEH Columns enable the separation scientist to use a wide pH range to influence retention, selectivity, and sensitivity for ionizable compounds.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_BEH_technology_c07876b8a3.jpg" alt="Waters: BEH particle technology" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_BEH_technology_c07876b8a3.jpg 170w," sizes="100vw" width="1050" height="964"><h5>CSH - Charged Surface Hybrid Particle Technology</h5>ACQUITY UPLC CSH™ 1.7 µm fully porous columns provide superior peak shape for basic analytes when using low ionic strength acidic mobile phases (e.g. 0.1% formic acid). With CSH Technology, the controlled low-level positive surface charge provides exceptional peak shape for basic analytes which can be achieved without the use of ion pairing reagents. Typically reversed-phase bonded phases have poor peak shape for basic compounds in formic acid even at analytical mass loads. Built on the rugged Ethylene Bridged Hybrid (BEH) particle, ACQUITY UPLC CSH phases can withstand the toughest operating conditions.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_CSH_technology_ff4ddc1708.jpg" alt="Waters: CSH particle technology" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_CSH_technology_ff4ddc1708.jpg 169w," sizes="100vw" width="1049" height="968"><h5>HSS - High Strength Silica (HSS) Particle Technology</h5>ACQUITY UPLC HSS 1.8 µm fully porous columns provide expected silica particle-like performance to the UPLC family. The HSS particles’ high surface area, and lower pore volume provide increased retentivity over the hybrid particles. This robust particle technology is available in five uniquely different bonded phases, which truly maximize the selectivity space. The wide array of bonded phases associated with ACQUITY UPLC HSS Columns enable traditional hydrophobic reversed-phase interactions along with dipole-dipole interactions, aromatic interactions, and hydrogen bonding interactions.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_HSS_technology_b12d3ddfab.jpg" alt="Waters: HSS particle technology" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_HSS_technology_b12d3ddfab.jpg 162w," sizes="100vw" width="975" height="936"><h4>Get superior peak shape for basic analytes</h4>Reversed-phase bonded phases typically have poor peak shape for basic compounds when using formic acid, even at analytical mass loads. However, ACQUITY UPLC CSH Columns are the exception. These rugged UPLC Columns provide superior peak shape for basic analytes with formic acid or other low ionic-strength, acidic mobile phases. The controlled, low-level, positive surface charge bonded to the ethylene-bridged hybrid (BEH) particles provides excellent peak shape for bases—without the need for the use of ion-pairing reagents.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_Analysis_of_ziprasidone_peroxide_degradation_on_an_ACQUITY_UPLC_CSH_C18_Column_56909f1dc8.jpeg" alt="Waters: Analysis of ziprasidone peroxide degradation on an ACQUITY UPLC CSH C18 Column" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_Analysis_of_ziprasidone_peroxide_degradation_on_an_ACQUITY_UPLC_CSH_C18_Column_56909f1dc8.jpeg 245w," sizes="100vw" width="1024" height="485"><h4>The ruggedness you want, the repeatability you need</h4>ACQUITY UPLC BEH 1.7 µm fully porous columns provide superior analyte peak shape, efficiency, and chemical stability to any separation. ACQUITY UPLC BEH Columns are available in both reversed-phase and HILIC separation modes, with chemistries that provide selectivities for a wide range of compounds. With the ruggedness to operate at extreme pH conditions, ACQUITY UPLC BEH Columns enable separation scientists to use a wide pH range to influence retention, selectivity, and sensitivity for ionizable compounds.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_ACQUITY_UPLC_BEH_Columns_can_be_used_at_the_extremes_of_mobile_phase_p_H_1_12_to_enhance_retention_and_specificity_for_complex_samples_that_include_mixtures_of_acidic_basic_and_neutral_analytes_5104d9a155.jpeg" alt="Waters: ACQUITY UPLC BEH Columns can be used at the extremes of mobile phase pH (1-12) to enhance retention and specificity for complex samples that include mixtures of acidic, basic, and neutral analytes." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_ACQUITY_UPLC_BEH_Columns_can_be_used_at_the_extremes_of_mobile_phase_p_H_1_12_to_enhance_retention_and_specificity_for_complex_samples_that_include_mixtures_of_acidic_basic_and_neutral_analytes_5104d9a155.jpeg 195w," sizes="100vw" width="1024" height="823"><h4>Achieve flexibility in method development</h4>Chromatographic laboratories need to quickly and easily develop robust methods that are compatible with all modern chromatographic detection modes and transferrable to laboratories and sites that operate different LC system platforms. With ACQUITY UPLC CSH and XSelect Columns, method development scientists in all application areas can achieve a wide range of selectivities without compromising performance across attributes, including superior peak shape for basic compounds, low column bleeds, excellent batch-to-batch reproducibility, and high efficiency.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_CSH_Columns_offer_a_wide_range_of_selectivity_differences_large_S_values_733305abf8.jpeg" alt="Waters: CSH Columns offer a wide range of selectivity differences (large S-values)." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_CSH_Columns_offer_a_wide_range_of_selectivity_differences_large_S_values_733305abf8.jpeg 136w," sizes="100vw" width="1024" height="1173">

Products from category CN HPLC columns focused on Consumables distributed by Waters Corporation (Europe) - page 1

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